# Patent

## Title
N-[4-Fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide as an HIV integrase inhibitor

## Abstract

The invention encompasses a series of bicyclic pyrimidinone compounds of Formula I which inhibit HIV integrase and prevent viral integration into human DNA. This action makes the compounds useful for treating HIV infection and AIDS. The invention also encompasses pharmaceutical compositions and methods for treating those infected with HIV.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation-in-part of U.S. application Ser. No. 11/288,533 filed on Nov. 29, 2005 now U.S. Pat. No. 7,157,447 which is a continuation-in-part of U.S. Nonprovisional application Ser. No. 11/126,891 filed May 11, 2005 now U.S. Pat. No. 7,176,196 and claims the benefit of U.S. Provisional Application Ser. Nos. 60/603,371 filed Aug. 20, 2004 and 60/575,513 filed May 28, 2004.


BACKGROUND OF THE INVENTION
Human immunodeficiency virus (HIV) has been identified as the etiological agent responsible for acquired immune deficiency syndrome (AIDS), a fatal disease characterized by destruction of the immune system and the inability to fight off life threatening opportunistic infections. Recent statistics (UNAIDS: Report on the Global HIV/AIDS Epidemic, December 1998), indicate that as many as 33 million people worldwide are infected with the virus. In addition to the large number of individuals already infected, the virus continues to spread. Estimates from 1998 point to close to 6 million new infections in that year alone. In the same year there were approximately 2.5 million deaths associated with HIV and AIDS.
There are currently a number of antiviral drugs available to combat the infection. These drugs can be divided into three classes based on the viral protein they target and their mode of action. In particular, saquinavir, indinavir, ritonavir, nelfinavir and amprenavir are competitive inhibitors of the aspartyl protease expressed by HIV. Zidovudine, didanosine, stavudine, lamivudine, zalcitabine and abacavir are nucleoside reverse transcriptase inhibitors that behave as substrate mimics to halt viral cDNA synthesis. The non-nucleoside reverse transcriptase inhibitors, nevaripine, delavirdine and efavirenz inhibit the synthesis of viral cDNA via a non-competitive (or uncompetitive) mechanism. Used alone these drugs are effective in reducing viral replication. The effect is only temporary as the virus readily develops resistance to all known agents. However, combination therapy has proven very effective at both reducing virus and suppressing the emergence of resistance in a number of patients. In the US, where combination therapy is widely available, the number of HIV-related deaths has declined (Palella, F. J.; Delany, K. M.; Moorman, A. C.; Loveless, M. O.; Further, J.; Satten, G. A.; Aschman, D. J.; Holmberg, S. D. N. Engl. J. Med. 1998, 338, 853-860).
Unfortunately, not all patients are responsive and a large number fail this therapy. In fact, approximately 30-50% of patients ultimately fail combination therapy. Treatment failure in most cases is caused by the emergence of viral resistance. Viral resistance in turn is caused by the rapid turnover of HIV-1 during the course of infection combined with a high viral mutation rate. Under these circumstances incomplete viral suppression caused by insufficient drug potency, poor compliance to the complicated drug regiment as well as intrinsic pharmacological barriers to exposure provides fertile ground for resistance to emerge. More disturbing are recent findings which suggest that low-level replication continues even when viral plasma levels have dropped below detectable levels (<50 copies/ml) (Carpenter, C. C.; Cooper, D. A.; Fischl, M. A.; Gatell, J. M.; Gazzard, B. G.; Hammer, S. M.; Hirsch, M. S.; Jacobsen, D. M.; Katzenstein, D. A.; Montaner, J. S.; Richman, D. D.; Saag, M. S.; Schechter, M.; Schooley, R. T.; Thompson, M. A.; Vella, S.; Yeni, P. G.; Volberding, P. A. JAMA 2000, 283, 381-390). Clearly there is a need for new antiviral agents, preferably targeting other viral enzymes to reduce the rate of resistance and suppress viral replication even further.
HIV expresses three enzymes, reverse transcriptase, an aspartyl protease, and integrase. All three are targets for treating AIDS and HIV infection. HIV integrase catalyzes insertion of the viral cDNA into the host cell genome, which is a critical step in the viral life cycle. HIV integrase inhibitors belonging to a class of diketo acid compounds prevented viral integration and inhibited HIV-1 replication in cells (Hazuda et al. Science 2000, 287, 646). And recently, HIV integrase inhibitors have been accepted into clinical trials for treating AIDS and HIV infection (Neamati Expert. Opin. Ther. Patents 2002, 12, 709, Pais and Burke Drugs Fut. 2002, 27, 1101).



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is an observed and simulated powder X-ray diffraction pattern of form N-1 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.
FIG. 2 is a differential scanning calorimetry thermogram of form N-1 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.
FIG. 3 is a thermal gravimetric analysis scan of form N-1 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.
FIG. 4 is a SSNMR spectra of form N-1 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.
FIG. 5 is a moisture sorption isotherm of form N-1 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.
FIG. 6 is an observed and simulated powder X-ray diffraction pattern of form N-2 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.
FIG. 7 is a differential scanning calorimetry thermogram of form N-2 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.
FIG. 8 is a thermal gravimetric analysis scan of form N-2 of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide.



DESCRIPTION OF THE INVENTION
The invention encompasses compounds of Formula I, including pharmaceutically acceptable salts and solvates, their pharmaceutical compositions, and their use in inhibiting HIV integrase and treating those infected with HIV or AIDS.
One aspect of the invention are compounds of Formula I




wherein:

R1 is C1-6(Ar1)alkyl, C1-6(Ar1)(CON(R8)(R9))alkyl, C1-6(Ar1)(CO2R14)alkyl, C1-6(Ar1)hydroxyalkyl, or C1-6(Ar1)oxyalkyl;

R2 is hydrogen, C1-6alkyl, or OR14;

R3 is hydrogen, halo, hydroxy, cyano, C1-6alkyl, C3-7cycloalkyl, C5-7cycloalkenyl, C1-6haloalkyl, C1-6alkoxy, C1-6alkylthio, C1-6haloalkoxy, N(R8)(R9), NHAr2, N(R6)SO2R7, N(R6)COR7, N(R6)CO2R7, OCOR7, OCO2R7, OCON(R8)(R9), OCH2CO2R7, OCH2CON(R8)(R9), COR6, CO2R6, CON(R8)(R9), SOR7, S(═N)R7, SO2R7, SO2N(R6)(R6), PO(OR6)2, C24(R12)alkynyl, R13, Ar2, or Ar3;

R4 is hydrogen, halo, hydroxy, cyano, C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy, or N(R6)(R6);

R5 is hydrogen, halo, hydroxy, cyano, C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy, or N(R6)(R6);

R6 is hydrogen, C1-6alkyl, or C3-7cycloalkyl;

R7 is C1-6alkyl or C3-7cycloalkyl;

R8 is hydrogen, C1-6alkyl, C1-6hydroxyalkyl, C1-6(C1-6alkoxy)alkyl or C1-6(C1-6dialkylamino)alkyl;

R9 is hydrogen, C1-6alkyl, C1-6hydroxyalkyl, C1-6(C1-6alkoxy)alkyl or C1-6(C1-6dialkylamino)alkyl; or

N(R8)(R9) taken together is azetidinyl, pyrrolidinyl, (R10)-piperidinyl, N—(R11)-piperazinyl, morpholinyl, thiomorpholinyl, or dioxothiazinyl;

R10 is hydrogen, C1-6alkyl, or C1-6hydroxyalkyl;

R11 is hydrogen, C1-6alkyl, C3-7cyclolkyl, COR6, or CO2R6;

R12 is hydrogen, hydroxy, N(R6)(R6), SO2R7, OSO2R7, or dioxothiazinyl;

R13 is azetidinonyl, pyrrolidinonyl, valerolactamyl, caprolactamyl, maleimido, oxazolidinyl, or dioxothiazinyl, and is substituted with 0-1 substituents selected from the group consisting of hydroxymethyl, acetoxymethyl, and aminomethyl;

R14 is hydrogen or C1-6alkyl;





Ar2 is tetrazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, furanyl, thienyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyridinyl, hydroxypyridinyl, quinolinyl, isoquinolinyl, or indolyl, and is substituted with 0-2 substituents selected from the group consisting of halo, cyano, benzyl, C1-6alkyl, C1-6alkoxy, N(R8)(R9), CON(R8)(R9), CO2R6, CONHSO2N(R6)(R6), CONHSO2N(R6)(phenyl), and CONHSO2N(R6)(halophenyl);

Ar3 is phenyl substituted with 0-2 substituents selected from the group consisting of halo, cyano, hydroxy, C1-6alkyl, C1-6alkoxy, (C1-6alkoxy)methyl, C1-6haloalkyl, C1-6haloalkoxy, N(R8)(R9), CON(R6)(R6), and CH2N(R8)(R9), or is dioxolanylphenyl; and

X—Y—Z is C(R14)2OC(R14)2, C(R14)2OC(R14)2C(R14)2, or C(R14)2OC(R14)2C(R14)2C(R14)2;

or a pharmaceutically acceptable salt or solvate thereof.

Another aspect of the invention is a compound of Formula I where R1 is C1-6(Ar1)alkyl.
Another aspect of the invention is a compound of Formula I where R1 is




Another aspect of the invention is a compound of Formula I where R1 is




Another aspect of the invention is a compound of Formula I where R2 is hydrogen.
Another aspect of the invention is a compound of Formula I where R3 is hydrogen, halo, N(R8)(R9), N(R6)COR7, OCON(R8)(R9), CON(R8)(R9), SOR7, SO2R7, SO2N(R6)(R6), PO(OR6)2, R13, or Ar2.
Another aspect of the invention is a compound of Formula I where X—Y—Z is C(R14)2OCH2, C(R14)2OCH2CH2, or C(R14)2OCH2CH2CH2.
Another aspect of the invention is a compound of Formula I where X—Y—Z is CH2OCH2, C(CH3)HOCH2, C(CH3)2OCH2, CH2OCH2CH2, C(CH3)HOCH2CH2, C(CH3)2OCH2CH2, CH2OCH2CH2CH2, C(CH3)HOCH2CH2CH2, or C(CH3)2OCH2CH2CH2.
Another aspect of the invention is a compound of Formula I selected from the group of structures consisting of




Another aspect of the invention is a method for making a compound of formula II




where:

R1 is C1-6(Ar1)alkyl;

R2 is hydrogen;

R3 is hydrogen, halo, hydroxy, cyano, C1-6alkyl, C3-7cycloalkyl, C5-7cycloalkenyl, C1-6haloalkyl, C1-6alkoxy, C1-6alkylthio, C1-6haloalkoxy, N(R8)(R9), NHAr2, N(R6)SO2R7, N(R6)COR7, N(R6)CO2R7, OCOR7, OCO2R7, OCON(R8)(R9), OCH2CO2R7, OCH2CON(R8)(R9), COR6, CO2R6, CON(R8)(R9), SOR7, S(═N)R7, SO2R7, SO2N(R6)(R6), PO(OR6)2, C24(R12)alkynyl, R13, Ar2, or Ar3;

R4 is hydrogen, halo, hydroxy, cyano, C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy, or N(R6)(R6);

R5 is hydrogen, halo, hydroxy, cyano, C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy, or N(R6)(R6);

R6 is hydrogen, C1-6alkyl, or C3-7cycloalkyl;

R7 is C1-6alkyl or C3-7cycloalkyl;

R8 is hydrogen, C1-6alkyl, C1-6hydroxyalkyl, C1-6(C1-6alkoxy)alkyl or C1-6(C1-6dialkylamino)alkyl;

R9 is hydrogen, C1-6alkyl, C1-6hydroxyalkyl, C1-6(C1-6alkoxy)alkyl or C1-6(C1-6dialkylamino)alkyl; or

N(R8)(R9) taken together is azetidinyl, pyrrolidinyl, (R10)-piperidinyl, N—(R11)-piperazinyl, morpholinyl, thiomorpholinyl, or dioxothiazinyl;

R10 is hydrogen, C1-6alkyl, or C1-6hydroxyalkyl;

R11 is hydrogen, C1-6alkyl, C3-7cyclolkyl, COR6, or CO2R6;

R12 is hydrogen, hydroxy, N(R6)(R6), SO2R7, OSO2R7, or dioxothiazinyl;

R13 is azetidinonyl, pyrrolidinonyl, valerolactamyl, caprolactamyl, maleimido, oxazolidinyl, or dioxothiazinyl, and is substituted with 0-1 substituents selected from the group consisting of hydroxymethyl, acetoxymethyl, and aminomethyl;

R14 is hydrogen or C1-6alkyl;





Ar1 is





Ar2 is tetrazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, furanyl, thienyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyridinyl, hydroxypyridinyl, quinolinyl, isoquinolinyl, or indolyl, and is substituted with 0-2 substituents selected from the group consisting of halo, cyano, benzyl, C1-6alkyl, C1-6alkoxy, N(R8)(R9), CON(R8)(R9), CO2R6, CONHSO2N(R6)(R6), CONHSO2N(R6)(phenyl), and CONHSO2N(R6)(halophenyl); and

Ar3 is phenyl substituted with 0-2 substituents selected from the group consisting of halo, cyano, hydroxy, C1-6alkyl, C1-6alkoxy, (C1-6alkoxy)methyl, C1-6haloalkyl, C1-6haloalkoxy, N(R8)(R9), CON(R6)(R6), and CH2N(R8)(R9), or is dioxolanylphenyl;

comprising reacting a compound of the structure





where R15 is C1-6alkyl with a compound of the formula C1-6(Ar1)alkylamine under basic conditions.

Another aspect of the invention is the method for making a compound of Formula II where the base is a C1-4trialkylamine, C1-4dialkylamine, C1-4alkylamine, or ammonia.
Another aspect of the invention is the method for making a compound of Formula II where the base is a C1-4-trialkylamine or C1-4dialkylamine
Another aspect of the invention is the method for making a compound of Formula II where the base is a C1-4trialkylamine.
Another aspect of the invention the method for making a compound of Formula II where the base is triethylamine or diisopropylethylamine.
Another aspect of the invention is the method for making a compound of Formula II where the conditions include a solvent selected from the group consisting of a C1-4alcohol, a (C1-4)dialkyl ether, tetrahydrofuran, dioxane, a (C1-4)-((C1-4)dialkoxy)alkane, DMF, DMSO, or methylene chloride.
Another aspect of the invention is the method for making a compound of Formula II further comprising heating a compound of the structure




in a solvent to form a compound of the structure





Another aspect of the invention is a method for making a compound of Formula II comprising reacting a compound of the structure




where R15 is hydrogen with a compound of the formula C1-6(Ar1)alkylamine under peptide coupling conditions.

Another aspect of the invention is a method for making a compound of Formula II comprising transforming a compound of the structure




where R15 is hydrogen into the corresponding acyl chloride and reacting the acyl chloride with a compound of the formula C1-6(Ar1)alkylamine under basic conditions.

Another aspect of the invention is a compound of the structure




where R14 is hydrogen or C1-6alkyl and R15 is C1-6alkyl.

Another aspect of the invention is a compound of the structure




where R14 is methyl and R15 is methyl or ethyl.

For a compound of Formula I, any scope of R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15, Ar1, Ar2, Ar3, and X—Y—Z can be used independently with any scope of any other substituent.
“Alkyl,” “alkoxy,” “hydroxyalkyl,” and related terms with an alkyl moiety include straight and branched configurations. A term such as “C1-6(R)alkyl” means a straight or branched alkyl group of one to six carbons substituted with the substituent R. “Haloalkyl” and “halophenyl” include all permutations of halogenated alkyl or phenyl groups, from monohalo to perhalo. “Aryl” means an aromatic ring system and includes carbocyclic and heterocyclic systems. Some substituents are divalent, such as X—Y—Z. Asymmetric divalent substituents may be attached in either of the two configurations.
“C1-6(Ar1)oxyalkyl” means Ar1 is attached at the oxygen.
“Dioxolanyphenyl” means




“Dioxothiazinyl” means





The invention includes all pharmaceutically acceptable salt forms of the compounds. Pharmaceutically acceptable salts are those in which the counter ions do not contribute significantly to the physiological activity or toxicity of the compounds and as such function as pharmacological equivalents. These salts can be made according to common organic techniques employing commercially available reagents. Some anionic salt forms include acetate, acistrate, besylate, bromide, chloride, citrate, fumarate, glucouronate, hydrobromide, hydrochloride, hydroiodide, iodide, lactate, maleate, mesylate, nitrate, pamoate, phosphate, succinate, sulfate, tartrate, tosylate, and xinofoate. Some cationic salt forms include ammonium, aluminum, benzathine, bismuth, calcium, choline, diethylamine, diethanolamine, lithium, magnesium, meglumine, 4-phenylcyclohexylamine, piperazine, potassium, sodium, tromethamine, and zinc.
The invention also includes all solvated forms of the compounds, particularly hydrates. Solvates do not contribute significantly to the physiological activity or toxicity of the compounds and as such function as pharmacological equivalents. Solvates may form in stoichiometric amounts or may form from adventitious solvent or a combination of both. One type of solvate is hydrate, and some hydrated forms include monohydrate, hemihydrate, and dihydrate.
Some of the compounds of the invention exist in stereoisomeric forms. The invention includes all stereoisomeric forms of the compounds including enantiomers and diastereomers. An example of enantiomers is shown below. Methods of making and separating stereoisomers are known in the art.




The invention includes all tautomeric forms of the compounds. An example of a tautomeric pair is shown below.




Synthetic Methods
The compounds of this invention can be made by various methods known in the art including those of the following schemes and in the specific embodiments section. The variables shown in the synthetic schemes are distinct from and should not be confused with the variables in the claims or the rest of the specification. The variables in the schemes are meant only to illustrate how to make some of the compounds of this invention.
Some compounds can be synthesized from an appropriately substituted heterocycle I-1 according to Scheme I, where Ra and P can serve as protecting groups (see Greene, T. W. and Wutz, P. G. M. Protective Groups in Organic Synthesis, Second Edition, 1991, John Wiley and Sons, New York). When P is benzyl or substituted benzyl it can be removed by hydrogenolysis (H2-Pd/C) or acid hydrolysis (trifluoroacetic acid) to yield intermediate I-2. I-2 can be transaminated to I-4 by reaction with amine I-3. In a number of cases this reaction can be carried out by heating I-3 and I-2 together in the presence of base. Alternatively, standard amide coupling reagents can be used to effect the formation of the amide bond. When Ra is a lower alkyl group, Ra can be removed under ester hydrolysis conditions, such as treatment with NaOH, LiOH, or KOH to deliver the corresponding carboxylic acid I-5. Alternatively, Ra can be removed by nucleophilic displacement using NaI. When Ra is benzyl and substituted benzyl, Ra can be removed by hydrogenolysis. Intermediate I-5 can be coupled using amide bond forming reagents such as BOP, DCC, EDCI, PyBrop, PyBop or other reagents (see March, J. Advanced Organic Chemistry, Fourth Edition 1992 John Wiley & Sons, New York). The resulting intermediate I-6 can be deprotected as described for intermediate I-1.




In Scheme II, intermediate II-3 can be prepared using methods similar to those described in Sunderland, J. S.; Botta, M.; Aime, S.; Raymond, K. N. Inorg. Chem. (2001), 40, 6756-6756, where II-1 and II-2 are condensed, to provide intermediate II-3. This reaction is usually conducted in the presence of a base such as sodium hydride (NaH), sodium ethoxide (EtONa) or lithium hexamethyldisilazide (LiHMDS). Using the methods described in the reference, II-3 can be condensed with an appropriately substituted amidine II-4 to form II-5. Substituent B can be a leaving group, such as -halo (Cl, Br or I) or can be converted to a leaving group under appropriate conditions such as by forming the corresponding methylsulfonate ester. When substituent B is a methyl sulphide group it can be treated with iodomethane to form a dimethylsulfonium intermediate which is activated towards nucleophilic attack to effect ring closure.




In Scheme III, intermediate II-3 can be condensed with a cyclic-amidine to yield intermediate I-1. Intermediate III-1 can be prepared using known methods (see Patai, S, and Rappoport, Z. The Chemistry of Amidines and Imidates, Volume 2, 1991, John Wiley & Sons, New York).




In Scheme IV, nitrile IV-1, possessing a potential leaving group B, can be reacted with hydroxylamine to form intermediate IV-2. This intermediate can be reacted with a suitably protected alkyne to form IV-3 which can rearrange to from intermediate IV-4 according to literature methods (Culbertson, T. P. Journal of Heterocyclic Chemistry, 1979, 16, 1423-1424).




As shown in Scheme V, 2-(methylthio)ethanol can be alkylated with an appropriate α-haloacetic acid (V-1) wherein X is a leaving group such as Cl, Br, OTs, OMs or OTf, to deliver intermediate V-2. Following this, the carboxylic acid can be transformed to the corresponding amidine derivative using known synthetic methods (Geilen et al. Tetrahedron Letters 2002, 43, 419-421). As described in the above, the amidine can further be reacted with intermediate V-5, in the presence of a base (for example, sodium ethoxide) affording intermediate V-6. Methylation of the sulphide ether can be accomplished by treating V-6 with iodomethane and the resulting sulfonium derivative (V-7) treated with base to form the bicyclic template V-8. This intermediate can be used in the synthesis of final compounds using methods described in Scheme I.




In Scheme VI, 3-methylthiopropanal is converted to dioxolane VI-1 using well known chemistry. Treatment with trimethylsilylcyanide (TMSCN), in the presence of zinc iodide (ZnI2) produces intermediate VI-2. Reaction with ammonia provides amidine VI-3 which is used in the synthesis of pyrimidinone VI-4 according to the methods described in the previous schemes. Subsequent treatment with CH3SO2Cl and triethylamine (Et3N) results in the corresponding bicyclic intermediate VI-5. Completion of the synthesis can be carried out as illustrated in Scheme I.




Another method is illustrated in Scheme VII. This synthetic path begins with an appropriately substituted ketone which can be transformed to the corresponding nitrile intermediate VII-1. This in turn can be reacted with 2-chloroethanol to produce compound VII-2, which can be reacted with hydroxylamine and an acetylene dicarboxylate ester to yield intermediate VII-4. Heating of the intermediate can yield intermediate VII-5. Synthesis of the corresponding amide derivatives can be accomplished according to Scheme I.




In Scheme VIII, benzylation of the hydroxyl group of VII-5, as a means of functional group protection, can be achieved using benzyl bromide under basic conditions (for example, K2CO3 or NaH). Saponification of the ester group of VIII-1 can provide VIII-2 which can be coupled with appropriately substituted amines (R1R2NH) using well known amide bond forming reagents, such as benzotriazole-1-yloxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP) or O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU). Alternatively, the corresponding acid chloride can be formed, by treatment with oxalyl chloride, and reacted with an appropriate amine to form the amide bond. Removal of the benzyl group can be accomplished under a variety of conditions including treatment with CF3CO2H or H2 (Pd—C).




In yet another method, some compounds of this invention can be synthesized according to Scheme IX. In Scheme IX, pyrimidinone IX-3, can be produced using methods similar to those described in the previous schemes. This intermediate can be carried on to the final product according to a variety of paths. In one, the hydroxyl group can be benzoylated to provide intermediate IX-4 which can be further treated with K2CO3 to effect ring closure to form the bicyclic template IX-5. Alternatively, direct treatment of IX-3 with K2CO3 can provide intermediate IX-6. Intermediates IX-5, an IX-6 can be used in the synthesis the final products using the methods described in Scheme I.




In Scheme X, IX-3 can be used to synthesize the benzylated intermediate X-1. This intermediate can be carried on to final product using methods analogous to those described in Scheme VIII.




The synthesis of (2-(aminomethyl)-5-fluorophenyl)(morpholino)methanone hydrochloride is illustrated in Scheme XI.




In Scheme XII, bicyclic intermediate XII-1, prepared according to the methods described above, can be saponified using well known methods. The resulting carboxylic acid, XII-3, can then be coupled to amine XII-2 using standard amide bond forming reagents and methods. Removal of the benzyl group, by hydrogenolysis or acid mediated hydrolysis provides the final products.




An alternative route to compounds similar to those presented in Scheme XII is given in Scheme XIII. In this scheme the ester group of XIII-1 can be hydrolyzed and the resulting carboxylic acid coupled to methyl 2-(aminomethyl)-5-fluorobenzoate. A second hydrolysis reaction can produce XIII-4 which can be coupled with a second amine. This is followed by removal of the benzyl group to provide the final products.




In yet another method, Scheme XIV illustrates the synthesis of sulfonamide containing examples, starting from 5-fluoro-2-methylbenzene-1-sulfonlyl chloride.




Further illustration of methods used for the synthesis of certain compounds of the invention is shown in Scheme XV. Methylation of 5-(2-bromo-5-fluorophenyl)-1H-tetrazole can yield a mixture of XV-1 and XV-2 that can be separated and each of the compounds carried on to the corresponding final products.




Some diazarine and diaziradine analogues can be synthesized according to Scheme XVI.




Some examples of the invention can be synthesized according to the methods illustrated in Schemes XVII-XX.
















In Scheme XXI, 2-bromo-benzonitrile or 2-bromo-4-fluoro-benzonitrile can be coupled with an appropriate amide to provide XXI-1. Reduction of the nitrile group provides XXI-2 which can be used in the synthesis of the final product according to the methods described in the previous schemes. In most cases, compound XXI-1 need not be isolated but can be carried directly into the coupling reaction to form XXI-3.




The examples and methods shown in Scheme XXII are similar to those depicted in Scheme XXI.




Schemes XXIV and XXV further illustrate methods useful for the synthesis of some compounds of the invention.








In Scheme XXVI, intermediate XXVI-1 can be used to synthesize intermediates XXVI-2 via palladium catalyzed coupling. These intermediates can be further modified to provide some of the compounds of this invention.




Another method is illustrated in Scheme XXVII.




In Scheme XXVIII, compound XXVIII-1 can be converted to the corresponding methylsulfonate derivate, XXVIII-2, which can be subsequently treated with sodium azide to yield XXVIII-3. This compound in turn can serve as an intermediate for further transformation as illustrated in the scheme.




Certain examples of the current invention can be synthesized according to the methods illustrated in Schemes XXIX-XXXVIII.








































Biological Methods
HIV-Integrase Inhibition Activity. To evaluate in-vitro activity against HIV-integrase, 5 pmole of biotin labeled substrate DNA was bound to 100 μg of Streptavidin coated PVT SPA beads (Amersham Pharmacia Biotech). Recombinant integrase (0.26 ng) was incubated with the beads for 90 min at 37° C. Unbound enzyme was removed by washing the complex followed by addition of inhibitors and 0.1 fmol of P33 labeled target DNA. The reaction was stopped by adding EDTA to a final concentration of 10 mM. Samples were counted in TopCountNXT (Packard) and the CPM was used as a measure of integration. The reaction condition was as described in A. Engelman and R. Craigie, J. Virol 69, 5908-5911 (1995). The sequences of substrate and target DNA were described in Nucleic Acid Research 22, 1121-1122 (1994). Results are shown in the Table 1. Activity equal to A refers to a compound having IC50=0.002 to 0.10 μM while B and C denote compounds having IC50=0.1 to 1.0 μM and IC50≧1.0 μM respectively.










TABLE 1







Example
Activity



















1
A



2
A



3
A



4
A



5
A



6
A



7
A



8
A



9
A



10
A



11
A



12
A



13
A



14
A



15
A



16
A



17
A



18
A



19
A



20
A



21
A



22
B



23
A



24
B



25
B



26
A



27
A



28
A



29
A



30
A



31
A



32
A



33
A



34
A



35
A



36
A



37
A



38
A



39
A



40
A



41
A



42
A



43
A



44
A



45
A



46
A



47
A



48
A



49
A



50
A



51
A



52
A



53
A



54
A



55
A



56
A



57
A



58
A



59
A



60
A



61
A



62
A



63
A



64
A



65
A



66
A



67
A



68
A



69
A



70
A



71
A



72
A



73
A



74
A



75
A



76
A



77
A



78
A



79
A



80
A



81
A



82
A



83
C



84
A



85
A



86
A



87
A



88
A



89
A



90
A



91
A



92
A



93
A



94
A



95
A



96
A



97
A



98
A



99
A



100
A



101
A



102
A



103
A



104
B



105
A



106
A



107
A



108
A



109
A



110
A



111
A



112
A



113
A



114
A



115
A



116
A



117
A



118
A



119
A



120
A



121
A



122
A



123
A



124
A



125
A



126
A



127
A



128
A



129
A



130
A



131
B



132
A



133
B



134
A



135
A



136
A



137
A



138
A



139
A



140
A



141
A



142
A



143
A



144
B



145
C



146
A



147
A



148
B



149
A



150
A



151
A



152
A



153
A



154
A



155
A



156
A



157
A



158
A



159
B



160
A



161
A



162
A



163
A



164
A



165
A



166
B



167
A



168
A



169
A



170
A



171
A



172
A



173
A



174
A



175
A



176
A



177
A



178
A



179
A



180
A



181
A



187
A



188
A



189
A



190
A



191
A



192
A



193
A



194
A



195
A



196
A



197
A



198
A



199
A



200
A



201
A



202
A



203
A



204
A



205
A



206
A



207
A



208
A



209
A



210
A



211
A



212
A



213
A



214
C



215
A



216
A



217
A



218
A



219
A



220
A



221
A



222
A



223
A



224
A



225
A



226
A



227
A



228
A



229
A



230
A



231
A



232
A



233
A



235
A



236
A



237
A



238
A



239
A



240
A



241
A



242
A



243
A



244
A



245
A



246
A



247
A



248
A



249
A



250
A



251
A



252
B



253
C



254
A



255
A



256
B



257
A



258
A



259
A



260
A



261
A



262
A



263
A



264
A



265
A



266
A



267
A



268
A



269
A



270
A



271
A



272
A



273
A



274
A



275
A



276
A



277
A



278
A



279
A



280
A



281
A



282
A










Inhibition of HIV replication. A recombinant NL-Rluc virus was constructed in which a section of the nef gene from NL4-3 was replaced with the Renilla Luciferase gene. The NL-RLuc virus was prepared by co-transfection of two plasmids, pNLRLuc and pVSVenv. The pNLRLuc contains the NL-Rluc DNA cloned into pUC18 at the PvuII site, while the pVSVenv contains the gene for VSV G protein linked to an LTR promoter. Transfections were performed at a 1:3 ratio of pNLRLuc to pVSVenv on 293T cells using the LipofectAMINE PLUS kit from Invitrogen (Carlsbad, Calif.) according to manufactures instruction, and the pseudotype virus generated was titered in MT-2 cells.
Susceptibility of viruses to compounds was determined by incubation in the presence of serial dilutions of the compound. The 50% effective concentration (EC50) was calculated by using the exponential form of the median effect equation where (Fa)=1/[1+(ED50/drug conc.)m] (Johnson V A, Byington R T. Infectivity Assay. In Techniques in HIV Research. ed. Aldovini A, Walker B D. 71-76. New York: Stockton Press. 1990). The anti-viral activity of compounds was evaluated under three serum conditions, 10% FBS, 15 mg/ml human serum albumin/10% FBS or 40% human serum/5% FBS, and the results from at least 2 experiments were used to calculate the EC50 values. Results are shown in the Table 2. Activity equal to A refers to a compound having EC50=0.003 to 0.10 μM while B and C denote compounds with EC50=0.1 to 1.0 μM and EC50≧1.0 μM respectively.










TABLE 2







Example
Activity



















1
B



2
A



3
C



4
A



5
A



6
A



7
A



8
A



9
A



10
A



11
A



12
A



13
A



14
A



15
A



16
A



17
A



18
A



19
A



20
A



21
A



22
B



23
A



24
C



25
C



26
A



27
A



28
B



29
A



30
A



31
A



32
A



33
A



34
A



35
A



36
A



37
B



38
A



39
A



40
A



41
A



42
A



43
A



44
A



45
A



46
A



47
A



48
B



49
A



50
A



51
A



52
A



53
B



54
A



55
A



56
A



57
A



58
A



59
B



60
A



61
B



62
A



63
A



64
C



65
A



66
B



67
A



68
C



69
A



70
B



71
B



72
B



73
A



74
A



75
A



76
B



77
A



78
A



79
B



80
B



81
A



82
A



83
C



84
A



85
A



86
A



87
A



88
A



89
A



90
A



91
A



92
A



93
A



94
A



95
A



96
A



97
A



98
A



99
A



100
A



101
A



102
A



103
B



104
B



105
A



106
A



107
A



108
A



109
A



110
A



111
A



113
A



114
A



115
A



116
A



117
A



118
A



119
A



120
A



121
A



122
A



123
A



124
A



125
A



126
A



127
A



128
A



129
A



130
A



132
A



133
B



134
A



135
A



136
A



137
B



138
A



139
A



140
A



141
C



142
B



144
C



146
A



147
B



148
B



149
B



150
B



151
B



152
B



153
B



154
B



155
B



156
B



157
A



158
A



159
A



160
B



161
B



162
A



163
B



164
B



165
A



166
C



167
B



168
A



169
A



170
A



171
B



172
B



173
A



174
A



175
B



176
A



177
A



178
A



179
B



180
C



181
A



187
B



188
B



189
A



190
A



191
B



192
B



193
C



194
B



195
B



196
A



197
C



198
B



199
B



200
A



201
B



202
A



203
A



204
A



205
A



206
A



207
A



208
A



209
B



210
A



211
A



212
A



213
A



214
B



215
A



216
A



217
A



218
A



219
A



220
A



221
A



222
A



223
A



224
B



225
A



226
A



227
B



228
B



229
A



230
A



231
A



232
A



233
A



234
A



235
A



236
B



237
A



238
A



239
A



240
A



241
A



242
A



243
A



244
A



245
A



246
A



247
A



248
A



249
A



250
A



251
A



252
C



253
C



254
A



255
A



256
B



257
A



258
A



259
A



260
A



262
A



263
B



265
B



266
A



267
A



268
A



270
A



273
B



279
A



281
A



282
A










Additional examples are shown in Table 2a.









TABLE 2a






HIV-Integrase
Inhibition of HIV


Example
Inhibition Activity
replication







284
A
A


285
NA
A


286
A
A


287
A
A


288
A
A


289
A
C


290
A
B


291
A
C


292
A
C


293
A
B


294
A
A


295
A
A


296
A
A


297
A
A


298
A
A


299
A
A


300
A
A


301
A
A


302
A
A


303
B
C


304
A
A


305
A
A


306
A
A


307
A
A


308
A
A


309
A
A


310
A
A


311
A
A


312
A
A


313
A
NA


314
A
A


315
A
A


316
A
A


317
A
A


318
A
A


319
A
A









Combination Studies
Example 19 demonstrated synergistic or additive-synergistic HIV antiviral activity when used in conjunction with a variety of other antiviral agents, as described below.
Virus and cell lines. The T-cell line, MT-2, was obtained through the AIDS Research and Reference Reagent Program. MT-2 cells were sub-cultured twice a week in RPMI 1640 medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, and 10 mM HEPES buffer pH 7.5. The HIV-1303B virus is a molecular clone derived from the NL4-3 strain of HIV-1 that was obtained from the NIH AIDS Research and Reference Reagent Program. For combinations with enfuvirtide, the NL36G virus was used. This NL4-3 derivative has the naturally occurring enfuvirtide resistance mutation in gp41 (36D) changed to a sensitive phenotype (D36G). Virus stocks were made by transfecting 293T cells with a proviral DNA clone using LipofectAMINE PLUS (Invitrogen), according to the manufacturer's instructions. Three days post-transfection, virus was harvested and passaged once in MT-2 cells before titration in MT-2 cells.
Chemicals. Example 19, atazanavir, didanosine, stavudine, efavirenz, and enfuvirtide (T-20) were synthesized by Bristol-Myers Squibb using published or known reactions. Amprenavir, indinavir, nelfmavir, nevirapine, lopinavir, lamivudine, ritonavir, tenofovir, saquinavir, delavirdine and abacavir were extracted from commercial formulations of the prescribed drugs and purified using published or common techniques. Tenofovir was tested as tenofovir disopoxil fumarate. Zidovudine and zalcitabine were purchased from Sigma, and emtricitabine from Moravek Biochemicals.
Drug Susceptibility and Cytotoxicity Assays. For drug susceptibility assays, MT-2 cells were infected with HIV-1 303B (or NL36G), at an MOI of 0.001, and seeded into 96-well microtiter plates (2.5×105 cells/ml) containing serial dilutions of test compounds. The drug combinations were set up using ratios of the two drugs of 1:1, 1:2.5 and 2.5:1 times the EC50 value determined for each drug in prior experiments. Each drug ratio consisted of an array of 3-fold serial dilutions, and was performed with eight or more replicas on separate multi-well plates. HIV infected cells were incubated at 37° C. in 5% CO2, and on day five post-infection, the extent of virus replication was measured by determining cell viability using the CellTiter 96 Aqueous Non-Radioactive Cell Proliferation Assay (Promega). Maximal cell protection was typically seen in samples treated with the highest drug concentration. In the cell viability assay, the tetrazolium compound MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-4-sulphophenyl-2H-tetrazolium) is added to cells, whereby enzymes in metabolically active cells convert it into a colored formazan product, which is quantitated by reading absorbance at 490 nm. Cytotoxicity assays were performed in parallel with the combination experiments. Here, uninfected cells were exposed to the same drug combinations, and assayed after five days for cell viability using the MTS assay.
Analysis of Drug Combination Effects. For determination of combination index (CI) values, drugs were diluted in a fixed ratio and multiple ratios were analyzed. The drug serial dilutions spanned a range of concentrations near the EC50 value of each compound, so that equivalent antiviral activities could be compared. The normalized responses from each therapy are fit to the four-parameter logistic model with a common minimum and maximum across all therapies. Conceptually, this equation can be written as




  
    
      Fa
      i
    
    =
    
      A
      +
      
        
          D
          -
          A
        
        
          1
          +
          
            
              (
              
                
                  
                    C
                    1
                    
                      I
                      1
                    
                  
                  ⁢
                  
                    C
                    2
                    
                      I
                      2
                    
                  
                  ⁢
                  
                    C
                    3
                    
                      I
                      3
                    
                  
                  ⁢
                  
                    C
                    4
                    
                      I
                      4
                    
                  
                  ⁢
                  
                    C
                    5
                    
                      I
                      5
                    
                  
                
                
                  concentration
                  i
                
              
              )
            
            
              
                B
                1
                
                  I
                  1
                
              
              ⁢
              
                B
                2
                
                  I
                  2
                
              
              ⁢
              
                B
                3
                
                  I
                  3
                
              
              ⁢
              
                B
                4
                
                  I
                  4
                
              
              ⁢
              
                B
                5
                
                  I
                  5
                
              
            
          
        
      
    
  
  ,
  
    

  
  ⁢
  
    
      where
      ⁢
      
        
      
      ⁢
      
        I
        j
      
    
    =
    
      {
      
        
          
            1
          
          
            
              
                if
                ⁢
                
                  
                
                ⁢
                therapy
              
              =
              j
            
          
        
        
          
            0
          
          
            otherwise
          
        
      
    
  




    
    
        for j=1, 2, 3, 4, or 5.
    
    


In this equation, Fa stands for “fraction affected,” and represents the fraction of the viral load that has been inactivated. For example, Fa of 0.75 indicates that viral replication had been inhibited by 75%, relative to the no-drug controls. The EC50, represented by the Ci in the above equation, is the drug concentration that is expected to reduce the amount of virus by 50%, and the Bi are the parameters that reflects the slope of the concentration-response curve. For this assay, A is the bottom plateau common to all curves, D is the common top plateau, Bj is the “slope” parameter for the jth therapy, and Cj is the concentration that produces an effect equal to the average of A and D for the jth therapy. Therapies 1 and 2 correspond to mono therapies 1 and 2, respectively. Therapies 3, 4, and 5 correspond to the three combination therapies. EC50 values for each drug were determined from the single drug experiments, using the above equation. The equation was fit using a nonlinear regression routine (Proc Nlin) in PC SAS version 8.2 (SAS Institute Inc.).
To assess antiviral effects of different drug combination treatments, combination indices (CIs) were calculated according to Chou and Rideout. The combination index was computed as




  CI
  =
  
    
      
        
          [
          D
          ]
        
        1
      
      
        
          [
          Dm
          ]
        
        1
      
    
    +
    
      
        
          [
          D
          ]
        
        2
      
      
        
          [
          Dm
          ]
        
        2
      
    
  




In this equation [Dm]1 and [Dm]2 are the concentrations of drugs that would individually produce a specific level of effect, while [D]1 and [D]2 are the concentrations of drugs in combination that would produce the same level of effect.
Theoretically, additivity is implied if the CI is equal to one, synergy if the CI is less than one, and antagonism if the CI is greater than one. However, extensive experience with combination studies indicates that there are inherent laboratory variables that must be taken into account in interpreting the CIs. At best, a range can be constructed that contains the likely values for the CI, given the noise in the data. In this report, these ranges are reported in parentheses next to each point estimate of the CI. For example, when a CI of “0.52 (0.36, 0.69)” is reported this means that the best estimate of the CI is 0.52, but due to noise in the data, values from 0.36 to 0.69 are also reasonable values for the CI. This range, 0.36 to 0.69 falls entirely below the value of 1.0, and hence all likely values for the Clare less than 1.0. Therefore, synergistic behavior can be inferred for this combination. If the range fell entirely above 1.0, we would infer antagonistic behavior. If the range were to include 1.0, we would infer additivity.
In carrying out the combination experiments, the EC50 for Example 19 and each comparator compound was determined during the course of each study, and used in the subsequent data analysis. The determined values are consistent with previously published data and are shown in Table 3.







TABLE 3







Anti-HIV Activity of Compounds in Two-Drug Combination Studies













Highest Drug



Compound
EC50 (μM)
Concentration (μM)















Example 19
0.022
2.5



Zidovudine
0.012
2.5



Didanosine
10.7
100



Stavudine
0.241
15



Lamivudine
0.203
15



Abacavir
1.04
30



Tenofovir
0.018
2.5



Emtricitabine
0.053
20



Zalcitabine
0.124
10



Efavirenz
0.0016
0.2



Nevirapine
0.095
5



Delavirdine
0.043
5



Ritonavir
0.048
2.5



Indinavir
0.026
2.5



Nelfinavir
0.022
2.5



Saquinavir
0.011
2.5



Amprenavir
0.062
2.5



Atazanavir
0.010
1



Lopinavir
0.017
2.5



Enfuvirtide
0.061
2.2










Two-Drug Combinations of Example 19 with Nucleoside Reverse Transcriptase Inhibitors. Eight nucleoside RT inhibitors (didanosine, stavudine, zidovudine, lamivudine, abacavir, zalcitabine, emtricitibine and the nucleoside phosphonate, tenofovir) were combined with Example 19 at a range of concentrations near the EC50 value of each compound, so that equivalent antiviral activities could be compared. All estimates were computed using SAS Proc NLIN, and a two-parameter logistic. Data is presented in Table 4 as the combination indices and the asymptotic confidence intervals for RT inhibitors at different molar ratios.
Four nucleoside RT inhibitors; didanosine, stavudine, abacavir and emtricitibine, show synergistic antiviral effects in combination with Example 19 at all effective levels and all molar ratios. The other four RT inhibitors: zidovudine, lamivudine, tenofovir and zalcitabine all have combination indices suggestive of synergistic behavior with Example 19. However, the upper ranges of the confidence intervals for some of the effective levels are greater than 1, so the overall effects of these compounds with Example 19 are classified as synergistic to additive. No significant antagonism of anti-HIV activity is observed. No enhanced cytotoxicity was encountered at the highest concentrations tested with any of the drug combinations, as measured by the XTT reduction assay.







TABLE 4







Two-Drug Combinations using Example 19 and Nucleoside Reverse


Transcriptase Inhibitors.









Combination Indices at % HIV Inhibitionb


Molar Ratio
(Confidence Interval)










(EC50 Ratio)a
50%
75%
90%





Zidovudine





1:1 (1:1)
0.59 (0.49, 0.69)
0.56 (0.41, 0.70)
0.54 (0.36, 0.70)


2:5 (1:2.5)
0.65 (0.56, 0.75)
0.73 (0.59, 0.86)
0.81 (0.57, 1.06)


5:2 (2.5:1)
0.72 (0.61, 0.83)
0.83 (0.64, 1.02)
0.96 (0.63, 1.30)


Didanosine


1:1000 (1:1)
0.58 (0.53, 0.63)
0.47 (0.35, 0.60)
0.39 (0.22, 0.56)


1:2500 (1:2.5)
0.56 (0.51, 0.62)
0.46 (0.40, 0.52)
0.38 (0.32, 0.44)


1:400 (2.5:1)
0.50 (0.46, 0.54)
0.49 (0.44, 0.53)
0.48 (0.42, 0.54)


Stavudine


1:6 (1:1)
0.52 (0.43, 0.61)
0.46 (0.36, 0.56)
0.42 (0.26, 0.57)


1:15 (1:2.5)
0.60 (0.48, 0.71)
0.60 (0.46, 0.74)
0.61 (0.40, 0.83)


5:12 (2.5:1)
0.70 (0.57, 0.84)
0.63 (0.48, 0.79)
0.58 (0.37, 0.81)


Lamivudine


1:6 (1:1)
0.80 (0.63, 0.98)
0.80 (0.55, 1.05)
0.81 (0.42, 1.21)


1:15 (1:2.5)
1.09 (0.88, 1.31)
1.00 (0.72, 1.27)
0.92 (0.52, 1.33)


5:12 (2.5:1)
0.94 (0.80, 1.08)
0.84 (0.65, 1.03)
0.75 (0.49, 1.02)


Abacavir


1:30 (1:1)
0.68 (0.59, 0.76)
0.67 (0.56, 0.79)
0.67 (0.49, 0.84)


1:75 (1:2.5)
0.87 (0.77, 0.97)
0.74 (0.63, 0.85)
0.63 (0.50, 0.77)


1:12 (2.5:1)
0.86 (0.76, 0.96)
0.82 (0.68, 0.96)
0.79 (0.58, 0.99)


Tenofovir


1:1 (1:1)
0.66 (0.57, 0.75)
0.54 (0.45, 0.62)
0.43 (0.33, 0.53)


2:5 (1:2.5)
0.68 (0.59, 0.77)
0.60 (0.48, 0.72)
0.52 (0.36, 0.69)


5:2 (2.5:1)
0.90 (0.77, 1.04)
0.84 (0.67, 1.01)
0.79 (0.55, 1.03)


Zalcitabine


1:4 (1:1)
0.61 (0.48, 0.73)
0.56 (0.39, 0.73)
0.53 (0.28, 0.79)


1:10 (1:2.5)
0.55 (0.44, 0.66)
0.40 (0.31, 0.49)
0.30 (0.18, 0.42)


5:8 (2.5:1)
0.80 (0.62, 0.97)
0.82 (0.58, 1.07)
0.86 (0.46, 1.26)


Emtricitabine


1:8 (1:1)
0.31 (0.27, 0.36)
0.23 (0.18, 0.28)
0.18 (0.12, 0.24)


1:20 (1:2.5)
0.31 (0.26, 0.37)
0.23 (0.19, 0.29)
0.17 (0.12, 0.28)


5:16 (2.5:1)
0.33 (0.28, 0.38)
0.25 (0.20, 0.30)
0.19 (0.13, 0.25)





aRatio of Example 19 to comparator compound


bA lower bound of the asymptotic confidence interval greater than 1 indicates antagonisms, an upper bound of less than 1 indicates synergism and a value of 1 being contained in the interval indicates additivity. The 95% confidence intervals are shown in parenthesis, and represent a measure of variability in the data.






Two-Drug Combinations of Example 19 with Non-Nucleoside Reverse Transcriptase Inhibitors. Three non-nucleoside RT inhibitors were combined with Example 19 at a range of concentrations near the EC50 value of each compound, as described above for nucleoside RT inhibitors. Data is presented in Table 5 as the combination indices and the asymptotic confidence intervals at different molar ratios. Of the three, nevaripine shows strong synergistic effects in combination with Example 19. Synergy is seen at all effective concentrations and at all molar ratios. Sustiva and nevirapine also exhibit combination indices indicative of synergism at most effective concentrations and molar ratios. However, in some cases, the upper range of the asymptotic confidence interval is greater than 1, so an additive effect can not be ruled out. No enhanced cytotoxicity was observed at the highest concentrations tested with any of the drug combinations, suggesting a potential for therapeutic efficacy of Example 19 combinations with non-nucleoside RT inhibitors.







TABLE 5







Two-Drug Combinations using Example 19 and Non-Nucleoside


Reverse Transcriptase Inhibitors.









Combination Indices at % HIV Inhibitionb


Molar Ratio
(Confidence Interval)










(EC50 Ratio)a
50%
75%
90%





Efavirenz





5:2 (1:1)
0.83 (0.75, 0.92)
0.76 (0.64, 0.88)
0.70 (0.54, 0.85)


1:1 (1:2.5)
0.95 (0.86, 1.04)
0.93 (0.85, 1.02)
0.93 (0.79, 1.06)


25:4 (2.5:1)
1.01 (0.92, 1.10)
0.96 (0.83, 1.10)
0.94 (0.73, 1.14)


Nevirapine


1:2 (1:1)
0.72 (0.62, 0.82)
0.69 (0.55, 0.83)
0.66 (0.46, 0.87)


1:5 (1:2.5)
0.76 (0.64, 0.87)
0.81 (0.64, 0.97)
0.87 (0.59, 1.15)


5:4 (2.5:1)
0.73 (0.62, 0.83)
0.68 (0.55, 0.81)
0.64 (0.45, 0.84)


Delavirdine


1:2 (1:1)
0.76 (0.64, 0.89)
0.60 (0.47, 0.74)
0.48 (0.31, 0.64)


1:5 (1:2.5)
0.68 (0.58, 0.77)
0.44 (0.32, 0.54)
0.28 (0.17, 0.39)


5:4 (2.5:1)
0.80 (0.68, 0.92)
0.53 (0.42, 0.63)
0.35 (0.25, 0.45)





aRatio of Example 19 to comparator compound.


bA lower bound of the asymptotic confidence interval greater than 1 indicates antagonisms, an upper bound of less than 1 indicates synergism, and a value of 1 being contained in the interval indicates additivity. The 95% confidence intervals are shown in parenthesis, and represent a measure of variability in the data.






Two-Drug Combinations Involving Example 19 and HIV Protease Inhibitors. Evaluation of Example 19 for drug combination therapy with protease inhibitors was carried out using indinavir, amprenavir, nelfinavir, lopinavir, saquinavir, ritonavir and atazanavir. Results from this two-drug combination study are summarized in Table 6. Again, the combination indices observed with Example 19 and all protease inhibitors at almost all effective levels and molar ratios are suggestive of a synergistic relationship. This is especially true for saquinavir and atazanavir, where the confidence interval is below one at all concentrations and effective levels. Meanwhile, the upper range of the confidence interval is greater than one in only one condition for ritonavir, indinavir and lopinavir, so an additive relationship with Example 19 cannot be ruled out. In addition, the upper range of the confidence interval for nelfinavir and amprenavir are slightly greater than 1 under a few conditions, suggestive of a synergistic-additive effect for these compounds with Example 19. No cytotoxicity was observed at the highest concentrations used in any of these combination antiviral assays.







TABLE 6







Two-Drug Combination using Example 19 and Protease Inhibitors.









Combination Indices at % HIV Inhibitionb


Molar Ratio
(Confidence Interval)










(EC50 Ratio)a
50%
75%
90%





Indinavir





1:1 (1:1)
0.86 (0.80, 0.92)
0.71 (0.62, 0.81)
0.60 (0.46, 0.73)


2:5 (1:2.5)
0.92 (0.84, 0.99)
0.84 (0.76, 0.92)
0.77 (0.66, 0.89)


5:2 (2.5:1)
0.94 (0.87, 1.02)
0.79 (0.71, 0.87)
0.67 (0.57, 0.77)


Nelfinavir


1:1 (1:1)
0.79 (0.73, 0.86)
0.75 (0.67, 0.82)
0.71 (0.56, 0.84)


2:5 (1:2.5)
0.97 (0.89, 1.06)
0.87 (0.79, 0.95)
0.78 (0.68, 0.88)


5:2 (2.5:1)
1.09 (1.00, 1.18)
0.98 (0.89, 1.08)
0.90 (0.76, 1.03)


Saguinavir


1:1 (1:1)
0.77 (0.70, 0.84)
0.67 (0.58, 0.75)
0.58 (0.47, 0.69)


2:5 (1:2.5)
0.43 (0.38, 0.48)
0.51 (0.42, 0.59)
0.59 (0.44, 0.74)


5:2 (2.5:1)
0.81 (0.72, 0.89)
0.77 (0.66, 0.89)
0.74 (0.57, 0.91)


Amprenavir


1:1 (1:1)
0.83 (0.67, 1.00)
0.84 (0.61, 1.08)
0.89 (0.50, 1.23)


2:5 (1:2.5)
0.84 (0.69, 0.99)
0.77 (0.58, 0.96)
0.75 (0.46, 1.04)


5:2 (2.5:1)
0.90 (0.77, 1.04)
0.62 (0.50, 0.74)
0.44 (0.30, 0.57)


Atazanavir


5:2 (1:1)
0.87 (0.79, 0.96)
0.67 (0.55, 0.80)
0.52 (0.37, 0.67)


1:1 (1:2.5)
0.65 (0.51, 0.78)
0.45 (0.31, 0.58)
0.31 (0.19, 0.44)


25:4 (2.5:1)
0.78 (0.69, 0.86)
0.60 (0.54, 0.67)
0.47 (0.40, 0.54)


Lopinavir


1:1 (1:1)
0.74 (0.67, 0.82)
0.77 (0.66, 0.88)
0.84 (0.65, 1.02)


2:5 (1:2.5)
0.77 (0.66, 0.88)
0.56 (0.36, 0.75)
0.41 (0.19, 0.64)


5:2 (2.5:1)
0.76 (0.69, 0.83)
0.62 (0.55, 0.70)
0.54 (0.47, 0.61)


Ritonavir


1:1 (1:1)
0.80 (0.67, 0.93)
0.57 (0.41, 0.73)
0.44 (0.23, 0.65)


2:5 (1:2.5)
0.73 (0.59, 0.88)
0.64 (0.48, 0.80)
0.61 (0.38, 0.85)


5:2 (2.5:1)
0.92 (0.72, 1.11)
0.72 (0.50, 0.93)
0.59 (0.32, 0.87)





aRatio of Example 19 to comparator compound.


bA lower bound of the asymptotic confidence interval greater than 1 indicates antagonisms, an upper bound of less than 1 indicates synergism, and a value of 1 being contained in the interval indicates additivity. The 95% confidence intervals are shown in parenthesis, and represent a measure of variability in the data.






Two-Drug Combination of Example 19 with Enfuvirtide. Enfuvirtide (T-20) is an HIV gp41 fusion inhibitor and the first approved entry class inhibitor. The results presented in Table 7 indicate that the combination of Example 19 with T-20 is synergistic. No significant cytotoxicity was observed at the highest concentration of the combined drugs.







TABLE 7







Two-Drug Combination study of Example 19 with Enfuvirtide.









Combination Indices at % HIV Inhibitionb


Molar Ratio
(Confidence Interval)










(EC50 Ratio)a
50%
75%
90%





Enfuvirtide





5:44 (1:1)
0.68 (0.59, 0.77)
0.59 (0.48, 0.70)
0.52 (0.37, 0.67)


1:22 (1:2.5)
0.75 (0.65, 0.85)
0.60 (0.49, 0.70)
0.48 (0.34, 0.61)


25:88 (2.5:1)
0.76 (0.65, 0.86)
0.73 (0.59, 0.86)
0.71 (0.50, 0.92)





aRatio of Example 19 to comparator compound.


bA lower bound of the asymptotic confidence interval greater than 1 indicates antagonisms, an upper bound of less than 1 indicates synergism, and a value of 1 being contained in the interval indicates additivity. The 95% confidence intervals are shown in parenthesis, and represent a measure of variability in the data.






Pharmaceutical Composition and Methods of Use
The compounds of this invention inhibit HIV integrase. HIV integrase inhibitors belonging to a class of diketo acid compounds prevented viral integration and inhibited HIV-1 replication in cells (Hazuda et al. Science 2000, 287, 646). Recently, HIV integrase inhibitors have been accepted into clinical trials for treating AIDS and HIV infection (Neamati Expert. Opin. Ther. Patents 2002, 12, 709, Pais and Burke Drugs Fut. 2002, 27, 1101).
Accordingly, another aspect of the invention is a method for treating HIV infection in a human patient comprising administering a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, with a pharmaceutically acceptable carrier.
Another aspect of the invention is a method for treating HIV infection in a human patient comprising the administration of a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, with a therapeutically effective amount of at least one other agent used for treatment of AIDS or HIV infection selected from the group consisting of nucleoside HIV reverse transcriptase inhibitors, non-nucleoside HIV reverse transcriptase inhibitors, HIV protease inhibitors, HIV fusion inhibitors, HIV attachment inhibitors, CCR5 inhibitors, CXCR4 inhibitors, HIV budding or maturation inhibitors, and HIV integrase inhibitors.
Another aspect of the invention is a method wherein the agent is a nucleoside HIV reverse transcriptase inhibitor.
Another aspect of the invention is a method wherein the nucleoside HIV reverse transcriptase inhibitor is selected from the group consisting of abacavir, didanosine, emtricitabine, lamivudine, stavudine, tenofovir, zalcitabine, and zidovudine, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is a method wherein the agent is a non-nucleoside HIV reverse transcriptase inhibitor.
Another aspect of the invention is a method wherein the non-nucleoside HIV reverse transcriptase inhibitor is selected from the group consisting of delavirdine, efavirenz, and nevirapine, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is a method wherein the agent is an HIV protease inhibitor.
Another aspect of the invention is a method wherein the HIV protease inhibitor is selected from the group consisting of amprenavir, atazanavir, indinavir, lopinavir, nelfmavir, ritonavir, saquinavir and fosamprenavir, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is a method wherein the agent is an HIV fusion inhibitor.
Another aspect of the invention is a method wherein the HIV fusion inhibitor is enfuvirtide or T-1249, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is a method wherein the agent is an HIV attachment inhibitor.
Another aspect of the invention is a method wherein the agent is a CCR5 inhibitor.
Another aspect of the invention is a method wherein the CCR5 inhibitor is selected from the group consisting of Sch-C, Sch-D, TAK-220, PRO-140, and UK-427,857, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is a method wherein the agent is a CXCR4 inhibitor.
Another aspect of the invention is a method wherein the CXCR4 inhibitor is AMD-3100, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is a method wherein the agent is an HIV budding or maturation inhibitor.
Another aspect of the invention is a method wherein the budding or maturation inhibitor is PA-457, or a pharmaceutically acceptable salt, or solvate thereof.
Another aspect of the invention is a method wherein the agent is an HIV integrase inhibitor.
Another aspect of the invention is a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, with at least one other agent used for treatment of AIDS or HIV infection selected from the group consisting of nucleoside HIV reverse transcriptase inhibitors, non-nucleoside HIV reverse transcriptase inhibitors, HIV protease inhibitors, HIV fusion inhibitors, HIV attachment inhibitors, CCR5 inhibitors, CXCR4 inhibitors, HIV budding or maturation inhibitors, and HIV integrase inhibitors, and a pharmaceutically acceptable carrier.
Another aspect of the invention is the composition wherein the agent is a nucleoside HIV reverse transcriptase inhibitor.
Another aspect of the invention is the composition wherein the nucleoside HIV transcriptase inhibitor is selected from the group consisting of abacavir, didanosine, emtricitabine, lamivudine, stavudine, tenofovir, zalcitabine, and zidovudine, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is the composition wherein the agent is a non-nucleoside HIV reverse transcriptase inhibitor.
Another aspect of the invention is the composition wherein the non-nucleoside HIV reverse transcriptase inhibitor is selected from the group consisting of delavirdine, efavirenz, and nevirapine, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is the composition wherein the agent is an HIV protease inhibitor.
Another aspect of the invention is the composition wherein the HIV protease inhibitor is selected from the group consisting of amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, saquinavir and fosamprenavir, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is the composition wherein the agent is an HIV fusion inhibitor.
Another aspect of the invention is the composition method wherein the HIV fusion inhibitor is enfuvirtide or T-1249, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is the composition wherein the agent is an HIV attachment inhibitor.
Another aspect of the invention is the composition wherein the agent is a CCR5 inhibitor.
Another aspect of the invention is the composition wherein the CCR5 inhibitor is selected from the group consisting of Sch-C, Sch-D, TAK-220, PRO-140, and UK-427,857, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is a method wherein the agent is a CXCR4 inhibitor.
Another aspect of the invention is a method wherein the CXCR4 inhibitor is AMD-3100 or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is the composition wherein the agent is an HIV budding or maturation inhibitor.
Another aspect of the invention is the composition wherein the budding or maturation inhibitor is PA-457, or a pharmaceutically acceptable salt or solvate thereof.
Another aspect of the invention is the composition wherein the agent is an HIV integrase inhibitor.
“Combination,” “coadministration,” “concurrent,” and similar terms referring to the administration of a compound of Formula I with at least one anti-HIV agent mean that the components are part of a combination antiretroviral therapy or highly active antiretroviral therapy (HAART) as understood by practitioners in the field of AIDS and HIV infection.
“Therapeutically effective” means the amount of agent required to provide a meaningful patient benefit as understood by practitioners in the field of AIDS and HIV infection. In general, the goals of treatment are suppression of viral load, restoration and preservation of immunologic function, improved quality of life, and reduction of HIV-related morbidity and mortality.
“Patient” means a person infected with the HIV virus and suitable for therapy as understood by practitioners in the field of AIDS and HIV infection.
“Treatment,” “therapy,” “regimen,” “HIV infection,” “ARC,” “AIDS” and related terms are used as understood by practitioners in the field of AIDS and HIV infection.
The compounds of this invention are generally given as pharmaceutical compositions comprised of a therapeutically effective amount of a compound of Formula I or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier and may contain conventional excipients. A therapeutically effective amount is that which is needed to provide a meaningful patient benefit. Pharmaceutically acceptable carriers are those conventionally known carriers having acceptable safety profiles. Compositions encompass all common solid and liquid forms including capsules, tablets, losenges, and powders as well as liquid suspensions, syrups, elixers, and solutions. Compositions are made using common formulation techniques, and conventional excipients (such as binding and wetting agents) and vehicles (such as water and alcohols) are generally used for compositions.
Solid compositions are normally formulated in dosage units and compositions providing from about 1 to 1000 mg of the active ingredient per dose are preferred. Some examples of dosages are 1 mg, 10 mg, 100 mg, 250 mg, 500 mg, and 1000 mg. Generally, other antiretroviral agents will be present in a unit range similar to agents of that class used clinically. Typically, this is 0.25-1000 mg/unit.
Liquid compositions are usually in dosage unit ranges. Generally, the liquid composition will be in a unit dosage range of 1-100 mg/mL. Some examples of dosages are 1 mg/mL, 10 mg/mL, 25 mg/mL, 50 mg/mL, and 100 mg/mL. Generally, other antiretroviral agents will be present in a unit range similar to agents of that class used clinically. Typically, this is 1-100 mg/mL.
The invention encompasses all conventional modes of administration; oral and parenteral methods are preferred. Generally, the dosing regimen will be similar to other antiretroviral agents used clinically. Typically, the daily dose will be 1-100 mg/kg body weight daily. Generally, more compound is required orally and less parenterally. The specific dosing regime, however, will be determined by a physician using sound medical judgement.
The invention also encompasses methods where the compound is given in combination therapy. That is, the compound can be used in conjunction with, but separately from, other agents useful in treating AIDS and HIV infection. Some of these agents include HIV attachment inhibitors, CCR5 inhibitors, CXCR4 inhibitors, HIV cell fusion inhibitors, HIV integrase inhibitors, HIV nucleoside reverse transcriptase inhibitors, HIV non-nucleoside reverse transcriptase inhibitors, HIV protease inhibitors, budding and maturation inhibitors, immunomodulators, and anti-infectives. In these combination methods, the compound of Formula I will generally be given in a daily dose of 1-100 mg/kg body weight daily in conjunction with other agents. The other agents generally will be given in the amounts used therapeutically. The specific dosing regime, however, will be determined by a physician using sound medical judgement.
Table 8 lists some agents useful in treating AIDS and HIV infection which are suitable for this invention.









TABLE 8





DRUG NAME
MANUFACTURER
INDICATION















ANTIVIRALS









097
Hoechst/Bayer
HIV infection, AIDS,


(non-nucleoside reverse

ARC


transcriptase inhibitor)


Amprenavir
Glaxo Wellcome
HIV infection, AIDS,


141 W94

ARC


GW 141


(protease inhibitor)


Abacavir (1592U89)
Glaxo Wellcome
HIV infection, AIDS,


GW 1592

ARC


(RT inhibitor)


Acemannan
Carrington Labs
ARC



(Irving, TX)


Acyclovir
Burroughs Wellcome
HIV infection, AIDS,




ARC, in combination




with AZT


AD-439
Tanox Biosystems
HIV infection, AIDS,




ARC


AD-519
Tanox Biosystems
HIV infection, AIDS,




ARC


Adefovir dipivoxil
Gilead Sciences
HIV infection, ARC,


AL-721
Ethigen
PGL HIV positive,



(Los Angeles, CA)
AIDS


Alpha Interferon
Glaxo Wellcome
Kaposi's sarcoma


HIV in combination


w/Retrovir


Ansamycin
Adria Laboratories
ARC


LM 427
(Dublin, OH)



Erbamont



(Stamford, CT)


Antibody which
Advanced Biotherapy
AIDS, ARC


Neutralizes pH
Concepts


Labile alpha aberrant
(Rockville, MD)


Interferon


AR177
Aronex Pharm
HIV infection, AIDS,




ARC


Beta-fluoro-ddA
Nat'l Cancer Institute
AIDS-associated




diseases


BMS-232623
Bristol-Myers Squibb/
HIV infection, AIDS,


(CGP-73547)
Novartis
ARC


(protease inhibitor)


BMS-234475
Bristol-Myers Squibb/
HIV infection, AIDS,


(CGP-61755)
Novartis
ARC


(protease inhibitor)


CI-1012
Warner-Lambert
HIV-1 infection


Cidofovir
Gilead Science
CMV retinitis, herpes,




papillomavirus


Curdlan sulfate
AJI Pharma USA
HIV infection


Cytomegalovirus
MedImmune
CMV retinitis


Immune globin


Cytovene
Syntex
Sight threatening


Ganciclovir

CMV peripheral,




CMV retinitis


Delaviridine
Pharmacia-Upjohn
HIV infection, AIDS,


(RT inhibitor)

ARC


Dextran Sulfate
Ueno Fine Chem.
AIDS, ARC, HIV



Ind. Ltd. (Osaka,
positive asymptomatic



Japan)


ddC
Hoffman-La Roche
HIV infection, AIDS,


Dideoxycytidine

ARC


ddI
Bristol-Myers Squibb
HIV infection, AIDS,


Dideoxyinosine

ARC; combination




with AZT/d4T


DMP-450
AVID
HIV infection, AIDS,


(protease inhibitor)
(Camden, NJ)
ARC


Efavirenz
DuPont Merck
HIV infection, AIDS,


(DMP 266)

ARC


(−)6-Chloro-4-(S)-


cyclopropylethynyl-


4(S)-trifluoro-


methyl-1,4-dihydro-


2H-3,1-benzoxazin-


2-one, STOCRINE


(non-nucleoside RT


inhibitor)


EL10
Elan Corp, PLC
HIV infection



(Gainesville, GA)


Famciclovir
Smith Kline
herpes zoster, herpes




simplex


FTC
Emory University
HIV infection, AIDS,


(reverse transcriptase

ARC


inhibitor)


GS 840
Gilead
HIV infection, AIDS,


(reverse transcriptase

ARC


inhibitor)


HBY097
Hoechst Marion
HIV infection, AIDS,


(non-nucleoside reverse
Roussel
ARC


transcriptase inhibitor)


Hypericin
VIMRx Pharm.
HIV infection, AIDS,




ARC


Recombinant Human
Triton Biosciences
AIDS, Kaposi's


Interferon Beta
(Almeda, CA)
sarcoma, ARC


Interferon alfa-n3
Interferon Sciences
ARC, AIDS


Indinavir
Merck
HIV infection, AIDS,




ARC, asymptomatic




HIV positive, also in




combination with




AZT/ddI/ddC


ISIS 2922
ISIS Pharmaceuticals
CMV retinitis


KNI-272
Nat'l Cancer Institute
HIV-associated




diseases


Lamivudine, 3TC
Glaxo Wellcome
HIV infection, AIDS,


(reverse transcriptase

ARC, also with AZT


inhibitor)


Lobucavir
Bristol-Myers Squibb
CMV infection


Nelfinavir
Agouron
HIV infection, AIDS,


(protease inhibitor)
Pharmaceuticals
ARC


Nevirapine
Boeheringer
HIV infection, AIDS,


(RT inhibitor)
Ingleheim
ARC


Novapren
Novaferon Labs, Inc.
HIV inhibitor



(Akron, OH)


Peptide T
Peninsula Labs
AIDS


Octapeptide
(Belmont, CA)


Sequence


Trisodium
Astra Pharm.
CMV retinitis, HIV


Phosphonoformate
Products, Inc.
infection, other CMV




infections


PNU-140690
Pharmacia Upjohn
HIV infection, AIDS,


(protease inhibitor)

ARC


Probucol
Vyrex
HIV infection, AIDS


RBC-CD4
Sheffield Med.
HIV infection, AIDS,



Tech (Houston, TX)
ARC


Ritonavir
Abbott
HIV infection, AIDS,


(protease inhibitor)

ARC


Saquinavir
Hoffmann-
HIV infection, AIDS,


(protease inhibitor)
LaRoche
ARC


Stavudine; d4T
Bristol-Myers Squibb
HIV infection, AIDS,


Didehydrodeoxy-

ARC


thymidine


Valaciclovir
Glaxo Wellcome
Genital HSV &




CMV infections


Virazole
Viratek/ICN
asymptomatic HIV-


Ribavirin
(Costa Mesa, CA)
positive, LAS, ARC


VX-478
Vertex
HIV infection, AIDS,




ARC


Zalcitabine
Hoffmann-LaRoche
HIV infection, AIDS,




ARC, with AZT


Zidovudine; AZT
Glaxo Wellcome
HIV infection, AIDS,




ARC, Kaposi's




sarcoma, in




combination




with other therapies


Tenofovir disoproxil,
Gilead
HIV infection, AIDS


fumarate salt (Viread ®)


(reverse transcriptase


inhibitor)


Combivir ®
GSK
HIV infection, AIDS


(reverse transcriptase


inhibitor)


abacavir succinate
GSK
HIV infection, AIDS


(or Ziagen ®)


(reverse transcriptase


inhibitor)


Reyataz ®
Bristol-Myers Squibb
HIV infection, AIDS


(atazanavir)


Fuzeon
Roche/Trimeris
HIV infection, AIDS,


(Enfuvirtide, T-20)

viral fusion inhibitor


Trizivir ®

HIV infection, AIDS


Kaletra ®
Abbott
HIV infection, AIDS,




ARC







IMMUNOMODULATORS









AS-101
Wyeth-Ayerst
AIDS


Bropirimine
Pharmacia Upjohn
Advanced AIDS


Acemannan
Carrington Labs, Inc.
AIDS, ARC



(Irving, TX)


CL246,738
American Cyanamid
AIDS, Kaposi's



Lederle Labs
sarcoma


EL10
Elan Corp, PLC
HIV infection



(Gainesville, GA)


FP-21399
Fuki ImmunoPharm
Blocks HIV fusion




with CD4+ cells


Gamma Interferon
Genentech
ARC, in combination




w/TNF (tumor




necrosis factor)


Granulocyte
Genetics Institute
AIDS


Macrophage Colony
Sandoz


Stimulating Factor


Granulocyte
Hoechst-Roussel
AIDS


Macrophage Colony
Immunex


Stimulating Factor


Granulocyte
Schering-Plough
AIDS, combination


Macrophage Colony

w/AZT


Stimulating Factor


HIV Core Particle
Rorer
Seropositive HIV


Immunostimulant


IL-2
Cetus
AIDS, in combination


Interleukin-2

w/AZT


IL-2
Hoffman-LaRoche
AIDS, ARC, HIV, in


Interleukin-2
Immunex
combination w/AZT


IL-2
Chiron
AIDS, increase in CD4


Interleukin-2

cell counts


(aldeslukin)


Immune Globulin
Cutter Biological
Pediatric AIDS, in


Intravenous
(Berkeley, CA)
combination w/AZT


(human)


IMREG-1
Imreg
AIDS, Kaposi's



(New Orleans, LA)
sarcoma, ARC, PGL


IMREG-2
Imreg
AIDS, Kaposi's



(New Orleans, LA)
sarcoma, ARC, PGL


Imuthiol Diethyl
Merieux Institute
AIDS, ARC


Dithio Carbamate


Alpha-2
Schering Plough
Kaposi's sarcoma


Interferon

w/AZT, AIDS


Methionine-
TNI Pharmaceutical
AIDS, ARC


Enkephalin
(Chicago, IL)


MTP-PE
Ciba-Geigy Corp.
Kaposi's sarcoma


Muramyl-Tripeptide
Amgen
AIDS, in combination


Granulocyte

w/AZT


Colony Stimulating


Factor


Remune
Immune Response
Immunotherapeutic



Corp.


rCD4
Genentech
AIDS, ARC


Recombinant


Soluble Human CD4


rCD4-IgG

AIDS, ARC


hybrids


Recombinant
Biogen
AIDS, ARC


Soluble Human CD4


Interferon
Hoffman-La Roche
Kaposi's sarcoma,


Alfa 2a
in combination w/AZT
AIDS, ARC


SK&F106528
Smith Kline
HIV infection


Soluble T4


Thymopentin
Immunobiology
HIV infection



Research Institute



(Annandale, NJ)


Tumor Necrosis
Genentech
ARC, in combination


Factor; TNF

w/gamma Interferon







ANTI-INFECTIVES









Clindamycin with
Pharmacia Upjohn
PCP


Primaquine


Fluconazole
Pfizer
Cryptococcal




meningitis,




candidiasis


Pastille
Squibb Corp.
Prevention of oral


Nystatin Pastille

candidiasis


Ornidyl
Merrell Dow
PCP


Eflornithine


Pentamidine
LyphoMed
PCP treatment


Isethionate (IM & IV)
(Rosemont, IL)


Trimethoprim

Antibacterial


Trimethoprim/sulfa

Antibacterial


Piritrexim
Burroughs Wellcome
PCP treatment


Pentamidine
Fisons Corporation
PCP prophylaxis


Isethionate for


Inhalation


Spiramycin
Rhone-Poulenc
Cryptosporidial



diarrhea


Intraconazole-
Janssen-Pharm.
Histoplasmosis;


R51211

cryptococcal




meningitis


Trimetrexate
Warner-Lambert
PCP


Daunorubicin
NeXstar, Sequus
Kaposi's sarcoma


Recombinant Human
Ortho Pharm. Corp.
Severe anemia assoc.


Erythropoietin

with AZT therapy


Recombinant Human
Serono
AIDS-related wasting,


Growth Hormone

cachexia


Megestrol Acetate
Bristol-Myers Squibb
Treatment of anorexia




assoc. W/AIDS


Testosterone
Alza, Smith Kline
AIDS-related wasting


Total Enteral
Norwich Eaton
Diarrhea and


Nutrition
Pharmaceuticals
malabsorption related




to AIDS









DESCRIPTION OF SPECIFIC EMBODIMENTS
Intermediate 1




2-(2-(Methylthio)ethoxy)acetic acid. A solution of 2-methylthioethanol (10.0 g, 0.108 mol) in dry tetrahydrofuran (25 ml) was added dropwise, over 30 min, to a suspension of sodium hydride (9.54 g of a 60% dispersion in mineral oil, 0.238 mol, washed twice with hexane) in dry tetrahydrofuran (250 ml) at 22° C. After 30 min, a solution of chloroacetic acid (10.25 g, 0.108 mol) in dry tetrahydrofuran (20 ml) was added dropwise, over 30 min at 22° C., and the resulting mixture was then heated under reflux for 5 h. The cooled mixture was treated with 250 ml of 1 N hydrochloric acid and sodium chloride added to the aqueous phase until saturation. The organic phase was separated and the aqueous phase washed with ethyl acetate. The combined organic extracts were washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Distillation of the resulting residue in vacuo gave 11.27 g (69% yield) of the title acid as a clear oil; bp 85-95° C./0.3 torr (bulb to bulb distillation, air bath temperature). 1HNMR 400 MHz (CDCl3) δ ppm: 2.18 (3H, s, SCH3), 2.76 (2H, t, J=6.6 Hz, CH2), 3.77 (2H, t, J=6.6 Hz, CH2), 4.20 (2H, s, OCH2).
Intermediate 2




Methyl 2-(2-(methylthio)ethoxy)acetate. A solution of intermediate 1,2-(2-(methylthio)ethoxy)acetic, (11.27 g, 0.075 mol) in dry dichloromethane (50 ml) was treated with oxalylchloride (13.0 ml, 0.15 mol) followed by a drop of N,N-dimethylformamide and the resulting mixture stirred at 22° C. for 5 h. The solvent and excess reagent were then evaporated under reduced pressure and the residual acid chloride was added dropwise to a cold mixture (0-5° C.) of methanol (30 ml) and pyridine (10 ml) in dichloromethane (50 ml). After 1 h at 22° C., the solvent was evaporated under reduced pressure. The resulting residue was diluted with ethyl acetate, washed with 1 N hydrochloric acid, saturated sodium bicarbonate and brine, dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. Distillation of the resulting residue in vacuo gave 11.42 g (93% yield) of the title ester as a clear oil; bp 65-75° C./0.1 torr (bulb to bulb distillation, air bath temperature). 1HNMR 400 MHz (CDCl3) δ ppm: 2.17 (3H, s, SCH3), 2.75 (2H, t, J=6.9 Hz, CH2), 3.74 (2H, t, J=6.9 Hz, CH2), 3.77 (3H, s, OCH3), 4.15 (2H, s, OCH2).
Intermediate 3




2-(2-(Methylthio)ethoxy)acetamidine hydrochloride salt. Intermediate 2, methyl 2-(2-(methylthio)ethoxy)acetate, (4.69 g, 28.6 mmol) was added to a solution of methylchloroaluminum amide (H. Geilen, C. Alonso-Alija, M. Hendrix, U. Niewohner and D. Schauss, Tetrahedron Lett., 2002, 43, 419-421)(0.114 mol; prepared in toluene (50 ml) from ammonium chloride 6.30 g (0.117 mol) and 57.0 ml (0.114 mol) of a 2 M solution of trimethylaluminum in toluene) and the resulting mixture was heated at 80° C. for 18 h. The reaction mixture was then cooled to 0° C., treated dropwise with methanol (100 ml) and stirred for another hour at 25° C. The solid which was formed was filtered and washed with methanol (300 ml). The combined filtrate was concentrated to give a white paste which was diluted with isopropanol (160 ml) and acetone (40 ml) and stirred at 25° C. for 1 h. The solid was then filtered off and the filtrate concentrated in vacuo to give 3.50 g (62% yield) of the title compound as an oil. 1HNMR 400 MHz (DMSO-d6) δ ppm: 2.10 (3H, s, SCH3), 2.71 (2H, t, J=6.8 Hz, CH2), 3.66 (2H, t, J=6.8 Hz, CH2), 4.34 (2H, s, OCH2). MS (ESI+) m/z 149 [M+H+].
Intermediate 4




Ethyl 5-benzyloxy-2-{(2-(methylthio)ethoxy)methyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate. Diethyl oxalate (2.77 g, 19.0 mmol) and ethyl benzyloxyacetate (3.69 g, 19.0 mmol) in dry tetrahydrofuran (30 ml) were treated at 22° C. with sodium hydride (0.83 g of a 60% dispersion in mineral oil, 20.9 mmol) followed by ethanol (10 μl) and the resulting mixture was stirred at 22° C. for 18 h. The tetrahydrofuran was evaporated under reduced pressure to give an orange syrup. A mixture of intermediate 3,2-(2-(methylthio)ethoxy)acetamidine hydrochloride salt, (3.50 g, 19.0 mmol) in a solution of sodium ethoxide (9.5 mmol, prepared from 0.22 g of sodium in ethanol 25 ml) was then added all at once to the above adduct and the resulting mixture heated at 60° C. for 3 h. Acetic acid (2 ml) was added and the ethanol was evaporated under reduced pressure. The residue was diluted with ethyl acetate washed successively with saturated sodium bicarbonate and brine, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Chromatography on silica gel (elution with a gradient of ethyl acetate 20-30% in toluene) gave 0.728 g (10% yield) of the title ester as a clear oil. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.33 (3H, t, J=7.1 Hz, CH3), 2.18 (3H, s, SCH3), 2.78 (2H, t, J=6.0 Hz, CH2), 3.78 (2H, t, J=6.0 Hz, CH2), 4.36 (2H, q, J=7.1 Hz, OCH2), 4.54 (2H, s, OCH2), 5.35 (2H, s, OCH2), 7.37 (3H, m, aromatics), 7.48 (2H, m, aromatics). HRMS (ESI+) calculated for C18H23N2O5S [M+H+]: 379.1328: found: 379.1314.
Intermediate 5




Ethyl 5-benzyloxy-2-{(2-(dimethylsulfonium)ethoxy)methyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate iodide. A solution of intermediate 4, ethyl 5-benzyloxy-2-{(2-(methylthio)ethoxy)methyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate, (0.555 g, 1.47 mmol) in dichloromethane (10 ml) was treated at 22° C. with iodomethane (2.0 ml, 21.5 mmol) for 10 days. Evaporation of the solvent and excess reagent gave the title compound (0.76 g) as an oil which was used without further purification. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.32 (3H, t, J=7.1 Hz, CH3), 3.26 (6H, s, SCH3), 4.02 (2H, m, CH2), 4.23 (2H, m, CH2), 4.34 (2H, q, J=7.1 Hz, OCH2), 4.69 (2H, s, OCH2), 5.23 (2H, s, OCH2), 7.35-7.5 (5H, m, aromatics). MS (ESI+) m/z 393 [M+].
Intermediate 6




Ethyl 3-benzyloxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. A solution of intermediate 5, ethyl 5-benzyloxy-2-{(2-(dimethylsulfonium)ethoxy)methyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate iodide, (0.76 g, 1.47 mmol) in dry N,N-dimethylformamide (10 ml) was treated at 22° C. with powdered anhydrous potassium carbonate (2.5 g) and the resulting mixture stirred for 48 h. The solid was then filtered and the filtrate evaporated in vacuo. The residue was diluted with ethyl acetate washed successively with 0.1 N hydrochloric acid, saturated sodium bicarbonate, brine and dried over anhydrous magnesium sulfate. Evaporation of the solvent followed by chromatography on silica gel (elution with a gradient of ethyl acetate 20-50% in toluene) gave 0.347 g (72% yield) of the title ester as white prisms; mp 103-104° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ ppm: 1.34 (3H, t, J=7.1 Hz, CH3), 4.03 (2H, t, J=5.6 Hz, CH2), 4.11 (2H, t, J=5.6 Hz, CH2), 4.37 (2H, q, J=7.1 Hz, OCH2), 4.74 (2H, s, OCH2), 5.30 (2H, s, OCH2), 7.38 (3H, m, aromatics), 7.50 (2H, m, aromatics). Anal. Calcd for C17H18N2O5: C, 61.81; H, 5.49; N, 8.48. Found: C, 61.55; H, 5.53; N, 8.39.
Intermediate 7




3-(Benzyloxy)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid. A solution of intermediate 6, ethyl 3-(benzyloxy)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate (0.300 g, 0.91 mmol) in ethanol (10 ml) was treated with 3 ml (3.0 mmol) of 1 N sodium hydroxide and stirred at 25° C. for 30 min. The solution was then acidified with 1 N hydrochloric acid, extracted with ethyl acetate, washed with brine and dried over anhydrous magnesium sulfate. Evaporation of the solvent gave 0.264 g (96% yield) of the title acid as white crystals; mp 171° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ (ppm): 4.03 (2H, t, J=5.3 Hz, CH2), 4.12 (2H, t, J=5.3 Hz, CH2), 4.73 (2H, s, OCH2), 5.53 (2H, s, OCH2), 7.35-7.42 (3H, m, aromatics), 7.53 (2H, m, aromatics). Anal. Calcd for C15H14N2O5: C, 59.60; H, 4.67; N, 9.27. Found: C, 59.35; H, 4.69; N, 9.10.
Intermediate 8




Ethyl 3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. A solution of intermediate 6, ethyl 3-benzyloxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.236 g, 0.714 mmol) in a mixture of ethyl acetate (60 ml) and ethanol (20 ml) was treated with 1 atm of hydrogen at 25° C. over 10% palladium on activated carbon (0.10 g) for 2.5 h to give 0.160 g (94% yield) of the title compound as white needles; mp 172-174° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ ppm: 1.47 (3H, t, J=7.3 Hz, CH3), 4.08 (4H, m, 2×CH2), 4.54 (2H, q, J=7.3 Hz, OCH2), 4.72 (2H, s, OCH2), 10.75 (1H, s, OH). Anal. Calcd for C10H12N2O5: C, 50.00; H, 5.03; N, 11.66. Found: C, 50.01; H, 4.95; N, 11.54.
Intermediate 9




2-(2-(Methylthio)ethoxy)propanoic acid. Addition of 2-methylthioethanol (10.0 g, 0.108 mol) to sodium hydride (9.54 g of a 60% dispersion in mineral oil, 0.238 mol, washed twice with hexane) followed by reaction with 2-bromopropionic acid (16.6 g, 0.108 mol) gave 13.81 g (78% yield) of the title compound as a clear oil; bp 80-90° C./0.2 torr (bulb to bulb distillation, air bath temperature). 1HNMR 400 MHz (CDCl3) δ ppm: 1.49 (3H, d, J=7.0 Hz, CH3), 2.18 (3H, s, SCH3), 2.76 (2H, t, J=6.6 Hz, CH2), 3.74 (2H, t, J=6.6 Hz, CH2), 4.07 (1H, d, J=7.0 Hz, OCH).
Intermediate 10




Methyl 2-(2-(methylthio)ethoxy)propanoate. Reaction of intermediate 9,2-(2-(methylthio)ethoxy)propanoic acid, (13.70 g, 0.083 mol) with oxalyl chloride followed by reaction with methanol gave 14.27 g (96% yield) of the title ester as a clear oil; bp 55-60° C./0.3 torr (bulb to bulb distillation, air bath temperature). 1HNMR 400 MHz (CDCl3) δ ppm: 1.42 (3H, d, J=7.0 Hz, CH3), 2.15 (3H, s, SCH3), 2.71 (2H, t, J=6.8 Hz, CH2), 3.56 (1H, m, CH), 3.75 (3H, s, OCH3), 3.78 (1H, m, CH), 4.15 (1H, q, J=7.0 Hz, OCH).
Intermediate 11




2-(2-(Methylthio)ethoxy)propanamidine hydrochloride salt. Intermediate 10, methyl 2-(2-(methylthio)ethoxy)propanoate, (10.00 g, 56.1 mmol) was added to a solution of methylchloroaluminum amide (0.224 mol; prepared in toluene (100 ml) from ammonium chloride 12.36 g (0.231 mol) and 112.0 ml (0.224 mol) of a 2 M solution of trimethylaluminum in toluene} as described in the preparation of intermediate 3 to give 7.70 g (69% yield) of the title compound as an oil. 1HNMR 400 MHz (D2O) δ ppm: 1.37 (3H, d, J=6.6 Hz, CH3), 2.01 (3H, s, SCH3), 2.65 (2H, t, J=5.6 Hz, CH2), 3.64 (2H, m, CH2), 4.30 (1H, q, J=6.6 Hz, OCH). MS (ESI+) m/z 163 [M+H+].
Intermediate 12




Ethyl 5-benzyloxy-2-{1-(2-(methylthio)ethoxy)ethyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate. Diethyl oxalate (5.66 g, 38.7 mmol) and ethyl benzyloxyacetate (7.52 g, 38.7 mmol) in dry tetrahydrofuran (60 ml) were treated at 22° C. with sodium hydride (1.70 g of a 60% dispersion in mineral oil, 42.5 mmol) and the condensation product was reacted with a mixture of intermediate 11, 2-(2-(methylthio)ethoxy)propanamidine hydrochloride salt, (7.70 g, 38.7 mmol) in a solution of sodium ethoxide (19.3 mmol, prepared from 0.445 g of sodium) in ethanol (50 ml) to give 2.29 g (15% yield) of the title ester as a clear oil after chromatography on silica gel. 1HNMR 400 MHz (CDCl3) δ ppm: 1.33 (3H, t, J=7.1 Hz, CH3), 1.54 (3H, d, J=7.1 Hz, CH3), 2.16 (3H, s, SCH3), 2.7-2.8 (2H, m, CH2), 3.54 (1H, m, CH), 3.86 (1H, m, CH), 4.37 (2H, q, J=7.1 Hz, OCH2), 4.47 (1H, q, J=7.1 Hz, OCH), 5.34 (2H, ABq, JAB=11.0 Hz, OCH2), 7.37 (3H, m, aromatics), 7.49 (2H, m, aromatics). MS (ESI+) m/z 393 [M+H+].
Intermediate 13




Ethyl 5-benzyloxy-2-{1-(2-(dimethylsulfonium)ethoxy)ethyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate iodide. A solution of intermediate 12, ethyl 5-benzyloxy-2-{1-(2-(methylthio)ethoxy)ethyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate, (1.63 g, 4.15 mmol) in dichloromethane (5 ml) was treated at 22° C. with iodomethane (5.0 ml, 53.9 mmol) for 5 days as described for the preparation of intermediate 5, to give the title compound (2.22 g) as an oil which was used without further purification. 1HNMR 400 MHz (CDCl3) δ ppm: 1.32 (3H, t, J=7.1 Hz, CH3), 1.66 (3H, d, J=6.6 Hz, CH3), 3.15 (3H, s, SCH3), 3.32 (3H, s, SCH3), 3.4 (1H, m, CH), 4.01 (2H, m, CH2), 4.37 (2H, q, J=7.1 Hz, OCH2), 4.45 (1H, m, CH), 4.63 (1H, q, J=6.6 Hz, OCH), 5.28 (2H, OCH2), 7.38 (3H, m, aromatics), 7.50 (2H, m, aromatics). MS (ESI+) m/z 407 [M+].
Intermediate 14




Ethyl 3-(benzyloxy)-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. A solution of intermediate 13, ethyl 5-benzyloxy-2-{1-(2-(dimethylsulfonium)ethoxy)ethyl}-6-oxo-1,6-dihydropyrimidine-4-carboxylate iodide (2.22 g, 4.15 mmol) in dry N,N-dimethylformamide (30 ml) was treated at 22° C. with powdered anhydrous potassium carbonate (6 g) and stirred for 40 h. The solid was then filtered and the filtrate was concentrated in vacuo. The residue was diluted with ethyl acetate, washed with 0.1 N hydrochloric acid, saturated sodium bicarbonate and brine then dried over anhydrous magnesium sulfate. Evaporation of the solvent and chromatography of the residue on silica gel (elution toluene-ethyl acetate 7:3) gave 1.0 g (70% yield) of the title ester as white crystals; mp 48-50° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.33 (3H, t, J=7.1 Hz, CH3), 1.68 (3H, d, J=6.6 Hz, CH3), 3.91 (2H, m, CH2), 4.17-4.31 (2H, m, CH2), 4.37 (2H, q, J=7.1 Hz, OCH2), 4.73 (1H, q, J=6.6 Hz, OCH), 5.30 (2H, ABq, JAB=11.0 Hz, OCH2), 7.38 (3H, m, aromatics), 7.50 (2H, m, aromatics). Anal. Calcd for C18H20N2O5: C, 62.78; H, 5.85; N, 8.13. Found: C, 62.69; H, 6.01; N, 8.16.
Intermediate 15




Ethyl 3-hydroxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. Hydrogenolysis of intermediate 14, ethyl 3-benzyloxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.610 g, 1.77 mmol) in a mixture of ethyl acetate (75 ml) and ethanol (75 ml) at 25° C. over 10% palladium on activated carbon (0.20 g) under 1 atm of hydrogen for 2 h gave 0.430 g (95% yield) of the title ester as white crystals; mp 119-121° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ ppm: 1.46 (3H, t, J=7.1 Hz, CH3), 1.67 (3H, d, J=6.6 Hz, CH3), 3.90 (2H, m, CH2), 4.13-4.32 (2H, m, CH2), 4.51 (2H, m, OCH2), 4.70 (1H, q, J=6.6 Hz, CH), 10.7 (1H, broad, OH). Anal. Calcd for C11H14N2O5: C, 51.96; H, 5.55; N, 11.01. Found: C, 51.60; H, 5.61; N, 10.70.
Intermediate 16




3-Benzyloxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid. Saponification of intermediate 14, ethyl 3-benzyloxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.225 g, 0.65 mmol) as described in the preparation of intermediate 7 gave 0.198 g (96% yield) of the title acid as white crystals; mp 167-168° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ ppm: 1.68 (3H, d, J=6.6 Hz, CH3), 3.93 (2H, m, CH2), 4.12-4.21 (1H, m, CH), 4.27-4.35 (1H, m, CH), 4.70 (1H, q, J=6.6 Hz, OCH), 5.53 (2H, ABq, JAB=11.1 Hz, OCH2), 7.39 (3H, m, aromatics), 7.55 (2H, m, aromatics). MS (ESI+) m/z 317 [M+H+]. Anal. Calcd for C1-6H16N2O5: C, 60.75; H, 5.10; N, 8.86. Found: C, 60.65; H, 5.05; N, 8.72.
Intermediate 17




2-(2-(Methylthio)ethyl)-1,3-dioxolane. A solution of 3-(methylthio)propanal (5.2 g, 0.05 mol) and ethyleneglycol (3.4 g. 0.055 mol) in 100 mL of benzene was treated with 300 mg p-toluenesulfonic acid and heated at reflux for 4 hrs. The solution was cooled and decanted. Concentration and drying in vacuo provided the title compound as a light yellow oil. 1H NMR (300 MHz, CDCl3) δ ppm:4.93 (1H, t, J=0.76 Hz) 3.74-4.03 (4H, m) 2.46-2.68 (2H, m) 2.08 (3H, s) 1.83-2.00 (2H, m).
Intermediate 18




4-(Methylthio)-2-(2-(trimethylsilyloxy)ethoxy)butanenitrile. Intermediate 17, 2-(2-(methylthio)ethyl)-1,3-dioxolane, (2.96 g, 0.02 mol), trimethylsilylcyanide (1.98 g, 0.02 mol) and 20 mg zinc iodide were combined under N2 and stirred for 16 hrs at room temperature. The mixture was then concentrated in vacuo to provide 4.9 g (approximately 100% yield) of the title compound as a yellow oil. 1H NMR (300 MHz, CDCl3) δ ppm:4.37-4.49 (1H, m) 3.50-3.88 (4H, m) 2.57-2.74 (2H, m) 1.98-2.26 (5H, m) 0.06-0.22 (9H, m): LC/MS198 (-TMS+Na).
Intermediate 19




4-(Methylthio)-2-(2-(trimethylsilyloxy)ethoxy)butanamidine. A solution of intermediate 18, 4-(methylthio)-2-(2-(trimethylsilyloxy)ethoxy)butanenitrile, (4.9 g, 0.02 mol) in 30 mL of methanol was saturated with ammonia. The flask was then sealed and heated in an oil bath at 80-90° C. for 16 hrs. After cooling, the flask was opened and the mixture concentrated in vacuo to give the title compound in essentially quantitative yield as a very viscous oil. 1H NMR (300 MHz, CDCl3) δ ppm:3.94-4.02 (1H, m) 3.76-3.94 (3H, m) 3.68-3.77 (2H, m) 3.52-3.62 (2H, m) 2.53-2.67 (2H, m) 2.07 (3H, s) 1.89-2.01 (2H, m); LC/MS193 (M+H).
Intermediate 20




Ethyl 5-(benzyloxy)-2-(1-(2-hydroxyethoxy)-3-(methylthio)propyl)-6-oxo-1,6-dihydropyrimidine-4-carboxylate. Ethyl 2-(benzyloxy)acetate (7.76 g, 0.04 mole) and diethyloxalate (5.84 g, 0.04 mole) in 80 mL of tetrahydrofuran were treated with one equivalent of NaH and a few drops of ethanol. The resulting mixture was stirred for 1.5 hours after which the solvent was removed under vacuum and replaced with 30 mL of ethanol. Intermediate 19, 2-(2-hydroxyethoxy)-4 methylthio)butanamidine, in 30 mL ethanol was added to the mixture followed by NaH (60% in mineral oil, 800 mg, 0.02 mol). This was stirred for 20 hrs at room temperature and 3 hrs at 60° C. then concentrated under reduced pressure. The residue was dissolved in CH2Cl2 and washed with water. The CH2Cl2 layer was dried over MgSO4, filtered and concentrated under vacuum. Chromatography on silica gel, eluting with 4:1 CH2Cl2; ether and ethyl acetate, gave 760 mg of the title compound (9% yield). 1H NMR (300 MHz, CDCl3) δ ppm: 7.24-7.54 (5H, m) 5.17-5.36 (2H, s) 4.50 (1H, m) 4.30 (2H, q, J=7.32 Hz) 3.38-4.00 (4H, m) 2.59 (2H, m) 1.95-2.11 (5H, m) 1.20-1.36 (3H, t, J=7.32 Hz); LC/MS m/z 423 (M+H).
Intermediate 21




Ethyl 3-(benzyloxy)-9-(2-(methylthio)ethyl)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. To a solution of intermediate 20, ethyl 5-(benzyloxy)-2-(1-(2-hydroxyethoxy)-3-(methylthio)propyl)-6-oxo-1,6-dihydropyrimidine-4-carboxylate, (527 mg, 1.25 mmol) and Et3N (505 mg, 5 mmol) dissolved in 10 mL of CH2Cl2 was added a solution of CH3SO2Cl (288 mg, 2.5 mmol) dissolved in 2 mL of CH2Cl2. This was stirred for 20 hrs then concentrated. The crude product was purified by chromatography on silica gel, using 10:1 CH2Cl2:ether as eluent, to give the title compound 265 mg (52% yield). (500 MHz, CDCl3) δ ppm: 7.31-7.55 (5H, m) 5.30 (2H, s) 4.75 (1H, dd, J=3.66 Hz) 4.32-4.40 (2H, q, J=7.17 Hz) 4.13-4.30 (2H, m) 3.75-3.97 (2H, m) 2.20-2.84 (4H, m) 2.06 (3H, s) 1.32 (3H, t, J=7.17 Hz); LC/MS m/z 405 (M+H).
Intermediate 22




3-(Benzyloxy)-9-(2-(methylthio)ethyl)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid. To a stirred solution of intermediate 21, ethyl 3-(benzyloxy)-9-(2-(methylthio)ethyl)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate (97 mg, 0.2 mmol) in 3 mL tetrahydrofuran was added lithium hydroxide (15 mg, 0.6 mmol) in 3 mL water. After 20 min the reaction mixture was acidified with 1N HCl and extracted with CH2Cl2. The extract was dried over MgSO4, filtered and concentrated to give 82 mg of the title compound (88% yield). LC/MS m/e 377.
Intermediate 23




2-(2-Chloroethoxy)-2-methylpropanenitrile. (Navalokina, R. Et al J. Org. Chem. USSR (Engl. Trans.), 1980, 16, 1382-1386.2) Ramalingam, K. U.S. Pat. No. 4,864,051, 1989.). A 250 mL round bottom flask was charged with ZnCl2 (68.14 g, 0.5 mole) which was then fused by heating under vacuum. After returning to room temperature the material was placed under an atmosphere of N2. To this was added acetone cyanohydrin (45.66 mL, 0.5 mole) followed by 2-chloroethanol (50.24 mL, 0.75 mole) and the mixture placed in a preheated oil bath (60° C.). After stirring for 18-20 h at 60° C., the reaction mixture was cooled, diluted with water (300 mL) and washed with CH2Cl2 (5×100 mL). The combined CH2Cl2 extracts were dried (Na2SO4), filtered and concentrated under vacuum to afford the crude product as a yellow liquid. Purification was accomplished by vacuum distillation (10 mm Hg) using a vigreux column. The fraction boiling between 65-75° C. was collected to afford the desired product as a colorless oil (47.1 g, 63.8% yield). 1H NMR (500 MHz, CDCl3) δ ppm: 3.85 (2H, t, J=5.8 Hz), 3.64 (2H, t, J=5.8 Hz), 1.60 (6H, s).
Additional procedure. ZnCl2 (352.3 g 2.59 moles) was added to a 2 L round bottom flask equipped with a mechanical stirrer, nitrogen inlet-outlet, temperature probe, and condenser. Acetone cyanohydrin (110.0 g 1.29 moles) was slowly added to the stirred solid over 30 minutes keeping the temperature below 32° C. with external cooling. To the slurry, 2-chloroethanol (124.9 g 1.55 moles) was slowly added over 20 minutes keeping the temperature below 32° C. with external cooling. Acetone (3.75 g, 64.6 mmoles) was added and the mixture was heated to 60° C. After stirring for 4 h at 60° C., the reaction mixture was cooled to 30-35° C., diluted with water (1.10 L) and extracted with CH2Cl2 (1×440 mL and 1×220 mL). The combined CH2Cl2 extracts were washed with 0.5M sodium bicarbonate (330 mL), followed by water (3×330 mL). The dichloromethane solution was concentrated under vacuum to afford crude product (109 g). The crude product was purified by vacuum distillation (10 mm Hg) using a Vigreux column. The fraction boiling at 60-80° C. was collected to afford 2-(2-chloroethoxy)-2-methylpropanenitrile as colorless oil (88.7 g).
Intermediate 23b
2-(2-Bromoethoxy)-2-methylpropanenitrile. Dichloromethane (42 mL) and tin tetrachloride (64.9 g, 249 mmol) were added to a 250 mL round bottom flask equipped with a magnetic stirrer, temperature probe, condenser and Argon inlet-outlet. The mixture was cooled to 0-5° C. Acetone cyanohydrin (21.2 g, 249 mmol) was added over 15 min, followed by slow addition of 2-bromoethanol (46.69 g, 373.6 mmol). The reaction mixture was stirred at 20-25° C. for 22 h. The mixture was cooled to 0-5° C., diluted with water (148 mL), and extracted with dichloromethane (3×64 mL). The combined CH2Cl2 extracts were dried (MgSO4), filtered and concentrated under vacuum to afford the crude product as a yellow liquid. The material was purified by vacuum distillation (10 mm Hg) using a Vigreux column. The fraction boiling between 75-85° C. was collected to afford 2-(2-Bromoethoxy)-2-methylpropanenitrile as a colorless oil (15.09 g).
Intermediate 24




Ethyl 2-(2-ethoxy-2-oxoethyl)-8,8-dimethyl-2,5,6,8-tetrahydro-[1,2,4]oxadiazolo[3,2-c][1,4]oxazine-2-carboxylate. To a stirred solution of intermediate 23, 2-(2-chloroethoxy)-2-methylpropanenitrile (14.7 g, 0.10 mole) and NaI (1.5 g, 10 mmol) in ethanol (50 mL) was added an aqueous solution (50%) of hydroxylamine (18.4 g, 0.30 mole) resulting in an exothermic reaction. Following this the reaction mixture was heated at 80° C. for 2 h. Upon cooling to room temperature the solvent was removed. The resulting residue was dissolved in 1:1 ethanol/H2O (100 mL) and cooled in an ice bath. To this was added diethyl acetylenedicarboxylate (17.6 mL, 0.110 mole) over 10 min. The reaction mixture was allowed to warm to room temperature and stirred for 1 h. Following this, it was diluted with ethyl acetate (250 mL), washed with H2O (2×100 mL), brine (50 mL), dried over Na2SO4, filtered and concentrated to give the crude product as a yellow oil. Flash chromatography on a silica gel column, eluting with 20-40% ethyl acetate/Hexanes, provided the title compound as a viscous pale yellow oil (15.29 g, 48.6% yield). 1H NMR (500 MHz, CDCl3) δ ppm: 4.35-4.28 (2H, m), 4.18-4.12 (2H, m), 3.60-3.56 (1H, m), 3.51-3.47 (1H, m), 3.30 (1H, d, J=16.2 Hz), 2.94 (1H, d, J=16.2 Hz), 1.52 (3H, s), 1.51 (3H, s), 1.29 (3H, t, J=7.0 Hz), 1.24 (3H, t, J=7.0 Hz). LCMS (M+H) calcd for C14H23N2O7: 315.16; found: 315.33.
Additional procedures. A solution of 2-(2-chloroethoxy)-2-methylpropanenitrile (5.0 g, 33.87 mmol) in anhydrous methanol (33 mL) was added to a 3-necked flask equipped with a reflux condenser and argon inlet-outlet. Hydroxylamine hydrochloride (2.82 g, 40.58 mmol, 1.20 eq) was added, followed by powdered sodium carbonate (3.95 g, 37.26 mmol, 1.10 eq). The resulting suspension was allowed to stir at room temperature under argon for 18 hrs. The reaction mixture was then heated at 70° C. in an oil bath for 3 h. The resulting suspension was allowed to cool to room temperature. The mixture was filtered though celite and the filter cake was washed with additional methanol. The filtrate was concentrated at reduced pressure to give a semi-solid material which was suspended in chloroform ( ̃80 mL). After stirring for 1 hour, the resulting suspension was filtered through celite and the filter cake was washed with additional chloroform. The filtrate was concentrated at reduced pressure to give very pale amber oil which crystallized upon standing to crude cyclic amine oxide (5.79 g).
The above crude intermediate (3.0 g) was dissolved in hot methanol (3 mL). The resulting solution was diluted with ethyl acetate (30 mL). The solution was heated at reflux to distill out the bulk of the methanol. Heating was stopped when the distillate reached 75° C. The resulting solution was allowed to stand 30 min at room temperature and 2.5 h at 5° C. The crystalline solid was collected by filtration, washed with ethyl acetate, and dried in vacuo over night at 45-50° C. to give an anhydrous form the cyclic amine oxide as a colorless crystalline solid, 1.94 g.
The crude intermediate above (43.9 g) prepared from a similar experiment described above was dissolved in hot methanol (45 mL). The resulting solution was diluted with ethyl acetate (400 mL) and water (11.5 mL). The solution was heated at reflux to distill out the bulk of the methanol. Heating was stopped when the distillate reached 71.5° C. The resulting solution was diluted with 100 mL of ethyl acetate and 1 mL of methanol (needed to prevent oiling), seeded with a sample of crystalline monohydrate and let slowly cool to room temperature over night. The solid was collected by filtration, washed with ethyl acetate (100 mL), dried first under nitrogen and in vacuo at room temperature to give the monohydrate form the intermediate cyclic amine oxide as a colorless crystalline solid, 27.93 g.
Ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. ZnCl2 (544.5 g 4.0 moles) was added to a 3 L round bottom flask equipped with a mechanical stirrer, nitrogen inlet-outlet, temperature probe, and condenser. Acetone cyanohydrin (170.0 g, 2.00 mol) was slowly added to the stirred solid over 30 minutes keeping the temperature below 32° C. with external cooling. To the slurry, 2-chloroethanol (193.0 g, 2.40 mol) was slowly added over 20 minutes keeping the temperature below 32° C. with external cooling. Acetone (5.80 g, 99.8 mmol) was added and the mixture was heated to 60° C. After stirring for 4 h at 60° C., the reaction mixture was cooled to 30-35° C., diluted with water (1.70 L) and extracted with CH2Cl2 (1×680 mL and 1×340 mL). The combined CH2Cl2 extracts were washed with 0.5M sodium bicarbonate (510 mL), followed by water (3×510 mL). The dichloromethane solution (1210 mL) contained 152 g of 2-(2-chloroethoxy)-2-methylpropanenitrile by GC quantification, GC purity: 94.7%.
The above dichloromethane solution of 2-(2-chloroethoxy)-2-methylpropanenitrile (600 mL, 76 g, 0.515 moles) was added to a 1 L round bottom flask equipped with a mechanical stirrer, nitrogen inlet, temperature probe, and condenser. The solution was concentrated at atmospheric pressure and CH2Cl2 was replaced by methanol (450 mL). The methanol solution was cooled to 20-25° C. and sodium carbonate (21.83 g, 0.206 mmol) was added. An aqueous solution (50%) of hydroxylamine (33.65 mL, 0.617 mol) was added, followed by water (76 mL). The thin slurry was stirred at 20-25° C. for 18 h. Then the reaction mixture was heated to reflux (temperature about 65° C.) for 3 h. HPLC analysis showed the solution contained 61.5 g of 2,2-dimethyl-4-oxy-5,6-dihydro-2H-1,4-oxazin-3-ylamine; with HPLC purity of 97.2.
The above solution of cyclic amine oxide was cooled to 20-25° C. and the pH (7.0) was adjusted to 7.5 with 0.5M sodium carbonate (20 mL). The mixture was cooled to 0 to −5° C. Diethyl acetylenedicarboxylate (78.85 g, 0.463 moles) was added over 1 h keeping the temperature below 5° C. The mixture was stirred for 30 min. Ethyl acetate (760 mL) and water (380 mL) were added. The phases were separated and the aqueous phase was extracted with ethyl acetate (380 mL). The two ethyl acetate solutions were combined, washed twice with 50% brine (each 380 mL). The ethyl acetate solution contained ethyl 2-(2-ethoxy-2-oxoethyl)-8,8-dimethyl-2,5,6,8-tetrahydro-[1,2,4]oxadiazolo[3,2-c][1,4]oxazine-2-carboxylate. Estimated amount of the titled intermediate in the solution by HPLC was 130 g, HPLC AP 79.0. A sample was purified by reverse-phase chromatography giving an oil of 99% purity by HPLC.
Alternate procedure. To a solution of 2-(2-chloroethoxy)-2-methylpropanenitrile (5.00 g, 33.87 mmol) in methanol (30 mL) was added at 20-22° C. a 50% aqueous solution of hydroxylamine (2.21 mL, 40.58 mmol). Sodium carbonate (1.44 g, 13.59 mmol) and finally water (5 mL) were added. The suspension was stirred at 20-25° C. for 18 h. The reaction mixture was heated at reflux (65-66° C.) for 3 h. The resulting solution was cooled to 20-25° C. The pH (7.5) required no adjustment. The mixture was cooled to −5° C. and dimethyl acetylenedicarboxylate (3.75 mL, 30.56 mmoles) was added slowly over about 18 min keeping the temperature between −5° C. to 0° C. The mixture was stirred for 60 min. Additional dimethyl acetylenedicarboxylate (0.4 mL, 3.26 mmoles) was added and mixture was stirred another 10 min at 0° C. to complete the reaction as confirmed by HPLC. Ethyl acetate (50 mL) and water (25 mL) were added to the reaction mixture. The phases were separated and the aqueous phase was extracted again with ethyl acetate (25 mL). The combined ethyl acetate extracts were washed with about 14% aq. NaCl (2×25 mL). The solvent was removed under vacuum to afford methyl 2-(2-methoxy-2-oxoethyl)-8,8-dimethyl-2,5,6,8-tetrahydro-[1,2,4]oxadiazolo[3,2-c][1,4]oxazine-2-carboxylate as a crude oil (9.4 g, 96.9% yield) with a purity of 71% by HPLC. A sample (0.80 g) was purified by reverse-phase chromatography giving an oil (0.48 g) of 97% purity by HPLC, which crystallized on standing; mp 68-69° C.
Intermediate 25




Ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. A solution of intermediate 24, ethyl 2-(2-ethoxy-2-oxoethyl)-8,8-dimethyl-2,5,6,8-tetrahydro-[1,2,4]oxadiazolo[3,2-c][1,4]oxazine-2-carboxylate (31.16 g) in 1,2,4-trimethylbenzene (200 mL) was heated at 180° C. for 5 h. The resulting dark reaction solution was cooled then concentrated to give a dark brown paste which was taken up into ethyl acetate (250 mL) and extracted with 0.5 M aq Na2CO3 (4×50 mL). The organic layer was discarded and the aqueous layer acidified by carefully adding conc. HCl (20 mL) before being extracted with CH2Cl2 (4×50 mL). The combined CH2Cl2 layers were dried (Na2SO4), filtered and concentrated to give a dark paste which was dissolved in ether (100 mL) and allowed to stand at room temperature in a open flask. The brown/light yellow solid that formed was filtered to afford the title compound. The mother liquor that contained product was re-processed to afford additional material (combined yield  ̃18-20% over two steps). 1H NMR (500 MHz, CDCl3) δ: 10.55 (1H, s), 4.45 (2H, q, J=7.0 Hz), 4.02 (4H, s), 1.61 (6H, s), 1.43 (3H, t, J=7.0 Hz). HRMS (M+H) calcd for C12H17N2O5: 269.1138; found: 269.1149. Anal calcd for C12H16N2O5: C, 53.72; H, 6.01; N, 10.44. Found: C, 53.71; H, 6.04; N, 10.30.
An ethyl acetate solution of the intermediate 24 was placed in a 3 L 3-necked flask equipped with a Dean Stark water separator, stirrer, and temperature probe. Ethyl acetate was removed by distillation and replaced with 1,2,4-trimethylbenzene (1.14 L). The resulting solution was heated at 155° C. for 9 h. The dark reaction mixture was cooled to 20-25° C., diluted with water (760 mL) and extracted twice with 0.5M Na2CO3 (each 760 mL) the organic layer was discarded. The aqueous phases were combined and washed with CH2Cl2 (610 mL), phases were separated and the organic layer was discarded. To the resulting aqueous solution was added CH2Cl2 (300 mL), and acidified to pH 2.0 with 6M sulfuric acid (130 mL). Phases were separated, and the aqueous phase was extracted with CH2Cl2 (300 mL). The combined CH2Cl2 layers were separated in two equal portions A and B for isolation of intermediate 25.
Portion A isolation without charcoal treatment. The CH2Cl2 solution (315 mL) was concentrated to 80 mL at atmospheric pressure. Isopropanol (160 mL) was added and the solution was concentrated to 140 mL at atmospheric pressure. The solution was cooled slowly with stirring to 20-25° C. The resulting slurry was further cooled to 0-5° C. and stirred for 2 h. The solid was filtered, washed with cold isopropanol (70 mL), dried in vacuo at 40-45° C. to afford ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate as off-white flakes (34.8 g).
Portion B isolation with charcoal treatment. The CH2Cl2 solution (315 mL) was concentrated to 80 mL at atmospheric pressure. Isopropanol (160 mL) was added and the solution was concentrated to 160 mL at atmospheric pressure. Another portion of isopropanol (160 mL) was added followed by charcoal (10 g). The mixture was stirred at reflux temperature (about 82° C.) for 15 min. Charcoal was removed by filtration, the charcoal cake washed with hot (about 80° C.) isopropanol (120 mL) and combined with the filtrate. The combined isopropanol solution was concentrated to 140 mL at atmospheric pressure. The solution was cooled slowly with stirring to 20-25° C. The resulting slurry was further cooled to 0-5° C. and stirred for 2 h. The solid was filtered, washed with cold isopropanol (50 mL), dried in vacuo at 40-45° C. to afford ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate as off-white flakes (31.3 g).
Alternate procedure. 2-(2-Bromoethoxy)-2-methylpropanenitrile (3.0 g, 15.62 mmol) and methanol (21 mL) was added to a 50 mL round bottom flask equipped with a magnetic stirrer, nitrogen inlet-outlet, temperature probe, and condenser. The methanol solution was cooled to 0-5° C. An aqueous solution (50%) of hydroxylamine (2.13 mL, 39.05 mmol) was added. The reaction mixture was allowed to warm to room temperature and stirred for 2 h, and then heated at 75° C. for 1.5 h.
The reaction mixture was cooled to 0-5° C. and the pH (7.0) was adjusted to pH (8) with 1M sodium carbonate (3.0 mL). Diethyl acetylenedicarboxylate (2.92 g, 17.18 mmoles) was added over 25 min keeping the temperature below 10° C. The reaction mixture was allowed to warm to room temperature and stirred for 2 h. Ethyl acetate (45 mL) and water (15 mL) were added. The phases were separated, the ethyl acetate solution was washed with water (15 mL), dried (MgSO4) filtered and concentrated under vacuum to afford a mixture containing ethyl 2-(2-ethoxy-2-oxoethyl)-8,8-dimethyl-2,5,6,8-tetrahydro-[1,2,4]oxadiazolo[3,2-c][1,4]oxazine-2-carboxylate as the major component an oil (4.6 g).
The above intermediate (4.6 g) and 1,2,4-trimethylbenzene (23 mL) were added to a 100 mL 3-necked round bottom flask equipped with a stirrer, and temperature probe. The resulting solution was heated at 155° C. for 8 h. The dark reaction mixture was cooled to 20-25° C., and extracted five times with 0.5M Na2CO3 (each 4.6 mL) the organic layer was discarded. The aqueous phases were combined and acidified with conc. HCl (2.3 mL), and then extracted with CH2Cl2 (5×4.6 mL). The combined CH2Cl2 layers were dried (MgSO4) filtered and concentrated under vacuum to give a dark paste (1.6 g) which was crystallized from methyl-t-butyl ether (3 mL). The light brown solid was filtered to afford ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate (351 mg).
Intermediate 25b
Methyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. A solution of the above crude methyl 2-(2-methoxy-2-oxoethyl)-8,8-dimethyl-2,5,6,8-tetrahydro-[1,2,4]oxadiazolo[3,2-c][1,4]oxazine-2-carboxylate (8.34 g) in 1,2,4-trimethylbenzene (80 mL) was heated at 155° C. for 9 h. The resulting dark mixture was cooled to 20-25° C. and diluted with water (50 mL). The product was extracted into 0.5M Na2CO3 (2×50 mL). The organic layer was discarded. The aqueous phases were combined and washed with CH2Cl2 (40 mL). The organic wash was discarded. The aqueous solution was acidified to pH 2.0 with 6M sulfuric acid (9.0 mL) and the product extracted into CH2Cl2 (2×20 mL). The combined CH2Cl2 layers were evaporated in vacuo. The residue was redissolved in isopropanol (75 mL) at 75° C. and the solution was treated with activated charcoal (0.85 g) at 75-80° C. for 20 min. The charcoal was removed by filtration and washed with hot isopropanol. The combined filtrate and wash was concentrated in vacuo to 40 mL. The resulting slurry was cooled slowly with stirring to 10° C. and stirred for 1 h. The solid was filtered, washed with cold isopropanol (10 mL) and dried in vacuo at 40-45° C. to afford methyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate as an off-white crystalline solid (3.68 g).
Intermediate 26




Ethyl 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. To a stirred solution of intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (2.68 g, 10 mmol) and benzyl bromide (1.43 mL, 12 mmol) in DMF (40 mL) was added K2CO3 (2.07 g, 20 mmol). After stirring 48 h at ambient temperature, the reaction mixture was diluted with ether (100 mL), then washed with water (3×30 mL) and brine (20 mL). The organic layer was dried (Na2SO4/activated carbon), filtered and concentrated to give a yellow solid. Trituration with hexanes/ether (9:1) afforded the title compound as an off-white solid (2.79 g, 78% yield). 1H NMR (500 MHz, CDCl3) δ ppm: 7.48-7.45 (2H, m), 7.37-7.30 (3H, m), 5.25 (2H, s), 4.33 (2H, q, J=7.3 Hz), 4.05-3.99 (4H, m), 1.62 (6H, s), 1.29 (3H, t, J=7.3 Hz). HRMS (M+H) calcd for C19H23N2O5: 359.1607; found: 359.1611. Anal calcd for C19H22N2O5: C, 63.67; H, 6.18; N, 7.81; found: C, 63.63; H, 6.16; N, 7.78.
Intermediate 27




3-(Benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid. A mixture of intermediate 26, ethyl 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (2.93 g, 8.2 mmol) and LiOH.H2O (0.84 g, 20 mmol) in 4:1 ethanol/tetrahydrofuran (50 mL) was stirred for 2 h at ambient temperature then concentrated under vacuum. The resulting yellow residue was treated with 1N HCl (25 mL) providing a precipitate that was filtered and dried under vacuum to yield the title compound as a white powder (2.68 g, 99% yield). 1H NMR (500 MHz, CDCl3) δ ppm: 7.54-7.48 (2H, m), 7.37-7.27 (3H, m), 5.44 (2H, s), 4.05-3.93 (4H, m), 1.60 (6H, s).). HRMS (M+H) calcd for C17H19N2O5: 331.1294; found: 331.1308. Anal calcd for C17H18N2O5: C, 61.81; H, 5.49; N, 8.48; found: C, 61.84; H, 5.36; N, 8.25.
Intermediate 28




2-Ethyl-2-hydroxybutanenitrile. To a solution of potassium phosphate monobasic (140 g, 1.11 mole) in water (250 mL) was added 3-pentanone (75.8 g, 0.88 mole), followed by a solution of sodium cyanide (54 g, 1.10 mole) in water (250 mL), and the resulting mixture stirred for 3 hours. The mixture was extracted with diethyl ether (1×250 mL, then 2×100 mL) and the combined ether layers washed with 1.0 N HCl (200 mL). The ether solution was dried (Na2SO4), filtered, and concentrated in-vacuo. The crude product was purified by vacuum distillation (bp 87° C., 10 mmHg) to give the title compound (72.4 g, 3% yield) as a clear oil. 1H NMR (500 MHz, CDCl3) δ ppm: 2.71 (1H, s), 1.82 (2H, q, J=7.5 Hz), 1.76 (2H, q, J=7.5 Hz), 1.10 (6H, t, J=7.5 Hz). 13C NMR (500 MHz, CDCl3) δ ppm: 121.21, 73.53, 32.81, 8.27.
Intermediate 29




2-(2-Chloroethoxy)-2-ethylbutanenitrile. Zinc chloride (68.1 g, 0.5 mol) was fused under vacuum as described in the procedure for the synthesis of intermediate 23. The molten zinc was cooled and the evacuated flask was flushed with nitrogen. The flask was loaded with intermediate 28, 2-ethyl-2-hydroxybutanenitrile, (40.3 g, 0.5 mol) and 2-chloroethanol (50.5 mL, 0.75 mmol) then stirred at 60° C. for 20 hours. The reaction mixture was diluted with water (250 mL) and extracted with dichloromethane (1×250 mL, 4×100 mL). The combined organic layers were dried (sodium sulfate), filtered, and concentrated in-vacuo. The crude product was purified by vacuum distillation (bp 83° C., 10 mmHg) to give the title compound (52 g) containing unreacted intermediate 28. 1H NMR (500 MHz, CDCl3) δ ppm: 3.82 (2H, t, J=5.8 Hz), 3.64 (2H, t, J=5.8 Hz), 1.83 (4H, J=7.3 Hz), 1.03 (6H, t, J=7.6 Hz).
Intermediate 30




Diethyl 2-(2,2-diethyl-3-iminomorpholinooxy)but-2-enedioate. A solution of the product mixture obtained in the synthesis of intermediate 29, 2-(2-chloroethoxy)-2-ethylbutanenitrile, (0.171 mol) in absolute ethanol (150 mL) was added dropwise to a solution of hydroxylamine (50% aqueous solution, 33.8 mL, 0.51 mol), sodium carbonate (9.1 g, 0.086 mol) and sodium iodide (2.55 g, 0.017 mol) over 15 minutes. The mixture was heated at 80° C. for 3 hours. The reaction was then concentrated to a thick paste and azeotroped under vacuum with ethanol/water (1:1, 100 mL), water (100 mL) and finally ethanol (100 mL). The residue was taken up in ethanol/water (1:1, 160 mL), cooled (0° C.), and treated with diethyl acetylenedicarboxylate (30.1 mL, 0.188 mol). The reaction was stirred at room temperature for 2 hours, then diluted with water (200 mL) and ethyl acetate (200 mL). The organic layer was separated, washed with water (200 mL) and brine (100 mL), then dried (sodium sulfate), filtered and concentrated in-vacuo. The crude product was purified by column chromatography over silica gel, eluting with 10% to 40% ethyl acetate in hexanes to afford the title compound (25.7 g) as a yellow oil.
Intermediate 31




Ethyl 9,9-diethyl-3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate. A solution of intermediate 30, diethyl 2-(2,2-diethyl-3-iminomorpholinooxy)but-2-enedioate, (25.7 g) in 1,2,4-trimethylbenzene (100 mL) was heated at reflux (180° C.) for 16 hours. The solvent was then removed in vacuo and the resulting oil placed in a freezer until crystal formation began. The oil-crystal mixture was triturated with diethyl ether (50 mL) and the solid was collected by filtration, washing with a small volume of ether to provide the title compound (9.02 g). A second crop (1.62 g) was obtained from the filtrate. 1H NMR (500 MHz, CDCl3) δ ppm: 10.54 (1H, s), 4.44 (2H, q, J=7.0 Hz), 4.00 (4H, m), 2.00 (2H, m), 1.92 (2H, m), 1.42 (3H, t, J=7.0 Hz), 0.85 (6H, t, J=7.3 Hz). 13C NMR (500 MHz, CDCl3) δ ppm: 169.53, 157.82, 151.40, 147.58, 125.35, 87.27, 62.62, 58.35, 43.24, 31.06, 14.17, 7.79. HRMS [M+H]+ calcd for C14H21N2O5: 297.14506; found: 297.1464.
Intermediate 32




2-(3-Chloropropoxy)-2-methylpropanenitrile. Zinc chloride (68.1 g, 0.5 mol) was fused using the procedure described for the synthesis of intermediate 23, 2-(2-chloroethoxy)-2-methylpropanenitrile. The molten zinc was cooled and the flask flushed with nitrogen. The flask was loaded with acetone cyanohydrin (46 mL, 0.5 mol) and 3-chloropropanol (64 mL, 0.75 mmol) and the reaction mixture stirred at 60° C. for 30 hours. The mixture was then diluted with water (200 mL) and extracted with dichloromethane (1×200 mL and 3×100 mL). The combined organic layers were dried (sodium sulfate), filtered, and concentrated in-vacuo. The crude product was purified by vacuum distillation (bp 78-84° C., 10 mmHg) to give the title compound (41 g) as a 2:1 mixture with residual 3-chloropropanol. 1H NMR (500 MHz, CDCl3) δ ppm: 3.72 (2H, t, J=5.8 Hz), 3.63 (2H, t, J=6.4 Hz), 2.04 (2H, m), 1.57 (6H, br s).
Intermediate 33




Ethyl 2-(2-(3-chloropropoxy)propan-2-yl)-5-hydroxy-6-oxo-1,6-dihydropyrimidine-4-carboxylate. A solution of intermediate 32, 2-(3-chloropropoxy)-2-methylpropanenitrile (0.186 mol) in absolute ethanol (40 mL) was added dropwise to a cold (0° C.) solution of hydroxylamine (50% aqueous solution, 17 mL, 0.278 mol), 20 mL H2O, sodium carbonate (9.91 g, 0.093 mol) and sodium iodide (2.80 g, 0.019 mol) over 15 minutes. (In an alternative procedure, sodium carbonate was omitted from the mixture). The mixture was stirred at room temperature for 30 min, then additional hydroxylamine (17 mL, 0.278 mol) was added. The reaction was then heated at 80° C. for 16 hours. The mixture was concentrated to a thick paste which was azeotroped under vacuum with ethanol/water (1:1, 100 mL). The resulting residue was taken up in ethanol/water (1:1, 200 mL), cooled (0° C.), and treated with diethyl acetylenedicarboxylate (30.1 mL, 0.188 mol) by dropwise addition over 10 min. The reaction was allowed to stir at room temperature for 2.5 hours, then diluted with water (300 mL) and ethyl acetate (300 mL). The separated organic layer was washed with water (100 mL) and brine (100 mL), then dried (sodium sulfate), filtered and concentrated in-vacuo. The crude product was purified by silica gel column chromatography, eluting with 10% to 40% ethyl acetate in hexanes, to give 21.2 g of a yellow oil. A solution of this oil (15.6 g) in 1,2,4-trimethylbenzene (300 mL) was heated at reflux (180° C.) for 2.5 hours after which the solvent was removed in-vacuo. The resulting oil was taken up in ethyl acetate (300 mL) and extracted with saturated aqueous sodium bicarbonate (1×200 mL, then 4×100 mL). The combined aqueous layers were acidified to pH 1-2 using 6 N HCl then extracted with ethyl acetate (3×150 mL). The organic extracts were dried (sodium sulfate), filtered, then concentrated in vacuo. The resulting oil was triturated with diethyl ether (50 mL) and the resulting solid collected by filtration and washed with a small volume of ether to afford the title compound (2.05 g). A second crop (0.70 g) was obtained from the filtrate. 1H NMR (500 MHz, CDCl3) δ ppm: 10.83 (1H, br), 10.02 (1H, br), 4.46 (2H, q, J=7.0 Hz), 3.66 (2H, t, J=6.1 Hz), 3.58 (2H, t, J=5.8 Hz), 2.06 (2H, m), 1.55 (6H, s), 1.44 (3H, t, J=7.0 Hz).
Intermediate 34




Ethyl 3-(benzoyloxy)-10,10-dimethyl-4-oxo-6,7,8,10-tetrahydro-4H-pyrimido[2,1-c][1,4]oxazepine-2-carboxylate. A solution of intermediate 33, ethyl 2-(2-(3-chloropropoxy)propan-2-yl)-5-hydroxy-6-oxo-1,6-dihydropyrimidine-4-carboxylate, (0.064 g, 0.2 mmol) in pyridine (1 mL) was treated with benzoic anhydride (0.047 g, 0.2 mmol) and stirred for 1 hr at 60° C. The solvent was removed and the residue taken up in N,N-dimethylformamide (1 mL) and treated with potassium carbonate (0.036 g, 0.2 mmol). The mixture was stirred for 1 hr at 80° C., and solvent was removed to give the title compound.
Intermediate 35




3-(Benzyloxy)-10,10-dimethyl-4-oxo-6,7,8,10-tetrahydro-4H-pyrimido[2,1-c][1,4]oxazepine-2-carboxylic acid. A suspension of intermediate 33, ethyl 2-(2-(3-chloropropoxy)propan-2-yl)-5-hydroxy-6-oxo-1,6-dihydropyrimidine-4-carboxylate, (0.205 g, 0.64 mmol) and anhydrous potassium carbonate (0.361 g, 2.6 mmol) in anhydrous dimethylformamide (4 mL) was stirred at 60° C. for 5 hours. The reaction mixture was treated with benzyl bromide (0.122 g, 0.71 mmol) and stirred for 16 hours. Following this, 2 mL of H2O was added and the mixture stirred for an additional 24 hours. Solvent was removed by rotary evaporator and the resulting residue suspended in 0.5 N hydrochloric acid (16 mL). The crude product was extracted with ethyl acetate (2×15 mL), then dried (sodium sulfate), filtered, and concentrated to dryness by rotary evaporator to give 0.299 g (Yield>100%) of the title compound as a solid. LC/MS [M+H]+=345.21.
Intermediate 36




3-Hydroxy-10,10-dimethyl-4-oxo-6,7,8,10-tetrahydro-4H-pyrimido[2,1-c][1,4]oxazepine-2-carboxylate. A solution of intermediate 33, ethyl 2-(2-(3-chloropropoxy)propan-2-yl)-5-hydroxy-6-oxo-1,6-dihydropyrimidine-4-carboxylate, (7.01 g, 22 mmol) and anhydrous potassium carbonate (9.12 g, 66 mmol) in anhydrous dimethylformamide (50 mL) was stirred at 80° C. for 20 hours. Solvent was removed by rotary evaporator and the residue, dissolved in water (50 mL), was brought to pH 1 using 6.0 N HCl. The solution was extracted with ethyl acetate (4×25 mL). The combined organic layers were dried (sodium sulfate) and filtered. The solvent was removed by rotary evaporator to give the title compound (5.53 g, Yield 89%) as a brown solid: 1H NMR (500 MHz, CDCl3) δ ppm 10.49 (1H, s), 4.56 (2H, br), 4.43 (2H, q, J=7.2 Hz), 3.69 (2H, t, J=6.4 Hz), 1.93-1.99 (2H, m), 1.61 (6H, s), 1.42 (3H, t, J=7.2 Hz); 13C NMR (126 MHz, CDCl3) δ ppm 169.33, 158.30, 153.39, 148.73, 124.45, 82.85, 62.60, 60.71, 38.79, 27.67, 27.35, 14.15; HRMS (ESI) calcd for C13H19N2O5 (M+H) 283.1294, found 283.1305.
Intermediate 37




(4-Fluoronaphthalen-1-yl)methanamine hydrochloride. A solution of 1-cyano-4-fluoronaphthalene (1.05 g, 6.12 mmol) and 1.5 mL of HCl (aq.) in absolute ethanol (50 mL) was stirred under a hydrogen atmosphere (balloon) with 10% palladium on carbon (0.20 g) for 16 hours. The catalyst was removed by filtration through Celite, and the filtrate concentrated under vacuum. The resulting solid was triturated with ether and collected by filtration to give the title compound (0.575 g, 44% yield) as an off white solid.
Intermediate 38




Methyl 2-(aminomethyl)-5-fluorobenzoate trifluoroacetic acid salt. Methyl 2-((tert-butoxycarbonyl)methyl)-5-fluorobenzoate, prepared according to literature methods, was treated with trifluoroacetic acid to provide the title compound. Yield 100%; 1H NMR (300 MHz, DMSO-d6) δ ppm: 3.89 (3H, s) 4.32 (2H, q, J=5.61 Hz) 7.51-7.71 (2H, m) 7.78 (1H, dd, J=9.33, 2.38 Hz) 8.13 (2H, brs); LC/MS m/z 184 (M+H).
Intermediate 39




2-Aminomethyl-5-fluoro-N-methyl-benzamide trifluoroacetic acid salt. To a solution of tert-butyl 4-fluoro-2-(methylcarbamoyl)benzylcarbamate (7.70 g, 27.3 mmol; prepared from 2-bromo-5-fluorobenzoic acid using literature methods) in CH2Cl2 (100 mL) was added CF3CO2H (25 mL) and the mixture stirred at room temperature for 15 min. This was concentrated in vacuo and the residue triturated with diethyl ether to obtain 8.0 g (Yield 99%) of the title compound as a white powder. 1H NMR (300 MHz, D2O) δ ppm: 2.93 (3H, s) 4.20 (2H, s) 7.35 (1H, dt, J=8.5, 3 Hz) 7.42 (1H, dd, J=9.0, 2.7 Hz) 7.57 (1H, dd, J=8.4, 5.5 Hz); LC/MS m/z 183 (M+H).
Intermediate 40




2-(Aminomethyl)-N-cyclopropyl-5-fluorobenzamide trifluoroacetic acid salt. A solution of tert-butyl 2-(cyclopropylcarbamoyl)-4-fluorobenzylcarbamate (130 mg, 0.42 mmol) prepared according to literature methods, in CH2Cl2 (5 mL) was stirred with trifluoroacetic acid (3 mL) at room temperature for 10 min, then concentrated in vacuo to give 140 mg (Yield 100%) of the title compound as a foam: 1H NMR (DMSO-d6, 300 MHz) δ ppm: 0.62 (2H, m, CH2), 0.73 (2H, m, CH2), 2.86 (1H, m, CH), 4.02-4.07 (2H, ABq, NCH2), 7.46 (2H, m, Ar—Hs), 7.58 (1H, m, Ar—H), 8.11 (3H, br, NH3), 8.81 (1H, d, J=4.4 Hz, NH); LC/MS m/z 209 (M+H).
Intermediate 41




(5-Fluoro-2-methylphenyl)(morpholino)methanone. To a solution of morpholine (870 mg, 10 mmol) and triethylamine (1.1 g, 10.8 mmol) in CH2Cl2 (15 mL) was added a solution of 5-fluoro-2-methylbenzoyl chloride (1.72 g, 10 mmol) in CH2Cl2 (5 mL), dropwise, and the mixture stirred for 15 min. The mixture was then washed with water, and the organic phase dried (MgSO4), filtered, and concentrated to obtain 2.19 g (Yield 98%) of the title compound as a solid: 1H NMR (500 MHz, CDCl3) δ ppm: 2.27 (3H, s) 3.24 (2H, d, J=4 Hz) 3.58 (2H, s) 3.79 (4H, dd, J=18, 3.8 Hz) 6.88 (1H, dd, J=8.2, 2.8 Hz) 6.92-7.05 (1H, m) 7.18 (1H, dd, J=8.4, 5.3 Hz).
Intermediate 42




(2-(Bromomethyl)-5-fluorophenyl)(morpholino)methanone. A mixture of intermediate 41, (5-fluoro-2-methylphenyl)(morpholino)methanone, (2.1 g, 9.5 mmol) and N-bromosuccinimide (2.0 g, 11 mmol) in CCl4 (30 mL) was heated at reflux. To this mixture was added benzoylperoxide (242 mg, 1 mmol) and the mixture heated at reflux for 2 hrs. After cooling, the insoluble materials were filtered and the filtrate purified by column chromatography (SiO2, 0-10% ether in CH2Cl2) to give 1.1 g (Yield 38%) of the title compound as a clear oil: 1H NMR (300 MHz, CDCl3) δ ppm: 3.31 (2H, t, J=4.94 Hz) 3.55-4.02 (6H, m) 4.56 (2H, dd, J=128.81, 9.51 Hz) 6.89 (1H, dd, J=8.23, 2.74 Hz) 6.96-7.12 (1H, m) 7.33-7.49 (1H, m); LC/MS m/z 302 (M+H).
Intermediate 43




(2-(Azidomethyl)-5-fluorophenyl)(morpholino)methanone. To a solution of intermediate 42, (2-(bromomethyl)-5-fluorophenyl)(morpholino)methanone, (1.0 g, 3.32 mmol) in dimethylformamide (10 mL) was added sodium azide (230 mg, 3.5 mmol) and the mixture stirred under a nitrogen atmosphere for 1 h. The solvent was evaporated in vacuo, and the residue dissolved in CH2Cl2, then washed with water. The organic phase was dried (Na2SO4), filtered, concentrated, and the residue purified by column chromatography (SiO2, CH2Cl2) to provide 770 mg (Yield 88%) of the title compound as an oil: 1H NMR (300 MHz, CDCl3) δ ppm: 3.27 (2H, s) 3.51-3.65 (2H, m) 3.66-3.97 (4H, m) 4.38 (2H, brs) 6.92 (1H, dd, J=8.2, 2.7 Hz) 7.07 (1H, dt, J=8.5, 3 Hz) 7.34 (1H, dd, J=8.4, 5.5 Hz); LC/MS m/z 265 (M+H).
Intermediate 44




(2-(Aminomethyl)-5-fluorophenyl)(morpholino)methanone hydrochloride. To a solution of intermediate 43, (2-(azidomethyl)-5-fluorophenyl)(morpholino)methanone, (770 mg, 2.92 mmol,) in ethanol (20 mL) was added 4N HCl (1 mL) and 10% Pd—C (100 mg), and the mixture hydrogenated at 1 atm of H2 for 3 hrs. The catalyst was removed by filtration and the filtrate concentrated. The residue was purified by C18 reverse phase silica gel column chromatography (YMC ODS, 0-5% CH3CN/H2O) to obtain 350 mg (Yield 44%) of the title compound, (2-(aminomethyl)-5-fluorophenyl)(morpholino)-methanone hydrochloride as a white powder: 1H NMR (300 MHz, DMSO-d6) δ ppm: 3.0-4.0 (8H, m), 3.78 (2H, t, J=5 Hz), 7.32 (1H, dd, J=8.8, 2.6 Hz), 7.35-7.44 (1H, t, J=8.5, 3 Hz), 7.75 (1H, dd, J=8.8, 5.5 Hz); LC/MS m/z 239 (M+H).
Intermediate 45




5-Fluoro-2,N,N-trimethyl-benzenesulfonamide. To a solution of 5-fluoro-2-methyl-benzenesulfonyl chloride (4.18 g, 20 mmol) in tetrahydrofuran (25 mL) was added, dropwise, a solution of dimethylamine in tetrahydrofuran (2M, 25 mL, 50 mmol) over 15 min. and the mixture stirred for 5 min. The insoluble materials were filtered and the filtrate concentrated. The residue was purified by column chromatography (SiO2, 5% ether in CH2Cl2) to provide 4.3 g (Yield 90%) of the title compound as a clear oil: 1H NMR (500 MHz, CDCl3) δ ppm: 2.57 (3H, s) 2.82 (3H, s) 2.82 (3H, s) 7.12-7.18 (1H, m) 7.28 (1H, dd, J=8.2, 5.5 Hz) 7.59 (1H, dd, J=8.2, 2.1 Hz); LC/MC m/z 218 (M+H).
Intermediate 46




2-Bromomethyl-5-fluoro-N,N-dimethyl-benzenesulfonamide. Under nitrogen, a mixture of intermediate 45, 5-fluoro-2,N,N-trimethyl-benzenesulfonamide, (435 mg, 2.0 mmol) and N-bromosuccinimide (391 mg, 2.2 mmol) in CCl4 (20 mL) was stirred at 80-90° C. for 5 min. To this mixture was added 2,2′-azobisisobutyronitrile (AIBN, 100 mg) and stirring continued at 80-90° C. for 30 min. After cooling, the insoluble precipitates were filtered and the filtrate concentrated and purified by column chromatography (SiO2, CH2Cl2) to provide 440 mg (Yield 74%) of the title compound; 1H NMR (500 MHz, CDCl3) δ ppm: 2.87 (6H, s) 4.86 (2H, s) 7.28 (1H, dd, J=8.55, 2.75 Hz) 7.61-7.65 (2H, m); LC/MC m/z 296/298 (M+H).
Intermediate 47




2-Azidomethyl-5-fluoro-N,N-dimethyl-benzenesulfonamide. A mixture of intermediate 46, 2-bromomethyl-5-fluoro-N,N-dimethyl-benzenesulfonamide, (880 mg, 2.97 mmol) and sodium azide (200 mg, 3 mmol) in dimethylformamide (4 mL) was stirred at 55-60° C. for 30 min after which the solvent was removed in vacuo. The residue was partitioned between CH2Cl2 and water, and the organic phase was washed with water, dried (Na2SO4), filtered and concentrated to provide 670 mg (Yield 87%) of the title compound as a yellow oil; 1H NMR (500 MHz, CDCl3) δ ppm: 2.84 (6H, s) 4.78 (2H, s) 7.29-7.34 (1H, m) 7.59-7.64 (2H, m).
Intermediate 48




2-(Aminomethyl)-5-fluoro-N,N-dimethylbenzenesulfonamide. To a solution of intermediate 47, 2-azidomethyl-5-fluoro-N,N-dimethyl-benzenesulfonamide, (660 mg, 2.6 mmol) in tetrahydrofuran (10 mL) and water (2 mL) was added triphenylphosphine (740 mg, 2.8 mmol), and the mixture stirred under nitrogen for 1 hr. The tetrahydrofuran was evaporated in vacuo and a mixture of the residue and 6N HCl (3 mL) in MeOH (5 mL) was heated at 80° C. for 20 hrs. This was washed with CH2Cl2, and the aqueous phase basified with dilute NH4OH and extracted with CH2Cl2. The organic extract was dried (Na2SO4), filtered and concentrated to provide 210 mg (0.91 mmol, Yield 35%) of the title compound; 1H NMR (500 MHz, CDCl3) δ ppm: 2.84 (6H, s) 4.10 (2H, s) 7.23-7.29 (1H, m) 7.53-7.60 (2H, m); LC/MS m/z 233 (M+H).
Intermediate 49




5-Fluoro-2,N-dimethyl-benzenesulfonamide. To a solution of 5-fluoro-2-methyl-benzenesulfonyl chloride (4.18 g, 20 mmol) in acetone (20 mL) was added a 40% aqueous solution of methylamine (4.5 mL, 60 mmol) under nitrogen and the mixture stirred for 5 min. Acetone was removed in vacuo and the aqueous residue extracted with CH2Cl2. The CH2Cl2 extract was dried (Na2SO4), filtered, concentrated and the residue purified by column chromatography (SiO2, 10% ether in CH2Cl2) to provide 3.9 g (19.2 mmol, Yield 96%) of the title compound as a white solid; 1H NMR (500 MHz, CDCl3) δ ppm: 2.59 (3H, s), 2.67 (3H, d, J=5.5 Hz), 4.41 (1H, brs), 7.13-7.20 (1H, m), 7.29 (1H, dd, J=8.2, 5.5 Hz), 7.69 (1H, J=8.6, 2.1 Hz); LC/MS m/z 204 (M+H).
Intermediate 50




2-Bromomethyl-5-fluoro-N-methyl-benzenesulfonamide. The title compound can be prepared from intermediate 49, 5-fluoro-2,N-dimethyl-benzenesulfonamide, according to the method described for intermediate 46 and purified by column chromatography (SiO2, 5% ether/CH2Cl2). 1H NMR (500 MHz, CDCl3) δ ppm: 2.64 (3H, d, J=5.19 Hz) 4.91 (1H, d, J=3.66 Hz) 4.98 (2H, s) 7.26-7.30 (1H, m) 7.54 (1H, dd, J=8.6, 5.2 Hz) 7.73 (1H, dd, J=8.4, 2.6 Hz); LC/MS m/z 282/284.
Intermediate 51




2-Azidomethyl-5-fluoro-N-methyl-benzenesulfonamide. The title compound can be prepared from intermediate 50, 2-bromomethyl-5-fluoro-N-methyl-benzenesulfonamide, according to the method described for intermediate 47 and purified by column chromatography (SiO2, 5% ether-CH2Cl2). 1H NMR (500 MHz, CDCl3) δ ppm: 2.65 (3H, d, J=5.19 Hz) 4.81 (2H, s) 4.86 (1H, d, J=4.6 Hz) 7.27-7.33 (1H, m) 7.49 (1H, dd, J=8.2, 5.2 Hz) 7.76 (1H, dd, J=8.2, 2.8 Hz).
Intermediate 52




2-(Aminomethyl)-5-fluoro-N-methylbenzenesulfonamide hydrochloride. To a solution of intermediate 51, 2-azidomethyl-5-fluoro-N-methyl-benzenesulfonamide, (560 mg, 2.3 mmol) in ethanol (10 mL) was added 6N HCl (1 mL) and 10% Pd—C (100 mg) and the mixture hydrogenated with 1 atm of H2 for 14 hrs. The catalyst was removed by filtration through Celite and the filtrate concentrated in vacuo to provide 630 mg (Yield>100%) of the title compound. 1H NMR (500 MHz, DMSO-D6) δ ppm: 4.36 (2H, d, J=5.2 Hz) 7.63-7.70 (2H, m) 7.77-7.83 (1H, m) 8.11 (1H, d, J=4.9 Hz) 8.41 (3H, s); LC/MS m/z 219 (M+H).
Intermediate 53




5-Fluoro-2-methyl-benzenesulfonamide. To a solution of 5-fluoro-2-methyl-benzenesulfonyl chloride (4.18 g, 20 mmol) in acetone (20 mL) was added, dropwise, concentrated NH4OH (3 mL) and the resulting mixture stirred for 5 min. Acetone was removed in vacuo and the precipitates were filtered, washed thoroughly with water and dried in vacuo to provide 3.7 g (Yield 98%) of the title compound as a white solid; 1H NMR (500 MHz, DMSO-D6) δ ppm: 2.55 (3H, s) 7.33-7.40 (1H, m) 7.40-7.46 (1H, m) 7.54 (2H, s) 7.59 (1H, dd, J=9.2, 2.7 Hz); LC/MS m/z 190 (M+H).
Intermediate 54




2-Bromomethyl-5-fluoro-benzenesulfonamide. The title compound can be prepared from intermediate 53, 5-fluoro-2-methyl-benzenesulfonamide, according to the method described for intermediate 46, and purified by column chromatography (SiO2, 5% ether/CH2Cl2). 1H NMR (500 MHz, CDCl3) δ ppm: 5.01 (2H, s) 5.16 (2H, brs) 7.25-7.31 (1H, m) 7.53 (1H, dd, J=8.5, 5.2 Hz) 7.80 (1H, dd, J=8.5, 2.7 Hz). LC/MS m/z 268/270 (M+H).
Intermediate 55




2-Azidomethyl-5-fluoro-N-methyl-benzenesulfonamide. The title compound can be prepared from intermediate 54, 2-bromomethyl-5-fluoro-benzenesulfonamide, according to the method described for the preparation of intermediate 47. 1H NMR (300 MHz, CDCl3) δ ppm: 4.82 (2H, s) 5.18 (2H, s) 7.27 (1H, m) 7.45 (1H, dd, J=8.4, 5.5 Hz) 7.79 (1H, dd, J=8.4, 2.6 Hz). LC/MS m/z 253 (M+Na).
Intermediate 56




2-(Aminomethyl)-5-fluorobenzenesulfonamide hydrochloride. The title compound can be prepared from intermediate 55, 2-azidomethyl-5-fluoro-N-methyl-benzenesulfonamide, according to the method described for the preparation of intermediate 48. 1H NMR (500 MHz, DMSO-D6) δ ppm: 4.05 (2H, s) 5.05 (3H, br) 7.44 (1H, dt, J=8.5, 3 Hz) 7.58 (1H, dd, J=9.2, 2.7 Hz) 7.66 (1H, dd, J=8.5, 5.5 Hz). LC/MS m/z 205 (M+H).
Intermediate 57




5-(2-Bromo-5-fluoro-phenyl)-2-methyl-2H-tetrazole. A mixture of 5-(2-bromo-5-fluoro-phenyl)-1H-tetrazole (1.0 g, 4.12 mmol), iodomethane (1.12 g, 10 mmol) and potassium carbonate (1.5 g) in dimethylformamide (5 mL) was stirred at room temperature for 16 hrs, then concentrated in vacuo. The residue was purified by column chromatography (SiO2, CH2Cl2) to provide 650 mg (Yield 61%) of the title compound as a white powder. 1H NMR (500 MHz, CDCl3) δ ppm: 4.45 (3H, s) 7.03-7.11 (1H, m) 7.63 (1H, dd, J=8.9, 3.1 Hz) 7.69 (1H, dd, J=8.9, 5.5 Hz); 13C NMR (126 MHz, CDCl3)δ ppm: 39.86, 116.28, 118.66, 118.76, 130.13, 135.73, 161.74, 163.53; LC/MS m/z 257/259.
Intermediate 58




4-Fluoro-2-(2-methyl-2H-tetrazol-5-yl)-benzonitrile. A mixture of intermediate 57, 5-(2-bromo-5-fluoro-phenyl)-2-methyl-2H-tetrazole (650 mg, 2.53 mmol) and CuCN (224 mg, 2.5 mmol) in dimethylformamide (4 mL) was placed in a sealed tube and heated at 100-110° C. for 20 hrs. After cooling, the insoluble material was filtered, and the filtrate concentrated in vacuo. The residue was dissolved in CH2Cl2, washed with aq. 4N HCl and dil. NH4OH, then dried (MgSO4), filtered, and concentrated. The residual solid was purified by column chromatography (SiO2, CH2Cl2) to obtain 375 mg (Yield 73%) of the title compound as an off-white solid; 1H NMR (500 MHz, CDCl3) δ ppm: 4.48 (3H, s) 7.29 (1H, dd, J=7.6, 2.8 Hz) 7.85 (1H, dd, J=8.6, 5.2 Hz) 8.00 (1H, dd, J=9.0, 2.6 Hz); LC/MS m/z 204.
Intermediate 59




(4-Fluoro-2-(2-methyl-2H-tetrazol-5-yl)phenyl)methanamine hydrochloride. A solution of intermediate 58, 4-fluoro-2-(2-methyl-2H-tetrazol-5-yl)-benzonitrile, (330 mg, 1.62 mmol) in ethanol (15 mL) was mixed with 6N HCl (1 mL) and 10% Pd—C (200 mg) under nitrogen. The mixture was then stirred under hydrogen (1 atm) for 3 hrs. After removing the catalyst, the filtrate was concentrated in vacuo to provide 360 mg (Yield 91%) of the title compound as an off-white solid; 1H NMR (500 MHz, DMSO-D6) δ ppm: 4.42 (2H, d, J=2.75 Hz) 4.49 (3H, s) 7.48-7.56 (1H, m) 7.78 (1H, dd, J=8.7, 5.7 Hz) 7.86 (1H, dd, J=9.8, 2.8 Hz) 8.45 (3H, s); LC/MS m/z 208.
Intermediate 60




5-(2-Bromo-5-fluoro-phenyl)-1-methyl-2H-tetrazole. A mixture of 5-(2-bromo-5-fluoro-phenyl)-1H-tetrazole (1.0 g, 4.12 mmol), iodomethane (1.12 g, 10 mmol) and potassium carbonate (1.5 g) in dimethylformamide (5 mL) was stirred at room temperature for 16 hrs, then concentrated in vacuo. The residue was purified by column chromatography (SiO2, CH2Cl2) to provide 350 mg (Yield 33%) of the title compound as white crystals. 1H NMR (500 MHz, CDCl3) δ ppm: 4.00 (3H, s) 7.18-7.25 (2H, m) 7.72 (1H, dd, J=8.4, 5.0 Hz); 13C NMR (126 MHz, CDCl3) δ ppm: 34.59, 117.73, 119.58, 120.43, 127.57, 135.11, 153.43, 161.69. LC/MS m/z 257/259.
Intermediate 61




4-Fluoro-2-(1-methyl-2H-tetrazol-5-yl)-benzonitrile. 1H NMR (300 MHz, CDCl3) δ ppm: 4.13 (3H, s) 7.38-7.49 (2H, m) 7.86-7.97 (1H, m); LC/MS m/z 204 (M+H).
Intermediate 62




(4-Fluoro-2-(1-methyl-2H-tetrazol-5-yl)phenyl)methanamine hydrochloride. 1H NMR (500 MHz, DMSO-D6) δ ppm: 4.05 (2H, s) 4.09 (3H, s) 7.58-7.67 (1H, m) 7.77 (1H, dd, J=9.3, 2.6 Hz) 7.87 (1H, dd, J=8.7, 5.7 Hz) 8.38 (3H, s); LC/MS m/z 208.
Intermediate 63




3-m-Tolyl-3-trifluoromethyl-3H-diazirine. To a cold stirring solution of 3-m-tolyl-3-trifluoromethyl-diaziridine (2.0 g, 10 mmol. prepared using the methods described in Doucet-Personeni C. et al., J. Med. Chem., 2001, 44, 3203 and Nassal, M. Liebigs Ann. Chem. 1983, 1510-1523 or in Stromgaard, K et al., J. Med. Chem., 2002, 45, 4038-46) in ethanol (20 mL) was added triethylamine (1.5 g, 15 mmol). To this mixture was added tert-butyl hypochlorite (3.25 g, 30 mmol), and the mixture stirred for 5 min. This mixture was poured into 10% aqueous sodium sulfite (100 mL), and extracted with ether. The ether extract was washed with brine, dried (MgSO4), filtered and concentrated. The residue was purified by column chromatography (SiO2, pentane) to provide 1.6 g (Yield 80%) of the title compound. 1H NMR (300 MHz, CDCl3) δ ppm: 2.33 (3H, s) 6.90-7.03 (2H, m) 7.15-7.31 (2H, m).
Intermediate 64




3-(3-Bromomethyl-phenyl)-3-trifluoromethyl-3H-diazirine. To a solution of intermediate 63, 3-m-tolyl-3-trifluoromethyl-3H-diazirine, (200 mg, 1 mmol) in CCl4 (4 mL) was added N-bromosuccinimide (200 mg, 1.1 mmol, re-crystallized from water), and the stirred mixture heated at 85° C. To this was added AIBN (50 mg) and the mixture heated at reflux for an additional 2.5 hrs. After cooling, the mixture was purified by column chromatography (SiO2, pentane) to provide 150 mg (Yield 54%) of the title compound as a clear oil. 1H NMR (300 MHz, CDCl3) δ ppm: 4.42 (2H, s) 7.10-7.17 (2H, m) 7.31-7.45 (2H, m).
Intermediate 65




2-[3-(3-Trifluoromethyl-diaziridin-3-yl)-benzyl]-isoindole-1,3-dione. A mixture of intermediate 64, 3-(3-bromomethyl-phenyl)-3-trifluoromethyl-3H-diazirine, (140 mg, 0.5 mmol) and potassium phthalimide (95 mg, 0.5 mmol) in dimethylformamide (1.5 mL) was stirred at room temperature for 3 hrs. Dimethylformamide was removed in vacuo. The residue was extracted with CH2Cl2, washed with water, then dried (Na2SO4), filtered, and concentrated. The resulting residue was purified by column chromatography (SiO2, 1:1 CH2Cl2/pentane) to provide 140 mg (Yield 82%) of the title compound as a solid; 1H NMR (300 MHz, CDCl3) δ ppm: 4.80 (2H, s) 7.09-7.21 (2H, m) 7.32 (1H, t, J=7.9 Hz) 7.41-7.49 (2H, m) 7.66-7.71 (2H, m) 7.81-7.85 (2H, m); LC/MS m/z 346 (M+H).
Intermediate 66




(3-(3-(Trifluoromethyl)diaziridin-3-yl)phenyl)methanamine. A stirred solution of intermediate 65, 2-[3-(3-trifluoromethyl-diaziridin-3-yl)-benzyl]-isoindole-1,3-dione, (150 mg, 0.43 mmol) in ethanol (2 mL) was treated with hydrazine hydrate (0.4 mL) at room temperature and the solution stirred for 3.5 hrs. After removing ethanol in vacuo, the residue was partitioned between CH2Cl2 and water. The aqueous phase was acidified with dilute HCl, and washed with CH2Cl2. The aqueous phase was basified with dilute NaOH, and extracted with CH2Cl2. The organic extract was dried (MgSO4), filtered, and concentrated to obtain 50 mg (Yield 54%) of (3-(3 (trifluoromethyl)diaziridin-3-yl)phenyl)methanamine and (3-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)methanamine as a 1:1 mixture; 1H NMR (300 MHz, CDCl3) δ ppm: 3.85 (2H, s) 3.88 (2H, s) 7.08 (2H, s) 7.31-7.40 (4H, m) 7.43-7.50 (1H, m, J=6.2 Hz) 7.54 (1H, s); LC/MS m/z 216 (M+H for diazirine) and 218 (M+H for diaziridine).
Intermediates 67-68
To a solution of 2,4-difluorobenzonitrile (10 g, 72 mmol) dissolved in tetrahydrofuran (20 mL), and dimethylformamide (40 mL) was added 1,2,4-triazole sodium salt (6.3 g, 70 mmol) and the mixture stirred at 90° C. for 3 h after which the mixture was filtered and the solvent removed. The resulting residue was adsorbed onto silica gel and intermediates 67 and 68 separated by flash chromatography, eluting with 0% to 30% ethyl acetate/hexanes.
Intermediate 67




4-Fluoro-2-(1H-1,2,4-triazol-1-yl)benzonitrile. Colorless needles (2.46 g, 18% yield) 1H NMR (500 MHz, CDCl3) δ: 8.89 (1H, s), 8.19 (1H, s), 7.85 (1H, dd, J=8.7, 5.6 Hz), 7.60 (1H, dd, J=8.8, 2.4 Hz), 7.28-7.24 (1H, m). LCMS (M+H) calcd for C9H6N4F: 189.05; found: 189.13.
Intermediate e 68




4-(1H-1,2,4-Triazol-1-yl)-2-fluorobenzonitrile. White solid (0.746 g, 6% yield) 1H NMR (500 MHz, CDCl3) δ: 8.66 (1H, s), 8.15 (1H, s), 7.79 (1H, dd, J=8.5, 6.7 Hz), 7.69 (1H, dd, J=9.5, 1.8 Hz), 7.65-7.63 (1H, m). LCMS (M+H) calcd for C9H6N4F: 189.05; found: 189.13.
Intermediate 69




(4-Fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride). Intermediate 67, 4-fluoro-2-(1H-1,2,4-triazol-1-yl)benzonitrile, (2.46 g, 13.13 mmol) was dissolved in hot ethanol (150 mL). To this was added 1N HCl (15 mL) followed by 10% Pd—C (200 mg). The mixture was treated with H2 at 55 psi for 4 h in a Parr shaker then filtered over Celite and the solvent removed under reduced pressure. The resulting residue was partitioned between ethyl acetate and water. The aqueous phase was separated and lyophilized to afford the title compound as a white powder (2.96 g, 99% yield). 1H NMR (500 MHz, CD3OD) δ ppm: 9.51 (1H, s), 8.63 (1H, s), 7.85 (1H, dd, J=8.5, 5.8 Hz), 7.68 (1H, dd, J=8.8, 2.4 Hz), 7.49 (1H, td, J=8.3, 2.4 Hz), 4.20 (2H, s). LCMS (M+H) calcd for C9H10N4F: 193.08; found: 193.16.
Intermediate 70




(2-Fluoro-4-(1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride. The title compound can be prepared from intermediate 68 according to the method described for the synthesis of intermediate 69. White powder (79% yield). 1H NMR (500 MHz, CD3OD) δ ppm: 9.25 (1H, s), 8.46 (1H, s), 7.80 (1H, dd, J=8.6, 5.8 Hz), 7.64 (1H, dd, J=8.8, 2.4 Hz), 7.44 (1H, dd, J=8.3, 2.6 Hz), 4.17 (2H, s). LCMS (M+H) calcd for C9H10N4F: 193.08; found: 193.16.
Intermediates 71-74
Intermediates 71-74 were prepared using the procedure described for the synthesis of intermediate 67-70.
Intermediate 71




4-Fluoro-2-morpholinobenzonitrile 1H NMR (500 MHz, CDCl3) δ ppm: 7.55 (1H, dd, J=8.5, 6.4 Hz), 6.71 (1H, dd, J=8.1, 2.3 Hz), 6.67 (1H, dd, J=11.0, 2.4 Hz), 3.88 (4H, t, J=4.6 Hz), 3.22 (4H, t, J=4.6 Hz). LCMS (M+H) calcd for C11H12N2OF: 207.09; found: 207.19.
Intermediate 72




4-Morpholino-2-fluorobenzonitrile. 1H NMR (500 MHz, CDCl3) δ ppm: 7.42 (1H, dd, J=8.8, 7.6 Hz), 6.63 (1H, dd, J=8.8, 2.4 Hz), 6.56 (1H, dd, J=12.8, 2.4 Hz), 3.84 (4H, t, J=4.9 Hz), 3.28 (4H, t, J=4.9 Hz). LCMS (M+H) calcd for C11H12N2OF: 207.09; found: 207.19.
Intermediate 73




(4-Fluoro-2-morpholinophenyl)methanamine hydrochloride. 1H NMR (500 MHz, CDCl3) δ ppm: 7.54 (1H, t, J=7.3 Hz), 7.20 (1H, dd, J=10.5, 2.0 Hz), 7.05-7.02 (1H, m), 4.28 (2H, s), 3.93 (4H, bs), 3.03 (4H, bs). LCMS (M+H) calcd for C11H16N2OF: 211.12; found: 211.23.
Intermediate 74




(2-Fluoro-4-morpholinophenyl)methanamine hydrochloride. 1H NMR (500 MHz, CD3OD) δ ppm: 7.73 (1H, t, J=8.2 Hz), 7.62 (1H, d, J=7.6 Hz), 7.58 (1H, d, J=8.2 Hz), 4.26 (2H, s), 4.11 (4H, t, J=4.4 Hz), 3.65 (4H, t, J=4.4 Hz). LCMS (M+H) calcd for C11H16N2OF: 211.12; found: 211.23.
Intermediate 75




4-Fluoro-2-(1,1-dioxo-1λ6-[1,2]thiazinan-2-yl)benzonitrile. To a mixture of 2,4-difluorobenzonitrile (10.0 g, 72 mmol) and 1,1-dioxo-1λ6-[1,2]thiazin-2-ane (8.84 g, 65.4 mmol) in 1:1 tetrahydrofuran/dimethylformamide (40 mL) was added potassium carbonate (9.0 g, 65.4 mmol). The mixture was stirred at 90° C. for 18 h then filtered and concentrated. The residue was purified by flash chromatography (SiO2) eluting with 10%-50% ethyl acetate/hexanes followed by recrystallization from hot ethyl acetate/hexane to give the title compound as white needles (0.537 g, 3% yield). 1H NMR (500 MHz, CD3OD) δ ppm: 7.70 (1H, dd, J=8.8, 5.8 Hz), 7.30 (1H, dd, J=8.8, 2.4 Hz), 7.15-7.12 (1H, m), 3.27 (2H, t, J=5.3 Hz), 3.33 (2H, t, J=6.1 Hz), 2.40-2.35 (2H, m), 2.05-2.01 (2H, m). LCMS (M+H) calcd for C11H16N2OF: 255.06; found: 255.19.
Intermediate 76




(4-Fluoro-2-(1,1-dioxo-1λ6-[1,2]thiazinan-2-yl)phenyl)methanamine hydrochloride. Intermediate 75, 4-fluoro-2-(1,1-dioxo-1λ6-[1,2]thiazinan-2-yl)benzonitrile (1.37 g, 5.4 mmol) was dissolved in ethanol (120 mL). To this was added 1N HCl (20 mL) and a catalytic amount of 10% Pd—C. The mixture was shaken under hydrogen at 55 psi for 4 h then filtered through Celite and concentrated to give the title compound as white solid (1.58 g, 100% yield). 1H-NMR (300 MHz, CD3OD) δ ppm: 7.61 (1H, dd, J=8.4, 6.2 Hz), 7.38 (1H, dd, J=9.3, 2.7 Hz), 7.28 (1H, td, J=8.2, 2.7 Hz), 7.26 (2H, dd, J=21.4, 13.7 Hz), 3.93-3.84 (1H, m), 3.50-3.41 (3H, m), 2.40-2.31 (2H, m), 2.04-1.96 (2H, m). LCMS [M+H]+ calcd for C11H16N2O6FS: 259.087; found: 259.24.
Intermediates 77-78
To a solution of 1H-1,2,3-triazole (3.5 g, 50.7 mmol) in tetrahydrofuran (10 mL) and dimethylformamide (20 mL) was added, portionwise, NaH (1.3 g, 51 mmol, 95%). The mixture was stirred at room temp for 30 min. 2,4-Difluorobenzonitrile (7.6 g, 55 mmol) was added and the mixture stirred at 85° C. for 3 h. The white mixture was concentrated and purified by flash chromatography eluting with 0% to 10% ethyl acetate/hexanes to give intermediates 77 and 78.
Intermediate 77




4-Fluoro-2-1,2,3-triazol-2-yl-benzonitrile. White needles (0.34 g, 3% yield). 1H-NMR (300 MHz, CDCl3) δ ppm: 7.92 (2H, s), 7.88-7.79 (2H, m), 7.19-7.12 (1H, m). LCMS [M+H]+ calcd for C9H6N4F: 189.05; found: 189.12.
Intermediate 78




2-Fluoro-4-1,2,3-triazol-2-yl-benzonitrile. White solid (0.097 g, 1% yield). 1H-NMR (300 MHz, CDCl3) δ ppm: 8.03-7.95 (2H, m), 7.86 (2H, s), 7.74-7.69 (1H, m).
Intermediate 79




4-Fluoro-2-1,2,3-triazol-2-yl-benzylamine hydrochloride. Intermediate 77, 4-fluoro-2-1,2,3-triazol-2-yl-benzonitrile, (0.34 g, 1.8 mmol) was dissolved in ethanol (50 mL). 1N HCl (10 mL) was added along with a catalytic amount of 10%-Pd—C. The mixture was shaken under H2 at 55 psi for 4 h after which it was filtered through Celite and concentrated to give the title compound as the corresponding HCl salt. Yellow solid (0.402 g, 98% yield). 1H-NMR (500 MHz, CD3OD) δ ppm: 8.13 (2H, s), 7.87 (1H, dd, J=4.9, 2.6 Hz), 7.73 (1H, dd, J=4.9, 2.6 Hz), 7.34 (1H, td, J=8.2, 2.7 Hz), 4.35 (2H, s). LCMS [M+H]+ calcd for C9H10N4F: 193.08; found: 193.16.
Intermediate 80




(2-Fluoro-4-(2H-1,2,3-triazol-2-yl)phenyl)methanamine: The title compound can be prepared from intermediate 78, 2-fluoro-4-1,2,3-triazol-2-yl-benzonitrile, according to the procedure provide for intermediate 79. 1H-NMR (300 MHz, CD3OD) δ ppm: 8.05-7.96 (2H, m), 8.00 (2H, s), 7.68 (1H, t, J=8.2 Hz), 4.26 (2H, s). LCMS [M+H]+ calcd for C9H10N4F: 193.08; found: 193.14.
Intermediates 81-84
A solution of 2,4-difluorobenzonitrile (7.07 g, 50.8 mmol) and 3-methyl-1H-1,2,4-triazole (4.22 g, 50.8 mmol) in N,N-dimethylformamide (45 ml) was treated with powdered anhydrous potassium carbonate (10 g) and the resulting mixture stirred at 22° C. for 18 h. The solid was then filtered and the filtrate concentrated in vacuo. The residue was diluted with ethyl acetate, washed with water and brine, then dried over anhydrous magnesium sulfate and concentrated. The resulting mixture was purified by a combination of chromatography on silica gel (elution gradient of ethyl acetate in hexane) and reversed phase silica gel to yield intermediates 81-84.
Intermediate 81




4-Fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)benzonitrile. White crystals (ethyl acetate-hexane); mp 117-118° C. 1HNMR 400 MHz (CDCl3) δ ppm: 2.54 (3H, s, CH3), 7.24 (1H, m, CH), 7.62 (1H, dd, J=2.5 Hz and J=9.1 Hz, CH), 7.84 (1H, dd, J=5.6 Hz and J=8.6 Hz, CH), 8.82 (1H, s, CH). Anal. Calcd for C10H7FN4: C, 59.40; H, 3.49; N, 27.71; Found: C, 59.25; H, 3.32; N, 27.81.
Intermediate 82




4-Fluoro-2-(5-methyl-1H-1,2,4-triazol-1-yl)benzonitrile. White crystals (ethyl acetate-hexane); mp 120-121° C. 1HNMR 400 MHz (CDCl3) δ ppm: 2.56 (3H, s, CH3), 7.30 (1H, dd, J=2.5 Hz and J=8.1 Hz, CH), 7.39 (1H, m, CH), 7.91 (1H, dd, J=5.5 Hz and J=8.6 Hz, CH), 8.06 (1H, s, CH). Anal. Calcd for C10H7FN4: C, 59.40; H, 3.49; N, 27.71; Found: C, 59.35; H, 3.70; N, 27.77.
Intermediate 83




2-Fluoro-4-(3-methyl-1H-1,2,4-triazol-1-yl)benzonitrile. White crystals (ethyl acetate-hexane); mp 133-134° C. 1HNMR 400 MHz (CDCl3) δ ppm: 2.52 (3H, s, CH3), 7.61 (1H, dd, J=2 Hz and J=9.1 Hz, CH), 7.67 (1H, dd, J=2 Hz and J=9.6 Hz, CH), 7.79 (1H, dd, J=6.5 Hz and J=8.6 Hz, CH), 8.56 (1H, s, CH). Anal. Calcd for C10H7FN4: C, 59.40; H, 3.49; N, 27.71; Found: C, 59.42; H, 3.24; N, 28.41.
Intermediate 84




2-Fluoro-4-(5-methyl-1H-1,2,4-triazol-1-yl)benzonitrile. White crystals (ethyl acetate-hexane); mp 89-90° C., 1HNMR 400 MHz (CDCl3) δ ppm: 2.69 (3H, s, CH3), 7.49-7.55 (2H, m, 2×CH), 7.83 (1H, dd, J=6.8 Hz and J=8.8 Hz, CH), 8.00 (1H, s, CH). Anal. Calcd for C10H7FN4: C, 59.40; H, 3.49; N, 27.71; Found: C, 59.17; H, 3.22; N, 28.01.
Intermediate 85




(4-Fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride salt. Hydrogenation of intermediate 81, 4-fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)benzonitrile, (0.680 g, 3.36 mmol) gave 0.720 g (88% yield) of the title hydrochloride salt as a white solid. 1HNMR 400 MHz (DMSO-d6) δ ppm: 2.40 (3H, s, CH3), 4.02 (2H, m, NCH2), 7.50 (1H, m, CH), 7.62 (1H, dd, J=2.8 Hz and J=9.3 Hz, CH), 7.84 (1H, dd, J=6.1 Hz and J=9.1 Hz, CH), 9.00 (1H, s, CH). HRMS (ESI+) calculated for C10H12FN4 [M+H+]: 207.1046; found: 207.1047.
Intermediate 86




(4-Fluoro-2-(5-methyl-1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride salt. Hydrogenation of intermediate 82, 4-fluoro-2-(5-methyl-1H-1,2,4-triazol-1-yl)benzonitrile, (0.244 g, 1.20 mmol) gave 0.290 g (100% yield) of the title hydrochloride salt as a white solid. 1HNMR 400 MHz (DMSO-d6) δ ppm: 2.42 (3H, s, CH3), 3.78 (2H, m, NCH2), 7.58 (1H, m, CH), 7.67 (1H, dd, J=2.8 Hz and J=9.3 Hz, CH), 7.90 (1H, dd, J=6.0 Hz and J=8.6 Hz, CH), 8.22 (1H, s, CH). HRMS (ESI+) calculated for C10H12FN4 [M+H+]: 207.1046; found: 207.1041.
Intermediate 87




(2-Fluoro-4-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride salt. Hydrogenation of intermediate 83, 2-fluoro-4-(3-methyl-1H-1,2,4-triazol-1-yl)benzonitrile, (0.220 g, 1.09 mmol) gave 0.260 g (98% yield) of the title hydrochloride salt as a white solid. 1HNMR 400 MHz (DMSO-d6) δ ppm: 2.38 (3H, s, CH3), 4.09 (2H, m, NCH2), 7.75-7.8 (2H, m, 2×CH), 7.83 (1H, dd, J=2 Hz and J=9 Hz, CH), 9.29 (1H, s, CH). MS (ESI+) m/e 207 [M+H+].
Intermediate 88




4-Fluoro-2-imidazol-1-yl-benzonitrile. To a solution of imidazole (4.45 g, 65.4 mmol) in tetrahydrofuran (30 mL) and dimethylformamide (10 mL) was added potassium carbonate (9.95 g, 72 mmol) and the mixture was stirred for 30 min at room temp. To this was added 2,4-difluorobenzonitrile (10.0 g, 72 mmol) and the mixture stirred at 90° C. for 3 h then at room temp for 2 days. The mixture was filtered and concentrated and the residue was purified by flash chromatography (SiO2) eluting with 20% to 70% ethyl acetate/hexane to give the title compound as white needles (1.1 g, 9% yield). 1H-NMR (500 MHz, CDCl3) δ ppm: 7.94 (1H, s), 7.84 (1H, dd, J=8.7, 5.6 Hz), 7.37 (1H, t, J=8.7, 5.6 Hz), 7.37 (1H, t, J=1.4 Hz), 7.29 (1H, t, J=1.1 Hz), 7.27-7.21 (2H, m). LCMS [M+H]+ calcd for C10H7N3F: 188.058; found: 188.12.
Intermediate 89




(4-Fluoro-2-(1H-imidazo-1-yl)phenyl)methanamine) hydrochloride. The title compound can be prepared from intermediate 88, 4-fluoro-2-imidazol-1-yl-benzonitrile, according to the method provided for intermediate 79. Yellow solid, 1H-NMR (500 MHz, CD3OD) δ ppm: 9.39 (1H, s), 7.98 (1H, d, J=1.5 Hz), 7.92-7.89 (2H, m), 7.63-7.59 (2H, m), 4.11 (2H, s). LCMS [M+H]+ calcd for C10H11N3F: 192.09; found: 192.15.
Intermediate 90




1-(2-Cyano-5-fluoro-phenyl)-1H-1,2,4-triazole-3-carboxylic acid methyl ester. To a solution of methyl 1H-1,2,4-triazole-3-carboxylate (27 g, 215 mmol) in dimethylformamide (170 mL) was added sodium hydride (5.53 g, 95%, 217 mmol) and the mixture was stirred for 30 min. Added to this was 2,4-difluorobenzylnitrile (30 g, 217 mmol) and the resulting mixture stirred at room temp for 60 h. The mixture was diluted with water and filtered to remove solids. The solution was extracted with ethyl acetate and the organic phase was washed with water (3×'s) and brine, then dried (Na2SO4) and concentrated. The resulting residue was purified by flash chromatography (SiO2) eluting with 30% tetrahydrofuran/20% CH2Cl2/50% hexane to give the title compound as white needles (5.34 g, 10% yield). 1H-NMR (300 MHz, CDCl3) δ ppm: 8.92 (1H, s), 7.85 (1H, dd, J=8.8, 5.5 Hz), 7.67 (1H, dd, J=8.8, 2.6 Hz), 7.34-7.27 (1H, m), 40.3 (3H, s). LCMS [M+H]+ calcd for C11H8N4FO2: 247.06; found: 247.11.
Intermediate 91




Methyl 1-(2-(aminomethyl)-5-fluorophenyl)-1H-1,2,4-triazole-3-carboxylate. The title compound can be prepared from intermediate 90, 1-(2-cyano-5-fluorophenyl)-1H-1,2,4-triazole-3-carboxylic acid methyl ester 1H-NMR (300 MHz, CD3OD) δ ppm: 9.15 (1H, s), 7.80 (1H, dd, J=8.8, 5.9 Hz), 7.71 (1H, dd, J=8.8, 2.6 Hz), 7.46 (1H, td J=8.2, 2.6 Hz), 4.19 (2H, s), 4.03 (3H, s). LCMS [M+H]+ calcd for C11H12N4O2: 251.09; found: 251.17.
Intermediate 92




3-Fluoro-2-(1,1-dioxo-1λ6-[1,2]thiazinan-2-yl)benzonitrile. To a solution of 1,1-dioxo-1λ6-[1,2]thiazin-2-ane (1.90 g, 14.4 mmol) dissolved in tetrahydrofuran (8 mL) and dimethylformamide (2 mL) was added sodium hydride (0.36 g, 95%, 14.4 mmol) and the mixture stirred for 20 min. To this was added 2,3-difluorobenzonitrile (2.0 g, 14.4 mmol) and the mixture stirred at 90° C. for 2 h. The mixture was partitioned between ethyl acetate and water. The organic phase was washed with water and brine then concentrated. The solid residue was triturated with 1:1 ethyl acetate/hexane to give the title compound as a pale brown solid (0.47 g, 13% yield). 1H-NMR (500 MHz, CDCl3) δ ppm: 7.47-7.45 (1H, m), 7.32-7.36 (2H, m), 4.08-4.02 (1H, m), 3.57 (1H, td, J=13.0, 3,7 Hz), 3.40-3.34 (1H, m), 3.32-3.27 (1H, m), 2.44-2.32 (2HF, m), 2.04-1.97 (2H, m), 1.90-1.84 (1H, m). LCMS [M+H]+ calcd for C11H12N2FO2S: 255.28; found: 255.13.
Intermediate 93




3-Fluoro-2-(1,1-dioxo-1λ6-[1,2]thiazinan-2-yl)benzylamine hydrochloride. The title compound can be prepared from intermediate 92, 3-fluoro-2-(1,1-dioxo-1λ6-[1,2]thiazinan-2-yl)benzonitrile according to the procedure provided for intermediate 79. White solid, 1H-NMR (500 MHz, CD3OD) δ ppm: 7.56-7.52 (1H, m), 7.40-7.34 (2H, m), 4.31 (2H, s), 3.98-3.93 (1H, m), 3.68-3.64 (1H, m), 3.42-3.39 (2H, m), 2.42-2.37 (2H, m), 2.03-1.92 (2H, m). LCMS [M+H]+ calcd for C11H16N2O2FS: 259.09; found: 259.18.
Intermediate 94




3-Fluoro-2-1,2,4-triazol-1-yl-benzonitrile. A mixture of 2,3-difluorobenzylnitrile (2.27 g, 16.3 mmol) and triazole sodium salt (1.33 g, 14.8 mmol) in tetrahydrofuran (5 mL) and dimethylformamide (10 mL) was stirred at 85° C. for 4 h. After concentration, the residue was purified by flash chromatography (SiO2) eluting with 25%-50% ethyl acetate/hexane. The isolated product was recrystallized from hot ethyl acetate/hexane to give the title compound as white needles (1.51 g, 54% yield). 1H-NMR (500 MHz, CDCl3) δ ppm: 8.50 (1H, d, J=2.4 Hz), 8.25 (1H, s), 7.69-7.67 (1H, m), 7.60-7.57 (2H, m). LCMS [M+H]+ calcd for C9H6N4F: 189.16; found: 189.14.
Intermediate 95




(3-Fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl)methanamine. The title compound can be prepared from intermediate 94, 3-fluoro-2-1,2,4-triazol-1-yl-benzonitrile. 1H-NMR (500 MHz, CD3OD) δ ppm: 9.61 (1H, d, J=2.9 Hz), 8.79 (1H, s), 7.82-7.74 (1H, m), 7.67-7.57 (2H, m), 4.14-4.13 (2H, m). LCMS [M+H]+ calcd for C9H10N4F: 193.08; found: 193.16.
Intermediate 96




5-Fluoro-2-(1H-1,2,4-triazol-1-yl)benzonitrile. A suspension of 2,5-difluorobenzonitrile (4.5 g, 32.35 mmol) and 1,2,4-triazole sodium salt (3.6 g, 40 mmol) in dimethylformamide (40 mL) was heated at 80° C. for 15 h. The reaction mixture was then cooled, diluted with CH2Cl2 (200 mL), washed with water (3×30 mL) and brine (30 mL), then dried (Na2SO4), filtered and concentrated to give a white solid which was purified by flash column chromatography (SiO2) using 1:1 to 3:1 ethyl acetate/Hexanes to afford the title compound (2.98 g, 49% yield) as a white powder. 1H NMR (500 MHz, CDCl3) δ: 8.70 (1H, s), 8.18 (1H, s), 7.76 (1H, dd, J=9.0, 4.8 Hz), 7.55 (1H, dd, J=7.3, 2.8 Hz), 7.51-7.47 (1H, m). LCMS (M+H) calcd for C9H6FN4: 189.17; found: 189.10.
Intermediate 97




(5-Fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride. A solution of intermediate 96, 5-fluoro-2-(1H-1,2,4-triazol-1-yl)benzonitrile (2.94 g, 15.59 mmol) in ethanol (100 mL) and 1N HCl (50 mL) was degassed by bubbling N2. Then, 10% Pd/C was added, the flask evacuated and vented to H2 three times and left on a Parr shaker under a H2 atmosphere (40 psi). After 6 h, the reaction mixture was filtered, concentrated and the aqueous solution lyophilized to afford the title compound (4.07 g, 98%) as a white powder. LCMS (M+H) calcd for C9H10FN4: 193.09; found: 193.15.
Intermediate 98




2-(1H-1,2,4-Triazol-1-yl)benzonitrile. A suspension of 2-fluorobenzylnitrile (3.0 g, 25 mmol) and 1,2,4-triazole sodium salt (2.4 g, 27 mmol) were stirred in tetrahydrofuran (7 mL) and dimethylformamide (14 mL) at 95° C. for 18 h. After cooling and concentrating, the product was crystallized from hot CH2Cl2/hexane (1:1) to give the title compound as a white solid (4.25 g, 100% yield). 1H-NMR (300 MHz, CDCl3) δ ppm: 8.74 (1H, s), 8.16 (1H, s), 7.82 (1H, dd, J=4.9, 1.3 Hz), 7.77-7.25 (2H, m), 7.57-7.51 (1H, m). LCMS [M+H]+ calcd for C9H7N4: 171.06; found: 171.12.
Intermediate 99




(2-(1H-1,2,4-Triazol-1-yl)phenyl)methanamine hydrochloride. Intermediate 98, 2-(1H-1,2,4-triazol-1-yl)benzonitrile (4.25 g, 25 mmol) was dissolved in ethanol (50 mL) and 1N HCl (25 mL). 10% Pd—C (1 g) was added and the mixture shaken under H2 for 2 h at 50 psi. After filtration through Celite and concentration, the residue was triturated with diethyl ether and the title compound was collected as a white solid. (3.94 g, 75% yield). 1H-NMR (300 MHz, CD3OD) δ ppm: 9.01 (1H, s), 8.32 (1H, s), 7.78-7.64 (4H, m), 4.15 (2H, s). LCMS [M+H]+ calcd for C9H11N4: 175.09; found: 175.17.
Intermediate 100




2-(1,1-Dioxo-1λ6-[1,2]thiazinan-2-yl)benzonitrile. Added to a solution of 1,1-dioxo[1,2]thiazinane (3.37 g, 25 mmol) in dimethylformamide (35 mL) was sodium hydride (0.675 g, 25 mmol, 95%) and the mixture stirred at room temperature for 15 min. 2-Fluorobenzonitrile (3.37 mL, 31.3 mmol) was added and the mixture stirred at 80° C. for 18 h. The mixture was cooled, diluted with water and extracted with ethyl acetate. The organic phase was washed with water and brine, then dried (Na2SO4) and concentrated. The residue was purified by flash chromatography (SiO2) eluting with 10%-100% ethyl acetate/hexane. The isolated solid was recrystallized from hot ethyl acetate/hexane (2:1) to give the title compound as white crystals (4.15 g, 70% yield). 1H-NMR (300 MHz, CDCl3) δ ppm: 7.70 (1H, dd, J=7.7, 1.1 Hz), 7.64-7.53 (2H, m), 7.41 (1H, td, J=7.3, 1.6 Hz), 3.72 (2H, t, J=5.5 Hz), 3.32 (2H, t, J=6.0 Hz), 2.40-2.32 (2H, m), 2.05-1.97 (2H, m). LCMS [M+H]+ calcd for C11H12N2O2S: 237.06; found: 237.10.
Intermediate 101




2-(1,1-Dioxo-1λ6-[1,2]thiazinan-2-yl)benzylamine hydrochloride. Intermediate 100, 2-(1,1-dioxo-1λ6-[1,2]thiazinan-2-yl)benzonitrile, (2.63 g, 11.14 mmol) was dissolved in ethanol (150 mL) and 1N HCl (13 mL). Added to this was 10% Pd—C (0.5 g) and the mixture shaken under H2 at 55 psi for 24 h. Filtration through Celite followed by concentration gave the title compound as a white solid (2.93 g, 95% yield). 1H-NMR (300 MHz, CD3OD) δ ppm: 7.61-7.47 (4H, m), 4.30 (2H, q, J=13.7 Hz), 3.96-3.87 (1H, m), 3.49-3.36 (3H, m), 2.40-2.31 (2H, m), 2.05-1.96 (2H, m). LCMS [M+H]+ calcd for C11H17N2SO2: 241.10; found: 241.10.
Intermediate 102




(3,5-Difluoropyridin-2-yl)methanamine hydrochloride. A mixture of 3,5-difluoropicolinonitrile (1.4 g, 10 mmol), conc. HCl (12 ml) and 10% Pd—C (200 mg) in 1:1 ethanol/tetrahydrofuran was shaken under a hydrogen atmosphere (50 psi) for 5 h. The reaction mixture was filtered and the ethanol removed in vacuo. The remaining solution was lyophilized to afford an off-white solid (2.16 g, 100% yield). LCMS (M+H) calcd for C6H7F2N2: 145.06; found: 145.12.
Intermediate 103




(5-Chloropyridin-2-yl)methanamine. A solution of 5-chloropicolinonitrile (3.8 g, 27.43 mmol), conc. HCl (3 mL) and 10% Pd—C (1.0 g) in ethanol (100 mL) was shaken under a hydrogen atmosphere (40 psi) for 2 h. The reaction mixture was filtered, concentrated and the resulting residue taken up in satd NaHCO3 (50 mL) and extracted with CH2Cl2 (4×25 mL). The combined CH2Cl2 layers were dried (Na2SO4), filtered and concentrated to give the title compound as a yellow oil (2.0 g, 51% yield). LCMS (M+H) calcd for C6H8ClN2: 143.04; found: 143.07. 1HNMR (500 MHz, CDCl3) δ ppm: 8.56-8.51 (1H, br d), 7.66-7.60 (1H, m), 7.28-7.14 (1H, m), 3.97 (2H, s), 1.72 (2H, s).
Intermediate 104




2-(Bromomethyl)-5-fluorobenzonitrile. N2 was passed through a mixture of 5-fluoro-2-methylbenzonitrile (28.51 g, 211 mmol), NBS (41.31 g, 232 mmol) and AIBN (2.5 g, 15 mmol) in CCl4 (845 mL) for 10 min after which the reaction was heated at reflux for 8 h. After standing at room temperature overnight, the reaction mixture was filtered and the filter cake washed with CCl4 (500 mL). The combined filtrate was evaporated to give a yellow oil. Flash chromatography (SiO2) using 5-25% ethyl acetate/Hexanes as eluent afforded the title compound (29.74 g, 66% yield) as a pale yellow oil. 1H NMR (500 MHz, CDCl3) δ: 7.55 (1H, dd, J=8.6, 5.2 Hz), 7.37 (1H, dd, J=7.9, 2.8 Hz), 7.32-7.28 (1H, m), 4.61 (2H, s).
Intermediate 105




2-((1,3-Dioxoisoindolin-2-yl)methyl)-5-fluorobenzonitrile. To a stirred solution of intermediate 104, 2-(bromomethyl)-5-fluorobenzonitrile (29.72 g, 139 mmol) and phthalimide (32.69 g, 222 mmol) in dimethylformamide (300 mL) was added Cs2CO3 (67.87 g, 208 mmol). After stirring vigorously for 1 h, the reaction mixture was poured into water (1.2 L). The precipitated product was filtered, washed with water (600 mL) and methanol (150 mL) to give a white solid. The solid was taken up into 1 L of water/methanol (2:1) to which was added K2CO3 (12 g) and the mixture stirred at 40° C. After 30 min., the mixture was cooled and filtered. The filter cake was washed with water (500 mL), and dried under vacuum to afford the title compound (38.91 g, 94% yield) as a white powder. 1H NMR (500 MHz, CDCl3) δ: 7.89 (2H, dd, J=5.5, 3.1 HZ), 7.76 (5.5, 3.1 Hz), 7.41 (1H, dd, J=8.6, 5.2 Hz), 7.38 (1H, dd, J=7.9, 2.8 Hz), 7.24 (1H, td, J=8.2, 2.8 Hz), 5.06 (2H, s). LCMS (M+H) calcd for C16H10FN2O2: 281.07; found: 281.15.
Intermediate 106




tert-Butyl 2-cyano-4-fluorobenzylcarbamate. A suspension of intermediate 105, 2-((1,3-dioxoisoindolin-2-yl)methyl)-5-fluorobenzonitrile, (5.6 g, 20 mmol) in dimethylformamide (20 mL) was warmed until it was dissolved. To this was added tetrahydrofuran (100 mL) and the mixture placed in a pre-heated (70° C.) oil bath. Hydrazine monohydrate was added to this and the reaction stirred for 8 h. The resulting white slurry was left at ambient temperature overnight. To this slurry was added di-tert-butyldicarbonate (6.55 g, 30 mmol) and the mixture stirred for 6 h at room temperature. The reaction mixture was diluted with ether (100 mL), filtered and the filtrate treated with activated carbon at 40° C. After filtration and concentration the crude product was purified by flash chromatography, using 20-30% ethyl acetate/Hexanes as eluent, to provide the title compound (2.88 g, 58% yield) as a light yellow powder. 1H NMR (500 MHz, CDCl3) δ: 9.46 (1H, br s), 7.61 (1H, dd, J=7.9, 2.1 Hz), 7.34 (1H, dd, J=8.2, 4.6 Hz), 7.22 (1H, td, J=8.6, 2.4 Hz), 4.71 (2H, s), 1.59 (9H, s). LCMS (M+H) calcd for C13H16FN2O2: 251.12; found: 251.22.
Intermediate 107




2-(Aminomethyl)-5-fluorobenzonitrile trifluoroacetic acid salt. A round-bottom flask was charged with intermediate 106, tert-butyl 2-cyano-4-fluorobenzylcarbamate, (1.9 g, 7.591 mmol) then treated with trifluoroacetic acid (20 ml) at room temperature. After 1 h, the reaction mixture was concentrated to give a yellow oil which was dissolved in CHCl3 and re-concentrated to afford the title compound (2.01 g, 100% yield) as a pale yellow solid. LCMS (M+H) calcd for C8H8FN2: 151.07; found: 151.08.
Intermediate 108




(2,5-Dibromo-4-fluorophenyl)methanamine. A solution of 2,5-dibromo-4-fluorobenzyl bromide (0.350 g, 1 mmol) in 7M NH3/MeOH was heated in a sealed tube at 100° C. for 2 h. The reaction mixture was cooled and concentrated to give a white solid which was dissolved in CH2Cl2 and treated with Et3N (1 mL) then concentrated. The resulting residue was triturated with ethyl acetate (25 mL), filtered and concentrated to give the title compound (0.291 g) as a pale yellow oil. HRMS (M+H) calcd for C7H7Br2FN: 283.94; found: 283.93.
Intermediate 109




4-Fluoro-2-methylsulfanyl-benzylamine. Under N2, 4-fluoro-2-(methylthio)benzonitrile (1.67 g, 0.1 mol) was dissolved in 20 mL tetrahydrofuran and treated with 10 mL 2M BH3.Me2S. This was heated at 60° C. for 2 hrs. Heating was discontinued and 5 mL MeOH was cautiously added, followed by the cautious addition of 4 mL 6N HCl. Additional H2O (20 mL) was added followed by ethyl acetate. The layers were separated. The aqueous layer was made basic with 1N NaOH and extracted with CH2Cl2. The extracts were dried (MgSO4), filtered, concentrated and dried in vacuo to give 1.3 g of the title compound as a solid (Yield 76%). 1H NMR (500 MHz, CDCl3) δ ppm: 7.20-7.31 (1H, m) 6.90 (1H, dd, J=2.4 Hz) 6.75-6.86 (1H, m) 3.86 (2H, s) 2.47 (3H, s); LC/MS m/z 172.
Intermediate 110




2-(Aminomethyl)-5-fluorobenzenamine hydrochloride. 2-Amino-4-fluorobenzonitrile (Fritz Hunziker et al. Eur. J. Med. Chem. 1981, 16, 391) (0.300 g, 1.68 mmol), was dissolved in acetic anhydride (5 mL) and the solution was stirred at 23° C. for 18 h. An additional portion of acetic anhydride (3 mL) was added to dissolve the N-(2-cyano-5-fluorophenyl)acetamide. Then palladium (10% on charcoal) (25 mg) was added and the mixture was agitated under H2 (34 psi) for 72 h. The Pd—C was removed by filtration on Celite and the filtrate concentrated in vacuo to afford a bis-acetamide: LCMS (M+H)+ m/z 225. This was heated at reflux with HCl (6N, 10 mL) for 30 min. The acid was removed under reduced pressure to give a solid which was crystallized from MeOH-ether to afford the title compound (0.120 g, 51% yield). 1H NMR (400 MHz, MeOD) δ ppm: 7.51 (1H, m), 6.96 (2H, m), 4.20 (2H, s).
Intermediate 111




4-Fluoro-2-(2-oxopyrrolidin-1-yl)benzonitrile. A 48 mL pressure vessel containing 2-bromo-4-fluorobenzonitrile (1.00 g, 5.00 mmol), 2-pyrrolidinone (0.46 mL, 6.00 mmol), Cs2CO3 (2.28 g, 7.0 mmol) and 9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene (xantphos) (0.231 g, 0.40 mmol) in dioxane (6 mL) was degassed with argon for 15 min. Pd2 dba3 was introduced and the reaction mixture heated at 105° C. for 48 h. The mixture was cooled, diluted with ethyl acetate or dioxane, and then filtered through Celite. The resulting mixture was concentrated in vacuo and subjected to column chromatography on silica gel with hexanes:ethyl acetate (3:7) gradient as the eluent to afford the title compound as a white solid (0.887 g, 87% yield): 1H NMR (400 MHz, CDCl3) δ ppm: 7.69 (1H, dd, J=5.8, 8.6 Hz), 7.22 (1H, dd, J=2.5, 9.6 Hz), 7.07 (1H, ddd, J=2.5, 7.6, 8.6 Hz), 3.96 (2H, t, J=7.0 Hz), 2.62 (2H, t, J=8.1 Hz), 2.30-2.22 (2H, m); LCMS (+ESI, M+H+) m/z 205.
Intermediate 112




4-Fluoro-2-(2-oxopiperidin-1-yl)benzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111 1H NMR (400 MHz, CDCl3) δ ppm: 7.71 (1H, dd, J=5.7, 8.7 Hz), 7.14-7.06 (1H, m), 7.08 (1H, dd, J=2.4, 9.0 Hz), 3.65 (2H, t, J=5.7 Hz), 2.60 (2H, t, J=6.3 Hz), 2.05-1.95 (4H, m); LCMS (+ESI, M+H+) m/z 219.
Intermediate 113




4-Fluoro-2-(2-oxoazepan-1-yl)benzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111. 1H NMR (400 MHz, CDCl3) δ ppm: 7.68 (1H, dd, J=5.8, 8.6 Hz), 7.08 (1H, ddd, J=2.5, 7.6, 8.6 Hz), 7.01 (1H, dd, J=2.5, 9.0 Hz), 3.77-3.76 (2H, m), 2.75-2.72 (2H, m), 1.91-1.86 (6H, m); LCMS (+ESI, M+H+) m/z 233.
Intermediate 114




N-(2-Cyano-5-fluorophenyl)-N-methylacetamide. The title compound can be prepared according to the procedure provided for intermediate 111 1H NMR (400 MHz, CDCl3) δ ppm: 7.79-7.75 (1H, m), 7.32-7.19 (1H, m), 7.10-7.07 (1H, m), 3.42 (0.6H, brs), 3.30 (2.4H, s), 2.32 (0.6H, brs), 1.91 (2.4H, s); LCMS (+ESI, M+H+) m/z 193; HPLC: 94% (220 nm).
Intermediate 115




2-(2-Oxoazetidin-1-yl)benzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111. 1H NMR (400 MHz, DMSO-d6) δ ppm: 8.02 (1H, d, J=8.4 Hz), 7.76 (1H, dd, J=1.5, 7.8 Hz), 7.69-7.65 (1H, m), 7.23 (1H, s), 4.04 (2H, t, J=4.8 Hz), 3.16 (2H, t, J=4.8 Hz). LCMS (+ESI, M+H+) m/z 173.
Intermediate 116




2-(2-Oxooxazolidin-3-yl)benzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111. 1H NMR (400 MHz, CDCl3) δ ppm: 7.71 (1H, dd, J=1.5, 7.6 Hz), 7.68-7.63 (1H, m), 7.58 (1H, d, J=7.6 Hz), 7.38 (1H, dt, J=1.3, 7.6 Hz), 4.57 (2H, t, J=7.8 Hz), 4.21 (2H, t, J=7.8 Hz); LCMS (+ESI, M+H+) m/z 189.
Intermediate 117




4-Fluoro-2-(2-oxooxazolidin-3-yl)benzonitrile. A 48 mL pressure vessel containing 2-bromo-4-fluorobenzonitrile (1.00 g, 5.00 mmol), 2-oxazolidone (0.390 g, 4.50 mmol), K2CO3 (0.970 g, 7.0 mmol) and xantphos (0.231 g, 0.40 mmol) in dioxane (10 mL) was degassed with argon for 15 min. Pd2 dba3 (0.140 g, 0.15 mmol) was introduced and then the reaction mixture was heated at 70° C. for 18 h. The mixture was cooled, diluted with dioxane, and then filtered through Celite. The resulting mixture was concentrated in vacuo and subjected to column chromatography on silica gel with hexanes:ethyl acetate (1:1) to (3:7) gradient as the eluent to afford the title compound as a white solid (0.460 g, 50% yield): 1H NMR (400 MHz, CDCl3) δ ppm: 7.73 (1H, dd, J=5.8, 8.6 Hz), 7.43 (1H, dd, J=2.5, 9.6 Hz), 7.11 (1H, ddd, J=2.5, 7.5, 8.7 Hz), 4.60 (2H, t, J=7.1 Hz), 4.29 (2H, t, J=7.1 HJz); LCMS (+ESI, M+H+) m/z 207.
Intermediate 118




3-(2-(Aminomethyl)-5-fluorophenyl)oxazolidin-2-one hydrochloride. 1H NMR (400 MHz, MeOD) δ ppm: 7.73 (1H, dd, J=6.0, 8.6 Hz), 7.43 (1H, dd, J=2.5, 9.5 Hz), 7.11 (1H, ddd, J=2.5, 7.5, 8.6 Hz), 4.64 (2H, t, J=7.7 Hz), 4.17 (2H, t, J=7.7 Hz), 4.14 (2H, s); LCMS (+ESI, M+H+) m/z 211.
Intermediate 119




4-Fluoro-2-(2-oxoazetidin-1-yl)benzonitrile. The title compound can be prepared according to the procedure provided for intermediate 117 1H NMR (400 MHz, CDCl3) δ ppm: 8.06 (1H, dd, J=10.7, 2.6 Hz), 7.58 (1H, dd, J=8.6, 6.3 Hz), 7.87 (1H, td, J=8.6, 2.5 Hz), 4.25 (2H, t, J=5.0 Hz), 3.26 (2H, t, J=5.0 Hz); LCMS (+ESI, M+H+) m/z 191.
Intermediate 120




1-(2-(Aminomethyl)-5-fluorophenyl)azetidin-2-one hydrochloride. 1H NMR (400 MHz, DMSO/D2O) δ ppm: 7.54 (1H, dd, (t), J=8.6 Hz), 7.25 (1H, dd, J=10.8, 2.5 Hz), 7.17 (1H, td, J=8.6, 2.5 Hz), 4.12 (2H, s), 3.79 (2H, t, J=4.6 Hz), 3.09 (2H, t, J=4.6 Hz); LCMS (+ESI, M+H+) m/z 195.
Intermediate 121




(R)-2-(2-((tert-Butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111. 1H NMR (400 MHz, CDCl3) δ ppm: 7.68 (1H, dd, J=5.8, 8.8 Hz), 7.19 (1H, dd, J=2.5, 9.1 Hz), 7.11-7.07 (1H, m), 4.46-4.42 (1H, m), 3.55 (2H, d, J=3.3 Hz), 2.72-2.52 (2H, m), 2.43-2.33 (1H, m), 2.09-2.01 (1H, m), 0.81 (9H, s), −0.04 (3H, s), −0.07 (3H, s); LCMS (+ESI, M+H+) m/z 349.
Intermediate 122




(S)-2-(2-((tert-Butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111. 1H NMR (400 MHz, CDCl3) δ ppm: 7.68 (1H, dd, J=5.8, 8.6 Hz), 7.19 (1H, dd, J=2.5, 9.4 Hz), 7.11-7.07 (1H, m), 4.46-4.43 (1H, m), 3.55 (2H, d, J=3.3 Hz), 2.72-2.52 (2H, m), 2.43-2.33 (1H, m), 2.09-2.01 (1H, m), 0.81 (9H, s), −0.04 (3H, s), −0.07 (3H, s); LCMS (+ESI, M+H+) m/z 349.
Intermediate 123




4-Fluoro-2-(thiazol-2-ylamino)benzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111. 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.21 (1H, s), 8.39-8.35 (1H, m), 7.97 (1H, d, J=5.0 Hz), 7.23-7.13 (3H, m); LCMS (+ESI, M+H+) m/z 220.
Intermediate 124




4-Fluoro-2-(5-methyl-1,3,4-thiadiazol-2-ylamino)benzonitrile. The title compound can be prepared according to the procedure provided for intermediate 111. 1H NMR (400 MHz, DMSO-d6) δ ppm: 8.30 (1H, dd, J=6.5, 8.8 Hz), 7.96 (1H, s), 7.26-7.19 (2H, m), 2.64 (3H, s); LCMS (+ESI, M+H+) m/z 235.
Intermediate 125




1-(2-(Aminomethyl)-5-fluorophenyl)piperidin-2-one hydrochloride salt. To a stirred solution of intermediate 112, 4-fluoro-2-(2-oxopiperidin-1-yl)benzonitrile (150 mg, 0.69 mmol) in H2O (10 mL) was added ethanol (10 mL) 10% palladium on charcoal (50 mg) and 1N HCl (2.1 mL, 20.6 mmol). The reaction was shaken in a Parr system under H2 (40 psi) for 1 h. Then the Pd/C catalyst was removed by filtration on Celite and the filtrate was concentrated in vacuo to yield a solid. Toluene (2×50 mL) was added to the solid and the solution was evaporated in vacuo. LCMS (M+H)+ m/z 170.
Intermediate 126




1-Bromo-4-fluoro-2-methoxybenzene. To a mixture of 2-bromo-5-fluorophenol (10 g, 50.8 mmol) and iodomethane (11.2 g, 78.7 mmol) in dimethylformamide (100 mL) was added potassium carbonate (10.9 g, 79 mmol) and the mixture stirred at room temperature for 3 hrs. The mixture was diluted with water (100 mL) and extracted with ether (50 mL×3). The combined extracts were washed with brine, dried over anhydrous magnesium sulfate, and concentrated in vacuo to obtain 11.3 g of 1-bromo-4-fluoro-2-methoxybenzene as an amber colored oil.
Intermediate 127




4-Fluoro-2-methoxybenzonitrile. To a solution of intermediate 126, 1-bromo-4-fluoro-2-methoxybenzene (9.0 g) in N-methylpyrrolidone (100 mL, Sure Seal; Aldrich) was added CuCN (6.6 g, 73.7 mmol, 1.8 eq.; Aldrich), and the mixture stirred at 180° C. under anhydrous nitrogen for 5.5 hrs. After cooling, 14% aqueous NH4OH (330 mL) was added and stirring continued for 45 min at room temperature. The mixture was extracted with ether (100 mL×3), and the combined extracts washed sequentially with dilute aqueous NH4OH, dilute HCl and brine, then dried (MgSO4), and concentrated to provide the title compound (5.2 g, Yield 85% in 2 steps) as a white solid: 1H NMR (CDCl3, 500 MHz) δ ppm: 3.91 (3H, s, OMe), 6.69 (1H, dd, J=2.3 Hz, J=10.5 Hz, Ar—H), 6.72 (1H, dt, J=2.5 Hz, J=J=8.0 Hz, Ar—H), 7.55 (1H, dd; J=6.5 Hz, J=8.5 Hz, Ar—H); 13C NMR (CDCl3, 125.8 Hz) δ ppm: 56.49, 98.16, 100.06, 100.27, 108.31, 108.50, 115.83 135.37, 135.46, 163.25, 163.34 165.47, 167.50. An analytical sample was obtained by trituration with ether: Anal. calcd for C8H6FNO: C, 63.57; H, 4.00; N, 9.26; found: C, 63.36; H, 3.91; N, 9.16.
Intermediate 128




4-Fluoro-2-methoxybenzylamine hydrochloride. To a mixture of intermediate 127, 4-fluoro-2-methoxybenzonitrile, (800 mg, 5.3 mmol) and conc.HCl (0.53 mL, 6.36 mmol, 1.2 eq.) in ethanol (20 mL) was added 10% Pd—C (100 mg; Aldrich), and the mixture hydrogenated at 1 atm hydrogen for 15 hrs at room temperature. To this mixture was added an additional amount of conc.HCl (1 mL) and 10% Pd—C (200 mg) and the reaction allowed to continue for another 40 hrs. The mixture was filtered through Celite and the filtrate concentrated in vacuo to dryness. The residue was triturated with ether to provide the title compound (895 mg, Yield 88%) as a white powder: 1H NMR (CDCl3, 500 MHz) δ ppm: 3.84 (3H, s, OMe), 3.91 (2H, d, J=5.5 Hz, N—CH2), 6.81 (1H, dt, J=2.5 Hz, J=J=8.5 Hz, Ar—H), 6.99 (1H, dd, J=2.5 Hz, J=11.3 Hz, Ar—H), 7.47 (1H, dd, J=7 Hz, J=8.5 Hz, Ar—H); 13C NMR (CDCl3, 125.8 Hz) δ ppm: 36.76, 56.03, 99.30, 99.51 106.28, 106.45, 117.93, 117.95, 131.60, 131.69, 158.56, 158.64, 162.28, 164.22. HRMS (ESI) calcd for C8H11FNO (M+H) 156.0825, found 156.0830.
Intermediate 129




4-Fluoro-2-hydroxybenzonitrile. A mixture of intermediate 127, 4-fluoro-2-methoxybenzonitrile, (4.53 g, 30 mmol;) and AlCl3 (5.0 g, 37.6 mmol; Aldrich) in anhydrous toluene (30 mL) was stirred at approximately 130° C. for 18 hrs. After cooling, ice water ( ̃50 mL) was added and the resulting mixture extracted with ether (20 mL×2). The combined extracts were washed sequentially with water and brine, then dried (MgSO4), and concentrated in vacuo to provide the title compound (3.90 g, 28.5 mmol, Yield 95%) as a white solid: 1H NMR (DMSO-d6, 300 MHz) δ ppm: 6.74-6.84 (2H, m, Ar—Hs), 7.71 (1H, dd, J=7 Hz, J=8.5 Hz, Ar—H), 11.64 (1H, s, OH); 13C NMR (DMSO-d6, 75.5 Hz) δ ppm: 95.13 102.45, 102.78, 106.53, 106.83 115.53, 134.68, 134.84, 161.41, 161.58, 163.00, 166.35.HRMS (ESI−) calcd for C7H3NOF (M−H) 136.0199, found 136.0199.
Intermediate 130




4-Fluoro-2-(2-morpholino-2-oxoethoxy)benzonitrile. To a solution of intermediate 129, 4-fluoro-2-hydroxybenzonitrile, (685 mg, 5 mmol) in dimethylformamide (8 mL, Sure Seal; Aldrich) was added NaH (200 mg, 5 mmol; 60% oil dispersion; Aldrich), and the mixture stirred for 5 min under an anhydrous nitrogen atmosphere. To this was added 4-(2-chloroacetyl)morpholine (900 mg, 5.5 mmol, 1.1 eq.; Avocado Organics), and stirring continued at room temperature for 21 hrs. The reaction was quenched by careful addition of water (30 mL). The resulting mixture was extracted with CH2Cl2 (25 mL×2). The combined extracts were washed with brine, dried (MgSO4) and concentrated. The residue was triturated to obtain 1.10 g (4.17 mmol, Yield 83%) of the title compound as a white solid: 1H NMR (CDCl3, 500 MHz) δ ppm: 3.63 (2H, t, J=4 Hz, NCH2), 3.67 (1H, m, OCH), 3.72 (1H, m, OCH), 4.86 (2H, s, OCH2), 6.80-6.86 (2H, m, Ar—Hs), 7.61 (1H, dd, J=8.5 Hz, 6.1 Hz, Ar—H); 13C NMR (CDCl3, 125.77 Hz) δ ppm: 42.63, 46.04, 66.80, 68.33, 98.45, 98.47, 101.57, 101.79, 109.56, 109.74, 115.42, 135.48, 135.57, 161.26, 161.35, 114.79, 165.23, 167.28. HRMS calcd for C13H14N2O3F (M+H) 265.0988, found 265.0998.
Intermediate 131




2-(2-(Aminomethyl)-5-fluorophenoxy)-1-morpholinoethanone hydrochloride. A solution of intermediate 130, 4-fluoro-2-(2-morpholino-2-oxoethoxy)benzonitrile, (500 mg, 1.89 mmol) in warm ethanol (30 mL) and ethyl acetate (30 mL) was mixed with conc.HCl (0.32 mL, 3.78 mmol, 2 eq.). To this was added 10% Pd—C (100 mg; Aldrich), and the mixture was hydrogenated at 1 atm of hydrogen for 20 hrs at room temperature. To this mixture was added an additional amount of 10% Pd—C (50 mg) and stirring continued for another 7 hrs. The mixture was filtered through Celite and the filtrate concentrated in vacuo to dryness. The residue was triturated with ethyl acetate, then with ethanol to obtain the title compound (168 mg, Yield 29%) as an off-white powder: 1H NMR (CD3OD, 500 MHz) δ ppm: 3.55 (2H, t, J=5 Hz, NCH2), 3.62 (2H, t, J=5 Hz, NCH2), 3.70 (2H, t, J=5 Hz, OCH2), 3.75 (2H, t, J=5 Hz, OCH2), 4.17 (2H, s, NCH2), 5.17 (2H, s, OCH2), 6.82 (1H, dt, J=2.5, 8.5 Hz, Ar—H), 7.05 (1H, dd, J=2.5, 10.5 Hz, Ar—H), 7.43 (1H, dd, J=6.5, 8.5 Hz, Ar—H); 13C NMR (CD3OD, 125.77 Hz) δ ppm: 39.40, 42.49, 44.97, 66.11, 66.46, 66.59, 101.38, 101.59, 108.40, 108.57, 118.40, 132.53, 132.62, 158.43, 158.52, 63.87, 165.83, 168.27. HRMS (ESI) calcd for C13H18N2O3F (M+H) 269.1301, found 269.1301.
Intermediate 132




Dimethyl-carbamic acid 2-cyano-5-fluoro-phenyl ester. Under N2, a stirred solution of intermediate 129, 4-fluoro-2-hydroxybenzonitrile (685 mg, 5.00 mmol), dimethylcarbamoyl chloride, and triethylamine (606 mg, 6 mmol) in dichloroethane (10 mL) was heated at reflux for 20 hrs. The cooled mixture was diluted with dichloroethane (10 mL) washed with water, and brine. The organic layer was separated, dried (Na2SO4), concentrated, and the residue purified by column chromatography (SiO2, 5% ethyl acetate-CH2Cl2) to provide 700 mg (Yield 67%) of the title compound as a white crystalline solid: 1H NMR (CDCl3, 500 MHz) δ ppm: 3.03 (3H, s, NMe), 3.15 (3H, s, NMe), 6.99 (1H, dt, J=2.5 Hz, 8.5 Hz, Ar—H), 7.23 (1H, dd, J=2.5 Hz, 9.5 Hz, Ar—H), 7.61 (1H, dd, J=9 Hz, 6 Hz, Ar—H); 13C NMR (CDCl3, 125.77 Hz) δ ppm: 36.76, 37.06, 102.84, 102.86, 111.59, 111.79, 113.24, 113.42, 114.99, 134.36, 134.45, 152.54, 155.06, 155.16, 164.26, 166.31. HRMS (ESI) calcd for C10H10N2O2F (M+H) 209.0726, found 209.0722.
Intermediate 133




Dimethyl-carbamic acid 2-aminomethyl-5-fluoro-phenyl ester hydrochloride. To a solution of intermediate 132, dimethyl-carbamic acid 2-cyano-5-fluoro-phenyl ester, (340 mg, 1.63 mmol) in ethyl acetate (20 mL) and ethanol (20 mL), was added conc.HCl (0.4 mL) and 10% Pd—C (100 mg) and the mixture hydrogenated in a Parr Shaker at 55 psi of hydrogen for 20 hrs. The reaction mixture was filtered through Celite, and the filtrate concentrated in vacuo to give an oil which was partitioned between ethyl acetate (10 mL) and water (10 mL). After separation, the aqueous phase was washed with additional ethyl acetate (5 mL). The combined extracts were concentrated in vacuo to dryness. The residual oil was triturated with ether to provide 145 mg (Yield 38%) of the title compound, as a tan powder: 1H NMR (CD3OD, 500 MHz) δ ppm: 3.06 (3H, s, NMe), 3.21 (3H, s, NMe), 4.11 (2H, s, NCH2), 7.13 (2H, m, Ar—Hs), 7.60 (1H, m, Ar—H); 13C NMR (CD3OD, 125.77 Hz) δ ppm: 36.03, 36.25 37.58, 110.79, 110.99, 113.26, 113.43, 122.32, 132.18, 132.25, 151.55, 154.72, 162.69, 164.67. HRMS (ESI) calcd for C10H13N2O2F (M+H) 213.1039, found 213.1039.
Intermediate 134




2-(Benzyloxy)-4-fluorobenzonitrile. Benzyl alcohol (13 mL, 125 mmol) was slowly added to a stirred suspension of NaH (95%, 2.86 g, 113 mmol) in toluene (200 mL) at room temperature. After 30 min, 2,4-difluorobenzonitrile (15.3 g, 1100 mmol; Aldrich) was added all at once and stirring continued overnight (18 h). After this, the reaction mixture was washed with water (2×25 mL) and brine (25 ml). The organic layer was dried (Na2SO4), filtered and concentrated to give a white slurry which was triturated with hexanes and filtered to afford the title compound as a white solid (20.34 g, 81% yield). 1H NMR (500 MHz, CDCl3): 7.59-7.55 (1H, m), 7.45-7.34 (5H, m), 6.75-6.71 (2H, m), 5.19 (2H, s); 13C NMR (125.76 MHz, DMSO-d6) δ ppm: 71.16, 98.75, 101.54, 101.75, 108.66, 108.84, 115.83, 127.16, 128.58, 128.94, 135.03, 135.44, 135.54, 162.22, 162.31, 165.26, 167.29. LCMS calcd for C14H11FNO: 228.2; found: 228.0.
Intermediate 135




2-Hydroxy-4-fluoro-benzylamine hydrochloride. A solution of intermediate 134, 2-(benzyloxy)-4-fluorobenzonitrile, (9.03 g, 39.7 mmol) in ethanol (100 mL) and ethyl acetate (100 mL) was stirred with 10% palladium on carbon (1.67 g,) and concentrated hydrochloric acid (12 mL, 144 mmol) under a hydrogen atmosphere (60 psi) for four days. The catalyst was removed by filtration through Celite, and the filtrate was concentrated. The crude product was triturated with ether and the resulting solid collected by filtration to give the title compound (5.24 g, 74% yield) as a pale orange solid. 1H NMR (500 MHz, DMSO-D6) δ ppm: 10.81 (1H, s), 8.18 (3H, s), 7.36 (1H, t, J=7.3 Hz), 6.79 (1H, dd, J=10.8, 2.6 Hz), 6.66 (1H, dt, J=8.5, 2.3 Hz), 3.90 (2H, d, J=5.2 Hz).
Intermediate 136




(2,2-Diethoxyethyl)(o-tolyl)sulane. In ethanol (50 mL) was dissolved sodium metal (1.6 g, 66 mmol) at 23° C. 2-Methylbenzenethiol (8.1 mL, 68 mmol) was slowly added to this solution, followed by bromoacetaldehyde diethylacetal (9.50 mL, 63 mmol). The reaction mixture was stirred at reflux for 18 h. The solvent was then evaporated in vacuo and the residue was washed with H2O (100 mL) and extracted with ether (100 mL). The organic solution was dried (MgSO4), concentrated in vacuo and purified by distillation to afford the title compound (13.48 g, 82% yield): 1H NMR (400 MHz, CDCl3) δ ppm: 7.33 (1H, d, J=7.9 Hz), 7.16-7.08 (3H, m), 4.65 (1H, t, J=5.6 Hz), 3.66 (2H, q, J=7.0 Hz), 3.55 (2H, q, J=7.0 Hz), 3.09 (2H, d, J=5.6 Hz), 2.38 (3H, s), 1.20 (6H, t, J=7.0 Hz). LCMS (M+H)+ m/z 241 (t=2.65 min.).
Intermediate 137




7-Methylbenzo[b]thiophene. To a solution of intermediate 136, (2,2-diethoxyethyl)(o-tolyl)sulfane (0.58 g, 2.41 mmol) in chlorobenzene (20 mL) was added polyphosphoric acid. The reaction mixture was stirred at reflux for 18 h. Water (100 mL) was then added and the organic material was extracted with CH2Cl2 (2×50 mL). The organic solution was dried (MgSO4) and concentrated in vacuo to afford 335 mg (94% yield) of the title compound: 1H NMR (400 MHz, CDCl3) δ: 7.68 (1H, d, J=7.8 Hz), 7.43 (1H, d, J=5.4 Hz), 7.36 (1H, d, J=5.4 Hz), 7.30 (1H, dd, J=7.8, 7.1 Hz), 7.14 (1H, d, J=7.1 Hz), 2.58 (3H, s); LCMS (M+H)+ m/z 148.
Intermediate 138




7-(Bromomethyl)benzo[b]thiophene. To a solution of intermediate 137, 7-methylbenzo[b]thiophene (1.0 g, 6.5 mmol) in CCl4 (20 mL) was added benzoyl peroxide (1.1 g, 4.54 mmol) followed by portionwise addition of NBS (1.15 g, 6.5 mmol). The reaction mixture stirred at reflux while irradiating with a 250 W lamp. The reaction mixture was stirred at reflux for 3 h. The solution was cooled, filtered and the solvent evaporated in vacuo. The residue was subjected to column chromatography on silica gel with hexanes as the eluent to afford the title compound (0.570 g, 33% yield): 1H NMR (400 MHz, CDCl3) δ: 7.80 (1H, dd, J=7.8, 1.7 Hz), 7.49 (1H, d, J=5.4 Hz), 7.40-7.33 (3H, m), 4.78 (2H, s). LCMS (M+H)+ m/z 209.
Intermediate 139




Benzo[b]thiophen-7-ylmethanamine hydrochloride. To intermediate 138, 7-(bromomethyl)benzo[b]thiophene (0.20 g, 0.96 mmol) was added a methanolic solution saturated with ammonia (30 mL). The reaction mixture was heated in a steal bomb at 70° C. for 18 h. The solvent was evaporated in vacuo and the residue was dissolved in MeOH (10 mL). HCl (1M in ethanol, 1 mL) was added to the solution and the solvents were removed in vacuo to afford the title compound (0.177 g, 99% yield); LCMS (M+H)+ m/z 164.
Intermediates 140-148
The synthesis of intermediates 140-148 provides representative procedures for the synthesis of other compounds in the invention.
Intermediate 140




N-{4-Fluoro-2-(methylcarbamoyl)benzyl}-3-benzyloxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A mixture of intermediate 27, 3-benzyloxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid (0.172 g, 0.54 mmol) and intermediate 39, 2-(aminomethyl)-5-fluoro-N-methylbenzamide trifluoroacetate salt (0.177 g, 0.60 mmol) in dichloromethane (10 ml) was treated at 22° C. with triethylamine (0.17 ml, 1.22 mmol) followed by benzotriazole-1-yloxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PYBOP) (0.340 g, 0.65 mmol). After 3 h, the reaction mixture was diluted with ethyl acetate, washed with saturated sodium bicarbonate and brine then dried over anhydrous magnesium sulfate. Evaporation of the solvent followed by chromatography of the residue on silica gel (elution gradient ethyl acetate 50-100% in toluene) gave 0.260 g (100% yield) of the title amide as a white solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.70 (3H, d, J=6.6 Hz, CH3), 3.01 (3H, d, J=4.7 Hz, NCH3), 3.89 (2H, m, CH2), 4.14 (1H, m, CH), 4.29 (1H, m, CH), 4.53 (2H, d, J=6.7 Hz, NCH2), 4.69 (1H, q, J=6.6 Hz, OCH), 5.35 (2H, s, OCH2), 6.69 (1H, broad q, NH), 7.09 (1H, m, aromatic), 7.16 (1H, m, aromatic), 7.32 (3H, m, aromatics), 7.42 (1H, m, aromatic), 7.47 (2H, m, aromatics), 8.61 (1H, broad t, NH). HRMS (ESI+) calculated for C25H26FN4O5 [M+H+]: 481.1887: found: 481.1884.
Intermediate 141




N-(4-fluorobenzyl)-3-(benzyloxy)-9-(2-(methylthio)ethyl)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 22 (82 mg, 0.22 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) (152 mg, 0.4 mmol) in 1 mL dimethylformamide was stirred for 20 min under N2. 4-Fluorobenzylamine (38 mg, 0.3 mmol) was then added and stirring continued for 16 hrs. The dimethylformamide was evaporated under reduced pressure and the remaining residue dissolved in CH2Cl2. The resulting solution was washed with dil HCl. The solvent was removed under reduced pressure and the crude product purified by chromatography (SiO2, ethyl acetate) to provide the title compound (70 mg, Yield=66%). LC/MS m/e 484 (M+H).
Intermediate 142




3-(Benzyloxy)-2-(6-fluoro-1-oxoisoindoline-2-carbonyl)-9,9-dimethyl-6,7-dihydropyrimido[2,1-c][1,4]oxazin-4(9H)-one. To a stirred suspension of intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid, (1.65 g, 5 mmol) in CH2Cl2 (15 mL) was added a catalytic amount of dimethylformamide and 10 mL of a 2 M oxalylchloride in CH2Cl2. After 30 min., the resulting clear yellow solution was concentrated to afford a light brown solid. This solid was dissolved in CH2Cl2 (50 mL) and added to a stirred mixture of intermediate 107, 2-(aminomethyl)-5-fluorobenzonitrile trifluoroacetic acid salt, (1.77 g, 5.97 mmol) and diethylisopropylamine (2.6 mL, 15 mmol) in CH2Cl2 (100 mL). After 1 h, the clear brown reaction mixture was concentrated and the resulting residue dissolved in ethyl acetate (200 mL) then washed sequentially with water (25 mL), 1N HCl (25 mL), water (25 mL) and brine (25 mL). The combined aqueous layers were extracted with CH2Cl2 (3×50 mL). The combined organic phases were dried (Na2SO4), filtered, concentrated and purified by flash chromatography (SiO2), using 30-50% ethyl acetate/Hexanes as eluent to afford the title compound (0.331 g, 14% yield) as a yellow powder. 1H NMR (500 MHz, CDCl3) δ: 7.48-7.45 (1H, m), 7.88 (2H, d, J=7.9 Hz), 7.30 (2H, d, J=7.0 Hz), 7.12 (2H, t, J=7.6 Hz), 7.05-7.01 (1H, m), 5.22 (2H, s), 4.77 (2H, s), 4.06 (4H, s), 1.59 (6H, s). HRMS (M+H) calcd for C25H23FN3O5: 464.1622; found: 464.1628.
Intermediate 143




N-(4-Fluoro-2-(methylcarbamoyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A 25 mL round-bottom flask was charged with intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid, (0.991 g, 3.0 mmol), intermediate 39, 2-(aminomethyl)-5-fluoro-N-methylbenzamide trifluoroacetic acid salt (1.185 g, 4.0 mmol), 4-dimethylaminopyridine (DMAP, 1.1 g, 9.0 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU, 1.722 g, 4.5 mmol). Dimethylformamide (20 mL) was added and the mixture stirred for 1 h at ambient temperature. After this, the reaction mixture was diluted with ethyl acetate (100 mL) then washed with water (3×25 mL) and brine (25 mL). The organic layer was dried (Na2SO4), filtered, concentrated and purified by flash chromatography (SiO2), eluting with hexanes/ethyl acetate (1:1 to 1:3) followed by ethyl acetate, to afford the title compound as an off-white solid (1.48 g, 100% yield). 1H NMR (500 MHz, CDCl3) δ ppm: 8.49 (1H, t, J=6.1 Hz), 7.48-7.46 (2H, m), 7.43 (1H, dd, J=8.5, 5.5 Hz), 7.31-7.27 (3H, m), 7.12 (1H, dd, J=8.9, 2.7 Hz), 7.05 (1H, td, J=8.2, 2.7 Hz), 6.52 (1H, br s), 5.26 (2H, s), 4.53 (2H, d, J=6.4 Hz), 4.01 (2H, t, J=4.9 Hz), 3.96 (2H, t, J=4.9 Hz), 2.97 (3H, d, J=4.9 Hz), 1.62 (6H, s). HRMS (M+H) calcd for C26H28FN4O5: 495.2044; found: 495.2032.
Intermediate 144




N-(4-Fluoro-2-hydroxybenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid, (1.50 g, 4.54 mmol) and HATU (2.07 g, 5.45 mmol) in anhydrous dimethylformamide was stirred for 20 minutes under a nitrogen atmosphere at room temperature. To the solution was added intermediate 135, 2-hydroxy-4-fluoro-benzylamine hydrochloride, (1.05 g, 5.9 mmol) followed by DMAP (1.39 g, 11.4 mmol), and the reaction mixture stirred at 60° C. for 90 minutes. The solvent was removed in vacuo and the crude residue purified by flash column chromatography (SiO2), eluting with 40%-60% ethyl acetate in hexanes to give the title compound (1.31 g, 64% yield) as a solid. 1H NMR (500 MHz, CDCl3) δ ppm: 9.69 (1H, br s), 8.18 (1H, t, J=6.3 Hz), 7.44 (2H, dd, J=6.6, 2.6 Hz), 7.30-7.35 (3H, m), 7.00 (1H, dd, J=8.2, 6.7 Hz), 6.68 (1H, dd, J=110.4, 2.4 Hz), 6.54 (1H, dt, J=8.2, 2.4 Hz), 5.29 (2H, s), 4.36 (2H, d, J=6.7 Hz), 3.97-4.05 (4H, m), 1.61 (6H, s).
Intermediate 145




N-(4-Fluoro-2-(2-oxopiperidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazin-4(9H)-one. To intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid, (80 mg, 0.242 mmol) in CH3CN/dimethylformamide (5 mL: 1 mL:) was added intermediate 125, 1-(2-(aminomethyl)-5-fluorophenyl)piperidin-2-one hydrochloride salt, (69 mg, 0.266 mmol), benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP) (139 mg, 0.266 mmol) and diisopropylethylamine (169 μL, 0.968 mmol). The reaction mixture was stirred at 23° C. for 3 h. The solvents were then removed in vacuo and an aqueous solution of HCl (1N, 25 mL) was added. This was extracted with ethyl acetate (3×25 mL). The combined organic fractions were dried (MgSO4), filtered and concentrated in vacuo. The crude material was purified on a Biotage system using a silica gel column with Hexanes/ethyl acetate (1:1) to (1:5) as eluent to afford 114 mg (88% yield) of the title compound. 1HNMR 400 MHz (DMSO) δ ppm: 8.73 (1H, dd, (t), J=6.0 Hz), 7.46 (2H, m), 7.35 (4H, m), 7.18 (1H, dd, J=9.8, 3.0 Hz) 7.0 (1H, m), 5.10 (2H, s), 4.43 (1H, dd, J=15.0, 7.08 Hz), 4.06 (1H, dd, J=15.0, 5.56), 4.02 (2H, t, J=5.0 Hz), 3.90 (2H, t, J=5.0 Hz), 3.64 (1H, m), 3.44 (1H, m), 2.45 (1H, m), 2.35 (1H, m), 1.86 (4H, m), 1.56 (6H, s). LCMS (M+H)+ m/z 535.
Intermediate 146




N-(4-Fluoro-2-(2-oxoazetidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid, (0.142 g, 0.430 mmol) in CH3CN: dimethylformamide (30 mL: 5 mL) was added intermediate 120, 1-(2-(aminomethyl)-5-fluorophenyl)azetidin-2-one hydrochloride, (0.100 g, 0.43 mmol), (benzotriazole-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (0.207 g, 0.470 mmol) and diisopropylethylamine (280 μL, 1.72 mmol). The reaction mixture was stirred at 23° C. for 18 h. The solvents were removed in vacuo and 1N HCl (50 mL) added. This was extracted with ethyl acetate (2×50 mL). The combined organic extracts were dried (MgSO4), filtered and concentrated in vacuo. The residue was purified on a Biotage system using a silica gel column with ethyl acetate as eluent to afford the title compound (0.157 g, 72% yield). 1H NMR (400 MHz, MeOD) δ ppm: 7.88 (1H, d, J=8.3 Hz), 7.76 (1H, d, J=8.0 Hz), 7.58-7.30 (4H, m), 7.10 (1H, dd, J=10.2, 2.5 Hz), 6.90 (1H, m), 5.20 (2H, s), 4.55 (2H, s), 4.04 (2H, m), 3.96 (2H, m), 3.78 (2H, t, J=4.2 Hz), 3.10 (2H, t, J=4.2 Hz), 1.58 (6H, s); LCMS (M+H)+ m/z 506.
Intermediate 147




N-(Benzo[b]thiophen-7-ylmethyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid (0.200 g, 0.60 mmol) in CH3CN:dimethylformamide (30 mL: 5 mL) was added intermediate 139, benzo[b]thiophen-7-ylmethanamine hydrochloride (0.069 g, 0.266 mmol), benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PYBOP) (0.347 g, 0.670 mmol) and diisopropylethylamine (420 μL, 2.40 mmol). The reaction mixture was stirred at 23° C. for 60 h. The solvents were then removed in vacuo and 1N HCl (50 mL) added. This was extracted with ethyl acetate (2×50 mL). The combined organic extracts were dried (MgSO4), filtered and concentrated in vacuo. The residue was purified on a Biotage system using a silica gel column with hexane:ethyl acetate (1:1) to (1:5) gradient as eluent to afford the title compound (0.279 g, 87% yield): 1H NMR (400 MHz, DMSO-d6) δ: 9.11 (1H, dd, J=5.6 Hz), 7.81 (1H, m), 7.77 (1H, d, J=5.6 Hz), 7.52 (1H, d, J=5.6 Hz), 7.48-7.26 (6H, m), 5.09 (2H, s), 4.67 (2H, d, J=5.2 Hz), 4.04 (2H, m), 3.90 (2H, m), 1.58 (6H, s). LCMS (M+H)+ m/z 476.
Intermediate 148




N-(4-Fluoro-2-(methylcarbamoyl)benzyl)-3-(benzyloxy)-10,10-dimethyl-4-oxo-6,7,8,10-tetrahydro-4H-pyrimido[2,1-c][1,4]oxazepine-2-carboxamide. A solution of intermediate 35, 3-(benzyloxy)-10,10-dimethyl-4-oxo-6,7,8,10-tetrahydro-4H-pyrimido[2,1-c][1,4]oxazepine-2-carboxylic acid in anhydrous dimethylformamide (4 mL) was treated with HATU (0.278 g, 0.70 mmol) and stirred for 10 minutes. The reaction mixture was treated with 2-(aminomethyl)-5-fluoro-N-methylbenzamide trifluoroacetic acid salt (0.24 g, 0.8 mmol), followed by dimethylaminopyridine (0.121 g, 0.97 mmol), then stirred for 4 hours at 60° C. Following this, the solvent was removed in vacuo and the remaining residue dissolved in ethyl acetate (15 mL) and washed with 1.0 N HCl (15 mL). The organic layer was dried (sodium sulfate), filtered, and concentrated to dryness. The crude product was purified by flash column chromatography (SiO2), eluting with ethyl acetate. Fractions containing the product were pooled, concentrated to dryness and triturated with ether to provide 0.366 g of the title compound as a white glassy solid. 1H NMR (500 MHz, d6-acetone) δ ppm: 8.71-8.82 (1H, m), 7.84-7.95 (1H, br), 7.47-7.62 (4H, m), 7.27-7.40 (5H, m), 7.20 (1H, dt, J=8.5, 2.7 Hz), 5.15-5.21 (2H, br s), 4.48-4.60 (4H, m), 2.96 (2H, s), 2.94 (2H, s), 1.62-1.64 (6H, s).
Intermediates 149-174
Intermediates 149-174 were prepared according to the coupling procedures described for intermediates 140-148.
Intermediate 149




N-(4-Fluorobenzyl)-3-(benzyloxy)-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 16, 3-benzyloxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid and 4-fluorobenzylamine. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.67 (3H, d, J=6.6 Hz, CH3), 3.91 (2H, m, CH2), 4.12-4.35 (2H, m, CH2), 4.52 (2H, d, J=5.9 Hz, NCH2), 4.70 (1H, q, J=6.6 Hz, OCH), 5.33 (2H, s, OCH2), 7.02 (2H, m, aromatics), 7.25 (2H, m, aromatics), 7.35 (3H, m, aromatics), 7.47 (2H, m, aromatics), 7.71 (1H, broad t, NH).
Intermediate 150




N-(4-Fluoro-2-(methylcarbamoyl)benzyl)-3-(benzyloxy)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 7,3-(benzyloxy)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 39, 2-(aminomethyl)-5-fluoro-N-methylbenzamide. White crystals; mp 189-190° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ (ppm): 3.01 (3H, d, J=4.5 Hz, NCH3), 4.00 (2H, m, CH2), 4.08 (1H, m, CH), 4.50 (2H, d, J=6.6 Hz, NCH2), 4.71 (2H, s, OCH2), 5.38 (2H, s, OCH2), 6.88 (1H, broad q, NH), 7.07 (1H, m, aromatic), 7.16 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.30-7.44 (6H, m, aromatics), 8.55 (1H, broad t, NH). Anal. Calcd for C24H23FN4O5: C, 61.80; H, 4.97; N, 12.01. Found: C, 61.84; H, 4.82; N, 12.00.
Intermediate 151




N-(4-Fluoro-2-(1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from 3-(benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid which was synthesized using the method described for the synthesis or intermediates 7 and 16, and intermediate 69, (4-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl)methanamine. White needles; mp 155-157° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.03 (3H, t, J=7.5 Hz, CH3), 1.97-2.02 (1H, m, CH), 2.29-2.32 (1H, m, CH), 3.83-3.88 (2H, m, CH2), 4.15-4.31 (2H, m, CH2), 4.44 (2H, m, CH2), 4.53 (1H, m, CH), 5.34 (2H, s, OCH2), 7.08 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.20 (1H, m, aromatic), 7.29-7.31 (3H, m, aromatics), 7.47 (2H, m, aromatics), 7.74 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 8.02 (1H, s, CH), 8.41 (1H, s, CH), 8.55 (1H, broad t, NH). Anal. Calcd for C26H25FN6O4: C, 61.89; H, 4.99; N, 16.65. Found: C, 61.67; H, 5.13; N, 16.61.
Intermediate 152




N-(4-Fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 85, (4-fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl)methanamine. White crystals; mp 203° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.65 (6H, s, 2×CH3), 2.50 (3H, s, CH3), 4.03 (4H, m, 2×CH2), 4.46 (2H, d, J=6.6 Hz, NCH2), 5.31 (2H, s, OCH2), 7.06 (1H, dd, J=3 Hz and J=8.6 Hz, aromatic), 7.16 (1H, m, aromatic), 7.30-7.34 (3H, m, aromatics), 7.50 (2H, m, aromatics), 7.74 (1H, dd, J=6.0 Hz and J=8.6 Hz, aromatic), 8.28 (1H, s, CH), 8.45 (1H, broad t, NH). Anal. Calcd for C27H27FN6O4: C, 62.54; H, 5.25; N, 16.21. Found: C, 62.48; H, 5.31; N, 16.29.
Intermediate 153




N-(2-Fluoro-4-(3-methyl-1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 87, (2-fluoro-4-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl)methanamine. White crystals; mp 183-185° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.65 (6H, s, 2×CH3), 2.52 (3H, s, CH3), 4.05 (4H, m, 2×CH2), 4.64 (2H, d, J=6.1 Hz, NCH2), 5.33 (2H, s, OCH2), 7.29-7.55 (8H, m, aromatics), 7.84 (1H, broad t, NH), 8.45 (1H, s, CH). Anal. Calcd for C27H27FN6O4: C, 62.54; H, 5.25; N, 16.21. Found: C, 62.41; H, 5.40; N, 16.23.
Intermediate 154




Methyl 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorobenzoate. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 38, methyl 2-(aminomethyl)-5-fluorobenzoate. 1H NMR (CDCl3, 500 MHz) δ ppm: 1.61 (6H, s, gem-Me), 3.88 (3H, s, OMe), 3.97 (2H, t, J=5.5 Hz, CH2), 4.02 (2H, t, J=5.5 Hz, CH2), 4.73 (2H, d, J=6.7 Hz, NCH2), 5.25 (2H, s, OCH2), 7.19 (1H, dt, J=3, 8.5 Hz, Ar—H), 7.27-7.31 (3H, m, Ar—Hs), 7.48-7.50 (2H, m, Ar—Hs), 7.61 (1H, dd, J=5.5, 8.5 Hz, Ar—H), 7.66 (1H, dd, J=3, 9.5 Hz, Ar—H). LC/MS m/z 496 (M+H).
Intermediate 155




N-(2-(Cyclopropylcarbamoyl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 40, 2-(aminomethyl)-N-cyclopropyl-5-fluorobenzamide. LC/MS m/z 521 (M+H).
Intermediate 156




N-(4-Fluoro-2-(morpholine-4-carbonyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 44, (2-(aminomethyl)-5-fluorophenyl)(morpholino)methanone. 1H NMR (CDCl3, 500 MHz) δ ppm: 1.59 (6H, s), 3.29 (2H, brs), 3.57 (2H, m), 3.74 (4H, s), 3.98 (4H, m), 5.26 (2H, s), 6.88 (1H, dd, J=8.2, 2.7 Hz), 7.03 (1H, dt, J=8.5, 2.5 Hz), 7.24-7.33 (3H, m), 7.42 (2H, dd, J=8.6, 5.3 Hz), 7.47 (2H, dd, J=7.5, 2.0 Hz), 8.18 (1H, t, J=6.4 Hz); LC/MS m/z 551 (M+H).
Intermediate 157




N-(4-Fluoro-2-(2-morpholino-2-oxoethoxy)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][, 4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 131, 2-(2-(aminomethyl)-5-fluorophenoxy)-1-morpholinoethanone. 1H NMR (CDCl3, 500 MHz) δ ppm: 1.59 (6H, s, gem-Me), 3.38, 3.54 (4H, br, NCH2), 3.62 (4H, m, OCH2), 3.96 (2H, m, NCH2), 4.01 (2H, m, OCH2), 4.55 (2H, s, OCH2), 4.55 (2H, d, J=4.3 Hz, NCH2), 5.17 (2H, s, OCH2), 6.53 (1H, dd, J=10, 2.1 Hz, Ar—H), 6.63 (1H, dt, J=2.5, 8 Hz, Ar—H), 7.23-7.26 (1H, m, Ar—H), 7.28-7.30 (3H, m, Ar—Hs), 7.42-7.44 (2H, m, Ar—Hs), 8.00 (1H, t, J=5.5 Hz, NH); 13C NMR (CDCl3, 125.77 Hz) δ ppm: 27.74, 38.76, 42.30, 42.98, 45.38, 58.06, 66.55, 66.78, 66.91, 74.72, 76.27, 100.40, 100.61, 108.07, 108.23, 122.56, 128.37, 128.49, 128.84, 130.88, 130.96, 139.69, 141.36, 141.98, 156.70, 157.02, 157.10, 159.58, 162.04, 163.99, 162.86, 165.79. HRMS (ESI) calcd for C30H34N4O7F (M+H) 581.2412, found 581.2393.
Intermediate 158




Dimethyl-carbamic acid 2-{[(3-benzyloxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydro-pyrimido[2,1-c][1,4]oxazine-2-carbonyl)-amino]-methyl}-5-fluoro-phenyl ester. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and intermediate 133, 2-(aminomethyl)-5-fluorophenyl dimethylcarbamate. 1H NMR (CDCl3, 500 MHz) δ ppm: 1.58 (6H, s, gem-Me), 2.92, 3.05 (2s, NMe), 3.96 (2H, m, NCH2), 4.00 (2H, m, OCH2), 4.48 (2H, d, J=5.5 Hz, NCH2), 5.26 (2H, s, OCH2), 6.84 (1H, dd, J=2.5 Hz, 9 Hz, Ar—H), 6.87 (1H, dt, J=2.5 Hz, 8 Hz, Ar—H), 7.25-7.33 (4H, m, Ar—Hs), 7.53 (2H, d, J= ̃7 Hz, Ar—Hs), 7.78 (1H, brt, J=5 Hz, NH); 13C NMR (CDCl3, 125.77 Hz) δ ppm: 27.68, 36.61, 36.89, 37.97, 42.97, 58.06, 74.73, 76.23, 110.58, 110.77, 113.06, 113.23, 126.55, 126.58, 128.31, 128.45, 128.91, 131.23, 131.31, 136.76, 140.70, 142.00, 150.60, 150.69, 154.50, 156.30, 159.79, 161.40, 163.37, 162.43. HRMS (ESI) calcd for C27H30N4O6F (M+H) 525.2149, found 525.2163.
Intermediate 159




N-(4-Fluoro-2-(2-oxopyrrolidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and 1-(2-(aminomethyl)-5-fluorophenyl)pyrrolidin-2-one, derived from reduction of intermediate 111, 4-fluoro-2-(2-oxopyrrolidin-1-yl)benzonitrile. 1HNMR 400 MHz (MeOD) δ ppm: 7.44 (3H, m), 7.33 (3H, m), 7.11 (1H, dd, J=9.2, 3.0 Hz) 7.03 (1H, m), 5.21 (2H, s), 4.43 (2H, s), 4.08 (2H, t, J=5.0 Hz), 3.98 (2H, t, J=5.0 Hz), 3.85 (2H, t, J=7.1 Hz), 2.58 (2H, t, J=8.0 Hz), 2.23 (2H, m), 1.61 (6H, s). LCMS (M+H)+ m/z 521.
Intermediate 160




N-(4-Fluoro-2-(2-oxoazepan-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2 carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and 1-(2-(aminomethyl)-5-fluorophenyl)azepan-2-one, derived from reduction of intermediate 113, 4-fluoro-2-(2-oxoazepan-1-yl)benzonitrile. 1HNMR 400 MHz (DMSO) δ ppm: 8.80 (1H, dd, (t), J=6.0 Hz), 7.46 (2H, m), 7.36 (4H, m), 7.08 (1H, dd, J=9.8, 2.8 Hz), 7.0 (1H, m), 5.09 (2H, s), 4.43 (1H, dd, J=15.2, 7.1 Hz), 4.06 (1H, dd, J=15.2, 5.0 Hz), 4.02 (2H, t, J=5.0 Hz), 3.90 (2H, t, J=5.0 Hz), 3.77 (1H, m), 3.51 (1H, m), 2.70 (1H, m), 2.51 (2H, m), 1.76 (6H, m), 1.56 (6H, s). LCMS (M+H)+ m/z 549.
Intermediate 161




N-(2-(2-Oxooxazolidin-3-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and 3-(2-(aminomethyl)phenyl)oxazolidin-2-one, derived from reduction of intermediate 116, 2-(2-oxooxazolidin-3-yl)benzonitrile. 1H NMR (400 MHz, DMSO-d6) δ ppm: 8.90 (1H, t, J=6.0 Hz), 7.58-7.32 (7H, m), 7.22 (2H, t, J=7.5 Hz), 5.08 (2H, s), 4.48 (2H, t, J=7.8 Hz), 4.44 (2H, d, J=6.0 Hz), 4.06-3.97 (4H, m), 3.88 (2H, m), 1.56 (6H, s); LCMS (M+H)+ m/z 505.
Intermediate 162




N-(2-(2-Oxoazetidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and 1-(2-(aminomethyl)phenyl)azetidin-2-one, derived from reduction of intermediate 115, 2-(2-oxoazetidin-1-yl)benzonitrile. 1H NMR (400 MHz, CDCl3) δ ppm: 8.67 (1H, brt, J=6.3 Hz), 7.60 (1H, dd, J=1.3, 7.6 Hz), 7.53-7.50 (1H, dd, m), 7.34-7.24 (5H, m), 7.18 (1H, ddd(dt), J=1.2, 7.4 Hz), 7.10 (1H, dd, J=1.2, 8.0 Hz), 5.27 (2H, s), 4.60 (2H, d, J=6.3 Hz), 4.01-3.95 (4H, m), 3.71 (2H, t, J=4.5 Hz), 3.10 (2H, t, J=4.5 Hz), 1.60 (6H, s); LCMS (+ESI, M+H+) m/z 489.
Intermediate 163




N-(4-Fluoro-2-(thiazol-2-ylamino)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and N-(2-(aminomethyl)-5-fluorophenyl)thiazol-2-amine, derived from reduction of intermediate 123, 4-fluoro-2-(thiazol-2-ylamino)benzonitrile. 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.92 (1H, s), 9.10 (1H, t, J=6.3 Hz), 8.19 (1H, dd, J=2.8, 12.3 Hz), 7.41-7.38 (2H, m), 7.32-7.28 (5H, m), 6.99 (1H, d, J=3.8 Hz), 6.71 (1H, ddd(dt), J=2.8, 8.3), 5.05 (2H, s), 4.45 (2H, d, J=6.3 Hz), 4.01-3.98 (2H, m), 3.88-3.85 (2H, m), 1.54 (6H, s) LCMS (+ESI, M+H+) m/z 536.
Intermediate 164




N-(4-Fluoro-2-(5-methyl-1,3,4-thiadiazol-2-ylamino)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and N-(2-(aminomethyl)-5-fluorophenyl)-5-methyl-1,3,4-thiadiazol-2-amine, derived from reduction of intermediate 124, 4-fluoro-2-(5-methyl-1,3,4-thiadiazol-2-ylamino)benzonitrile. LCMS (+ESI, M+H+) m/z 551.
Intermediate 165




N-(4-Fluoro-2-(2-oxooxazolidin-3-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and 3-(2-(aminomethyl)-5-fluorophenyl)oxazolidin-2-one, derived from reduction of intermediate 117, 4-fluoro-2-(2-oxooxazolidin-3-yl)benzonitrile. 1H NMR (400 MHz, MeOD) δ ppm: 9.48 (1H, dd, J=8.6, 6.5 Hz), 7.41 (2H, m), 7.32 (3H, m), 7.22 (1H, dd, J=9.6, 2.5 Hz), 7.08 (1H, td, J=8.6, 2.7 Hz), 5.21 (2H, s), 4.57 (2H, t, J=7.7 Hz), 4.50 (2H, s), 4.07 (4H, m), 3.99 (2H, m), 1.61 (6H, s). LCMS (M+H)+ m/z 523.
Intermediate 166




(R)—N-(2-(2-((tert-Butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and (S)-1-(2-(aminomethyl)-5-fluorophenyl)-5-((tert-butyldimethylsilyloxy)methyl)pyrrolidin-2-one, derived from reduction of intermediate 122, (R)-2-(2-((tert-butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzonitrile 1HNMR 400 MHz (DMSO) δ ppm: 8.82 (1H, broad s), 7.47 (2H, m), 7.35 (4H, m), 7.00 (1H, broad s), 5.09 (2H, s), 4.60-4.10 (3H, m), 4.02 (3H, m), 3.88 (2H, m), 3.53 (2H, broad s), 2.42-2.35 (2H, m), 1.94 (1H, m), 1.56 (3H, s), 1.55 (3H, s), 0.82 (9H, broad s), −0.01 (6H, s). LCMS (M+H)+ m/z 665.
Intermediate 167




(S)—N-(2-(2-((tert-Butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and (S)-1-(2-(aminomethyl)-5-fluorophenyl)-5-((tert-butyldimethylsilyloxy)methyl)pyrrolidin-2-one, derived from reduction of intermediate 121, (R)-2-(2-((tert-butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzonitrile. 1HNMR 400 MHz (DMSO) δ ppm: 8.82 (1H, broad s), 7.52-7.33 (7H, m), 7.02 (1H, broad s), 5.60 (2H, s), 4.60-4.20 (2H, m), 4.02 (3H, t, J=5.0 Hz), 3.89 (3H, t, 5.0 Hz), 3.53 (2H, broad s), 2.51 (2H, s), 2.44-2.26 (2H, m), 1.94 (1H, broad s), 1.57 (3H, s), 1.55 (3H, s), 0.82 (9H, broad s), −0.02 (6H, s). LCMS (M+H)+ m/z 665.
Intermediate 168




N-(4-Fluoro-2-(N-methylacetamido)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid N-(2-(aminomethyl)-5-fluorophenyl)-N-methylacetamide, derived from reduction of intermediate 114, N-(2-cyano-5-fluorophenyl)-N-methylacetamide. 1HNMR 400 MHz (MeOD) δ ppm: 7.49-7.33 (6H, m), 7.15-7.06 (2H, m), 5.22 (2H, s), 4.41 (2H, d, J=2.4 Hz), 4.09 (2H, t, J=5.1 Hz), 4.01 (2H, t, J=5.1 Hz), 3.25 (0.4H, s), 3.22 (2.6H, s), 1.85 (3H, s), 1.63 (6H, s). LCMS (M+H)+ m/z 509.
Intermediate 169




N-(2-Amino-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid 2-(aminomethyl)-5-fluorobenzenamine. 1HNMR 400 MHz (MeOD) δ ppm: 1HNMR 400 MHz (MeOD) δ ppm: 7.36 (2H, m), 7.30 (3H, m), 7.10 (1H, dd, J=8.3, 6.5 Hz), 6.47 (1H, dd, J=11.1, 2.5 Hz), 6.32 (1H, ddd, (dt), J=8.6, 2.5 Hz), 5.19 (2H, s), 4.40 (2H, s), 4.07 (2H, t, J=5.0 Hz), 3.99 (2H, t, J=5.0 Hz), 1.61 (6H, s). LCMS (M+H)+ m/z 453.
Intermediate 170




N-(2-(Ethylamino)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and 2-(aminomethyl)-N-ethyl-5-fluorobenzenamine. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.27 (3H, t, J=7.1 Hz, CH3), 1.63 (6H, s, 2×CH3), 3.10 (2H, q, J=7.1 Hz, CH2), 4.04 (4H, m, 2×CH2), 4.45 (2H, d, J=6.6 Hz, NCH2), 5.27 (2H, s, OCH2), 6.28 (1H, broad s, aromatic), 6.31 (1H, m, aromatic), 6.98 (1H, m, aromatic), 7.3-7.38 (3H, m, aromatics), 7.49 (2H, m, aromatics), 7.54 (1H, broad t, NH). HRMS (ESI+) calculated for C26H30FN4O4 [M+H+]: 481.2251: found: 481.2254.
Intermediate 171




N-(4-Fluorobenzyl)-3-(benzyloxy)-N-methoxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and (4-fluorophenyl)-N-methoxymethanamine. White crystals; mp 141° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.66 (6H, s, 2×CH3), 3.59 (3H, s, OCH3), 4.07 (4H, m, 2×CH2), 4.90 (2H, s, NCH2), 5.20 (2H, s, OCH2), 6.81 (2H, m, aromatics), 7.26-7.30 (3H, m, aromatics), 7.36 (2H, m, aromatics), 7.44 (2H, m, aromatics). HRMS (ESI+) calculated for C25H27FN3O5 [M+H+]: 468.1935: found: 468.1916.
Intermediate 172




N-(4-Fluoro-2-(1,2,3-thiadiazol-4-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and (4-fluoro-2-(1,2,3-thiadiazol-4-yl)phenyl)methanamine. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.66 (6H, s, 2×CH3), 4.02 (4H, m, 2×CH2), 4.57 (2H, d, J=6.6 Hz, NCH2), 5.32 (2H, s, OCH2), 7.18 (1H, m, aromatic), 7.27-7.34 (4H, m, aromatics), 7.54 (2H, m, aromatics), 7.74 (1H, dd, J=6.2 Hz and J=8.6 Hz, aromatic), 8.71 (1H, s, CH), 8.80 (1H, broad t, NH). HRMS (ESI+) calculated for C26H25FN5O4S [M+H+]: 522.1611: found: 522.1601.
Intermediate 173




N-(4-Fluoro-2-(5-methyloxazol-2-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and (4-fluoro-2-(5-methyloxazol-2-yl)phenyl)methanamine. White crystals; mp 186° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.61 (6H, s, 2×CH3), 2.43 (3H, s, CH3), 4.02 (4H, m, 2×CH2), 4.80 (2H, d, J=6.3 Hz, NCH2), 5.25 (2H, s, OCH2), 6.82 (1H, s, CH), 7.11 (1H, m, aromatic), 7.29-7.34 (3H, m, aromatics), 7.52 (2H, m, aromatics), 7.65 (1H, dd, J=2.5 Hz and J=9.6 Hz, aromatic), 7.69 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 9.32 (1H, broad t, NH). HRMS (ESI+) calculated for C28H28FN4O5 [M+H+]: 519.2044: found: 519.2024.
Intermediate 174




N-(4-Fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 27, 3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido-[2,1-c][1,4]oxazine-2-carboxylic acid and (4-fluoro-2-iodophenyl)methanamine. White solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.66 (6H, s, 2×CH3), 4.04 (4H, m, 2×CH2,), 4.57 (2H, d, J=6.6 Hz, NCH2), 7.05 (1H, m, aromatic), 7.3-7.38 (3H, m, aromatics), 7.42 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 7.53 (2H, m, aromatics), 7.56 (1H, dd, J=2.6 Hz and J=8.0 Hz, aromatic), 8.05 (1H, broad t, NH). HRMS (ESI+) calculated for C24H24FIN3O4 [M+H+]: 564.0796; found: 564.0809.
Intermediate 175




N-(4-Fluoro-2-(2-methoxypyridin-3-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. Intermediate 174, N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.350 g, 0.62 mmol) in a mixture of acetonitrile (12 ml) and water (12 ml) was treated with 2-methoxypyridin-3-ylboronic acid (0.190 g, 1.24 mmol), sodium carbonate (0.20 g, 1.88 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.15 g). The reaction mixture was degassed, flushed with argon and heated at 90° C. for 30 min. The reaction mixture was then diluted with ethyl acetate, washed with water and brine, dried over anhydrous magnesium sulfate and concentrated. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in hexane) gave 0.245 g (72% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.63 (6H, s, 2×CH3), 3.89 (3H, s, OCH3), 4.04 (4H, m, 2×CH2), 4.37 (2H, broad, NCH2), 5.26 (2H, s, OCH2), 6.93 (1H, dd, J=2.5 Hz and J=9 Hz, aromatic), 7.0 (1H, dd, J=5 Hz and J=7 Hz, aromatic), 7.06 (1H, m, aromatic), 7.3-7.5 (8H, m, aromatics and NH), 8.24 (1H, m, aromatic). HRMS (ESI+) calculated for C30H30FN4O5 [M+H+]: 545.2200; found: 545.2184.
Intermediate 176




N-(4-Fluoro-2-phenyl-benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. Intermediate 174, N-(4-Fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.150 g, 0.266 mmol) in a mixture of acetonitrile (10 ml) and water (10 ml) was treated with phenylboronic acid (0.042 g, 0.35 mmol), sodium carbonate (0.062 g, 0.58 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.070 g). The reaction mixture was degassed, flushed with argon and heated at 90° C. for 30 min. The reaction mixture was then diluted with ethyl acetate, washed with water and brine, dried over anhydrous magnesium sulfate and concentrated. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in hexane) gave 0.124 g (91% yield) of the title material as a light yellow solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.61 (6H, s, 2×CH3), 4.03 (4H, m, 2×CH2), 4.49 (2H, d, J=6.1 Hz, NCH2), 7.02 (2H, m, aromatics), 7.29-7.51 (12H, m, aromatics and NH). HRMS (ESI+) calculated for C30H29FN3O4 [M+H+]: 514.2142; found: 514.2137.
Intermediate 177




N-(4-Fluoro-2-(1H-pyrazol-5-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. Intermediate 174, N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.150 g, 0.27 mmol) was reacted with 1H-pyrazol-5-ylboronic acid (0.060 g, 0.54 mmol), sodium carbonate (0.085 g, 0.81 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.070 g) to give 0.085 g (62% yield) of the title material as a white solid after chromatography on silica gel. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.63 (6H, s, 2×CH3), 4.03 (4H, m, 2×CH2), 4.65 (2H, d, J=6.5 Hz, NCH2), 5.25 (2H, s, OCH2), 6.54 (1H, d, J=2.5 Hz, CH), 7.03 (1H, m, aromatic), 7.25 (1H, dd, J=2.5 Hz and J=9.8 Hz, aromatic), 7.35 (3H, m, aromatics), 7.53 (2H, m, aromatics), 7.56 (1H, d, J=2.5 Hz, CH), 7.60 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 8.96 (1H, broad t, NH). HRMS (ESI+) calculated for C27H27FN5O4 [M+H+]: 504.2047: found: 504.2068.
Intermediate 178




N-(4-Fluoro-2-(2-(trimethylsilyl)ethynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][, 4]oxazine-2-carboxamide. A solution of intermediate 174, N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.277 g, 0.49 mmol) in a mixture of N,N-dimethylformamide (3 ml) and piperidine (1.2 ml) was treated under argon with dichlorobis(triphenylphosphine)palladium(II) (0.020 g), triphenylphosphine (0.010 g), copper(1) iodide (0.010 g) followed by (trimethylsilyl)acetylene (0.21 ml, 1.47 mmol). The resulting mixture was sealed and heated at 50° C. for one hour. The reaction mixture was then diluted with ethyl acetate, washed with water and brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in hexane) gave 0.164 g (63% yield) of the title material as a light yellow solid. 1HNMR 400 MHz (CDCl3) δ ppm: 0.27 (9H, s, SiCH3), 1.64 (6H, s, 2×CH3), 4.04 (4H, m, 2×CH2), 4.71 (2H, d, J=6.1 Hz, NCH2), 5.32 (2H, s, OCH2), 6.99 (1H, m, aromatic), 7.19 (1H, dd, J=2.6 Hz and J=9.1 Hz, aromatic), 7.3-7.37 (4H, m, aromatics), 7.48-7.51 (2H, m, aromatics), 7.75 (1H, broad t, NH). HRMS (ESI+) calculated for C29H33FN3O4Si [M+H+]: 534.2224; found: 534.2229.
Intermediate 179




N-(2-Ethynyl-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 178, N-(4-fluoro-2-(2-(trimethylsilyl)ethynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.150 g, 0.28 mmol) in methanol (5 ml) was treated with potassium carbonate (0.120 g, 0.84 mmol) and the resulting mixture stirred at 22° C. for one hour. The reaction mixture was then diluted with ethyl acetate, washed with water and brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give 0.129 g (100% yield) of the title material as a light yellow solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.64 (6H, s, 2×CH3), 3.31 (1H, s, CH), 4.04 (4H, m, 2×CH2), 4.69 (2H, d, J=6.6 Hz, NCH2), 5.31 (2H, s, OCH2), 7.04 (1H, m, aromatic), 7.21 (1H, dd, J=2.6 Hz and J=9.1 Hz, aromatic), 7.32-7.42 (3H, m, aromatics), 7.40 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 7.52-7.54 (2H, m, aromatics), 8.00 (1H, broad t, NH). HRMS (ESI+) calculated for C26H25FN3O4 [M+H]: 462.1829; found: 462.1822.
Intermediate 180




N-(4-Fluoro-2-(3-methylisoxazol-5-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 179, N-(2-ethynyl-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.150 g, 0.32 mmol) in dimethyl sulfoxide (5 ml) was treated with nitromethane (0.14 ml, 1.92 mmol), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM) (0.241 g, 1.0 mmol) (M. Kunishima et al., Tetrahedron, 55, 1999, 13159-13170) and 4-(dimethylamino)pyridine (DMAP) (0.010 g) and the resulting mixture stirred at 22° C. for 16 h. Identical quantities of nitromethane, DMTMM and DMAP were then added and the mixture was stirred for another 24 h. The reaction mixture was diluted with ethyl acetate, washed with saturated sodium bicarbonate and brine, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in dichloromethane) gave 0.122 g (73% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.62 (6H, s, 2×CH3), 2.39 (3H, s, CH3), 4.04 (4H, m, 2×CH2), 4.69 (2H, d, J=6.1 Hz, NCH2), 5.31 (2H, s, OCH2), 6.36 (1H, m, CH), 7.13 (1H, m, aromatic), 7.30-7.35 (4H, m, aromatics), 7.50-7.53 (2H, m, aromatics), 7.58 (1H, dd, J=5.5 Hz and J=8.6 Hz, aromatic), 8.06 (1H, broad t, NH). HRMS (ESI+) calculated for C28H28FN4O5 [M+H+]: 519.2044; found: 519.2059.
Intermediate 181




N-(2-(3-Bromoisoxazol-5-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 179, N-(2-ethynyl-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.350 g, 0.76 mmol) in a mixture of ethyl acetate (10 ml) and water (2 ml) was treated with potassium bicarbonate (0.230 g, 2.3 mmol) followed by dibromoformaldoxime (0.354 g, 1.75 mmol) (D. M. Vyas, Y. Chiang and T. W. Doyle, Tetrahedron Letters, 1984, 25, 487-490) and the resulting mixture stirred at 22° C. After 1 h, identical quantities of potassium bicarbonate and dibromoformaldoxime were added and the mixture stirred for another 1.5 h. The reaction mixture was then diluted with ethyl acetate, washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Crystallization of the residue from ether gave 0.300 g (68% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.63 (6H, s, 2×CH3), 4.04 (4H, m, 2×CH2), 4.67 (2H, d, J=6.6 Hz, NCH2), 5.32 (2H, s, OCH2), 6.60 (1H, s, CH), 7.17 (1H, m, aromatic), 7.30-7.36 (4H, m, aromatics), 7.51-7.53 (2H, m, aromatics), 7.60 (1H, dd, J=5.5 Hz and J=8.6 Hz, aromatic), 8.01 (1H, broad t, NH). HRMS (ESI+) calculated for C27H25BrFN4O5 [M+H+]: 583.0992; found: 583.0986.
Intermediate 182




N-(4-Fluoro-2-(3-hydroxyprop-1-ynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. Reaction of intermediate 174, N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.563 g, 1.00 mmol) with propargyl alcohol (0.18 ml, 3.2 mmol) using the conditions described for intermediate 178 gave 0.415 g (85% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.64 (6H, s, 2×CH3), 4.04 (4H, m, 2×CH2), 4.39 (2H, d, J=6.6 Hz, CH2), 4.68 (2H, d, J=6.0 Hz, CH2), 5.29 (2H, s, OCH2), 6.99 (1H, m, aromatic), 7.13 (1H, dd, J=2.5 Hz and J=9.1 Hz, aromatic), 7.22 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 7.33-7.37 (3H, m, aromatics), 7.44-7.47 (2H, m, aromatics), 7.68 (1H, broad t, NH). HRMS (ESI+) calculated for C27H27FN3O5 [M+H+]: 492.1935; found: 492.1939.
Intermediate 183




3-[2-((3-(Benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl]prop-2-ynyl methanesulfonate. A solution of intermediate 182, N-(4-fluoro-2-(3-hydroxyprop-1-ynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.280 g, 0.57 mmol) and triethylamine (0.12 ml, 0.86 mmol) in dichloromethane (5 ml) was cooled to 0° C., treated drop wise with methanesulfonyl chloride (0.050 ml, 0.64 mmol) and then stirred for 30 min. The reaction mixture was then diluted with ethyl acetate, washed with saturated sodium bicarbonate, brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in hexane) gave 0.245 g (75% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.65 (6H, s, 2×CH3), 3.10 (3H, s, SCH3), 4.05 (4H, m, 2×CH2), 4.67 (2H, d, J=6.1 Hz, NCH2), 5.03 (2H, s, CH2), 5.29 (2H, s, OCH2), 7.07 (1H, m, aromatic), 7.18 (1H, dd, J=2.5 Hz and J=9.1 Hz, aromatic), 7.32-7.36 (3H, m, aromatics), 7.38 (1H, dd, J=5.5 Hz and J=8.6 Hz, aromatic), 7.49-7.51 (2H, m, aromatics), 7.77 (1H, broad t, NH). MS (ESI+) m/e 570 [M+H+].
Intermediate 184




N-(2-(3-(Dimethylamino)prop-1-ynyl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 183, 3-[2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl]prop-2-ynyl methanesulfonate (0.100 g, 0.18 mmol) in acetonitrile (5 ml) was treated at 22° C. with 0.3 ml (0.6 mmol) of a 2 M solution of dimethylamine in tetrahydrofuran and the resulting mixture was stirred for 30 min. The reaction mixture was then diluted with ethyl acetate, washed with saturated sodium bicarbonate, brine, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give 0.080 g (88% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.64 (6H, s, 2×CH3), 2.43 (6H, s, 2×NCH3), 3.58 (2H, broad s, CH2), 4.05 (4H, m, 2×CH2), 4.70 (2H, d, J=6.0 Hz, NCH2), 5.30 (2H, s, OCH2), 7.01 (1H, m, aromatic), 7.18 (1H, dd, J=3 Hz and J=9.1 Hz, aromatic), 7.32-7.38 (4H, m, aromatics), 7.50-7.53 (2H, m, aromatics), 7.79 (1H, broad t, NH). HRMS (ESI+) calculated for C29H32FN4O4 [M+H+]: 519.2408; found: 519.2407.
Intermediate 185




N-(4-Fluoro-2-(3-(methylthio)prop-1-ynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 183, 3-[2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl]prop-2-ynyl methanesulfonate (0.160 g, 0.28 mmol) in N,N-dimethylformamide (3 ml) was treated at 0° C. with sodium thiomethoxide (0.026 g, 0.37 mmol) and the resulting mixture stirred for 2 h. The reaction mixture was then diluted with ethyl acetate, washed with saturated sodium bicarbonate and brine, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in hexane) gave 0.114 g (78% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.64 (6H, s, 2×CH3), 2.28 (3H, s, SCH3), 3.45 (2H, s, SCH2), 4.05 (4H, m, 2×CH2), 4.69 (2H, d, J=6.1 Hz, NCH2), 5.31 (2H, s, OCH2), 6.98 (1H, m, aromatic), 7.15 (1H, dd, J=3 Hz and J=9.1 Hz, aromatic), 7.32-7.38 (4H, m, aromatics), 7.50-7.53 (2H, m, aromatics), 7.80 (1H, broad t, NH). HRMS (ESI+) calculated for C28H29FN3O4S [M+H+]: 522.1863; found: 522.1844.
Intermediate 186




N-(4-Fluoro-2-(3-(methylsulfonyl)prop-1-ynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of intermediate 185, N-(4-fluoro-2-(3-(methylthio)prop-1-ynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.110 g, 0.21 mmol) in dichloromethane (3 ml) was treated at 0° C. with 3-chloroperoxybenzoic acid (0.120 g of 85%, 0.59 mmol) and the resulting mixture was stirred at 22° C. for 30 min. The reaction mixture was then diluted with ethyl acetate, washed with saturated sodium bicarbonate, brine, then dried over anhydrous magnesium sulfate and concentrated under reduced pressure. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in dichloromethane) gave 0.074 g (64% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.64 (6H, s, 2×CH3), 3.03 (3H, s, SCH3), 4.03 (2H, s, SCH2), 4.05 (4H, m, 2×CH2), 4.65 (2H, d, J=6.0 Hz, NCH2), 5.27 (2H, s, OCH2), 7.06 (1H, m, aromatic), 7.17 (1H, dd, J=3 Hz and J=8.6 Hz, aromatic), 7.31-7.38 (6H, m, aromatics), 8.12 (1H, broad t, NH). HRMS (ESI+) calculated for C28H29FN3O6S [M+H+]: 554.1761; found: 554.1784.
Intermediate 187




Diethyl 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate. A solution of intermediate 174, N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.200 g, 0.35 mmol) and triphenylphosphine (0.020 mg) in ethanol (5 ml) was flushed with argon and then treated with N,N-diisopropylethylamine (0.25 ml, 1.4 mmol), palladium(11) acetate (0.020 g) and diethyl phosphite (0.15 ml, 1.16 mmol). The reaction mixture was then sealed and heated at 80° C. for 18 h. The reaction mixture was then diluted with ethyl acetate, washed with 0.1 N hydrochloric acid, saturated sodium bicarbonate, brine, then dried over anhydrous magnesium sulfate and concentrated. Chromatography of the residue on silica gel (elution gradient of acetonitrile in dichloromethane) gave 0.103 g (51% yield) of the title compound as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.36 (6H, t, J=6.6 Hz, 2×CH3), 1.64 (6H, s, 2×CH3), 4.04 (4H, m, 2×CH2), 4.08-4.22 (4H, m, 2×OCH2), 4.78 (2H, d, J=6.6 Hz, NCH2), 5.29 (2H, s, OCH2), 7.21 (1H, m, aromatic), 7.29-7.34 (3H, m, aromatics), 7.45-7.52 (3H, m, aromatics), 7.65-7.72 (1H, m, aromatic), 8.67 (1H, broad t, NH). HRMS (ESI+) calculated for C28H34FN3O7P [M+H+]: 574.2118; found: 574.2126.
Intermediate 188




Ethyl hydrogen 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate. A solution of intermediate 187, diethyl 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate (0.115 g, 0.20 mmol) in tetrahydrofuran (3 ml)/ethanol (3 mL) was treated with 1 N aqueous sodium hydroxide (1.0 ml, 1.0 mmol) and the resulting mixture heated at 45° C. for 2 h. The reaction mixture was then diluted with ethyl acetate, washed with 0.1 N hydrochloric acid, brine, dried over anhydrous magnesium sulfate and concentrated. The residue was purified on a Shimadzu automated preparative HPLC system (column YMC Pack C-18, 5μ, 20×250 mm, elution gradient acetonitrile-water 0.1% trifluoroacetic acid) to give 0.070 g (64% yield) of the title material as a clear oil. 1HNMR 400 MHz (CDCl3) δ ppm: 1.37 (3H, t, J=7.1 Hz, CH3), 1.62 (6H, s, 2×CH3), 4.04 (4H, m, 2×CH2), 4.18 (2H, m, OCH2), 4.75 (2H, broad d, J=5 Hz, NCH2), 5.32 (2H, s, OCH2), 7.21 (1H, m, aromatic), 7.30-7.33 (3H, m, aromatics), 7.38-7.40 (2H, m, aromatics), 7.47-7.52 (1H, m, aromatic), 7.56-7.63 (1H, m, aromatic), 8.56 (1H, broad t, NH). HRMS (ESI+) calculated for C26H30FN3O7P [M+H+]: 546.1805; found: 546.1786.
Intermediate 189




N-(2-Acetamido-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of intermediate 169, N-(2-amino-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.033 g, 0.073 mmol) in dry tetrahydrofuran (10 mL) was added acetyl chloride (5.7 μL, 0.08 mmol) and diisopropylethylamine (38 μL, 0.22 mmol). The reaction mixture was stirred at 23° C. for 3 h. NaHCO3 (0.25 M, 20 mL) was then added and the organic material extracted with ethyl acetate (3×25 mL). The organic extracts were combined, dried (MgSO4) and concentrated in vacuo. The residue was purified on a Biotage system using a silica gel column with ethyl acetate:Hex (1:2 to 2:1) as eluent to afford the title compound (0.025 g, 69% yield). 1HNMR 400 MHz (MeOD) δ (ppm): 7.66 (1H, dd, J=10.5, 2.8 Hz), 7.43-7.25 (6H, m), 6.87 (1H, ddd, (dt), J=8.8, 2.6 Hz), 5.19 (2H, s), 4.47 (2H, m), 4.08 (2H, t, J=4.9 Hz), 3.99 (2H, t, J=4.9 Hz), 2.19 (3H, s), 1.62 (6H, s). LCMS (M+H)+ m/z 495.
Intermediate 190




2-((3-(Benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorobenzoic acid. To a suspension of intermediate 154, methyl 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorobenzoate. (500 mg, 1.01 mmol) in MeOH (10 mL) and CH3CN (5 mL) was added 1N NaOH (2 mL) and the mixture stirred at room temperature for 2 hrs. The mixture was concentrated in vacuo and the residue purified by reverse phase column chromatography (YMC, C-18 ODS, 10-25% CH3CN/H2O) to provide 90 mg (Yield 19%) of the title compound as an off-white powder: LC/MS m/z 482 (M+H).
Intermediate 191




N-(2-((2-Aminoethyl)carbamoyl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A mixture of intermediate 190, 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorobenzoic acid, (59 mg, 0.12 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate, HATU (76 mg, 0.2 mmol; Aldrich) in DMF (1 mL) was stirred for 20 min. To this mixture was added ethanolamine (20 mg, 0.3 mmol) and the stirring continued overnight. The mixture was concentrated in vacuo, dissolved in CH2Cl2, washed with water then dried (MgSO4), filtered, and concentrated to provide 55 mg (Yield 87%) of the title compound: LC/MS m/z 525 (M+H).
Intermediate 192




(R)—N-(4-Fluoro-2-(2-(hydroxymethyl)-5-oxopyrrolidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a stirred solution of intermediate 166, (R)—N-(2-(2-((tert-butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.10 g, 0.150 mmol) in tetrahydrofuran (5 mL) at 23° C. was added a solution of tetrabutylammonium fluoride (1M in tetrahydrofuran) (180 μL, 0.18 mmol). The reaction mixture was stirred at 23° C. for 3 h. NaHCO3 (1N in H2O, 30 mL) was then added and the organic material was extracted with ethyl acetate (2×25 mL). The combined organic extracts were dried (MgSO4), filtered and concentrated in vacuo. The residue was purified on a Biotage system using a silica gel column with ethyl acetate/Hexanes (1:1) to ethyl acetate 100% as eluent to afford the title compound (0.060 g, 73% yield): 1HNMR 400 MHz (MeOD) 8 (ppm): 7.55 (1H, dd, (t), 6.6 Hz), 7.43 (2H, m), 7.13 (1H, dd, J=9.5, 2.4 Hz), 7.03 (1H, m), 5.26 (2H, s), 4.57 (1H, m), 4.32 (2H, m), 4.08 (2H, t, J=5.0 Hz), 7.03 (2H, t, J=5.0 Hz), 3.53 (2H, m), 2.69-2.56 (2H, m), 2.38 (1H, m), 2.22 (1H, m), 1.62 (6H, s). LCMS (M+H)+ m/z 551.
Intermediate 193




(R)-(1-(2-((3-(Benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl)-5-oxopyrrolidin-2-yl)methyl acetate. To a solution of intermediate 192, (R)—N-(4-fluoro-2-(2-(hydroxymethyl)-5-oxopyrrolidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.045 g, 0.082 mmol) in tetrahydrofuran (5 mL) was added acetyl chloride (12.8 μL, 0.180 mmol) and diisopropylethylamine (31.4 μL, 0.180 mmol). The reaction mixture was stirred at 23° C. for 4 h. NaHCO3 (1N in H2O, 30 mL) was then added and the organic material was extracted with ethyl acetate (2×25 mL). The combined organic extracts were dried (MgSO4), filtered and concentrated in vacuo. The residue was purified on a Biotage system using a silica gel column with ethyl acetate/Hexanes (1:1) to ethyl acetate 100% as eluent to afford the title compound (0.032 g, 67% yield): 1HNMR 400 MHz (MeOD) δ ppm: 7.48 (3H, m), 7.33 (3H, m), 7.16 (1H, m), 7.03 (1H, m), 5.24 (2H, s), 4.60 (1H, m), 4.55 (1H, m), 4.22 (1H, dd, J=12.0, 4.5 Hz), 4.08 (2H, t, J=5.0 Hz), 3.99 (3H, m), 2.62 (2H, m), 2.44 (1H, m), 2.09-1.92 (5H, m), 1.62 (3H, s), 1.61 (3H, s). LCMS (M+H)+ m/z 593.
Intermediate 194




(R)-(1-(2-((3-(Benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl)-5-oxopyrrolidin-2-yl)methyl methanesulfonate. To a stirred solution of intermediate 192, (R)—N-(4-fluoro-2-(2-(hydroxymethyl)-5-oxopyrrolidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.160 g, 0.291 mmol) in CH2Cl2 (10 mL) at 0° C. was added triethylamine (81 μL, 0.582 mmol) and methanesulfonyl chloride (27 μL, 0.349 mmol). The reaction mixture was stirred at 23° C. for 4 h. Water (50 mL) was then added and the organic material was extracted with CH2Cl2 (2×50mL). The combined organic extracts were dried (MgSO4), filtered and concentrated in vacuo to afford the title compound (0.178 g, 98% yield). The crude material was used without further purification for the next step: LCMS (M+H)+ m/z 629.
Intermediate 195




(R)—N-(2-(2-(Azidomethyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of intermediate 194, (R)-(1-(2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl)-5-oxopyrrolidin-2-yl)methyl methanesulfonate (0.150 g, 0.239 mmol) in DMF (10 mL) was added sodium azide (0.019 g, 0.287 mmol). The resulting mixture was stirred at 50° C. for 6 h. Water (50 mL) was then added and the organic material was extracted with ethyl acetate (2×50 mL). The combined organic extracts were washed with H2O (50 mL), dried (MgSO4), filtered and concentrated in vacuo to afford the title compound (0.125 g, 91% yield). LCMS (M+H)+ m/z 576.
Intermediate 196




N-(Benzo[b]thiophen-1,1-dione-7-ylmethyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of intermediate 147, N-(benzo[b]thiophen-7-ylmethyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.100 g, 0.21 mmol) in dichloroethane (10 mL) was added per-acetic acid (32% in H2O) (1.0 mmol, 200 μL). The reaction mixture was stirred at 23° C. for 48 h. Water (50 mL) was added and the organic material was extracted with CH2Cl2 (2×50 mL). The combined organic extracts were dried (MgSO4), filtered and concentrated in vacuo to afford the title compound (0.106 g, 99% yield): 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.11 (1H, t, J=6.4 Hz), 7.65 (1H, d, J=6.8 Hz) 7.59-7.29 (9H, m), 5.15 (2H, s), 4.77 (2H, d, J=6.5 Hz), 4.04 (2H, t, J=5.0 Hz), 3.90 (2H, m), 1.59 (6H, s). LCMS (M+H)+ m/z 508.
Intermediate 197




1-(Azidomethyl)-4-fluoro-2-iodobenzene. A solution of 1-(bromomethyl)-4-fluoro-2-iodobenzene (M. Protiva et al., Collect. Czech. Chem. Comm., 44, 1979, 2108-2123) (17.9 g, 56.8 mmol) in N,N-dimethylformamide (35 ml) was treated with sodium azide (5.0 g, 76.7 mmol) and the resulting mixture was heated to 50° C. for 4 h. The cooled mixture was filtered, the filtrate was concentrated in vacuo and the residue was chromatographed on silica gel (elution hexane) to give 15.7 g (97% yield) of the title azide as a clear oil. 1HNMR 400 MHz (DMSO-d6) δ (ppm): 4.53 (2H, s, CH2), 7.32 (1H, m, aromatic), 7.54 (1H, dd, J=6 Hz and J=8.6 Hz, aromatic), 7.83 (1H, dd, J=3 Hz and J=8 Hz, aromatic).
Intermediate 198




(4-Fluoro-2-iodophenyl)methanamine. A solution of 1-(azidomethyl)-4-fluoro-2-iodobenzene (15.2 g, 54.8 mmol) in DMF (35 ml) at 0° C. was treated with triphenylphosphine (21.6 g, 81.2 mmol) and then stirred for 1 h. The reaction mixture was then treated with water (5 ml) and heated at 55° C. for 1 h. The DMF was concentrated in vacuo and the residue was diluted with ethyl acetate (200 ml). The organic phase was extracted with 0.5 N hydrochloric acid (140 ml) and the aqueous extract was washed with ethyl acetate. The aqueous phase was then adjusted to pH 9 with 1 N LiOH and extracted with ethyl acetate (2×200 ml). The combined organic phases were dried over anhydrous magnesium sulfate and concentrated. The residue was diluted with ether (200 ml), filtered and concentrated. Distillation of the residue in vacuo gave 8.52 g (62% yield) of the title amine as a clear oil: bp 85° C./0.35 torr (bulb to bulb distillation air bath temperature). 1HNMR 400 MHz (DMSO-d6) δ (ppm): 3.64 (2H, s, CH2), 7.27 (1H, m, aromatic), 7.53 (1H, dd, J=6 Hz and J=8.6H, aromatic), 7.83 (1H, dd, J=3 Hz and J=8 Hz, aromatic).
Intermediate 199




tert-Butyl 4-fluoro-2-iodobenzylcarbamate. A solution of (4-fluoro-2-iodophenyl)methanamine (21.4 g, 85.2 mmol) in dichloromethane (350 ml) was treated at 0° C. with di-tert-butyl dicarbonate (20.5 g, 93.8 mmol) followed by triethylamine added drop wise over 30 min. The resulting mixture was then allowed to warm up to 25° C. and stirred for 18 h. The reaction mixture was then washed with water, brine, dried over anhydrous magnesium and concentrated. Chromatography of the residue on silica gel (elution gradient of ethyl acetate 5-20% in hexane) gave 28.37 g (95% yield) of the title carbamate as a clear oil. 1HNMR 400 MHz (CDCl3) (ppm): 1.47 (9H, s, t-Bu), 4.32 (2H, d, J=6.0 Hz, NCH2), 5.04 (1H, broad, NH), 7.07 (1H, m, aromatic), 7.35 (1H, m, aromatic), 7.56 (1H, dd, J=2.8 Hz and J=8 Hz, aromatic).
Intermediate 200




tert-Butyl 2-(dimethoxyphosphoryl)-4-fluorobenzylcarbamate. A solution of tert-butyl 4-fluoro-2-iodobenzylcarbamate (5.00 g, 14.24 mmol), dimethyl phosphite (4.70 g, 42.7 mmol) and N,N-diisopropylethylamine (9.9 ml, 56.8 mmol) in methanol (75 ml) was flushed with argon and then treated with triphenylphosphine (0.5 g) and palladium(II) acetate (0.75 g). The resulting mixture was then sealed and heated at 100° C. for 1 hour. The cooled reaction mixture was concentrated in vacuo, diluted with ethyl acetate, washed with water, brine, dried over anhydrous magnesium and concentrated. Chromatography of the residue on silica gel (elution gradient of acetonitrile in dichloromethane) gave 3.24 g (68% yield) of the title phosphonate as a clear oil. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.44 (9H, s, t-Bu), 3.81 (3H, s, OCH3), 3.84 (3H, s, OCH3), 4.49 (2H, d, J=6.0 Hz, NCH2), 5.7 (1H, broad, NH), 7.24 (1H, m, aromatic), 7.47-7.7 (2H, m, aromatics). HRMS (ESI+) calculated for C14H22FNO5P [M+H+]: 334.1220; found: 334.1217.
Intermediate 201




Dimethyl 2-(aminomethyl)-5-fluorophenylphosphonate trifluoroacetic acid salt. A solution of tert-butyl 2-(dimethoxyphosphoryl)-4-fluorobenzylcarbamate (0.140 g, 0.42 mmol) in dichloromethane (5 ml) was treated with trifluoroacetic acid (5 ml) and the resulting mixture was stirred at 25° C. for 1 h. The solvent was then evaporated in vacuo to give the title amine salt as an amorphous white solid. MS (ESI+) m/z 234 [M+H+].
Intermediate 202




Benzyl hydrogen 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate. A solution of N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.200 g, 0.35 mmol) in dimethyl sulfoxide (5 ml) was flushed with argon and then treated triethylamine (0.25 ml, 1.8 mmol), tetrakis(triphenylphosphine)palladium(0) (0.10 g) and dibenzyl phosphite (0.25 ml, 1.06 mmol). The reaction mixture was then sealed and heated at 80° C. for 16 h. The reaction mixture was then diluted with ethyl acetate, washed with 0.1 N hydrochloric acid, brine, dried over anhydrous magnesium sulfate and concentrated. Purification of the residue by preparative HPLC (column YMC Pack C-18, 5μ, 20×250 mm, elution gradient acetonitrile-water 0.1% trifluoroacetic acid) gave 0.150 g (60% yield) of the title material as a white solid; mp 142° C. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.58 (6H, s, 2×CH3), 4.0 (4H, m, 2×CH2), 4.73 (2H, broad s, OCH2), 5.05 (2H, d, J=8.6 Hz, NCH2), 5.28 (2H, s, OCH2), 7.20 (1H, m, aromatic), 7.29-7.34 (6H, m, aromatics), 7.45-7.64 (6H, m, aromatics), 8.24 (1H, broad, NH). MS (ESI+) m/z 608 [M+H+].
Intermediate 203




N-(4-Fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. White solid (77% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.01 (3H, t, J=7.4 Hz, CH3), 1.9-2.0 (1H, m, CH), 2.2-2.35 (1H, m, CH), 2.47 (3H, s, CH3), 3.8-3.9 (2H, m, CH2), 4.1-4.35 (2H, m, CH2), 4.4-4.5 (2H, m, NCH2), 4.55 (1H, m, CH), 5.32 (2H, s, OCH2), 7.07 (1H, dd, J=2.6 Hz and J=8.6 Hz, aromatic), 7.17 (11H, m, aromatic), 7.24-7.28 (3H, m, aromatics), 7.4-7.45 (2H, m, aromatics), 7.71 (1H, dd, J=6.0 Hz and J=8.6 Hz, aromatic), 8.31 (1H, s, CH), 8.48 (1H, broad t, NH). HRMS (ESI+) calculated for C27H28FN6O4 [M+H+]: 519.2156; found: 519.2136.
Intermediate 204




N-(4-Fluoro-2-(N,N-dimethylsulfamoyl)benzyl)-3-(benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. White solid (95% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.0 (3H, t, J=7.4 Hz, CH3), 1.9-2.05 (1H, m, CH), 2.2-2.3 (1H, m, CH), 2.88 (6H, s, 2×NCH3), 3.8-3.9 (2H, m, CH2), 4.1-4.35 (2H, m, CH2), 4.52 (1H, m, CH), 4.82 (2H, d, J=6.6 Hz, NCH2), 5.36 (2H, s, OCH2), 7.24 (1H, m, aromatic), 7.28-7.31 (3H, m, aromatics), 7.46-7.49 (2H, m, aromatics), 7.54 (1H, dd, J=2.6 Hz and J=8.6 Hz, aromatic), 7.74 (1H, dd, J=5.4 Hz and J=8.6 Hz, aromatic), 8.39 (1H, broad t, NH). HRMS (ESI+) calculated for C26H30FN4O6S [M+H+]: 545.1870; found: 545.1859.
Intermediate 205




N-(4-Fluoro-2-(5-methyl-1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. White crystals; mp 163° C. (ethyl acetate-hexane). (70% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.03 (3H, t, J=7.4 Hz, CH3), 1.9-2.05 (1H, m, CH), 2.25-2.35 (1H, m, CH), 2.46 (3H, s, CH3), 3.75-3.9 (2H, m, CH2), 4.1-4.35 (4H, m, CH2 and N CH2), 4.53 (1H, m, CH), 5.35 (2H, ABq, JAB=10.9 Hz), 7.0 (1H, dd, J=2.6 Hz and J=8.6 Hz, aromatic), 7.20 (1H, m, aromatic), 7.28-7.32 (3H, m, aromatics), 7.45-7.5 (2H, m, aromatics), 7.69 (1H, dd, J=6.0 Hz and J=8.6 Hz, aromatic), 7.89 (1H, s, CH), 8.29 (1H, broad t, NH). Anal. Calcd for C27H27FN6O4: C, 62.54; H, 5.24; N, 16.20; Found: C, 62.39; H, 5.36; N, 16.19.
Intermediate 206




N-(4-Fluoro-2-(1-methyl-1H-pyrazol-3-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. A solution of N-(4-fluoro-2-(1H-pyrazol-5-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.185 g, 0.36 mmol) in N,N-dimethylformamide (3 ml) was treated with iodomethane (0.025 ml, 0.4 mmol) and potassium carbonate (0.076 g, 0.55 mmol) and the resulting mixture was stirred at 25° C. for 5 h. The reaction mixture was then diluted with ethyl acetate, washed with water, brine, dried over anhydrous magnesium and concentrated. Chromatography of the residue on silica gel (elution gradient of ethyl acetate in dichloromethane) gave 0.120 g (64% yield) of the title compound as a white solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.62 (6H, s, 2×CH3), 3.84 (3H, s, NCH3), 3.95-4.05 (4H, m, 2×CH2), 4.69 (2H, d, J=6.1 Hz, NCH2), 5.21 (2H, s, OCH2), 6.49 (1H, d, J=2.2 Hz, CH), 7.01 (1H, m, aromatic), 7.24-7.3 (4H, m, aromatics), 7.40 (1H, d, J=2.2 Hz, CH), 7.42-7.45 (2H, m, aromatics), 7.64 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 8.75 (1H, broad t, NH). HRMS (ESI+) calculated for C28H29FN5O4 [M+H+]: 518.2204: found: 518.2222.
Intermediate 207




N-(4-Fluoro-2-(1-methyl-1H-pyrazol-5-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. N-(4-Fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.250 g, 0.44 mmol) was reacted with 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.183 g, 0.88 mmol), sodium carbonate (0.150 g, 1.41 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.10 g) to give 0.152 g (65% yield) of the title material as a white solid after chromatography on silica gel. 1H NMR 400 MHz (CDCl3) δ (ppm): 1.63 (6H, s, 2×CH3), 3.69 (3H, s, NCH3), 4.0-4.1 (4H, m, 2×CH2), 4.34 (2H, d, J=6.6 Hz, NCH2), 5.29 (2H, s, OCH2), 6.26 (1H, d, J=2.0 Hz, CH), 6.98 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.13 (1H, m, aromatic), 7.3-7.35 (3H, m, aromatics), 7.47-7.55 (4H, m, aromatics and NH), 7.57 (1H, d, J=2.0 Hz, CH). MS (ESI+) m/z 518 [M+H+].
Intermediate 208




N-(2-(3-Methyl-1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. White crystals; mp 213-214° C. (ethyl acetate). (90% yield). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.65 (6H, s, 2×CH3), 2.50 (3H, s, CH3), 3.99-4.15 (4H, m, 2×CH2), 4.50 (2H, d, J=5=6.5 Hz, NCH2), 5.31 (2H, s, OCH2), 7.26-7.34 (4H, m, aromatics), 7.4-7.52 (4H, m, aromatics), 7.73-7.77 (1H, m, aromatic), 8.27 (1H, s, CH), 8.46 (1H, broad t, NH). Anal. Calcd for C27H28N6O4: C, 64.78; H, 5.63; N, 16.79; Found: C, 64.67; H, 5.41; N, 16.55.
Intermediate 209




Methyl hydrogen 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate. A solution of N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.200 g, 0.35 mmol) in acetonitrile (5 ml) was flushed with argon and then treated with N,N-diisopropylethylamine (0.25 ml, 1.4 mmol), triphenylphosphine (0.030 g), palladium(II) acetate (0.040 g). and dimethyl phosphite (0.10 ml, 1.06 mmol). The reaction mixture was then heated in a microwave oven at 120° C. for 40 min. The reaction mixture was then diluted with ethyl acetate, washed with 0.1 N hydrochloric acid, brine, dried over anhydrous magnesium sulfate and concentrated. Purification of the residue by preparative HPLC (column YMC Pack C-18, 5μ, 20×250 mm, elution gradient acetonitrile-water 0.1% trifluoroacetic acid) gave 0.120 g (64% yield) of the title material as a white solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.62 (6H, s, 2×CH3), 3.77 and 3.79 (3H, 2 s, OCH3), 3.95-4.05 (4H, m, 2×CH2), 4.75 (2H, d, J=4.6 Hz, NCH2), 5.32 (2H, s, OCH2), 7.22 (1H, m, aromatic), 7.3-7.35 (3H, m, aromatics), 7.38-7.42 (2H, m, aromatics), 7.48-7.54 (1H, m, aromatic), 7.55-7.62 (1H, m, aromatic), 8.54 (1H, broad t, NH). HRMS (ESI+) calculated for C25H28FN3O7P [M+H+]: 532.1649; found: 532.1642.
Intermediate 210 and 211




A solution of N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.200 g, 0.35 mmol), 1,3,2-dioxaphosphorinan-2-one (0.115 g, 1.06 mmol) and N,N-diisopropylethylamine (0.25 ml, 1.4 mmol) in acetonitrile (5 ml) was flushed with argon and then treated with triphenylphosphine (0.025 g) and palladium(II) acetate (0.040 g). The resulting mixture was then sealed and heated at 120° C. in a microwave oven for 20 min. The cooled reaction mixture was concentrated and chromatographed on silica gel (elution gradient of acetonitrile in dichloromethane) to give 0.073 g (38% yield) of intermediate 210 as an amorphous solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.65 (6H, s, 2×CH3), 3.97-4.05 (4H, m, 2×CH2), 4.34-4.42 (2H, m, CH2), 4.62-4.67 (2H, m, CH2), 4.80 (2H, d, J=6.5 Hz, NCH2), 5.30 (2H, s, OCH2), 7.22-7.4 (5H, m, aromatics), 7.52-7.57 (2H, m, aromatics), 7.71-7.78 (1H, m, aromatic), 8.77 (1H, broad t, NH). MS (ESI+) m/z 544 [M+H+].
In an alternative work-up, the above crude reaction mixture was diluted with ethyl acetate, washed with 0.1 N hydrochloric acid, brine, dried over anhydrous magnesium sulfate and concentrated. Purification of the residue by preparative HPLC (column YMC Pack C-18, 5μ, 20×250 mm, elution gradient acetonitrile-water 0.1% trifluoroacetic acid) gave 0.092 g (45% yield) of 2-hydroxyethyl hydrogen 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate, intermediate 211 as a white solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.61 (6H, s, 2×CH3), 3.77 (2H, broad m, CH2), 3.95-4.05 (4H, m, 2×CH2), 4.10 (2H, broad m, CH2), 4.81 (2H, broad, NCH2), 5.25 (2H, s, OCH2), 7.1-7.19 (1H, m, aromatic), 7.29-7.32 (3H, m, aromatics), 7.4-7.47 (3H, m, aromatics), 7.55-7.64 (1H, m, aromatic), 8.27 (1H, broad t, NH). HRMS (ESI+) calculated for C26H30FN3O8P [M+H+]: 562.1755; found: 562.1774.
Intermediate 212




Bis(2-Hydroxyethyl) 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate. A solution of N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.200 g, 0.35 mmol), 1,3,2-dioxaphosphorinan-2-one (0.115 g, 1.06 mmol) and N,N-diisopropylethylamine (0.25 ml, 1.4 mmol) in ethylene glycol (5 ml) was flushed with argon and then treated with triphenylphosphine (0.025 g) and palladium(11) acetate (0.040 g). The resulting mixture was then sealed and heated at 80° C. for 2 hour. The cooled reaction mixture was diluted with ethyl acetate, washed with water, brine, dried over anhydrous magnesium and concentrated. The residue was purified by preparative HPLC (column YMC Pack C-18, 5μ, 20×250 mm, elution gradient acetonitrile-water 0.1% trifluoroacetic acid) to give 0.093 g (44% yield) of the title phosphonate as a clear oil. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.63 (6H, s, 2×CH3), 3.8-3.9 (4H, m, 2×CH2), 4.0-4.1 (4H, m, 2×CH2), 4.15-4.3 (4H, m, 2×CH2), 4.81 (2H, d, J=6.1 Hz, NCH2), 5.29 (2H, s, OCH2), 7.22-7.4 (6H, m, aromatics), 7.5-7.6 (2H, m, aromatics), 8.55 (1H, broad t, NH). HRMS (ESI+) calculated for C28H34FN3O9P [M+H+]: 606.2017; found: 606.2018.
Intermediate 213




Reaction of N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.350 g, 0.62 mmol) and 5,5-dimethyl-1,3,2-dioxaphosphorinan-2-one (0.280 g, 1.86 mmol) with triphenylphosphine (0.035 g) and palladium(II) acetate (0.050 g) at 80° C. as described above gave 0.140 g (39% yield) of intermediate 213 as a white solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.13 (3H, s, CH3), 1.16 (3H, s, CH3), 1.61 (6H, s, 2×CH3), 3.9-4.05 (6H, m, 3×CH2), 4.36 (2H, t, J=10.5 Hz, CH2), 4.83 (2H, d, J=6.6 Hz, NCH2), 5.36 (2H, s, OCH2), 7.25-7.33 (4H, m, aromatics), 7.4-7.5 (3H, m, aromatics), 7.7-7.75 (1H, m, aromatic), 8.76 (1H, broad t, NH). HRMS (ESI+) calculated for C29H34FN3O7P [M+H+]: 586.2118; found: 586.2133.
Intermediate 214




2-Azido-1-bromo-4-fluorobenzene. 2-Bromo-5-fluoro aniline (2.00 g, 10.53 mmol) was dissolved in concentrated HCl (10 mL) and water (10 mL) and cooled to 0° C. Aqueous NaNO2 solution (1.090 g, 15.8 mmol of NaNO2 in 10 mL of water) was added dropwise at such a rate that the temperature did not exceed 5° C. This mixture was stirred at 0° C. for 1.5 h. A solution of NaN3 (1.027 g, 15.8 mmol) and NaOAc (12.95 g, 158 mmol) in water (50 mL) was then added at 0-5° C. and the mixture was stirred for an additional 1 h at this temperature. The mixture was extracted with ethyl acetate and the combined extracts were washed with brine and dried over Na2SO4. The filtrate was concentrated to afford the title compound as a tan solid (2.188 g, 96%): 1H NMR (400 MHz, CDCl3): 7.53 (1H, dd, J=8.8, 5.6 Hz), 6.94 (1H, dd, J=8.8, 2.8 Hz), 6.79 (1H, ddd, J=8.8, 7.6, 2.8 Hz).
Intermediate 215




1-(2-Bromo-5-fluorophenyl)-4-(trimethylsilyl)-1H-1,2,3-triazole. A mixture of 2-azido-1-bromo-4-fluorobenzene (1.047 g, 4.85 mmol) and trimethylsilylacetylene (2.01 mL, 14.54 mmol) in toluene (5 mL) was heated in a pressure vessel at 110° C. for 21.5 h. The reaction mixture was concentrated in vacuo and the residue was purified with a Biotage system on silica gel with hexanes:ethyl acetate (9:1) gradient as the eluent to afford the title compound as a colorless oil (1.451 g, 95% yield): 1H NMR (400 MHz, CDCl3) δ ppm: 7.97 (1H, s), 7.74 (1H, dd, J=9.0, 5.4 Hz), 7.37 (1H, dd, J=8.5, 2.9 Hz), 7.16 (1H, ddd, J=8.8, 7.6, 3.0 Hz), 0.40 (9H, s), LCMS (+ESI, M+H+) m/z 314/316.
Intermediate 216




1-(2-Bromo-5-fluorophenyl)-5-methyl-4-(trimethylsilyl)-1H-1,2,3-triazole. The title compound can be prepared according to the procedure provided for 1-(2-bromo-5-fluorophenyl)-4-(trimethylsilyl)-1H-1,2,3-triazole 1H NMR (400 MHz, CDCl3) δ ppm: 7.73-7.69 (1H, m), 7.20-7.16 (2H, m), 2.22 (3H, s), 0.39 (9H, s); LCMS (+ESI, M+H+) m/z 328/330.
Intermediate 217




1-(2-Bromo-5-fluorophenyl)-4-tert-butyl-1H-1,2,3-triazole. The title compound can be prepared according to the procedure provided for 1-(2-bromo-5-fluorophenyl)-4-(trimethylsilyl)-1H-1,2,3-triazole 1H NMR (400 MHz, CDCl3) δ ppm: 7.68-7.75 (2H, m), 7.38 (1H, dd, J=8.5, 2.9 Hz), 7.14 (1H, ddd, J=8.8, 7.6, 3.0 Hz), 1.43 (9H, s), LCMS (+ESI, M+H+) m/z 298/300.
Intermediate 218




1-(2-Bromo-5-fluorophenyl)-4-((tert-butyldimethylsilyloxy)methyl)-1H-1,2,3-triazole. The title compound can be prepared according to the procedure provided for 1-(2-bromo-5-fluorophenyl)-4-(trimethylsilyl)-1H-1,2,3-triazole 1H NMR (400 MHz, CDCl3) δ ppm: 7.95 (1H, s), 7.75 (1H, dd, J=9.0, 5.4 Hz), 7.38 (1H, dd, J=8.3, 3.0 Hz), 7.17 (1H, ddd, J=9.0, 7.5, 2.8 Hz), 4.96 (2H, s), 0.93 (9H, s), 0.13 (6H, s), LCMS (+ESI, M+H+) m/z 386/388.
Intermediate 219




1-(2-Bromo-5-fluorophenyl)-5-((tert-butyldimethylsilyloxy)methyl)-1H-1,2,3-triazole. The title compound can be prepared according to the procedure provided for 1-(2-bromo-5-fluorophenyl)-4-(trimethylsilyl)-1H-1,2,3-triazole 1H NMR (400 MHz, CDCl3) δ ppm: 7.70-7.78 (2H, m), 7.19-7.30 (2H, m), 4.67 (2H, s), 0.82 (9H, s), −0.03 (6H, s), LCMS (+ESI, M+H+) m/z 386/388.
Intermediate 220




1-(2-Bromo-5-fluorophenyl)-1H-1,2,3-triazole. 1-(2-Bromo-5-fluorophenyl)-4-(trimethylsilyl)-1H-1,2,3-triazole (0.800 g, 2.55 mmol) was dissolved in THF (10 mL) and tetrabutylammonium fluoride (2.8 mL, 2.80 mmol, 1.0 M in THF) was added dropwise and the reaction mixture was stirred at 25° C. for 4 h. The resulting mixture was concentrated in vacuo and the residue was purified with a Biotage system on silica gel with hexanes:ethyl acetate (8:2 to 7:3) gradient as the eluent to afford the title compound as a white solid (0.36 g, 58% yield): 1H NMR (400 MHz, CDCl3) δ ppm: 8.06 (1H, d, J=1.0 Hz), 7.90 (1H, d, J=1.3 Hz), 7.76 (1H, dd, J=8.8, 5.3 Hz), 7.39 (1H, dd, J=8.3, 2.8 Hz), 7.19 (1H, ddd, J=8.9, 7.5, 3.0 Hz), LCMS (+ESI, M+H+) m/z 242/244.
Intermediate 221




1-(2-Bromo-5-fluorophenyl)-5-methyl-1H-1,2,3-triazole. The title compound can be prepared according to the procedure provided for 1-(2-bromo-5-fluorophenyl)-1H-1,2,3-triazole 1H NMR (400 MHz, CDCl3) δ ppm: 7.76 (1H, dd, J=9.1, 5.3 Hz), 7.62 (1H, s), 7.19-7.26 (2H, m), 2.25 (3H, s), LCMS (+ESI, M+H+) m/z 256/258.
Intermediate 222




(1-(2-Bromo-5-fluorophenyl)-1H-1,2,3-triazol-4-yl)methanol. The title compound can be prepared according to the procedure provided for 1-(2-bromo-5-fluorophenyl)-1H-1,2,3-triazole 1H NMR (400 MHz, CDCl3) δ ppm: 8.02 (1H, s), 7.75 (1H, dd, J=8.8, 5.3 Hz), 7.38 (1H, dd, J=8.2, 2.9 Hz), 7.19 (1H, ddd, J=8.9, 7.5, 3.0 Hz), 4.94 (2H, s), LCMS (+ESI, M+H+) m/z 270/272.
Intermediate 223




4-Fluoro-2-(1H-1,2,3-triazol-1-yl)benzonitrile. A mixture of 1-(2-bromo-5-fluorophenyl)-1H-1,2,3-triazole (0.603 g, 2.49 mmol), CuCN (0.245 g, 2.74 mmol), and 15 mL of NMP was subjected to microwave irradiation at 150° C. for 0.5 h. The brown mixture was filtered over celite and washed with DMF. This solution was treated with 10% aqueous NH4OH (28-30% solution) and extracted with EtOAc. The combined organic phase were successively washed with 10% aqueous NH4OH (28-30% solution), saturated aqueous NH4Cl, water, brine and dried over Na2SO4. The resulting mixture was concentrated in vacuo and the residue was purified with a Biotage system on silica gel with hexanes:ethyl acetate (7:3 to 6:4) gradient as the eluent to afford the title compound as a light yellow solid (0.285 g, 61% yield): 1H NMR (400 MHz, CDCl3) δ ppm: 8.40 (1H, d, J=1.0 Hz), 7.96 (1H, s), 7.91 (1H, dd, J=8.6, 5.6 Hz), 7.77 (1H, dd, J=8.7, 2.4 Hz), 7.31-7.39 (1H, m).
Intermediate 224




4-Fluoro-2-(5-methyl-1H-1,2,3-triazol-1-yl)benzonitrile. The title compound can be prepared according to the procedure provided for 4-fluoro-2-(1H-1,2,3-triazol-1-yl)benzonitrile 1H NMR (400 MHz, CDCl3) δ ppm: 7.93 (1H, dd, J=8.7, 5.4 Hz), 7.67 (1H, s), 7.44 (1H, ddd, J=8.6, 7.6, 2.5 Hz), 7.35 (1H, dd, J=8.1, 2.5 Hz), 2.39 (3H, s), LCMS (+ESI, M+H+) m/z 203.
Intermediate 225




2-(4-tert-Butyl-1H-1,2,3-triazol-1-yl)-4-fluorobenzonitrile. The title compound can be prepared according to the procedure provided for 4-fluoro-2-(1H-1,2,3-triazol-1-yl)benzonitrile 1H NMR (400 MHz, CDCl3) δ ppm: 8.07 (1H, s), 7.87 (1H, dd, J=8.8, 5.6 Hz), 7.74 (1H, dd, J=8.8, 2.5 Hz), 7.25-7.33 (1H, m), 1.45 (9H, s), LCMS (+ESI, M+H+) m/z 245.
Intermediate 226




4-Fluoro-2-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)benzonitrile. The title compound can be prepared according to the procedure provided for 4-fluoro-2-(1H-1,2,3-triazol-1-yl)benzonitrile H NMR (400 MHz, CDCl3) δ ppm: 8.34 (1H, s), 7.91 (1H, dd, J=8.7, 5.4 Hz), 7.74 (1H, dd, J=8.6, 2.5 Hz), 7.35 (1H, ddd, J=8.8, 7.3, 2.5 Hz), 4.96 (2H, s), LCMS (+ESI, M+H+) m/z 219.
Intermediate 227




(4-Fluoro-2-(1H-1,2,3-triazol-1-yl)phenyl)methanamine hydrochloride. 1H NMR (400 MHz, DMSO-D6) δ ppm: 8.73 (1H, d, J=1.0 Hz), 8.53 (3H, brs), 8.07 (1H, d, J=1.0 Hz), 7.91 (1H, dd, J=8.7, 5.9 Hz), 7.66 (1H, dd, J=9.2, 2.7 Hz), 7.60 (1H, td, J=8.5, 2.7 Hz), 3.92 (2H, q, J=5.6 Hz), LCMS (+ESI, M+H+) m/z 193.
Intermediate 228




(4-Fluoro-2-(5-methyl-1H-1,2,3-triazol-1-yl)phenyl)methanamine hydrochloride. 1H NMR (400 MHz, DMSO-D6) δ ppm: 8.61 (2H, s), 7.98 (1H, dd, J=9.5, 6.2 Hz), 7.80 (1H, s), 7.61-7.67 (2H, m), 3.66 (2H, q, J=5.7 Hz), 2.26 (3H, s), LCMS (+ESI, M+H+) m/z 206.
Intermediate 229




(2-(4-tert-Butyl-1H-1,2,3-triazol-1-yl)-4-fluorophenyl)methanamine hydrochloride. 1H NMR (400 MHz, DMSO-D6) δ ppm: 8.55 (2H, brs), 8.52 (1H, s), 7.89 (1H, dd, J=8.8, 6.1 Hz), 7.65 (1H, dd, J=9.3, 2.5 Hz), 7.55 (1H, td, J=8.5, 2.5 Hz), 3.95 (2H, q, J=5.6 Hz), 1.36 (9H, s), LCMS (+ESI, M+H+) m/z 249.
Intermediate 230




(1-(2-(aminomethyl)-5-fluorophenyl)-1H-1,2,3-triazol-4-yl)methanol hydrochloride. 1H NMR (400 MHz, DMSO-D6) δ ppm: 8.56 (1H, s), 8.50 (2H, s), 7.89 (1H, dd, J=8.7, 5.9 Hz), 7.63 (1H, dd, J=9.2, 2.7 Hz), 7.58 (1H, td, J=8.5, 2.8 Hz), 4.65 (2H, s), 3.94 (2H, q, J=5.4 Hz), LCMS (+ESI, M+H+) m/z 223.
Intermediate 231




N-(2-Cyano-5-fluorophenyl)ethanesulfonamide. 1H NMR 400 MHz (CDCl3) (ppm): 7.65 (1H, dd, J=8.6, 5.8 Hz), 7.55 (1H, dd, J=10.2, 2.4 Hz), 6.96 (1H, ddd, J=8.7, 7.5, 2.5 Hz), 3.27 (3H, q, J=7.3 Hz), 1.47 (4H, t, J=7.3 Hz).
Intermediate 232




N-(2-(aminomethyl)-5-fluorophenyl)ethanesulfonamide hydrochloride. 1H NMR 400 MHz (CDCl3) δ (ppm): 7.56-7.64 (1H, m), 7.11-7.21 (2H, m) 4.25 (2H, s), 3.15 (2H, q, J=−7.3 Hz), 1.36 (3H, t, J=7.5 Hz).
Intermediate 233




(2-Bromo-5-fluoro-phenyl)-hydrazine hydrochloride. The title compound was synthesized according to the method described in U.S. Pat. No. 3,959,309 (1976). To a stirred solution of 2-bromo-5-fluoroaniline (1) (100 g, 0.526 mol; Aldrich) in diethyl ether (600 mL) was added drop-wise concentrated hydrochloric acid (44 mL). After stirring for another 20 min, the precipitate was filtered, washed thoroughly with diethyl ether, and dried in vacuo to obtain 94.7 g of 2-bromo-5-fluoroaniline hydrochloride as a white powder: HPLC rt=2.21 min; 1H NMR (DMSO-d6, 500 MHz) δ ppm 6.36 (1H, dt, J=8.5, 3 Hz, 5-CH), 76.67 (1H, dd, J=11, 3 Hz, 3-CH), 7.36 (1H, dd, J=9, 6 Hz, 6-CH).
To a mechanically stirred suspension of 2-bromo-5-fluoroaniline hydrochloride (94.7 g, 0.418 mol; obtained above) in a mixture of conc-HCl (250 mL) and water (250 mL) was added drop-wise a solution of sodium nitrite, NaNO2 (29 g, 0.420 mol) in water (60 mL) in a dry-ice/acetone bath, maintaining the bath temperature at −5-0° C. over a period of 10-15 min. Additional NaNO2 (1 g) was added and stirred at the same temperature for 5 min to obtain a clear solution. To this clear solution was added a solution of stannous chloride, SnCl2 (276.8 g, 1.46 mol) in conc-HCl (600 mL) drop-wise at 0-5° C. over a period of 30-40 min. A precipitate starts forming immediately after the addition. This thick mixture was stirred for another 1.5 h, and the precipitate was collected by suction-filtration, washed with a minimum amount of water ( ̃200 mL), then with diethyl ether, and dried in vacuo to obtain the title compound as an off-white powder: HPLC rt=0.73 min; LC/MS m/z 205 (M+H); 1H NMR (DMSO-d6, 500 MHz) δ ppm 6.78 (1H, dt, J=8.5, 3 Hz, 5-CH), 7.04 (1H, dd, J=11, 3 Hz, 3-CH), 7.60 (1H, dd, J=8.5, 6 Hz, 6-CH), 8.15 (1H, s, NH), 10.49 (3H, brs, NH3+); 13C NMR (DMSO-d6, 125.8 Hz) δ ppm 102.6 (d, J=28 Hz, 3-CH), 103.4 (d, J=2.6 Hz, 1-C), 109.0 (d, J=23 Hz, 5-CH), 133.8 (d, J=9.6 Hz, 6-CH), 144.0 (d, J=10.7 Hz, 2-C), 161.7 (d, J=243 Hz, 4-CF).
Intermediate 234




1-(2-Bromo-5-fluoro-phenyl)-1H-pyrazole. A mixture of 1-(2-bromo-5-fluoro-phenyl)-hydrazine hydrochloride (1.83 g, 7.58 mmol) and malonaldehyde bis(dimethylacetal) (1.27 g, 7.73 mmol) in EtOH (10 mL) was heated at reflux under nitrogen for 1 h. After cooling the mixture was diluted with diethyl ether (25 mL) and washed with water, and then with brine, dried (Na2SO4), filtered and concentrated to obtain the title compound as a clear oil: HPLC rt=2.31 min. LC/MS m/z 241/243 (M+H); 1H NMR (CDCl3, 500 MHz) δ ppm 6.47 (1H, m, 8-CH), 7.00-7.04 (1H, m, 5-CH), 7.31 (1H, dd, J=9, 3 Hz, 3-CH), 7.65 (1H, dd, J=8.8, 5.5 Hz, 6-CH), 7.75 (1H, d, J=1.5 Hz, 9-CH), 7.89 (1H, d, J=2.5 Hz, 7-CH); 13C NMR (CDCl3, 125.8 Hz) δ ppm 107.0 (8-CH), 112.2 (d, J=3.8 Hz, 1-C), 115.8, (d, J=25 Hz, 3-CH), 116.8 (d, J=22 Hz, 5-CH), 131.3 (7-CH), 134.9 (d, J=8.6 Hz, 6-CH), 140.9 (d, J=10 Hz, 2-C), 141.4 (9-CH), 161.9 (d, J=250 Hz, 4-CF); HRMS (ESI) calcd for C9H7BrN2 (M+H) 240.9777; found 240.9769 ( δ-3.1 ppm).
Intermediate 235




4-Fluoro-2-pyrazol-1-yl-benzonitrile. To a stirred suspension of CuCN (358 mg, 4.0 mmol, Aldrich) in DMF (2 mL, Sure Seal) was added a solution of 1-(2-Bromo-5-fluoro-phenyl)-1H-pyrazole (964 mg, 4.0 mmol) in DMF (2 mL) and the resulting clear solution was stirred in an oil bath heated at 110° C. under nitrogen for 1.5 h. After cooling, the mixture was concentrated in vacuo and the residue mixed with 14% aqueous NH4OH (10 mL) was stirred at room temperature for 0.5 h. To this was added EtOAc (15 mL) and the mixture stirred for additional 0.5 h. The EtOAc extract was washed with dil. NH4OH and then with brine, dried (Na2SO4), filtered, and concentrated. The residual oil was purified by column chromatography (SiO2, 0-10% EtOAc/CH2Cl2) to obtain 542 mg of the title compound as a white crystalline solid: TLC Rf 0.42 (CH2Cl2); HPLC rt=1.94 min. LC/MS m/z 188 (M+H); 1H NMR (CDCl3, 500 MHz) δ ppm 6.55 (1H, dd, J=2.6, 1.7 Hz, 9-CH), 7.10-7.13 (1H, m, 5-CH), 7.59 (1H, dd, J=9.5, 2.5 Hz, 3-CH), 7.76 (1H, dd, J=8.7, 5.5 Hz, 6-CH), 7.80 (1H, d, J=−1.2 Hz, 10-CH), 8.25 (1H, d, J=2.7 Hz, 8-CH); 13C NMR (CDCl3, 125.8 Hz) δ ppm 100.5 (d, J=3.8 Hz, 1-C), 109.1 (9-CH), 111.9 (d, J=27 Hz, 3-CH), 114.9 (d, J=23 Hz, 5-CH), 116.6 (7-CN), 129.5 (8-CH), 136.7 (d, J=10 Hz, 6-CH), 142.8 (10-CH), 144.2 (d, J=12 Hz, 2-C), 165.5 (d, J=258 Hz, 4-CF); HRMS (ESI) calcd for C10H7FN3 (M+H) 189.0576, found 189.0568 (δ-4.5 ppm); UV (MeOH) λ max 224 nm (ε2.54×104), 261 nm (ε1.20×104); Anal. calcd for C10H7FN3: C, 64.16; H, 3.24; N, 22.45; found C, 64.02; H, 3.16; N, 22.68.
Intermediate 236




4-Fluoro-2-pyrazol-1-yl-benzylamine hydrochloride. To a solution of 4-fluoro-2-pyrazol-1-yl-benzonitrile (500 mg, 2.67 mmol) in EtOH (10 mL) and EtOAc (10 mL) was added conc-HCl (0.25 mL, 3 mmol) and 10% Pd—C (100 mg). The mixture was hydrogenated at 1 atmosphere hydrogen for 20 h. To the mixture was added additional conc-HCl (0.15 mL) and 10% Pd—C (100 mg) and the mixture was further hydrogenated for additional 20 h. The mixture was filtered over Celite, washed with EtOH, and the combined filtrate and washings were concentrated in vacuo. The residual material was triturated with EtOH to obtain 356 mg of the title compound as the hydrochloride salt as an off-white crystalline solid: HPLC rt=1.01 min. LC/MS m/z 192 (M+H); 1H NMR (DMSO-d6, 500 MHz) δ ppm 3.99 (2H, s, 7-CH2), 6.63 (1H, t, J=2 Hz, 9-CH), 7.40 (1H, dt, J=8.5, 2.5 Hz, 5-CH), 7.54 (1H, dd, J=10, 2.5 Hz, 3-CH) 7.81 (1H, dd, J=8.5, 6 Hz, 6-CH), 7.86 (1H, d, J=1.5 Hz, 10-CH) 8.35 (1H, d, J=2 Hz, 8-CH), 8.54 (3H, br.s, NH3+); 13C NMR (DMSO-d6, 125.8 Hz) δ ppm 38.8 (7-CH2), 107.7 (9-CH2), 111.6 (d, J=25 Hz, 3-CH), 114.6 (d, J=21 Hz, 5-CH), 123.7 (d, J=3 Hz, 1-C), 131.7 (8-CH), 134.0 (d, J=9 Hz, 6-CH), 140.4 (d, J=11 Hz, 2-C), 141.5 (10-CH), 161.9 (d, J=247 Hz, 6-CF); HRMS (ESI) calcd for C10H11FN3 (M+H) 192.0937, found 192.0930 (δ-3.6 ppm).
Intermediate 237




1-(2-Bromo-5-fluorophenyl)-3,5-dimethyl-1H-1,2,4-triazole. A mixture of 1-(2-bromo-5-fluoro-phenyl)-hydrazine hydrochloride (24.15 g, 100 mmol) and diacetamide (10.1 g, 100 mmol; Aldrich) in anhydrous pyridine (100 mL) was stirred in an oil bath heated at 125-130° C. under nitrogen for 2 h. After cooling the mixture was concentrated in vacuo to dryness, and the residue diluted with EtOAc (100 mL) was washed with water (50 mL), and then with brine (30 mL), dried (Na2SO4), filtered and concentrated to obtain 24.2 g of the title compound as a brownish oil: HPLC rt=1.59 min. LC/MS m/z 270/272 (M+H); 1H NMR (CDCl3, 500 MHz) δ ppm 2.30 (3H, s, 9-Me), 2.39 (3H, s, 10-Me), 7.10-7.15 (2H, m, 3,5-CH), 7.67 (1H, dd, J=8.5, 5.5 Hz, 6-H); 13C NMR (CDCl3, 125.8 Hz) δ ppm 12.4 (9-CH3), 13.8 (10-Me), 116.5 (d, J=4 Hz, 1-C), 117.2, (d, J=24 Hz, 3-CH), 118.9 (d, J=22 Hz, 5-CH), 134.7 (d, J=8.5 Hz, 6-CH), 137.8 (d, J=10 Hz, 2-C), 153.9 (7-C), 160.9 (8-C), 161.8 (d, J=251 Hz, 4-CF); HRMS (ESI) calcd for C10H10BrFN3 (M+H) 270.0042, found 270.0048 (δ+2.2 ppm). This triazole was also prepared in 62% yield from 1-(2-bromo-5-fluoro-phenyl)-hydrazine hydrochloride and 2,4,6-trimethyl-s-triazine by refluxing them in EtOH.
Intermediate 238




2-(3,5-Dimethyl-1H-1,2,4-triazol-1-yl)-4-fluorobenzonitrile LC/MS m/z 217 (M+H). 1H NMR (CDCl3, 500 MHz) δ ppm 2.45 (3H, s, 11-Me), 2.49 (3H, s, 10-Me), 7.25 (1H, dd, J=8.5, 2.5 Hz, 3-CH), 7.31 (1H, dt, J=8.5, 2.5 Hz, 5-CH), 7.87 (1H, dd, J=8.7, 5.6 Hz, 6-CH). 13C NMR (CDCl3, 125.8 Hz) δ ppm 12.7 (10-Me), 13.8 (11-Me), 107.1 (d, J=3.8 Hz, 1-C), 114.8 (7-CN), 116.2, (d, J=25 Hz, 3-CH), 117.6 (d, J=22 Hz, 5-CH), 136.0 (d, J=10 Hz, 6-CH), 141.6 (d, J=10 Hz, 2-C), 153.9 (8-C), 161.0 (9-C), 165.0 (d, J=260 Hz, 4-CF). HRMS (ESI) calcd for C11H10FN4 (M+H) 217.0889, found 271.0879 (δ-4.8 ppm). Anal. calcd for C11H9FN4: C61.10, H4.19, N25.91; found C, 60.78; H, 3.93; N, 26.05.
Intermediate 239




(2-(3,5-Dimethyl-1H-1,2,4-triazol-1-yl)-4-fluorophenyl)methanamine dihydrochloride. LC/MS m/z 221 (M+H). 1H NMR (CD3OD, 500 MHz) δ ppm 2.67 (3H; s, 11-Me), 2.77 (3H, s, 10-Me), 4.16 (2H, s, 7-CH2), 7.61 (1H, dt, J=8.5, 2.5 Hz, 5-CH), 7.67 (1H, dd, J=8.5, 2.4 Hz, 3-CH), 7.93 (1H, dd, J=8.8, 5.7 Hz, 6-CH). 13C NMR (CD3OD, 125.8 MHz) δ ppm 10.5, 10.7 (10,11-Me), 38.2 (7-CH2), 115.1, (d, J=26 Hz, 3-CH), 119.3 (d, J=21 Hz, 5-CH), 126.9 (d, J=3.8 Hz, 1-C), 134.5 (d, J=9.6 Hz, 6-CH), 135.3 (d, J=110.6 Hz, 2-C), 154.1, 154.4 (8,9-C), 163.1 (d, J=251 Hz, 4-CF). HRMS (ESI) calcd for C11H14FN4 (M+H) 221.1202, found 221.1204 (δ-0.7 ppm); UV (MeOH) λ max 232 nm (8.74×103); Anal calcd for C11H13FN4.2.2HCl.0.6H2O: C42.47; H5.25; N18.01; C125.07; H2O3.48; found: 42.95, H4.80, N18.41, C124.56, H2O3.00 (KF).
Intermediate 240




1-(2-bromo-5-fluorophenyl)-1H-1,2,4-triazole. Prepared according to the procedure described in J. Org. Chem., 1956, 21, 1037. A stirred mixture of (2-bromo-5-fluoro-phenyl)-hydrazine hydrochloride (31.9 g, 0.132 mol) and s-triazine (10.7 g, 0.132 mol; Aldrich) in EtOH (250 mL) was heated at 90° C. for 3 h. After cooling, the insoluble material was filtered off and the filtrate was concentrated. The residue was partitioned between CH2Cl2 (200 mL) and water (200 mL), and the aqueous phase was extracted once more with CH2Cl2 and the combined extracts were dried (MgSO4), filtered and concentrated to a red semi-solid which was triturated with hexanes to give 19.6 g of the title compound as an orange solid: HPLC rt=1.68 min. LC/MS m/z 242 (M+H); 1H NMR (CDCl3, 500 MHz) δ ppm 7.02-7.15 (1H, m, 5-CH), 7.29 (1H, dd, J=8.4, 2.9 Hz, 3-CH), 7.68 (1H, dd, J=8.8, 5.5 Hz, 6-CH), 8.10 (1H, s, 8-CH), 8.54 (1H, s, 7-CH); 13C NMR (CDCl3, 125.8 Hz) δ ppm 112.54, 112.57 (d, J=1.5 Hz, 1-C), 115.68, 115.88 (d, J=25 Hz, 3-CH), 118.07, 118.25 (d, J=22 Hz, 5-CH), 135.11, 135.18 (d, J=8.6 Hz, 6-CH), 137.43, 137.52 (d, J=11 Hz, 2-C), 144.4 (7-CH), 152.6 (8-CH), 160.95, 162.94 (d, J=251 Hz, 4-CF); HRMS (ESI) calcd for C8H6BrN3 (M+H) 241.9727; found 241.9733 (δ 1.6 ppm); UV (MeOH) λmax 275 nm (ε1.45×103); Anal. calcd for C8H5BrFN3: C, 39.69; H, 2.08; N, 17.36; found C, 39.63; H, 1.83; N, 17.22.
Intermediate 241




(2-(2-Bromo-5-fluorophenyl)-2H-1,2,4-triazol-3-yl)methanol. A suspension of 1-(2-bromo-5-fluorophenyl)-1H-1,2,4-triazole (3.63 g, 15 mmol) in 37% formaline (15 mL) in a sealed vessel was stirred in an oil bath heated at 150° C. for 9.5 h. After cooling, the mixture was extracted with CH2Cl2 (15 mL×2) and the combined extracts were washed with brine, dried (Na2SO4), filtered and concentrated. The residual crude oil was purified by column chromatography (SiO2, 0-7% MeOH/CH2C12) followed by trituration with Et2O to obtain 430 mg of the title compound as white crystalline solid: TLC Rf 0.5 (10% MeOH/CH2Cl2); HPLC rt=1.31 min; LC/MS m/z 272/274 (M+H); 1H NMR (CDCl3, 500 MHz) δ ppm 3.67 (1H, br.s, OH), 4.65 (2H, s, 9-OCH2), 7.13-7.22 (1H, m, 5-CH), 7.29 (1H, dd, J=8, 3 Hz, 3-CH), 7.71 (1H, dd, J=9.0, 5.3 Hz, 6-CH), 8.03 (1H, s, 8-CH); 13C NMR (CDCl3, 125.8 Hz) δ ppm 55.3 (9-OCH2), 116.2 (d, J=3.9 Hz, 1-C), 117.4 (d, J=25 Hz, 3-CH), 119.4 (d, J=22 Hz, 5-CH), 134.7 (d, J=8.3 Hz, 6-CH), 137.0 (d, J=10 Hz, 2-C), 151.3 (8-CH), 156.3 (7-C), 161.8 (d, J=252 Hz, 4-CF); HRMS (ESI) calcd for C9H8BrN3O (M+H) 271.9835; found 271.9828 (δ-2.5 ppm).
Intermediate 242




4-Fluoro-2-(5-hydroxymethyl-[1,2, 4]triazol-1-yl)-benzonitrile. This compound was prepared from (2-(2-bromo-5-fluorophenyl)-2H-1,2,4-triazol-3-yl)methanol and CuCN in DMF at 125-130° C. for 5 h by the method used for the preparation of 4-fluoro-2-pyrazol-1-yl-benzonitrile. Yield: 48% (beige solid) HPLC rt=1.11 min. LC/MS m/z 219 (M+H). 1H NMR (CDCl3, 500 MHz) δ ppm 2.39 (1H, br.s, OH), 4.79 (2H, s, 9-OCH2), 7.32-7.40 (1H, m, 5-CH), 7.50 (1H, d, J=8.2, 2.1 Hz, 3-CH), 7.87 (1H, dd, J=8.5, 5.5 Hz, 6-CH), 8.14 (1H, s, 9-CH). HRMS (ESI) calcd for C10H8FN4O (M+H) 219.0682; found 219.0689 (δ+3.1 ppm).
Intermediate 243




[2-(2-Aminomethyl-5-fluoro-phenyl)-2H-[1, 2, 4]triazol-3-yl]-methanol hydrochloride. This compound was prepared from 4-fluoro-2-(5-hydroxymethyl-[1,2,4]triazol-1-yl)-benzonitrile by the hydrogenation method used for the preparation of 4-fluoro-2-pyrazol-1-yl-benzylamine hydrochloride. Yield: 100% (grayish green powder). HPLC rt=0.33 min. LC/MS m/z 223 (M+H). 1H NMR (DMSO-d6, 500 MHz) δ ppm 3.76 (2H, q, J=5.8 Hz, 7-NCH2), 4.55 (2H, s, 9-OCH2), 7.54-7.60 (1H, m, 5-CH), 7.62 (1H, dd, J=9, 2.6 Hz, 3-CH), 7.89 (1H, dd, J=8.7, 6.0 Hz, 6-CH), 8.26 (1H, s, 8-CH), 8.58 (3H, br.s, NH3). 13C NMR (DMSO-d6, 125.8 Hz) δ ppm 37.1 (7-NCH2), 54.2 (10-OCH2), 114.6 (d, J=24 Hz, 3-CH), 117.0 (d, J=21 Hz, 5-CH), 127.1 (d, J=3.8 Hz, 1-C), 132.2 (d, J=8.6 Hz, 6-CH), 136.9 (d, J=11 Hz, 2-C), 151.0 (9-CH), 156.3 (8-C), 161.3 (d, J=247 Hz, 4-CF). HRMS (ESI) calcd for C10H12FN4O (M+H) 223.0995; found 223.0988 (δ-3.2 ppm).
Example 1




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. A mixture of intermediate 7, ethyl 3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.036 g, 0.15 mmol) and 4-fluorobenzylamine (0.11 g, 0.87 mmol) in anhydrous ethyl alcohol (5 ml) and N,N-dimethylformamide (2 ml) was heated under reflux for 18 h. The solvent was then evaporated in vacuo and the residue was partitioned between ethyl acetate and 0.1 N hydrochloric acid. The organic phase was washed with water, brine and dried over anhydrous sodium sulfate. Evaporation of the solvent and recrystallization of the resulting solid from ethanol gave 0.023 g (47% yield) of the title amide as white crystals; mp 211° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ ppm: 4.06 (4H, m, 2×CH2), 4.59 (2H, d, J=7.6 Hz, NCH2), 4.61 (2H, s, OCH2), 7.09 (2H, m, aromatics), 7.33 (2H, m, aromatics), 7.84 (1H, broad t, NH), 12.06 (1H, s, OH). MS (ESI+) m/z 320 [M+H+].
Example 2




Pyrimido[2,1-c[]1,4]oxazine-2-carboxamide, N-[(4-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. Reaction of intermediate 7, ethyl 3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.100 g, 0.42 mmol) with 4-fluoro-3-methylbenzylamine (0.23 g, 1.66 mmol) as described in the preparation of example 1 gave 0.101 g (73% yield) of the title amide as white crystals; mp 206-208° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ ppm: 2.30 (3H, s, CH3), 4.06 (4H, m, 2×CH2), 4.55 (2H, d, J=6.1 Hz, NCH2), 4.60 (2H, s, OCH2), 7.01 (1H, m, aromatic), 7.14 (2H, m, aromatics), 7.81 (1H, broad t, NH), 12.09 (1H, s, OH). MS (ESI+) m/z 334 [M+H+].
Example 3




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(methylamino)carbonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from intermediate 150, N-(4-fluoro-2-(methylcarbamoyl)benzyl)-3-(benzyloxy)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (DMSO-d6) δ ppm: 2.79 (3H, d, J=4.5 Hz, NCH3), 3.83 (2H, m, CH2), 4.02 (2H, m, CH2), 4.54 (2H, d, J=6.7 Hz, NCH2), 4.58 (2H, s, OCH2), 7.31 (2H, m, aromatics), 7.38 (1H, m, aromatic), 8.54 (1H, broad q, NH), 9.21 (1H, broad t, NH), 12.24 (1H, s, OH). HRMS (ESI+) calculated for C17H18FN4O5 [M+H+]: 377.1261; found: 377.1249.
Example 4




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. Reaction of intermediate 15, ethyl 3-hydroxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate (0.050 g, 0.20 mmol) with 4-fluorobenzylamine (0.11 g, 0.87 mmol) as described in the preparation of example 1 gave 0.056 g (84% yield) of the title amide as white crystals; mp 165-167° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ ppm: 1.62 (3H, d, J=7.0 Hz, CH3), 3.90 (2H, m, CH2), 4.15-4.32 (2H, m, CH2), 4.61 (3H, m, NCH2 and OCH), 7.08 (2H, m, aromatics), 7.34 (2H, m, aromatics), 7.82 (1H, broad t, NH), 12.06 (1H, s, OH). Anal. Calcd for C16H16FN3O4: C, 57.65; H, 4.83; N, 12.60. Found: C, 57.44; H, 4.69; N, 12.37.
Example 5




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. Reaction of intermediate 15, ethyl 3-hydroxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.090 g, 0.35 mmol) with 4-fluoro-3-methylbenzylamine (0.180 g, 1.3 mmol) as described in the preparation of example 1 gave 0.068 g (55% yield) of the title amide as white crystals; mp 134° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ ppm: 1.62 (3H, d, J=6.6 Hz, CH3), 2.30 (3H, s, CH3), 3.90 (2H, m, CH2), 4.13-4.32 (2H, m, CH2), 4.49-4.64 (3H, m, NCH2 and OCH), 7.01 (1H, m, aromatic), 7.16 (2H, m, aromatics), 7.79 (1H, broad t, NH), 12.09 (1H, s, OH). Anal. Calcd for C17H18FN3O4: C, 58.78; H, 5.22; N, 12.09. Found: C, 58.57; H, 5.55; N, 11.90.
Example 6




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,4-dichlorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. Reaction of intermediate 15, ethyl 3-hydroxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate (0.075 g, 0.29 mmol) with 3,4-dichlorobenzylamine (0.140 g, 0.8 mmol) as described in the preparation of example 1 gave 0.085 g (75% yield) of the title amide as white crystals; mp 192° C. (ethyl acetate-hexane). 1HNMR 400 MHz (CDCl3) δ ppm: 1.64 (3H, d, J=6.6 Hz, CH3), 3.91 (2H, m, CH2), 4.17-4.32 (2H, m, CH2), 4.50-4.68 (3H, m, NCH2 and OCH), 7.20 (1H, dd, J=2.0 Hz and J=8.0 Hz, aromatic), 7.44 (1H, d, J=2.0 Hz, aromatic), 7.46 (1H, d, J=8.0 Hz, aromatic), 7.86 (1H, broad t, NH), 11.92 (1H, s, OH). MS (ESI+) m/z 384 [M+H+]. Anal. Calcd for C16H15Cl2N3O4: C, 50.02; H, 3.94; N, 10.94. Found: C, 49.40; H, 4.06; N, 10.41.
Example 7




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3-chloro-4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. Reaction of intermediate 15, ethyl 3-hydroxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.075 g, 0.30 mmol) with 3-chloro-4-fluorobenzylamine (0.14 g, 0.88 mmol) as described in the preparation of example 1 gave 0.050 g (46% yield) of the title amide as white crystals; mp 172° C. (ethyl acetate-ether). 1HNMR 400 MHz (CDCl3) δ ppm: 1.63 (3H, d, J=6.6 Hz, CH3), 3.91 (2H, m, CH2), 4.17-4.32 (2H, m, CH2), 4.50-4.67 (3H, m, NCH2 and OCH), 7.16 (1H, m, aromatic), 7.24 (1H, m, aromatic), 7.40 (1H, m, aromatic), 7.85 (1H, broad t, NH), 11.95 (1H, s, OH). Anal. Calcd for C16H15ClFN3O4: C, 52.25; H, 4.11; N, 11.42. Found: C, 51.99; H, 4.01; N, 11.09.
Example 8




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,4-dimethylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. Reaction of intermediate 15, ethyl 3-hydroxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.075 g, 0.30 mmol) with 3,4-dimethylbenzylamine (0.15 g, 1.1 mmol) as described in the preparation of example 1 gave 0.033 g (33% yield) of the title amide as a white solid. 1HNMR 400 MHz (CDCl3) δ ppm: 1.61 (3H, d, J=6.6 Hz, CH3), 2.28 (3H, s, CH3), 2.29 (3H, s, CH3), 3.90 (2H, m, CH2), 4.16-4.30 (2H, m, CH2), 4.50-4.65 (3H, m, NCH2 and OCH), 7.08-7.17 (3H, m, aromatics), 7.78 (1H, broad t, NH), 12.17 (1H, s, OH). MS (ESI+) m/z 344 [M+H+].
Example 9




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(methylamino)carbonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. A solution of intermediate 140, N-(4-fluoro-2-(methylcarbamoyl)benzyl)-3-benzyloxy-9-methyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, (0.268 g, 0.56 mmol) in a mixture of ethyl acetate (25 ml) and ethanol (25 ml) was hydrogenated under 1 atm of hydrogen at 25° C. over 10% palladium on activated carbon (0.09 g) for 2.5 h to give 0.121 g (56% yield) of the title ester as a white solid. 1HNMR 400 MHz (DMSO-d6) δ ppm: 1.57 (3H, d, J=6.7 Hz, CH3), 2.79 (3H, d, J=4.6 Hz, NCH3), 3.70 (1H, m, CH), 3.87 (1H, m, CH), 3.98 (1H, m, CH), 4.15 (1H, m, CH), 4.55 (2H, m, NCH2), 4.62 (1H, q, J=6.6 Hz, OCH), 7.25-7.44 (3H, m, aromatics), 8.59 (1H, broad q, NH), 9.39 (1H, broad, NH), 12.18 (1H, s, OH). HRMS (ESI+) calculated for C18H20FN4O5 [M+H+]: 391.1418; found: 391.1431.
Examples 10-14
Examples 10-14 can be prepared from ethyl 9-ethyl-3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and the indicated amines according to the method described for the synthesis of example 1. Ethyl 9-ethyl-3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate was prepared according to the method used to prepare intermediate 15.
Example 10




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9-ethyl-N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from 4-fluorobenzylamine. 1HNMR 400 MHz (CDCl3) δ ppm: 1.02 (3H, t, J=7.3 Hz, CH3), 1.93 (1H, m, CH), 2.15 (1H, m, CH), 3.88 (2H, m, CH2), 4.2-4.29 (2H, m, CH2), 4.46 (1H, m, CH), 4.53-4.69 (2H, m, CH2), 7.06 (2H, m, aromatics), 7.34 (2H, m, aromatics), 7.82 (1H, broad t, NH), 12.05 (1H, s, OH). HRMS (ESI+) calculated for C17H19FN3O4 [M+H+]: 348.1360; found: 348.1355.
Example 11




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9-ethyl-N-[(4-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from (4-fluoro-3-methylphenyl)methanamine. 1HNMR 400 MHz (CDCl3) δ ppm: 1.01 (3H, t, J=7.4 Hz, CH3), 1.91 (1H, m, CH), 2.16 (1H, m, CH), 2.30 (3H, s, CH3), 3.87 (2H, m, CH2), 4.2-4.30 (2H, m, CH2), 4.46 (1H, m, CH), 4.48-4.65 (2H, m, CH2), 7.01 (1H, m, aromatic), 7.14 (2H, m, aromatics), 7.81 (1H, broad t, NH), 12.07 (1H, s, OH). Anal. Calcd for C18H20FN3O4: C, 59.82; H, 5.57; N, 11.62; Found: C, 59.53; H, 5.86; N, 11.42.
Example 12




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,4-dichlorophenyl)methyl]-9-ethyl-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from (3,4-dichlorophenyl)methanamine. 1HNMR 400 MHz (CDCl3) δ ppm: 1.00 (3H, t, J=7.3 Hz, CH3), 1.91 (1H, m, CH), 2.15 (1H, m, CH), 3.85 (2H, m, CH2), 4.19-4.28 (2H, m, CH2), 4.45 (1H, m, CH), 4.48-4.66 (2H, m, CH2), 7.19 (1H, m, aromatic), 7.43 (2H, m, aromatics), 7.84 (1H, broad t, NH), 11.88 (1H, s, OH). Anal. Calcd for C17H17Cl2N3O4: C, 51.27; H, 4.30; N, 10.55; Found: C, 51.16; H, 4.21; N, 10.34.
Example 13




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,4-dimethylphenyl)methyl]-9-ethyl-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from (3,4-dimethylphenyl)methanamine. 1HNMR 400 MHz (CDCl3) δ ppm: 0.98 (3H, t, J=7.3 Hz, CH3), 1.87 (1H, m, CH), 2.12 (1H, m, CH), 2.26 (3H, s, CH3), 2.27 (3H, s, CH3), 3.83 (2H, m, CH2), 4.17-4.26 (2H, m, CH2), 4.41 (1H, m, CH), 4.45-4.64 (2H, m, CH2), 7.05-7.24 (3H, m, aromatics), 7.75 (1H, broad t, NH), 12.13 (1H, s, OH). Anal. Calcd for C19H23N3O4: C, 63.85; H, 6.49; N, 11.76; Found: C, 63.55; H, 6.48; N, 11.74.
Example 14




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3-chloro-4-fluorophenyl)methyl]-9-ethyl-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from (3-chloro-4-fluorophenyl)methanamine. 1HNMR 400 MHz (CDCl3) δ ppm: 1.03 (3H, t, J=7.3 Hz, CH3), 1.92 (1H, m, CH), 2.17 (1H, m, CH), 3.88 (2H, m, CH2), 4.18-4.32 (2H, m, CH2), 4.46 (1H, m, CH), 4.5-4.68 (2H, m, CH2), 7.16 (1H, m, aromatic), 7.24 (1H, m, aromatic), 7.40 (1H, m, aromatic), 7.84 (1H, broad t, NH), 11.93 (1H, s, OH). Anal. Calcd for C17H17ClFN3O4: C, 53.48; H, 4.48; N, 11.00; Found: C, 53.25; H, 4.49; N, 10.79.
Examples 15-16
Examples 15-16 can be prepared from 3-(benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid and the indicated amines according to the methods described for the synthesis of intermediate 140 and example 9. 3-(Benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylic acid was prepared according to the method used to prepare intermediate 16.
Example 15




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9-ethyl-N-[[4-fluoro-2-[(methylamino)carbonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from intermediate 39.1HNMR 400 MHz (CDCl3) δ ppm: 1.03 (3H, t, J=7.3 Hz, CH3), 1.98 (1H, m, CH), 2.28 (1H, m, CH), 3.06 (3H, d, J=4.5 Hz, NCH3), 3.86 (2H, m, CH2), 4.14-4.29 (2H, m, CH2), 4.48 (1H, m, CH), 4.60 (2H, m, CH2), 6.2 (1H, broad, NH), 7.14-7.21 (2H, m, aromatics), 7.54 (1H, m, aromatic), 8.85 (1H, broad t, NH), 12.1 (1H, s, OH). HRMS (ESI+) calculated for C19H22FN4O5 [M+H+]: 405.1574; found: 405.1579.
Example 16




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9-ethyl-N-[[4-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from intermediate 151, N-(4-fluoro-2-(1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9-ethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, according to a procedure similar to that described in example 9. 1HNMR 400 MHz (DMSO-d6) δ ppm: 0.94 (3H, t, J=7.5 Hz, CH3), 1.88 (1H, m, CH), 2.27 (1H, m, CH), 3.68 (1H, m, CH), 3.87 (1H, m, CH), 4.0 (1H, m, CH), 4.19 (1H, m, CH), 4.36-4.49 (3H, m, CH2 and CH), 7.43 (1H, m, aromatic), 7.56 (2H, m, aromatics), 8.32 (1H, m, CH), 9.05 (1H, m, CH), 9.3 (1H, broad t, NH), 12.04 (1H, s, OH). HRMS (ESI+) calculated for C19H20FN6O4 [M+H+]: 415.1530; found: 415.1515.
Examples 17-18
Examples 17-18 can be prepared from ethyl 3-hydroxy-9-isopropyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and the indicated amines according to the method described for the synthesis of example 1. Ethyl 3-hydroxy-9-isopropyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate can be prepared according to the method used to prepare intermediate 15.
Example 17




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-(1-methylethyl)-4-oxo-. The title compound can be prepared from 4-fluorobenzylamine. 1HNMR 400 MHz (CDCl3) δ ppm: 0.80 (3H, d, J=6.7 Hz, CH3), 1.13 (3H, d, J=7.1 Hz, CH3), 2.54 (1H, m, CH), 3.77 (2H, m, CH2), 4.3 (2H, m, CH2), 4.40 (1H, d, J=2.5 Hz, CH), 4.50-4.72 (2H, m, CH2), 7.09 (2H, m, aromatics), 7.34 (2H, m, aromatics), 7.82 (1H, broad t, NH), 12.04 (1H, s, OH). Anal. Calcd for C18H20FN3O4: C, 59.82; H, 5.57; N, 11.62; Found: C, 59.22; H, 5.81; N, 11.50.
Example 18




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-(1-methylethyl)-4-oxo-. The title compound can be prepared from (4-fluoro-3-methylphenyl)methanamine. 1HNMR 400 MHz (CDCl3) δ ppm: 0.80 (3H, d, J=6.6 Hz, CH3), 1.13 (3H, d, J=7.1 Hz, CH3), 2.30 (3H, s, CH3), 2.54 (1H, m, CH), 3.77 (2H, m, CH2), 4.3 (2H, m, CH2), 4.40 (1H, broad s, CH), 4.46-4.68 (2H, m, CH2), 7.02 (1H, m, aromatic), 7.17 (2H, m, aromatics), 7.80 (1H, broad t, NH), 12.07 (1H, s, OH). HRMS (ESI+) calculated for C19H23FN3O4 [M+H+]: 376.1673; found: 376.1671.
Example 19




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. To a solution of intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (3.0 g, 11.19 mmol) in DMF (20 mL) and ethanol (10 mL) was added triethylamine (1.55 mL) followed by 4-fluorobenzylamine (3.82 mL, 33.57 mmol). The mixture was stirred at 90° C. for 2 h and then concentrated. The resultant oil was partitioned between ethyl acetate (50 mL) and 1N aqueous HCl (35 mL). The aqueous layer was back-extracted with ethyl acetate (20 mL) and the organic layers were combined and washed with H2O (4×20 mL) and brine then dried (Na2SO4) and concentrated. The brown residue was triturated with ether and the solids filtered and washed with ether. The pale brown solids were recrystallized from 95:5 MeOH/H2O to give the title compound as colorless needles (3.18 g, 82% yield). 1H NMR (500 MHz, CDCl3) δ ppm: 11.96 (1H, s), 7.77 (1H, brs), 7.30 (2H, dd, J=8.4, 5.3 Hz), 7.04 (2H, t, J=8.7 Hz), 4.57 (2H, d, J=6.1 Hz), 4.01 (4H, s), 1.56 (6H, s). HRMS (M+H) calcd for C17H19FN3O4: 348.13597; found: 348.1365. Anal calcd for C17H18FN3O4: C, 58.78; H, 5.22; N, 12.09. Found: C, 58.38; H, 5.23; N, 11.80.
Additional procedure. Ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate (50 g, 0.186 mol) and ethanol SDA3A (500 mL) was added to a 1 L round bottom flask equipped with a mechanical stirrer, nitrogen inlet-outlet, temperature probe, and condenser. Triethylamine (18.86 g, 0.186 mol) was added to the slurry. The resulting solution was heated to 35° C. followed by slow addition of 4-fluorobenzylamine (35.0 g, 0.279 mol). The thick slurry was heated at 78° C. with stirring for 6 h. The reaction mixture was polish filtered and the filtrate reheated to 70° C. The pH was adjusted to about 1.0 by slow addition of 1N HCl (405 mL). The thick slurry was cooled to 20-25° C., the solids filtered off, washed twice with ethanol SDA 3A (each 250 mL) and twice with water (each 500 mL). The white solid was dried in vacuo at 50-55° C. to afford N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide (58.9 g).
Additional procedure. To a stirring mixture of ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate (5.18 g, 19.31 mmol) and EtOH SDA3A (52 mL) was added triethylamine (1.95 g, 19.30 mmoles) under inert atmosphere. The resulting solution was heated to 40° C. and 4-fluorobenzylamine (3.58 g, 28.61 mmol) was added slowly over about 3 min. The resulting white slurry was heated to 78-79° C. giving a solution which was held with stirring for 6 h. The hot solution was polish filtered. The pH of the filtrate was adjusted to about 2.5 by slow addition of 1M H2SO4 (18.1 mL) at 65-70° C. The resulting thick slurry was allowed to cool slowly to 20-25° C. The solid was collected by filtration, washed with EtOH SDA 3A (2×25 mL) and water (2×25 mL). The solid was dried in vacuo at 50° C. to afford N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide as a white crystalline solid (6.02 g).
Example 20




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. A DMF solution of intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (4 mL, 1 mmol), Et3N (0.14 mL, 1 mmol) and 3-methyl-4-fluorobenzylamine (0.418 g, 3 mmol) was heated at 90° C. for 5 h. The reaction mixture was cooled and the product isolated by reverse phase preparative HPLC using MeOH/H2O-0.1% CF3CO2H as eluent. The fractions containing the desired material were combined and concentrated to afford the title compound as a yellow powder (0.19 g, 52% yield). 1H NMR (500 MHz, CDCl3) δ ppm: 11.99 (1H, s), 7.73 (1H, s), 7.14 (1H, d, J=7.3 Hz), 7.12-7.09 (1H, m), 6.98 (1H, t, J=9.0 Hz), 4.54 (2H, d, J=6.4 Hz), 4.01 (4H, s), 2.27 (3H, s), 1.56 (6H, s). HRMS (M+H) calcd for C18H21FN3O4: 362.1516; found: 362.1509. Anal calcd for C18H20FN3O4+0.07H2O: C, 59.26; H, 5.55; N, 11.48. Found: C, 58.88; H, 5.36; N, 11.34.
Examples 21-41
Examples 21-41 can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, and the indicated amines according to the method described for the synthesis of examples 1, 19 and 20.
Example 21




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 69. Solid, 1H NMR (500 MHz, DMSO-d6) δ ppm: 11.93 (1H, s), 9.29 (1H, t, J=6.2 Hz), 9.05 (1H, s), 8.32 (1H, s), 7.57-7.53 (2H, m), 7.43 (1H, td, J=7.9, 1.7 Hz), 4.44 (2H, d, J=6.1 Hz), 3.98 (2H, t, J=4.9 Hz), 3.83 (2H, t, J=4.9 Hz), 1.56 (6H, s). HRMS (M+H) calcd for C19H20N6O4F: 415.15302; found: 415.1520. Anal Calcd for C19H19N6O4F: C, 55.07; H, 4.62; N, 20.28; F, 4.58; found: C, 54.95; H, 4.67; N, 20.27; F, 4.56.
Example 22




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-fluoro-4-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 70. Solid, 1H NMR (500 MHz, DMSO-d6) δ ppm: 12.07 (1H, s), 9.46 (1H, bs), 9.33 (1H, s), 8.26 (1H, s), 7.81 (1H, dd, J=11.1, 2.0 Hz), 7.73 (1H, dd, J=8.2, 1.8 Hz), 7.52 (1H, t, J=8.2 Hz), 4.59 (2H, d, J=6.1 Hz), 3.98 (2H, t, J=5.0 Hz), 3.84 (2H, t, J=5.0 Hz), 1.58 (6H, s). HRMS (M+H) calcd for C19H20N6O4F: 415.15302; found: 415.1520. Anal calcd for C19H19N6O4F+1.25H2O: C, 52.23; H, 4.96; N, 19.23; F, 4.35; found: C, 52.29; H, 4.66; N, 19.23; F, 4.35.
Example 23




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(4-morpholinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 73. Solid, 1H NMR (500 MHz, CDCl3) δ ppm: 12.09 (1H, s), 7.91 (1H, brs), 7.31-7.28 (1H, m), 6.90 (1H, dd, J=10.4, 2.4 Hz), 6.84 (1H, td, J=8.1, 2.4 Hz), 4.66 (2H, d, J=6.4 Hz), 4.02 (4H, s), 3.89-3.87 (4H, m), 2.94-2.92 (4H, m), 1.59 (6H, s). HRMS (M−H) calcd for C21H24N4O5F: 431.17307; found: 431.1719. Anal calcd for C21H25N4O5F: C, 58.32; H, 5.82; N, 12.95; F, 4.39; found: C, 58.13; H, 5.81; N, 12.79; F, 4.33.
Example 24




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-fluoro-4-(4-morpholinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 74. 1H NMR (500 MHz, CDCl3) δ ppm: 12.01 (1H, s), 7.80 (1H, brs), 7.28-7.24 (1H, m), 6.66 (1H, d, J=8.5 Hz), 6.61 (1H, dd, J=13.4, 1.8 Hz), 4.56 (2H, d, J=6.1 Hz), 4.01 (4H, s), 3.85-3.84 (4H, m), 3.17-3.15 (4H, m), 1.60 (6H, s). HRMS (M−H) calcd for C21H24N4O5F: 431.17307; found: 431.1729. Anal calcd for C21H25N4O5F: C, 58.32; H, 5.82; N, 12.95; F, 4.39; found: C, 58.23; H, 5.73; N, 12.82; F, 4.21.
Example 25




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[3-(3,4-dichlorophenyl)propyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from 3-(3,4-dichlorophenyl)propan-1-amine. 1H NMR (500 MHz, CDCl3) δ ppm: 12.05 (1H, s), 7.46 (1H, brs), 7.35 (1H, d, J=8.2 Hz), 7.28 (1H, d, J=2.1 Hz), 7.04 (1H, dd, J=−8.2, 2.1 Hz), 4.02 (4H, s), 3.46 (2H, q, J=6.9 Hz), 2.67 (2H, t, J=7.6 Hz), 1.95 (2H, m), 1.59 (6H, s). HRMS (M+H) calcd for C19H22N3O4Cl2: 426.09875; found: 426.0996. Anal calcd for C19H21N3O4Cl2: C, 53.53; H, 4.96; N, 9.85; Cl, 16.63; found: C, 53.57; H, 4.96; N, 9.76; Cl, 16.63.
Example 26




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[3-(4-fluorophenyl)propyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from 3-(4-fluorophenyl)propan-1-amine. 1H NMR (500 MHz, CDCl3) δ ppm: 12.09 (1H, s), 7.15 (2H, dd, J=8.2, 5.5 Hz), 6.97 (2H, t, J=8.5 Hz), 4.02 (4H, s), 3.44 (2H, q, J=13.9, 6.9 Hz), 2.68 (2H, t, J=7.6 Hz), 1.98-1.92 (2H, m), 1.62 (6H, s). HRMS (M+H) calcd for C19H23N3O4F: 376.16727; found: 376.1687. Anal calcd for C19H22N3O4F: C, 60.79; H, 5.90; N, 11.19; F, 5.06; found: C, 60.70; H, 5.87; N, 11.14; F, 4.92.
Example 27




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(tetrahydro-1,1-dioxido-2H-1,2-thiazin-2-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 76. 1H NMR (500 MHz, CDCl3) δ ppm: 12.07 (1H, s), 8.31 (1H, d, J=6.44 Hz), 7.46 (1H, dd, J=8.5, 6.4 Hz), 7.19 (1H, dd, J=9.0, 2.6 Hz), 7.08 (1H, td, J=8.2, 2.7 Hz), 4.92 (1H, dd, J=14.0, 8.8 Hz), 4.37 (1H, dd, J=14.0, 3.4 Hz), 4.02-3.97 (2H, m), 3.99 (2H, s), 3.87-3.82 (1H, m), 3.45-3.41 (1H, m), 3.30-3.20 (2H, m), 2.46-2.32 (2H, m), 2.00-1.88 (2H, m), 1.57 (3H, s), 1.53 (3H, s). HRMS (M−H) calcd for C21H26N4O6FS: 481.15572; found: 481.1570. Anal calcd for C21H25N4O6FS: C, 52.49; H, 5.24; N, 11.66; F, 3.95; S, 6.67; found: C, 52.29; H, 5.37; N, 11.40; F, 3.91; S, 6.70.
Example 28




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,5-difluoro-2-pyridinyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 102. 1H NMR (500 MHz, CDCl3) δ ppm: 11.86 (1H, s), 8.58 (1H, brs), 8.33 (1H, d, J=2.4 Hz), 7.27-7.24 (1H, m), 4.76 (2H, d, J=5.2 Hz), 4.03 (4H, s), 1.63 (6H, s). HRMS (M+H) calcd for C16H17F2N4O4: 367.1218; found: 367.1230.
Example 29




N-((5-Chloropyridin-2-yl)methyl)-3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 103. 1H NMR (500 MHz, CDCl3) δ ppm: 11.80 (1H, brs), 8.64 (1H, brs), 8.57 (1H, d, J=2.4 Hz), 7.78 (1H, dd, J=8.6, 2.4 Hz), 7.44 (1H, d, J=8.6 Hz), 4.72 (2H, d, J=6.1 Hz), 4.02 (4H, s), 1.61 (6H, s). HRMS (M+H) calcd for C17H18ClN4O4: 365.1017; found: 365.1028. Anal calcd for C16H17ClN4O4.0.25H2O.0.5 CF3CO2H: C, 47.90; H, 4.26; N, 13.14, Cl, 8.32; found: C, 47.88; H, 3.98; N, 12.94, Cl, 8.57.
Example 30




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3-bromo-4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (3-bromo-4-fluorophenyl)methanamine Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 11.88 (1H, s), 7.76-7.84 (1H, br), 7.53 (1H, dd, J=6.3, 2.0 Hz), 7.26-7.29 (H, m), 7.07-7.14 (1H, m), 4.57 (2H, d, J=6.4 Hz), 4.02 (4H, s), 1.58 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.41, 157.79, 151.99, 146.52, 134.91, 132.89, 128.44, 128.39, 125.30, 117.04, 116.86, 77.69, 75.84, 58.21, 43.22, 41.98, 28.11. HRMS [M+H]+ calcd for C17H18N3O4FBr: 426.04648; found: 426.0468.
Example 31




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,4-dimethylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (3,4-dimethylphenyl)methanamine Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 7.74 (1H), 7.04-7.15 (3H, m), 4.56 (2H, d, J=5.8 Hz), 4.00-4.07 (4H, m), 2.27 (3H, s), 2.26 (3H, s), 1.57 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 167.92, 158.58, 151.61, 146.22, 137.37, 136.47, 134.53, 130.23, 129.11, 126.12, 125.12, 75.93, 58.10, 43.42, 43.00, 28.04, 19.85, 19.53. HRMS [M+H]+ calcd for C19H24N3O4: 358.17669; found: 358.1783.
Example 32




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3-chloro-4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (3-chloro-4-fluorophenyl)methanamine. Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 11.88 (1H, br s), 7.80 (1H, t, J=5.5 Hz), 7.38 (1H, dd, J=6.7, 2.1 Hz), 7.19-7.23 (1H, m), 7.13 (1H, t, J=8.5 Hz), 4.57 (2H, d, J=6.4 Hz), 4.02 (4H, s), 1.58 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.41, 157.82, 151.99, 146.52, 134.54, 130.02, 127.58, 127.52, 125.32, 121.61, 117.18, 117.01, 75.84, 58.21, 43.23, 42.08, 28.11. HRMS [M+H]+ calcd for C17H18N3O4FCl: 382.09644; found: 382.0980.
Example 33




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,4-difluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (3,4-difluorophenyl)methanamine. Light brown solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 7.77 (1H), 7.30-7.36 (2H, m), 7.27 (1H, s), 4.59 (2H, d, J=6.4 Hz), 4.01-4.06 (4H, m), 1.58 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.22, 158.36, 151.98, 151.17, 146.35, 134.33, 125.74, 123.70, 123.67, 117.87, 117.73, 116.88, 116.73, 75.90, 58.13, 43.42, 42.28, 28.06. HRMS [M+H]+ calcd for C17H18N3O4F2: 366.12655; found: 366.1269.
Example 34




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-chlorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (4-chlorophenyl)methanamine. Pale pink solid. 1H-NMR (500 MHz, CDCl3) δ ppm: 7.77 (1H, br), 7.33-7.35 (2H, m), 7.25-7.28 (2H, m), 4.59 (2H, d, J=6.4 Hz), 4.04 (4H, ddd, J=14.0, 7.9, 2.7 Hz), 1.58 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.12, 158.48, 151.87, 146.28, 135.72, 133.98, 129.92, 129.18, 129.09, 125.90, 75.92, 58.12, 43.45, 42.61, 28.06. HRMS [M+H]+ calcd for C17H19N3O4Cl: 364.10642; found: 364.1060.
Example 35




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2,4-difluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (2,4-difluorophenyl)methanamine. Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 7.86 (1H, t, J=5.6 Hz), 7.34-7.40 (1H, m), 6.83-6.90 (2H, m), 4.62 (2H, d, J=6.4 Hz), 4.01-4.06 (4H, m), 1.59 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.09, 158.43, 151.81, 146.22, 131.30, 131.25, 131.22, 131.18, 125.87, 120.37, 120.34, 111.85, 111.82, 111.68, 111.65, 104.47, 104.27, 104.07, 75.94, 58.12, 43.43, 37.00, 28.07. HRMS [M+H]+ calcd for C17H18N3O4F2: 366.12655; found: 366.1281.
Example 36




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-chloro-4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (2-chloro-4-fluorophenyl)methanamine. Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 8.05 (1H, br), 7.41 (1H, dd, J=8.4, 6.0 Hz), 7.17 (1H, dd, J=8.2, 2.4 Hz), 6.99 (1H, dd, J=8.2, 2.7 Hz), 4.66 (2H, d, J=6.4 Hz), 4.04 (4H, s), 1.60 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 167.88, 163.30, 161.31, 158.70, 151.77, 146.08, 134.55, 134.47, 131.65, 131.58, 130.70, 130.67, 126.12, 117.48, 117.28, 114.68, 114.51, 109.67, 75.99, 58.10, 43.51, 40.82, 28.08. HRMS [M+H]+ calcd for C17H18N3O4FCl: 382.09644; found: 382.0987.
Example 37




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2,4-dimethylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (2,4-dimethylphenyl)methanamine. Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 12.08 (1H, brs), 7.63 (1H, br), 7.15 (1H, d, J=7.6 Hz), 7.00-7.05 (2H, m), 4.57 (2H, d, J=5.8 Hz), 4.01 (4H, s), 2.32 (3H, s), 2.32 (3H, s), 1.55 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 167.97, 157.98, 151.74, 146.35, 137.95, 136.22, 131.90, 131.67, 128.38, 127.10, 125.64, 75.84, 58.21, 43.21, 41.15, 28.07, 21.11, 19.13. HRMS [M+H]+ calcd for C19H24N3O4: 358.17669; found: 358.1771.
Example 38




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,5-dimethylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (3,5-dimethylphenyl)methanamine. Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 12.09 (1H, s), 7.72-7.80 (1H, br), 6.95 (1H, s), 6.94 (2H, s), 4.55 (2H, d, J=6.4 Hz), 4.00-4.04 (4H, s), 2.32 (6H, s), 1.57 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.19, 157.89, 151.73, 146.44, 138.71, 137.25, 129.58, 125.61, 125.48, 75.86, 58.23, 43.18, 43.06, 28.08, 21.38. HRMS [M+H]+ calcd for C19H24N3O4: 358.17669; found: 358.1758.
Example 39




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-2-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (4-fluoro-2-methylphenyl)methanamine. Off white solid, 1H-NMR (500 MHz, CDCl3) δ ppm: 11.97 (1H, s), 7.64 (1H, br), 7.23 (1H, dd, J=8.2, 5.8 Hz), 6.87-6.94 (2H, m), 4.57 (2H, d, J=6.1 Hz), 4.02 (4H, s), 2.36 (3H, s), 1.56 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.07, 163.40, 161.44, 157.82, 151.86, 146.43, 138.81, 138.75, 130.78, 130.75, 129.97, 129.90, 125.42, 117.69, 117.52, 113.20, 113.03, 75.81, 58.23, 43.21, 40.71, 28.09, 19.32. HRMS [M+H]+ calcd for C18H21N3O4F: 362.15162; found: 362.1521.
Example 40




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-1-naphthalenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 37. White solid. 1H-NMR (500 MHz, CDCl3) δ ppm: 12.00 (1H, s), 8.15-8.20 (1H, m), 8.05 (1H, d, J=8.2 Hz), 7.73 (1H, br), 7.58-7.65 (2H, m), 7.43 (1H, dd, J=7.8, 5.3 Hz), 7.12 (1H, dd, J=10.1, 7.9 Hz), 5.02 (2H, d, J=6.1 Hz), 3.99 (4H, ddd, J=13.8, 8.0, 2.9 Hz), 1.49 (6H, s). 13C-NMR (126 MHz, CDCl3) δ ppm: 168.01, 160.13, 158.12, 157.77, 151.87, 146.51, 132.72, 132.68, 128.59, 128.56, 127.86, 126.54, 126.46, 125.42, 124.45, 123.23, 121.65, 121.60, 109.00, 108.84, 75.76, 58.21, 43.14, 40.86, 27.99. HRMS [M+H]+ calcd for C21H21N3O4F: 398.15162; found: 398.1536.
Example 41




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-2-methoxyphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (4-fluoro-2-methoxyphenyl)methanamine. HRMS [M+H]+ calcd for C18H2FN3O5: 378.1465; found: 378.1480. Light yellow crystals; 1H NMR (CDCl3, 500 MHz) δ ppm: 1.58 (6H, s, gem-di-Me), 3.88 (3H, s, OMe), 4.00 (4H, s, CH2), 4.53 (2H, d, J=6.5 Hz, CH2), 6.61-6.64 (2H, m, Ar—Hs), 7.24 (1H, m, Ar—Hs); 13C NMR (CDCl3, 125.8 Hz) δ ppm: 28.03 (CH3), 38.79 (CH2), 43.09 (CH2), 55.72 (CH3), 58.27 (CH2), 75.78 (C), 99.15, 99.35 (d, J=27 Hz, CH), 106.97, 107.14 (d, J=21 Hz, CH), 121.17, 121.20 (d, J=3.8 Hz, C), 125.75 (C), 130.44, 130.51 (d, J=9.6 Hz, CH), 146.26 (C), 151.50 (C), 157.87 (C═O), 158.77, 158.83 (d, J=9.6 Hz, C), 162.63, 164.48 (d, J=234 Hz, CF), 167.81 (C═O); HRMS (ESI) calcd for C18H21FN3O5 (M+H) 378.1465, found 378.1480; UV (MeOH) λmax 219 nm (ε1.66×104), 245 (ε9.69×103), 305 (ε7.70×103); Anal. Calcd for C18H20FN3O5.0.2H2O: C, 55.95; H, 5.48; N, 10.88; found C, 55.99; H, 5.11; N, 10.63.
Examples 42-43
Examples 42-43 can be prepared from intermediate 31, ethyl 9,9-diethyl-3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and the indicated amines according to the methods described for examples 1, 19 and 20.
Example 42




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9,9-diethyl-N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from 4-fluorobenzylamine. 1H NMR (500 MHz, CDCl3) δ ppm: 11.96 (1H, br), 7.76 (1H, br), 7.30 (2H, m), 7.06 (2H, m), 4.58 (2H, d, J=6.4 Hz), 4.00, (4H, m), 1.93 (2H, m), 1.86 (2H, m), 0.86 (6H, t, J=7.3 Hz). 13C NMR (500 MHz, CDCl3) δ ppm: 168.36, 163.46, 157.87, 151.61, 143.23, 133.14, 129.50, 125.58, 115.99, 115.82, 80.89, 58.46, 43.13, 42.50, 31.36, 7.79. HRMS [M+H]+ calcd for C19H23N3O4F: 376.16727; found: 376.1675.
Example 43




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9,9-diethyl-N-[(4-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-). The title compound can be prepared from 3-methyl, 4-fluorobenzylamine. 1H NMR (500 MHz, CDCl3) δ ppm: 11.99 (1H, br), 7.74 (1H, br), 7.15-7.09 (2H, m), 6.99 (1H, m), 4.54 (2H, d, J=6.1 Hz), 4.00, (4H, m), 2.27 (1H, s), 1.93 (2H, m), 1.86 (2H, m), 0.84 (6H, t, J=7.3 Hz). 13C NMR (126 MHz, CDCl3) δ ppm: 168.32, 161.97, 160.02, 157.84, 151.56, 146.23, 132.83, 130.96, 126.64, 125.59, 115.38, 80.88, 58.48, 43.11, 42.52, 31.36, 14.66, 7.79. HRMS [M+H]+ calcd for C20H25N3O4F: 390.18292; found: 390.1835.
Examples 44-45
Examples 44-45 can be prepared from intermediate 36, 3-hydroxy-10,10-dimethyl-4-oxo-6,7,8,10-tetrahydro-4H-pyrimido[2,1-c][1,4]oxazepine-2-carboxylate and the indicated amines according to the methods described for examples 1, 19 and 20.
Example 44




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. The title compound can be prepared from 4-fluorobenzylamine 1H NMR (500 MHz, CDCl3) δ ppm: 11.97 (1H, s), 7.72 (1H, br), 7.31 (1H, d, J=8.5 Hz), 7.30 (1H, d, J=8.5 Hz), 7.05 (1H, t, J=8.5 Hz), 4.58 (2H, d, J=6.4 Hz), 4.57 (2H, br), 3.67 (2H, t, J=6.4 Hz), 1.95 (2H, p, J=6.1 Hz), 1.57 (6H, s). 13C NMR (126 MHz, CDCl3) δ ppm: 168.32, 163.45, 161.49, 158.20, 153.63, 147.44, 133.17, 129.50, 129.43, 124.70, 115.97, 115.81, 82.29, 60.87, 42.47, 38.68, 27.82, 27.30. HRMS [M+H]+ calcd for C18H21N3O4F: 362.15162; found: 362.1530.
Example 45




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[(4-fluoro-3-methylphenyl)methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. The title compound can be prepared from 3-methyl, 4-fluorobenzylamine. 1H NMR (500 MHz, CDCl3) δ ppm: 12.00 (1H, s), 7.70 (1H, br), 7.14 (1H, m), 7.1 (1H, m), 6.98 (1H, t, J=8.9 Hz), 4.56 (2H, br), 4.54 (2H, d, J=6.4 Hz), 3.68 (2H, t, J=6.4 Hz), 2.27 (3H, s), 1.95 (2H, p, J=6.1 Hz), 1.57 (6H, s). 13C NMR (126 MHz, CDCl3) δ ppm: 168.26, 164.98, 160.02, 158.22, 153.59, 147.44, 132.82, 130.97, 126.58, 125.46, 124.75, 115.38, 82.30, 60.87, 42.51, 38.68, 27.83, 27.31, 14.66. HRMS [M+H]+ calcd for C19H23N3O4F: 376.16727; found: 376.1686.
Examples 46-51
Examples 46-52 can be prepared from the indicated intermediates according to the method provide for example 46.
Example 46




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[2-(4-morpholinyl)-2-oxoethoxy]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. A solution of intermediate 157, 3-benzyloxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydro-pyrimido[2,1-c][1,4]oxazine-2-carboxylic acid 4-fluoro-2-(2-morpholin-4-yl-2-oxo-ethoxy)-benzylamide, (187 mg, 0.32 mmol) in trifluoroacetic acid (2 mL) was stirred at room temperature for 2.5 hrs, after which the mixture was concentrated in vacuo to dryness. The residual oil was crystallized from 95% ethanol to provide 120 mg (0.25 mmol, Yield 77%) of the title compound as a white crystalline powder: 1H NMR (CDCl3, 500 MHz) δ ppm: 1.57 (6H, s, Me), 3.51, 3.64 (4H, brs, NCH2), 3.70 (4H, m, OCH2), 3.99 (4H, s, NCH2, OCH2), 4.60 (2H, d, J=6 Hz, NCH2), 4.76 (2H, s, OCH2), 6.59 (1H, dd, J=10, 2.5 Hz, Ar—H), 6.63 (1H, dt, J=2.5, 8 Hz, Ar—H), 7.29 (1H, dd, J=6.5, 8.5 Hz, Ar—H), 8.25 (1H, t, J=6 Hz, NH), 12.2 (br, OH). 13C NMR (CDCl3, 125.77 Hz) δ ppm: 27.93 (CH3), 38.44 (NCH2), 42.39 (NCH2), 43.14 (NCH2), 45.31 (NCH2), 58.19 (OCH2), 66.40 (OCH2), 66.59, 66.86 (OCH2), 75.94 (C), 100.26, 100.46 (d, J=26 Hz, CH), 108.19, 108.36 (d, J=21 Hz, CH), 122.03, 122.06 (d, J=3 Hz, C), 125.84 (C), 131.06, 131.14 (d, J=11 Hz, CH), 146.37 (C), 151.46 (C), 157.06, 157.14 (d, J=11 Hz, C), 157.96 (C═O), 162.25, 164.21 (d, J=248 Hz, CF), 165.47 (C═O), 168.23 (C═O); HRMS calcd for C23H28N4O7F(M+H) 491.1942, found 491.1958; UV (MeOH) λmax 249 nm (ε7.84×103), 290 nm (ε3.06×103), 303 nm (ε2.2×103); Anal. Calcd for C23H27N4O7F.1.7H2O: C, 53.01; H, 5.88; N, 10.75; found C, 52.53; H, 5.37; N, 10.48.
Example 47




Benzoic acid, 5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]-, methyl ester. The title compound can be prepared from intermediate 154, methyl 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorobenzoate. White solid; 1H NMR (300 MHz, CDCl3) δ ppm: 11.94 (1H, br s), 8.76 (1H, t, J=6.77 Hz), 7.69 (1H, dd, J=9.2, 2.9 Hz), 7.53 (1H, dd, J=8.4, 5.5 Hz), 7.15-7.22 (1H, m), 4.71 (2H, d, J=7.0 Hz), 3.97 (4H, s), 3.89-3.94 (3H, m), 1.56 (6H, s); HRMS (ESI) calcd for C19H20FN4O6 (M+H) 406.1414, found 406.1432.
Example 48




Benzoic acid, 5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]-. The title compound can be prepared from intermediate 190, 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorobenzoic acid. White solid; 1H NMR (500 MHz, CDCl3) δ ppm: 11.92 (1H, br s), 8.68 (1H, t, J=6.4 Hz), 7.80 (1H, dd, J=8.7, 2.6 Hz), 7.60 (1H, dd, J=8.5, 5.5 Hz), 7.30 (1H, dt, J=8.1, 2.8 Hz), 4.78 (2H, d, J=6.7 Hz), 4.00 (4H, s), 1.58 (6H, s); HRMS (ESI) calcd for C18H18FN3O6 (M+H) 392.1258, found 392.1250.
Example 49




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(methylamino)carbonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 143, N-(4-fluoro-2-(methylcarbamoyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (500 MHz, DMSO-d6) δ ppm: 11.97 (1H, br s), 9.46 (1H, br s), 8.55-8.51 (1H, m), 7.40-7.38 (1H, m), 7.32-7.27 (2H, m), 4.56 (2H, d, J=6.1 Hz), 3.97 (2H, t, J=4.9 Hz), 3.82 (2H, t, J=4.9 Hz), 2.80 (3H, d, J=4.6 Hz), 1.55 (6H, s). HRMS (M+H) calcd for C19H22FN4O5: 405.1574; found: 405.1588.
Example 50




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(cyclopropylamino)carbonyl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 155, N-(2-(cyclopropylcarbamoyl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. White solid; 1H NMR (300 MHz, CDCl3) δ ppm: 11.98 (1H, br s), 8.84 (1H, t, J=7.32 Hz), 7.48 (1H, dd, J=9.0, 5.3 Hz), 7.05-7.16 (2H, m), 6.20-6.31 (1H, br s), 4.56 (2H, d, J=6.6 Hz), 3.92-4.02 (4H, m), 2.90 (1H, dt, J=7.1, 3.3 Hz), 1.59 (6H, s), 0.88 (2H, q, J=6.6 Hz), 0.57-0.66 (2H, m); HRMS (ESI) calcd for C21H23FN4O5 (M+H) 431.1731, found 431.1734.
Example 51




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[[(2-hydroxyethyl)amino]carbonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 191, N-(2-((2-aminoethyl)carbamoyl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (500 MHz, CDCl3) δ ppm: 11.98 (1H, br s), 8.88 (1H, t, J=6.0 Hz), 7.51 (1H, dd, J=8.2, 5.5 Hz), 7.21 (1H, dd, J=8.5, 2.8 Hz), 7.14 (1H, dt, J=8.3, 2.6 Hz), 6.57-6.63 (1H, m), 4.58 (2H, d, J=6.7 Hz), 4.00 (4H, s), 3.87 (2H, t, J=5.1 Hz), 3.63-3.68 (2H, m), 1.60 (6H, s); HRMS (ESI) calcd for C20H23FN4O6 (M+H) 435.1680, found 435.1700.
Example 52




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(4-morpholinylcarbonyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 156, N-(4-fluoro-2-(morpholine-4-carbonyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. White solid; 1H NMR (500 MHz, CDCl3) δ ppm: 11.94 (1H, s), 8.46 (1H, t, J=5.5 Hz), 7.47 (1H, dd, J=8.5, 5.5 Hz), 7.10 (1H, dt, J=8.4, 2.4 Hz), 6.94 (1H, dd, J=8.2, 2.4 Hz), 4.00 (4H, s), 3.79-3.88 (2H, br), 3.78 (2H, br), 3.61 (2H, br), 3.31-3.40 (2H, br), 1.61 (6H, s); HRMS (ESI) calcd for C22H25FN4O6 (M+H) 461.1836, found 461.1852.
Examples 53-60
Examples 53-60 can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and the indicated amines according to the method described for the synthesis of examples 1, 19 and 20.
Example 53




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1H-imidazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 89. 1H-NMR (500 MHz, CDCl3) δ ppm: 11.66 (1H, bs), 8.07 (1H, s), 7.74 (1H, t, J=5.5 Hz), 7.57 (1H, dd, J=8.7, 5.9 Hz), 7.35 (1H, s), 7.25-7.21 (2H, m), 7.08 (1H, dd, J=8.2, 2.4 Hz), 4.42 (2H, d, J=6.4 Hz), 4.01 (4H, s), 1.59 (6H, s). HRMS [M+H]+ calcd for C20H21N5O4F: 414.15777; found: 414.1563. Anal calcd for C20H20N5O4F.0.25H2O: C, 57.48; H, 4.94; N, 16.76; F, 4.55; found: C, 57.77; H, 4.89; N, 16.29; F, 4.48.
Example 54




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[5-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-e. The title compound can be prepared from intermediate 97. 1H NMR (500 MHz, CDCl3) δ ppm: 11.82 (1H, s), 8.70 (1H, t, J=6.5 Hz), 8.39 (1H, s), 8.17 (1H, s), 7.40 (1H, dd, J=8.6, 2.7 Hz), 7.34 (1H, dd, J=8.9, 4.9 Hz), 7.17-7.13 (1H, m), 4.44 (2H, d, J=6.7 Hz), 4.01 (4H, s), 1.62 (6H, s). HRMS (M+H) calcd for C19H20FN6O4: 415.1530; found: 415.1544. Anal calcd for C19H19FN6O4: C, 55.07; H, 4.62; N, 20.28; F, 4.58; found: C, 54.83; H, 4.51; N, 19.89; F, 4.56.
Example 55




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[3-fluoro-2-(tetrahydro-1,1-dioxido-2H-1,2-thiazin-2-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 93. White solid. 36% yield. 1H-NMR (500 MHz, CDCl3) δ ppm: 11.96 (1H, s), 8.42-8.39 (2H, m), 7.14-7.11 (1H, m), 4.97 (1H, dd J=14.3, 8.8 Hz), 4.37 (1H, dd, J=14.3, 4.0 Hz), 4.00 (4H, s), 3.86-3.80 (1H, m), 3.75-3.70 (1H, m), 3.34-3.24 (2H, m), 2.44-2.39 (2H, m), 2.08-2.00 (1H, m), 1.83-1.77 (1H, m), 1.60 (3H, s), 1.57 (3H, s). HRMS [M+H]+ calcd for C21H26N4O6FS: 481.15572; found: 481.1559. Anal calcd for C21H25N4O6FS: C, 52.49; H, 5.24; N, 11.66; S, 6.67; F, 3.95; found: C, 52.43; H, 5.21; N, 11.61; S, 6.56; F, 4.16.
Example 56




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[3-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 95. White solid. 1H-NMR (500 MHz, CDCl3) δ ppm: 11.86 (1H, s), 8.76 (1H, brs), 8.46 (1H, d, J=3.0 Hz), 8.22 (1H, s), 7.48-7.47 (2H, m), 7.29-7.26 (1H, m), 4.44 (2H, d, J=6.7 Hz), 4.01 (4H, s), 1.63 (6H, s). HRMS [M+H]+ calcd for C19H20N6O4F: 415.15302; found: 415.1541. Anal calcd for C19H19N6O4F: C, 55.07; H, 4.62; N, 20.28; F, 4.58; found: C, 55.18; H, 4.42; N, 20.17; F, 4.51.
Example 57




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(2H-1,2,3-triazol-2-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 77. Pale orange solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 11.99 (1H, s), 8.99 (1H, t, J=6.4 Hz), 7.91 (2H, s), 7.65-7.58 (2H, m), 7.10 (1H, td, J=8.1, 2.6 Hz), 4.61 (2H, d, J=7.0 Hz), 3.97 (4H, s), 1.55 (6H, s). HRMS [M−H]− calcd for C19H18N6O4F: 413.13736; found: 413.1354. Anal calcd for C19H17N6O4F: C, 55.07; H, 4.62; N, 20.28; F, 4.58; found: C, 54.94; H, 4.78; N, 20.32; F, 4.53.
Example 58




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-fluoro-4-(2H-1,2,3-triazol-2-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 77. Pale brown solid 1H-NMR (300 MHz, CDCl3) δ ppm: 11.85 (1H, s), 7.88-7.82 (3H, m), 7.79 (2H, s), 7.47 (1H, t, J=8.3 Hz), 4.67 (2H, d, J=6.2 Hz), 3.99 (4H, s), 1.56 (6H, s). HRMS [M+H]+ calcd for C19H20N6O4F: 415.15302; found: 415.1513. Anal calcd for C19H19N6O4F: C, 55.07; H, 4.62; N, 20.28; F, 4.58; found: C, 54.94; H, 4.76; N, 19.94; F, 4.26.
Example 59




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-bromo-4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (2-bromo-4-fluorophenyl)methanamine. White needles. 1H-NMR (300 MHz, CDCl3) δ ppm: 11.78 (1H, s), 8.08 (1H, t, J=6.0 Hz), 7.39 (1H, dd, J=8.8, 5.8 Hz), 7.31 (1H, dd, J=8.0, 2.6 Hz), 7.01 (1H, dt, J=8.2, 2.6 Hz), 4.61 (2H, d, J=6.6 Hz), 3.99 (4H, s), 1.56 (6H, s). HRMS [M+H]+ calcd for C17H18N3O4FBr: 426.04648; found: 426.0465.
Example 60




1H-1,2,4-Triazole-3-carboxylic acid, 1-[5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl], methyl este. The title compound can be prepared from intermediate 91. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 11.90 (1H, s), 8.49 (1H, s), 8.36 (1H, t, J=6.2 Hz), 7.72 (1H, dd, J=8.8, 5.9 Hz), 7.26-7.20 (1H, m), 7.14 (1H, dd, J=8.4, 2.6 Hz), 4.49 (2H, d, J=6.6 Hz), 4.01 (3H, s), 3.98 (4H, s), 1.58 (6H, s). HRMS [M+H]+ calcd for C21H22N6O6F: 473.1585; found: 473.1563. Anal calcd for C21H21N6O6F.0.5H2O: C, 52.39; H, 4.61; N, 17.46; F, 3.95; found: C, 52.14; H, 4.70; N, 17.41; F, 4.12.
Example 61




1H-1,2,4-Triazole-3-carboxylic acid, 1-[5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-. To a solution of example 60, 1H-1,2,4-triazole-3-carboxylic acid, 1-[5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, methyl ester, (2.156 g, 4.6 mmol) in tetrahydrofuran (200 mL) and water (50 mL) at 0° C. was added lithium hydroxide monohydrate (0.58 g, 13.8 mmol). The mixture was stirred at 0° C. 2 h and at room temp for 1 h. The resulting solution was partitioned between ethyl acetate and water. The aqueous phase was acidified with 1 N HCl and extracted with ethyl acetate and CH2Cl2. The organic extracts were combined, dried (Na2SO4) and concentrated to give the title compound as a white solid (2.05 g, 97% yield). 1H-NMR (300 MHz, CDCl3) δ ppm: 11.91 (1H, bs), 8.55 (1H, s), 8.43 (1H, t, J=6.6 Hz), 7.77 (1H, dd, J=8.8, 5.9 Hz), 7.28-7.23 (1H, m), 7.16 (1H, dd, J=8.0, 2.6 Hz), 4.47 (2H, d, J=6.9 Hz), 3.98 (4H, s), 1.59 (6H, s). HRMS [M+H]+ calcd for C20H20N6O6F: 459.14285; found: 459.1442.
Examples 62-72
Examples 62-72 can be prepared from intermediate 61, 1H-1,2,4-triazole-3-carboxylic acid, 1-[5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, according to the method described for the synthesis of example 62.
Example 62




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-(4-morpholinylcarbonyl)-1H-1,2,4-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. To a solution of example 61, 1H-1,2,4-triazole-3-carboxylic acid, 1-[5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, (0.0259 g, 0.057 mmol) in DMF (2 mL), at 0° C., was added O-(7-azabenzotriazol-1-yl)-N,N,N1,N1-tetramethyluronium hexafluorophosphate (0.044 g, 0.115 mmol). The solution was stirred at 0° C. for 10 min before adding morpholine (0.025 mL, 0.285 mmol) after which it was stirred at room temp for 2 h. Purification by reverse phase preparative HPLC chromatography (YMC Combiprep ODS-A, 30 mm×50 mm, MeOH/H2O/0.1% CF3CO2H) gave the title compound as a white solid (0.015 g, 50% yield). 1H-NMR (500 MHz, CDCl3) δ ppm: 8.50 (1H, t, J=6.9 Hz), 8.45 (1H, s), 7.69 (1H, dd, J=8.7, 5.9 Hz), 7.22 (1H, dd, J=8.2, 2.3 Hz), 7.13 (1H, dd, J=8.2, 2.4 Hz), 4.49 (2H, s), 3.98 (4H, s), 3.88-3.72 (8H, m), 1.57 (6H, s). HRMS [M+H]+ calcd for C24H27N7O6F: 528.20069; found: 528.2025.
Example 63




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[3-[(dimethylamino)carbonyl]-1H-1,2,4-triazol-1-yl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. Pale purple solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.51-8.45 (2H, m), 7.70 (1H, dd, J=8.4, 5.9 Hz), 7.24-7.18 (1H, m), 7.12 (1H, dd, J=8.2, 2.4 Hz), 4.47 (2H, d, J=6.9 Hz), 3.97 (4H, s), 3.24 (3H, s), 3.15 (3H, s), 1.54 (6H, s). HRMS [M+H]+ calcd for C22H25N7O5F: 486.19013; found: 486.1887.
Example 64




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-[[(methylsulfonyl)amino]carbonyl]-1H-1,2,4-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.52 (1H, s), 8.36 (1H, t, J=6.8 Hz), 7.70 (1H, dd, J=8.6, 5.7 Hz), 7.26-7.19 (1H, m), 7.13 (1H, dd, J=8.4, 2.6 Hz), 4.46 (2H, d, J=7.0 Hz), 3.95 (4H, s), 3.39 (4H, s), 1.57 (6H, s). HRMS [M+H]+ calcd for C21H23N7O7FS: 536.1364; found: 536.1376. Anal calcd for C21H22N7O7FS.0.07CF3CO2H: C, 46.72; H, 4.09; N, 18.04; F, 4.23; found: C, 46.42; H, 3.91; N, 17.70; F, 4.17.
Example 65




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4, 6,7,9-tetrahydro-3-hydroxy-N-[(1R)-2-hydroxy-1-phenylethyl]-9,9-dimethyl-4-oxo-. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.43 (1H, s), 8.20 (1H, t, J=6.0 Hz), 7.64 (1H, dd, J=8.8, 5.9 Hz), 7.42-7.40 (1H, m), 7.26-7.19 (1H, m), 7.11 (1H, dd, J=8.0, 2.6 Hz), 4.53 (2H, d, J=6.6 Hz), 3.99 (4H, s), 3.06 (3H, d, J=4.7 Hz), 1.56 (6H, s). HRMS [M+H]+ calcd for C21H23N7O5F: 472.1745; found: 472.1741. Anal calcd for C21H22N7O5F: C, 53.50; H, 4.70; N, 20.79; F, 4.03; found: C, 53.22; H, 4.51; N, 20.70; F, 4.01.
Example 66




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[3-[(4-acetyl-1-piperazinyl)carbonyl]-1H-1,2,4-triazol-1-yl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.46 (1H, s), 8.36 (1H, t, J=5.7 Hz), 7.68 (1H, dd, J=8.6, 5.7 Hz), 7.27-7.21 (1H, m), 7.14 (1H, dd, J=8.2, 2.7 Hz), 4.48 (2H, d, J=6.6 Hz), 3.98 (4H, s), 3.94-3.61 (8H, m), 2.13 (3H, s), 1.55 (6H, s). HRMS [M+H]+ calcd for C26H30N8O6F: 569.2272; found: 569.2269. Anal calcd for C26H29N8O6F.0.8H2O: C, 53.57; H, 5.29; N, 19.22; F, 3.26; found: C, 53.48; H, 4.95; N, 19.21; F, 3.21.
Example 67




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-[[(2-hydroxyethyl)methylamino]carbonyl]-1H-1,2,4-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.54 (1H, t, J=7.3 Hz), 8.44 (1H, s), 7.66 (1H, dd, J=8.8, 5.8 Hz), 7.21-7.09 (2H, m), 4.44 (2H, d, J=4.4 Hz), 3.93 (4H, s), 3.86-3.78 (2H, m), 3.70-3.65 (2H, m), 3.13 (3H, s), 1.94 (1H, bs), 1.55 (6H, s). HRMS [M+H]+ calcd for C23H27N7O6F: 516.2007; found: 516.2011. Anal calcd for C23H26N7O6F: C, 53.59; H, 5.08; N, 19.02; F, 3.68; found: C, 53.31; H, 5.06; N, 18.80; F, 3.60.
Example 68




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-[[[(4-fluorophenyl)sulfonyl]amino]carbonyl]-1H-1,2,4-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.47 (1H, s), 8.18-8.13 (2H, m), 7.68 (1H, dd, J=8.8, 5.9 Hz), 7.24-7.16 (3H, m), 7.08 (1H, dd, J=8.0, 2.6 Hz), 4.39 (2H, s), 3.96 (4H, s), 1.55 (6H, s). HRMS [M+H]+ calcd for C26H24N7O7F2S: 616.1426; found: 616.1426. Anal calcd for C26H23N7O7F2S: C, 50.73; H, 3.76; N, 15.92; F, 6.17; found: C, 50.49; H, 3.66; N, 15.98; F, 6.12.
Example 69




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-[(4-methyl-1-piperazinyl)carbonyl]-1H-1,2,4-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. Pale brown foam. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.43 (1H, s), 8.29 (1H, t, J=6.8 Hz), 7.63 (1H, dd, J=8.4, 5.9 Hz), 7.25-7.19 (1H, m), 7.11 (1H, dd, J=8.0, 2.6 Hz), 4.46 (2H, d, J=6.6 Hz), 3.95 (4H, s), 3.30 (4H, bs), 2.86 (3H, s), 1.91 (4H, bs), 1.53 (6H, s). HRMS [M+H]+ calcd for C25H30N8O5F: 541.2323; found: 541.2341. Anal calcd for C25H29N8O5Fe0.5CF3CO2H.0.5H2O: C, 46.67; H, 4.41; N, 15.55; F, 14.50; found: C, 46.86; H, 4.44; N, 15.67; F, 14.48.
Example 70




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[3-[[[2-(dimethylamino)ethyl]amino]carbonyl]-1H-1,2,4-triazol-1-yl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. Pale brown foam. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.39 (1H, s), 7.58 (1H, dd, J=8.6, 5.7 Hz), 7.23-7.14 (1H, m), 7.09 (1H, dd, J=8.4, 2.6 Hz), 4.50 (2H, s), 3.95 (4H, s), 3.84 (2H, t, J=5.5 Hz), 3.34 (2H, t, J=6.0 Hz), 2.89 (6H, s), 1.55 (6H, s). HRMS [M+H]+ calcd for C24H30N8O5F: 529.2323; found: 529.2315.
Example 71




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[3-[[[2-(dimethylamino)ethyl]methylamino]carbonyl]-1H-1,2,4-triazol-1-yl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. Pale brown foam. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.44 (1H, s), 8.21 (1H, t, J=6.5 Hz), 7.67-7.58 (1H, m), 7.24-7.09 (2H, m), 4.48-4.45 (2H, m), 3.95 (4H, s), 3.94-3.89 (2H, m), 3.37-3.33 (2H, m), 2.91 (6H, s), 2.88 (3H, s), 1.56 (6H, s). HRMS [M+H]+ calcd for C25H32N8O5F: 543.2480; found: 543.2491. Anal calcd for C25H31N8O5F.CF3CO2H: C, 45.20; H, 4.32; N, 14.54; F, 17.26; found: C, 45.13; H, 4.14; N, 14.74; F, 17.01.
Example 72




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-[[(2-hydroxyethyl)amino]carbonyl]-1H-1,2,4-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 8.41 (1H, s), 8.32 (1H, t, J=6.2 Hz), 7.63 (1H, dd, J=8.4, 5.9 Hz), 7.20 (1H, dt, J=8.4, 2.6 Hz), 7.11 (1H, dd, J=8.4, 2.6 Hz), 4.52-4.50 (2H, m), 3.95 (4H, s), 3.78 (2H, t, J=5.1 Hz), 3.59 (2H, t, J=5.1 Hz), 1.56 (6H, s). HRMS [M+H]+ calcd for C22H25N7O6F: 502.1850; found: 502.1850.
Example 73




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-2-iodophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 174, N-(4-fluoro-2-iodobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.63 (6H, s, 2×CH3), 4.05 (4H, s, 2×CH2), 4.63 (2H, d, J=7.1 Hz, NCH2), 7.11 (1H, m, aromatic), 7.42 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 7.62 (1H, dd, J=2.5 Hz and J=8.1 Hz, aromatic), 8.20 (1H, broad t, NH), 11.82 (1H, s, OH). HRMS (ESI+) calculated for C17H18FIN3O4 [M+H+]: 474.0326; found: 474.0328.
Examples 74-77
Examples 74-77 can be prepared from the indicated intermediates by hydrogenolysis or trifluoroacetic acid mediated hydrolysis.
Example 74




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(5-fluoro[1,1′-biphenyl]-2-yl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 176, N-(4-fluoro-2-phenyl-benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.56 (6H, s, 2×CH3), 4.03 (4H, s, 2×CH2), 4.56 (2H, d, J=6.0 Hz, NCH2), 7.03 (1H, dd, J=2.5 Hz and J=9.3 Hz, aromatic), 7.09 (1H, m, aromatic), 7.36 (2H, m, aromatics), 7.42-7.51 (5H, m, aromatics and NH), 11.96 (1H, s, OH). HRMS (ESI+) calculated for C23H23FN3O4 [M+H+]: 424.1673; found: 424.1675.
Example 75




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(3-pyridinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1HNMR 400 MHz (CDCl3) δ ppm: 1.57 (6H, s, 2×CH3), 4.04 (4H, s, 2×CH2), 4.55 (2H, d, J=6.1 Hz, NCH2), 7.04 (1H, dd, J=2.5 Hz and J=9.1 Hz, aromatic), 7.16 (1H, m, aromatic), 7.43 (1H, m, aromatic), 7.49 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 7.52 (1H, broad t, NH), 7.71 (1H, m, aromatic), 8.63 (1H, m, aromatic), 8.70 (1H, m, aromatic), 11.84 (1H, s, OH). HRMS (ESI+) calculated for C22H22FN4O4 [M+H+]: 425.1625; found: 425.1616.
Example 76




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(2-methoxy-3-pyridinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. Hydrogenolysis of intermediate 175, N-(4-fluoro-2-(2-methoxypyridin-3-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide gave the title material as a white solid; mp 227° C. 1HNMR 400 MHz (CDCl3) δ ppm: 1.58 (6H, s, 2×CH3), 3.99 (3H, s, OCH3), 4.04 (4H, s, 2×CH2), 4.43 (2H, broad, NCH2), 6.97 (1H, dd, J=2.5 Hz and J=8.5 Hz, aromatic), 7.03 (1H, dd, J=5.0 Hz and J=8.5 Hz, aromatic), 7.12 (1H, m, aromatic), 7.45 (1H, dd, J=4.5 Hz and J=8.6 Hz, aromatic), 7.53 (1H, dd, J=2.0 Hz and J=7.1 Hz, aromatic), 7.57 (1H, broad t, NH), 8.28 (1H, dd, J=2.5 Hz and J=5.0 Hz, aromatic), 12.03 (1H, s, OH). HRMS (ESI+) calculated for C23H24FN4O5 [M+H+]: 455.1731; found: 455.1737.
Example 77




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(6-methoxy-3-pyridinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo. 1HNMR 400 MHz (CDCl3) δ ppm: 1.58 (6H, s, 2×CH3), 4.02 (3H, s, OCH3), 4.04 (4H, s, 2×CH2), 4.55 (2H, d, J=6.1 Hz, NCH2), 6.87 (1H, d, J=9.0 Hz, aromatic), 7.01 (1H, dd, J=2.0 Hz and J=9.0 Hz, aromatic), 7.12 (1H, m, aromatic), 7.46 (1H, dd, J=5.5 Hz and J=8.6 Hz, aromatic), 7.55 (1H, broad t, NH), 7.60 (1H, dd, J=2.0 Hz and J=8.6 Hz, aromatic), 8.17 (1H, d, J=2.0 Hz, aromatic), 11.89 (1H, s, OH). HRMS (ESI+) calculated for C23H24FN4O5 [M+H+]: 455.1731; found: 455.1717.
Examples 78-80
Examples 78-80 can be prepared from the indicated intermediates according to the method described for example 78.
Example 78




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(1,2-dihydro-2-oxo-3-pyridinyl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. A solution of intermediate 175, N-(4-fluoro-2-(2-methoxypyridin-3-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, (0.125 g, 0.23 mmol) in acetonitrile (5 ml) was treated with sodium iodide (0.090 g, 0.6 mmol) and chlorotrimethylsilane (0.45 ml, 3.5 mmol), sealed in a pressure resistant vessel and heated at 80° C. for 1.5 h. The mixture was diluted with ethyl acetate washed with water and brine then dried over anhydrous magnesium sulfate. Filtration and removal of solvent provided the title compound. 1HNMR 400 MHz (CDCl3) δ ppm: 1.60 (6H, s, 2×CH3), 4.03 (4H, s, 2×CH2), 4.5 (2H, broad, NCH2), 6.47 (1H, m, aromatic), 6.95 (1H, dd, J=2 Hz and J=9 Hz, aromatic), 7.12 (1H, m, aromatic), 7.45 (1H, dd, J=2 Hz and J=7 Hz, aromatic), 7.50 (1H, dd, J=6 Hz and J=9 Hz, aromatic), 7.53 (1H, dd, J=2 Hz and J=7 Hz, aromatic), 8.64 (1H, broad t, NH). HRMS (ESI+) calculated for C22H22FN4O5 [M+H+]: 441.1574; found: 441.1585.
Example 79




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(1,6-dihydro-6-oxo-3-pyridinyl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dim ethyl-4-oxo-. The title compound can be prepared from example 77. 1HNMR 400 MHz (CDCl3) δ ppm: 1.59 (6H, s, 2×CH3), 4.02 (3H, s, OCH3), 4.05 (4H, s, 2×CH2), 4.55 (2H, d, J=6.6 Hz, NCH2), 6.95 (1H, d, J=9.0 Hz, aromatic), 7.00 (1H, dd, J=2.6 Hz and J=9.0 Hz, aromatic), 7.17 (1H, m, aromatic), 7.46 (1H, dd, J=5.5 Hz and J=8.6 Hz, aromatic), 7.63 (1H, d, J=2.6 Hz, aromatic), 7.71 (1H, broad t, NH), 7.79 (1H, dd, J=2.6 Hz and J=9.1 Hz, aromatic). HRMS (ESI+) calculated for C22H22FN4O5 [M+H+]: 441.1574; found: 441.1570.
Example 80




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(1,6-dihydro-6-oxo-2-pyridinyl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from N-(4-fluoro-2-(6-methoxypyridin-2-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, which can be prepared according to the methods described for the synthesis of intermediate 175. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.62 (6H, s, 2×CH3), 4.04 (4H, s, 2×CH2), 4.60 (2H, d, J=7.0 Hz, NCH2), 6.38 (1H, d, J=7.0 Hz, aromatic), 6.68 (1H, d, J=9.0 Hz, aromatic), 7.14 (1H, dd, J=2.5 Hz and J=9.1 Hz, aromatic), 7.23 (1H, m, aromatic), 7.50 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 7.58 (1H, dd, J=7.0 Hz and J=9.0 Hz, aromatic), 8.15 (1H, broad t, NH).
Examples 81-93
Examples 81-93 can be prepared from the indicated intermediates by hydrogenolysis or trifluoroacetic acid mediated hydrolysis.
Example 81




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 152, N-(4-fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide 1HNMR 400 MHz (CDCl3) δ ppm: 1.64 (6H, s, 2×CH3), 2.59 (3H, s, CH3), 4.04 (4H, s, 2×CH2), 4.50 (2H, d, J=7.1 Hz, NCH2), 7.1 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.20 (1H, m, aromatic), 7.72 (1H, dd, J=6.0 Hz and J=8.6 Hz, aromatic), 8.34 (1H, s, CH), 8.80 (1H, broad t, NH), 12.11 (1H, s, OH). HRMS (ESI+) calculated for C20H22FN6O4 [M+H+]: 429.1687; found: 429.1675.
Example 82




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(5-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1HNMR 400 MHz (CDCl3) δ ppm: 1.66 (6H, s, 2×CH3), 2.50 (3H, s, CH3), 4.04 (4H, s, 2×CH2), 4.32 (2H, d, J=7.0 Hz, NCH2), 7.05 (1H, dd, J=2.5 Hz and J=8.1 Hz, aromatic), 7.27 (1H, m, aromatic), 7.70 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 8.04 (1H, s, CH), 8.61 (1H, broad t, NH), 11.90 (1H, s, OH). HRMS (ESI+) calculated for C20H22FN6O4 [M+H+]: 429.1687; found: 429.1688.
Example 83




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-fluoro-4-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 153, N-(2-fluoro-4-(3-methyl-1H-1,2,4-triazol-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.61 (6H, s, 2×CH3), 2.51 (3H, s, CH3), 4.05 (4H, s, 2×CH2), 4.72 (2H, d, J=6.6 Hz, NCH2), 7.44-7.55 (3H, m, aromatics), 7.94 (1H, broad t, NH), 8.46 (1H, s, CH), 11.86 (1H, s, OH). HRMS (ESI+) calculated for C20H22FN6O4 [M+H+]: 429.1687; found: 429.1695.
Example 84




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1,2,3-thiadiazol-4-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 172, N-(4-fluoro-2-(1,2,3-thiadiazol-4-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (DMSO-d6) δ ppm: 1.54 (6H, s, 2×CH3), 3.83 (2H, broad t, CH2), 3.96 (2H, broad t, CH2), 4.61 (2H, d, J=6.7 Hz, NCH2), 7.39 (1H, m, aromatic), 7.55 (1H, m, aromatic), 7.62 (1H, m, aromatic), 9.41 (1H, broad t, NH), 9.63 (1H, s, CH), 12.0 (1H, s, OH). HRMS (ESI+) calculated for C19H19FN5O4S [M+H+]: 432.1142; found: 432.1124.
Example 85




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1H-pyrazol-5-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 177, N-(4-fluoro-2-(1H-pyrazol-5-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.59 (6H, s, 2×CH3), 4.03 (4H, s, 2×CH2), 4.67 (2H, d, J=6.6 Hz, NCH2), 6.65 (1H, d, J=2.5 Hz, CH), 7.07 (1H, m, aromatic), 7.31 (1H, dd, J=2.5 Hz and J=9.8 Hz, aromatic), 7.56 (1H, dd, J=5.8 Hz and J=8.3 Hz, aromatic), 7.75 (1H, d, J=2.5 Hz, CH), 9.22 (1H, broad t, NH), 10.33 (1H, broad, NH), 12.2 (1H, s, OH). HRMS (ESI+) calculated for C20H21FN5O4 [M+H+]: 414.1578; found: 414.1560.
Example 86




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(5-methyl-2-oxazolyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 173, N-(4-fluoro-2-(5-methyloxazol-2-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.58 (6H, s, 2×CH3), 2.48 (3H, s, CH3), 4.01 (4H, s, 2×CH2), 4.77 (2H, d, J=7.0 Hz, NCH2), 6.96 (1H, s, CH), 7.13 (1H, m, aromatic), 7.61 (1H, dd, J=5.8 Hz and J=8.3 Hz, aromatic), 7.70 (1H, dd, J=3.2 Hz and J=9.6 Hz, aromatic), 9.76 (1H, broad t, NH), 12.15 (1H, s, OH). HRMS (ESI+) calculated for C21H22FN4O5 [M+H+]: 429.1574; found: 429.1564.
Example 87




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(ethylamino)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 170, N-(2-(ethylamino)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.30 (3H, t, J=7.3 Hz, CH3), 1.59 (6H, s, 2×CH3), 3.12 (2H, m, CH2), 4.04 (4H, s, 2×CH2), 4.52 (2H, d, J=6.6 Hz, NCH2), 5.09 (1H, broad, NH), 6.3-6.37 (2H, m, aromatics), 7.10 (1H, dd, J=6.6 Hz and J=8.1 Hz, aromatic), 7.67 (1H, broad t, NH), 11.93 (1H, s, OH). HRMS (ESI+) calculated for C19H24FN4O4 [M+H+]: 391.1782; found: 391.1774.
Example 88




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-ethynyl-4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 179, N-(2-ethynyl-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.61 (6H, s, 2×CH3), 3.46 (1H, s, CH), 4.04 (4H, s, 2×CH2), 4.73 (2H, d, J=6.5 Hz, NCH2), 7.1 (1H, m, aromatic), 7.26 (1H, dd, J=2.5 Hz and J=8.5 Hz, aromatic), 7.40 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 8.18 (1H, broad t, NH), 11.92 (1H, s, OH). HRMS (ESI+) calculated for C19H19FN3O4 [M+H+]: 372.1360; found: 372.1345.
Example 89




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(3-hydroxy-1-propynyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 182, N-(4-fluoro-2-(3-hydroxyprop-1-ynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.60 (6H, s, 2×CH3), 4.04 (4H, s, 2×CH2), 4.55 (2H, broad d, CH2), 4.73 (2H, d, J=6.6 Hz, NCH2), 7.07 (1H, m, aromatic), 7.20 (1H, dd, J=2.5 Hz and J=9.1 Hz, aromatic), 7.38 (1H, dd, J=5.3 Hz and J=8.3 Hz, aromatic), 7.95 (1H, broad t, NH), 11.90 (1H, s, OH). HRMS (ESI+) calculated for C20H21FN3O5 [M+H+]: 402.1465; found: 402.1463.
Example 90




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-[(methylsulfonyl)oxy]-1-propynyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 183, 3-[2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl]prop-2-ynyl methanesulfonate. 1HNMR 400 MHz (CDCl3) δ ppm: 1.62 (6H, s, 2×CH3), 3.16 (3H, s, CH3), 4.05 (4H, s, 2×CH2), 4.72 (2H, d, J=6.0 Hz, NCH2), 5.12 (2H, s, OCH2), 7.12 (1H, m, aromatic), 7.21 (1H, dd, J=2.6 Hz and J=8.6 Hz, aromatic), 7.45 (1H, dd, J=5.1 Hz and J=8.6 Hz, aromatic), 8.03 (1H, broad t, NH).
Example 91




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[3-(dimethylamino)-1-propynyl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 184, N-(2-(3-(dimethylamino)prop-1-ynyl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound was isolated as a trifluoroacetic acid salt. 1HNMR 400 MHz (CDCl3) δ ppm: 1.62 (6H, s, 2×CH3), 3.03 (6H, s, 2×CH3), 4.06 (4H, s, 2×CH2), 4.23 (2H, s, NCH2), 4.75 (2H, d, J=6.0 Hz, NCH2), 7.16 (1H, m, aromatic), 7.24 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.42 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 8.04 (1H, broad t, NH). HRMS (ESI+) calculated for C22H26FN4O4 [M+H+]: 429.1938; found: 429.1917.
Example 92




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-(methylsulfonyl)-1-propynyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 186, N-(4-fluoro-2-(3-(methylsulfonyl)prop-1-ynyl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.63 (6H, s, 2×CH3), 3.16 (3H, s, SCH3), 4.04 (4H, s, 2×CH2), 4.17 (2H, s, SCH2), 4.70 (2H, d, J=6.0 Hz, NCH2), 7.12 (1H, m, aromatic), 7.21 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.49 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 8.16 (1H, broad t, NH), 11.99 (1H, s, OH). HRMS (ESI+) calculated for C21H23FN3O6S [M+H+]: 464.1292; found: 464.1271.
Example 93




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[3-(tetrahydro-1,1-dioxido-2H-1,2-thiazin-2-yl)-1-propynyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1HNMR 400 MHz (CDCl3) δ ppm: 1.63 (6H, s, 2×CH3), 1.86 (2H, m, CH2), 2.28 (2H, m, CH2), 3.11 (2H, m, CH2), 3.53 (2H, m, CH2), 4.05 (4H, s, 2×CH2), 4.27 (2H, s, NCH2), 4.73 (2H, d, J=6.0 Hz, NCH2), 7.07 (1H, m, aromatic), 7.17 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.47 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 8.12 (1H, broad t, NH).
Example 94




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-[2-(phenylsulfonyl)ethyl]-. The title compound can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and 7-(aminomethyl)indolin-2-one according to the methods described for examples 1, 19 and 20. 1H NMR (500 MHz, DMSO-d6) δ ppm: 1.58 (s, 6), 3.50 (s, 2), 3.83 (m, 2), 3.97 (m, 2), 4.42 (d, 2), 6.9-7.13 (overlapping m, 3). Anal calcd C19H20N4O5: C, 59.36; H, 5.24; N, 14.57. Found: C, 59.61; H, 5.43; N, 14.46.
Example 95




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-2-hydroxyphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. A solution of intermediate 144, N-(4-fluoro-2-hydroxybenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, (0.070 g, 0.154 mmol) in dichloromethane (3 mL) and trifluoroacetic acid (3 mL) was stirred for 2 hours. The solvent was then removed in vacuo and the resulting residue dissolved in ethyl acetate. The ethyl acetate solution was washed with 1.0 N HCl (10 mL), dried over sodium sulfate, then filtered. Solvent was removed by rotary evaporator and the crude product purified by reverse phase preparative HPLC, (C18, 30%-40% CH3CN/H2O-0.1% CF3CO2H). Fractions containing product were concentrated by rotary evaporator and the resulting aqueous suspension extracted with ethyl acetate (3×50 mL). The combined organic layers were dried (sodium sulfate), filtered, and concentrated to dryness by rotary evaporator. The residue was triturated with ether and dried in vacuo to give the title compound as a white solid. 1H NMR (500 MHz, d6-Acetone) δ ppm: 11.96 (2H, s), 9.25 (2H, s), 8.98 (1H, br s), 7.23 (1H, t, J=7.6 Hz), 6.63 (1H, dd, J=8.2, 2.4 Hz), 6.57 (1H, dt, J=8.4, 2.4 Hz), 4.53 (2H, d, J=6.7 Hz), 4.05 (2H, d, J=5.2 Hz), 3.90 (2H, t, J=5.2 Hz), 1.55 (6H, s); HRMS [M+H]+ calcd for C17H19N3O5F: 364.13088; found: 364.1302.
Example 96




Carbamic acid, dimethyl-, 5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl ester. The title compound can be prepared from intermediate 158, dimethyl-carbamic acid 2-{[(3-benzyloxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydro-pyrimido[2,1-c][1,4]oxazine-2-carbonyl)-amino]-methyl}-5-fluoro-phenyl ester according to the method described for example 95. White crystalline powder; 1H NMR (CDCl3, 500 MHz) δ ppm: 1.56 (6H, s, Me), 2.96, 3.11 (2s, NMe), 3.99 (4H, s, CH2), 4.51 (2H, d, J=6 Hz, NCH2), 6.85 (1H, dd, J=2.5 Hz, 9 Hz, CH), 6.94 (1H, dt, J=2.5 Hz, 8.3 Hz, Ar—H), 7.37 (1H, dd, J=6.5 Hz, 8.5 Hz, Ar—H), 8.05 (1H, brt, J=5 Hz, NH), 12.0 (1H, s, OH); 13C NMR (CDCl3, 125.77 Hz) δ ppm: 27.90 (CH3), 36.68, 36.92 (2s, NCH3), 37.80 (NCH2), 43.14 (NCH2), 58.20 (OCH2), 75.99 (OC), 110.88, 110.07 (d, J=24 Hz, CH), 113.41, 113.57 (d, J=21 Hz, CH), 125.71 (C), 125.91, 125.94 (d, J=3.6 Hz, C), 131.57, 131.64 (d, J=9.6 Hz, CH), 146.22 (C), 150.83, 150.92 (d, J=11 Hz, C), 151.66 (C), 154.68 (C═O), 157.92 (C═O), 161.70, 163.68 (d, J=249 Hz, CF), 167.86 (C═O); HRMS calcd for C20H24N4O6F (M+H) 435.1680, found 435.1695 (δ+3.5 ppm). UV (MeOH): λmax 245 nm (ε1.05×104) 306 nm (ε8.00×103); Anal. calcd for C20H23N4O6F; C, 55.30; H, 5.34; N, 12.90: found C, 55.32; H, 5.38; N, 12.77.
Example 97




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[2-(methylamino)-2-oxoethoxy]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. A solution of intermediate 144, N-(4-fluoro-2-hydroxybenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.150 g, 0.331 mmol) and sodium hydride (0.015 g, 0.37 mmol, 60% oil dispersion) in anhydrous dimethylformamide (4 mL) was stirred for 5 minutes under a nitrogen atmosphere. The reaction mixture was treated with 2-chloro-N-methylacetamide (0.054 g, 0.50 mmol), and stirred for an additional 16 hours. Solvent was removed by rotary evaporator, and the resulting residue purified by short path flash silica gel chromatography (ethyl acetate). The fractions containing product were combined and concentrated to dryness. The residue was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL) and stirred for 1 hour. Solvent was removed by rotary evaporator and the crude product was triturated with a minimal volume of 95% ethanol. The resulting solid was collected by filtration, and dried under vacuum resulting in 93 mg (0.21 mmol, Yield 65%) of the title compound as a white powder: 1H NMR (500 MHz, CDCl3) δ ppm: 11.92 (1H, s), 7.67 (1H, t, J=6.3 Hz), 7.57 (1H, br), 7.30 (1H, dd, J=8.2, 6.4 Hz), 6.74 (1H, dt, J=8.2, 2.3 Hz), 6.60 (1H, dd, J=10.4, 2.4 Hz), 4.68 (2H, d, J=6.7 Hz), 4.45 (2H, s), 4.00 (4H, s), 2.93 (3H, d, J=0.9 Hz), 1.55 (6H, s). 13C NMR (125.77 MHz, CDCl3) δ ppm: 168.05, 167.44, 164.79, 162.82, 157.71, 156.57, 156.50, 152.14, 146.60, 131.94, 131.86, 125.23, 120.71, 120.68, 108.37, 108.20, 100.76, 100.55, 75.81, 67.46, 58.15, 43.24, 38.19, 28.08, 25.83. HRMS [M+H]+ calcd for C20H24N4O6F: 435.1680; found: 435.1668.
Example 98




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[2-(dimethylamino)-2-oxoethoxy]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 144, N-(4-fluoro-2-hydroxybenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide according to the method described for example 97. White powder; 1H NMR (500 MHz, CDCl3) δ ppm: 12.25 (1H, br s), 8.36 (1H, t, J=5.65 Hz), 7.31 (1H, dd, J=8.39, 6.56 Hz), 6.67 (1H, dt, J=8.32, 2.29 Hz), 6.57 (1H, dd, J=10.38, 2.14 Hz), 4.76 (2H, s), 4.62 (2H, d, J=6.10 Hz), 4.00 (4H, s), 3.06 (3H, s), 3.01 (3H, s), 1.58 (6H, s). 13C NMR (125.77 MHz, CDCl3) δ ppm: 168.25, 166.62, 164.23, 162.27, 158.02, 157.39, 151.36, 146.39, 131.25, 131.18, 125.95, 122.23, 122.20, 108.25, 108.08, 100.47, 100.27, 76.03, 66.24, 58.22, 43.15, 38.59, 36.07, 35.77, 27.94. HRMS [M+H]+ calcd for C21H26N4O6F: 449.18365; found: 449.1837.
Example 99




4-Morpholinecarboxylic acid, 5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl ester. The title compound can be prepared from intermediate 144, N-(4-fluoro-2-hydroxybenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide according to the method described for example 97. White powder; 1H NMR (500 MHz, CDCl3) δ ppm: 12.01 (1H, br s), 7.96 (1H, t, J=5.34 Hz), 7.38 (1H, dd, J=8.39, 6.26 Hz), 6.97 (1H, dt, J=8.24, 2.44 Hz), 6.87 (1H, dd, J=8.85, 2.44 Hz), 4.51 (2H, d, J=6.10 Hz), 4.00 (4H, s), 3.70-3.76 (4H, m), 3.65-3.70 (2H, m), 3.49-3.54 (2H, m), 1.55 (6H, s). 13C NMR (125.77 MHz, CDCl3) δ ppm: 167.91, 163.68, 157.86, 153.48, 151.71, 150.51, 150.42, 146.31, 131.58, 131.50, 125.83, 125.59, 113.84, 113.67, 111.03, 110.84, 75.93, 66.59, 66.52, 58.22, 45.17, 44.41, 43.16, 37.70, 27.95. HRMS [M+H]+ calcd for C22H26N4O7F: 477.17856; found: 477.1788.
Example 100




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(methylthio)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, and (4-fluoro-2-(methylthio)phenyl)methanamine according to the methods described for the synthesis of examples 1, 19 and 20. White solid. 1H NMR (300 MHz, CDCl3) δ ppm: 11.88 (1H, br), 8.03 (1H, t, J=6.04 Hz), 7.28 (1H, dd, J=8.42, 5.85 Hz), 6.93 (1H, dd, J=9.51, 2.20 Hz), 6.81 (1H, dt, J=8.23, 2.56 Hz), 4.58 (2H, d, J=6.22 Hz), 3.98 (4H, s), 2.49 (3H, s), 1.55 (6H, s); HRMS (ESI) calcd for C18H21FN3O4S (M+H) 394.1237, found 394.1218.
Example 101




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(methylsulfonyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. To a solution of example 100 pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(methylthio)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-(158 mg, 0.4 mmol), in CH2Cl2 (4 mL) was added 3-chloroperoxybenzoic acid (132 mg, 0.6 mmol; 77%, Aldrich) and the mixture stirred at room temperature for 2 hrs. After removing the solvent in vacuo, the residue was triturated with diethyl ether. The crude powder was purified by reverse phase column chromatography (YMC, ODS, 8% CH3CN/H2O-0.1% CF3CO2H) to provide 32 mg (0.075 mmol, Yield 19%) of the title compound as a white powder after trituration with diethyl ether, and 35 mg (0.086 mmol, Yield 21%) of the corresponding sulfoxide. 1H NMR (300 MHz, CDCl3) δ ppm: 11.71 (1H, s), 8.58 (1H, t, J=6.04 Hz), 7.73 (1H, dd, J=8.23, 2.74 Hz), 7.68 (1H, dd, J=8.42, 5.12 Hz), 7.32 (1H, dt, J=8.05, 2.93 Hz), 4.79 (2H, d, J=6.95 Hz), 3.97 (4H, s), 3.15 (3H, s), 1.56 (6H, s); HRMS (ESI) calcd for C18H19FN3O6S (M−H) 424.0979, found 424.0973.
Example 102




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(methylsulfinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo. Formed in the reaction described for example 101. 1H NMR (300 MHz, CDCl3) δ ppm: 11.73 (1H, s), 8.19 (1H, t, J=6.59 Hz), 7.54 (1H, dd, J=8.05, 2.93 Hz), 7.47 (1H, dd, J=8.42, 5.12 Hz), 7.16 (1H, dt, J=8.14, 2.74 Hz), 4.57-4.81 (2H, m), 3.99 (4H, s), 2.80 (3H, s), 1.56 (6H, s); HRMS (ESI) calcd for C18H21FN3O5S (M−H) 410.1196, found 410.1194.
Example 103




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(S-methylsulfinimidoyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. To a solution of example 100, pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(methylthio)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-, (245 mg, 0.75 mmol) in CH2Cl2 (3 mL) was added tert-butyl azido formate (115 mg, 0.8 mmol; prepared following the procedure described in Organic Synthesis 1979, 50, 9-12) and ferrous chloride (FeCl2, 50 mg) and the resulting mixture stirred for 18 hrs. The mixture was diluted with dichloromethane, washed with water, dried (MgSO4), filtered and concentrated to yield 450 mg of 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydro-pyrimido[2,1-c][1,4]oxazine-2-carboxylic acid 4-fluoro-2-(N-tert-butoxycarbonyl-5-methyl) sulfiliminyl-benzylamide as a dark gum; LC/MS m/z 509 (M+H).
A solution of this material, (100 mg) in CF3CO2H (1 mL), was stirred for 20 min then concentrated. The residue was purified by C-18 reverse phase HPLC (YMC ODS, 5-10% CH3CN/H2O-0.1% CF3CO2H) to provide 15 mg (0.037 mmol, Yield 21%) of the title compound as the corresponding trifluoroacetic acid salt. 1H NMR (300 MHz, DMSO-D6) δ ppm: 11.62 (1H, s), 9.57 (1H, t, J=6.0 Hz), 8.11 (1H, dd, J=8.8, 1.8 Hz), 7.62-7.71 (2H, m), 4.58-4.82 (2H, m), 3.97 (2H, t, J=4.8 Hz), 3.82 (2H, t, J=4.8 Hz), 3.37 (3H, s), 1.57 (6H, s); HRMS (ESI) calcd for C18H22FN4O4S (M+H) 409.1346, found 409.1333.
Example 104




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-[[3-[3-(trifluoromethyl)-3-diaziridinyl]-phenyl]-methyl]-. The title compound can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and (4-(diaziridin-3-yl)phenyl)methanamine (formed in the preparation of intermediate 66) according to the methods described for the synthesis of examples 1, 19 and 20. White solid; 1H NMR (300 MHz, CDCl3) δ ppm: 11.89 (1H, s), 7.81 (1H, t, J=6.0 Hz), 7.52-7.59 (2H, m), 7.38-7.44 (2H, m), 4.63 (2H, d, J=6.2 Hz), 4.00 (4H, s), 2.78 (1H, d, J=7.3 Hz), 2.21 (1H, d, J=8.1 Hz), 1.51-1.57 (6H, m); HRMS (ESI) calcd for C19H21F3N5O4 (M+H) 440.1546, found 440.1537.
Example 105




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-[[3-[3-(trifluoromethyl)-3H-diazirin-3-yl]-phenyl]methyl]-. The title compound can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and intermediate 66, (3-(3-(trifluoromethyl)diaziridin-3-yl)phenyl)methanamine, according to the methods described for the synthesis of examples 1, 19 and 20. White solid; 1H NMR (300 MHz, CDCl3) δ ppm: 11.86 (1H, s), 7.73-7.84 (1H, m), 7.35-7.40 (2H, m), 7.11-7.17 (1H, m, J=2.9 Hz), 7.08-7.11 (1H, m), 4.60 (2H, d, J=6.2 Hz), 4.00 (4H, s), 1.55 (6H, s); HRMS (ESI) calcd for C19H19F3N5O4 (M+H) 438.1389, found 438.1371.
Example 106




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(2-methyl-2H-tetrazol-5-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and intermediate 59, (4-fluoro-2-(2-methyl-2H-tetrazol-5-yl)phenyl)methanamine hydrochloride, according to the methods described for the synthesis of examples 1, 19 and 20. (white solid); 1H NMR (300 MHz, CDCl3) δ ppm: 12.01 (1H, s), 9.24 (1H, t, J=6.8 Hz), 7.75 (1H, dd, J=9.5, 2.6 Hz), 7.63 (1H, dd, J=8.4, 5.5 Hz), 7.15 (1H, dt, J=8.2, 2.6 Hz), 4.70 (2H, d, J=7.0 Hz), 4.45 (3H, s), 3.96 (4H, s), 1.53 (6H, s); HRMS (ESI) calcd for C19H21FN7O4 (M+H) 430.1639, found 430.1649.
Example 107




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1-methyl-1H-tetrazol-5-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and intermediate 62, (4-fluoro-2-(1-methyl-2H-tetrazol-5-yl)phenyl)methanamine hydrochloride, according to the methods described for the synthesis of examples 1, 19 and 20. Off-white solid; 1H NMR (500 MHz, CDCl3) δ ppm: 11.83 (1H, s), 9.17 (1H, t, J=5.8 Hz), 7.75 (1H, dd, J=8.4, 5.7 Hz), 7.30 (1H, t, J=8.2 Hz), 7.14 (1H, d, J=7.9 Hz), 4.43 (2H, d, J=6.7 Hz), 4.14-4.16 (3H, m), 4.00 (4H, s), 1.67 (6H, s); HRMS (ESI) calcd for C19H21FN7O4 (M+H) 430.1639, found 430.1619.
Examples 108-112
Examples 108-112 can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate and the indicated amines according to the methods described for the synthesis of examples 1, 19 and 20.
Example 108




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(dimethylamino)sulfonyl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 48, 2-(aminomethyl)-5-fluoro-N,N-dimethylbenzenesulfonamide. White solid; 1H NMR (500 MHz, CDCl3) δ ppm: 11.83 (1H, s), 8.64 (1H, t, J=6.6 Hz), 7.68 (1H, dd, J=8.6, 5.5 Hz), 7.50 (1H, dd, J=8.2, 2.8 Hz), 7.26-7.30 (1H, m), 4.80 (2H, d, J=7.0 Hz), 3.99 (4H, s), 2.91 (6H, s), 1.58 (6H, s); HRMS (ESI) calcd for C19H24FN4O6S (M+H) 455.1401, found 455.1402.
Example 109




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(methylamino)sulfonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 52, 2-(aminomethyl)-5-fluoro-N-methylbenzenesulfonamide hydrochloride. Off-white solid; 1H NMR (300 MHz, CDCl3) δ ppm: 11.69 (1H, s), 8.55 (1H, br), 7.58-7.67 (2H, m), 7.24-7.29 (1H, m), 4.87-4.97 (1H, br), 4.82 (2H, d, J=6.2 Hz), 3.97 (4H, s), 2.71 (3H, d, J=4.4 Hz), 1.56 (6H, s); HRMS (ESI) calcd for C18H22FN4O6S (M+H) 441.1244, found 441.1237.
Example 110




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(aminosulfonyl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 56, 2-(aminomethyl)-5-fluorobenzenesulfonamide hydrochloride. Off-white solid, 1H NMR (300 MHz, DMSO-D6) δ ppm: 11.92 (1H, s), 9.33 (1H, t, J=6.4 Hz), 7.66 (1H, dd, J=8.8, 2.2 Hz), 7.41-7.52 (2H, m), 4.89 (2H, d, J=6.2 Hz), 3.98 (2H, t, J=0.9 Hz), 3.83 (2H, t, J=4.9 Hz), 3.37 (2H, br), 1.55 (6H, s); HRMS (ESI) calcd for C17H20FN4O6S (M+H) 427.1088, found 427.1082.
Example 111




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(1-azetidinylsulfonyl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from (2-(azetidin-1-ylsulfonyl)-4-fluorophenyl)methanamine, which was synthesized according to the methods used to prepare intermediate 48. White solid, 1H NMR (500 MHz, CDCl3) δ ppm 11.86 (1H, s), 8.57 (1H, t, J=6.3 Hz), 7.65-7.72 (2H, m), 7.26-7.31 (1H, m), 4.82 (2H, d, J=6.7 Hz), 3.99 (4H, s), 3.96 (4H, t, J=7.8 Hz), 2.23-2.32 (2H, m), 1.58 (6H, s); HRMS (ESI) calcd for C20H24FN4O6S (M+H) 467.1401, found 467.1398.
Example 112




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(methylthio)phenyl]methyl]-4-oxo-. Prepared according to the method described for the synthesis of example 100. White solid, 1H NMR (300 MHz, CDCl3) δ ppm: 1.55 (6H, s), 2.50 (3H, s), 3.98 (4H, s), 4.65 (2H, d, J=6.6 Hz), 7.1-7.4 (3H, m), 8.11 (1H, t, J=5.9 Hz), 11.94 (1H, s); LC/MS m/z 376 (M+H).
Example 113




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(methylsulfinyl)phenyl]methyl]-4-oxo-. To a solution example 112, pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(methylthio)phenyl]methyl]-4-oxo-, (112 mg, 0.3 mmol) in CH2Cl2 (5 mL) was added 3-chloroperoxybenzoic acid (69 mg, 0.3 mmol; 77%, Aldrich) and the mixture stirred for 5 min. After removing the solvent in vacuo, the residue was purified by preparative reverse phase HPLC (YMC, ODS, 8-15% CH3CN/H2O-0.1% CF3CO2H) to provide 58 mg (0.16 mmol, Yield 53%) of the title compound as a white powder after trituration with diethyl ether. 1H NMR (300 MHz, CDCl3) δ ppm: 1.56 (6H, s), 2.80 (3H, s), 3.98 (4H, s), 4.60-4.94 (2H, m), 7.38-7.61 (3H, m), 7.65-7.89 (1H, m), 8.34 (1H, t, J=6.4 Hz), 11.82 (1H, s). HRMS (ESI) calcd for C18H22N3O5S (M+H) 392.1280, found 392.1281.
Example 114




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(methylsulfonyl)phenyl]methyl]-4-oxo-. To a solution of example 112, pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(methylthio)phenyl]methyl]-4-oxo-(112 mg, 0.3 mmol) in CH2Cl2 (5 mL) was added 3-chloroperoxybenzoic acid (140 mg, 0.62 mmol; 77%, Aldrich) and the mixture stirred for 20 hrs. After removing the solvent in vacuo, the residue was purified by preparative reverse phase HPLC(YMC, ODS, 15% CH3CN/H2O-0.1% CF3CO2H) to provide 61 mg (0.15 mmol, Yield 50%) of the title compound as a white powder after trituration with diethyl ether. 1H NMR (300 MHz, CDCl3) δ ppm: 1.56 (6H, s), 3.14 (3H, s), 3.96 (4H, s), 4.83 (2H, d, J=7.0 Hz), 7.39-7.57 (1H, m), 7.60-7.68 (2H, m), 8.02 (1H, d, J=8.4 Hz), 8.65 (1H, t, J=7.0 Hz), 11.78 (1H, brs). HRMS (ESI) calcd for C18H22N3O6S (M+H) 408.1229, found 408.1217.
Examples 115-116
Examples 115-116 can be prepared from the indicated intermediates by hydrogenolysis or trifluoroacetic acid mediated hydrolysis.
Example 115




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(2-thiazolylamino)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 163, N-(4-fluoro-2-(thiazol-2-ylamino)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (400 MHz, DMSO-d6) δ ppm: 11.97 (1H, brs), 9.72 (1H, brs), 9.39 (1H, t, J=6.3 Hz), 8.14 (1H, dd, J=2.5, 12.0 Hz), 7.31-7.28 (2H, m), 6.99 (1H, d, J=3.8 Hz), 6.84 (1H, ddd(dt), J=2.5, 8.3 Hz), 4.52 (2H, d, J=6.3 Hz), 3.96-3.94 (2H, m), 3.82-3.79 (2H, m), 1.54 (6H, s) LCMS (+ESI, M+H+) m/z 446. HRMS (ESI+) calculated for C20H21FN5O4S [M+H+]: 446.1298; found: 446.1292.
Example 116




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(5-methyl-1,3,4-thiadiazol-2-yl)amino]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 164, N-(4-fluoro-2-(5-methyl-1,3,4-thiadiazol-2-ylamino)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (400 MHz, DMSO-d6) δ ppm: 11.97 (1H, brs), 9.66 (1H, brs), 9.37 (1H, brt, J=6.1 Hz), 7.99 (1H, brd), 7.29 (1H, brt, J=7.1 Hz), 6.88 (1H, ddd(dt), J=2.5, 8.0 Hz), 4.52 (2H, d, J=6.1 Hz), 3.97-3.94 (2H, m), 3.82-3.80 (2H, m), 2.55 (3H, s), 1.54 (6H, s); LCMS (+ESI, M+H+) m/z 461. HRMS (ESI+) calculated for C20H22FN6O4S [M+H+]: 461.1407; found: 461.1425.
Example 117




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-A solution of intermediate 33, ethyl 2-(2-(3-chloropropoxy)propan-2-yl)-5-hydroxy-6-oxo-1,6-dihydropyrimidine-4-carboxylate, (0.208 g, 0.65 mmol) in anhydrous dimethylformamide (2 mL) was stirred with anhydrous potassium carbonate (0.366 g, 2.6 mmol) at 60° C. for 16 hours. This was treated with intermediate 69, (4-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride, (0.451 g, 2.08 mmol) and triethylamine (0.5 mL, 3.6 mmol) and stirring continued at 100° C. for 16 hours. The solvent was removed by rotary evaporator and the was purified by preparative reverse phase HPLC (C18, 10%-35% CH3CN/H2O-0.1% trifluoroacetic acid). Fractions containing product were pooled and concentrated by rotary evaporator. The resulting aqueous solution was extracted with ethyl acetate (2×50 mL), and the combined organic fractions dried (sodium sulfate), filtered, and concentrated to dryness. The resulting residue was triturated with a minimal volume of 95% ethanol, and the solid collected by filtration to give 103 mg (0.24 mmol, Yield 37%) of the title compound as a white solid: 1H NMR (500 MHz, CDCl3) δ ppm: 11.96 (1H, br s), 8.83 (1H, t, J=6.6 Hz), 8.46 (1H, s), 8.17 (1H, s), 7.71 (1H, dd, J=8.5, 6.1 Hz), 7.21 (1H, dt, J=8.2, 2.6 Hz), 7.11 (1H, dd, J=8.4, 2.6 Hz), 4.55 (2H, br), 4.44 (2H, d, J=6.7 Hz), 3.67 (2H, t, J=6.4 Hz), 1.91-1.97 (2H, p J=6.10 Hz), 1.63 (6H, s). 3C NMR (125.76 MHz, CDCl3) δ ppm: 167.97, 163.19, 161.20, 158.28, 153.35, 152.90, 147.34, 143.94, 136.89, 134.45, 134.37, 128.65, 128.62, 125.01, 117.09, 116.93, 112.42, 112.23, 82.46, 60.88, 39.13, 38.56, 27.73, 27.36. HRMS [M+H]+ calcd for C20H22N6O4F: 429.16867; found: 429.1687. Anal calcd for C20H21N6O4F.0.06H2O: C, 55.93; H, 4.96; N, 19.57; F, 4.42; found: C, 55.80; H, 5.14; N, 19.74; F, 4.46.
Example 118




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-[(methylamino)carbonyl]phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. The title compound can be prepared from intermediate 148, N-(4-fluoro-2-(methylcarbamoyl)benzyl)-3-(benzyloxy)-10,10-dimethyl-4-oxo-6,7,8,10-tetrahydro-4H-pyrimido[2,1-c][1,4]oxazepine-2-carboxamide according to the method described for example 46. White powder; 1H NMR (500 MHz, DMSO-D6) δ ppm: 12.17 (1H, s), 9.23 (1H, t, J=6.4 Hz), 8.56 (1H, q, J=0.3 Hz), 7.41 (1H, dd, J=8.6, 5.8 Hz), 7.34 (1H, dd, J=9.2, 2.8 Hz), 7.30 (1H, dt, J=8.6, 2.8 Hz), 4.55 (2H, d, J=6.4 Hz), 4.37 (2H, br), 3.64 (2H, t, J=6.4 Hz), 2.80 (3H, d, J=4.6 Hz), 1.80-1.86 (2H, m), 1.57 (6H, s); HRMS [M+H]+ calcd for C20H23N4O5F: 419.17308; found: 419.1713.
Example 119




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9,9-diethyl-N-[[4-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. The title compound can be prepared from intermediate 31, ethyl 9,9-diethyl-3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, and intermediate 69, (4-fluoro-2-(1H-1,2,4-triazol-1-yl)phenyl)methanamine hydrochloride, according to the methods described for examples 1, 19 and 20. White powder; 1H NMR (500 MHz, CDCl3) δ ppm: 11.89 (1H, br s), 8.81 (1H, t, J=6.6 Hz), 8.46 (1H, s), 8.15 (1H, s), 7.69 (1H, dd, J=8.6, 5.8 Hz), 7.22 (1H, dt, J=8.2, 2.6 Hz), 7.12 (1H, dd, J=8.4, 2.6 Hz), 4.45 (2H, d, J=6.7 Hz), 3.95-4.02 (4H, m), 1.95-2.03 (2H, m), 1.87-1.96 (2H, m), 0.86 (6H, t, J=7.5 Hz). 13C NMR (125.76 MHz, CDCl3) δ ppm: 167.98, 163.19, 161.20, 157.88, 152.81, 151.29, 146.05, 143.97, 137.00, 136.93, 134.28, 134.21, 128.62, 128.59, 125.85, 117.10, 116.94, 112.58, 112.38, 80.93, 58.63, 43.02, 39.20, 31.43, 7.86. HRMS [M+H]+ calcd for C21H24N6O4F: 443.18432; found: 443.1845.
Example 120




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9,9-diethyl-N-[[4-fluoro-2-[(methylamino)carbonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. A solution of intermediate 31, ethyl 9,9-diethyl-3-hydroxy-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, (0.228 g, 0.77 mmol) in anhydrous dimethylformamide (5 mL) was treated with anhydrous potassium carbonate (0.279 g, 2.0 mmol) followed by benzyl bromide (0.145 g, 0.85 mmol), and the mixture stirred for 16 hours. The mixture was treated with lithium hydroxide (0.042 g, 1.75 mmol) and water (2 mL) and stirred for 20 hours. The reaction mixture was diluted with water (30 mL) and brought to pH 1 with 6 N hydrochloric acid. The crude product was extracted with ethyl acetate (2×30 mL). The combined organic layers were dried (sodium sulfate), filtered, and concentrated to dryness.
A solution of this material, in anhydrous dimethylformamide (4 mL), was treated with HATU (0.32 g, 0.83 mmol) and stirred for 10 minutes. The reaction mixture was treated with intermediate 39, 2-aminomethyl-5-fluoro-N-methylbenzamide trifluoroacetic acid salt, (0.281 g, 0.94 mmol), followed by dimethylaminopyridine, DMAP, (0.140 g, 1.125 mmol), and stirred at 60° C. for 3 hours. Solvent was removed by rotary evaporator and the residue purified by flash column chromatography, eluting with 50% to 60% ethyl acetate in hexanes. Fractions containing the product were pooled and concentrated to dryness, which gave a white glassy solid. A sample (approximately 10 mg) was further dried under vacuum, and the remainder used in the following reaction: 1H NMR (500 MHz, CDCl3) δ ppm: 8.47 (1H, t, J=6.2 Hz), 7.36-7.45 (3H, m), 7.23-7.27 (2H, m), 7.10 (1H, dd, J=8.8, 2.9 Hz), 7.04 (1H, dt, J=8.3, 2.7 Hz), 6.60-6.69 (1H, br), 5.25 (2H, s), 4.50 (2H, d, J=6.2 Hz), 3.94 (4H, s), 2.95 (3H, d, J=5.1 Hz), 1.90-2.02 (5H, m), 0.75-0.82 (6H, m).
A solution of the above compound in dichloromethane (5 mL) and trifluoroacetic acid (5 mL) was stirred for 2 hours. The solvent was removed and the crude product was purified by preparative reverse phase HPLC (C-18 column, 10% to 40% CH3CN/H2O-0.1% CF3CO2H). Fractions containing product were pooled and concentrated in vacuo. The resulting aqueous suspension was extracted with dichloromethane (4×100 mL), and the combined organic extracts, dried (sodium sulfate), filtered, and concentrated to dryness. The residue was recrystallized ethanol/H2O to provide 0.012 g (0.028 mmol, Yield 4%) of the title compound as a white solid. 1H NMR (500 MHz, DMSO-D6) δ ppm: 12.10 (1H, s), 9.30 (1H, t, J=6.4 Hz), 8.52 (1H, m), 7.39 (1H, dd, J=8.4, 5.7 Hz), 7.26-7.35 (2H, m), 4.57 (2H, d, J=6.4 Hz), 3.94 (2H, t, J=0.9 Hz), 3.84 (2H, t, J=4.9 Hz), 2.79 (3H, d, J=0.6 Hz), 1.99-2.09 (2H, m), 1.81-1.91 (2H, m), 0.77 (6H, t, J=7.3 Hz). 13C NMR (125.76 MHz, DMSO-D6) δ ppm: 167.86, 167.57, 159.65, 156.86, 151.21, 145.10, 137.26, 137.21, 132.52, 130.75, 130.68, 125.39, 116.69, 116.52, 114.71, 114.53, 80.04, 57.60, 42.62, 40.05, 30.04, 26.03, 7.47. HRMS [M+H]+ calcd for C21H26N4O5F: 433.18873; found: 433.1872.
Examples 121-130
Examples 121-130 can be prepared from the indicated intermediates according to the methods described for examples 121 and 122. Alternatively, the examples can be formed by treating the indicated intermediates with trifluoroacetic acid.
Example 121




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(2-oxo-1-piperidinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. To intermediate 145, N-(4-fluoro-2-(2-oxopiperidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazin-4(9H)-one, (100 mg, 0.187 mmol) in ethyl acetate (10 mL) was added palladium (10% on charcoal) (30 mg). The reaction mixture was stirred at 23° C. under a hydrogen atmosphere (balloon) for 3 h. The catalyst was removed by filtration on Celite. The filtrate was concentrated in vacuo and the resulting residue triturated with diethyl ether (10 ml) and dried in vacuo to afford 37 mg (45% yield) of the title compound. IR (KBr, cm−1) 3397, 2943, 1636, 1539, 1173. 1HNMR 400 MHz (MeOD)6 ppm: 7.51 (1H, dd,(t), J=7.0 Hz), 7.11 (2H, m), 4.70 (1H, d, J=15.3 Hz), 4.24 (1H, d, J=15.3 Hz), 4.05 (2H, m), 3.96 (2H, m,), 3.73 (1H, m), 3.62 (1H, m), 2.63-2.48 (2H, m), 2.03 (4H, broad s), 1.62 (6H, s). LCMS (M+H)+ m/z 445.
Example 122




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-[[2-(2-oxo-3-oxazolidinyl)phenyl]methyl]-. To intermediate 161, N-(4-fluoro-2-(2-oxopiperidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazin-4(9H)-one (100 mg, 0.187 mmol) in ethyl acetate (10 mL) was added palladium (10% on charcoal) (30 mg). The reaction mixture was stirred at 23° C. under a hydrogen atmosphere (balloon) for 3 h. The catalyst was removed by filtration on Celite. The filtrate was concentrated in vacuo and the resulting residue triturated with diethyl ether (10 ml) and dried in vacuo to afford 37 mg (45%) of the title compound. IR (KBr, cm−1) 3397, 2943, 1636, 1539, 1173. 1HNMR 400 MHz (MeOD) δ ppm: 7.51 (1H, dd, (t), J=7.0 Hz), 7.11 (2H, m), 4.70 (1H, d, J=15.3 Hz), 4.24 (1H, d, J=15.3 Hz), 4.05 (2H, m), 3.96 (2H, m,), 3.73 (1H, m), 3.62 (1H, m), 2.63-2.48 (2H, m), 2.03 (4H, broad s), 1.62 (6H, s). LCMS (M+H)+ m/z 445.
Example 123




Pyrimido[2,1-c][1, 4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-[[2-(2-oxo-1-azetidinyl)phenyl]methyl]-. The title compound can be prepared from intermediate 162, N-(2-(2-oxoazetidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (400 MHz, CDCl3) δ ppm: 12.15 (1H, brs), 9.65 (1H, brs), 7.33-7.20 (4H, m), 4.54 (2H, d, J=6.6 Hz), 3.96-3.94 (2H, m), 3.82-3.79 (2H, m), 3.76 (2H, t, J=4.3 Hz), 3.10 (2H, t, J=4.3 Hz), 1.53 (6H, s); LCMS (+ESI, M+H+) m/z 399.
Example 124




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(2-oxo-1-pyrrolidinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo. The title compound can be prepared from intermediate 159, N-(4-fluoro-2-(2-oxopyrrolidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. IR (KBr, cm−1) 3432, 2980, 1689, 1543, 1183. 1HNMR 400 MHz (MeOD) δ ppm: 7.53 (1H, dd, J=9.2, 6.3 Hz), 7.11 (2H, m), 4.52 (2H, s), 4.05 (2H, t, J=5.0 Hz), 3.96 (2H, m), 3.87 (2H, t, J=7.1 Hz), 2.61 (2H, t, J=8.0 Hz), 2.26 (2H, m), 1.6 (6H, s).HRMS (ESI+) calculated for C21H24FN4O5 [M+H+]: 431.1731; found: 431.1714.
Example 125




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(hexahydro-2-oxo-1H-azepin-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 160, N-(4-fluoro-2-(2-oxoazepan-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2 carboxamide. IR (KBr, cm−1) 3392, 2934, 1646, 1522, 1292. 1HNMR 400 MHz (MeOD) δ ppm: 7.50 (1H, broad s), 7.06 (1H, broad s), 7.01 (1H, d, J=9.0 Hz), 4.64 (1H, d, JAB=15.0 Hz), 4.31 (1H, d, JAB=15 Hz), 4.04 (2H, m), 3.96 (3H, m), 3.66 (1H, m), 2.86 (1H, m), 2.65 (1H, m), 2.03-1.80 (6H, m), 1.62 (6H, s), (2H, m). LCMS (M+H)+ m/z 459.
Example 126




N-(4-Fluoro-2-(2-oxoazetidin-1-yl)benzyl)-3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 146, N-(4-fluoro-2-(2-oxoazetidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (400 MHz, DMSO-d6) δ: 9.46 (d, J=6.4 Hz), 7.34 (1H, dd, J=8.5, 6.4 Hz), 7.25 (1H, dd, J=10.2, 2.6 Hz), 7.08 (1H, td, J=8.5, 2.6 Hz), 6.90 (1H, m), 4.52 (2H, d, J=6.4 Hz), 3.96 (2H, t, J=5.0 Hz), 3.83-3.77 (4H, m), 3.11 (2H, t, J=5.0 Hz), 1.53 (6H, s); LCMS (M+H)+ m/z 417.
Example 127




N-(4-Fluoro-2-(2-oxooxazolidin-3-yl)benzyl)-3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared from intermediate 165, N-(4-fluoro-2-(2-oxooxazolidin-3-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (400 MHz, DMSO-d6) δ ppm: 12.12 (1H, s), 9.34 (1H, t, J=6.2 Hz), 7.41 (1H, m), 7.23 (1H, td, J=8.6, 2.5 Hz), 4.48 (4H, m), 4.06 (2H, t, J=7.7 Hz), 3.97 (2H, t, J=5.0 Hz), 3.83 (2H, t, J=5.0 Hz), 1.55 (6H, s). LCMS (M+H)+ m/z 433.
Example 128




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(2R)-2-(hydroxymethyl)-5-oxo-1-pyrrolidinyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 192, (R) —N-(4-fluoro-2-(2-(hydroxymethyl)-5-oxopyrrolidin-1-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. IR (KBr, cm−1) 3441, 2979, 1684, 1540, 1172. 1HNMR 400 MHz (MeOD) δ ppm: 7.56 (1H, m), 7.15 (2H, m), 4.86-4.31 (3H, m), 4.06 (2H, t, 5.0 Hz), 3.98 (2H, t, 5.0 Hz), 3.57 (2H, broad s), 2.72-2.55 (2H, m), 2.41 (1H, m), 2.25 (1H, m), 1.62 (6H, s). LCMS (M+H)+ m/z 461.
Example 129




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(2R)-2-[(acetyloxy)methyl]-5-oxo-1-pyrrolidinyl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 193, (R)-(1-(2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenyl)-5-oxopyrrolidin-2-yl)methyl acetate. 1HNMR 400 MHz (MeOD) δ ppm: 7.58 (1H, m), 7.21-7.04 (2H, m), 5.24 (2H, s), 4.66-4.21 (3H, m), 4.02-3.92 (5H, m), 2.73-2.34 (3H, m), 2.23-1.61 (5H, m), 1.60 (6H, s). LCMS (M+H)+ m/z 503.
Example 130




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(2S)-2-(hydroxymethyl)-5-oxo-1-pyrrolidinyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 167, (S)—N-(2-(2-((tert-butyldimethylsilyloxy)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (DMSO) δ ppm: 7.36 (1H, m), 7.18 (2H, m), 5.07 (1H, broad s), 4.66-4.11 (3H, m), 3.95 (2H, t, 5.1 Hz), 3.80 (2H, t, 5.1 Hz), 3.39 (2H, s), 2.43 (1H, m), 2.25 (1H, m), 2.08 (1H, m), 1.51 (6H, s). LCMS (M+H)+ m/z 461.
Example 131




(R)—N-(2-(2-((Dimethylamino)methyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of intermediate 195, (R)—N-(2-(2-(azidomethyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, (0.050 g, 0.087 mmol) in MeOH (5 mL) was added Palladium (10%) on charcoal (0.020 g) and formaldehyde (0.30 mL, 10 mmol). The reaction mixture was stirred at 23° C. under H2 (balloon) for 5 h. Palladium on charcoal was then removed by filtration and the solvent was evaporated in vacuo. The residue was purified by preparative HPLC (YMC-Pack C-18) to afford the title compound (0.013 g, 29% yield): 1H NMR (400 MHz, MeOD) δ ppm: 7.56 (1H, dd, J=7.0 Hz), 7.30-7.17 (2H, m), 4.70 (2H, m), 4.15 (1H, d, J=15.3 Hz), 4.07 (2H, m), 3.99 (2H, m,), 3.54 (1H, m), 2.93 (6H, s), 2.84-2.64 (4H, m), 2.20 (1H, m), 1.65 (6H, s). HRMS calcd for C24H31N5O5F: 488.2309; found 488.2328.
Example 132




(R)—N-(2-(2-(Azidomethyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of intermediate 195, (R)—N-(2-(2-(azidomethyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, (0.030 g, 0.052 mmol) in CH2Cl2 (1 mL) was added CF3CO2H (1 mL) at 23° C. The reaction mixture was stirred at 23° C. for 3 h. Toluene (20 mL) was then added and the solvents evaporated in vacuo. The residue was purified by preparative HPLC (YMC-Pack C-18) to afford the title compound (0.008 g, 31%): 1H NMR (400 MHz, MeOD) δ ppm: 7.55 (1H, dd, J=6.6 Hz), 7.16 (2H, m), 4.47-4.24 (2H, m), 4.05 (2H, m), 3.98 (2H, m), 3.56 (2H, s), 2.70-2.45 (4H, m), 2.11 (1H, m), 1.64 (3H, s), 1.61 (3H, s). HRMS calcd for C22H25N7O5F: 486.1901; found 486.1923.
Example 133




(R)—N-(2-(2-(Aminomethyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. The title compound can be prepared by hydrogenolysis (H2, 10% Pd—C) of intermediate 195, (R)—N-(2-(2-(azidomethyl)-5-oxopyrrolidin-1-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1H NMR (400 MHz, MeOD) δ ppm: 7.54 (1H, dd, J=7.0 Hz), 7.25-7.13 (2H, m), 4.62 (2H, m), 4.17 (1H, d, J=15.6 Hz), 4.07 (2H, m), 3.99 (2H, m,), 3.23 (2H, d, J=3.1 Hz), 2.79-2.59 (3H, m), 2.14 (1H, m), 1.66 (6H, s). HRMS calcd for C22H27N5O5F: 460.1996; found 460.2014.
Examples 134-136
Examples 134-136 can be prepared from the indicated intermediates by hydrogenolysis or trifluoroacetic acid mediated hydrolysis.
Example 134




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-amino-4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 169, N-(2-amino-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. IR (KBr, cm−1) 3383, 2979, 1636, 1540, 1288. 1HNMR 400 MHz (MeOD) δ ppm: 7.14 (1H, dd, J=7.3 Hz), 6.44-6-31 (2H, m), 4.69 (1H, broad s), 4.48 (2H, broad s), 4.03 (2H, t, J=4.6 Hz), 3.95 (2H, t, J=4.6 Hz), 1.62 (6H, s). HRMS (ESI+) calculated for C17H20FN4O4 [M+H+]:363.1469; found: 363.1454.
Example 135




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(acetylamino)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 189, N-(2-acetamido-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (MeOD) δ ppm: 7.66 (1H, broad d, J=10.5 Hz), 7.40 (1H, broad t, 6.5 Hz), 6.87 (1H, broad t, J=6.5 Hz), 4.55 (2H, m), 4.02 (2H, t, J=5.0 Hz), 3.92 (2H, t, J=5.0 Hz), 2.28 (3H, s), 1.62 (6H, s). HRMS (ESI+) calculated for C19H22FN4O5 [M+H+]:405.1574; found: 405.1571.
Example 136




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(acetylmethylamino)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 168, N-(4-fluoro-2-(N-methylacetamido)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. IR (KBr, cm−1) 3407, 2979, 1653, 1539, 1116. 1HNMR 400 MHz (MeOD) δ ppm: 7.53 (1H, m), 7.24-7.06 (2H, m), 4.61-4.45 (2H, m), 4.08 (2H, t, J=5.1 Hz), 4.00 (2H, t, J=5.1 Hz), 3.40 (0.6H, s), 3.25 (2.4H, s), 1.85 (2.4H, s), 1.81 (0.6H, s), 1.65 (2.4H, s), 1.64 (2.4H, s), 1.62 (1.2H, s). LCMS (M+H)+ m/z 419.
Example 137




N-(2-(2,5-Dioxo-2H-pyrrol-1(5H)-yl)benzyl)-3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. To a solution of N-(2-aminobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido [2,1-c][1,4]oxazine-2-carboxamide (0.050 g, 0.114 mmol) in acetic acid (1.5 mL) was added maleic anhydride (0.013 g, 0.131 mmol) and the reaction mixture stirred in a sealed tube at 115 C for 18 h. Acetic acid was evaporated in vacuo and the residue was purified by preparative HPLC (YMC-Pack C-18) to afford the title compound (0.013 g, 27% yield): 1H NMR (400 MHz, CDCl3) δ ppm: 8.03 (1H, broad t), 7.59 (1H, m), 7.48 (2H, m), 7.19 (1H, m), 6.92 (2H, s), 4.48 (2H, d, J=6.1 Hz), 4.04 (4H, s), 1.62 (6H, s); HRMS calcd for C21H21N4O6: 425.1461; found 425.1451.
Example 138




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-(benzo[b]thien-7-ylmethyl)-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. To intermediate 147, N-(benzo[b]thiophen-7-ylmethyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, (60 mg, 0.13 mmol) was added CF3CO2H (2 mL) at 23° C. The reaction mixture was stirred at 23° C. for 1.5 h. Toluene (20 mL) was then added and the solvents were evaporated in vacuo. The residue was purified by preparative HPLC (YMC-Pack C-18) to afford the title compound (0.017 g, 34% yield): 1H NMR (400 MHz, CDCl3) δ: 8.00 (1H, broad t), 7.86 (1H, d, J=8.0 Hz), 7.51-7.28 (3H, m), 4.93 (2H, d, J=3.9 Hz), 4.05 (4H, s), 1.58 (6H, s). HRMS calcd for C19H20N3O4S: 386.1175; found 386.1165.
Example 139




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(1,1-dioxidobenzo[b]thien-7-yl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. To intermediate 196, N-(benzo[b]thiophen-1,1-dione-7-ylmethyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-. tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide, (0.080 g, 0.16 mmol) was added CF3CO2H (2 mL) at 23° C. The reaction mixture was stirred at 23° C. for 2.5 h. Toluene (20 mL) was then added and the solvents evaporated in vacuo. The residue was purified by crystallization with MeOH (10 mL) to afford the title compound (0.037 g, 55% yield). 1H NMR (400 MHz, DMSO-d6) δ: 11.87 (1H, s), 9.44 (1H, t, J=6.5 Hz), 7.65 (2H, m), 7.51 (1H, d, J=7.0 Hz), 7.42-7.37 (2H, m), 4.84 (2H, d, J=6.2 Hz), 3.99 (2H, t, 5.4 Hz), 3.85 (2H, t, J=5.4 Hz), 1.59 (6H, s). HRMS calcd for C19H20N3O6S: 418.1073; found 418.1078.




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2,3-dihydro-1,1-dioxidobenzo[b]thien-7-yl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound was obtained from intermediate 196, N-(benzo[b]thiophen-1,1-dione-7-ylmethyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide by reduction, Palladium (10%) on charcoal under H2. 1HNMR (400 MHz, CDCl3) δ: 8.65 (1H, s), 7.58-7.49 (2H, m), 7.32 (1H, d, J=7.0 Hz), 4.86 (2H, d, J=6.8 Hz), 3.99 (4H, s), 3.54 (2H, t, J=6.8 Hz), 3.40 (2H, t, J=6.8), 1.61 (6H, s). LCMS (M+H)+ m/z 420.
Examples 141-146
Examples 141-146, listed in Table 9, can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, according to the methods described for examples 1, 19, and 20. The compounds in the table were characterized by LCMS; (Xterra MS-C18, 4.6×50 mm); eluted with 10% to 100% B, 4.5 min gradient, (A=H2O-0.1% CF3CO2H, B=CH3CN-0.1% CF3 CO2H); flow rate at 2.5 mL/min. UV detection at 220 nm). Product retention time (RT, minutes) and molecular weight (MS [M+1]) results are listed in the table.










TABLE 9





Ex-





am-


ple
Structure
RT
MS


















141




1.1
331.17



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-(2-pyridinylmethyl)-


 


142




2.07
346.18



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[(5-methylpyrazinyl)methyl]-4-oxo-


 


143




3.35
376.16



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[4-(methylthio)phenyl]methyl]-4-oxo-


 


144




2.37
397



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[(2,3-dimethyl-1H-indol-5-yl)methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-


 


145




2.54
388.2



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[4-(1-methylethoxy)phenyl]methyl]-4-oxo-


 


146




3.23
366.15



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[(3,5-difluorophenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-









Example 147




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-bromophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 25, ethyl 3-hydroxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxylate, and 2-bromobenzylamine according to the methods described for examples 1, 19 and 20. 1H NMR (300 MHz, DMSO-D6) δ ppm: 1.58 (s, 1), 3.84 (t, 2), 3.98 (t, 2), 4.54 (d, 2), 7.25 (m, 2), 7.38 (m, 2), 7.64 (d, 1), 9.43 (brm, 1), 12.01 (s, 1).
Examples 148-200
Examples 148-200, listed in table 10, were synthesized according to the following procedure. To a microwave reaction vessel containing a stir bar, was added Pd(Ph3P)4 (30 mg, 25 μmol), followed by anhydrous dioxane (0.5 mL). To this was added example 147, pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-bromophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-, (50 μmol), boronic acid or boronate ester reagent (200 μmol), anhydrous dioxane (0.5 mL) and 2M K3PO4 aqueous solution (0.25 mL). The reaction vessel was flushed with nitrogen, capped and heated at 120° C. for 10 minutes in a microwave reactor. The reaction mixture was filtered through a Whatman 0.45 μm syringe filter and the crude product purified using preparative HPLC (Xterra MS-C18, 30×50 mm); Eluted with 30% to 100% B, 8 min gradient, (A=10 mM NH4OAC (aq.), B=CH3CN); flow rate at 30 mL/min. UV detection at 220 nm) to give the title compound.
The compounds in the table were characterized by LCMS; (Xterra MS-C18, 4.6×50 mm); eluted with 10% to 100% B, 4.5 min gradient, (A=H2O-0.1% CF3CO2H, B=CH3CN-0.1% CF3CO2H); flow rate at 2.5 mL/min. UV detection at 220 nm). Product retention time (RT, minutes) and observed molecular weight (MS [M+1]) results are listed in the table.










TABLE 10





Example
Structure
RT
MS


















148




2.08
457



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(8-quinolinyl)phenyl]methyl]-


 


149




1.91
467.95



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(2,4-dimethoxy-5-



pyrimidinyl)phenyl]methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


150




1.71
407



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(3-pyridinyl)phenyl]methyl]-


 


151




1.98
425.01



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(3-pyridinyl)phenyl]methyl]-


152




2.04
437.04



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[[2-(2-



methoxy-3-pyridinyl)phenyl]methyl]-9,9-



dimethyl-4-oxo-


 


153




2.1
436.97



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[[2-(5-



methoxy-3-pyridinyl)phenyl]methyl]-9,9-



dimethyl-4-oxo-


154




2.41
467.01



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(2,6-dimethoxy-3-



pyridinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


155




2.19
486.03



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-[1-(phenylmethyl)-1H-pyrazol-4-



yl]phenyl]methyl]-


 


156




2
492.01



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[2-[6-(4-morpholinyl)-3-



pyridinyl]phenyl]methyl]-4-oxo-


 


157




3.02
454.97



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(6-amino-3-pyridinyl)phenyl]methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-


 


158




1.99
438.04



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(1-propyl-1H-pyrazol-4-



yl)phenyl]methyl]-


 


159




1.66
505.05



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[2-[2-(4-methyl-1-piperazinyl)-4-



pyridinyl]phenyl]methyl]-4-oxo-


 


160




3.2
406



Pyrimido [2,1-c][1,4]oxazine-2-carboxamide,



N-([1,1′-biphenyl]-2-ylmethyl)-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


161




3.36
469.91



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[(4′-chloro-2′-methoxy[1,1′-biphenyl]-2-



yl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-



dimethyl-4-oxo-


 


162




3.39
419.97



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[(4′-methyl[1,1′-biphenyl]-2-yl)methyl]-4-oxo-


 


163




3.36
419.97



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[(3′-methyl[1,1′-biphenyl]-2-yl)methyl]-4-oxo-


 


164




3.14
435.97



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[(3′-



methoxy[1,1′-biphenyl]-2-yl)methyl]-9,9-



dimethyl-4-oxo-


 


165




2.65
421.99



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[(3′-



hydroxy[1,1′-biphenyl]-2-yl)methyl]-9,9-



dimethyl-4-oxo-


 


166




2.91
430.99



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[(4′-cyano[1,1′-biphenyl]-2-yl)methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-


 


167




2.93
430.99



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[(3′-cyano[1,1′-biphenyl]-2-yl)methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-


 


168




3.02
491



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[4′-(4-morpholinyl)[1,1′-biphenyl]-2-



yl]methyl]-4-oxo-


 


169




2.11
435.02



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[4′-(4-morpholinyl)[1,1′-biphenyl]-2-



yl]methyl]-4-oxo-


 


170




2.54
476.96



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[4′-[(dimethylamino)carbonyl][1,1′-



biphenyl]-2-yl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


171




2.53
476.96



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[3′-[(dimethylamino)carbonyl][1,1′-



biphenyl]-2-yl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


172




2.66
502.97



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[3′-(1-pyrrolidinylcarbonyl)[1,1′-



biphenyl]-2-yl]methyl]-


 


173




2.26
448.94



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[4′-(aminocarbonyl)[1,1′-biphenyl]-2-



yl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-



dimethyl-4-oxo-


 


174




2.27
448.94



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[3′-(aminocarbonyl)[1,1′-biphenyl]-2-



yl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-



dimethyl-4-oxo-


 


175




2.36
462.98



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[4′-[(methylamino)carbonyl][1,1′-biphenyl]-



2-yl]methyl]-4-oxo-


 


176




2.63
498.94



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[3′-[(methylsulfonyl)amino][1,1′-biphenyl]-2-



yl]methyl]-4-oxo-


 


177




3
444.97



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[[2-(1H-indol-



5-yl)phenyl]methyl]-9,9-dimethyl-4-oxo-


 


178




2.56
462.98



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[4′-[(dimethylamino)methyl][1,1′-



biphenyl]-2-yl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


179




3.09
411.98



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(3-thienyl)phenyl]methyl]-


 


180




2.66
425.01



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(3,5-dimethyl-4-



isoxazolyl)phenyl]methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


181




3.08
450.01



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[[4′-



(methoxymethyl)[1,1′-biphenyl]-2-yl]methyl]-



9,9-dimethyl-4-oxo-


 


182




2.43
438



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[[2-(2-



methoxy-5-pyrimidinyl)phenyl]methyl]-9,9-



dimethyl-4-oxo-


 


183




1.71
410



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[2-(1-methyl-1H-pyrazol-4-



yl)phenyl]methyl]-4-oxo-


 


184




3.02
455



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(5-fluoro-6-methoxy-3-



pyridinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


185




3.14
436



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-N-[(4′-



methoxy[1,1′-biphenyl]-2-yl)methyl]-9,9-



dimethyl-4-oxo-


 


186




3.30
478



[1,1′-Biphenyl]-4-carboxylic acid, 2′-



[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-



4-oxopyrimido[2,1-c][1,4]oxazin-2-



yl)carbonyl]amino]methyl]-,ethyl ester


 


187




2.76
450.2



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(1,3-benzodioxol-5-yl)phenyl]methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-


 


188




2.92
442.13



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[(3′,4′-difluoro[1,1′-biphenyl]-2-yl)methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-


 


189




2.57
445.15



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[4′-(cyanomethyl)[1,1′-biphenyl]-2-



yl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-



dimethyl-4-oxo-


 


190




1.34
407.34



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(4-pyridinyl)phenyl]methyl]-


 


191




1.40
421.2



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[2-(4-methyl-3-pyridinyl)phenyl]methyl]-4-



oxo-


 


192




2.57
439.36



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(6-fluoro-5-methyl-3-



pyridinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


193




2.84
471.13



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(5-chloro-6-methoxy-3-



pyridinyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


194




1.62
457.26



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(3-quinolinyl)phenyl]methyl]-


 


195




1.57
520.31



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[3′-[[[2-



(dimethylamino)ethyl]amino]carbonyl][1,1′-



bzphenyl]-2-yl]methyl]-4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-


 


196




1.57
532.41



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-



[[4′-[(4-methyl-1-piperazinyl)carbonyl][1,1′-



biphenyl]-2-yl]methyl]-4-oxo-


 


197




1.55
457.26



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-N-[[2-(5-quinolinyl)phenyl]methyl]-


 


198




2.86
396.2



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(3-furanyl)phenyl]methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


199




3.08
451.2



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(6-ethoxy-3-pyridinyl)phenyl]methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-


 


200




3.33
396.34



Pyrimido[2,1-c][1,4]oxazine-2-carboxamide,



N-[[2-(1-cyclopenten-1-yl)phenyl]methyl]-



4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-



oxo-









Example 201




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluoro-2,5-dibromophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo. A mixture of (2,5-dibromo-4-fluorophenyl)methanamine (1 mmol), intermediate 25 (0.268 g, 1 mmol) and triethylamine (0.5 mL, 3.5 mmol) in ethanol/dimethylformamide (1:1, 3 mL) was heated at 100° C. for 6 h. After cooling, the reaction mixture was purified by preparative HPLC to afford the title compound (0.31 g, 62% yield) as an off-white solid. 1H NMR (500 MHz, CDCl3) δ ppm: 11.72 (1H, s), 8.08 (1H, t, J=6.1 Hz), 7.61 (1H, d, J=7.0 Hz), 7.38 (1H, d, J=7.6 Hz), 4.61 (2H, d, J=6.4 Hz), 4.02 (4H, s), 1.59 (6H, s).
Example 202




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-(methylsulfonyl)phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. Off-white solid. 1H NMR (500 MHz, CDCl3) δ ppm: 11.77 (1H, br s), 8.55 (1H, t, J=6.3 Hz), 7.75 (1H, dd, J=8.1, 2.6 Hz), 7.72 (1H, dd, J=8.5, 5.2 Hz), 7.34 (1H, td, J=8.0, 2.6 Hz), 4.81 (2H, d, J=6.7 Hz), 4.54 (2H, br), 3.65 (2H, t, J=6.1 Hz), 3.18 (3H, s), 1.93 (2H, m), 1.59 (6H, s); 13C NMR (126 MHz, CDCl3) δ ppm: 168.14, 163.14, 161.13, 158.23, 153.73, 147.26, 140.77, 140.72, 135.31, 135.25, 132.83, 132.80, 124.83, 121.69 121.52, 117.63, 117.43, 82.56, 60.80, 45.14, 40.24, 38.62, 27.76, 27.35. HRMS (ESI) calcd for C19H23N3O6FS (M+H): 440.1292; found: 440.1300. Anal. Calcd for C19H22N3O6FS.0.06H2O: C, 51.80; H, 5.06; N, 9.54; F, 4.31; found C, 51.83; H, 4.97; N, 9.29; F, 4.12.
Example 203




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9,9-diethyl-N-[[4-fluoro-2-(methylsulfonyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. 1H NMR (500 MHz, CDCl3) δ ppm: 11.74 (1H, s), 8.54 (1H, t, J=6.6 Hz), 7.75 (1H, dd, J=8.2, 2.7 Hz), 7.71 (1H, dd, J=8.5, 5.2 Hz), 7.34 (1H, td, J=8.0, 2.6 Hz), 4.82 (2H, d, J=7.0 Hz), 3.94-4.00 (4H, m), 3.17 (3H, s), 1.93-2.00 (2H, m), 1.84-1.92 (2H, m), 0.83 (6H, t, J=7.3 Hz); 13C NMR (126 MHz, CDCl3) δ ppm: 168.26, 163.13, 161.11, 157.83, 151.66, 146.05, 140.69, 140.64, 135.13, 135.07, 132.80, 132.77, 125.63, 121.65, 121.48, 117.65, 117.45, 81.03, 58.51, 45.14, 43.08, 40.33, 31.38, 7.87. HRMS (ESI) calcd for C20H25N3O6FS (M+H): 454.1448; found: 454.1448. Anal. calcd for C20H24N3O6FS: C, 52.97; H, 5.33; N, 9.27; S, 7.07; found: C, 53.01; H, 5.60; N, 9.10; S, 7.00.
Example 204




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[2-[(dimethylamino)sulfonyl]-4-fluorophenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. Pale orange solid. 1H NMR (500 MHz, CDCl3) δ ppm: 11.87 (1H, br s), 8.59 (1H, t, J=6.4 Hz), 7.69 (1H, dd, J=8.5, 5.2 Hz), 7.49 (1H, dd, J=8.2, 2.4 Hz), 7.26 (1H, td, J=7.7, 3.2 Hz), 4.79 (2H, d, J=6.7 Hz), 4.53 (2H, br), 3.65 (2H, t, J=6.1 Hz), 2.90 (6H, s), 1.89-1.96 (2H, m), 1.57-1.61 (6H, s); 13C NMR (126 MHz, CDCl3) δ ppm: 168.03, 162.71, 160.70, 158.29, 153.57, 147.23, 138.51, 138.46, 135.30, 135.25, 132.70, 132.67, 124.99, 120.40, 120.24, 116.93, 116.73, 82.59, 60.79, 58.57, 40.26, 38.58, 37.59, 27.74, 27.36, 18.55. HRMS (ESI) calcd for C20H26N4O6FS (M+H) 469.1557; found 469.1557. Anal. calcd for C20H26N4O6FS.CH3CH2OH: C, 51.29; H, 6.07; N, 10.89; S, 6.01; F, 3.69; found C, 51.29; H, 6.33; N, 10.85; S, 6.01; F, 3.54.
Example 205




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(dimethylamino)sulfonyl]-4-fluorophenyl]methyl]-9,9-diethyl-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. White glassy solid. 1H NMR (500 MHz, CDCl3) δ ppm: 11.85 (1H, br), 8.59 (1H, t, J=6.4 Hz), 7.68 (1H, dd, J=8.5, 5.2 Hz), 7.49 (1H, dd, J=8.2, 2.4 Hz), 7.24-7.30 (1H, m), 4.80 (2H, d, J=7.0 Hz), 3.94-4.01 (4H, m), 2.90 (6H, s), 1.93-2.01 (2H, m), 1.85-1.93 (2H, m), 0.83 (6H, t, J=7.3 Hz); 13C NMR (126 MHz, CDCl3) δ ppm: 168.04, 162.70, 160.70, 158.06, 151.44, 145.95, 138.53, 138.49, 135.17, 135.12, 132.62, 132.59, 125.95, 120.37, 120.21, 116.87, 116.68, 81.08, 58.51, 43.11, 40.32, 37.58, 31.35, 7.87. HRMS (ESI) calcd for C21H28N4O6FS (M+H) 483.1714; found 483.1702. Anal. calcd for C21H27N4O6FS-0.15 CF3CO2H: C, 51.20; H, 5.48; N, 11.21; F, 5.51; S, 6.42; found: C, 51.10; H, 5.23; N, 11.21; F, 5.49; S, 6.32.
Example 206




6H-pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, 4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-N-[[2-(tetrahydro-1,1-dioxido-2H-1,2-thiazin-2-yl)phenyl]methyl]-. Off-white solid. 1H NMR (500 MHz, CDCl3) δ ppm: 12.18 (1H, br s), 8.14-8.25 (1H, br), 7.41-7.50 (2H, m), 7.31-7.39 (2H, m), 4.96 (1H, dd, J=14.0, 8.9 Hz), 4.54 (2H, br s), 4.44 (1H, dd, J=14.2, 3.2 Hz), 3.83-3.92 (1H, m), 3.65 (2H, br), 3.38-3.46 (1H, m), 3.18-3.28 (2H, m), 2.32-2.42 (2H, m), 1.89-1.98 (4H, m), 1.55 (6H, d, J=−15.9 Hz); 13C NMR (126 MHz, CDCl3) δ ppm: 168.22, 158.38, 153.54, 147.32, 138.80, 137.00, 130.88, 129.47, 129.29, 127.68, 125.17, 82.60, 60.74, 54.15, 51.05, 39.11, 38.63, 27.76, 27.73, 27.36, 25.04, 24.32. HRMS (ESI) calcd for C22H29N4O6S (M+H): 477.1808; found: 477.1794. Anal. calcd for C22H28N4O6S.0.5CH3CH2OH: C, 55.45; H, 5.92; N, 11.76; S, 6.73; found: C, 55.36; H, 6.11; N, 11.46; S, 6.48.
Example 207




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-diethyl-4-oxo-N-[[2-(tetrahydro-1,1-dioxido-2H-1,2-thiazin-2-yl)phenyl]methyl]. Off-white crystalline solid. 1H NMR (500 MHz, CDCl3) δ ppm: 12.11 (1H, br s), 8.20 (1H, br), 7.42-7.48 (2H, m), 7.33-7.40 (2H, m), 4.93 (1H, dd, J=14.2, 8.4 Hz), 4.45 (1H, dd, J=14.2, 3.8 Hz), 3.94-4.02 (4H, m), 3.87 (1H, ddd, J=13.0, 9.9, 3.7 Hz), 3.39-3.46 (1H, m), 3.15-3.24 (2H, m), 2.29-2.46 (2H, m), 1.79-2.04 (6H, m), 0.81 (3H, t, J=6.7 Hz), 0.78 (3H, t, J=7.3 Hz); 13C NMR (126 MHz, CDCl3) δ ppm: 168.25, 157.91, 151.43, 146.14, 139.14, 136.81, 130.79, 129.46, 129.25, 127.90, 125.96, 81.15, 58.55, 54.22, 51.09, 43.03, 39.57, 31.21, 25.03, 24.34, 7.82, 7.68. HRMS (ESI) calcd for C23H31N4O6S (M+H): 491.1964; found: 491.1953. Anal. calcd for C23H30N4O6S: C, 56.12; H, 6.48; N, 10.91; S, 6.24; found: C, 56.14; H, 6.44; N, 11.07; S, 6.05.
Example 208




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-[2-(methylthio)ethyl]-4-oxo-. A solution of intermediate 141, N-(4-fluorobenzyl)-3-(benzyloxy)-9-(2-(methylthio)ethyl)-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide in 4 mL CF3CO2H was stirred at 60° C. for 1 hr then concentrated. The residue was dissolved in CH2Cl2 and washed with water then concentrated. Trituration with hexanes gave the title compound as a solid. 1H NMR (300 MHz, CDCl3) δ ppm: 12.03 (1H, s) 7.75 (1H, m) 6.84-7.40 (4H, m) 4.39-4.72 (3H, m) 4.04-4.36 (2H, m) 3.68-3.92 (2H, m) 2.49-2.73 (2H, m) 2.07-2.50 (2H, m) 2.02 (3H, s); HRMS (ESI) calcd for C18H20FN3O4S (M+H): 394.1237, found: 394.1234.
Example 209




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-[2-(methylsulfonyl)ethyl]-4-oxo-. A solution of example 208, pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-[2-(methylthio)ethyl]-4-oxo-, (40 mg, 0.11 mmol) in 2 mL CH2Cl2 was treated with excess m-chloroperbenzoic acid (100 mg, 0.4 mmol) and stirred at room temperature for 20 hrs. The reaction mixture was washed with water and concentrated. The crude material was purified by reverse phase HPLC (C18, 12% CH3CN/H2O). The fractions containing the product were concentrated and lyophilized to afford 5 mg (12% yield) of the title compound. 1H NMR (300 MHz, CDCl3) δ ppm: 12.10-12.27 (1H, s) 7.80-8.02 (1H, m) 7.25-7.39 (2H, m) 6.90-7.09 (2H, m) 4.05-4.72 (5H, m) 3.73-3.99 (2H, m) 3.02-3.34 (2H, m) 2.86-2.91 (3H, s) 2.35-2.78 (2H, m); HRMS (ESI) calcd for C18H20FN3O6S (M+H): 426.1135, found: 426.1143.
Example 210




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(3-methyl-5-isoxazolyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 180, N-(4-fluoro-2-(3-methylisoxazol-5-yl)benzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.59 (6H, s, 2×CH3), 2.42 (3H, s, CH3), 4.03 (4H, s, 2×CH2), 4.74 (2H, d, J=6.6 Hz, NCH2), 6.40 (1H, s, aromatic), 7.18 (1H, m, aromatic), 7.32 (1H, dd, J=2.5 Hz and J=9.1 Hz, aromatic), 7.59 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 8.29 (1H, broad t, NH), 11.87 (1H, s, OH). HRMS (ESI+) calculated for C21H22FN4O5 [M+H+]: 429.1574; found: 429.1584.
Example 211




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(2Z)-3-amino-1-oxo-2-butenyl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1HNMR 400 MHz (CDCl3) δ ppm: 1.63 (6H, s, 2×CH3), 2.11 (3H, s, CH3), 4.03 (4H, s, 2×CH2), 4.58 (2H, d, J=6.5 Hz, NCH2), 5.38 (1H, broad, NH), 5.49 (1H, s, CH), 7.09 (1H, m, aromatic), 7.29 (1H, dd, J=3.0 Hz and J=9.1 Hz, aromatic), 7.48 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 9.19 (1H, broad t, NH), 10.26 (1H, broad, NH), 12.21 (1H, s, OH).
Example 212




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(3-bromo-5-isoxazolyl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 181, N-(2-(3-bromoisoxazol-5-yl)-4-fluorobenzyl)-3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (CDCl3) δ ppm: 1.60 (6H, s, 2×CH3), 4.03 (4H, s, 2×CH2), 4.74 (2H, d, J=7.1 Hz, NCH2), 6.63 (1H, s, aromatic), 7.24 (1H, m, aromatic), 7.32 (1H, dd, J=2.5 Hz and J=9.1 Hz, aromatic), 7.63 (1H, dd, J=5.5 Hz and J=8.6 Hz, aromatic), 8.20 (1H, broad t, NH), 11.77 (1H, s, OH).
Example 213




Phosphonic acid, [5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, diethyl ester. Hydrogenolysis of intermediate 187, diethyl 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate (0.089 g, 0.15 mmol) gave 0.065 g (90% yield) of the title compound as a white solid: mp 124° C. 1HNMR 400 MHz (CDCl3) δ ppm: 1.39 (6H, t, J=7.1 Hz, 2×CH3), 1.63 (6H, s, 2×CH3), 4.02 (4H, s, 2×CH2), 4.20 (4H, m, 2×OCH2), 4.78 (2H, d, J=6.6 Hz, NCH2), 7.25 (1H, m, aromatic), 7.49 (1H, m, aromatic), 7.63 (1H, m, aromatic), 9.12 (1H, broad t, NH), 12.1 (1H, broad, OH). HRMS (ESI+) calculated for C21H28FN3O7P [M+H+]: 484.1649; found: 484.1646.
Example 214




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-N-methoxy-9,9-dimethyl-4-oxo-. The title compound can be prepared from intermediate 171, N-(4-fluorobenzyl)-3-(benzyloxy)-N-methoxy-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide. 1HNMR 400 MHz (DMSO-d6) δ ppm: (mixture of rotamers) 1.52 (6H, s, 2×CH3), 3.59 (3H, s, OCH3), 3.89 (2H, broad, CH2), 4.01 (2H, broad, CH2), 4.68 and 4.91 (2H, broad, NCH2), 7.18 (2H, m, aromatics), 7.43 (2H, m, aromatics), 9.88 and 10.2 (1H, broad, OH).
Example 215




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-[[2-(tetrahydro-1,1-dioxido-2H-1,2-thiazin-2-yl)phenyl]methyl]-. The title compound can be prepared from intermediate 101. Pale brown solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 12.11 (1H, s), 8.32-8.29 (1H, m), 7.46-7.42 (2H, m), 7.36-7.32 (2H, m), 4.92 (1H, dd, J=14.3, 8.8 Hz), 4.40 (1H, dd, J=14.1, 3.5 Hz), 3.97 (4H, s), 3.90-3.81 (1H, m), 3.47-3.37 (1H, m), 3.24-3.18 (2H, m), 2.42-2.28 (2H, m), 1.97-1.86 (2H, m), 1.54 (3H, s), 1.50 (3H, s). HRMS [M+H]+ calcd for C21H27N4SO6: 463.1651; found: 463.1669. Anal calcd for C21H26N4SO6: C, 54.53; H, 5.67; N, 12.11; S, 6.93; found: C, 54.53; H, 5.41; N, 12.40; S, 6.69.
Example 216




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-[[2-(1H-1,2,4-triazol-1-yl)phenyl]methyl]-. White solid. 1H-NMR (300 MHz, CDCl3) δ ppm: 11.94 (1H, s), 8.86 (1H, t, J=6.2 Hz), 8.42 (1H, s), 8.16 (1H, s), 7.66 (1H, dd, J=7.3, 1.8 Hz), 7.51-7.42 (2H, m), 7.35-7.32 (1H, m), 4.45 (2H, d, J=6.6 Hz), 3.98 (4H, s), 1.59 (6H, s). HRMS [M+H]+ calcd for C19H21N6O4: 397.1624; found: 397.1609. Anal calcd for C19H20N6O4: C, 57.57; H, 5.08; N, 21.20; found: C, 57.40; H, 4.96; N, 21.09.
Example 217




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-N-(3-phenylpropyl)-1H NMR (500 MHz, DMSO-D6) δ ppm: 1.56 (s, 6H) 1.82-1.89 (m, 2H) 2.62 (t, J=7.48 Hz, 2H) 3.31 (m, 2H) 3.82 (t, J=5.04 Hz, 2H) 3.97 (t, J=5.04 Hz, 2H) 7.16-7.24 (m, 3H) 7.29 (m, 2H) 8.91 (s, 1H) 12.41 (s, 1H). Anal calcd for C19H23N3O4: C, 63.85; H, 6.48; N, 11.75. Found: C, 63.56; H, 6.67; N, 12.01.
Examples 218-259
Examples 218-259, in table 11, were prepared using methods similar to those described for examples 1, 19, 20. The compounds were characterized by LCMS with the observed retention time (RT, minutes) and molecular weight (MS [M+1]) listed in the table.










TABLE 11





Example
Structure
RT
MS


















218




1.55
374.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-(1,3-benzodioxol-5-



ylmethyl)-4,6,7,9-tetrahydro-3-hydroxy-



9,9-dimethyl-4-oxo-


 


219




1.60
330.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-N-



(phenylmethyl)-


 


220




1.63
348.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-



fluorophenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


221




1.99
398.16



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3,4-



dichlorophenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


222




1.61
360.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-N-[(3-methoxyphenyl)methyl]-



9,9-dimethyl-4-oxo-


 


223




1.79
344.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-N-[(3-



methylphenyl)methyl]-4-oxo-


 


224




1.65
366.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,5-



difluorophenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


225




1.27
376.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-N-[(4-hydroxy-3-



methoxyphenyl)methyl]-9,9-dimethyl-4-oxo-


 


226




1.80
364.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-



chlorophenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


227




2.00
398.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,3-



dichlorophenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


228




1.97
398.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,5-



dichlorophenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


229




1.64
390.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,3-



dimethoxyphenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


230




1.64
390.24



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,3-dihydro-1,4-



benzodioxin-6-yl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


231




2.01
378.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-chloro-4-



methylphenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


232




1.75
390.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,4-



dimethoxyphenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


233




1.92
358.2



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,3-



dimethylphenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


234




2.01
398.0



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3,5-



dichlorophenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


235




1.71
390.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,5-



dimethoxyphenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


236




1.98
378.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-chloro-2-



methylphenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


237




1.95
416.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[[3-fluoro-5-



(trifluoromethyl)phenyl]methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


238




1.70
366.0



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,3-



difluorophenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


239




1.71
394.0



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-chloro-4-



methoxyphenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


240




1.80
344.0



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-N-[(4-



methylphenyl)methyl]-4-oxo-


 


241




1.90
416.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[[4-fluoro-3-



(trifluoromethyl)phenyl]methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


242




1.80
360.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-N-[(2-methoxyphenyl)methyl]-



9,9-dimethyl-4-oxo-


 


243




1.90
374.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2-



ethoxyphenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


244




1.60
360.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-N-[(4-methoxyphenyl)methyl]-



9,9-dimethyl-4-oxo-


 


245




1.23
378.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(4-fluoro-3-



methoxyphenyl)methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


246




1.99
416.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[[4-fluoro-2-



(trifluoromethyl)phenyl]methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


247




1.98
376.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-ethyl-4-



fluorophenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


248




1.83
362.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-fluoro-4-



methylphenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


249




1.77
374.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(3-



ethoxyphenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


250




1.59
372.0



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2,3-dihydro-5-



benzofuranyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


251




1.96
378.1



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(5-chloro-2-



methylphenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


252




1.50
396.27



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-N-[[3-(1H-



pyrazol-1-yl)phenyl]methyl]-


 


253




1.27
415.29



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-N-[[4-(4-



morpholinyl)phenyl]methyl]-4-oxo-


 


254




1.12
413.31



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-9,9-dimethyl-4-oxo-N-[[2-(1-



piperidinyl)phenyl]methyl]-


 


255




1.05
345.26



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(2-



aminophenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


256




0.98
345.25



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[(4-



aminophenyl)methyl]-4,6,7,9-tetrahydro-



3-hydroxy-9,9-dimethyl-4-oxo-


 


257




0.98
373.29



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, N-[[4-



(dimethylamino)phenyl]methyl]-4,6,7,9-



tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-


 


258




1.44
346.24



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-N-[(2-hydroxyphenyl)methyl]-



9,9-dimethyl-4-oxo-


 


259




1.22
346.24



Pyrimido[2,1-c][1,4]oxazine-2-



carboxamide, 4,6,7,9-tetrahydro-3-



hydroxy-N-[(4-hydroxyphenyl)methyl]-



9,9-dimethyl-4-oxo-









Examples 260-278
Examples 260-278 were prepared according to the methods used for the preparation of the compounds in table 3 and characterized by LCMS.
Example 260




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(4-morpholinyl)phenyl]methyl]-4-oxo-. LCMS: HPLC retention time=4.03 min, MS=[M+1]415.23.
Example 261




1-piperazinecarboxylic acid, 4-[2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, 1,1-dimethylethyl ester. LCMS: HPLC retention time=4.93 min, MS=[M+1]514.24.
Example 262




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(1-methylethoxy)phenyl]methyl]-4-oxo-. LCMS: HPLC retention time=4.66 min, MS=[M+1]388.21.
Example 263




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[3-(1-methylethoxy)phenyl]methyl]-4-oxo-. LCMS: HPLC retention time 3.97 min., MS=[M+1]4388.21.
Example 264




Benzoic acid, 3-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]-, methyl ester. LCMS: HPLC retention time 3.97 min, MS=[M+1]388.18.
Example 265




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-cyanophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time 3.59 min, MS=[M+1]355.2.
Example 266




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3-chloro-2-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.47 min, MS=[M+1]382.14.
Example 267




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2,3-dihydro-1,4-benzodioxin-5-yl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.15 min, MS=[M+1]388.2.
Example 268




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(3,4-dihydro-2H-1,5-benzodioxepin-6-yl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.12 min, MS=[M+1]402.21.
Example 269




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2,5-dimethylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.64 min, MS=[M+1]358.22
Example 270




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(5-chloro-2-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.44 min, MS=[M+1]382.15.
Example 271




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2,4-dichlorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.88 min, MS=[M+1]398.11.
Example 272




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-chloro-2-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.69 min, MS=[M+1]378.17.
Example 273




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-chloro-6-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.39 min, MS=[M+1]382.13.
Example 274




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(6-chloro-2-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.74 min, MS=[M+1]396.14.
Example 275




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2,6-difluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.49 min, MS=[M+1]380.18.
Example 276




Pyrimido[2,1-c][1, 4]oxazine-2-carboxamide, N-[(2,3-difluoro-4-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.53 min, MS=[M+1]380.18.
Example 277




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(4-chloro-2-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.57 min, MS=[M+1]382.13.
Example 278




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[(2-chloro-6-fluoro-3-methylphenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. LCMS: HPLC retention time=4.68 min, MS=[M+1]396.16.
Example 279




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[(1-methyl-1H-indol-4-yl)methyl]-4-oxo-. 1H NMR (500 MHz, DMSO-d6) δ ppm: 1.56 (s, 6), 3.79 (s, 3), 3.82 (m, 2), 3.97 (m, 2), 4.76 (d, 2), 6.62 (d, 1), 6.96 (d, 1), 7.12 (m, 1), 7.27-7.38 (overlapping m, 2). HRMS [M+1] calcd for C20H23N4O4, 383.1641; found, 383.1717. Anal calcd for C20H22N4O4: C, 62.81; H, 5.79; N, 14.65. Found: C, 62.88; H, 6.08; N, 13.56.
Example 280




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-(2-oxo-1-azetidinyl)phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR (400 MHz, DMSO-d6) δ: 12.17 (1H, s), 9.28 (1H, broad s), 7.37 (1H, dd, J=8.2, 6.7 Hz), 7.23 (1H, dd, J=10.2, 2.4 Hz), 7.04 (1H, m), 4.51 (2H, broad s), 4.35 (2H, broad s), 3.79 (2H, t, J=4.4 Hz), 3.62 (2H, m), 3.11 (2H, t, J=4.4 Hz), 1.81 (2H, m), 1.55 (6H, s). LCMS (M+H)+ m/z 431.
Example 281




Phosphonic acid, [5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, dimethyl ester. Hydrogenation of dimethyl 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate (0.030 g, 0.055 mmol) gave 0.018 g (72%) of the title compound as a white solid 1HNMR 400 MHz (CDCl3) δ (ppm): 1.63 (6H, s, 2×CH3), 3.84 (3H, s, OCH3), 3.87 (3H, s, OCH3), 4.03 (4H, s, 2×CH2), 4.77 (2H, d, J=6.5 Hz, NCH2), 7.28 (1H, m, aromatic), 7.47 (1H, m, aromatic), 7.65 (1H, m, aromatic), 9.0 (1H, broad t, NH) 12.0 (1H, broad, OH). MS (ESI+) m/z 456 [M+H+].
Example 282




Phosphonic acid, [5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, monoethyl ester. The title compound can be prepared from intermediate 188, ethyl hydrogen 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.32 (3H, t, J=7.1 Hz, CH3), 1.60 (6H, s, 2×CH3), 4.02 (4H, s, 2×CH2), 4.12 (2H, m, OCH2), 4.83 (2H, broad, NCH2), 7.18 (1H, m, aromatic), 7.54 (2H, m, aromatics), 8.49 (1H, broad, NH). HRMS (ESI+) calculated for C19H24FN3O7P [M+H+]: 456.1336; found: 456.1353.
Example 283




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(dimethylamino)sulfonyl]-4-fluorophenyl]methyl]-9-ethyl-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. White solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.01 (3H, t, J=7.4 Hz, CH3), 1.9-2.0 (1H, m, CH), 2.15-2.25 (1H, m, CH), 2.92 (6H, s, 2×NCH3), 3.8-3.9 (2H, m, CH2), 4.1-4.3 (2H, m, CH2), 4.45 (1H, m, CH), 4.84 (2H, d, J=7.1 Hz, NCH2), 7.3 (1H, m, aromatic), 7.54 (1H, dd, J=2.8 Hz and J=8.4 Hz, aromatic), 7.71 (1H, dd, J=5.4 Hz and J=8.6 Hz, aromatic), 8.54 (1H, broad t, NH), 11.95 (1H, s, OH). HRMS (ESI+) calculated for C19H24FN4O6S [M+H+]: 455.1401; found: 455.1382.
Example 284




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9-ethyl-N-[[4-fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-e. White crystals; mp 213° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.04 (3H, t, J=7.3 Hz, CH3), 1.9-2.0 (1H, m, CH), 2.25-2.35 (1H, m, CH), 2.56 (3H, s, CH3), 3.8-3.9 (2H, m, CH2), 4.15-4.3 (2H, m, CH2), 4.45-4.52 (3H, m, NCH2 and CH), 7.10 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.20 (1H, m, aromatic), 7.70 (1H, dd, J=6.1 Hz and J=8.6 Hz, aromatic), 8.35 (1H, s, CH), 8.8 (1H, broad t, NH), 12.12 (1H, s, OH). Anal. Calcd for C20H21FN6O4: C, 56.07; H, 4.94; N, 19.61; Found: C, 55.84; H, 4.68; N, 19.47.
Example 285




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9-ethyl-N-[[4-fluoro-2-(5-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-4-oxo-. White solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.06 (3H, t, J=7.3 Hz, CH3), 1.9-2.05 (1H, m, CH), 2.25-2.35 (1H, m, CH), 2.50 (3H, s, CH3), 3.8-3.9 (2H, m, CH2), 4.15-4.45 (4H, m, 2×CH2), 4.47 (1H, m, CH), 7.05 (1H, dd, J=2.5 Hz and J=8.4 Hz, aromatic), 7.27 (1H, m, aromatic), 7.69 (1H, dd, J=5.9 Hz and J=8.6 Hz, aromatic), 8.03 (1H, s, CH), 8.5 (1H, broad t, NH), 11.93 (1H, s, OH). HRMS (ESI+) calculated for C20H22FN6O4 [M+H+]: 429.1687; found: 429.1671.
Example 286




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1-methyl-1H-pyrazol-3-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. White crystals; mp 220° C. (ethyl acetate-ether). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.60 (6H, s, 2×CH3), 4.02 (4H, s, 2×CH2), 4.07 (3H, s, NCH3), 4.68 (2H, d, J=7.0 Hz, NCH2), 6.53 (1H, d, J=2.5 Hz, CH), 7.04 (1H, m, aromatic), 7.28 (1H, dd, J=2.5 Hz and J=9.6 Hz, aromatic), 7.50 (1H, d, J=2.5 Hz, CH), 7.58 (1H, dd, J=6.0 Hz and J=8.6 Hz, aromatic), 9.05 (1H, broad t, NH), 12.44 (1H, s, OH). HRMS (ESI+) calculated for C21H23FN5O4 [M+H+]: 428.1734: found: 428.1732.
Example 287




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1-methyl-1H-pyrazol-5-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo- White crystals; mp 221° C. (ethyl acetate). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.61 (6H, s, 2×CH3), 3.73 (3H, s, NCH3), 4.04 (4H, s, 2×CH2), 4.42 (2H, d, J=6.5 Hz, NCH2), 6.34 (1H, d, J=2.0 Hz, CH), 7.03 (1H, dd, J=2.5 Hz and J=8.6 Hz, aromatic), 7.20 (1H, m, aromatic), 7.56 (1H, dd, J=5.6 Hz and J=8.6 Hz, aromatic), 7.58 (1H, broad t, NH), 7.63 (1H, d, J=2.0 Hz, CH), 11.86 (1H, s, OH). MS (ESI+) m/z 428 [M+H+].
Example 288




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-N-[[2-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4-oxo-e. White crystals; mp 234° C. (dec.) (ethanol). 1HNMR 400 MHz (CDCl3) δ (ppm): 1.64 (6H, s, 2×CH3), 2.58 (3H, s, CH3), 4.03 (4H, s, 2×CH2), 4.52 (2H, d, J=6.8 Hz, NCH2), 7.32-7.38 (H, m, aromatic), 7.44-7.53 (2H, m, aromatics), 7.68-7.75 (1H, m, aromatic), 8.33 (1H, s, CH), 8.81 (1H, broad t, NH), 12.17 (1H, s, OH). Anal. Calcd for C20H22N6O4: C, 58.52; H, 5.40; N, 20.47; Found: C, 58.77; H, 5.50; N, 20.20.
Example 289




Phosphonic acid, [5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-. Hydrogenolysis of benzyl hydrogen 2-((3-(benzyloxy)-9,9-dimethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamido)methyl)-5-fluorophenylphosphonate gave the title acid as a white solid (63% yield). 1H NMR 400 MHz (DMSO-d6) δ (ppm): 1.53 (6H, s, 2×CH3), 3.81 (2H, broad t, CH2), 3.96 (2H, broad t, CH2), 4.74 (2H, d, J=6.1 Hz, NCH2), 7.2-7.5 (3H, m, aromatics), 9.7 (1H, broad, NH), 12.18 (broad, OH). HRMS (ESI+) calculated for C17H20FN3O7P [M+H+]: 428.1023; found: 428.1005.
Example 290




Phosphonic acid, [5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, monomethyl ester. White solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.60 (6H, s, 2×CH3), 3.74 and 3.77 (3H, 2 s, OCH3), 4.02 (4H, s, 2×CH2), 4.80 (2H, broad d, NCH2), 7.2-7.27 (1H, m, aromatic), 7.53-7.64 (2H, m, aromatics), 8.46 (1H, broad t, NH), 11.94 (broad, OH). HRMS (ESI+) calculated for C18H22FN3O7P [M+H+]: 442.1179; found: 442.1177.
Example 291




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(2-oxido-1,3,2-dioxaphospholan-2-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo- White solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.63 (6H, s, 2×CH3), 4.02 (4H, s, 2×CH2), 4.4-4.5 (2H, m, CH2), 4.65-4.72 (2H, m, CH2), 4.80 (2H, d, J=6.5 Hz, NCH2), 7.25-7.42 (2H, m, aromatics), 7.65-7.75 (1H, m, aromatic), 9.07 (1H, broad t, NH), 11.96 (1H, s, OH). HRMS (ESI+) calculated for C19H22FN3O7P [M+H+]: 454.1179; found: 454.1183.
Example 292




Phosphonic acid, [5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, mono(2-hydroxyethyl) ester. White solid 1HNMR 400 MHz (CDCl3) δ (ppm): 1.59 (6H, s, 2×CH3), 3.87 (2H, broad, CH2), 4.02 (4H, s, 2×CH2), 4.18 (2H, broad, CH2), 4.86 (2H, broad d, NCH2), 7.16-7.24 (1H, m, aromatic), 7.48-7.63 (2H, m, aromatics), 8.49 (1H, broad t, NH). HRMS (ESI+) calculated for C19H24FN3O8P [M+H+]: 472.1285; found: 472.1276.
Example 293




Phosphonic acid, [5-fluoro-2-[[[(4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxopyrimido[2,1-c][1,4]oxazin-2-yl)carbonyl]amino]methyl]phenyl]-, bis(2-hydroxyethyl) ester. White solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.62 (6H, s, 2×CH3), 3.85-3.95 (4H, m, 2×CH2), 4.03 (4H, s, 2×CH2), 4.3-4.35 (4H, m, 2×CH2), 4.81 (2H, d, J=6.6 Hz, NCH2), 7.27-7.32 (1H, m, aromatic), 7.5-7.7 (2H, m, aromatics), 8.93 (1H, broad t, NH), 11.97 (1H, s, OH). HRMS (ESI+) calculated for C21H28FN3O9P [M+H+]: 516.1547; found: 516.1525.
Example 294




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(5,5-dimethyl-2-oxido-1,3,2-dioxaphosphorinan-2-yl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo- White solid. 1HNMR 400 MHz (CDCl3) δ (ppm): 1.17 (3H, s, CH3), 1.19 (3H, s, CH3), 1.62 (6H, s, 2×CH3), 4.01 (2H, dd, J=11.1 Hz and J=13.6 Hz, CH2), 4.03 (4H, s, 2×CH2), 4.42 (2H, broad t, J=11 Hz, CH2), 4.83 (2H, d, J=6.6 Hz, NCH2), 7.27-7.33 (1H, m, aromatic), 7.45-7.55 (1H, m, aromatic), 7.65-7.75 (1H, m, aromatic), 8.81 (1H, broad t, NH). HRMS (ESI+) calculated for C22H28FN3O7P [M+H+]: 496.1649; found: 496.1647.
Example 295




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1H-1,2,3-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H 20 NMR 400 MHz (CDCl3) δ (ppm): 8.31 (1H, s), 7.84 (1H, s), 7.58 (1H, m), 7.23 (2H, m), 4.32 (2H, s), 3.90 (2H, m), 3.81 (2H, m), 1.48 (6H, s). HRMS (ESI+) calculated for C19H20FN4O6 [M+H+]415.1530; found: 415.1549.
Example 296




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(5-methyl-1H-1,2,3-triazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dim ethyl-4-oxo-. 1H NMR (400 MHz, CDCl3) δ ppm: 11.82 (1H, s), 8.45 (1H, t, J=6.4 Hz), 7.73 (1H, dd, J=8.7, 5.9 Hz), 7.69 (1H, s), 7.32 (1H, dd, J=8.1, 2.5 Hz), 7.04 (1H, dd, J=8.3, 2.5 Hz), 4.28 (2H, d, J=6.6 Hz), 4.03 (4H, s), 2.32 (3H, s), 1.69 (6H, s), LCMS (ESI, M+H+) m/z 429.
Example 297




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[4-(1,1-dimethylethyl)-1H-1,2,3-triazol-1-yl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (400 MHz, CDCl3) δ ppm: 11.95 (1H, s), 8.82 (1H, t, J=6.6 Hz), 7.72 (1H, dd, J=8.6, 6.1 Hz), 7.64 (1H, s), 7.22 (1H, td, J=8.2, 2.5 Hz), 7.14 (1H, dd, J=8.6, 2.5 Hz), 4.50 (2H, d, J=6.6 Hz), 4.03 (4H, s), 1.66 (6H, s), 1.46 (9H, s), MS (ESI+) m/e 471 [M+H+], HRMS (ESI+) calculated for C23H28FN6O4 [M+H+]471.2156; found: 471.2149.
Example 298




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(4,5-dimethyl-1H-1,2,3-triazol-1-yl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (400 MHz, CDCl3) δ ppm: 11.87 (1H, s), 8.46 (1H, t, J=6.4 Hz), 7.71 (1H, dd, J=8.6, 5.8 Hz), 7.24-7.31 (1H, m), 7.01 (1H, dd, J=8.3, 2.8 Hz), 4.27 (2H, d, J=6.6 Hz), 4.03 (4H, s), 2.41 (3H, s), 2.23 (3H, s), 1.68 (6H, s), LCMS (+ESI, M+H+) m/z 443.
Example 299




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (400 MHz, CDCl3) δ ppm: 11.90 (1H, s), 8.72 (1H, s), 7.92 (1H, s), 7.76 (1H, dd, J=8.6, 6.1 Hz), 7.23-7.28 (1H, m), 7.13 (1H, dd, J=8.3, 2.5 Hz), 4.98 (2H, s), 4.47 (2H, d, J=6.8 Hz), 4.04 (4H, s), 1.67 (6H, s), LCMS (+ESI, M+H+) m/z 445.
Example 300




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-(1H-1,2,3-triazol-1-yl)phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR (400 MHz, CDCl3) δ ppm: 11.94 (1H, s), 8.70 (1H, t, J=6.1 Hz), 7.97 (1H, s), 7.96 (1H, s), 7.78 (1H, dd, J=8.6, 6.1 Hz), 7.22-7.28 (1H, m), 7.14 (1H, dd, J=8.3, 2.5 Hz), 4.58 (2H, s), 4.46 (2H, d, J=6.6 Hz), 3.70 (2H, t, J=6.4 Hz), 1.93-1.99 (2H, m), 1.69 (6H, m), HRMS (ESI+) calculated for C20H22FN6O4 [M+H+]429.1687; found: 429.1695.
Example 301




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[2-[4-(1,1-dimethylethyl)-1H-1,2,3-triazol-1-yl]-4-fluorophenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR 400 MHz (CDCl3) (ppm): 8.73 (1H, s), 7.74 (1H, dd, J=8.6, 6.1 Hz), 7.65 (1H, s), 7.20-7.29 (1H, m), 7.15 (1H, dd, J=8.6, 2.5 Hz), 4.58 (2H, br s,), 4.51 (2H, d, J=6.6 Hz), 3.69 (2H, t, J=6.3 Hz), 1.96 (2H, m), 1.66 (6H, s), 1.46 (9H, s).
Example 302




4H-Pyrimido[1,2-d][1,4]oxazepine-2-carboxamide, N-[[2-(4,5-dimethyl-1H-1,2,3-triazol-1-yl)-4-fluorophenyl]methyl]-6,7,9,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR (400 MHz, CDCl3) δ ppm: 11.87 (1H, s), 8.50 (1H, t, J=6.6 Hz), 7.73 (1H, dd, J=8.7, 5.9 Hz), 7.28 (1H, td, J=8.3, 2.7 Hz), 7.01 (1H, dd, J=8.3, 2.5 Hz), 4.63 (2H, brs), 4.27 (2H, d, J=6.6 Hz), 3.81 (2H, brs), 3.62 (2H, s), 2.40 (3H, s), 2.23 (3H, s), 1.49 (6H, s), LCMS (+ESI, M+H+) m/z 457.
Example 303




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(3-methyl-4H-1,2,4-triazol-4-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. In an open vessel, acetic hydrazine (0.407 g, 5.50 mmol) was dissolved in acetonitrile (2 mL) and dimethylformamide dimethyl acetal (0.732 g, 5.50 mmol) was added. The reaction mixture was warmed to 50° C. for 0.5 h. and then N-(2-amino-4-fluorobenzyl)-3-(benzyloxy)-9,9-diethyl-4-oxo-4,6,7,9-tetrahydropyrimido[2,1-c][1,4]oxazine-2-carboxamide (0.091 g, 0.20 mmol) in acetonitrile (1 mL) was added followed by acetic acid (3 mL). The reaction mixture was heated at 120° C. for 3 h. The solvents were removed in vacuo, and the organic material dissolved in EtOAc (50 mL). The organic solution was washed with H2O, dried (MgSO4) and concentrated in vacuo. The crude material was dissolved in ethyl acetate (10 mL), palladium 10% on charcoal (50 mg) was added and the reaction mixture was stirred under and a hydrogen atmosphere (balloon) for 2 h. Then palladium on charcoal was removed by filtration and the solvent was removed in vacuo. The crude material was purified by crystallization in EtOAc:Hex (4:1) to afford 0.045 g (56%) of the title compound as a white solid. 1H NMR 400 MHz (CDCl3) δ (ppm): 8.40 (1H, s), 7.64-7.75 (1H, m), 7.36 (1H, m), 7.05 (1H, dd, J=7.7, 2.7 Hz), 4.30 (2H, m), 4.04 (4H, s), 2.43 (3H, s), 1.62 (6H, s).
Example 304




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-(2-oxo-3-oxazolidinyl)phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR 400 MHz (MeOD) δ (ppm): 7.55 (1H, dd, J=8.6, 6.3 Hz), 7.25 (1H, dd, J=9.5, 2.7 Hz), 7.14 (1H, td, J=8.4, 2.7 Hz), 4.52-4.63 (6H, m), 4.12 (2H, m), 3.72 (2H, t, J=6.3 Hz), 1.92-1.99 (2H, m), 1.63 (6H, s). HRMS (ESI+) calculated for C21H24FN4O6 [M+H+]447.1680; found: 447.1686.
Example 305




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(ethylsulfonyl)amino]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR 400 MHz (CDCl3) δ (ppm): 11.36 (1H, s), 8.71 (1H, s), 8.14 (1H, m), 7.39 (1H, dd, J=8.6, 6.3 Hz), 7.28 (1H, m), 6.90 (1H, td, J=8.1, 2.5 Hz), 4.60 (2H, d, J=7.1 Hz), 4.04 (4H, s), 3.25 (2H, q, J=7.3 Hz), 1.57 (6H, s), 1.45 (3H, t, J=7.5 Hz).
Example 306




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-(1H-pyrazol-1-yl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. White crystalline powder: HPLC: 2.45 min (AP 98% at 254 nm); LC/MS m/z 414 (M+H); 1H NMR (CDCl3, 500 MHz) δ ppm 1.61 (6H, s, 10,11-CH3), 4.00 (4H, s, 7,8-CH2), 4.46 (2H, d, J=6.7 Hz, 13-CH2), 6.54 (1H, d, J=1.5 Hz, 21-CH), 7.06 (1H, dd, J=9, 2.3 Hz, 16-CH), 7.09 (1H, dt, J=8, 2.5 Hz, 18-CH), 7.61 (1H, dd, J=8.1, 6.3 Hz, 19-CH), 7.77 (2H, d, J=1.2 Hz, 20,21-CH), 9.17 (1H, t, J=6.0 Hz, NH), 12.07 (1H, s, OH); 13C NMR (CDCl3, 125.8 Hz) δ ppm 28.0 (10,11-CH3), 39.6 (13-CH2), 43.1 (7-CH2), 58.3 (8-CH2), 75.9 (9-C), 107.9 (21-CH), 112.0 (d, J=25 Hz, 16-CH), 115.3 (d, J=21 Hz, 18-CH), 126.1 (4-C), 128.2 (d, J=3 Hz, 14-C), 130.5 (20-CH), 134.0 (d, J=8.9 Hz, 19-CH), 140.9 (d, J=11 Hz, 15-C), 141.4 (22-CH), 146.2 (5-C), 151.3 (2-C), 158.0 (6-C), 162.2 (d, J=249 Hz, 17-CF), 167.7 (12-C═O); HRMS (ESI) calcd for C20H21FN5O4 (M+H) 414.1578, found 414.1578 (δ+0.1 ppm); UV (MeOH) λmax 245 nm (ε1.90×104), 306 nm (ε7.82×104); Anal. calcd for C20H20FN5O4: C, 58.10; H, 4.87; N, 16.94; found C, 57.91; H, 4.58; N, 16.85.
Example 307




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(3,5-dimethyl-1H-1,2,4-triazol-1-yl)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. Off-white crystalline powder. HPLC rt=1.69 min. LC/MS m/z 443 (M+H). 1H NMR (CDCl3, 500 MHz) δ ppm 1.61 (6H, s, 10,11-CH3), 2.41 (3H, s, 22-CH3), 2.45 (3H, s, 23-CH3), 3.99 (4H, s, 7,8-CH2), 4.31 (2H, d, J=6.7 Hz, 13-CH2), 6.98 (1H, dd, J=8.4, 2.6 Hz, 16-CH), 7.16-7.23 (1H, dt, J=8.5, 2.5 Hz, 18-CH), 7.65 (1H, dd, J=8.7, 6.0 Hz, 19-CH), 8.40 (1H, t, J=6.4 Hz, NH), 11.99 (1H, s, OH). 13C NMR (CDCl3, 125.8 Hz) δ ppm 12.7 (22-CH3), 14.0 (23-CH3), 28.1 (10,11-CH3), 38.7 (13-CH2), 43.1 (7-CH2), 58.2 (8-CH2), 75.9 (9-C), 114.2 (d, J=24 Hz, 16-CH), 117.3 (d, J=21 Hz, 18-CH), 125.6 (4-C), 130.8 (d, J=3.8 Hz, 14-C), 133.3 (d, J=8.8 Hz, 19-CH), 137.1 (d, J=9.6 Hz, 15-C), 146.4 (5-C), 151.6 (2-C), 153.9 (20-C), 158.0 (6-C), 160.7 (21-C), 161.9 (d, J=251 Hz, 17-CF), 168.1 (12-C═O). HRMS (ESI) calcd for C21H24FN6O4 (M+H) 443.1843, found 443.1826 (δ-3.9 ppm). UV (MeOH) λmax 316 nm (ε7.61×103). Anal. calcd for C21H23FN6O4.0.3 EtOH: C56.86; H5.48; N18.42; found C, 56.49; H, 5.12; N, 18.68.
Example 308




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[5-(hydroxymethyl)-1H-1,2,4-triazol-1-yl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. (white crystals, CH3CN). HPLC rt=1.63 min. LC/MS m/z 445 (M+H). 1H NMR (CDCl3, 500 MHz) δ ppm 1.61 (6H, s, 10,11-CH3), 2.42 (3H, br. OH?), 4.00 (4H, s, 7,8-CH2), 4.32 (2H, d, J=6.4 Hz, 13-CH2), 4.73 (2H, s, 22-CH2), 7.21 (1H, dd, J=8.3, 2.5 Hz, 16-CH), 7.27 (1H, m, 18-CH), 7.68 (1H, dd, J=8.5, 5.8 Hz, 19-CH), 8.11 (1H, s, 21-CH), 8.53 (1H, t, J=6.5 Hz, NH),  ̃11.9 (1H, br, OH). 13C NMR (CDCl3, 125.8 Hz) δ ppm 28.0 (10,11-CH3), 38.5 (13-NCH2), 43.1 (7-NCH2), 55.0 (22-OCH2), 58.3 (8-OCH2), 75.9 (9-C), 114.7 (d, J=25 Hz, 16-CH), 118.1 (d, J=20 Hz, 18-CH), 125.6 (4-C), 130.9 (d, J=3.9 Hz, 14-C), 133.4 (d, J=9.6 Hz, 19-CH), 136.4 (d, J=9.6 Hz, 15-C), 146.3 (5-C), 150.9 (21-CH), 151.7 (2-C), 156.0 (20-C), 157.9 (6-C), 162.0 (d, J=252 Hz, 17-CF), 168.0 (12-C═O). HRMS (ESI) calcd for C20H22FN6O5 (M+H) 445.1636, found 445.1630. UV (MeOH) λmax 316 nm (ε8.06×103). Anal. calcd for C20H21FN6O5.0.3H2O: C53.40; H4.84; N18.68; found C, 53.15; H, 4.61; N, 18.44. The amorphous powder was crystallized from acetonitrile to obtain an acetonitrile solvate as white crystals: HPLC rt=1.65 min. LC/MS m/z 445 (M+H). 1H NMR (CDCl3, 500 MHz) δ ppm 1.61 (6H, s, 10,11-CH3), 2.00 (3H, s, CH3CN), 4.00 (4H, s, 7,8-CH2), 4.32 (2H, d, J=6.4 Hz, 13-CH2), 4.73 (2H, s, 22-CH2), 7.21 (1H, dd, J=8.3, 2.5 Hz, 16-CH), 7.27 (1H, m, 18-CH), 7.68 (1H, dd, J=8.5, 5.8 Hz, 19-CH), 8.11 (1H, s, 21-CH), 8.53 (1H, t, J=6.5 Hz, NH),  ̃11.9 (1H, br, OH). 13C NMR (CDCl3, 125.8 Hz) δ ppm 1.97 (CH3CN), 28.0 (10,11-CH3), 38.5 (13-NCH2), 43.1 (7-NCH2), 55.0 (22-OCH2), 58.3 (8-OCH2), 75.9 (9-C), 114.7 (d, J=25 Hz, 16-CH), 116.4 (CH3CN), 118.1 (d, J=20 Hz, 18-CH), 125.6 (4-C), 130.9 (d, J=3.9 Hz, 14-C), 133.4 (d, J=9.6 Hz, 19-CH), 136.4 (d, J=9.6 Hz, 15-C), 146.3 (5-C), 150.9 (21-CH), 151.7 (2-C), 156.0 (20-C), 157.9 (6-C), 162.0 (d, J=252 Hz, 17-CF), 168.0 (12-C═O).
Example 309




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[[2-(acetyloxy)ethyl]thio]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (300 MHz, CHLOROFORM-D) δ ppm 1.55 (s, 6H) 1.99 (s, 3H) 3.04-3.28 (m, 2H) 3.98 (s, 4H) 4.12-4.33 (m, 2H) 4.47-4.73 (m, 2H) 6.81-6.99 (m, 1H) 7.03-7.43 (m, 2H) 7.90-8.10 (m, 1H) 11.85 (s, 1H); HRMS calcd. for C21H24FN3O6S (M+H) 466.1448, found 466.1439.
Example 310




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[[2-(acetyloxy)ethyl]sulfonyl]-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (300 MHz, CHLOROFORM-D) δ ppm 1.55 (s, 6H) 1.81 (s, 3H) 3.57 (t, J=6.04 Hz, 2H) 3.97 (s, 4H) 4.44 (t, J=6.04 Hz, 2H) 4.77 (d, J=6.95 Hz, 2H) 7.27-7.42 (m, 1H) 7.58-7.79 (m, 2H) 8.52 (t, J=6.77 Hz, 1H) 11.69 (s, 1H)); HRMS calcd. for C21H24FN3O8S (M+H) 498.1346, found 498.1350.
Example 311




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-(ethylthio)-4-fluorophenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (300 MHz, CHLOROFORM-D) δ ppm 1.33 (t, J=7.32 Hz, 3H) 1.55 (s, 6H) 2.97 (q, J=7.32 Hz, 2H) 3.98 (s, 4H) 4.56-4.63 (d, J=6.78 Hz, 2H) 6.70-7.09 (m, 2H) 7.31 (dd, J=8.42, 5.86 Hz, 1H) 8.00-8.22 (t, J=6.78 Hz, 1H) 11.90 (s, 1H)); HRMS calcd. for C19H22FN3O4S (M+H) 408.1393, found 408.1385.
Example 312




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(2-hydroxyethyl)sulfonyl]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (300 MHz, CHLOROFORM-D) δ ppm 1.56 (s, 6H) 3.25-3.58 (m, 2H) 3.97 (s, 4H) 4.02-4.16 (m, 2H) 4.79 (d, J=6.59 Hz, 2H) 7.25-7.40 (m, 1H) 7.58-7.82 (m, 2H) 8.42-8.62 (m, 1H) 11.67 (s, 1H)); HRMS calcd. for C19H22FN3O7S (M+H) 456.1241, found 456.1224.
Example 313




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[4-fluoro-2-[(2-hydroxyethyl)thio]phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-. 1H NMR (300 MHz, CHLOROFORM-D) δ ppm 1.56 (s, 6H) 2.02 (t, J=5.86 Hz, 1H) 3.06-3.21 (m, 2H) 3.79 (q, J=6.10 Hz, 2H) 3.98 (s, 4H) 4.66 (d, J=6.22 Hz, 2H) 6.82-7.17 (m, 2H) 7.34 (dd, J=8.60, 6.04 Hz, 1H) 7.99-8.17 (m, 1H) 11.86 (s, 1H)); HRMS calcd. for C19H22FN3O5S (M+H) 424.1342, found 423.1332.
Example 314




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, 9-ethyl-N-[[4-fluoro-2-(methylsulfonyl)phenyl]methyl]-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. 1H NMR (500 MHz, CDCl3) δ ppm 11.72 (1H, s), 8.57 (1H, t, J=6.6 Hz), 7.75 (1H, dd, J=8.2, 2.7 Hz), 7.70 (1H, dd, J=8.5, 5.2 Hz), 7.34 (1H, dt, J=8.0, 2.6 Hz), 4.76-4.87 (2H, m), 4.12 (1H, dt, J=13.9, 3.5, 3.4 Hz), 4.02 (1H, dt, J=−12.4, 4.0 Hz), 3.88-3.96 (1H, m), 3.80-3.87 (1H, m), 3.17 (3H, s), 1.99-2.08 (1H, m), 1.79-1.88 (1H, m), 1.53 (3H, s), 0.80 (3H, t, J=7.3 Hz); 13C NMR (126 MHz, CDCl3) δ ppm 168.17, 163.12, 161.11, 157.82, 151.83, 146.09, 140.74, 140.69, 135.13, 135.07, 132.80, 132.77 125.63, 121.63, 121.46, 117.63, 117.44, 78.63, 58.31, 45.11, 43.18, 40.31, 34.39, 25.38, 7.79. HRMS (ESI) calcd for C19H23N3O6FS (M+H) 440.1292, found 440.1301. Elem. Anal. calcd for C19H22N3O6FS: C, 51.93; H, 5.13; N, 9.46; S, 7.22; F, 4.28; found: C, 51.78; H, 4.80; N, 9.39; S, 7.19; F, 4.30. HPLC rt=2.27 min. Off-white solid.
Example 315




Pyrimido[2,1-c][1,4]oxazine-2-carboxamide, N-[[2-[(dimethylamino)sulfonyl]-4-fluorophenyl]methyl]-9-ethyl-4,6,7,9-tetrahydro-3-hydroxy-9-methyl-4-oxo-. 1H NMR (500 MHz, CDCl3) δ ppm 11.83 (1H, s), 8.61 (1H, t, J=6.4 Hz), 7.68 (1H, dd, J=8.5, 5.2 Hz), 7.50 (1H, dd, J=8.4, 2.6 Hz), 7.25-7.29 (1H, m), 4.80 (2H, ddd, J=18.5, 14.4, 7.0 Hz), 4.12 (1H, dt, J=13.8, 3.5 Hz), 4.02 (1H, dt, J=12.4, 3.9 Hz), 3.92 (1H, ddd, J=12.3, 8.9, 3.2 Hz), 3.84 (1H, ddd, J=113.7, 9.1, 4.1 Hz), 2.90 (6H, s), 2.01-2.08 (1H, m), 1.80-1.87 (1H, m), 1.53 (3H, s), 0.81 (3H, t, J=7.3 Hz); 13C NMR (126 MHz, CDCl3) δ ppm 168.03, 162.70, 160.69, 157.89, 151.63, 146.05, 138.56, 138.52, 135.17, 135.11, 132.65, 132.62, 125.81, 120.37, 120.21, 116.92, 116.73, 78.67, 58.34, 43.16, 40.28, 37.58, 34.39, 25.35, 7.80. HRMS (ESI) calcd for C20H26N4O6FS (M+H) 469.1557, found 469.1554. HPLC rt=2.52 min. Off-white solid.
Example 316




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-(5-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR (500 MHz, CDCl3) δ ppm 11.88 (1H, br), 8.51 (1H, t, J=6.10 Hz), 8.02 (1H, s), 7.68 (1H, dd, J=8.70, 5.95 Hz), 7.22-7.27 (1H, m), 7.02 (1H, dd, J=8.24, 2.44 Hz), 4.55 (2H, br), 4.29 (2H, d, J=6.41 Hz), 3.68 (2H, t, J=6.56 Hz), 2.50 (3H, s), 1.90-1.97 (2H, m), 1.64 (6H, s); 13C NMR (126 MHz, CDCl3) δ ppm 168.00, 163.00, 161.01, 158.24, 153.64, 153.50, 150.58, 147.36, 133.63, 133.55, 130.81, 130.78, 124.85, 117.92, 117.75, 114.51, 114.32, 82.47, 60.89, 38.73, 38.63, 27.76, 27.37, 12.63. HRMS (ESI) calcd for C21H24N6O4F (M+H) 443.1843, found 443.1826. HPLC rt=1.94 min. White crystalline solid.
Example 317




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[4-fluoro-2-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR (500 MHz, CDCl3) δ ppm 12.08 (1H, br), 8.58-8.68 (1H, m), 8.31 (1H, s), 7.66-7.73 (1H, m), 7.14-7.21 (1H, m), 7.05-7.11 (1H, m), 4.55 (2H, br), 4.47 (2H, d, J=6.71 Hz), 3.64-3.72 (2H, m), 2.52 (3H, s), 1.89-1.98 (2H, m), 1.62 (6H, s). HRMS (ESI) calcd for C21H24N6O4F (M+H) 443.1843, found 443.1856. HPLC rt=2.04 min. Off-white solid.
Example 318




Phosphonic acid, [5-fluoro-2-[[[(4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-6H-pyrimido[2,1-c][1,4]oxazepin-2-yl)carbonyl]amino]methyl]phenyl]-, dimethyl ester. 1H NMR (500 MHz, CDCl3) δ ppm 12.02-12.11 (1H, br), 8.94 (1H, t, J=6.71 Hz), 7.59-7.65 (1H, m), 7.40-7.47 (1H, m), 7.21-7.26 (1H, m), 4.74 (2H, d, J=6.71 Hz), 4.49-4.59 (2H, br), 3.81-3.83 (3H, s), 3.79-3.81 (3H, s), 3.63-3.68 (2H, m), 1.89-1.96 (2H, m), 1.59-1.64 (6H, s); 13C NMR (126 MHz, CDCl3) δ ppm 168.01, 158.34, 153.28, 147.32, 137.64, 134.25, 134.19, 134.12, 134.06, 125.20, 120.43, 120.36, 120.25, 120.18, 120.08, 82.58, 60.79, 53.26, 53.22, 41.45, 41.42, 38.53, 27.75, 27.37. HRMS (ESI) calcd for C20H26N3O7FP (M+H) 470.1492, found 470.1487. Elem. Anal. calcd for C20H25N3O7FP: C, 51.17; H, 5.36; N, 8.95; found: C, 51.21; H, 5.24; N, 8.70. HPLC rt=2.09 min. White crystalline solid.
Example 319




6H-Pyrimido[2,1-c][1,4]oxazepine-2-carboxamide, N-[[2-(3,5-dimethyl-1H-1,2,4-triazol-1-yl)-4-fluorophenyl]methyl]-4,7,8,10-tetrahydro-3-hydroxy-10,10-dimethyl-4-oxo-. 1H NMR (500 MHz, CHLOROFORM-D) δ ppm 12.01 (1H, s), 8.31 (1H, t, J=6.26 Hz), 7.66 (1H, dd, J=8.55, 6.10 Hz), 7.21 (1H, td, J=8.24, 2.44 Hz), 6.99 (1H, dd, J=8.55, 2.44 Hz), 4.55 (2H, s), 4.32 (2H, d, J=6.71 Hz), 3.67 (2H, t, J=6.41 Hz), 2.44 (3H, s), 2.42 (3H, s), 1.90-1.98 (2H, m), 1.62 (6H, s); 13C NMR (126 MHz, CHLOROFORM-D) δ ppm 168.11, 162.95, 160.95, 160.59, 158.24, 153.78, 153.45, 147.48, 137.07, 136.99, 133.42, 133.35, 130.80, 124.86, 117.45, 117.29, 114.32, 114.12, 82.42, 60.83, 38.65, 27.90, 27.34, 13.94, 12.71. HRMS (ESI) calcd for C22H26N6O4F (M+H) 457.2000, found 457.2018. Elem. Anal. calcd for C22H25N6O4F: C, 57.54; H, 5.55; N, 18.30; found: C, 57.91; H, 5.35; N, 18.05. HPLC rt 1.82 min. White solid.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide (N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide, see structure below). Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide which is substantially pure.




Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide designated as form N-1. Another aspect of the invention is the crystalline form N-1 which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a powder x-ray diffraction pattern comprising 20 values selected from 7.0±0.1, 11.9±0.1, 17.0±0.1, 18.8±0.1, and 23.9±0.1. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a powder X-ray diffraction pattern substantially the same as that of FIG. 1. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a differential scanning calorimetry thermogram substantially the same as that of FIG. 2. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a thermal gravimetric analysis scan substantially the same as that of FIG. 3. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a SSNMR spectra substantially the same as that of FIG. 4. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a moisture sorption isotherm substantially the same as that of FIG. 5. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide designated as form N-2. Another aspect of the invention is the crystalline form N-2 which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a powder x-ray diffraction pattern comprising 20 values selected from 8.6±0.1, 10.9±0.1, 15.2±0.1, 15.9±0.1, 19.6±0.1, 21.9±0.1, and 25.8±0.1. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a powder X-ray diffraction pattern substantially the same as that of FIG. 6. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a differential scanning calorimetry thermogram substantially the same as that of FIG. 7. Another aspect of the invention is the form which is substantially pure.
Another aspect of the invention is a crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide characterized by a thermal gravimetric analysis scan substantially the same as that of FIG. 8. Another aspect of the invention is the form which is substantially pure.
“Substantially” means materially equivalent as determined by those skilled in the art. “Substantially pure” means the crystalline form comprises at least about 90 weight % of the compound. The remaining material may comprise other forms or impurities. “Substantially free” means that the compound comprises less than about weight 10% of other forms or impurities.
Single crystal data: method 1. Data can be collected on a Bruker-Nonius CAD4 serial diffractometer. Unit cell parameters can be obtained through least-squares analysis of the experimental diffractometer settings of 25 high-angle reflections. Intensities can be measured using Cu Kα radiation (λ=1.5418 Å) at a constant temperature with the θ-2θ variable scan technique and were usually corrected only for Lorentz-polarization factors. Background counts can be collected at the extremes of the scan for half of the time of the scan. Alternately, single crystal data can be collected on a Bruker-Nonius Kappa CCD 2000 system using Cu Kα radiation (λ=1.5418 Å). Indexing and processing of the measured intensity data can be carried out with the HKL2000 software package in the Collect program suite. When indicated, crystals are usually cooled in the cold stream of an Oxford cryo system during data collection. The structures can be solved by direct methods and refined on the basis of observed reflections using either the SDP software package with minor local modifications or the crystallographic package, MAXUS.
The derived atomic parameters (coordinates and temperature factors) were usually refined through full matrix least-squares. The function minimized in the refinements was ΣW(|FO|−|FC|)2. R is defined as Σ||FO|−|FC||/Σ|FO| while RW=[ΣW(|FO|−|FC|)2/ΣW|FO|2]1/2 where w is an appropriate weighting function based on errors in the observed intensities. Difference maps were usually examined at all stages of refinement. Hydrogens are introduced in idealized positions with isotropic temperature factors, but no hydrogen parameters are varied.
Single crystal data: method 2. Alternatively, a Bruker SMART 2K CCD diffractometer equipped with graphite-monochromated Cu Kα radiation, (λ=1.54056 Å) can be used to collect diffraction data at room temperature. A full data set can be collected using the ω scan mode over the 2θ range with a crystal-to-detector distance of 4.98 cm. An empirical absorption correction utilized the SADABS routine associated with the diffractometer (Bruker AXS. 1998, SMART and SAINTPLUS. Area Detector Control and Integration Software, Bruker AXS, Madison, Wis., USA). The final unit cell parameters are usually determined using the entire data set.
Structures can be solved by direct methods and refined by the full-matrix least-squares techniques, using the SHELXTL software package (Sheldrick, G M. 1997, SHELXTL. Structure Determination Programs. Version 5.10, Bruker AXS, Madison, Wis., USA.). The function minimized in the refinements was ΣW(|FO|−|FC|)2. R is defined as Σ||FO|−|FC||/Σ|FO| while RW=[ΣW(|FO|−|FC|)2/ΣW|FO|2]1/2, where w is an appropriate weighting function based on errors in the observed intensities. Difference Fourier maps are examined at all stages of refinement. All non-hydrogen atoms are refined with anisotropic thermal displacement parameters. The hydrogen atoms associated with hydrogen bonding are located in the final difference Fourier maps while the positions of the other hydrogen atoms are calculated from an idealized geometry with standard bond lengths and angles. They are assigned isotropic temperature factors and included in structure factor calculations with fixed parameters.
PXRD (Philips). About 200 mg are pack by the backloading method into a Philips powder X-ray diffraction (PXRD) sample holder. The sample is transferred to a Philips MPD unit (45 KV, 40 mA, Cu Kα). Data are collected at room temperature in the 2 to 32 2-theta range (continuous scanning mode, scanning rate 0.03 degrees/sec., auto divergence and anti scatter slits, receiving slit: 0.2 mm, sample spinner: ON).
PXRD (GADDS: method 1). X-ray powder diffraction (PXRD) data are obtained using a Bruker C2 GADDS. The radiation is Cu Kα (40 KV, 50 mA). The sample-detector distance is 15 cm. Powder samples are placed in sealed glass capillaries of 1 mm or less in diameter; the capillary is rotated during data collection. Data is collected for 3≦2θ≦35° with a sample exposure time of at least 2000 seconds. The resulting two-dimensional diffraction arcs are integrated to create a traditional 1-dimensional PXRD pattern with a step size of 0.02 degrees 2θ in the range of 3 to 35 degrees 2θ.
PXRD (GADDS: method 2). X-ray powder diffraction (PXRD) data is obtained using a Bruker GADDS manual chi platform goniometer. Powder samples are placed in thin walled glass capillaries of 1 mm or less in diameter; the capillary is rotated during data collection. The sample-detector distance was usually 17 cm. The radiation was Cu Kα(λ=1.5418 Ang). Data is collected for 3≦2θ≦35° with a sample exposure time of at least 300 seconds.
PXRD (Bruker D8 Advance). The powder diffraction data are obtained with a Bruker D8 Advance diffractometer (Karlsruhe, Germany) which was equipped with a monochromated CuKα source operating at a tube load of 40 KV and 40 mA. A system of divergence, anti-scatter and receiving slits of 1.0, 1.0 and 0.1 mm, respectively, was usually employed. The sample was usually scanned in a locked coupled scan mode (step size 0.05°; scan speed 0.4 s per step) from 5 to 40° 2θ.
Hybrid PXRD (from low temperature). “Hybrid” simulated powder X-ray patterns can be generated as described in the literature (Yin. S.; Scaringe, R. P.; DiMarco, J.; Galella, M. and Gougoutas, J. Z., American Pharmaceutical Review, 2003, 6,2, 80). The room temperature cell parameters are obtained by performing a cell refinement using the CellRefine.xls program. Input to the program includes the 2-theta position of ca. 10 reflections, obtained from the experimental room temperature powder pattern; the corresponding Miller indices, hkl, are assigned based on the single-crystal data collected at low temperature. A new (hybrid) PXRD can be calculated (by either of the software programs, Alex or LatticeView) by inserting the molecular structure determined at low temperature into the room temperature cell obtained in the first step of the procedure. The molecules are inserted in a manner that retains the size and shape of the molecule and the position of the molecules with respect to the cell origin, but, allows intermolecular distances to expand with the cell.
Hybrid PXRD (from isostructural analog). “Hybrid” simulated powder X-ray patterns can be generated as described in the literature (Yin. S.; Scaringe, R. P.; DiMarco, J.; Galella, M. and Gougoutas, J. Z., American Pharmaceutical Review, 2003, 6,2, 80). The room temperature cell parameters are obtained by performing a cell refinement using the CellRefine.xls program. Input to the program includes the 2-theta position of ca. 10 reflections, obtained from the experimental room temperature powder pattern; the corresponding Miller indices, hkl, are assigned based on the single-crystal data collected for an isostructural analog. A crystal structure for the molecule of interest can be generated in a two step process: (1) by replacing the analog molecule in the experimental analog crystal structure with the molecule of interest. This step fixes the orientation and position of the molecule of interest in the unit cell of the analog compound; (2) Inserting the molecule of interest into the room temperature cell obtained from the experimental PXRD of the molecule of interest, as described above. In this step, the molecules are inserted in a manner that retains the size and shape of the molecule and the position of the molecules with respect to the cell origin, but, allows intermolecular distances to expand/contract with the cell. A new (hybrid) PXRD can be calculated (by either of the software programs, Alex or LatticeView) based on the crystal structure generated as described above.
DSC (open pan). Differential scanning calorimetry (DSC) experiments can be performed in a TA InstrumentsTM model Q1000 or 2920. The sample (about 2-6 mg) is weighed in an aluminum pan and recorded accurately recorded to a hundredth of a milligram, and transferred to the DSC. The instrument is purged with nitrogen gas at 50 mL/min. Data are collected between room temperature and 300° C. at 10° C./min heating rate. The plot is usually made with the endothermic peaks pointing down.
TGA (open pan). Thermal gravimetric analysis (TGA) experiments can be performed in a TA InstrumentsTM model Q500 or 2950. The sample (about 10-30 mg) is placed in a platinum pan previously tared. The weight of the sample is measured accurately and recorded to a thousand of a milligram by the instrument The furnace is purged with nitrogen gas at 100 mL/min. Data is collected between room temperature and 300° C. at 10° C./min heating rate.
DSC (sealed pan). Differential scanning calorimetry (DSC) experiments can be performed in a TA InstrumentsTM model Q1000 or 2920. The sample (about 2-6 mg) is weighed in an pinpricked hermetically sealed aluminum pan and recorded accurately recorded to a hundredth of a milligram, and transferred to the DSC. The instrument is purged with nitrogen gas at 50 mL/min. Data is collected between room temperature and 300° C. at 10° C./min heating rate. The plot was usually made with the endothermic peaks pointing down.
TGA (sealed pan). Thermal gravimetric analysis (TGA) experiments can be performed in a TA InstrumentsTM model Q500 or 2950. The sample (about 10-30 mg) is placed in a pinpricked hermetically sealed aluminum pan on a platinum pan, both previously tared. The weight of the sample is measured accurately and recorded to a thousand of a milligram by the instrument. The furnace is purged with nitrogen gas at 100 mL/min. Data are collected between room temperature and 300° C. at 10° C./min heating rate.
TG/MS. Thermal gravimetry-mass spectra (TG-MS) data can be collected using TGA/SDTA 851e (US Mettler Toledo Inc. Columbus, Ohio) coupled with ThermostarTM Quadruple Mass Spectrometer (Pfeiffer-Vacuum Inc, Nashua, N.H.). A fused silica capillary that is maintained at 200° C. was employed as the interface. He gas at the rate of 30 ml/min is used as carrying gas. In a typical experiment, the sample ( ̃5 mg) is kept isotherm for 5 min at 25° C., then it is heated from 25 to 300 C at heating rate of 10 C/min. The evolved gas from the TG experiment is directed simultaneously to the MS Spectrometer. The measurement is performed in MID (Multiple Ion Detection) mode.
Solid-State Nuclear Magnetic Resonance (SSNMR). All solid-state C-13 NMR measurements can be made with a Bruker DSX-400, 400 MHz NMR spectrometer. High resolution spectra are obtained using high-power proton decoupling and the TPPM pulse sequence and ramp amplitude cross-polarization (RAMP-CP) with magic-angle spinning (MAS) at approximately 12 kHz (A. E. Bennett et al, J. Chem. Phys., 1995, 103, 6951), (G. Metz, X. Wu and S. O, Smith, J. Magn. Reson. A, 1994, 110, 219-227). Approximately 70 mg of sample, packed into a canister-design zirconia rotor is used for each experiment. Chemical shifts (δ) are referenced to external adamantane with the high frequency resonance being set to 38.56 ppm (W. L. Earl and D. L. VanderHart, J. Magn. Reson., 1982, 48, 35-54).
Raman spectra can be acquired at a resolution of 4 cm−1 with 128 scans co-added, using a Nicolet 950 FT-Raman spectrophotometer. The wavelength of the laser excitation was usually 1064 nm. A CaF2 beam splitter and a high sensitivity InGaS detector are usually used.
IR. Infra-red spectra can be acquired at a resolution of 4 cm−1 with 32 scans co-added, using a Nicolet 560 FT-IR Spectrophotometer, incorporating a KBr beam-splitter and DTGS detector. Sample preparation is via the attenuated total reflectance method (ATR) using a single-bounce diamond ATR sampling accessory (DurasamplIR) from SensIR. An ATR correction step is usually included to correct the pathlength.
VTI (dry on). Moisture sorption isotherms can be collected in a VTI SGA-100 Symmetric Vapor Analyzer using approximately 10 mg of sample. The sample is dried at 60° C. until the loss rate of 0.0005 wt %/min was obtained for 10 minutes. The sample is tested at 25° C. and 3 or 4, 5, 15, 25, 35, 45, 50, 65, 75, 85, and 95% RH. Equilibration at each RH is reached when the rate of 0.0003 wt %/min for 35 minutes was achieved or a maximum of 600 minutes.
VTI (dry off). Moisture sorption isotherms can be collected in a VTI SGA-100 Symmetric Vapor Analyzer using approximately 10 mg of sample. The sample is tested at 25° C. and 3, 5, 25, 35, 45, 50, 65, 75, 85, and 95% RH. Equilibration at each RH is reached when the rate of 0.0003 wt %/min for 35 minutes was achieved or a maximum of 600 minutes.
Hot stage: method 1. Crystals are placed on a glass slide, covered with a cover slip, and heated on a Linkam LTS350 hot stage mounted on a microscope. The heating rate is controlled at 10°/min for the temperature range, ambient to 300° C. The crystals are observed visually for evidence of phase transformation, changes in birefringence, opacity, and melting etc.
Crystals are placed on a glass slide, and heated on a Linkam LTS350 hot stage mounted on a microscope. The heating rate is controlled at 10°/min for the temperature range, ambient to 308° C. The crystals are observed visually for evidence of phase transformation, changes in birefringence, opacity, and melting etc.
Hot stage: method 2. Data can be collected on a Mettler FP 82 HT Hot Stage or FP 84HT TA Microscopic Cell mounted on a microscope, using 70× and 200× nominal magnification and various filters. The heating rate is controlled at 10° C./min for the temperature range, ambient to 305° C. The crystals are observed visually for evidence of phase transformation, changes in birefringence, opacity, and melting etc.
Example 320
Crystalline form N-1 can be made by recrystallizing N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido[2,1-c][1,4]oxazine-2-carboxamide in ethyl acetate at 55° C. (where solution is observed) and allowing the solution to cool to room temperature (where a slurry is observed). The slurry is stirred and can be monitored until conversion to form N-1 is complete. The solid is then filtered and dried.
Example 321
Crystalline form N-1 can be recrystallized in ethyl acetate/hexane. Single-crystal analysis characterized form N-1 as colorless plates.
Example 322
Form N-2 can be made according to the following studies. A slurry conversion study was done at various temperatures to evaluate the thermodynamic relationship between N-1 and N-2. In this experiment, slurries of N-1 in various solvents (for example, acetone or ethyl acetate) at different temperatures were left to saturate for 4 hours. A 50:50 mixture of N-1:N-2 was then added and PXRD collected on the slurry (t=0), in all cases this initial pattern showed the presence of both N-1/N-2 polymorphs. The mixtures were left at various temperatures and slurry PXRD recollected at time intervals. After at least four days all the slurries had fully converted over to either N-1 or N-2.
Three temperatures were investigated. The study at room temperature all converted over to N-1 after four days. At 100° C. the mixture fully converted over to N-1 after a day. At 130° C. the PXRD pattern showed the mixture had fully converted to N-2 after eight hours. Experimentally, N-1 converted to N-2 between 100° C. and 130° C.
Form N-2 can be crystallized from the melt of form N-1. Single-crystal analysis characterized form N-2 as colorless blocks.
Example 323
Characteristic diffraction peak positions (degrees 2θ±0.1)@ RT, based on a high quality pattern collected with a diffractometer (CuKα) with a spinning capillary with 2θ calibrated with a NIST other suitable standard.










TABLE 2







N-1
N-2



















7.0
8.6



11.9
10.9



17.0
15.2



18.8
15.9



23.9
19.6




21.9




25.8










Example 324










Crystal data for form N-1.


















Temperature
293(2) K



Wavelength
0.71073 Å



Crystal system, space group
Orthorhombic, Pbca



Unit cell dimensions
a = 9.2120(3)Å α = 90°




b = 14.4866(4)Å β = 90°




c = 25.1791(11)Å γ = 90°



Volume
3360.2(2)Å3



Z, Calculated density
8, 1.373 Mg/m3



Absorption coefficient
0.106 mm−1



F(000)
1456



Crystal size
0.24 × 0.18 × 0.03 mm



Theta range for data collection
2.93 to 25.03°



Limiting indices
−10 <= h <= 9




−17 <= k <= 15




−28 <= l <= 29



Reflections collected/unique
15242/2953 [R(int) = 0.0615]



Completeness to theta = 25.03
99.7%



Absorption correction
None



Refinement method
Full-matrix least-squares on




F2



Data/restraints/parameters
2953/0/235



Goodness-of-fit on F2
1.022



Final R indices [I > 2[](I)]
R1 = 0.0446, wR2 = 0.1054



R indices (all data)
R1 = 0.0816, wR2 = 0.1214



Extinction coefficient
0.0092(13)



Largest diff. peak and hole
0.152 and −0.162 e. Å−3










Atomic coordinates (×104) of non-H atoms and equivalent isotropic displacement parameters (A2×103) for form N-1. U(eq) is defined as one third of the trace of the orthogonalized Uij tensor.
















x
y
z
U(eq)






















O(1)
−842(2)
7475(1)
5004(1)
67(1)



O(2)
2315(2)
9722(1)
4174(1)
76(1)



O(3)
3875(2)
8859(1)
3393(1)
84(1)



O(4)
4352(2)
7289(1)
2936(1)
89(1)



N(1)
1280(2)
8360(1)
4420(1)
52(1)



N(2)
1830(2)
6969(1)
3995(1)
53(1)



N(3)
3265(3)
5996(1)
3246(1)
72(1)



F(1)
8059(2)
2933(1)
3663(1)
107(1)



C(1)
1120(2)
7423(1)
4351(1)
50(1)



C(2)
104(2)
6877(1)
4711(1)
54(1)



C(3)
−100(3)
8227(1)
5239(1)
68(1)



C(4)
431(3)
8866(1)
4818(1)
64(1)



C(5)
2211(2)
8885(1)
4107(1)
57(1)



C(6)
2989(2)
8365(1)
3710(1)
60(1)



C(7)
2767(2)
7441(1)
3668(1)
56(1)



C(8)
995(3)
6273(1)
5086(1)
67(1)



C(9)
−892(3)
6281(2)
4376(1)
74(1)



C(10)
3526(3)
6895(2)
3256(1)
66(1)



C(11)
3887(3)
5375(2)
2849(1)
86(1)



C(12)
5008(3)
4730(2)
3072(1)
68(1)



C(13)
6169(3)
5043(2)
3373(1)
88(1)



C(14)
7199(3)
4445(2)
3571(1)
90(1)



C(15)
7055(3)
3532(2)
3466(1)
76(1)



C(16)
5953(3)
3190(2)
3169(1)
77(1)



C(17)
4929(3)
3798(2)
2971(1)
73(1)










Example 325










Crystal data for form N-2.
















Temperature
293(2) K


Wavelength
1.54178 Å


Crystal system, space group
Monoclinic, P21/c


Unit cell dimensions
a = 10.2674(2)Å α = 90°



b = 13.2021(2)Å β = 92.0520(10)°



c = 12.5356(2)Å γ = 90°


Volume
1698.13(5)Å3


Z, Calculated density
4, 1.359 Mg/m3


Absorption coefficient
0.889 mm−1


F(000)
728


Crystal size
0.40 × 0.20 × 0.20 mm


Theta range for data collection
4.31 to 63.87°


Limiting indices
−11 <= h <= 11, −15 <= k <= 15,



−13 <= l <= 13


Reflections collected/unique
13897/2697 [R(int) = 0.0344]


Completeness to theta = 25.03
96.0%


Absorption correction
None


Refinement method
Full-matrix least-squares on F2


Data/restraints/parameters
2697/0/229


Goodness-of-fit on F2
1.024


Final R indices [I > 2[](I)]
R1 = 0.0578, wR2 = 0.1481


R indices (all data)
R1 = 0.0991, wR2 = 0.1754


Extinction coefficient
0.0032(5)


Largest diff. peak and hole
0.296 and −0.311 e. Å−3









Atomic coordinates (×104) of non-H atoms and equivalent isotropic displacement parameters (A2×103) for form N-2. U(eq) is defined as one third of the trace of the orthogonalized Uij tensor.
















x
y
z
U(eq)






















O(1)
54(3)
186(3)
1520(2)
156(2)



O(2)
2003(3)
613(2)
5062(2)
99(1)



O(3)
3820(3)
2111(2)
4984(2)
90(1)



O(4)
4669(2)
3577(2)
3876(2)
85(1)



N(1)
1426(2)
861(2)
3301(2)
68(1)



N(2)
2396(2)
2119(2)
2284(2)
66(1)



N(3)
3936(2)
3789(2)
2168(2)
71(1)



F(1)
2445(3)
8332(2)
3172(3)
181(1)



C(1)
1613(3)
1360(2)
2356(2)
65(1)



C(2)
958(3)
966(3)
1326(2)
81(1)



C(3)
−377(5)
12(5)
2401(4)
152(2)



C(4)
367(4)
113(3)
3400(3)
86(1)



C(5)
2166(3)
1079(3)
4227(2)
74(1)



C(6)
3081(3)
1891(3)
4107(2)
69(1)



C(7)
3119(3)
2407(2)
3174(2)
62(1)



C(8)
280(4)
1821(3)
727(3)
104(1)



C(9)
2006(4)
490(3)
654(3)
99(1)



C(10)
3970(3)
3303(2)
3091(2)
67(1)



C(11)
4730(3)
4679(3)
1966(3)
83(1)



C(12)
4101(3)
5657(3)
2282(3)
77(1)



C(13)
4167(3)
5973(3)
3328(3)
93(1)



C(14)
3597(4)
6864(4)
3628(4)
116(2)



C(15)
2988(5)
7426(4)
2883(6)
120(2)



C(16)
2869(5)
7154(4)
1836(5)
131(2)



C(17)
3443(4)
6254(3)
1531(3)
104(1)

## Claims

1. A crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido [2,1-c][1,4]oxazine-2-carboxamide,



characterized by a powder x-ray diffraction pattern comprising 2θ values selected from 7.0±0.1, 11.9±0.1, 17.0±0.1, 18.8±0.1, and 23.9±0.1.

2. The crystalline form of claim 1 which is substantially pure.

3. A crystalline form of claim 1 characterized by a powder X-ray diffraction pattern substantially the same as that of FIG. 1.

4. A crystalline form of claim 1 characterized by a differential scanning calorimetry thermogram substantially the same as that of FIG. 2.

5. A crystalline form of claim 1 characterized by a thermal gravimetric analysis scan substantially the same as that of FIG. 3.

6. A crystalline form of claim 1 characterized by a SSNMR spectra substantially the same as that of FIG. 4.

7. A crystalline form of claim 1 characterized by a moisture sorption isotherm substantially the same as that of FIG. 5.

8. A crystalline form of claim 1 characterized by the unit cell parameters substantially equal to the following criteria: cell dimensions: a=9.2120 Å; b=14.4866 Å; c=25.1791 Å; α=90°; β=90°; γ=90°; space group=Pbca; molecules/unit cell=8; wherein the measurement of the form is at a temperature between about 20° C. to about 25° C.

9. A crystalline form of N-[(4-fluorophenyl)methyl]-4,6,7,9-tetrahydro-3-hydroxy-9,9-dimethyl-4-oxo-pyrimido [2,1-c][1,4]oxazine-2-carboxamide,



characterized by a powder x-ray diffraction pattern comprising 2θ values selected from 8.6±0.1, 10.9±0.1, 15.2±0.1, 15.9±0.1, 19.6±0.1, 21.9±0.1, and 25.8±0.1.

10. A crystalline form of claim 9 which is substantially pure.

11. A crystalline form of claim 1 characterized by a powder X-ray diffraction pattern substantially the same as that of FIG. 6.

12. A crystalline form of claim 9 characterized by a differential scanning calorimetry thermogram substantially the same as that of FIG. 7.

13. A crystalline form of claim 9 characterized by a thermal gravimetric analysis scan substantially the same as that of FIG. 8.

14. A crystalline form of claim 9 characterized by the unit cell parameters substantially equal to the following criteria: cell dimensions: a=10.2674 Å; b=13.2021 Å; c=12.5356 Å; α=90°; β=92.05°; γ=90°; space group=P21/c; molecules/unit cell=4; wherein the measurement of the form is at a temperature between about 20° C. to about 25° C.