# Patent

## Title
Compounds for the treatment of hepatitis C

## Abstract

The disclosure provides compounds of formula I, including pharmaceutically acceptable salts, as well as compositions and methods of using the compounds. The compounds have activity against hepatitis C virus (HCV) and may be useful in treating those infected with HCV.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the benefit of U.S. Provisional Application Ser. No. 61/595,239 filed Feb. 6, 2012.


BACKGROUND OF THE INVENTION
The disclosure generally relates to the novel compounds of formula I including pharmaceutically acceptable salts, which have activity against hepatitis C virus (HCV) and are useful in treating those infected with HCV. The disclosure also relates to compositions and methods of using these compounds.
Hepatitis C virus (HCV) chronically infects an estimated 170 million people worldwide, with 3 to 4 million infected individuals in the United States alone (Boyer, N. and Marcellin, P. J. Hepatology. 2000, 32:98-112; Alter, M. J., et al. Engl. J. Med. 1999, 341:556-562). Prior to the mid 1990s, transfusion with infected blood products was the main route of HCV transmission. Following the introduction of blood screening methods, transmission via injection drug use became the primary risk factor. Chronic infection often leads to the development of severe liver complications, including fibrosis, cirrhosis, and hepatocellular carcinoma. HCV infection is also the leading cause of orthotopic liver transplantation in the United States. The degree to which disease progression is related to viral and cellular factors is not completely understood.
Considerable heterogeneity is found within the nucleotide and encoded amino acid sequence of the HCV genome (Simmonds, P. J. Gen. Virology. 2004, 85:3173-3188). Based on this sequence diversity, six major genotypes and multiple associated subtypes have been described. The genotypes of HCV differ in their worldwide distribution, and the clinical significance of the genetic heterogeneity of HCV remains elusive despite numerous studies of the possible effect of genotypes on pathogenesis and therapy.
Medical treatment for HCV is limited by the lack of a vaccine or approved therapies that specifically target the virus. Currently, patients undergo treatment with a combination of parenterally administered pegylated alpha-interferon and oral ribavirin. Genotype 1 HCV is the most difficult to treat and elimination of the virus (sustained virologic response) is achieved for only approximately 50% of patients (Fried, M. W. et al. N. Engl. J. Med. 2002, 347:975-982; Zeumzem, S, Nature Clinical Practice. 2008, 5:610-622). This poor treatment response, combined with often severe side effects induced by therapy, highlight a need for improved antiviral drugs with better efficacy and safety profiles.
HCV is a member of the Flaviviridae family of viruses with a single-stranded positive-sense RNA genome. Following infection of host cells, the 9.6 Kb genome is translated into a polyprotein precursor of approximately 3,000 amino acids (reviewed in Lindenbach, B. D. and Rice, C. M. Nature. 2005, 436:933-938; Moradpour, D, Penin, F., and Rice, C. M. Nature Reviews. 2007, 5:453-463). Post-translational processing by both cellular and viral proteases results in the generation of at least 10 separate viral proteins. The structural proteins (which by definition are found in mature virions) include core, E1, E2, and possibly p7, and originate from the amino-terminal region of the polyprotein. The core protein assembles into the viral nucleocapsid. The E1 and E2 glycoproteins form heterodimers that are found within the lipid envelope surrounding the viral particles, and mediate host cell receptor binding and entry of the virus into cells. It is unclear if p7 is a structural protein, and its role in replication has yet to be defined. However p7 is believed to form an ion channel in cellular membranes, preventing acidification of intracellular compartments in which virions are assembled, and it has been shown to be essential for viral replication and assembly. The nonstructural proteins NS2, NS3, NS4A, NS4B, NS5A, and NS5B are produced through maturational cleavages of the carboxy-terminal region of the polyprotein. NS2 along with the amino terminus of NS3 form the NS2-3 metalloprotease which cleaves at the NS2-NS3 junction. Additionally, NS2 is involved in assembly and egress of nascent virions. The NS3 protein contains both a serine protease in its amino-terminal region, and a nucleotide-dependent RNA helicase in its carboxy-terminal region. NS3 forms a heterodimer with the NS4A protein, constituting the active protease which mediates cleavages of the polyprotein downstream of NS3, both in cis, at the NS3-NS4A cleavage site, and in trans, for the remaining NS4A-NS4B, NS4B-NS5A, NS5A-NS5B sites. The complex formation of the NS3 protein with NS4A seems necessary to the processing events, enhancing the proteolytic efficiency at all of the sites. The NS3 protein also exhibits nucleoside triphosphatase and RNA helicase activities. The NS4B protein has been shown to be important for localization of HCV proteins into replication complexes in altered membranous structures within the cell. NS5B encodes an RNA-dependent RNA polymerase that is involved in the replication of HCV.
Subgenomic HCV replicons, containing the untranslated regions 5′ and 3′ to the coding sequence fused to the nonstructural proteins or the full-length polyprotein, are competent for translation, viral protein expression, and replication within cultured cells (Lohmann, V. et al. Science. 1999, 285:110-113; Moradpour, D, Penin, F., and Rice, C. M. Nature Reviews. 2007, 5:453-463). The replicon system has proven valuable for the identification of inhibitors targeting the nonstructural proteins associated with these functions. However, only limited subsets of HCV genotypes have been used to generate functional replicons.
Other systems have been used to study the biology of the HCV structural proteins that mediate the entry into host cells. For example, virus-like-particles made in recombinant baculovirus-infected cells with the HCV core, E1 and E2 proteins have also been used to study the function of the HCV E1 and E2 proteins (Barth, H., et al. J. Biol. Chem. 2003, 278:41003-41012). In addition, pseudotyping systems where the E1 and E2 glycoproteins are used to functionally replace the glycoproteins of retroviruses have been developed (Bartosch, B., Dubuisson, J. and Cosset, F.-L. J. Exp. Med. 2003, 197:633-642; Hsu, M. et al. Proc. Natl. Acad. Sci. USA. 2003, 100:7271-7276). These systems yield HCV pseudoparticles that bind to and enter host cells in a manner which is believed to be analogous to the natural virus, thus making them a convenient tool to study the viral entry steps as well as to identify inhibitors block this process.
Recently, a full-length genotype 2a HCV clone, JFH1, was isolated and demonstrated the ability to replicate in vitro. Through repeated passage and adaptation in cell culture increased titers of infectious virus were produced (Lindenbach, B. D., et al. Science. 2005, 309:623-626; Wakita, T. et al. Nature Med. 2005, 11:791-796). In contrast to the HCV replicon or pseudotyping systems, the infectious virus is useful for studying the complete HCV replication cycle, including identifying inhibitors of not only the replication proteins, but those involved in early steps in virus infection (entry and uncoating) and production of progeny viruses (genome packaging, nucleocapsid assembly, virion envelopment and egress).
Triazines have been disclosed. See WO 2009/091388 and US 2009/0286778.
The invention provides technical advantages, for example, the compounds are novel and are effective against hepatitis C. Additionally, the compounds provide advantages for pharmaceutical uses, for example, with regard to one or more of their mechanism of action, binding, inhibition efficacy, target selectivity, solubility, safety profiles, or bioavailability.


DESCRIPTION OF THE INVENTION
One aspect of the invention is a compound of formula I




where
R1 is alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl, alkylcarbonyl, or benzyl wherein the benzyl moiety is substituted with 0-3 substituents selected from halo, alkyl, cycloalkyl, alkenyl, alkynyl, hydroxy, cyano, haloalkyl, alkoxy, and haloalkoxy;
R2 is alkyl, cycloalkyl, (Ar2)alkyl, (Ar2)cycloalkyl, ((Ar2)cycloalkyl)alkyl, ((Ar2)alkyl)cycloalkyl, or (((Ar2)alkyl)cycloalkyl)alkyl;
R3 is hydrogen, alkyl or cycloalkyl;
R4 is hydrogen, alkyl or cycloalkyl;
R5 is




where ring A is a 4 to 7 membered alkylene ring;

R5a is alkyl or benzyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, and alkoxy;
R6 is hydrogen, alkyl or cycloalkyl;
R7 is hydroxy, alkyloxy, phenoxy, SO2R9, SO2N(R10)(R11), CN, alkyl, cycloalkyl, benzocycloalkyl, bicyclicalkyl, (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl, or bridged bicycloalkyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, benzocycloalkyl, bicyclicalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, ether, cyclicether, benzocyclicether, bicyclicether, CO2R9, NR9CO2R11, N(R10)(R11), CON(R10)(R11), NR9CON(R10)(R11), SO2N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, N(R13)COAr3, and N(R13)SO2Ar3;
or R7 is hydrogen, N-alkoxycarbonylpiperidinyl, piperidinonyl, or Ar4;
R8 is hydrogen, alkyl, or cycloalkyl, and alkyl or cycloalkyl is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, fused bicyclic alkyl, bridged bicyclic alkyl, spiro bicyclic alkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, CO2R9, N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, and N(R13)SO2Ar3;
or R8 is substituted with biotin, flurescein or rhosamin;
or R7 and R8 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl;
R9 is hydrogen, Ar3, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, ((hydroxyalkyl)alkoxy)alkoxy, or ((alkoxy)alkoxy)alkoxy;
R10 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, alkoxycarbonyl, or Ar6;
R11 is hydrogen, alkyl, cycloalkyl, or Ar6;
or R10 and R11 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl;
R12 is hydrogen, alkyl, cycloalkyl, or Ar6;
R13 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, alkoxycarbonyl, or Ar6, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, and haloalkoxy, N(R15)(R16), and alkylCO;
R14 is hydrogen, alkyl, cycloalkyl, or Ar6;
or R13 and R14 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl;
R15 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, or alkoxycarbonyl;
R16 is hydrogen, alkyl, or cycloalkyl;
or R15 and R16 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl;
L is alkylene, cycloalkylene, (cycloalkyl)alkyl, (alkyl)cycloalkyl, or alkyl(cycloalkyl)alkyl, and is substituted with 0-2 substituents selected from alkyl, alkoxy, hydroxy, CO2R12 and CONR13R14;
Ar1 is phenyl substituted with 1 CON(R5)(R6) and with 0-3 substituents selected from Ar3, hydroxy, halo, alkyl, cycloalkyl, haloalkyl, alkoxy, and haloalkoxy;
Ar2 is phenyl substituted with 0-3 substituents selected from halo, hydroxy, cyano, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, alkoxy, and haloalkoxy;
Ar3 is phenyl, biphenyl, terphenyl, naphthalenyl, furanyl, benzofuranyl, fluorenyl, fluorenonyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzoisothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, indolinyl, chromenonyl, or dibenzofuranyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, Ar5, OAr5, NR13Ar5, N(R13)COAr5, N(R13)SO2Ar5, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), N(R13)(R14), amidine, urea, ketone, sulfone, sulfamide, and PhCONHSO2; and said alkyl, alkenyl, cycloalkyl, alkynyl or Ar5 is further substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, aryoxy, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), N(R13)(R14), amidine, urea, ketone, sulfone, sulfamide, PhCONHSO2 and Ar6;
or Ar3 is phenyl substituted with 1 substituents selected from benzyl, phenoxy, pyridyloxy, pyrimidyloxy, tetrazolyloxy, thiazolyl, phenylpyrazolyl, methyloxadiazolyl, thiadiazolyl, triazolyl, methyltriazolyl, tetrazolyl, pyridinyl, dimethoxypyrimdinyl, indolyl, indolinyl, and isoindolinyl;
Ar4 is phenyl, indanyl, tetrahydronaphthyl, isochromanyl, benzodioxolyl, pyridinyl, pyrazolyl, imidazolyl, or triazolyl, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, and haloalkoxy, N(R13)(R14), and alkylCO;
Ar5 is phenyl, naphthalenyl, furanyl, benzofuranyl, azabenzofuranyl, thiophenyl, benzothiophenyl, azabenzothiophenyl, pyrrolyl, indolyl, azaindolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindazolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, or indolinyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, OAr6, NR13Ar6, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), N(R13)(R14), amidine, urea, ketone, sulfone and sulfamide;
or Ar5 is substituted with biotin, flurescein or rhosamin;
Ar6 is phenyl, naphthalenyl, furanyl, benzofuranyl, azabenzofuranyl, thiophenyl, benzothiophenyl, azabenzothiophenyl, pyrrolyl, indolyl, azaindolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindazolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, or indolinyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, phenyl, hydroxy, alkoxy, aryloxy, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, ester, ketone, amidine, urea, ketone, sulfone and sulfamide;
or Ar6 is substituted with biotin, flurescein or rhosamin;
or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where:
R1 is alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl, alkylcarbonyl, or benzyl wherein the benzyl moiety is substituted with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy, and haloalkoxy;
R2 is alkyl, (Ar2)alkyl, (Ar2)cycloalkyl, ((Ar2)cycloalkyl)alkyl, ((Ar2)alkyl)cycloalkyl, or (((Ar2)alkyl)cycloalkyl)alkyl;
R3 is hydrogen or alkyl;
R4 is hydrogen or alkyl;
R5 is




where ring A is a 4 to 7 membered alkylene ring substituted with L;

R6 is hydrogen or alkyl;
R7 is alkyl, cycloalkyl, (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl, or a bridged bicycloalkyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, CO2R9, N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, and Ar4;
or R7 is hydrogen, N-alkoxycarbonylpiperidinyl, piperidinonyl, or Ar3;
R8 is hydrogen or alkyl;
or R7 and R8 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl;
R9 is hydrogen, alkyl, hydroxyalkyl, alkoxyalkyl, ((hydroxyalkyl)alkoxy)alkoxy, or ((alkoxy)alkoxy)alkoxy;
R10 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, or alkoxycarbonyl;
R11 is hydrogen, alkyl;
or R10 and R11 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl;
R12 is hydrogen or alkyl;
R13 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, or alkoxycarbonyl;
R14 is hydrogen or alkyl;
or R13 and R14 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl;
L is alkylene, cycloalkylene, (cycloalkyl)alkyl, (alkyl)cycloalkyl, or alkyl(cycloalkyl)alkyl, and is substituted with 0-1 CO2R12 or CONR13R14;
Ar1 is phenyl substituted with 1 CON(R5)(R6) and with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy, and haloalkoxy;
Ar2 is phenyl substituted with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy, and haloalkoxy;
Ar3 is phenyl, indanyl, fluorenyl, biphenyl, terphenyl, pyridinyl, pyrazolyl, isoxazolyl, imidazolyl, thiazolyl, triazolyl, benzoxazolyl, indolinyl, or dibenzofuranyl, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkenyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14)))alkyl, phenyl, hydroxy, alkoxy, haloalkoxy, alkylcarbonyl, CO2R12, CON(R13)(R14), or PhCONHSO2;
or Ar3 is phenyl substituted with 1 substituents selected from benzyl, tetrazolyloxy, thiazolyl, phenylpyrazolyl, methyloxadiazolyl, thiadiazolyl, triazolyl, methyltriazolyl, tetrazolyl, pyridinyl, dimethoxypyrimdinyl; and
Ar4 is phenyl, indanyl, tetrahydronaphthyl, isochromanyl, benzodioxolyl, pyridinyl, pyrazolyl, or imidazolyl, triazolyl and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, and haloalkoxy, N(R13)(R14), and alkylCO;
or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R1 is alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl, alkylcarbonyl, or benzyl.
Another aspect of the invention is a compound of formula I where R1 is alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, benzyl, indanyl, or alkylcarbonyl.
Another aspect of the invention is a compound of formula I where R2 is alkyl, cycloalkyl, (Ar2)alkyl, (Ar2)cycloalkyl, ((Ar2)cycloalkyl)alkyl, ((Ar2)alkyl)cycloalkyl, or (((Ar2)alkyl)cycloalkyl)alkyl.
Another aspect of the invention is a compound of formula I where R2 is alkyl, (Ar2)alkyl, (Ar2)cycloalkyl, ((Ar2)cycloalkyl)alkyl, ((Ar2)alkyl)cycloalkyl, or (((Ar2)alkyl)cycloalkyl)alkyl.
Another aspect of the invention is a compound of formula I where R3 is hydrogen, alkyl or cycloalkyl.
Another aspect of the invention is a compound of formula I where R3 is hydrogen or alkyl.
Another aspect of the invention is a compound of formula I where R3 is hydrogen.
Another aspect of the invention is a compound of formula I where R4 is hydrogen, alkyl or cycloalkyl.
Another aspect of the invention is a compound of formula I where R4 is hydrogen or alkyl.
Another aspect of the invention is a compound of formula I where R4 is hydrogen.
Another aspect of the invention is a compound of formula I where R5 is




where ring A is a 4 to 7 membered alkylene ring substituted with L.

Another aspect of the invention is a compound of formula I where R5 is




where ring A is a 4 to 7 membered alkylene ring substituted with L.

Another aspect of the invention is a compound of formula I where R5a is alkyl or benzyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, and alkoxy.
Another aspect of the invention is a compound of formula I where R6 is hydrogen, alkyl or cycloalkyl.
Another aspect of the invention is a compound of formula I where R6 is hydrogen or alkyl.
Another aspect of the invention is a compound of formula I where R7 is hydroxy, alkyloxy, phenoxy, SO2R9, SO2N(R10)(R11), CN, alkyl, cycloalkyl, benzocycloalkyl, bicyclicalkyl, (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl, or a bridged bicycloalkyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, benzocycloalkyl, bicyclicalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, ether, cyclicether, benzocyclicether, bicyclicether, CO2R9, NR9CO2R11, N(R10)(R11), CON(R10)(R11), NR9CON(R10)(R11), SO2N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, N(R13)COAr3, and N(R13)SO2Ar3;
Another aspect of the invention is a compound of formula I where R7 is alkyl, cycloalkyl, (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl, or a bridged bicycloalkyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, CO2R9, N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, and Ar4.
Another aspect of the invention is a compound of formula I where R7 is hydrogen, N-alkoxycarbonylpiperidinyl, piperidinonyl, or Ar4.
Another aspect of the invention is a compound of formula I where R7 is hydrogen, N-alkoxycarbonylpiperidinyl, piperidinonyl, or Ar3.
Another aspect of the invention is a compound of formula I where R8 is hydrogen, alkyl or cycloalkyl, and said alkyl or cycloalkyl is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, fused bicyclic alkyl, bridged bicyclic alkyl, spiro bicyclic alkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, CO2R9, N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, and N(R13)SO2Ar3.
Another aspect of the invention is a compound of formula I where R8 is hydrogen or alkyl.
Another aspect of the invention is a compound of formula I where R8 is substituted with biotin, flurescein or rhosamin.
Another aspect of the invention is a compound of formula I where R7 and R8 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R7 and R8 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R9 is hydrogen, Ar3, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, ((hydroxyalkyl)alkoxy)alkoxy, or ((alkoxy)alkoxy)alkoxy.
Another aspect of the invention is a compound of formula I where R9 is hydrogen, alkyl, hydroxyalkyl, alkoxyalkyl, ((hydroxyalkyl)alkoxy)alkoxy, or ((alkoxy)alkoxy)alkoxy.
Another aspect of the invention is a compound of formula I where R10 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, alkoxycarbonyl, or Ar6.
Another aspect of the invention is a compound of formula I where R10 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, or alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R11 is hydrogen, alkyl, cycloalkyl, or Ar6.
Another aspect of the invention is a compound of formula I where R11 is hydrogen or alkyl.
Another aspect of the invention is a compound of formula I where R10 and R11 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R10 and R11 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R12 is hydrogen, alkyl, cycloalkyl, or Ar6.
Another aspect of the invention is a compound of formula I where R12 is hydrogen or alkyl.
Another aspect of the invention is a compound of formula I where R13 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, alkoxycarbonyl, or Ar6 and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, haloalkoxy, N(R15)(R16), and alkylCO.
Another aspect of the invention is a compound of formula I where R13 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, or alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R14 is hydrogen, alkyl, cycloalkyl, or Ar6.
Another aspect of the invention is a compound of formula I where R14 is hydrogen or alkyl;
Another aspect of the invention is a compound of formula I where R13 and R14 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R13 and R14 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R15 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, or alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where R16 is hydrogen, alkyl or cycloalkyl.
Another aspect of the invention is a compound of formula I where R15 and R16 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, (aryl)alkyl, alkylcarbonyl, and alkoxycarbonyl.
Another aspect of the invention is a compound of formula I where L is alkylene, cycloalkylene, (cycloalkyl)alkyl, (alkyl)cycloalkyl, or alkyl(cycloalkyl)alkyl, and is substituted with 0-2 substituents selected from alkyl, alkoxy, hydroxy, CO2R12 or CONR13R14.
Another aspect of the invention is a compound of formula I where L is alkylene, cycloalkylene, (cycloalkyl)alkyl, (alkyl)cycloalkyl, or alkyl(cycloalkyl)alkyl, and is substituted with 0-1 CO2R12 or CONR13R14.
Another aspect of the invention is a compound of formula I where Ar1 is phenyl substituted with 1 CON(R5)(R6) and with 0-3 substituents selected from Ar3, hydroxy, halo, alkyl, cycloalkyl, haloalkyl, alkoxy, and haloalkoxy.
Another aspect of the invention is a compound of formula I where Ar2 is phenyl substituted with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy, and haloalkoxy.
Another aspect of the invention is a compound of formula I where Ar1 is fluorophenyl substituted with 1 CON(R5)(R6) and with 0-3 substituents selected from Ar3, hydroxy, halo, alkyl, cycloalkyl, haloalkyl, alkoxy, and haloalkoxy.
Another aspect of the invention is a compound of formula I where Ar2 is phenyl substituted with 0-3 substituents selected from halo, hydroxy, cyano, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, alkoxy, and haloalkoxy.
Another aspect of the invention is a compound of formula I where Ar2 is phenyl substituted with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy, and haloalkoxy.
Another aspect of the invention is a compound of formula I where Ar3 is phenyl, biphenyl, terphenyl, naphthalenyl, furanyl, benzofuranyl, fluorenyl, fluorenonyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzoisothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, indolinyl, chromenonyl, or dibenzofuranyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, Ar5, OAr5, NR13, Ar5, N(R13)COAr5, N(R13)SO2Ar5, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), N(R13)(R14), amidine, urea, ketone, sulfone, sulfamide, and PhCONHSO2; and said alkyl, alkenyl, cycloalkyl, alkynyl or Ar5 is further substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, aryoxy, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), N(R13)(R14), amidine, urea, ketone, sulfone, sulfamide, PhCONHSO2 and Ar6.
Another aspect of the invention is a compound of formula I where Ar3 is phenyl, indanyl, fluorenyl, biphenyl, terphenyl, pyridinyl, pyrazolyl, isoxazolyl, imidazolyl, thiazolyl, triazolyl, benzoxazolyl, indolinyl, or dibenzofuranyl, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkenyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, haloalkoxy, alkylcarbonyl, CO2R12, CON(R13)(R14), or PhCONHSO2.
Another aspect of the invention is a compound of formula I where Ar3 is phenyl substituted with 1 substituents selected from benzyl, phenoxy, pyridyloxy, pyrimidyloxy, tetrazolyloxy, thiazolyl, phenylpyrazolyl, methyloxadiazolyl, thiadiazolyl, triazolyl, methyltriazolyl, tetrazolyl, pyridinyl, dimethoxypyrimdinyl, indolyl, indolinyl, and isoindolinyl.
Another aspect of the invention is a compound of formula I where Ar3 is phenyl substituted with 1 substituents selected from benzyl, tetrazolyloxy, thiazolyl, phenylpyrazolyl, methyloxadiazolyl, thiadiazolyl, triazolyl, methyltriazolyl, tetrazolyl, pyridinyl, and dimethoxypyrimdinyl.
Another aspect of the invention is a compound of formula I where Ar4 is phenyl, indanyl, tetrahydronaphthyl, isochromanyl, benzodioxolyl, pyridinyl, pyrazolyl, imidazolyl, or triazolyl, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, and haloalkoxy, N(R13)(R14), and alkylCO.
Another aspect of the invention is a compound of formula I where Ar5 is phenyl, naphthalenyl, furanyl, benzofuranyl, azabenzofuranyl, thiophenyl, benzothiophenyl, azabenzothiophenyl, pyrrolyl, indolyl, azaindolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindazolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, or indolinyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, OAr6, NR13Ar6, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), N(R13)(R14), amidine, urea, ketone, sulfone and sulfamide.
Another aspect of the invention is a compound of formula I where Ar5 is substituted with biotin, flurescein, or rhosamin.
Another aspect of the invention is a compound of formula I where Ar6 is phenyl, naphthalenyl, furanyl, benzofuranyl, azabenzofuranyl, thiophenyl, benzothiophenyl, azabenzothiophenyl, pyrrolyl, indolyl, azaindolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindazolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, or indolinyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, phenyl, hydroxy, alkoxy, aryloxy, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, ester, ketone, amidine, urea, ketone, sulfone and sulfamide
Another aspect of the invention is a compound of formula I where Ar6 is substituted with biotin, flurescein, or rhosamin;
Another aspect of the invention is a compound of formula I where R1 is haloalkyl or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R1 is trifluoroethyl or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R1 is alkyl or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R1 is phenylalkyl substituted with 0-5 halo, alkyl, alkenyl, haloalkyl, alkoxy or haloalkoxy or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R1 is benzyl substituted with 0-5 halo, alkyl, alkenyl, haloalkyl, alkoxy or haloalkoxy or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R2 is (Ar2)alkyl or (Ar2)cycloalkyl, or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R2 is (Ar2)alkyl or (Ar2)cycloalkyl, or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where R7 is hydroxy, alkyloxy, phenoxy, SO2R9, SO2N(R10)(R11), CN, alkyl, cycloalkyl, benzocycloalkyl, bicyclicalkyl, (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl, or a bridged bicycloalkyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, benzocycloalkyl, bicyclicalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, ether, cyclicether, benzocyclicether, bicyclicether, CO2R9, NR9CO2R11, N(R10)(R11), CON(R10)(R11), NR9CON(R10)(R11), SO2N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, N(R13)COAr3, and N(R13)SO2Ar3.
Another aspect of the invention is a compound of formula I where R7 is Ar4 or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where L is




or a pharmaceutically acceptable salt thereof.

Another aspect of the invention is a compound of formula I where Ar1 is phenyl para-substituted with 1 CON(R5)(R6), or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a compound of formula I where Ar1 is fluoro phenyl para-substituted with 1 CON(R5)(R6), or a pharmaceutically acceptable salt thereof.
Any scope of any variable, including R1, R2, R3, R4, R5, R5a, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15, R16, L, Ar1, Ar2, Ar3, Ar4, Ar5, and Ar6 can be used independently with the scope of any other instance of a variable.
Unless specified otherwise, these terms have the following meanings. “Halo” means fluoro, chloro, bromo, or iodo. “Alkyl” means a straight or branched alkyl group composed of 1 to 6 carbons. “Alkenyl” means a straight or branched alkyl group composed of 2 to 6 carbons with at least one double bond. “Alkynyl” means a straight or branched alkyl group composed of 2 to 6 carbons with at least one triple bond. “Cycloalkyl” means a monocyclic ring system composed of 3 to 8 carbons. “Benzocycloalkyl” means a monocyclic ring system composed of 3 to 8 carbons fused to a benzene ring system. “Bicycloalkyl” means a fused bicyclic ring system wherein each ring is composed of 3 to 7 carbons, that is a [(3-7).0.(3-7)] system. “Bridged bicycloalkyl” means a [(3-7).(1-3).(3-7)] ring system. “Spirocycloalkyl” means a spirocyclic ring system wherein each ring is composed of 3 to 7 carbons. “Alkylene” means a straight or branched divalent alkyl group. “Alkenylene” means a straight or branched divalent alkyl group with at least one double bond. “Cycloalkylene” means a divalent cycloalkane moiety composed of 3 to 7 carbons and includes gem-divalency (for example 1,1-cyclopropanediyl) as well as non-gem-divalency (for example, 1,4-cyclohexanediyl). “Alkylidinyl” means a divalent alkene substituent where the divalency occurs on the same carbon of the alkene. “Hydroxyalkyl,” “alkoxy” and other terms with a substituted alkyl moiety include straight and branched isomers composed of 1 to 6 carbon atoms for the alkyl moiety. “Haloalkyl” and “haloalkoxy” include all halogenated isomers from monohalo substituted alkyl to perhalo substituted alkyl. “Aryl” includes carbocyclic and heterocyclic aromatic substituents. Phenylene is a divalent benzene ring. “1,4-Phenylene” means 1,4-benzenediyl with respect to regiochemistry for the divalent moiety. Parenthetic and multiparenthetic terms are intended to clarify bonding relationships to those skilled in the art. For example, a term such as ((R)alkyl) means an alkyl substituent further substituted with the substituent R.
The substituents described above may be attached at any suitable point of attachment unless otherwise specified. However, it is understood that the compounds encompassed by the present invention are those that are chemically stable as understood by those skilled in the art. Additionally, the compounds encompassed by the present disclosure are those that are suitably stable for use as a pharmaceutical agent.
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, camsylate, 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.
Some of the compounds of the invention possess asymmetric carbon atoms (see, for example, the structures below). The invention includes all stereoisomeric forms, including enantiomers and diastereomers as well as mixtures of stereoisomers such as racemates. Some stereoisomers can be made using methods known in the art. Stereoisomeric mixtures of the compounds and related intermediates can be separated into individual isomers according to methods commonly known in the art. The use of wedges or hashes in the depictions of molecular structures in the following schemes and tables is intended only to indicate relative stereochemistry, and should not be interpreted as implying absolute stereochemical assignments.
The invention is intended to include all isotopes of atoms occurring in the present compounds. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include deuterium and tritium. Isotopes of carbon include 13C and 14C. Isotopically-labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed. Such compounds may have a variety of potential uses, for example as standards and reagents in determining biological activity. In the case of stable isotopes, such compounds may have the potential to favorably modify biological, pharmacological, or pharmacokinetic properties.
Biological Methods
Infection Assays.
HCV pseudoparticles, produced using standardized methodology (Bartosch, B., Dubuisson, J. and Cosset, F.-L. J. Exp. Med. 2003, 197:633-642) were made via a liposome-based transfection procedure of 293T cells with plasmids expressing the murine leukemia virus capsid and polymerase proteins, an MLV genome encoding the luciferase reporter gene, and envelope glycoproteins from either HCV or vesicular stomatitis virus (VSV). The genotype 1a HCV E1 and E2 envelope coding sequences were derived from the H77C isolate (GenBank accession number AF009606). Media containing pseudoparticles was collected 3 days following transfection, filtered, and stored at −20° C. as a viral stock. Infections were performed in 384-well plates by mixing pseudovirus with 1×104 Huh7 cells/well in the presence or absence of test inhibitors, followed by incubation at 37° C. Luciferase activity, reflecting the degree of entry of the pseudoparticles into host cells, was measured 2 days after infection. The specificity of the compounds for inhibiting HCV was determined by evaluating inhibition of VSV pseudoparticle infection.
Compounds and Data Analysis.
Test compounds were serially diluted 3-fold in dimethyl sulfoxide (DMSO) to give a final concentration range in the assay of 50.0 μM to 0.04 μM. Maximum activity (100% of control) and background were derived from control wells containing DMSO but no inhibitor or from uninfected wells, respectively. The individual signals in each of the compound test wells were then divided by the averaged control values after background subtraction and multiplied by 100% to determine percent activity. Assays were performed in duplicate and average EC50 values (reflecting the concentration at which 50% inhibition of virus replication was achieved) were calculated. Compound EC50 data is expressed as A=0.01≦10 nM; B=10-1000 nM. Representative data for compounds are reported in Table 1.










TABLE 1





Compound
Structure
EC50
Activity


















1001





B


 


1002




2.11
A


 


1003





A


 


1004




4.74
A


 


1005





A


 


1006




0.69
A


 


1007





A


 


1008





A


 


1009





A


 


1010





A


 


1011





A


 


1012





A


 


1013





A


 


1014





A


 


1015





A


 


1016





A


 


1017





A


 


1018





A


 


1019





A


 


1020





A


 


1021





A


 


1022




7.11
A


 


1023





A


 


1024





A


 


1025




1.73
A


 


1026




2.68
A


 


1027





A


 


1028




0.40
A


 


1029





A


 


1030





A


 


1031





A


 


1032





A


 


1033





A


 


1034





A


 


1035




0.03
A


 


1036





A


 


1037





A


 


1038





A


 


1039





A


 


1040





B


 


1041





A


 


1042





A


 


1043





A


 


1044





A


 


1045





A


 


1046





A


 


1047





A


 


1048





A


 


1049





A


 


1050





A


 


1051





A


 


1051





A


 


1053





A


 


1054





A


 


1055




2.41
A


 


1056





A


 


1057





A


 


1058




1.15
A


 


1059





A


 


1060





A


 


1061




1.58
A


 


1062





A


 


1063





A


 


1064




1.89
A


 


1065




0.59
A


 


1066





A


 


1067





A


 


1068





A


 


1069





A


 


1070





A


 


1071





A


 


1072





A


 


1073





A


 


1074





A


 


1075





A


 


1076





A


 


1077





A


 


1078




11.91
B


 


1079





A


 


1080





A


 


1081





A


 


1082





A


 


1083




1.78
A


 


1084




0.26
A


 


1085





A


 


1086





A


 


1087





A


 


1088





A


 


1089





A


 


1090





A


 


1091





A


 


1092





A


 


1093





A


 


1094





A


 


1095





A


 


1096





A


 


1097





A


 


1098





A


 


1099





A


 


1100





A


 


1101





A


 


1102





A


 


1103





A


 


1104





A


 


1105





A


 


1106





A


 


1107




0.46
A


 


1108





A


 


1109





A


 


1110





A


 


1111





A


 


1112




1.23
A


 


1113





A


 


1114





A


 


1115





A


 


1116





B


 


1117





A


 


1118





A


 


1119





A


 


1120





A


 


1121





A


 


1122





A


 


1123





A


 


1124





A


 


1125





A


 


1126





A


 


1127





A


 


1128





A


 


1129





A


 


1130




0.12
A


 


1131





A


 


1132





A


 


1133




1.40
A


 


1134





A


 


1135




1.92
A


 


1136





A


 


1137





A


 


1138




0.81
A


 


1139





A


 


1140





A


 


1141





A


 


1142





A


 


1143





A


 


1144





A


 


1145





A


 


1146





A


 


1147





A


 


1148





A


 


1149





A


 


1150





A


 


1151




1.62
A


 


1152





A


 


1153





A


 


1154




1.07
A


 


1155





A


 


1156





A


 


1157





A


 


1158





A


 


1159





A


 


1160





A


 


1161





A


 


1162





A


 


1163





A


 


1164





A


 


1165





A


 


1166





A


 


1167




1.32
A


 


1168





A


 


1169





A


 


1170





A


 


1171





A


 


1172




9.26
A


 


1173




0.74
A


 


1174





A


 


1175





A


 


1176





A


 


1177





A


 


1178





A


 


1179





A


 


1180





A


 


1181





A


 


1182





A


 


1183





A


 


1184





A


 


1185





A


 


1186





A


 


1187




0.08
A


 


1188





A


 


1189





A


 


1190





A


 


1191




0.17
A


 


1192





A


 


1193




0.52
A


 


1194





A


 


1195





A


 


1196





A


 


1197





A


 


1198





A


 


1199




0.19
A


 


1200





A


 


1201





A


 


1202





A


 


1203





A


 


1204





A


 


1205





A


 


1206





A


 


1207





A


 


1208




0.21
A


 


1209





A


 


1210





A


 


1211




0.13
A


 


1212




0.07
A


 


1213





A


 


1214





A


 


1215





A


 


1216





A


 


1217





A


 


1218





A


 


1219





B


 


1220





A


 


1221





A


 


1222





A


 


1223





A


 


1224





A


 


1225





A


 


1226





A


 


1227





A


 


1228





A


 


1229





A


 


1230





A


 


1231





A


 


1232





A


 


1233





A


 


1234





A


 


1235




0.53
A


 


1236





A


 


1237




0.77
A


 


1238





A


 


1239





A


 


1240





A


 


1241




0.99
A


 


1242




0.32
A


 


1243





B


 


1244





A


 


1245




1.69
A


 


1246





A


 


1247





A


 


1248





A


 


1249





A


 


1250





A


 


1251





A


 


1252





A


 


1253





A


 


1254




0.37
A


 


1255




1.10
A


 


1256




0.64
A


 


1257





A


 


1258





A


 


1259





A


 


1260





A


 


1261





A


 


1262





A


 


1263





A


 


1264





B


 


1265





A


 


1266





A


 


1267




5.26
A


 


1268





A


 


1269





A


 


1270




0.13
A


 


1271





A


 


1272





A


 


1273





A


 


1274





A


 


1275





A


 


1276





A


 


1277





A


 


1278





A


 


1279




1.29
A


 


1280





A


 


1281





A


 


1282





A


 


1283





A


 


1284





A


 


1285





A


 


1286





A


 


1287





A


 


1288





A


 


1289





A


 


1290





A


 


1291





A


 


1292





A


 


1293




4.40
A


 


1294





A


 


1295





B


 


1296





A


 


1297





A


 


1298





A


 


1299





A


 


1300





A


 


1301





A


 


1302





A


 


1303





A


 


1304





A


 


1305





A


 


1306





A


 


1307




0.38
A


 


1308





A


 


1309





A


 


1310




0.62
A


 


1311





A


 


1312





A


 


1313





A


 


1314




0.43
A


 


1315




1.03
A


 


1316




0.57
A


 


1317





A


 


1318





A


 


1319





A


 


1320





A


 


1321





A


 


1322





A


 


1323




6.28
A


 


1324





A


 


1325





A


 


1326





A


 


1327





A


 


1328





A


 


1329





A


 


1330





A


 


1331





A


 


1332





A


 


1333





A


 


1334





A


 


1335





A


 


1336




0.88
A


 


1337





A


 


1338





A


 


1339





A


 


1340




0.10
A


 


1341





A


 


1342





A


 


1343





A


 


1344





A


 


1345





A


 


1346





A


 


1347





A


 


1348




0.50
A


 


1349





A


 


1350





B


 


1351





A


 


1352





A


 


1353




1.66
A


 


1354





A


 


1356





A


 


1357





A


 


1358





A


 


1359





A


 


1360





A


 


1361





A


 


1362




0.30
A


 


1363





A


 


1364





A


 


1365





A


 


1367





A


 


1368





A


 


1369





A


 


1370





A


 


1371





A


 


1372





A


 


1373





A


 


1374





A


 


1375





A


 


1376





A


 


1377





A


 


1378




0.23
A


 


1379





A


 


1380





A


 


1381





A


 


1382





A


 


1383





A


 


1384





A


 


1385





A


 


1386




2.76
A


 


1387





A


 


1388





A


 


1389





A


 


1390





A


 


1391





A


 


1392




0.14
A


 


1393





A


 


1394





A


 


1395





A


 


1396





A


 


1397





A


 


1398




2.19
A


 


1399





A


 


1400




3.77
A


 


1401





A


 


1402





A


 


1403





A


 


1404





A


 


1405





A


 


1406





A


 


1407





A


 


1408





A


 


1409








 


1410





A


 


1411





A


 


1412





B


 


1413





A


 


1414





A


 


1415





A


 


1416





A


 


1417




0.16
A


 


1418




0.28
A


 


1419





A


 


1420





A


 


1421





A


 


1422





A


 


1423





A


 


1424





A


 


1425





A


 


1426





A


 


1427





A


 


1428





A


 


1429





A


 


1430





A


 


1431





A


 


1432





A


 


1433





A


 


1434





A


 


1435




0.11
A


 


1436





A


 


1437





A


 


1438





A


 


1439





A


 


1440





A


 


1441





A


 


1442





A


 


1443





A


 


1444





A


 


1445





A


 


1446





A


 


1447




97.74
B


 


1448





A


 


1449





A


 


1450





A


 


1451





A


 


1452




1.83
A


 


1453





A


 


1454





A


 


1455





A


 


1456




3.45
A


 


1457





A


 


1458




3.20
A


 


1459





A


 


1460





A


 


1461





A


 


1462





A


 


1464




0.48
A


 


1465





A


 


1466





A


 


1467





A


 


1468





A


 


1469




2.30
A


 


1470





A


 


1471





A


 


1472





A


 


1473





A


 


1474





A


 


1475





A


 


1476





A


 


1477





A


 


1478





A


 


1479





A


 


1480





A


 


1481





A


 


1482




0.17
A


 


1483





A


 


1484





A


 


1485





A


 


1487




0.22
A


 


1488





A


 


1489





A


 


1490





A


 


1491





A


 


1492





A


 


1493





A


 


1494





A


 


1495





A


 


1496





A


 


1497





A


 


1498




0.24
A


 


1499





A


 


1500





A


 


1501





A


 


1502





A


 


1503





A


 


1504




<2.54
A


 


1505





A


 


1506





A


 


1507





A


 


1508





A


 


1509





A


 


1510





A


 


1511





A


 


1512





A


 


1513





A


 


1514




0.04
A


 


1515





A


 


1516




0.09
A


 


1517





A


 


1518





A


 


1519





A


 


1520





A


 


1521





A


 


1522




0.10
A


 


1523




0.26
A


 


1524





A


 


1525





A


 


1526





A


 


1527




0.15
A


 


1528





A


 


1529





A


 


1530





A


 


1532





A


 


1533





A


 


1534





A


 


1535





A


 


1536





A


 


1537





A


 


1538





A


 


1539





A


 


1540





A


 


1541





A


 


1542





A


 


1543





A


 


1544




0.08
A


 


1545





A


 


1546





A


 


1547




0.06
A


 


1548





A


 


1549





A


 


1550





A


 


1551





A


 


1552





A


 


1553





A


 


1554





A


 


1555





A


 


1556





A


 


1557





A


 


1558





A


 


1559





A


 


1560





A


 


1561





A


 


1562





A


 


1563





A


 


1564





A


 


1565




0.13
A


 


1566





A


 


1567





A


 


1568





A


 


1569





A


 


1570




0.05
A


 


1571




0.05
A


 


1572





A


 


1573





A


 


1574





A


 


1576





A


 


1577





A


 


1578





A


 


1579





A


 


1580




0.04
A


 


1581





A


 


1582





A


 


1583





A


 


1584





A


 


1585




0.03
A


 


1586





A


 


1587





A


 


1588





A


 


1589





A


 


1590





A


 


1591





A


 


1592





A


 


1593





A


 


1594




0.05
A


 


1595





A


 


1596





A


 


1597





A


 


1598





A


 


1599





A


 


1600





A


 


1601




0.03
A


 


1602





A


 


1603




0.04
A


 


1604




0.02
A


 


2001





A


 


2002





A


 


3001





A


 


3002




0.044
A


 


3003





A


 


3004





A


 


3005





A


 


3006





A


 


3007





A


 


3008





A


 


3009





A


 


3010




0.014
A


 


3011





A


 


3012





A


 


3013




0.066
A


 


3014





A


 


3015





A


 


3016





A


 


3017





A


 


3018





A


 


3019





A


 


3020





A


 


3021




0.73
A


 


3022





A


 


3023





A


 


3024





A


 


3025





A


 


3026





A


 


3027





A


 


3028





A


 


3029




0.071
A


 


3030





A


 


3031





A


 


3032





A


 


3033





A


 


3034




0.040
A


 


3035





A


 


3036





A


 


3037





A


 


3038





A


 


3039





A


 


3040





A


 


3041




0.068
A


 


3042





A


 


3043





A


 


3044





A


 


3045





A


 


3046





A


 


3047





A


 


3048




0.27
A


 


3049





A


 


3050





A


 


3051





A


 


3052





A


 


3053





A


 


3054





A


 


3055




0.0095
A


 


3056




0.0070
A


 


3057





A


 


3058





A


 


3059





A


 


3060





A


 


3061





A


 


3062





A


 


3063





A


 


3064





A


 


3065





A


 


3066




0.032
A


 


3067





A


 


3068





A


 


3069





A


 


3070





A


 


3071





A


 


3072





A


 


3073





A


 


3074




0.054
A


 


3075





A


 


3076





A


 


3077





A


 


3078





A


 


3079




1.42
A


 


3080





A


 


3081





A


 


3082





A


 


3083





A


 


3084




3.58
A


 


3085





A


 


3086





A


 


3087





A


 


3088





A


 


3089





A


 


3090





A


 


3091




2.85
A


 


3092





A


 


3093





A


 


3094





A


 


3095





A


 


3096





A


 


3097





A


 


3098





A


 


3099





A


 


3100




0.27
A


 


3101





A


 


3102





A


 


3103





A


 


3104





A


 


3105





A


 


3106





A


 


3107




0.92
A


 


3108





A


 


3109





A


 


3110





A


 


3111





A


 


3112





A


 


3113




7.77
A


 


3114




0.084
A


 


3115





A


 


3116





A


 


3117





A


 


3118





A


 


3119





A


 


3120





A


 


3121




3.39
A


 


3122





A


 


3123





A


 


3124




18.18
A


 


3125





A


 


3126





A


 


3127





A


 


3128





A


 


3129




0.089
A


 


3130





A


 


3131





A


 


3132





A


 


3133





A


 


3134





A


 


3135




3.69
A


 


3136





A


 


3137





A


 


3138




0.46
A


 


3139





A


 


3140





A


 


3141




0.11
A


 


3142





A


 


3143





A


 


3144





A


 


3145





A


 


3146




0.050
A


 


3147





A


 


3148





A


 


3149





A


 


3150





A


 


3151





A


 


3152





A


 


3153





A


 


3154




1.44
A


 


3155





A


 


3156





A


 


3157





A


 


3158





A


 


3159





A


 


3160





A


 


3161





A


 


3162





A


 


3163





A


 


3164




0.68
A


 


3165





A


 


3166





A


 


3167





A


 


3168





A


 


3169





A


 


3170




0.63
A


 


3171





A


 


3172





A


 


3173





A


 


3174





A


 


3175




0.11
A


 


3176





A


 


3177





A


 


3178





A


 


3179





A


 


3180





A


 


3181





A


 


3182





A


 


3183





A


 


3184





A


 


3185





A


 


3186




0.70
A


 


3187





A


 


3188





A


 


3189





A


 


3190





A


 


3191




0.82
A


 


3192





A


 


3193




1.03
A


 


3194





A


 


3195





A


 


3196





A


 


3197





A


 


3198




1.02
A


 


3199





A


 


3200





A


 


3201





A


 


3202





A


 


3203




5.47
A


 


3204





A


 


3205





A


 


3206





A


 


3207





A


 


3208





A


 


3209





A


 


3210




0.75
A


 


3211





A


 


3212





A


 


3213





A


 


3214





A


 


3215





A


 


3216





A


 


3217




0.087
A


 


3218





A


 


3219





A


 


3220





A


 


3221





A


 


3222





A


 


3223





A


 


3224





A


 


3225




16.99
B


 


3226





A


 


3227





A


 


3228





A


 


3229





B


 


3230





A


 


3231





A


 


3232





A


 


3233





A


 


3234




0.24
A


 


3235





A


 


3236





A


 


3237





A


 


3238





A


 


3239





A


 


3240





A


 


3241





A


 


3242




0.030
A


 


3243





A


 


3244




0.070
A


 


3245





A


 


3246





A


 


3247





A


 


3248





A


 


3249





A


 


3250





A


 


3251





A


 


3252





A


 


3253





A


 


3254





A


 


3255




0.029
A


 


3256





A


 


3257





A


 


3258





A


 


3259





A


 


3260





A


 


3261




0.072
A


 


3262





A


 


3263





A


 


3264




0.24
A


 


3265





B


 


3266





A


 


3267





A


 


3268





A


 


3269





A


 


3270





A


 


3271




1.38
A


 


3272





A


 


3273




0.47
A


 


3274





A


 


3275





A


 


3276





A


 


3277





A


 


3278





A


 


3279





A


 


3280





A


 


3281




1.29
A


 


3282





A


 


3283





A


 


3284





A


 


3285





A


 


3286





A


 


3287





A


 


3288





A


 


3289




2.53
A


 


3290





A


 


3291





A


 


3292





A


 


3293





A


 


3294





A


 


3295





A


 


3296




0.24
A


 


3297





A


 


3298





A


 


3299





A


 


3300





B


 


3301





A


 


3302





A


 


3303




5.59
A


 


3304





A


 


3305





A


 


3306





A


 


3307





A


 


3308





A


 


3309





A


 


3310




2.15
A


 


3311





A


 


3312





A


 


3313





A


 


3314





A


 


3315





A


 


3316





A


 


3317





A


 


3318





A


 


3319





A


 


3320




8.80
A


 


3321





B


 


3322





A


 


3323





A


 


3324





A


 


3325




2.40
A


 


3326





A


 


3327





A


 


3328





A


 


3329





A


 


3330





A


 


3331





A


 


3332





A


 


3333





A


 


3334




0.062
A


 


3335





A


 


3336




0.090
A


 


3337





A


 


3338





A


 


3339





A


 


3340





A


 


3341





A


 


3342





A


 


3343





A


 


3344





A


 


3345





A


 


3346




0.067
A


 


3347





A


 


3348





A


 


3349




0.041
A


 


3350





A


 


3351





A


 


3352





A


 


3353





A


 


3354





A


 


3355





A


 


3356





A


 


3357





A


 


3358




0.034
A


 


3359





A


 


3360





A


 


3361




0.16
A


 


3362





A


 


3363





A


 


3364





A


 


3365




0.024
A


 


3366




0.14
A


 


3367





A


 


3368





A


 


3369





A


 


3370





A


 


3371





A


 


3372





A


 


3373




0.052
A


 


3374





A


 


3375





A


 


3376





A


 


3377





A


 


3378





A


 


3379




0.14
A


 


3380





A


 


3381





A


 


3382





A


 


3383





A


 


3384





A


 


3385





A


 


3386





A


 


3387




0.068
A


 


3388




0.013
A


 


3389





A


 


3390





A


 


3391





A


 


3392





A


 


3393





A


 


3394





A


 


3395





A


 


3396





A


 


3397




0.082
A


 


3398





A


 


3399





A


 


3400





A


 


3401





A


 


3402




0.20
A


 


3403





A


 


3404





A


 


3405





A


 


3406




1.91
A


 


3407





A


 


3408





A


 


3409





A


 


3410





A


 


3411





A


 


3412





A


 


3413





B


 


3414





A


 


3415





A


 


3416





A


 


3417




0.032
A


 


3418





A


 


3419





A


 


3420





A


 


3421





A


 


3422





A


 


3423





A


 


3424





A


 


3425




0.033
A


 


3426





A


 


3427





A


 


3428





A


 


3429





A


 


3430




1.45
A


 


3431





A


 


3432





B


 


3433





B


 


3434





A


 


3435





A


 


3436




0.081
A


 


3437





A


 


3438





A


 


3439





A


 


3440





A


 


3441





A


 


3442





A


 


3443





A


 


3444





A


 


3445




0.061
A


 


3446





A


 


3447





A


 


3448




0.059
A


 


3449





A


 


3450





A


 


3451





A


 


3452





A


 


3453





A


 


3454





A


 


3455





A


 


3456





A


 


3457





A


 


3458





A


 


3459





A


 


3460




0.016
A


 


3461




0.027
A


 


3462





A


 


3463





A


 


3464





A


 


3465





A


 


3466





A


 


3467





A


 


3468





A


 


3469




0.035
A


 


3470





A


 


3471





A


 


3472





A


 


3473





A


 


3474





A


 


3475





A


 


3476





A


 


3477




0.12
A


 


3478





A


 


3479





A


 


3480





A


 


3481




0.065
A


 


3482





A


 


3483





A


 


3484





A


 


3485





A


 


3486




0.32
A


 


3487





A


 


3488





A


 


3489





A


 


3490




0.54
A


 


3491





B


 


3492




0.33
A


 


3493





A


 


3494





A


 


3495





A


 


3496





B


 


3497





A


 


3498





A


 


3499




0.46
A


 


3500





A


 


3501





A


 


3502





A


 


3503





A


 


3504





A


 


3505





A


 


3506




0.51
A


 


3507





A


 


3508





A


 


3509





A


 


3510





A


 


3511





A


 


3512





A


 


3513





A


 


3514





A


 


3515




0.017
A


 


3516





A


 


3517





A


 


3518




0.097
A


 


3519





A


 


3520





A


 


3521





A


 


3522





B


 


3523





A


 


3524





A


 


3525





B


 


3526





A


 


3527




0.21
A


 


3528





A


 


3529





A


 


3530





A


 


3531





A


 


3532





A


 


3533





A


 


3534




7.54
A


 


3535





A


 


3536





A


 


3537





A


 


3538





B


 


3539





B


 


3540




26.22
B


 


3541





B


 


3542





B


 


3543





A


 


3544





B


 


3545





A


 


3546





A


 


3547




2.09
A


 


3548





A


 


3549





A


 


3550





A


 


3551





A


 


3552





A


 


3553





A


 


3554




0.029
A


 


3555





A


 


3556





A


 


3557





A


 


3558




0.26
A


 


3559





A


 


3560





A


 


3561





A


 


3562




0.018
A


 


3563





A


 


3564





A


 


3565





A


 


3566





A


 


3567




0.035
A


 


3568





A


 


3569





A


 


3570





A


 


351 





A


 


3572





A


 


3573





A


 


3574





A


 


3575





A


 


3576





A


 


3577




0.16
A


 


3578





A


 


3579





A


 


3580





B


 


3581





A


 


3582




8.95
A


 


3583





A


 


3584





B


 


3585





A


 


3586





A


 


3587





A


 


3588





A


 


3589





A


 


3590




0.16
A


 


3591





A


 


3592





B


 


3593





A


 


3594





A


 


3595





A


 


3596





A


 


3597




3.05
A


 


3598





A


 


3599





A


 


3600





A


 


3601





A


 


3602





A


 


3603




0.80
A


 


3604





A


 


3605





A


 


3606





A


 


3607





A


 


3608





A


 


3609





A


 


3610





A


 


3611





A


 


3612





A


 


3613




0.046
A


 


3614





A


 


3615





A


 


3616





A


 


3617





A


 


3618





A


 


3619




0.084
A


 


3620





A


 


3621





A


 


3622





B


 


3623





A


 


3624





A


 


3625





A


 


3626





B


 


3627




0.10
A


 


3628





A


 


3629





A


 


3630





A


 


3631





A


 


3632





A


 


3633




0.058
A


 


3634





A


 


3635





A


 


3636





A


 


3637





A


 


3638





A


 


3639





A


 


3640




16.05
B


 


3641





A


 


3642





A


 


3643





B


 


3644





A


 


3645





A


 


3646





A


 


3647





A


 


3648




0.20
A


 


3649





A


 


3650





A


 


3651





A


 


3652




0.45
A


 


3653





A


 


3654





A


 


3655





A


 


3656





A


 


3657





A


 


3658





A


 


3659





A


 


3660




1.67
A


 


3661





A


 


3662





A


 


3663





A


 


3664




0.038
A


 


3665





A


 


3666





A


 


3667





A


 


3668





A


 


3669





A


 


3670





A


 


3671





A


 


3672





A


 


3673





A


 


3674




0.15
A


 


3675





A


 


3676





A


 


3677





A


 


3678





A


 


3679





A


 


3680





A


 


3681





A


 


3682




0.31
A


 


3683





A


 


3684





A


 


3685





A


 


3686





A


 


3687





A


 


3688




0.97
A


 


3689





A


 


3690





A


 


3691





A


 


3692





A


 


3693





A


 


3694




2.23
A


 


3695





A


 


3696





A


 


3697





A


 


3698





A


 


3699





A


 


3700





A


 


3701





A


 


3702





A


 


3703




1.33
A


 


3704





A


 


3705





A


 


3706





A


 


3707





A


 


3708





A


 


3709




0.71
A


 


3710





A


 


3711





A


 


3712





A


 


3713





A


 


3714





A


 


3715





A


 


3716




0.62
A


 


3717





A


 


3718





A


 


3719





A


 


3720





A


 


3721





A


 


3722





A


 


3723




1.34
A


 


3724





A


 


3725





A


 


3726





A


 


3727





A


 


3728





A


 


3729





A


 


3730




0.60
A


 


3731





A


 


3732





A


 


3733





A


 


3734





A


 


3735





A


 


3736





A


 


3737




1.63
A


 


3738





A


 


3739





A


 


3740





A


 


3741





A


 


3742





A


 


3743





A


 


3744




1.18
A


 


3745





A


 


3746





A


 


3747





A


 


3748





A


 


3749





A


 


3750





A


 


3751




0.14
A


 


3752





A


 


3753





A


 


3754





A


 


3755





A


 


3756





A


 


3757





A


 


3758




0.54
A


 


3759





A


 


3760





A


 


3761





A


 


3762





A


 


3763





A


 


3764





A


 


3765




0.68
A


 


3766





A


 


3767





A


 


3768





A


 


3769





A


 


3770





A


 


3771





A


 


3772




7.42
A


 


3773





A


 


3774





A


 


3775





A


 


3776





A


 


3777





A


 


3778





A


 


3779




0.46
A


 


3780





A


 


3781





A


 


3782





A


 


3783





A


 


3784





A


 


3785





A


 


3786




0.55
A


 


3787





A


 


3788





A


 


3789





A


 


3790





A


 


3791





A


 


3792





A


 


3793





A


 


3794




0.061
A


 


3795





A


 


3796





A


 


3797





A


 


3798





A


 


3799





A


 


3800




0.83
A


 


3801





A


 


3802





A


 


3803





A


 


3804





A


 


3805





A


 


3806





A


 


3807




0.28
A


 


3808





A


 


3809





A


 


3810





A


 


3811





A


 


3812




0.50
A


 


3813





A


 


3814





A


 


3815





A


 


3816





A


 


3817




0.11
A


 


3818





A


 


3819





A


 


3820





A


 


3821





A


 


3822





A


 


3823





A


 


3824




0.077
A


 


3825





A


 


3826





A


 


3827





A


 


3828




0.35
A


 


3829





A


 


3830





A


 


3831





A


 


3832





A


 


3833





A


 


3834




0.086
A


 


3835





A


 


3836





A


 


3837





A


 


3838




0.052
A


 


3839




0.078
A


 


3840





A


 


3841





A


 


3842





A


 


3843





A


 


3844





A


 


3845




0.022
A


 


3846





A


 


3847




0.027
A


 


3848





A


 


3849





A


 


3850





A


 


3851





A


 


3852





A


 


3853





A









Pharmaceutical Compositions and Methods of Treatment
The compounds demonstrate activity against HCV NS5B and can be useful in treating HCV and HCV infection. Therefore, another aspect of the invention is a composition comprising a compound, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
Another aspect of the invention is a composition further comprising a compound having anti-HCV activity.
Another aspect of the invention is a composition where the compound having anti-HCV activity is an interferon or a ribavirin. Another aspect of the invention is where the interferon is selected from interferon alpha 2B, pegylated interferon alpha, consensus interferon, interferon alpha 2A, interferon lambda, and lymphoblastoid interferon tau.
Another aspect of the invention is a composition where the compound having anti-HCV activity is a cyclosporin. Another aspect of the invention is where the cyclosporin is cyclosporin A.
Another aspect of the invention is a composition where the compound having anti-HCV activity is selected from the group consisting of interleukin 2, interleukin 6, interleukin 12, a compound that enhances the development of a type 1 helper T cell response, interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an inosine 5′-monophospate dehydrogenase inhibitor, amantadine, and rimantadine.
Another aspect of the invention is a composition where the compound having anti-HCV activity is effective to inhibit the function of a target selected from HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH, and a nucleoside analog for the treatment of an HCV infection.
Another aspect of the invention is a composition comprising a compound, or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable carrier, an interferon and ribavirin.
Another aspect of the invention is a method of inhibiting the function of the HCV replicon comprising contacting the HCV replicon with a compound or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a method of inhibiting the function of the HCV NS5B protein comprising contacting the HCV NS5B protein with a compound or a pharmaceutically acceptable salt thereof.
Another aspect of the invention is a method of treating an HCV infection in a patient comprising administering to the patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof. In another embodiment the compound is effective to inhibit the function of the HCV replicon. In another embodiment the compound is effective to inhibit the function of the HCV NS5B protein.
Another aspect of the invention is a method of treating an HCV infection in a patient comprising administering to the patient a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt thereof, in conjunction with (prior to, after, or concurrently) another compound having anti-HCV activity.
Another aspect of the invention is the method where the other compound having anti-HCV activity is an interferon or a ribavirin.
Another aspect of the invention is the method where the interferon is selected from interferon alpha 2B, pegylated interferon alpha, consensus interferon, interferon alpha 2A, interferon lambda, and lymphoblastoid interferon tau.
Another aspect of the invention is the method where the other compound having anti-HCV activity is a cyclosporin.
Another aspect of the invention is the method where the cyclosporin is cyclosporin A.
Another aspect of the invention is the method where the other compound having anti-HCV activity is selected from interleukin 2, interleukin 6, interleukin 12, a compound that enhances the development of a type 1 helper T cell response, interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an inosine 5′-monophospate dehydrogenase inhibitor, amantadine, and rimantadine.
Another aspect of the invention is the method where the other compound having anti-HCV activity is effective to inhibit the function of a target selected from the group consisting of HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH, and a nucleoside analog for the treatment of an HCV infection.
Another aspect of the invention is the method where the other compound having anti-HCV activity is effective to inhibit the function of target in the HCV life cycle other than the HCV NS5B protein.
“Therapeutically effective” means the amount of agent required to provide a meaningful patient benefit as understood by practitioners in the field of hepatitis and HCV infection.
“Patient” means a person infected with the HCV virus and suitable for therapy as understood by practitioners in the field of hepatitis and HCV infection.
“Treatment,” “therapy,” “regimen,” “HCV infection,” and related terms are used as understood by practitioners in the field of hepatitis and HCV infection.
The compounds of this invention are generally given as pharmaceutical compositions comprised of a therapeutically effective amount of a compound or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier and may contain conventional excipients. Pharmaceutically acceptable carriers are those conventionally known carriers having acceptable safety profiles. Compositions encompass all common solid and liquid forms including for example 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. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 17th edition, 1985.
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 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 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 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 hepatitis and HCV infection. In these combination methods, the compound 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.
Some examples of compounds suitable for compositions and methods are listed in Table 2.










TABLE 2







Type of Inhibitor or



Brand Name
Physiological Class
Target
Source Company







NIM811

Cyclophilin Inhibitor
Novartis


Zadaxin

Immuno-modulator
Sciclone


Suvus

Methylene blue
Bioenvision


Actilon

TLR9 agonist
Coley


(CPG10101)





Batabulin (T67)
Anticancer
β-tubulin inhibitor
Tularik Inc., South





San Francisco, CA


ISIS 14803
Antiviral
antisense
ISIS





Pharmaceuticals Inc,





Carlsbad, CA/Elan





Phamaceuticals Inc.,





New York, NY


Summetrel
Antiviral
antiviral
Endo





Pharmaceuticals





Holdings Inc.,





Chadds Ford, PA


GS-9132 (ACH-
Antiviral
HCV Inhibitor
Achillion/Gilead


806)





Pyrazolopyrimidine
Antiviral
HCV Inhibitors
Arrow Therapeutics


compounds and


Ltd.


salts





From WO-





2005047288





May 26, 2005





Levovirin
Antiviral
IMPDH inhibitor
Ribapharm Inc.,





Costa Mesa, CA


Merimepodib
Antiviral
IMPDH inhibitor
Vertex


(VX-497)


Pharmaceuticals





Inc., Cambridge,





MA


XTL-6865 (XTL-
Antiviral
monoclonal antibody
XTL


002)


Biopharmaceuticals





Ltd., Rehovot, Isreal


Telaprevir
Antiviral
NS3 serine protease
Vertex


(VX-950, LY-

inhibitor
Pharmaceuticals


570310)


Inc., Cambridge,





MA/Eli Lilly and





Co. Inc.,





Indianapolis, IN


HCV-796
Antiviral
NS5B Replicase
Wyeth/Viropharma




Inhibitor



NM-283
Antiviral
NS5B Replicase
Idenix/Novartis




Inhibitor



GL-59728
Antiviral
NS5B Replicase
Gene Labs/




Inhibitor
Novartis


GL-60667
Antiviral
NS5B Replicase
Gene Labs/




Inhibitor
Novartis


2′C MeA
Antiviral
NS5B Replicase
Gilead




Inhibitor



PSI 6130
Antiviral
NS5B Replicase
Roche




Inhibitor



R1626
Antiviral
NS5B Replicase
Roche




Inhibitor



2′C Methyl
Antiviral
NS5B Replicase
Merck


adenosine

Inhibitor



JTK-003
Antiviral
RdRp inhibitor
Japan Tobacco Inc.,





Tokyo, Japan


Levovirin
Antiviral
ribavirin
ICN





Pharmaceuticals,





Costa Mesa, CA


Ribavirin
Antiviral
ribavirin
Schering-Plough





Corporation,





Kenilworth, NJ


Viramidine
Antiviral
Ribavirin Prodrug
Ribapharm Inc.,





Costa Mesa, CA


Heptazyme
Antiviral
ribozyme
Ribozyme





Pharmaceuticals





Inc., Boulder, CO


BILN-2061
Antiviral
serine protease
Boehringer




inhibitor
Ingelheim Pharma





KG, Ingelheim,





Germany


SCH 503034
Antiviral
serine protease
Schering Plough




inhibitor



Zadazim
Immune modulator
Immune modulator
SciClone





Pharmaceuticals





Inc., San Mateo, CA


Ceplene
Immunomodulator
immune modulator
Maxim





Pharmaceuticals





Inc., San Diego, CA


CellCept
Immunosuppressant
HCV IgG immuno-
F. Hoffmann-La




suppressant
Roche LTD, Basel,





Switzerland


Civacir
Immunosuppressant
HCV IgG immuno-
Nabi




suppressant
Biopharmaceuticals





Inc., Boca Raton, FL


Albuferon-α
Interferon
albumin IFN-α2b
Human Genome





Sciences Inc.,





Rockville, MD


Infergen A
Interferon
IFN
InterMune




alfacon-1
Pharmaceuticals





Inc., Brisbane, CA


Omega IFN
Interferon
IFN-ω
Intarcia Therapeutics


IFN-β and EMZ701
Interferon
IFN-β and EMZ701
Transition





Therapeutics Inc.,





Ontario, Canada


Rebif
Interferon
IFN-β1a
Serono, Geneva,





Switzerland


Roferon A
Interferon
IFN-α2a
F. Hoffmann-La





Roche LTD, Basel,





Switzerland


Intron A
Interferon
IFN-α2b
Schering-Plough





Corporation,





Kenilworth, NJ


Intron A and
Interferon
IFN-α2b/α1-thymosin
RegeneRx


Zadaxin


Biopharma. Inc.,





Bethesda, MD/





SciClone





Pharmaceuticals Inc,





San Mateo, CA


Rebetron
Interferon
IFN-α2b/ribavirin
Schering-Plough





Corporation,





Kenilworth, NJ


Actimmune
Interferon
INF-γ
InterMune Inc.,





Brisbane, CA


Interferon-β
Interferon
Interferon-β-1a
Serono


Multiferon
Interferon
Long lasting IFN
Viragen/





Valentis


Wellferon
Interferon
Lympho-blastoid IFN-
GlaxoSmithKline




αn1
plc, Uxbridge, UK


Omniferon
Interferon
natural IFN-α
Viragen Inc.,





Plantation, FL


Pegasys
Interferon
PEGylated IFN-α2a
F. Hoffmann-La





Roche LTD, Basel,





Switzerland


Pegasys and
Interferon
PEGylated IFN-α2a/
Maxim


Ceplene

immune modulator
Pharmaceuticals





Inc., San Diego, CA


Pegasys and
Interferon
PEGylated IFN-
F. Hoffmann-La


Ribavirin

α2a/ribavirin
Roche LTD, Basel,





Switzerland


PEG-Intron
Interferon
PEGylated IFN-α2b
Schering-Plough





Corporation,





Kenilworth, NJ


PEG-Intron/
Interferon
PEGylated IFN-
Schering-Plough


Ribavirin

α2b/ribavirin
Corporation,





Kenilworth, NJ


IP-501
Liver protection
antifibrotic
Indevus





Pharmaceuticals





Inc., Lexington, MA


IDN-6556
Liver protection
caspase inhibitor
Idun





Pharmaceuticals





Inc., San Diego, CA


ITMN-191 (R-7227)
Antiviral
serine protease
InterMune




inhibitor
Pharmaceuticals





Inc., Brisbane, CA


GL-59728
Antiviral
NS5B Replicase
Genelabs




Inhibitor



ANA-971
Antiviral
TLR-7 agonist
Anadys


Boceprevir
Antiviral
serine protease
Schering Plough




inhibitor



TMS-435
Antiviral
serine protease
Tibotec BVBA,




inhibitor
Mechelen, Belgium


BI-201335
Antiviral
serine protease
Boehringer




inhibitor
Ingelheim Pharma





KG, Ingelheim,





Germany


MK-7009
Antiviral
serine protease
Merck




inhibitor



PF-00868554
Antiviral
replicase inhibitor
Pfizer


ANA598
Antiviral
Non-Nucleoside
Anadys




NS5B Polymerase
Pharmaceuticals,




Inhibitor
Inc., San Diego, CA,





USA


IDX375
Antiviral
Non-Nucleoside
Idenix




Replicase Inhibitor
Pharmaceuticals,





Cambridge, MA,





USA


BILB 1941
Antiviral
NS5B Polymerase
Boehringer




Inhibitor
Ingelheim Canada





Ltd R&D, Laval,





QC, Canada


PSI-7851
Antiviral
Nucleoside
Pharmasset,




Polymerase Inhibitor
Princeton, NJ, USA


PSI-7977
Antiviral
Nucleotide NS5B
Pharmasset,




Polymerase Inhibitor
Princeton, NJ, USA


VCH-759
Antiviral
NS5B Polymerase
ViroChem Pharma




Inhibitor



VCH-916
Antiviral
NS5B Polymerase
ViroChem Pharma




Inhibitor



GS-9190
Antiviral
NS5B Polymerase
Gilead




Inhibitor



Peg-interferon
Antiviral
Interferon
ZymoGenetics/Bristol-


lamda


Myers Squibb









Synthetic Methods
The compounds may be made by methods known in the art including those described below and including variations within the skill of the art. Some reagents and intermediates are known in the art. Other reagents and intermediates can be made by methods known in the art using readily available materials. The variables (e.g. numbered “R” substituents) used to describe the synthesis of the compounds are intended only to illustrate how to make the compounds and are not to be confused with variables used in the claims or in other sections of the specification. The following methods are for illustrative purposes and are not intended to limit the scope of the invention.
Abbreviations used in the schemes generally follow conventions used in the art. Chemical abbreviations used in the specification and examples are defined as follows: “NaHMDS” for sodium bis(trimethylsilyl)amide; “DMF” for N,N-dimethylformamide; “MeOH” for methanol; “NBS” for N-bromosuccinimide; “Ar” for aryl; “TFA” for trifluoroacetic acid; “LAH” for lithium aluminum hydride; “BOC”, “DMSO” for dimethylsulfoxide; “h” for hours; “rt” for room temperature or retention time (context will dictate); “min” for minutes; “EtOAc” for ethyl acetate; “THF” for tetrahydrofuran; “EDTA” for ethylenediaminetetraacetic acid; “Et2O” for diethyl ether; “DMAP” for 4-dimethylaminopyridine; “DCE” for 1,2-dichloroethane; “ACN” for acetonitrile; “DME” for 1,2-dimethoxyethane; “HOBt” for 1-hydroxybenzotriazole hydrate; “DIEA” for diisopropylethylamine, “Nf” for CF3(CF2)3SO2—; and “TMOF” for trimethylorthoformate.
Abbreviations are defined as follows: “1×” for once, “2×” for twice, “3×” for thrice, “° C.” for degrees Celsius, “eq” for equivalent or equivalents, “g” for gram or grams, “mg” for milligram or milligrams, “L” for liter or liters, “mL” for milliliter or milliliters, “μL” for microliter or microliters, “N” for normal, “M” for molar, “mmol” for millimole or millimoles, “min” for minute or minutes, “h” for hour or hours, “rt” for room temperature, “RT” for retention time, “atm” for atmosphere, “psi” for pounds per square inch, “conc.” for concentrate, “sat” or “sat'd” for saturated, “MW” for molecular weight, “mp” for melting point, “ee” for enantiomeric excess, “MS” or “Mass Spec” for mass spectrometry, “ESI” for electrospray ionization mass spectroscopy, “HR” for high resolution, “HRMS” for high resolution mass spectrometry, “LCMS” for liquid chromatography mass spectrometry, “HPLC” for high pressure liquid chromatography, “RP HPLC” for reverse phase HPLC, “TLC” or “tlc” for thin layer chromatography, “NMR” for nuclear magnetic resonance spectroscopy, “1H” for proton, “δ” for delta, “s” for singlet, “d” for doublet, “t” for triplet, “q” for quartet, “m” for multiplet, “br” for broad, “Hz” for hertz, and “α”, “β”, “R”, “S”, “E”, and “Z” are stereochemical designations familiar to one skilled in the art.
LC/MS Method (i.e., Compound Identification).
All Liquid Chromatography (LC) data were recorded on a Shimadzu LC-10AS or LC-20AS liquid chromotograph using a SPD-10AV or SPD-20A UV-Vis detector and Mass Spectrometry (MS) data were determined with a Micromass Platform for LC in electrospray mode.
HPLC Method (i.e., Compound Isolation).
Compounds purified by preparative HPLC were diluted in methanol (1.2 mL) and purified using a Shimadzu LC-8A or LC-10A or Dionex APS-3000 or Waters AcquityTM automated preparative HPLC system.
Syntheses of Intermediates
Preparation of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid, In-1001




Step 1:
To a solution of 2,4,6-trichloro-1,3,5-triazine (15 g) in THF (300 mL) was added 2,2,2-trifluoroethanol (8.14 g) and Hunig'sBase (15.63 mL). The resulting mixture was stirred for 16 hours. After removal of most THF and precipitate through a plug washing with THF, the filtrate was concentrate to give a crude that will be used as it is.
Step 2:
To a solution of the product in Step 1 above (10 g) in THF (100 mL) was added tert-butyl 4-aminobenzoate (7.79 g) and Hunig'sBase (7.04 mL). The resulting mixture was stirred for 16 h. The precipitate was filtered and washed with Et2O, dried, then washed with water and dried to give 10.6 g of tert-butyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate as a solid.










tert-butyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-ylamino)benzoate


















MS (M + H)+ Calcd.
405.1



MS (M + H)+ Observ.
405.0







LC Condition










Solvent A
100% Water-0.1% TFA



Solvent B
100% ACN-0.1% TFA



Start % B
 2



Final % B
 98



Gradient Time
1.6 min



Stop Time
1.8 min



Flow Rate
0.8 mL/min



Wavelength
220



Solvent Pair
ACN-H2O-0.1% TFA



Column
Aquity UPLC BEH C18 1.7 um










Step 3:
To a slurry of tert-butyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate (3.6 g) and 1-(4-chlorophenyl)cyclopropanamine (1.49 g) in THF (50 mL) was stirred for 5 hours at 80° C. The precipitate was filtrated through a plug washing with THF to give acrude product that was purified by Biotage eluting with 4/1-hexane/ethyl acetate to give 1.8 g of tert-butyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate as a solid.










tert-butyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate
















MS (M + H)+ Calcd.
536.2


MS (M + H)+ Observ.
536.0







LC Condition








Solvent A
100% Water-0.1% TFA


Solvent B
100% ACN-0.1% TFA


Start % B
 2


Final % B
 98


Gradient Time
1.6 min


Stop Time
1.8 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
ACN-H2O-0.1% TFA


Column
Aquity UPLC BEH C18 1.7 um









Step 4:
A solution of above tert-butyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate (4 g) and HCl in dioxane (7.46 ml, 4M) was stirred for 4 hours. Concentration gave 3.58 g of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid as a solid.










4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-ylamino)benzoic acid, In-1001
















MS (M + H)+ Calcd.
480.1


MS (M + H)+ Observ.
480.1







LC Condition








Solvent A
100% Water-0.1% TFA


Solvent B
100% ACN-0.1% TFA


Start % B
 2


Final % B
 98


Gradient Time
1.6 min


Stop Time
1.8 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
ACN-H2O-0.1% TFA


Column
Aquity UPLC BEH C18 1.7 um









Preparation of (S)-methyl 3-amino-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoate, In-1002




Step 1:
To a solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid (50 mg) in DMF (2 mL) was added O-(benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate (23.82 mg) and (S)-methyl 2-amino-3-(tert-butoxycarbonylamino)propanoate hydrochloride (18.90 mg) and iPr2NEt (0.052 ml). After stirring at rt for 4 h, the mixture was purified by preparative HPLC to give (S)-methyl 3-(tert-butoxycarbonylamino)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoate.










(S)-methyl 3-(tert-butoxycarbonylamino)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-


ylamino)benzamido)propanoate
















MS (M + H)+ Calcd.
680.2


MS (M + H)+ Observ.
680.3


Retention Time
3.44 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 50 × 2, 3 u









Step 2:
To a solution of (S)-methyl 3-(tert-butoxycarbonylamino)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoate (1 g) in CH2Cl2 (10 mL) was added TFA (3 mL). The mixture was stirred at room temperature for 16 hours. All the solvents were removed under vacuum to give (S)-methyl 3-amino-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoate (0.8 g).










(S)-methyl 3-amino-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoate, In-1002
















MS (M + H)+ Calcd. 
580.2


MS (M + H)+ Observ.
580.0


Retention Time
1.35 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









Preparation of (S)-methyl 4-amino-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)butanoate, In-1003




In-1003 was prepared by using the same procedure of synthesizing In-1002, using (S)-methyl 2-amino-4-(tert-butoxycarbonylamino)butanoate as a starting material.










(S)-methyl 4-amino-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)butanoate:
















MS (M + H)+ Calcd.
594.2


MS (M + H)+ Observ.
594.4


Retention Time
1.64 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex LUNA C18, 30 × 2, 3u









Preparation of (S)-methyl 5-amino-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)pentanoate




In-1004 was prepared by using the same procedure of synthesizing In-1002, using (S)-methyl 2-amino-5-(tert-butoxycarbonylamino)pentanoate as a starting material.










(S)-methyl 5-amino-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)pentanoate, In-1004
















MS (M + H)+ Calcd.
608.2


MS (M + H)+ Observ.
608.1


Retention Time
2.44







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Preparation of N-(3-amino-2,2-dimethylpropyl)-4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamide, In-1005




iPr2NEt (0.266 g) and O-benzotriazol-1-yl-N,N,N′,N′-tetra-methyluronium tetrafluoroborate (0.610 g) were added into a solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid (0.760 g) and tert-butyl 3-amino-2,2-dimethylpropylcarbamate (0.352 g) were dissolved in DMF (3 mL). The reaction was stirred at room temperature for 16 hrs before 200 mL of EtOAc was added. The organic phase was washed with water (2×25 mL) and brine (20 mL), dried over MgSO4, and concentrated to give crude product tert-butyl 3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2,2-dimethylpropylcarbamate (0.75 g) which was dissolved in CH2Cl2 (3 mL) and TFA (1.3 mL). After being stirred at room temperature for 16 hours, the mixture was concentrated to give a crude product, N-(3-amino-2,2-dimethylpropyl)-4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamide (0.63 g).










tert-butyl 3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-


6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-


benzamido)-2,2-dimethylpropylcarbamate
















MS (M + H)+ Calcd.
664.3


MS (M + H)+ Observ.
664.4


Retention Time
3.75 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM 



Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM 



Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex LUNA C18, 50 × 2, 3 u



















N-(3-amino-2,2-dimethylpropy1)-4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-ylamino)benzamide, In-1005
















MS (M + H)+ Calcd.
564.2


MS (M + H)+ Observ.
564.3


Retention Time
3.02 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM 



Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM 



Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex LUNA C18, 50 × 2, 3 u









Preparation of N-(3-amino-2,2-dimethylpropyl)-4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamide, In-1006
Intermediate In-1006 was prepared by using the same synthetic route of synthesizing In-1005, using methyl 4-amino-2-fluorobenzoate as one of the starting materials.












In-1006






















MS (M + H)+ Calcd.
582.2


MS (M + H)+ Observ.
582.2


Retention Time
1.74 min







LC Condition








Solvent A
90% Water-10% Methanol-0.l% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









Synthesis of Compound 1001, (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-ethoxy-2-oxoacetamido)propanoate




To a solution of In-1002 (25 mg) in CH2Cl2 (5 mL) was added ethyl 2-chloro-2-oxoacetate (5.89 mg). The mixture was stirred at room temperature for 4 hours. The reaction was quenched with the MeOH and all the solvents were removed under vacuum. The residue was purified by preparative HPLC to give 1001 (5 mg).










(S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-ethoxy-2-


oxoacetamido)propanoate, 1001
















MS (M + H)+ Calcd.
680.2


MS (M + H)+ Observ.
680.1


Retention Time
3.16 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1002, (S)-methyl 3-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoate




To a solution of 2-amino-2-oxoacetic acid (11.52 mg) and TBTU (41.5 mg) in DMF (2 mL) was added In-1002 (50 mg). After stirring at room temperature for 2 hours, the mixture was purified by preparative HPLC to give 1002 (14 mg).










(S)-methyl 3-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-ylamino)benzamido)propanoate, 1002
















MS (M + H)+ Calcd.
651.2


MS (M + H)+ Observ.
651.0


Retention Time
3.61 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 40


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1003, (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(methylamino)-2-oxoacetamido)propanoate




To a solution of 2-(methylamino)-2-oxoacetic acid (13.33 mg) and TBTU (41.5 mg) in DMF (Volume: 2 mL) was added In-1002 (50 mg). After stirring at room temperature for 2 hours, the mixture was purified by preparative HPLC to give 1003 (20 mg).










(S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-


(2-(methylamino)-2-oxoacetamido)propanoate, 1003
















MS (M + H)+ Calcd.
665.2


MS (M + H)+ Observ.
665.1


Retention Time
4.26 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1004, (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(dimethylamino)-2-oxoacetamido)propanoate




To a solution of 2-(dimethylamino)-2-oxoacetic acid (15.14 mg) and TBTU (41.5 mg) in DMF (2 mL) was added In-1002 (50 mgl). After stirring at room temperature for 2 hours, the mixture was purified by preparative HPLC to give 1004 (28 mg).










(S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-


(2-(dimethylamino)-2-oxoacetamido)propanoate, 1004
















MS (M + H)+ Calcd.
679.2


MS (M + H)+ Observ.
679.1


Retention Time
3.61 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 40


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1005, (S)-3-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoic acid




A suspension of 1002 (14 mg) and K2CO3 (8.92 mg) in acetone-water (1:1, 4 mL) was stirred at room temperature for 16 hours. All the solvents were removed under vacuum. The residue was purified by preparative HPLC to give 1005 (12.6 mg).










(S)-3-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)-


cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-ylamino)benzamido)propanoic acid, 1005
















MS (M + H)+ Calcd.
637.2


MS (M + H)+ Observ.
637.0


Retention Time
2.83 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 60


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1006, (S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(methylamino)-2-oxoacetamido)propanoic acid




A suspension of 1003 (12 mg) and K2CO3 (7.48 mg) in acetone-water (1:1, 4 mL) was stirred at room temperature for 16 hours. All the solvents were removed under vacuum. The residue was purified by preparative HPLC to give 1006 (8.4 mg).










(S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


3-(2-(methylamino)-2-oxoacetamido)propanoic acid, 1006
















MS (M + H)+ Calcd.
651.2


MS (M + H)+ Observ.
651.1


Retention Time
3.01 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 60


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1007, (S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(dimethylamino)-2-oxoacetamido)propanoic acid




A suspension of 1004 (20 mg) and K2CO3 (12.2 mg) in acetone-water (1:1, 4 mL) was stirred at room temperature for 16 hours. All the solvents were removed under vacuum. The residue was purified by preparative HPLC to give 1007 (7 mg).










(S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


3-(2-(dimethylamino)-2-oxoacetamido)propanoic acid, 1007
















MS (M + H)+ Calcd.
665.2


MS (M + H)+ Observ.
665.1


Retention Time
3.10 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 60


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1008, N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2,2-dimethylpropyl)-N2,N2-dimethyloxalamide




iPr2NEt (0.037 g) and O-benzotriazol-1-yl-N,N,N′,N′-tetra-methyluronium tetrafluoroborate (0.068 g) was added into the solution of crude In-1005 (0.080 g) and 2-(dimethylamino)-2-oxoacetic acid (0.017 g) in DMF (2 mL). The reaction was stirred at room temperature for 6 hours. All the solvents were removed under vacuum. The residue was purified by preparative HPLC to give 1008 (40 mg).










N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


2,2-dimethylpropy1)-N2,N2-dimethyloxalamide, 1008
















MS (M + H)+ Calcd.
663.2


MS (M + H)+ Observ.
663.1


Retention Time
2.00 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
5 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 4.6 × 50 mm S10









Synthesis of Compound 1479, N1-(3-(4-((4-(1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-2,2-dimethylpropyl)-N2-(4-fluorophenyl)oxalamide
Compound 1479 was prepared by using the same procedure of synthesizing 1008, using 2-((4-fluorophenyl)amino)-2-oxoacetic acid as one of the starting materials. Its LCMS is shown in a later section.




The following selected examples were prepared by using the same procedure of synthesizing 1479, using the corresponding mono-Boc diamines rather than mono-Boc 2,2-dimethylpropane-1,3-diamine as starting materials:












3335

















MS (M + H)+ Calcd.
715.2


MS (M + H)+ Observ.
715.1


Retention Time
4.48 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um










3360

















MS (M + H)+ Calcd.
701.2


MS (M + H)+ Observ.
701.0


Retention Time
4.42 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um










3365

















MS (M + H)+ Calcd.
717.2


MS (M + H)+ Observ.
717.2


Retention Time
4.42 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um










3373

















MS (M + H)+ Calcd.
687.2


MS (M + H)+ Observ.
687.0


Retention Time
4.40 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um










3395

















MS (M + H)+ Calcd.
715.2


MS (M + H)+ Observ.
715.3


Retention Time
1.89 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u










3412

















MS (M + H)+ Calcd.
729.2


MS (M + H)+ Observ.
729.3


Retention Time
1.87 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u










3418

















MS (M + H)+ Calcd.
715.2


MS (M + H)+ Observ.
715.3


Retention Time
2.01 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u










3448

















MS (M + H)+ Calcd.
715.2


MS (M + H)+ Observ.
715.5


Retention Time
2.36 min







LC Condition








Solvent A
90% Water -10% Methanol-0.1% TFA


Solvent B
10% Water -90% Methanol-0.1% TFA


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um










3482

















MS (M + H)+ Calcd.
741.2


MS (M + H)+ Observ.
741.3


Retention Time
1.89 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u










3499

















MS (M + H)+ Calcd.
745.2


MS (M + H)+ Observ.
745.3


Retention Time
3.18 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 50 × 2, 3 u









Synthesis of Compound 1009, (S)-methyl 4-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)butanoate




To a solution of In-1003 (50 mg) and 2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouronium tetrafluoroborate (54 mg) in DMF (1.5 mL) was added 2-amino-2-oxoacetic acid (15 mg). After stirring at room temperature for 4 hours, the mixture was purified by preparative HPLC to give 1009 (23 mg).










(S)-methyl 4-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-ylamino)benzamido)butanoate, 1009
















MS (M + H)+ Calcd.
665.2


MS (M + H)+ Observ.
665.1


Retention Time
3.61 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1010, (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-(2-(methylamino)-2-oxoacetamido)butanoate




1010 was prepared by using the same procedure of synthesizing 1009, using 2-(methylamino)-2-oxoacetic acid as a starting material.










(S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-


(2-(methylamino)-2-oxoacetamido)butanoate, 1010
















MS (M + H)+ Calcd.
679.2


MS (M + H)+ Observ.
679.1


Retention Time
3.57 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1011, (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-(2-(dimethylamino)-2-oxoacetamido)butanoate




1011 was prepared by using the same procedure of synthesizing 1009, using 2-(dimethylamino)-2-oxoacetic acid as a starting material.










(S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-


(2-(dimethylamino)-2-oxoacetamido)butanoate, 1011
















MS (M + H)+ Calcd.
693.2


MS (M + H)+ Observ.
693.1


Retention Time
3.53 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1012, (S)-methyl 5-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)pentanoate




1012 was prepared by using the same procedure of synthesizing 1009, using In-1004 as a starting material.










(S)-methyl 5-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-


ylamino)benzamido)pentanoate, 1012
















MS (M + H)+ Calcd.
679.2


MS (M + H)+ Observ.
679.1


Retention Time
3.64 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1013, (S)-4-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)butanoic acid




A mixture of 1009 (21 mg) and K2CO3 (18 mg) in acetone (2 mL)/water (2 mL) was stirred at room temperature for 16 hours. All the solvents were removed under vacuum. The residue was dissolved in MeOH and purified by preparative HPLC to give 1013 (7.5 mg).










(S)-4-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)-


cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)


benzamido)butanoic acid, 1013
















MS (M + H)+ Calcd.
651.2


MS (M + H)+ Observ.
651.1


Retention Time
3.53 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1014, (S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-(2-(methylamino)-2-oxoacetamido)butanoic acid




1014 was prepared by using the same procedure of synthesizing 1013, using 1010 as a starting material.










(S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-


(2-(methylamino)-2-oxoacetamido)butanoic acid, 1014
















MS (M + H)+ Calcd.
665.2


MS (M + H)+ Observ.
665.2


Retention Time
3.47 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1015, (S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-(2-(dimethylamino)-2-oxoacetamido)butanoic acid




1015 was prepared by using the same procedure of synthesizing 1013, using 1011 as a starting material.










(S)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-


(2-(dimethylamino)-2-oxoacetamido)butanoic acid, 1015
















MS (M + H)+ Calcd.
679.2


MS (M + H)+ Observ.
679.2


Retention Time
3.41 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1016, (S)-5-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)pentanoic acid




1016 was prepared by using the same procedure of synthesizing 1013, using 1012 as a starting material.










(S)-5-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)-


cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-


ylamino)benzamido)pentanoic acid,1016
















MS (M + H)+ Calcd.
665.2


MS (M + H)+ Observ.
665.1


Retention Time
3.48 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1017, methyl 2-(1-(2-amino-2-oxoacetyl)piperidin-3-yl)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)acetate, and 1018, 2-(1-(2-amino-2-oxoacetyl)piperidin-3-yl)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)acetic acid, and 1019, N-(1-(1-(2-amino-2-oxoacetyl)piperidin-3-yl)-2-oxo-2-(propylamino)ethyl)-4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamide







Step 1: To a solution of In-1001 (100 mg) and TBTU (100 mg) in DMF (6 mL) was added tert-butyl 3-(1-amino-2-methoxy-2-oxoethyl)piperidine-1-carboxylate (68.1 mg) and DIPEA (0.146 mL). After stirring at room temperature for 4 hours, the mixture was purified by preparative HPLC to give tert-butyl 3-(1-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2-methoxy-2-oxoethyl)piperidine-1-carboxylate (90 mg).










tert-Butyl 3-(1-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2-methoxy-2-


oxoethyl)piperidine-1-carboxylate
















MS (M + H)+ Calcd.
734.2


MS (M + H)+ Observ.
734.1


Retention Time
4.00 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 2: To a solution of tert-butyl 3-(1-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2-methoxy-2-oxoethyl)piperidine-1-carboxylate (85 mg) in DCM (5 mL) was added TFA (0.357 mL). The mixture was stirred at room temperature for 4 hours. All the solvents were removed under vacuum to give methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2-(piperidin-3-yl)acetate (70 mg).










Methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


2-(piperidin-3-yl)acetate
















MS (M + H)+ Calcd.
634.2


MS (M + H)+ Observ.
634.1


Retention Time
2.60 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 3: To a solution of 2-amino-2-oxoacetic acid (18.26 mg) and TBTU (65.8 mg) in DMF (2 mL) was added methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2-(piperidin-3-yl)acetate (65 mg). After stirring at room temperature for 2 hours, the mixture was purified by preparative HPLC to give 1017 (35 mg).










methyl 2-(1-(2-amino-2-oxoacetyl)piperidin-3-y1)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-ylamino)benzamido)acetate, 1017
















MS (M + H)+ Calcd.
705.2


MS (M + H)+ Observ.
705.1


Retention Time
3.72 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 4: A mixture of 1017 (35 mg) and K2CO3 (21 mg) in acetone (4 mL)/water (4.00 mL) was stirred at room temperature for 16 hours. All the solvents were removed under vacuum. The residue was purified by preparative HPLC to give 1018 (28 mg).










2-(1-(2-amino-2-oxoacetyl)piperidin-3-yl)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-ylamino)benzamido)acetic acid, 1018
















MS (M + H)+ Calcd.
691.2


MS (M + H)+ Observ.
691.1


Retention Time
3.74 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 5: To a solution of 1018 (15 mg) and TBTU (10.5 mg) in DMF (1.5 mL) was added propan-1-amine (6.42 mg). After stirring at room temperature for 16 hours, the mixture was diluted with water (200 mL) and extracted with EtOAc (2×300 mL). The organic layers were combined, washed with water (200 mL), brine (200 mL), dried over MgSO4 and concentrated. The residue was purified by silica gel column to give 1019 (1.1 mg).










N-(1-(1-(2-amino-2-oxoacetyl)piperidin-3-yl)-2-oxo-2-


(propylamino)ethyl)-4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamide, 1019
















MS (M + Na)+ Calcd.
754.2


MS (M + Na)+ Observ.
754.2


Retention Time
3.65 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1020, (S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(dimethylamino)-2-oxoacetamido)propanoate




Step 1: A mixture of In-1002 (70 mg), K2CO3 (33.4 mg) and ethanol (2 mL) was stirred at room temperature for three days. The mixture was acidified by 1N HCl, diluted by MeOH and purified by preparative HPLC to give In-1101 (30 mg).










(S)-ethyl 3-amino-2-(4-(4-(1-(4-chlorophenyl)-


cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-


2-ylamino)benzamido)propanoate
















MS (M + Na)+ Calcd.
594.2


MS (M + Na)+ Observ.
594.2


Retention Time
3.38 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 2: To a solution of 2-(dimethylamino)-2-oxoacetic acid (7.6 mg) and TBTU (20.9 mg) in DMF (1 mL) was added In-1101 (30 mg) and DIPEA (0.03 mL). After stirring at room temperature for 2 hours, the mixture was purified by preparative HPLC to give 1020 (15.0 mg).










(S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-


(2-(dimethylamino)-2-oxoacetamido)propanoate, 1020
















MS (M + H)+ Calcd.
693.2


MS (M + H)+ Observ.
693.2


Retention Time
3.89 min


 







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1021, (S)-isopropyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(dimethylamino)-2-oxoacetamido)propanoate




1021 was prepared by using the same procedure of synthesizing 1020, using iPrOH as a starting material.










(S)-isopropyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


3-(2-(dimethylamino)-2-oxoacetamido)propanoate, 1021
















MS (M + H)+ Calcd.
707.2


MS (M + H)+ Observ.
707.2


Retention Time
4.03 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1022, (S)-2-morpholinoethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(dimethylamino)-2-oxoacetamido)propanoate




1022 was prepared by using the same procedure of synthesizing 1020, using 2-morpholinoethanol as a starting material.










(S)-2-morpholinoethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-


6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-


(2-(dimethylamino)-2-oxoacetamido)propanoate, 1022
















MS (M + H)+ Calcd.
778.3


MS (M + H)+ Observ.
778.3


Retention Time
3.22 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1023, (S)-2-(2-hydroxyethoxy)ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(dimethylamino)-2-oxoacetamido)propanoate




1023 was prepared by using the same procedure of synthesizing 1020, using 2-morpholinoethanol as a starting material.










(S)-2-(2-hydroxyethoxy)ethyl 2-(4-(4-(1-(4-chlorophenyl)-


cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-


benzamido)-3-(2-(dimethylamino)-2-oxoacetamido)propanoate, 1023
















MS (M + H)+ Calcd.
753.2


MS (M + H)+ Observ.
753.3


Retention Time
3.64 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1024, (S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-(methylamino)-2-oxoacetamido)propanoate




1024 was prepared by using the same procedure of synthesizing 1020, using 2-(methylamino)-2-oxoacetic acid as a starting material.










((S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-


(2-(methylamino)-2-oxoacetamido)propanoate, 1024
















MS (M + H)+ Calcd.
679.2


MS (M + H)+ Observ.
679.1


Retention Time
3.93 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1025, (S)-ethyl 3-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)propanoate




1025 was prepared by using the same procedure of synthesizing 1020, using 2-amino-2-oxoacetic acid as a starting material.










(S)-ethyl 3-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-ylamino)benzamido)propanoate, 1025
















MS (M + H)+ Calcd.
665.2


MS (M + H)+ Observ.
665.1


Retention Time
3.78 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water -Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1552, N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-2,2-dimethylpropyl)oxalamide




1552 was prepared by using the same procedure of synthesizing 1025, using In-1005 instead of In-1101.










Compound 1552


















MS (M + H)+ Calcd.
635.2



MS (M + H)+ Observ.
635.5



Retention Time
1.83 min







LC Condition










Solvent A
5% ACN: 95% Water: 10 mM 




Ammonium Actetate



Solvent B
95% ACN: 5% Water: 10 mM 




Ammonium Actetate



Start % B
 0



Final % B
100



Gradient Time
2 min



Flow Rate
1 mL/min



Wavelength
220



Solvent Pair
ACN: Water: Ammonium Actetate



Column
Phenomenex Luna C18, 30 × 2, 3u










Synthesis of Compound 1026, (S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-(2-(dimethylamino)-2-oxoacetamido)butanoate




To a solution of 1011 (20 mg) in ethanol (2 mL) was added K2CO3 (20 mg). The mixture was stirred at room temperature for 2 days and then purified by preparative HPLC to give 1026 (5.2 mg).










(S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-


(2-(dimethylamino)-2-oxoacetamido)butanoate, 1026
















MS (M + H)+ Calcd. 
707.2


MS (M + H)+ Observ.
707.1


Retention Time
3.60 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1027, (S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-(2-(methylamino)-2-oxoacetamido)butanoate




1027 was prepared by using the same procedure of synthesizing 1026, using 1010 as a starting material.










(S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-


(2-(methylamino)-2-oxoacetamido)butanoate, 1027
















MS (M + H)+ Calcd.
693.2


MS (M + H)+ Observ.
693.1


Retention Time
3.72 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1028, (S)-ethyl 4-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)butanoate




1028 was prepared by using the same procedure of synthesizing 1026, using 1009 as a starting material.










(S)-ethyl 4-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-ylamino)benzamido)butanoate, 1028
















MS (M + H)+ Calcd. 
679.2


MS (M + H)+ Observ.
679.1


Retention Time
3.56 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1029, (S)-ethyl 5-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)pentanoate




1029 was prepared by using the same procedure of synthesizing 1026, using 1012 as a starting material.










(S)-ethyl 5-(2-amino-2-oxoacetamido)-2-(4-(4-(1-(4-


chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-ylamino)benzamido)pentanoate, 1029
















MS (M + H)+ Calcd. 
693.2


MS (M + H)+ Observ.
693.1


Retention Time
3.54 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1030, (S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-5-(2-(methylamino)-2-oxoacetamido)pentanoate




Step 1: To a solution of 2-(methylamino)-2-oxoacetic acid (16.1 mg) and TBTU (50.1 mg) in DMF (1 mL) was added In-1004 (100 mg) and DIPEA (0.15 mL). After stirring at room temperature for 2 hours, the mixture was purified by preparative HPLC to give (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-5-(2-(methylamino)-2-oxoacetamido)pentanoate (58 mg).










(S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-5-


(2-(methylamino)-2-oxoacetamido)pentanoate
















MS (M + H)+ Calcd.
693.2


MS (M + H)+ Observ.
693.1


Retention Time
3.54 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 2: 1030 was prepared by using the same procedure of synthesizing 1026, using (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-5-(2-(methylamino)-2-oxoacetamido)pentanoate as a starting material.










(S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-5-


(2-(methylamino)-2-oxoacetamido)pentanoate, 1030
















MS (M + H)+ Calcd. 
707.2


MS (M + H)+ Observ.
707.1


Retention Time
3.68 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1031, (S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-5-(2-(dimethylamino)-2-oxoacetamido)pentanoate




Step 1: To a solution of 2-(dimethylamino)-2-oxoacetic acid (18.3 mg) and TBTU (50 mg) in DMF (1 mL) was added In-1004 (100 mg) and DIPEA (0.15 mL). After stirring at room temperature for 2 hours, the mixture was purified by preparative HPLC to give (S)-methyl 2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-5-(2-(dimethylamino)-2-oxoacetamido)pentanoate (48 mg).










(S)-methyl 2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-5-


(2-(dimethylamino)-2-oxoacetamido)pentanoate
















MS (M + H)+ Calcd. 
707.2


MS (M + H)+ Observ.
707.1


Retention Time
3.44 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 2: 1031 was prepared by using the same procedure of synthesizing 1026, using (S)-methyl 2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-5-(2-(dimethylamino)-2-oxoacetamido)pentanoate as a starting material.










(S)-ethyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-5-


(2-(dimethylamino)-2-oxoacetamido)pentanoate, 1031
















MS (M + H)+ Calcd. 
721.2


MS (M + H)+ Observ.
721.1


Retention Time
3.70 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Synthesis of Compound 1032, cyclopropylmethyl 2-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2,2-dimethylpropylamino)-2-oxoacetate




A mixture of In-1005 (50 mg) and diphenyl oxalate (100 mg) in THF (5 mL) was stirred at room temperature for 16 hours, before cyclopropylmethanamine hydrochloride (191 mg) and iPr2NEt (0.5 mL) were added. The reaction was carried out at room temperature for 4 hours. Removal of solvents gave a residue which was purified by preparative HPLC to offer 1032 (20.7 mg).










cyclopropylmethyl 2-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-


6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


2,2-dimethylpropylamino)-2-oxoacetate, 1032
















MS (M + Na)+ Calcd. 
712.2


MS (M + Na)+ Observ.
712.2


Retention Time
2.38 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









Synthesis of Compound 1033, N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2,2-dimethylpropyl)-N2-cycloheptyloxalamide




1033 was prepared by using the same procedure of synthesizing 1032, using cycloheptanamine as a starting material.










N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


2,2-dimethylpropyl)-N2-cycloheptyloxalamide, 1033
















MS (M + H)+ Calcd.
731.3


MS (M + H)+ Observ.
731.4


Retention Time
2.53 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









Synthesis of Compound 1034, N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2,2-dimethylpropyl)-N2-(2,4-difluorobenzyl)oxalamide




1034 was prepared by using the same procedure of synthesizing 1032, using (2,4-difluorophenyl)methanamine as a starting material.










N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


2,2-dimethylpropyl)-N2-(2,4-difluorobenzyl)oxalamide, 1034
















MS (M + H)+ Calcd.
761.2


MS (M + H)+ Observ.
761.3


Retention Time
2.47 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









Synthesis of Compound 1035, N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2,2-dimethylpropyl)-N2-phenyloxalamide




1035 was prepared by using the same procedure of synthesizing 1032, using aniline as a starting material.










N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


2,2-dimethylpropyl)-N2-phenyloxalamide, 1035
















MS (M + H)+ Calcd.
711.2


MS (M + H)+ Observ.
711.3


Retention Time
2.45 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









Synthesis of Compound 1036, N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-2,2-dimethylpropyl)-N2-(1-(pyrrolidin-1-ylmethyl)cyclopropyl)methyl)oxalamide




1036 was prepared by using the same procedure of synthesizing 1032, using (1-(pyrrolidin-1-ylmethyl)cyclopropyl)methanamine as a starting material.










N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-


2,2-dimethylpropyl)-N2-((1-(pyrrolidin-


1-ylmethyl)cyclopropyl)methyl)oxalamide, 1036
















MS (M + H)+ Calcd.
772.3


MS (M + H)+ Observ. 
772.4


Retention Time
2.11 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









The following selected examples were prepared by using the same procedure of synthesizing 1032, using the corresponding amines rather than cyclopropylmethanamine hydrochloride as starting materials:










LC Condition A
















Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u


























MS
MS
HPLC



Compd.

(M + H)+
(M + H)+
Rt



#
Structure
Calcd.
Obs.
(min)
Method







1479




729.2
729.5
2.07
A


 


1536




725.3
725.3
2.12
A


 


3002




665.2
665.5
1.78
A


 


3006




741.2
741.6
2.11
A


 


3007




741.2
741.5
2.03
A


 


3009




759.2
759.5
2.07
A


 


3010




732.2
732.5
1.91
A


 


3012




755.3
755.5
1.99
A


 


3013




769.3
769.6
2.04
A


 


3017




801.3
801.5
2.19
A


 


3032




739.2
739.5
1.54
A


 


3033




701.2
701.5
1.73
A


 


3036




715.2
715.5
1.81
A


 


3037




796.3
796.5
1.91
A


 


3038




762.3
762.5
1.94
A


 


3040




702.2
702.4
1.89
A


 


3041




753.3
753.5
1.73
A


 


3043




767.3
767.5
1.64
A


 


3071




763.2
763.2
2.25
A


 


3074




651.2
651.4
1.76
A


 


3141




815.3
815.4
2.15
A


 


3143




813.3
813.4
2.27
A


 


3146




777.2
777.3
2.01
A


 


3176




715.2
715.4
1.80
A


 


3178




715.2
715.4
1.77
A


 


3239




736.2
736.3
1.98
A


 


3241




712.2
712.3
1.83
A


 


3245




712.2
712.3
1.86
A


 


3248




712.2
712.3
1.97
A


 


3250




797.2
797.3
2.25
A


 


3253




702.2
702.3
1.52
A


 


3256




778.3
778.4
2.14
A


 


3262




765.2
765.3
2.10
A


 


3337




750.3
750.3
2.00
A


 


3350




772.2
772.4
2.15
A


 


3352




752.2
752.4
1.80
A


 


3355




765.2
765.4
2.04
A


 


3359




767.2
767.3
1.96
A


 


3361




797.3
797.4
1.75
A


 


3375




754.2
754.4
1.88
A


 


3376




768.2
768.4
2.16
A


 


3378




759.2
759.4
2.31
A


 


3400




830.3
830.4
2.05
A


 


3404




829.3
829.4
2.06
A


 


3452




763.2
763.4
2.17
A


 


3454




750.3
750.4
2.09
A


 


3805




782.2
782.2
2.23
A


 


3808




769.2
769.3
1.89
A


 


2824




785.2
786.2
2.10
A


 


3826




782.2
782.2
2.13
A


 


3836




743.3
743.3
1.99
A









Preparation of 1465, N1-(2-((4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)methyl)cyclopentyl)-N2-(4-fluorophenyl)oxalamide
iPr2NEt (0.2 mL) and 4-fluoroaniline (39.2 mg) added into the solution of ethyl 2-((2-((4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)methyl)cyclopentyl)amino)-2-oxoacetate (40 mg) in EtOH (2 mL) and the reaction was stirred at 155° C. for 3 days. The product was isolated by preparative HPLC system.














Compound 1465
















MS (M + H)+ Calcd.
741.2


MS (M + H)+ Observ. 
741.5


Retention Time
2.07 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Preparation of 1492, N1-((1-((4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)methyl)cyclopropyl)methyl)-N2-(4-fluorophenyl)oxalamide
To a solution of 2-(((1-((4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)methyl)cyclopropyl)methyl)amino)-2-oxoacetic acid (17 mg) and TBTU (10.33 mg) in DMF (2 mL) was added 4-fluoroaniline (3.58 mg). The mixture was stirred at room temperature for 16 hours, before being purified by preparative HPLC.














Compound 1492
















MS (M + H)+ Calcd. 
727.3


MS (M + H)+ Observ.
727.2


Retention Time
2.09 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 50


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









Preparation of 3388, N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamido)-2,2-dimethylpropyl)oxalamide




To a solution of 4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzoic acid (10 mg) and TBTU (7.74 mg) in DMF (1 mL) was added N1-(3-amino-2,2-dimethylpropyl)oxalamide (4.18 mg) and DIPEA (0.018 mL). The mixture was stirred at room temperature for 2 hours. The product was isolated by using preparative HPLC system.










Compound 3388
















MS (M + H)+ Calcd. 
653.2


MS (M + H)+ Observ.
653.0


Retention Time
2.11 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
 30


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um









The following selected examples were prepared by using the same synthetic route of synthesizing 3388, using the corresponding substituted alkyl 4-aminobenzoates rather than methyl 4-amino-2-fluorobenzoate or substituted benzamines rather than 1-(4-chlorophenyl)cyclopropanamine as starting materials:










LC Condition A


















Solvent A
90% Water-10% Methanol-0.1% TFA



Solvent B
10% Water-90% Methanol-0.1% TFA



Start % B
 30



Final % B
100



Gradient Time 
2 min



Flow Rate
1 mL/min



Wavelength
220



Solvent Pair 
Water-Methanol-TFA



Column
PHENOMENEX-LUNA 2.0 × 30 mm 3 um




















LC Condition B
















Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair 
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 50 × 2, 3 u


























MS
MS
HPLC



Compd.

(M + H)+
(M + H)+
Rt



#
Structure
Calcd.
Obs.
(min)
Method







3386




653.2
653.0
2.08
A


 


3387




665.2
665.0
2.11
A


 


3389




651.2
651.0
2.16
A


 


3477




669.2
669.3
2.16
A


 


3478




651.2
651.3
2.07
A


 


3511




711.2
711.4
2.08
A


 


3519




717.2
717.4
1.88
A


 


3812




671.2
671.2
3.25
B


 


3243




619.2
619.1
2.12
A


 


3244




601.2
601.2
2.06
A


 


3247




619.2
619.1
2.10
A


 


3265




635.2
635.1
2.11
A


 


3333




665.2
665.1
1.94
A


 


3351




699.2
699.2
2.01
A


 


4001




635.2
635.1
2.02
A









The following selected examples were prepared by using the same synthetic route of synthesizing 1032, using In-1006 rather than In-1005 as one of the starting materials, alone with the corresponding amines or their salts instead of cyclopropylmethanamine hydrochloride:










LC Condition A
















Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair 
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u


























MS (M + H)+
MS (M + H)+
HPLC Rt



Compd. #
Structure
Calcd.
Obs.
(min)
Method







3512




763.2
763.2
2.16
A


 


3513




781.2
781.1
2.19
A


 


3514




761.2
761.3
2.05
A


 


3515




755.2
755.3
2.54
A


 


3516




754.2
754.3
2.13
A


 


3553




797.2
797.3
2.23
A


 


3554




743.2
743.3
2.18
A


 


3555




765.2
765.2
2.23
A


 


3556




805.3
805.3
2.28
A


 


3557




759.2
759.3
2.06
A


 


3559




813.2
813.2
2.24
A


 


3560




789.3
789.3
2.12
A


 


3561




891.3
891.3
2.35
A


 


3562




795.2
795.3
2.08
A


 


3563




835.3
835.2
2.21
A


 


3564




831.3
831.3
2.31
A


 


3565




784.2
784.3
1.97
A


 


3566




786.2
786.2
2.00
A


 


3567




733.2
733.2
1.87
A


 


3568




786.2
786.2
2.09
A


 


3664




777.2
777.1
2.26
A


 


3676




802.2
802.3
1.87
A


 


3677




846.2
846.2
2.13
A


 


3678




865.2
865.1
2.10
A


 


3680




850.2
850.1
2.16
A


 


3681




854.2
854.1
2.29
A


 


3814




816.2
816.2
2.08
A









General Procedure for Parallel Synthesis of Oxalamides
Method 1 and Method 2
Method 1:
A 0.08 M stock solution of the (S)-methyl 2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-4-(2-ethoxy-2-oxoacetamido)butanoate in DMF was prepared and DIPEA (5.0 eq) was charged in. To each of the amines (15 eq) weighed into 16×100 mm threaded vials was added 1000 μLs of the (S)-methyl 2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-3-(2-ethoxy-2-oxoacetamido)propanoate/DIPEA solution. The reactions were agitated at 350 rpm on an Innova platform shaker at room temperature for 48 hours. Mixtures were transferred to a 96 well filter plate collecting into a 96 well deep-well plate. Each reaction vial was rinsed with 500 μLs of DMF and rinses were transferred to the appropriate wells of the filter plate. The desired products were isolated by preparative HPLC.
Method 2:
A 0.08 M stock solution of the (S)-methyl 2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-4-(2-ethoxy-2-oxoacetamido)butanoate in NMP was prepared and DIPEA (5.0 eq) was charged in. To each of the amines (15 eq) weighed into 16×100 mm threaded vials was added 1000 μLs of the (S)-methyl 2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-3-(2-ethoxy-2-oxoacetamido)propanoate/DIPEA solution. The reactions were agitated at 350 rpm on an Innova platform shaker at room temperature for 48 hours. Samples were blown down using a Zymark tabletop dryer at 30° C. for 3 hours. A 1.8M solution of potassium carbonate in water was prepared. To each of the reaction vials was added 0.5 mL of acetone followed by 0.5 mls of the potassium carbonate solution. Vials were capped and agitated at 350 rpm on an Innova platform shaker at room temperature for 72 hours. Samples were blown down using a Zymark tabletop dryer at 30° C. for 3 hours before redissolving in 1 mL DMF each. Mixtures were transferred to a 96 well filter plate collecting into a 96 well deep-well plate. Reaction vials were rinsed with 500 μLs of DMF each and rinses were transferred to the appropriate wells of the filter plate. The desired products were isolated by preparative HPLC.
Analytical Methods
A:
Column. Supelco Ascentis Express C18, 4.6×50 mm, 2.7-μm particles; Mobile
Phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 40° C.;
Gradient: 0-100% B over 8 minutes, then a 1-minute hold at 100% B, then 100-0% B over 1 minute; Flow: 1 mL/min.
B:
Column: Waters Sunfire C18, 4.6×100 mm, 3.5-μm particles; Mobile Phase A: water with 0.1% TFA; Mobile Phase B: acetonitrile:water with 0.1% TFA;
Temperature: 40° C.; Gradient: 10-95% B over 6 minutes, then a 2-minute hold at 95% B, then 95-10% B over 0.50 minutes, then a 1.5 minute hold at 10% B; Flow: 1.2 mL/min.
C:
Column. Waters BEH C18, 2.1×50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes, then a 0.5-minute hold at 100% B;
Flow: 1 mL/min.
D:
Column. Waters BEH C18, 2.1×50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient: 0-100% B over 4 minutes, then a 1-minute hold at 100% B; Flow: 0.83 mL/min.
E:
Column. Waters BEH C18, 2.1×50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient: 0-100% B over 4 minutes, then a 1-minute hold at 100% B; Flow: 1 mL/min.
F:
Column. Waters BEH C18, 2.0×50 mm, 1.7-μm particles; Mobile Phase A: 5:95 methanol:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 methanol:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes, then a 0.5-minute hold at 100% B;
Flow: 0.5 mL/min.
General Procedure for Parallel Synthesis of Oxalamides
Method 3




Step 1: To a solution of (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-(2-ethoxy-2-oxoacetamido)butanoate (1.0 g) in methanol (10 mL) was added Et3N (10.0 mL) at room temperature. The reaction mixture was heated to 55° C. for 24 hours. The solvent was concentrated in vacuo. The reaction mixture was diluted with water and acidified with 1.5; N HCl. The aqueous layer was extracted with ethylacetate (3×100 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo to afford desired crude acid as white solid (850 mg).
Step 2: To a solution of (S)-2-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-4-methoxy-4-oxobutylamino)-2-oxoacetic acid (20 mg) in DMF (1.0 mL) were added HCTU (17.91 mg) and DIPEA (16 μL) at rt and stirred for 15 minutes. Amine (1.2 eq.) was added to above solution and stirred at room temperature for 2 hour. Initial LCMS analysis showed formation of desired product. The solvent was evaporated and crude mixture was purified using preparative HPLC to afford desired product.
Step 3: To a solution of crude amide from step 2 in MeOH:H2O (5:1, 1.0 mL) was added K2CO3 (8.3 mg) and stirred for 24 hours at room temperature. The crude mixture was purified using preparative HPLC to afford desired product.
General Procedure for Parallel Synthesis of Oxalamides
Method 4




Step 1: To a solution of (S)-methyl 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-(2-ethoxy-2-oxoacetamido)propanoate (1.5 g) in methanol (15 mL) was added Et3N (15.0 mL) at room temperature. The reaction mixture was heated to 55° C. for 24 hours. The solvent was concentrated in vacuo. The reaction mixture was diluted with water and acidified with 1.5N HCl. The aqueous layer was extracted with ethylacetate (3×100 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo to afford desired crude acid as white solid (1.35 g).
Step 2: To a solution of (S)-2-(2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-3-methoxy-3-oxopropylamino)-2-oxoacetic acid (20 mg) in DMF (1.0 mL) were added HCTU (17.91 mg) and DIPEA (16 μL) at room temperature and stirred for 15 minutes. Amine (1.2 eq.) was added to above solution and stirred at room temperature for 2 hours. Initial LCMS analysis showed formation of desired product. The solvent was evaporated and crude mixture was purified using preparative HPLC to afford desired product.
Step 3: To a solution of crude amide from step 2 in MeOH:H2O (5:1, 1.0 mL) was added K2CO3 (8.3 mg) and stirred for 24 hours at room temperature. The crude mixture was purified using preparative HPLC to afford desired product.
Preparation of Compound 2001, 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-N-(2,2-dimethyl-3-(2-oxo-2-(pyrrolidin-1-yl)acetamido)propyl)-3-fluorobenzamide




Step 1:
To a solution of 2,4,6-trichloro-1,3,5-triazine (15 g) in THF (300 mL) was added 2,2,2-trifluoroethanol (8.14 g) and Hunig'sBase (15.63 mL). The resulting mixture was stirred for 16 hours. After removal of most THF and precipitate through a plug washing with THF, the filtrate was concentrate to give a crude that will be used as it is.
Step 2:
To a solution of the product in Step 1 above (1.47 g, 5.91 mmol) in THF (10 mL) was added methyl 4-amino-3-fluorobenzoate (1 g, 5.91 mmol) and Hunig'sBase (3.10 mL, 17.74 mmol). The resulting mixture was stirred for 16 h. The precipitate was filtered and washed with THF and water and dried to give 1.5 g of methyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzoate as a solid.










methyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-


2-ylamino)-3-fluorobenzoate


















MS (M + H)+ Calcd.
381.7



MS (M + H)+ Observ. 
381.0



Retention Time
1.0 min







LC Condition










Solvent A
100% Water:0.05% TFA



Solvent B
100% ACN:0.05% TFA



Start % B
 2



Final % B
 98



Gradient Time
2.2 min



Flow Rate
0.8 mL/min



Wavelength
220



Solvent Pair
ACN:Water:TFA



Column
Acquity UPLC BEH C18 1.7 uM










Step 3:
To a solution of methyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzoate (0.93 g, 2.45 mmol) in THF (10 mL) was added 1-(4-chlorophenyl)cyclopropanamine (0.5 g, 2.45 mmol) and Hunig's Base (1.71 mL, 9.8 mmol). The resulting mixture was refluxed at 80° C. for 16 hs. The precipitate was filtrated through a plug washing with THF to give a crude product that was purified by Biotage eluting with 4/1 to 3/2 hexane/ethyl acetate to give 1.2 g of methyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoro ethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzoate as product.










methyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzoate


















MS (M + H)+ Calcd. 
512.9



MS (M + H)+ Observ.
512.1



Retention Time
1.10 min







LC Condition










Solvent A
100% Water:0.05% TFA



Solvent B
100% ACN:0.05% TFA



Start % B
 2



Final % B
 98



Gradient Time
2.2 min



Flow Rate
0.8 mL/min



Wavelength
220



Solvent Pair
ACN:Water:TFA



Column
Acquity UPLC BEH C18 1.7 uM










Step 4:
To a solution of methyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzoate (1.2 g, 2.34 mmol) in THF/water (1:1, 10 mL) was added LiOH (0.6 g, 25.0 mmol). The reaction mixture was refluxed at 80° C. for 7 hours. After cooling to room temperature, the reaction solution was acidified with 1N HCl. The product was extracted by EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated under vacuum. The crude product 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzoic acid was used directly in the next step.










4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-ylamino)-3-fluorobenzoic acid
















MS (M + H)+ Calcd. 
498.8


MS (M + H)+ Observ.
498.0


Retention Time
1.00 min







LC Condition








Solvent A
100% Water:0.05% TFA


Solvent B
100% ACN:0.05% TFA


Start % B
 2


Final % B
 98


Gradient Time
2.2 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair 
ACN:Water:TFA


Column
Acquity UPLC BEH C18 1.7 uM









Step 5:
To a solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzoic acid (200 mg, 0.42 mmol) in DCM (3 mL) was added tert-butyl 3-amino-2,2-dimethylpropylcarbamate (98 mg, 0.48 mmol), HATU (229 mg, 0.60 mol) and iPr2NEt (0.35 mL, 2.00 mmol). The mixture was stirred at r.t. for 16 hours before all the solvents were removed under vacuum. All solvents were removed under vacuum and the residue was purified by silica gel column (EtOAC/Hexanes=20% to 40%) to give tert-butyl 3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamido)-2,2-dimethylpropylcarbamate (270 mg, 100%) as a white solid.










tert-butyl 3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-


(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-


fluorobenzamido)-2,2-dimethylpropylcarbamate


















MS (M + H)+ Calcd. 
683.1



MS (M + H)+ Observ.
682.3



Retention Time
1.15 min







LC Condition










Solvent A
100% Water:0.05% TFA



Solvent B
100% ACN:0.05% TFA



Start % B
 2



Final % B
 98



Gradient Time
2.2 min



Flow Rate
0.8 mL/min



Wavelength
220



Solvent Pair
ACN:Water:TFA



Column
Acquity UPLC BEH C18 1.7 uM










Step 6:
tert-butyl 3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamido)-2,2-dimethylpropylcarbamate (270 mg, 0.40 mmol) was dissolved in 4 M HCl in dioxane (5 mL) solution. After being stirred at room temperature for 3 hours, the mixture was concentrated to give N-(3-amino-2,2-dimethylpropyl)-4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamide as crude product.










N-(3-amino-2,2-dimethylpropyl)-4-(4-(1-(4-chlorophenyl)


cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-ylamino)-3-fluorobenzamide


















MS (M + H)+ Calcd. 
583.0



MS (M + H)+ Observ.
582.2



Retention Time
0.92 min







LC Condition










Solvent A
100% Water:0.05% TFA



Solvent B
100% ACN:0.05% TFA



Start % B
 2



Final % B
 98



Gradient Time
2.2 min



Flow Rate
0.8 mL/min



Wavelength
220



Solvent Pair
ACN:Water:TFA



Column
Acquity UPLC BEH C18 1.7 uM










Step 7:
To a solution of N-(3-amino-2,2-dimethylpropyl)-4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamide (80 mg, 0.14 mmol) and 2-oxo-2-(pyrrolidin-1-yl)acetic acid (20 mg, 0.137 mmol) in DMF (2 mL) were added O-(benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate (66 mg, 0.21 mmol) and iPr2NEt (0.050 ml, 0.28 mmol). After stirring at rt for 16 hs, the mixture was purified by preparative HPLC to give 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-N-(2,2-dimethyl-3-(2-oxo-2-(pyrrolidin-1-yl)acetamido)propyl)-3-fluorobenzamide (14.4 mg, 14%) as product. 1H NMR (400 MHz, CHLOROFORM-d) 8 ppm 1.01 (6H, s), 1.42 (4H, m), 1.81-2.02 (4H, m), 3.14 (2H, m), 3.29 (2H, d, J=6.0 Hz), 3.60 (2H, t, J=6.9 Hz), 3.91-4.12 (2H, m), 4.70-4.80 (2H, m), 7.14-7.38 (3H, m), 7.48-7.59 (1H, m), 7.69-7.80 (1H, m), 7.94-8.06 (1H, m), 8.24-8.31 (1H, m).










4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-ylamino)-N-(2,2-dimethyl-3-(2-oxo-2-(pyrrolidin-l-


yl)acetamido)propyl)-3-fluorobenzamide
















MS (M + H)+ Calcd. 
708.1


MS (M + H)+ Observ.
707.3


Retention Time
1.07 min







LC Condition








Solvent A
100% Water:0.05% TFA


Solvent B
100% ACN:0.05% TFA


Start % B
2


Final % B
98


Gradient Time
2.2 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
ACN:Water:TFA


Column
Acquity UPLC BEH C18 1.7 uM









Preparation of Compound 2002, N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamido)-2,2-dimethylpropyl)-N2,N2-dimethyloxalamide




To a solution of N-(3-amino-2,2-dimethylpropyl)-4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamide (20 mg, 0.034 mmol) and 2-(dimethylamino)-2-oxoacetic acid (4.83 mg, 0.041 mmol) in DMF (1 mL) were added O-(benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate (16.5 mg, 0.052 mmol) and iPr2NEt (0.012 ml, 0.069 mmol). After stirring at rt for 16 hs, the mixture was purified by preparative HPLC to give N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamido)-2,2-dimethylpropyl)-N2,N2-dimethyloxalamide (13.8 mg, 56%) as product. 1H NMR (400 MHz, CHLOROFORM-d) 8 ppm 1.03 (6H, s), 1.31-1.53 (4H, m), 3.06 (3H, s), 3.10-3.21 (2H, m), 3.29 (2H, d, J=6.0 Hz), 3.39 (3H, s), 4.75 (2H, m), 7.14-7.29 (3H, m), 7.33-7.48 (1H, m), 7.70-7.81 (1H, m), 7.92-8.04 (1H, m), 8.07-8.15 (1H, m).










N1-(3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-ylamino)-3-fluorobenzamido)-2,


2-dimethylpropyl)-N2,N2-dimethyloxalamide
















MS (M + H)+ Calcd. 
682.1


MS (M + H)+ Observ.
681.3


Retention Time
1.02 min







LC Condition








Solvent A
100% Water:0.05% TFA


Solvent B
100% ACN:0.05% TFA


Start % B
 2


Final % B
 98


Gradient Time
2.2 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair 
ACN:Water:TFA









Displacement of CF3CH2O moiety by benzyl alcohol:




A solution of N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-2,2-dimethylpropyl)-N2-(p-tolyl)oxalamide (520 mg) in THF was prepared. NaH (19.86 mg) and 500 μL of the N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-2,2-dimethylpropyl)-N2-(p-tolyl)oxalamide solution was added into 16×48 mm each threaded vial containing an individual alcohol (5-10 eq.). Vials were capped and allowed to shake at room temperature for 3 days. Samples were blown down in the Zymark tabletop dryer at 35° C. for 2 hours and products were purified by preparative HPLC systems.
Method M=Column: Waters BEH C18, 2.0×50 mm, 1.7-μm particles; Mobile Phase A: 5:95 methanol:water with 10 mM ammonium acetate; Mobile
Phase B: 95:5 methanol:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient 0.5 min hold at 0% B, 0-100% B over 4 minutes, then a 0.5-minute hold at 100% B; Flow: 0.5 mL/min.
Method A=Column: Waters BEH C18, 2.0×50 mm, 1.7-μm particles; Mobile Phase A: 5:95
acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5
acetonitrile:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes, then a 0.5-minute hold at 100% B; Flow: 1 mL/min.

















MS
MS
HPLC



Compd.

(M + H)+
(M + H)+
Rt



#
Structure
Calcd.
Obs.
(min)
Method







3840




799.3
798.9
3.61
A


 


3841




789.4
789.8
4.86
M


 


3842




761.3
761.8
4.75
M


 


3843




733.3
733.7
4.60
M


 


3844




767.3
767.9
4.68
M


 


3845




751.3
751.7
4.58
M


 


3846




751.3
751.8
4.59
M


 


3847




751.3
751.9
4.60
M


 


3848




747.3
747.8
4.68
M


 


3849




767.3
767.7
4.68
M


 


3850




767.3
767.7
4.71
M


 


3851




801.3
800.8
3.76
A


 


3852




828.3
828.0
4.54
M


 


3853




762.3
762.8
4.43
M


 


3839




775.3
775.8
3.92
A


 


4002




786.3
786.8
3.75
A









Preparation of 3827, N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-((4-isopropylbenzyl)oxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamido)-2,2-dimethylpropyl)-N2-(p-tolyl)oxalamide




To a suspension of (4-isopropylphenyl)methanol (24.46 mg) and NaH (5.92 mg) in THF (10 mL) was added N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamido)-2,2-dimethylpropyl)-N2-(p-tolyl)oxalamide (11 mg). The mixture was heated to reflux for 3 hours. The reaction was quenched by 5% NaHCO3/H2O and extracted with EtOAc (2×10 mL). The organic layers were combined, washed with brine (10 mL), dried over MgSO4 and concentrated. The residue was purified by preparative HPLC system to give Compound 3827.










Compound 3827
















MS (M + H)+ Calcd.
793.3


MS (M + H)+ Observ. 
793.4


Retention Time
2.38 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Preparations of 3828, 3829, 3830 and 3835
Compounds 3828, 3829, 3830 and 3835 were prepared under the same condition for the synthesis of Compound 3827, using N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamido)-2,2-dimethylpropyl)-N2-(4-methoxyphenyl)oxalamide, N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamido)-2,2-dimethylpropyl)-N2-(4-(trifluoromethyl)phenyl)oxalamide, N1-([1, 1′-biphenyl]-2-yl)-N2-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-((4-isopropylbenzyl)oxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamido)-2,2-dimethylpropyl)oxalamide and N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-((4-isopropylbenzyl)oxy)-1,3,5-triazin-2-yl)amino)-2-fluorobenzamido)-2,2-dimethylpropyl)-N2-(2,4-difluorophenyl)oxalamide as starting materials.












3828

















MS (M + H)+ Calcd.
809.3


MS (M + H)+ Observ.
809.3


Retention Time
2.32 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u










3829

















MS (M + H)+ Calcd.
847.3


MS (M + H)+ Observ.
847.3


Retention Time
2.44 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u










3830

















MS (M + H)+ Calcd.
855.4


MS (M + H)+ Observ.
855.3


Retention Time
2.51 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u










3835

















MS (M + H)+ Calcd.
815.3


MS (M + H)+ Observ.
815.3


Retention Time
2.29 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3u









Preparations of 3834 and 4003
Step 1: iPr2NEt (4 mL) was added into a solution of 2-(chloromethyl)-2-methyloxirane (0.745 g) and (2,4-dimethoxyphenyl)methanamine (2.338 g) in EtOH (20 mL). The reaction was stirred at room temperature for 16 hours, then 115° C. for 72 hours before being quenched by water. After removal of solvents under vacuum, the residue was purified by silica gel chromatography or preparative HPLC.
















 








 


1,3-bis((2,4-dimethoxybenzyl)amino)-2-methylpropan-2-ol
















MS (M + H)+ Calcd.
405.2


MS (M + H)+ Observ. 
405.2


Retention Time
2.79 min







LC Condition








Solvent A
90% Water-10% Methanol-0.1% TFA


Solvent B
10% Water-90% Methanol-0.1% TFA


Start % B
0


Final % B
100


Gradient Time
4 min


Flow Rate
0.8 mL/min


Wavelength
220


Solvent Pair
Water-Methanol-TFA


Column
PHENOMENEX-LUNA 2.0 × 50 mm 3 um









Step 2: iPr2NEt (0.5 mL) and HATU (238 mg) were added into the solution of 4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzoic acid (300 mg) and 1,3-bis((2,4-dimethoxybenzyl)amino)-2-methylpropan-2-ol (253 mg) in THF (10 mL). The reaction was stirred at room temperature for 24 hours. The product was isolated by silica gel chromatography.
















 








 








 


4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-


triazin-2-yl)amino)-N-(2,4-dimethoxybenzyl)-N-(3-((2,4-


dimethoxybenzyl)amino)-2-hydroxy-2-methylpropyl)benzamide
















MS (M + H)+ Calcd.
866.3


MS (M + H)+ Observ.
866.3


Retention Time
2.01 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Step 3: iPr2NEt (0.5 mL), HATU (21.93 mg) were added into the solution of 4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-N-(2,4-dimethoxybenzyl)-N-(3-((2,4-dimethoxybenzyl)amino)-2-hydroxy-2-methylpropyl)benzamide (50 mg) in THF (10 mL). The reaction was stirred at room temperature for 24 hours, before being quenched by NaHCO3 aqueous solution (10 mL). The aqueous phase was extracted by EtOAc (3×10 mL). The combined organic layer was dried over MgSO4 and concentrated under vacuum to give a residue which was used as was.
















 








 


N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-


1,3,5-triazin-2-yl)amino)-N-(2,4-dimethoxybenzyl)benzamido)-2-hydroxy-2-


methylpropyl)-N1-(2,4-dimethoxybenzyl)oxalamide
















MS (M + H)+ Calcd.
937.3


MS (M + H)+ Observ.
937.3


Retention Time
1.93 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Step 4: TFA (0.2 mL) was added into a solution of N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)-N-(2,4-dimethoxybenzyl)benzamido)-2-hydroxy-2-methylpropyl)-N1-(2,4-dimethoxybenzyl)oxalamide (50 mg) in CH2Cl2 (2 mL). The reaction was stirred at room temperature for 24 hours, before all the solvents were removed under vacuum. The residue was purified by preparative HPLC system.
















 








 





















Compound 3834








MS (M + H)+ Calcd.
637.2


MS (M + H)+ Observ.
637.1


Retention Time
1.58 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u







Compound 4003








MS (M + H)+ Calcd.
619.2


MS (M + H)+ Observ.
619.1


Retention Time
1.72 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Preparation of 3838, N1-(3-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-2,2-bis(hydroxymethyl)propyl)oxalamide




Compound 3838 was prepared by using the same synthetic route of synthesizing 3834, using 2,2-bis(((2,4-dimethoxybenzyl)amino)methyl)propane-1,3-diol instead of 1,3-bis((2,4-dimethoxybenzyl)amino)-2-methylpropan-2-ol as one of the starting materials. And, 2,2-bis(((2,4-dimethoxybenzyl)amino)methyl)propane-1,3-diol was prepared by using the same procedure of synthesizing 1,3-bis((2,4-dimethoxybenzyl)amino)-2-methylpropan-2-ol by using 2,2-bis(bromomethyl)propane-1,3-diol rather than 2-(chloromethyl)-2-methyloxirane as one of the starting materials.










Compound 3838
















MS (M + H)+ Calcd.
667.2


MS (M + H)+ Observ. 
667.3


Retention Time
1.55 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
 0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Preparations of 4004, N2-((2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)cyclopentyl)methyl)-N2-(4-fluorophenyl)oxalamide and 4005, N1-((2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)cyclopentyl)methyl)-N2-(cyclopropylmethyl)oxalamide
Step 1: iPr2NEt (0.5 mL) was added into a solution of tert-butyl (2-(aminomethyl)cyclopentyl)carbamate (300 mg) and ethyl 2-chloro-2-oxoacetate (191 mg) in THF (10 mL). The reaction was stirred at room temperature for 16 hours before being quenched by water. The aqueous layer was extracted with EtOAc (3×20 mL). The combined organic phase was dried over MgSO4 and concentrated under vacuum to give the crude ethyl 2-(((2-((tert-butoxycarbonyl)amino)cyclopentyl)methyl)amino)-2-oxoacetate which was used as was.




Step 2: Ethyl 2-(((2-((tert-butoxycarbonyl)amino)cyclopentyl)methyl)amino)-2-oxoacetate (440 mg) in TFA (1 mL) and CH2Cl2 (10 mL) was stirred at room temperature for 16 hours. After removal of solvents, the crude ethyl 2-(((2-aminocyclopentyl)methyl)amino)-2-oxoacetate was used as was.
















 


ethyl 2-(((2-aminocyclopentyl)methyl)amino)-2-oxoacetate
















MS (M + H)+ Calcd.
215.1


MS (M + H)+ Observ.
215.2


Retention Time
0.65 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Step 3: iPr2NEt (0.5 mL) was added into a solution of 4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzoic acid (300 mg), ethyl 2-(((2-aminocyclopentyl)methyl)amino)-2-oxoacetate (205 mg) and TBTU (201 mg) in THF (10 mL). The reaction was stirred at room temperature for 16 hours before being quenched by water. The aqueous layer was extracted with EtOAc (3×20 mL). The combined organic phase was dried over MgSO4 and concentrated under vacuum to give the crude ethyl 2-(((2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)cyclopentyl)methyl)amino)-2-oxoacetate which was used as was.
















 








 








 


ethyl 2-(((2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-


yl)amino)benzamido)cyclopentyl)methyl)amino)-2-oxoacetate
















MS (M + H)+ Calcd.
676.2


MS (M + H)+ Observ.
676.4


Retention Time
1.82 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Step 4: A mixture of ethyl 2-(((2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)cyclopentyl)methyl)amino)-2-oxoacetate (30 mg) and 4-fluoroaniline (49.3 mg) in EtOH (2 mL) was stirred at room temperature for 16 hours, then at 155° C. for 16 hours. Then, the product 4004 was isolated by preparative HPLC system.
















 








 





















Compound 4004








MS (M + H)+ Calcd.
741.2


MS (M + H)+ Observ.
741.5


Retention Time
2.09 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Preparation of 4005, N2-((2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)cyclopentyl)methyl)-N2-(cyclopropylmethyl)oxalamide
A mixture of ethyl 2-(((2-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)cyclopentyl)methyl)amino)-2-oxoacetate (30 mg) and cyclopropylmethanamine (31.6 mg) in EtOH (2 mL) was stirred at room temperature for 16 hours. Then, the product 4005 was isolated by preparative HPLC system.
















 








 





















Compound 4005








MS (M + H)+ Calcd.
701.3


MS (M + H)+ Observ.
701.5


Retention Time
1.98 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium 



Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium 



Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Preparation of 3492, (R)-4-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)-2-(2-((3-fluorophenyl)amino)-2-oxoacetamido)butanoic acid
Step 1: N,N-Di-iso-propylethylamine (0.054 g) and 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate(V) (0.079 g) were added into a solution of 4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzoic acid (0.1 g) and (R)-4-amino-2-((tert-butoxycarbonyl)amino)butanoic acid (0.045 g) in DMF (1 mL). The reaction was stirred at room temperature for 16 hours and (R)-2-((tert-butoxycarbonyl)amino)-4-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)butanoic acid was isolated by using preparative HPLC system.
















 








 








 


(R)-2-((tert-butoxycarbonyl)amino)-4-(4-((4-((1-(4-


chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-


yl)amino)benzamido)butanoic acid
















MS (M + H)+ Calcd.
680.2


MS (M + H)+ Observ.
680.2


Retention Time
1.55 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Step 2: TFA (0.340 mL) was added into a solution of (R)-2-((tert-butoxycarbonyl)amino)-4-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)butanoic acid (0.2 g) in DCM (1 mL). The reaction was stirred at room temperature for 2 hours. After removal of solvents, the residue was purified by preparative HPLC system.
















 








 


(R)-2-amino-4-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-


trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)butanoic acid
















MS (M + H)+ Calcd.
580.2


MS (M + H)+ Observ.
580.2


Retention Time
1.52 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









Step 3: iPr2NEt (8.91 mg) and 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate(V) (0.013 g) were added into a solution of (R)-2-amino-4-(4-((4-((1-(4-chlorophenyl)cyclopropyl)amino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-yl)amino)benzamido)butanoic acid (0.020 g) and 2-((3-fluorophenyl)amino)-2-oxoacetic acid (6.32 mg) in DMF (1 mL). The reaction was stirred at room temperature for 1 hour and compound 3492 was isolated by using preparative HPLC system.
















 








 





















Compound 3492








MS (M + H)+ Calcd.
745.2


MS (M + H)+ Observ.
745.2


Retention Time
1.61 min







LC Condition








Solvent A
5% ACN:95% Water:10 mM Ammonium Actetate


Solvent B
95% ACN:5% Water:10 mM Ammonium Actetate


Start % B
0


Final % B
100


Gradient Time
2 min


Flow Rate
1 mL/min


Wavelength
220


Solvent Pair
ACN:Water:Ammonium Actetate


Column
Phenomenex Luna C18, 30 × 2, 3 u









It will be evident to one skilled in the art that the present disclosure is not limited to the foregoing illustrative examples, and that it can be embodied in other specific forms without departing from the essential attributes thereof. It is therefore desired that the examples be considered in all respects as illustrative and not restrictive, reference being made to the appended claims, rather than to the foregoing examples, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

## Claims

1. A compound of formula I



where:
R1 is alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl, alkylcarbonyl, or benzyl wherein the benzyl moiety is substituted with 0-3 substituents selected from cyano, halo, alkyl, haloalkyl, cycloalkyl, alkenyl, alkynyl, hydroxy, alkoxy, and haloalkoxy;
R2 is alkyl, cycloalkyl, (Ar2)alkyl, (Ar2)cycloalkyl, ((Ar2)cycloalkyl)alkyl, ((Ar2)alkyl)cycloalkyl, or (((Ar2)alkyl)cycloalkyl)alkyl;
R3 is hydrogen, alkyl, or cycloalkyl;
R4 is hydrogen, alkyl, or cycloalkyl;
R5 is




 where ring A is a 4 to 7 membered alkylene ring attached to the indicated nitrogen atom and bonded to L;
R5a is alkyl or benzyl wherein the benzyl is substituted with 0-3 substituents selected from the group consisting of halo, alkyl, haloalkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, and haloalkoxy;
R6 is hydrogen, alkyl, or cycloalkyl;
R7 is alkyl, cycloalkyl, benzocycloalkyl, bicycloalkyl, (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl, or bridged bicycloalkyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, benzocycloalkyl, bicycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, NR9CO2R11, N(R10)(R11), CO2R9,) CON(R10)(R11), NR9CON(R10)(R11), SO2N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, N(R13)COAr3, and N(R13)SO2Ar3;
or R7 is hydrogen, N-alkoxycarbonylpiperidinyl, piperidinonyl, or Ar4;
R8 is hydrogen, alkyl, or cycloalkyl wherein alkyl or cycloalkyl is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, fused bicycloalkyl, bridged bicycloalkyl, spirocycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, CO2R9, N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, and N(R13)SO2Ar3;
or R7 and R8 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl;
R9 is hydrogen, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, ((hydroxyalkyl)alkoxy)alkoxy, ((alkoxy)alkoxy)alkoxy, or Ar3;
R10 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, alkoxycarbonyl, or Ar6;
R11 is hydrogen, alkyl, cycloalkyl, or Ar6;
or R10 and R11 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl;
R12 is hydrogen, alkyl, cycloalkyl, or Ar6;
R13 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, alkoxycarbonyl, or Ar6, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, and haloalkoxy, N(R15)(R16), and alkylCO;
R14 is hydrogen, alkyl, cycloalkyl, or Ar6;
or R13 and R14 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl;
R15 is hydrogen, alkyl, cycloalkyl, alkylcarbonyl, or alkoxycarbonyl;
R16 is hydrogen, alkyl or cycloalkyl;
or R15 and R16 taken together with the nitrogen to which they are attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine 1,1-dioxide, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolinyl, or isoindolinyl, and is substituted with 0-2 substituents selected from alkyl, alkylcarbonyl, and alkoxycarbonyl;
L is alkylene, cycloalkylene, (cycloalkyl)alkyl, (alkyl)cycloalkyl, or alkyl(cycloalkyl)alkyl, and is substituted with 0-2 substituents selected from alkoxy, hydroxy, CO2R12 and CONR13R14;
Ar1 is phenyl substituted with 1 CON(R5)(R6) and with 0-3 substituents selected from cyano, halo, alkyl, haloalkyl, cycloalkyl, hydroxy, alkoxy, haloalkoxy, and Ar3;
Ar2 is phenyl substituted with 0-3 substituents selected from cyano, halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, and haloalkoxy;
Ar3 is phenyl, biphenyl, terphenyl, naphthalenyl, furanyl, benzofuranyl, fluorenyl, fluorenonyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzoisothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, indolinyl, chromenonyl, or dibenzofuranyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, Ar5, OAr5, NR13Ar5, N(R13)COAr5, N(R13)SO2Ar5, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), N(R13)(R14), and PhCONHSO2;
or Ar3 is phenyl substituted with 1 substituent selected from benzyl, phenoxy, pyridyloxy, pyrimidyloxy, tetrazolyloxy, thiazolyl, phenylpyrazolyl, methyloxadiazolyl, thiadiazolyl, triazolyl, methyltriazolyl, tetrazolyl, pyridinyl, dimethoxypyrimdinyl, indolyl, indolinyl, and isoindolinyl;
Ar4 is phenyl, indanyl, tetrahydronaphthyl, isochromanyl, benzodioxolyl, pyridinyl, pyrazolyl, imidazolyl, or triazolyl, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, haloalkoxy, N(R13)(R14), and alkylCO;
Ar5 is phenyl, naphthalenyl, furanyl, benzofuranyl, azabenzofuranyl, thiophenyl, benzothiophenyl, azabenzothiophenyl, pyrrolyl, indolyl, azaindolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindazolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, or indolinyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, (CO2R12)alkyl, (CO2R12)alkenyl, (CON(R13)(R14))alkyl, phenyl, hydroxy, alkoxy, OAr6, NR13Ar6, alkylthio, haloalkoxy, haloalkylthio, alkylcarbonyl, CO2R12, COR12, SO2R12, CON(R13)(R14), SO2N(R13)(R14), and N(R13)(R14);
Ar6 is phenyl, naphthalenyl, furanyl, benzofuranyl, azabenzofuranyl, thiophenyl, benzothiophenyl, azabenzothiophenyl, pyrrolyl, indolyl, azaindolyl, indanyl, pyridinyl, quinolinyl, azaquinolinyl, isoquinolinyl, azaisoquinolinyl, quinoxalinyl, azaquinoxalinyl, pyrimidinyl, quinazolinyl, azaquinazolinyl, pyrazolyl, indazolyl, azaindazolyl, oxazolyl, benzoxazolyl, azabenzoxazolyl, isoxazolyl, benzoisoxazolyl, azabenzoisoxazolyl, imidazolyl, benzoimidazolyl, azabenzoimidazolyl, thiazolyl, benzothiazolyl, azabenzothiazolyl, isothiazolyl, benzoisothiazolyl, azabenzothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzotriazolyl, azabenzotriazolyl, tetrazolyl, or indolinyl, and is substituted with 0-5 substituents selected from cyano, halo, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, phenyl, hydroxy, alkoxy, aryloxy, alkylthio, haloalkoxy, haloalkylthio, and alkylcarbonyl;
or a pharmaceutically acceptable salt thereof.

2. A compound of claim 1 where R1 is haloalkyl or benzyl wherein the benzyl moiety is substituted with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy, and haloalkoxy; R2 is (Ar2)alkyl or (Ar2)cycloalkyl; R3 is hydrogen; R4 is hydrogen; R7 is alkyl, cycloalkyl, (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl, or bridged bicycloalkyl, and is substituted with 0-4 substituents selected from the group consisting of halo, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy, benzyloxy, CO2R9, N(R10)(R11), tetrahydrofuranyl, tetrahydropyranyl, Ar3, OAr3, NR13Ar3, N(R13)COAr3, N(R13)COAr3, N(R13)SO2Ar3 and Ar4; L is




and Ar1 is phenyl para-substituted with 1 CON(R5)(R6), or a pharmaceutically acceptable salt thereof.

3. A compound of claim 2 where R1 is haloalkyl or benzyl wherein the benzyl moiety is substituted with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy, and haloalkoxy; R2 is (Ar2)alkyl or (Ar2)cycloalkyl; R3 is hydrogen; R4 is hydrogen; R7 is Ar3 or Ar4; L is




and Ar1 is phenyl para-substituted with 1 CON(R5)(R6), or a pharmaceutically acceptable salt thereof.

4. A compound of claim 1 where R1 is trifluoroethyl or a pharmaceutically acceptable salt thereof.

5. A compound of claim 1 where R1 is benzyl substituted with 0-5 halo, alkyl, alkenyl, haloalkyl, alkoxy or haloalkoxy substituents, or a pharmaceutically acceptable salt thereof.

6. A compound of claim 1 where R2 is (Ar2)alkyl or (Ar2)cycloalkyl, or a pharmaceutically acceptable salt thereof.

7. A compound of claim 1 where R3 is hydrogen and R4 is hydrogen, or a pharmaceutically acceptable salt thereof.

8. A compound of claim 1 where R7 is Ar3, or a pharmaceutically acceptable salt thereof.

9. A compound of claim 1 where R7 is Ar4, or a pharmaceutically acceptable salt thereof.

10. A compound of claim 1 where L is




or a pharmaceutically acceptable salt thereof.

11. A compound of claim 1 where Ar1 is phenyl para-substituted with 1 CON(R5)(R6), or a pharmaceutically acceptable salt thereof.

12. A compound of claim 1 selected from the group consisting of











































or a pharmaceutically acceptable salt thereof.

13. A composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

14. A method of treating hepatitis C infection comprising administering a therapeutically effective amount of a compound of claim 1 to a patient infected with the HCV virus and suitable for therapy as understood by practitioners in the field of hepatitis and HCV infection.