# Patent

## Title
Benzodiazepine derivatives, preparation thereof and use thereof

## Abstract

Compounds of Formula (I) wherein R1, R3, R4, R5 and X are as defined in the specification, as well as salts, enantiomers thereof and pharmaceutical compositions including the compounds are prepared. They are useful in therapy, in particular in the management of pain

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a US National Stage of International Application No. PCT/SE02/02306 that was filed on Dec. 11, 2002. The International Application claims priority under 35 U.S.C. § 119(a) to Swedish Application No. 0104250-6 filed Dec. 14, 2001.
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention is directed to compounds that are useful in treating or preventing pain, septic shock, pancreatitis, edema, rhinitis, asthma, colitis, arthritis, hepatorenal syndrome, cancer, bacterial and viral infections, ulcerative colitis, and Alzheimer's Disease. More particularly, the present invention is directed to benzodiazepine derivatives that useful in treating pain.
2. Discussion of Relevant Art
Two types of bradykinin receptor are known: The B1 receptor and the B2 receptor. A number of reports indicate an important role for the B2 receptor in the pathophysiology of pain. [e.g. Hall, J. M., Morton, I. K. M. The pharmacology and immunopharmacology of kinin receptors. In: Farmer S G (Ed). The kinin system. London: Academic Press, 1997; 9-44]. Hence, compounds that are B2 antagonists are useful in the relief of pain, including chronic pain and acute pain, e.g., chronic inflammatory pain, neuropathic pain, back pain, migraine, cancer pain, visceral pain, arthritis pain and post-operative pain.


DETAILED DESCRIPTION OF THE INVENTION
Thus, the problem underlying the present invention was to develop new compounds that are novel kinin B2 antagonists.
Accordingly, in one aspect, the present invention provides compounds that are useful in treating pain.
Definitions
Unless specified otherwise within this specification, the nomenclature used in this specification generally follows the examples and rules stated in Nomenclature of Organic Chemistry, Sections A, B, C, D, E, F, and H, Pergamon Press, Oxford, 1979, which is incorporated by references herein for its exemplary chemical structure names and rules on naming chemical structures. Optionally, a name of a compound may be generated using a chemical naming program: ACD/ChemSketch, Version 5.09/September 2001, Advanced Chemistry Development, Inc., Toronto, Canada.
The term “Cm-n” or “Cm-n group” used alone or as a prefix, refers to any group having m to n carbon atoms, and having 0 to n multivalent heteroatoms selected from O, S, N and P, wherein m and n are O or positive integers, and n>m. For example, “C1-6” would refer to a chemical group having 1 to 6 carbon atoms, and having 0 to 6 multivalent heteroatoms selected from O, S, N and P.
The term “hydrocarbon” used alone or as a suffix or prefix, refers to any structure comprising only carbon and hydrogen atoms up to 14 carbon atoms.
The term “hydrocarbon radical” or “hydrocarbyl” used alone or as a suffix or prefix, refers to any structure as a result of removing one or more hydrogens from a hydrocarbon.
The term “alkyl” used alone or as a suffix or prefix, refers to monovalent straight or branched chain hydrocarbon radicals comprising 1 to about 12 carbon atoms. Unless otherwise specified, “alkyl” general includes both saturated alkyl and unsaturated alkyl.
The term “alkylene” used alone or as suffix or prefix, refers to divalent straight or branched chain hydrocarbon radicals comprising 1 to about 12 carbon atoms, which serves to links two structures together.
The term “alkenyl” used alone or as suffix or prefix, refers to a monovalent straight or branched chain hydrocarbon radical having at least one carbon-carbon double bond and comprising at least 2 up to about 12 carbon atoms.
The term “alkynyl” used alone or as suffix or prefix, refers to a monovalent straight or branched chain hydrocarbon radical having at least one carbon-carbon triple bond and comprising at least 2 up to about 12 carbon atoms.
The term “cycloalkyl,” used alone or as suffix or prefix, refers to a monovalent ring-containing hydrocarbon radical comprising at least 3 up to about 12 carbon atoms.
The term “cycloalkenyl” used alone or as suffix or prefix, refers to a monovalent ring-containing hydrocarbon radical having at least one carbon-carbon double bond and comprising at least 3 up to about 12 carbon atoms.
The term “cycloalkenyl” used alone or as suffix or prefix, refers to a monovalent ring-containing hydrocarbon radical having at least one carbon-carbon triple bond and comprising about 7 up to about 12 carbon atoms.
The term “aryl” used alone or as suffix or prefix, refers to a monovalent hydrocarbon radical having one or more polyunsaturated carbon rings having aromatic character, (e.g., 4n+2 delocalized electrons) and comprising 5 up to about 14 carbon atoms.
The term “arylene” used alone or as suffix or prefix, refers to a divalent hydrocarbon radical having one or more polyunsaturated carbon rings having aromatic character, (e.g., 4n+2 delocalized electrons) and comprising 5 up to about 14 carbon atoms, which serves to links two structures together.
The term “heterocycle” used alone or as a suffix or prefix, refers to a ring-containing structure or molecule having one or more multivalent heteroatoms, independently selected from N, O, P and S, as a part of the ring structure and including at least 3 and up to about 20 atoms in the ring(s). Heterocycle may be saturated or unsaturated, containing one or more double bonds, and heterocycle may contain more than one ring. When a heterocycle contains more than one ring, the rings may be fused or unfused. Fused rings generally refer to at least two rings share two atoms therebetween. Heterocycle may have aromatic character or may not have aromatic character.
The term “heteroalkyl” used alone or as a suffix or prefix, refers to a radical formed as a result of replacing one or more carbon atom of an alkyl with one or more heteroatoms selected from N, O, P and S.
The term “heteroaromatic” used alone or as a suffix or prefix, refers to a ring-containing structure or molecule having one or more multivalent heteroatoms, independently selected from N, O, P and S, as a part of the ring structure and including at least 3 and up to about 20 atoms in the ring(s), wherein the ring-containing structure or molecule has an aromatic character (e.g., 4n+2 delocalized electrons).
The term “heterocyclic group,” “heterocyclic moiety,” “heterocyclic,” or “heterocyclo” used alone or as a suffix or prefix, refers to a radical derived from a heterocycle by removing one or more hydrogens therefrom.
The term “heterocyclyl” used alone or as a suffix or prefix, refers a monovalent radical derived from a heterocycle by removing one hydrogen therefrom.
The term “heterocyclylene” used alone or as a suffix or prefix, refers to a divalent radical derived from a heterocycle by removing two hydrogens therefrom, which serves to links two structures together.
The term “heteroaryl” used alone or as a suffix or prefix, refers to a heterocyclyl having aromatic character.
The term “heterocylcoalkyl” used alone or as a suffix or prefix, refers to a heterocyclyl that does not have aromatic character.
The term “heteroarylene” used alone or as a suffix or prefix, refers to a heterocyclylene having aromatic character.
The term “heterocycloalkylene” used alone or as a suffix or prefix, refers to a heterocyclylene that does not have aromatic character.
The term “six-membered” used as prefix refers to a group having a ring that contains six ring atoms.
The term “five-membered” used as prefix refers to a group having a ring that contains five ring atoms.
A five-membered ring heteroaryl is a heteroaryl with a ring having five ring atoms wherein 1, 2 or 3 ring atoms are independently selected from N, O and S.
Exemplary five-membered ring heteroaryls are thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl, 1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl, 1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl.
A six-membered ring heteroaryl is a heteroaryl with a ring having six ring atoms wherein 1, 2 or 3 ring atoms are independently selected from N, O and S.
Exemplary six-membered ring heteroaryls are pyridyl, pyrazinyl, pyrimidinyl, triazinyl and pyridazinyl.
The term “substituted” used as a prefix refers to a structure, molecule or group, wherein one or more hydrogens are replaced with one or more C1-12hydrocarbon groups, or one or more chemical groups containing one or more heteroatoms selected from N, O, S, F, Cl, Br, I, and P. Exemplary chemical groups containing one or more heteroatoms include heterocyclyl, —NO2, —OR, —Cl, —Br, —I, —F, —CF3, —C(═O)R, —C(═O)OH, —NH2, —SH, —NHR, —NR2, —SR, —SO3H, —SO2R, —S(═O)R, —CN, —OH, —C(═O)OR, —C(═O)NR2, NRC(═O)R, oxo (═O), imino (═NR), thio (═S), and oximino (═N—OR), wherein each “R” is a C1-12hydrocarbyl. For example, substituted phenyl may refer to nitrophenyl, pyridylphenyl, methoxyphenyl, chlorophenyl, aminophenyl, etc., wherein the nitro, pyridyl, methoxy, chloro, and amino groups may replace any suitable hydrogen on the phenyl ring.
The term “substituted” used as a suffix of a first structure, molecule or group, followed by one or more names of chemical groups refers to a second structure, molecule or group, which is a result of replacing one or more hydrogens of the first structure, molecule or group with the one or more named chemical groups. For example, a “phenyl substituted by nitro” refers to nitrophenyl.
The term “optionally substituted” refers to both groups, structures, or molecules that are substituted and those that are not substituted.
Heterocycle includes, for example, monocyclic heterocycles such as: aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, imidazolidine, pyrazolidine, pyrazoline, dioxolane, sulfolane 2,3-dihydrofuran, 2,5-dihydrofuran tetrahydrofuran, thiophane, piperidine, 1,2,3,6-tetrahydro-pyridine, piperazine, morpholine, thiomorpholine, pyran, thiopyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dihydropyridine, 1,4-dioxane, 1,3-dioxane, dioxane, homopiperidine, 2,3,4,7-tetrahydro-1H-azepine homopiperazine, 1,3-dioxepane, 4,7-dihydro-1,3-dioxepin, and hexamethylene oxide.
In addition, heterocycle includes aromatic heterocycles, for example, pyridine, pyrazine, pyrimidine, pyridazine, thiophene, furan, furazan, pyrrole, imidazole, thiazole, oxazole, pyrazole, isothiazole, isoxazole, 1,2,3-triazole, tetrazole, 1,2,3-thiadiazole, 1,2,3-oxadiazole, 1,2,4-triazole, 1,2,4-thiadiazole, 1,2,4-oxadiazole, 1,3,4-triazole, 1,3,4-thiadiazole, and 1,3,4-oxadiazole.
Additionally, heterocycle encompass polycyclic heterocycles, for example, indole, indoline, isoindoline, quinoline, tetrahydroquinoline, isoquinoline, tetrahydroisoquinoline, 1,4-benzodioxan, coumarin, dihydrocoumarin, benzofuran, 2,3-dihydrobenzofuran, isobenzofuran, chromene, chroman, isochroman, xanthene, phenoxathiin, thianthrene, indolizine, isoindole, indazole, purine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, phenanthridine, perimidine, phenanthroline, phenazine, phenothiazine, phenoxazine, 1,2-benzisoxazole, benzothiophene, benzoxazole, benzthiazole, benzimidazole, benztriazole, thioxanthine, carbazole, carboline, acridine, pyrolizidine, and quinolizidine.
In addition to the polycyclic heterocycles described above, heterocycle includes polycyclic heterocycles wherein the ring fusion between two or more rings includes more than one bond common to both rings and more than two atoms common to both rings. Examples of such bridged heterocycles include quinuclidine, diazabicyclo[2.2.1]heptane and 7-oxabicyclo[2.2.1]heptane.
Heterocyclyl includes, for example, monocyclic heterocyclyls, such as: aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, pyrazolidinyl, pyrazolinyl, dioxolanyl, sulfolanyl, 2,3-dihydrofuranyl, 2,5-dihydrofuranyl, tetrahydrofuranyl, thiophanyl, piperidinyl, 1,2,3,6-tetrahydro-pyridinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyranyl, thiopyranyl, 2,3-dihydropyranyl, tetrahydropyranyl, 1,4-dihydropyridinyl, 1,4-dioxanyl, 1,3-dioxanyl, dioxanyl, homopiperidinyl, 2,3,4,7-tetrahydro-1H-azepinyl, homopiperazinyl, 1,3-dioxepanyl, 4,7-dihydro-1,3-dioxepinyl, and hexamethylene oxidyl.
In addition, heterocyclyl includes aromatic heterocyclyls or heteroaryl, for example, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, furyl, furazanyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl, 1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl, 1,3,4-thiadiazolyl, and 1,3,4 oxadiazolyl.
Additionally, heterocyclyl encompasses polycyclic heterocyclyls (including both aromatic or non-aromatic), for example, indolyl, indolinyl, isoindolinyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, 1,4-benzodioxanyl, coumarinyl, dihydrocoumarinyl, benzofuranyl, 2,3-dihydrobenzofuranyl, isobenzofuranyl, chromenyl, chromanyl, isochromanyl, xanthenyl, phenoxathiinyl, thianthrenyl, indolizinyl, isoindolyl, indazolyl, purinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, phenanthridinyl, perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, 1,2-benzisoxazolyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benzimidazolyl, benztriazolyl, thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrolizidinyl, and quinolizidinyl.
In addition to the polycyclic heterocyclyls described above, heterocyclyl includes polycyclic heterocyclyls wherein the ring fusion between two or more rings includes more than one bond common to both rings and more than two atoms common to both rings. Examples of such bridged heterocycles include quinuclidinyl, diazabicyclo[2.2.1]heptyl; and 7-oxabicyclo[2.2.1]heptyl.
The term “alkoxy” used alone or as a suffix or prefix, refers to radicals of the general formula —O—R, wherein R is selected from a hydrocarbon radical. Exemplary alkoxy includes methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, isobutoxy, cyclopropylmethoxy, allyloxy, and propargyloxy.
The term “amine” or “amino” used alone or as a suffix or prefix, refers to radicals of the general formula —NRR′, wherein R and R′ are independently selected from hydrogen or a hydrocarbon radical.
“Acyl” used alone, as a prefix or suffix, means —C(═O)—R, wherein R is an optionally substituted hydrocarbyl, hydrogen, amino or alkoxy. Acyl groups include, for example, acetyl, propionyl, benzoyl, phenyl acetyl, carboethoxy, and dimethylcarbamoyl.
Halogen includes fluorine, chlorine, bromine and iodine.
“Halogenated,” used as a prefix of a group, means one or more hydrogens on the group is replaced with one or more halogens.
“RT” or “rt” means room temperature.
A first ring group being “fused” with a second ring group means the first ring and the second ring share at least two atoms therebetween.
“Link,” “linked,” or “linking,” unless otherwise specified, means covalently linked or bonded.
Description of Preferred Embodiments
In one aspect, the present invention provides a compound of formula (I), pharmaceutically acceptable salts thereof, diasteriomers thereof, enantiomers thereof, or mixtures thereof:




wherein

R1 is selected from optionally substituted acyl, optionally substituted alkyl-oxycarbonyl, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted aryl; optionally substituted heterocyclyl; optionally substituted aryl-C1-6alkyl, and optionally substituted heterocyclyl-C1-6alkyl; or a divalent C1-12 group that together with a second nitrogen of X to form a ring;
X is a divalent group including a first nitrogen atom and the second nitrogen atom, wherein a first group (e.g., the 2H,1,4benzodiazepin-2-one group of formula (I)) is linked to the first nitrogen atom and R1 is linked to the second nitrogen atom, and wherein the first and second nitrogen atoms are separated by either one carbon atom, or two carbon atoms wherein said two carbon atoms have a double bond therebetween;
R3 is optionally substituted aryl, optionally substituted C1-12alkyl, optionally substituted C3-12cycloalkyl, or optionally substituted heterocyclyl;
R4 is, at each position, independently —H, halogen, optionally substituted alkyl, optionally substituted heteroalkyl, nitro, cyano, hydroxy, —OR6, —SR6, —S(═O)R6, —S(═O)2R6, —C(═O)R6, —C(═S)R6, —NR7R6, —C(═O)NR7R6, —NR7C(═O)R6, —SO2NR7R6, —NR7SO2R6, or —C(═O)OR6; and
R5, R6 and R7 are independently —H, optionally substituted C1-6alkyl.
In another aspect, the compounds of the present invention are those of formula (I), pharmaceutically acceptable salts thereof, diasteriomers thereof, enantiomers thereof, or mixtures thereof, wherein
R1 is selected from optionally substituted acyl, optionally substituted alkyl-oxycarbonyl, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted aryl; optionally substituted heterocyclyl; optionally substituted aryl-C1-6alkyl, and optionally substituted heterocyclyl-C1-6alkyl; or a divalent C1-12 group that together with a divalent R2 of X forms a portion of a ring;
X is represented by (i), (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), (xvi), or (xvii) below:







wherein R2 is selected from —H, optionally substituted C1-12alkyl, optionally substituted C1-12heteroalkyl, optionally substituted aryl, optionally substituted heterocyclyl, and a divalent C0-6group together with a divalent R1 to form the portion of the ring, wherein said divalent C0-6 group optionally includes one or more heteroatoms;
R3 is optionally substituted aryl, optionally substituted C1-12alkyl, optionally substituted C3-12cycloalkyl, or optionally substituted heterocyclyl;
R4 is, at each position, independently, —H, halogen, optionally substituted alkyl, optionally substituted heteroalkyl, nitro, cyano, hydroxy, —OR6, —SR6, —S(═O)R6, —S(═O)2R6, —C(═O)R6, —C(═S)R6, —NR7R6, —C(═O)NR7R6, —NR7C(═O)R6, —SO2NR7R6, —NR7SO2R6, or —C(═O)OR6; and
R5, R6 and R7 are independently —H, optionally substituted C1-6alkyl.
More particularly, the compound of the present invention is a compound of formula (I), wherein
R1 is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, optionally substituted carbazolyl, or a first divalent group selected from optionally substituted C1-12alkylene and optionally substituted C1-12heteroalkylene; wherein said phenyl, naphthyl, isoquinolyl, acridinyl, coumarinyl, and carbazolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino, wherein said C1-12alkylene and C1-12heteroalkylene are optionally substituted by C1-6alkyl, aryl-C1-6alkyl, aryl or heterocyclyl;
X is selected from formulas (i), (ii), (iii), (vi) and (xvii) below:




R2 is —H, C1-3alkyl, or a second divalent group selected from a single bond, an optionally substituted alkylene and an optionally substituted heteroalkylene; wherein said second divalent group together with said first divalent group forms a portion of a ring;
R3 is optionally substituted aryl, optionally substituted heteroaryl or optionally substituted cycloalkyl;
R4 is halogen, or C1-3alkyl; and
R5 is C1-3alkyl.
Most particularly, the compound of the present invention is a compound of formula (I), wherein
—X—R1 of formula (I) is selected from formulas (a), (b), (c), (d), (e), (f) and (g) below:








R1 is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said phenyl, naphthyl and isoquinolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino;
R2 is —H, or C1-3alkyl;




is a nitrogen containing heterocyclyl, which may be optionally substituted by one or more —R8, and which includes a bond on the nitrogen that links to other group of formula (I). Exemplary nitrogen containing heterocyclyls include, but is not limited to, piperazinyl, morpholinyl, poperidyl, and pyrrolidinyl.

R8 is —H, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted C1-6alkyl, —OH, or C1-6alkoxy, wherein R8 is optionally fused with the ring of




R3 is optionally substituted cyclohexyl, optionally substituted phenyl, optionally substituted pyridyl, optionally substituted thienyl, or optionally substituted pyrimidinyl, wherein said cyclohexyl, phenyl, pyridyl, thienyl and pyrimidinyl are optionally substituted by halogen, methoxy, or C1-3alkyl;
R4 is halogen; and
R5is methyl.
Specific examples of compounds of the present invention that may be used in practicing the present invention are listed in Table 1, below.







TABLE 1







Spreadsheet of combinatorially prepared compounds with LCMS analysis


of the reaction product.











Compound

Target
Mass
Rtn Time


#
Structure
Mass
Found?
(MS)














1




474.13
Yes
3.92


 


2




471.15
Yes
2.9


 


3




449.11
Yes
2.93


 


4




466.13
No


 


5




483.15
Yes
2.85


 


6




517.17
Yes
3.97


 


7




627.16
No


 


8




525.2
Yes
3.47


 


9




477.14
Yes
3.11


 


10




503.15
No


 


11




543.19
Yes
4.24


 


12




468.08
Yes
3.55


 


13




531.19
No


 


14




455.15
Yes
3.11


 


15




463.12
Yes
3.05


 


16




449.11
Yes
2.94


 


17




478.12
Tentative


 


18




449.11
Yes
2.89


 


19




552.17
Yes
4.5


 


20




469.17
Yes
2.85


 


21




469.13
Yes
2.78


 


22




499.14
Yes
3.3


 


23




586.14
Yes
3.75


 


24




532.17
Yes
3.7


 


25




561.16
Yes
3.87


 


26




477.14
Yes
3.17


 


27




469.17
Yes
2.78


 


28




509.2
Yes
2.74


 


29




504.15
Yes
3.43


 


30




502.13
Yes
2.96


 


31




571.14
Yes


 


32




509.2
Yes
4.05


 


33




629.18
Yes


 


34




571.08
No


 


35




531.19
Yes
4.8


 


36




565.19
No


 


37




533.17
Yes
3.87


 


38




547.18
Yes
3.98


 


39




521.15
Yes
4.02


 


40




513.16
Yes
3.22


 


41




593.2
Yes


 


42




455.15
Yes
2.79


 


43




428.11
Yes
2.91


 


44




495.19
Yes
2.62


 


45




538.16
Yes
4.64


 


46




505.15
Yes
3.31


 


47




457.17
Yes
3.02


 


48




477.14
Yes
3.17


 


49




571.14
Yes
4.49


 


50




471.19
Yes
2.9


 


51




521.15
Yes
4.02


 


52




519.19
Yes
3.97


 


53




545.17
Yes
3.49


 


54




441.14
Yes
2.94


 


55




521.15
Yes
3.92


 


56




477.14
Yes
3.08


 


57




456.14
Yes
3.34


 


58




492.14
Yes
3.47


 


59




521.13
Yes
3.04


 


60




497.2
Yes
2.7


 


61




452.12
Yes
2.69


 


62




443.15
Yes
2.51


 


63




513.14
Yes
3.97


 


64




483.19
Tentative
1.63


 


65




455.15
Yes
2.8


 


66




455.15
Yes
2.77


 


67




469.13
Yes
2.69


 


68




515.15
Yes
3.59


 


69




515.15
Yes
3.59


 


70




469.13
Yes
2.73


 


71




456.14
Tentative


 


72




471.15
Tentative


 


73




442.12
No


 


74




519.15
Tentative


 


75




533.17
No


 


76




538.19
No


 


77




512.21
No


 


78




525.16
Yes
6.5


 


79




574.19
No


 


80




504.15
Tentative


 


81




505.15
No


 


82




495.19
No


 


83




571.08
No


 


84




531.19
Yes
4.18


 


85




469.17
Yes
3.28


 


86




616.13
No


 


87




455.15
Yes


 


88




503.15
Yes
5.38


 


89




483.15
No


 


90




622.15
Yes
6.44


 


91




571.14
No


 


92




620.1
Yes
4.37


 


93




531.19
No


 


94




531.19
No


 


95




428.11
Tentative


 


96




572.14
Tentative
6.45


 


97




518.13
No


 


98




661.12
Yes


 


99




613.23
Yes
6.59


 


100




532.17
No


 


101




558.16
No


 


102




598.19
No


 


103




519.15
No


 


104




470.15
Yes


 


105




456.14
Yes


 


106




463.12
Yes


 


107




463.12
Yes
5.94


 


108




469.17
Yes
2.7


 


109




485.17
No


 


110




498.2
No


 


111




513.16
Tentative


 


112




512.14
Yes
4.21


 


113




469.17
Yes
2.55


 


114




512.14
No


 


115




469.17
No


 


116




504.17
No


 


117




476.08
Tentative


 


118




490.12
No


 


119




457.17
Yes
2.68


 


120




499.22
No


 


121




455.15
Yes
2.63


 


122




497.2
No


 


123




493.09
Yes
3.34


 


124




469.17
No


 


125




467.15
No


 


126




443.15
No


 


127




469.17
No


 


128




505.17
No


 


129




569.22
No


 


130




555.21
Yes
3.99


 


131




555.21
Yes
4.03


 


132




529.19
Yes
3.74


 


133




527.18
No


 


134




541.19
No


 


135




541.19
Yes
3.84


 


136




527.18
Tentative


 


137




541.19
Yes
3.84


 


138




599.2
Yes
3.87


 


139




541.19
Yes
3.75


 


140




539.18
No


 


141




599.2
Yes
3.87


 


142




572.18
No


 


143




556.14
No


 


144




558.16
No


 


145




501.16
No


 


146




577.19
No


 


147




527.18
Yes
3.46


 


148




587.2
No


 


149




541.19
Yes
3.59


 


150




474.13
No


 


151




466.1
Yes
3.54


 


152




541.1
Yes
2.63, 2.81


 


153




553.17
Tentative
4.13


 


154




412.11
Tentative
2.21


 


155




471.15
Yes
2.9


 


156




453.14
Yes
3.37, 4.15


 


157




441.14
Yes
2.61


 


158




442.12
No


 


159




470.19
Yes
4.55, 4.66


 


160




426.13
Yes
3.27


 


161




504.17
Yes
4.52


 


162




536.16
Yes
3.46


 


163




476.14
Yes
3.97


 


164




562.16
Yes
3.1, 3.35


 


165




462.13
Yes
3.64


 


166




414.13
No


 


167




460.13
Yes
3.08


 


168




427.12
Yes
2.63


 


169




416.11
Yes
2.7


 


170




440.14
Yes
3.59


 


171




454.16
No


 


172




398.1
No


 


173




538.16
Yes
4.35


 


174




474.15
Yes
3.23


 


175




424.11
Yes
3.66


 


176




484.13
Yes
3.13


 


177




454.16
Yes
3.87


 


178




498.15
Yes
3.44


 


179




438.09
Yes
2.66, 3.16


 


180




480.12
Yes
3.68


 


181




492.14
Yes
3.44


 


182




494.19
Yes
3.68, 4.04,4.78


 


183




440.14
Yes
3.5


 


184




440.14
Yes
3.59


 


185




414.13
No


 


186




454.16
No


 


187




410.1
Tentative
2.81, 3.33


 


188




410.1
Yes
3.03, 6.5


 


189




492.14
Yes
3.59


 


190




494.19
Yes
4.73


 


191




548.14
Yes
3.62


 


192




498.13
Yes
4.06


 


193




456.14
Yes
3.21


 


194




492.1
Yes
3.4


 


195




476.14
Yes
3.88


 


196




501.14
Yes
3.22, 3.43


 


197




512.14
Yes
4.26


 


198




474.13
Yes
3.83


 


199




412.11
Tentative
3.12


 


200




498.15
Yes
3.39


 


201




458.12
Yes
2.96


 


202




532.09
Yes
4.16


 


203




538.14
Yes
3.24


 


204




440.07
No


 


205




516.17
Yes
4.45


 


206




608.2
Yes
4.22


 


207




443.15
Yes
2.57


 


208




517.17
Yes
3.73


 


209




474.15
Yes
3.39


 


210




466.16
Yes
4.22


 


211




466.16
Yes
3.99


 


212




508.13
Yes
3.19


 


213




524.14
Yes
4.17


 


214




538.16
Yes
4.39


 


215




492.14
Yes
3.64


 


216




463.12
Yes
2.56, 2.82


 


217




552.17
Yes
4.82


 


218




516.12
Yes
3.37


 


219




534.09
Yes
4.24


 


220




428.14
Yes
3.61


 


221




478.12
Yes
3.41


 


222




540.04
Yes
4.21


 


223




500.13
Yes
3.08


 


224




472.13
Tentative
3.13


 


225




502.16
No


 


226




506.19
Yes
4.8


 


227




438.13
Yes
3.52


 


228




490.16
Yes
4.19


 


229




440.14
Yes
3.52


 


230




522.15
Yes
3.32


 


231




478.12
Yes
3.49


 


232




396.08
No


 


233




458.12
Yes
3.12


 


234




474.13
No


 


235




482.07
Yes
3.29, 3.86


 


236




464.14
Yes
2.68, 4.77


 


237




442.12
Yes
3.02


 


238




398.1
No


 


239




502.18
Tentative
4.31


 


240




508.12
Yes
3.99


 


241




496.09
Yes
4.09


 


242




446.1
Yes
3.38


 


243




471.19
Yes
2.86


 


244




529.15
Yes
3.41


 


245




572.14
Yes
4.16


 


246




538.11
Yes
4


 


247




526.02
No


 


248




468.17
No


 


249




518.17
Yes
3.85


 


250




492.14
Yes
3.74


 


251




516.1
Yes
3.9


 


252




462.13
Yes
3.68


 


253




430.12
Yes
3.03


 


254




443.15
Yes
2.77


 


255




426.13
Yes
3.4


 


256




400.11
No


 


257




466.1
Yes
2.83, 3.54


 


258




416.11
Yes
2.86


 


259




448.11
Yes
3.45


 


260




600.06
Yes
2.43, 3.84


 


261




626.28
Yes
6.29


 


262




386.1
No


 


263




425.11
Yes
2.84


 


264




506.15
Yes
4.05


 


265




456.14
Yes
2.77


 


266




454.16
Yes
3.79


 


267




454.16
Yes
3.97


 


268




444.14
Yes
3.28


 


269




444.1
Yes
2.8


 


270




476.14
Yes
3.86


 


271




456.14
Yes
3.32


 


272




442.16
Yes
3.8


 


273




470.19
Yes
4.39


 


274




441.14
Yes
2.93


 


275




454.07
Yes
3.35


 


276




466.13
Yes
2.81


 


277




442.12
Yes
3.01


 


278




456.14
Tentative
3.13


 


279




508.13
Yes
3.48


 


280




430.12
Yes
2.95


 


281




482.07
Yes
3.89


 


282




428.14
Yes
3.53


 


283




412.11
Yes
3.19


 


284




454.16
Yes
3.84


 


285




478.12
Yes
3.03, 3.16,4.05


 


286




538.16
Yes
4.6


 


287




476.14
Yes
3.87


 


288




440.14
Yes
3.68


 


289




442.16
Yes
3.91


 


290




456.14
Yes
3.16


 


291




455.06
No


 


292




414.13
Tentative
3.12


 


293




518.15
Yes
3.51


 


294




466.1
Yes
3.5


 


295




470.15
No


 


296




494.13
Yes
3.99


 


297




440.14
Yes
3.61


 


298




398.1
No


 


299




452.11
No


 


300




452.11
No


 


301




442.12
Yes
3.02


 


302




452.11
Yes
2.78, 3.41


 


303




416.11
Yes
2.68


 


304




458.06
No


 


305




512.14
Yes
4.17


 


306




478.12
Yes
3.01


 


307




458.12
Yes
2.86


 


308




454.16
Yes
4.01


 


309




456.14
Yes
2.77


 


310




476.14
Yes
3.8


 


311




412.11
Yes
3.18, 6.2


 


312




476.07
Yes
2.75


 


313




492.06
Yes
3.77


 


314




484.09
Yes
3.45


 


315




476.14
Yes
3.9


 


316




492.14
Yes
3.77


 


317




454.16
Yes
3.91


 


318




517.17
No


 


319




586.14
Yes
4


 


320




490.12
Yes
3.41, 3.63


 


321




439.09
Yes
3.33


 


322




446.12
Yes
2.96


 


323




472.13
Yes
3.14


 


324




476.14
Yes
3.99


 


325




494.19
No


 


326




506.12
Yes
3.83


 


327




443.12
Yes
2.68


 


328




464.12
Yes
2.92


 


329




535.18
Yes
2.99


 


330




516.1
Yes
4.03


 


331




529.19
Yes
3.51


 


332




555.21
Yes
3.91, 4.15


 


333




517.1
Yes
3.44


 


334




531.19
Yes
3.6, 3.76


 


335




573.16
Yes
4.27


 


336




615.17
Yes
4.19


 


337




539.13
Yes
3.94


 


338




565.15
Yes
4.19, 4.36


 


339




581.14
Yes
3.9


 


340




535.18
Yes
3.43


 


341




488.12
No


 


342




527.09
Yes
2.73


 


343




469.17
Yes
2.58


 


344




530.05
Yes
4.59


 


345




480.12
Yes
3.73


 


346




480.12
Yes
3.06, 3.76


 


347




573.16
Yes
3.02


 


348




518.19
Yes
4.85


 


349




531.15
Yes
2.69, 3,3.25, 3.36


 


350




477.14
Yes
2.94


 


351




496.09
Yes
4.02


 


352




531.15
Yes
2.59, 3.2,3.31


 


353




559.22
Yes
3.37


 


354




456.14
Yes
3.01


 


355




442.12
Yes
2.97


 


356




470.15
Yes
3.15


 


357




496.09
Yes
4.11


 


358




471.19
Yes
2.6


 


359




515.15
Yes
3.49, 3.69


 


360




476.14
Yes
4.01


 


361




425.08
Yes
2.7


 


362




452.12
Yes
2.94


 


363




497.11
Yes
4.37


 


364




438.1
Yes
3.45


 


365




441.05
Yes
3.27


 


366




455.06
Yes
3.49


 


367




471.07
No


 


368




509.05
No


 


369




424.09
Yes
3.15


 


370




508.09
No


 


371




500.06
Yes
3.59


 


372




575.06
Yes
2.87


 


373




587.13
No


 


374




446.07
No


 


375




505.11
Yes
2.96


 


376




487.1
Yes
3.29, 4.02


 


377




475.1
Yes
2.68


 


378




476.08
No


 


379




504.15
Yes
4.64


 


380




460.09
Yes
3.29


 


381




538.14
Yes
4.51


 


382




570.13
Yes
3.49


 


383




510.1
Yes
3.96


 


384




596.12
Yes


 


385




496.09
Yes
3.66


 


386




448.09
No


 


387




494.09
Yes
3.12


 


388




461.08
Yes
2.71


 


389




450.07
Yes
2.77


 


390




474.1
Yes
3.6


 


391




488.12
No


 


392




432.06
No


 


393




572.12
Yes
4.36


 


394




508.11
Yes
3.24


 


395




458.07
Yes
3.55


 


396




518.09
Yes
3.16


 


397




488.12
Yes
3.88


 


398




532.11
Yes
3.44


 


399




472.05
Yes
3.21


 


400




514.08
Yes
3.72


 


401




526.1
Yes
3.46


 


402




528.15
No


 


403




474.1
Yes
3.51


 


404




474.1
Yes
3.59


 


405




448.09
No


 


406




488.12
No


 


407




444.06
Yes
2.84, 3.26


 


408




444.06
Yes
3.06


 


409




526.1
Yes
3.64


 


410




528.15
Yes
4.73


 


411




582.1
Yes
3.63


 


412




532.09
Yes
4.08


 


413




490.1
Yes
3.23


 


414




526.06
Yes
3.44


 


415




510.1
Yes
3.92


 


416




535.1
Yes
2.86, 3.28,3.48


 


417




546.1
Yes
4.25


 


418




508.09
Yes
3.81


 


419




446.07
Tentative
3.1


 


420




532.11
Yes
3.41


 


421




492.08
Yes
3.03


 


422




566.06
Yes
4.19


 


423




572.11
Yes
3.3


 


424




474.03
No


 


425




550.14
Yes
4.43


 


426




642.16
Yes
4.22


 


427




477.12
Yes
2.65


 


428




551.13
Yes
3.74


 


429




508.11
Yes
3.42


 


430




500.12
Yes
4.24


 


431




500.12
Yes
4


 


432




542.09
Yes
3.25


 


433




558.1
Yes
4.2


 


434




572.12
Yes
4.4


 


435




526.1
Yes
3.69


 


436




497.08
Yes
2.68


 


437




586.14
Yes
4.83


 


438




550.08
Yes
3.41


 


439




568.05
Yes
4.27


 


440




462.1
Yes
3.66


 


441




512.08
Yes
3.46


 


442




574
Yes
4.24


 


443




534.09
Yes
3.14


 


444




506.09
Yes
3.24


 


445




536.12
No


 


446




540.15
Yes
4.79


 


447




472.09
Yes
3.55


 


448




524.12
Yes
4.19


 


449




474.1
Yes
3.53


 


450




556.11
Yes
3.38


 


451




512.08
Yes
3.52


 


452




430.04
Yes
3.11, 6.48


 


453




492.08
Yes
3.16


 


454




508.09
Yes
3.9


 


455




516.03
Yes
3.9


 


456




498.1
Yes
4.65, 6.23


 


457




476.08
Yes
3.08


 


458




432.06
No


 


459




536.14
No


 


460




542.08
Yes
3.25, 4.03


 


461




530.05
Yes
4.12


 


462




480.06
Yes
2.81, 3.43


 


463




505.15
Yes
2.93


 


464




563.11
Yes
3.43


 


465




606.1
Yes
4.14


 


466




572.07
Yes
3.96


 


467




559.98
Yes
3.86


 


468




502.14
No


 


469




552.13
Yes
3.83


 


470




526.1
Yes
3.75


 


471




550.06
Yes
3.92


 


472




496.09
Yes
3.72


 


473




464.08
Yes
3.08


 


474




477.12
Yes
2.85


 


475




460.09
Yes
3.44


 


476




434.07
No


 


477




500.06
Yes
3.59


 


478




450.07
Yes
2.93


 


479




482.07
Yes
3.49


 


480




634.02
Yes
3.89


 


481




660.25
No


 


482




420.06
No


 


483




459.07
Yes
2.89


 


484




540.12
Yes
4.07


 


485




490.1
Yes
2.85


 


486




488.12
Yes
1.84, 3.8


 


487




488.12
Yes
3.93


 


488




478.1
Yes
3.32


 


489




478.06
Yes
2.87


 


490




510.1
Yes
3.87


 


491




490.1
Yes
3.3


 


492




476.12
Yes
3.81


 


493




504.15
Yes
4.39


 


494




475.1
Yes
2.99


 


495




488.03
Yes
3.37


 


496




500.1
Yes
2.88


 


497




476.08
Yes
3.08


 


498




490.1
No


 


499




542.09
Yes
3.53


 


500




464.08
Yes
3


 


501




516.03
Yes
3.92


 


502




462.1
Yes
3.58


 


503




446.07
Yes
3.24


 


504




488.12
Yes
3.86


 


505




512.08
Yes
3.16


 


506




572.12
Yes
4.56


 


507




510.1
Yes
3.89


 


508




474.1
Yes
3.7


 


509




476.12
Yes
3.94


 


510




490.1
Yes
3.21


 


511




489.03
No


 


512




448.09
Tentative
2.76


 


513




552.12
Yes
3.53


 


514




500.06
Yes
3.54


 


515




504.12
Yes
3.34


 


516




528.1
Yes
3.98


 


517




474.1
Yes
3.61


 


518




432.06
No


 


519




486.07
No


 


520




486.07
No


 


521




476.08
Yes
3.07


 


522




486.07
Yes
3.46


 


523




450.07
Yes
2.75


 


524




492.02
Tentative
6.5


 


525




546.1
Yes
4.2


 


526




512.08
Yes
3.08


 


527




492.08
Yes
2.93


 


528




488.12
Yes
4.04


 


529




490.1
Yes
2.85


 


530




510.1
Yes
3.84


 


531




446.07
Yes
3.3


 


532




510.04
Yes
2.82


 


533




526.02
Yes
3.76


 


534




518.05
Yes
3.47


 


535




510.1
Yes
3.91


 


536




526.1
Yes
3.8


 


537




488.12
Yes
3.93


 


538




551.13
No


 


539




620.1
Yes
4.03


 


540




524.08
Yes
3.57


 


541




473.05
Yes
2.79, 3.36


 


542




480.08
Yes
3.02


 


543




506.09
Yes
3.19


 


544




510.1
Yes
4.02


 


545




528.15
No


 


546




540.08
Yes
3.86


 


547




477.08
Yes
2.76


 


548




498.08
Yes
2.97


 


549




569.14
Yes
3.06


 


550




550.06
Yes
4.06


 


551




563.15
Yes
3.57


 


552




589.17
Yes
3.89, 4.18


 


553




551.06
Yes
3.49


 


554




565.15
Yes
3.68


 


555




607.12
Yes
4.24


 


556




649.13
Yes
4.2


 


557




573.09
Yes
3.92


 


558




599.11
Yes
4.3


 


559




615.1
Yes
3.93


 


560




569.14
Yes
3.46


 


561




522.08
Yes
3.32


 


562




561.05
Yes
2.79


 


563




503.13
Tentative
2.68


 


564




564.01
Yes
4.61


 


565




514.08
Yes
3.75


 


566




514.08
Yes
3.8


 


567




607.12
Yes
3.86


 


568




552.15
Yes
4.85


 


569




565.11
Yes
3.41


 


570




511.1
Yes
3


 


571




530.05
Yes
4.03


 


572




565.11
Yes
3.36


 


573




593.18
Yes
3.43


 


574




490.1
Yes
3.08


 


575




476.08
Yes
3.03


 


576




504.12
Yes
3.21


 


577




530.05
Yes
4.14


 


578




505.15
Yes
2.69


 


579




549.12
Yes
3.54, 3.74


 


580




510.1
Yes
4.03


 


581




472.06
Yes
2.87


 


582




483.19
Yes
2.57


 


583




477.1
Yes
2.38, 2.62


 


584




469.2
Yes
2.63, 5.03


 


585




401.11
Yes
2.45


 


586




477.1
Yes
2.67, 3.16


 


587




473.13
No


 


588




455.2
Yes
2.56, 3.84


 


589




429.14
Yes
2.48


 


590




427.12
Yes
2.49


 


591




441.14
Yes
2.5


 


592




441.1
Yes
2.48, 2.92


 


593




427.12
Yes
2.62


 


594




441.14
Yes
2.58


 


595




499.1
Yes
2.81, 3.84


 


596




427.12
Yes
2.53


 


597




441.14
Yes
2.59


 


598




487.14
No


 


599




439.12
Yes
2.56


 


600




499.14
Yes
2.81


 


601




441.14
Yes
2.53


 


602




497.2
No


 


603




491.15
Tentative
2.87


 


604




483.19
Tentative
2.63


 


605




415.1
Yes
2.49, 3.01


 


606




491.15
No


 


607




487.14
No


 


608




469.17
Yes
2.64


 


609




443.15
Yes


 


610




441.14
Yes
2.76


 


611




455.15
Yes
2.68


 


612




455.15
Yes
2.76


 


613




441.14
Yes
2.93


 


614




455.15
Yes
2.8


 


615




513.16
Yes
3.13


 


616




441.14
Tentative
2.9


 


617




455.15
Yes
2.9


 


618




501.16
No


 


619




453.14
Yes
2.73


 


620




513.16
Yes
3.13


 


621




455.15
No


 


622




525.23
Yes
3.15


 


623




519.19
Yes
2.72


 


624




511.22
Yes
3.19


 


625




443.15
Yes
2.64


 


626




519.19
Tentative


 


627




515.18
Tentative
3.73


 


628




497.2
Yes
2.66, 2.88


 


629




471.19
Yes
2.76


 


630




469.17
Yes
2.76


 


631




483.19
Yes
2.68


 


632




483.19
Yes
2.78


 


633




469.2
Yes
2.51, 3.27


 


634




483.2
Yes
2.54, 2.91


 


635




541.19
Yes
3.79


 


636




469.2
Yes
2.76, 3.1


 


637




483.2
Yes
2.5, 3.09


 


638




529.19
No


 


639




481.2
Yes
2.5, 2.97


 


640




541.2
Yes
3.79, 4.07


 


641




483.2
Tentative
2.4, 2.98


 


642




524.21
Yes
2.82


 


643




518.17
Yes
2.64


 


644




510.2
Tentative
2.9


 


645




442.13
Yes
2.48


 


646




518.17
Yes
2.68


 


647




514.16
No


 


648




496.18
Yes
2.6


 


649




470.17
Yes
2.52


 


650




468.2
Yes
2.24, 2.53


 


651




482.2
Yes
2.15, 2.52


 


652




482.17
Yes
2.65


 


653




468.15
Yes
2.52


 


654




482.17
Yes
2.55


 


655




540.17
Yes
2.66


 


656




468.15
Yes
2.52


 


657




482.17
Yes
2.52


 


658




528.17
No


 


659




480.15
Yes
2.51


 


660




540.17
Yes


 


661




482.17
Yes
2.58


 


662




492.16
Yes
3.63


 


663




468.12
Yes
2.99


 


664




537.15
Yes
3.64


 


665




430.15
Yes
2.78


 


666




468.06
Tentative
3.33


 


667




468.06
Yes
3.76


 


668




460.16
Yes
2.76


 


669




468.06
Yes


 


670




430.15
No


 


671




428.17
Yes
3.08


 


672




445.12
Yes
3.03


 


673




478.05
No


 


674




425.13
Yes
2.89


 


675




458.14
No


 


676




471.21
Yes
3.41


 


677




444.16
Yes
3.01


 


678




460.16
Yes
2.68


 


679




434.1
Tentative
3.32


 


680




418.13
Yes
3.01


 


681




468.12
Yes
3.44


 


682




434.1
Yes
3.21, 6.45


 


683




434.1
Yes
2.93, 6.48


 


684




444.13
Yes
2.73


 


685




493.19
Yes
3.31


 


686




430.15
Yes
2.79


 


687




506.18
Yes
3.7


 


688




536.05
Yes
4.46


 


689




526.1
Yes
2.53, 4.15


 


690




502.08
Yes
3.39


 


691




571.11
No


 


692




464.11
Yes
3.16


 


693




502.02
Yes
3.81


 


694




502.02
Yes


 


695




494.1
Yes
3.14, 3.45


 


696




502.02
No


 


697




464.11
Yes
3.34


 


698




462.13
Yes
3.49


 


699




479.08
No


 


700




512.01
Yes
3.36


 


701




459.09
Yes
3.27


 


702




492.1
Yes
3.37, 6.53


 


703




505.17
Yes
3.86


 


704




478.12
Yes
3.46


 


705




494.12
Yes
2.99


 


706




468.06
Yes
3.76


 


707




452.09
Yes
3.45


 


708




502.08
Yes
3.92


 


709




468.06
Yes
3.74


 


710




468.06
Yes
3.34


 


711




478.1
Yes
3.12, 3.49


 


712




527.2
Yes
3.76, 4.81


 


713




464.1
Yes
3.19, 3.53


 


714




540.1(blank)
Tentative
6.4


 


715




508.13
Yes
4.35, 6.57


 


716




498.21
Yes
4.18


 


717




474.17
Yes
3.4


 


718




543.19
Yes
2.52, 4.19


 


719




436.19
Yes
2.7, 3.18


 


720




474.1
Yes
3.76


 


721




474.1
Yes
4.21


 


722




466.2
Yes
3.2


 


723




474.1
Yes
3.96


 


724




436.19
Yes
3.4


 


725




434.21
Yes
3.57


 


726




451.17
Yes
3.45


 


727




484.09
Yes
3.37


 


728




431.18
Yes
3.23


 


729




464.19
Yes
3.34


 


730




477.26
Yes
3.98


 


731




450.21
Yes
3.48


 


732




466.2
Yes
3.04


 


733




440.14
Yes
3.72


 


734




424.17
Yes
3.41


 


735




474.17
Yes
3.87


 


736




440.14
Yes
3.7


 


737




440.14
Yes
3.35


 


738




450.17
Yes
3.09


 


739




499.24
Yes
3.87


 


740




436.19
Yes
3.18


 


741




512.22
No


 


742




449.11
No


 


743




498.07
Yes
3.74


 


744




581.11
No


 


745




482.07
Yes
4


 


746




436.09
Yes
2.98


 


747




492.1
Yes
3.1


 


748




518.07
Yes


 


749




485.11
Yes
3.02


 


750




485.11
Yes
3.63


 


751




499.12
Yes
3.42


 


752




488.14
Yes
3.86


 


753




469.05
Yes
3.4


 


754




516.1
No


 


755




459.09
Yes
3.27


 


756




536.11
Yes
3.51


 


757




490.16
Yes
4.28


 


758




612.14
Yes
2.89


 


759




448.11
Yes
3.44


 


760




546.22
Tentative
6.44


 


761




532.09
Yes
4.11


 


762




526.12
No


 


763




526.12
Yes
4.19


 


764




502.08
Yes
4.11


 


765




482.07
Yes
3.67


 


766




526.12
Yes
3.31


 


767




483.07
Yes
3.44


 


768




526.02
Yes
4.15


 


769




506.12
No


 


770




473.11
No


 


771




522.13
Yes
3.06


 


772




483.07
No


 


773




526.12
Yes
3.37


 


774




559.99
Yes
3.93


 


775




510.13
Yes
4.2


 


776




534.15
Yes
4.55


 


777




462.13
Yes


 


778




600.04
No


 


779




509.11
Yes
3.57


 


780




524.14
Yes
3.92


 


781




550.11
Yes
3.55


 


782




478.12
No


 


783




473.11
Tentative
2.93


 


784




549.14
Yes
3.87


 


785




550.11
Yes
3.57


 


786




514.12
Yes


 


787




434.1
Yes
3.21, 6.52


 


788




452.09
Yes
3.45


 


789




464.11
Yes
3.27, 3.62


 


790




512.01
Yes
3.82


 


791




524.13
Yes
3.11, 3.58


 


792




538.12
Yes
4.05


 


793




502.02
Yes
4.3


 


794




485.11
Yes
3.62


 


795




466.1
Yes
3.43


 


796




460.11
Yes
3.66


 


797




504.01
Yes
6.43


 


798




502.02
Yes
4.55


 


799




493.05
Yes
3.41


 


800




545.97
Yes
4.41


 


801




528.08
Yes
3.39


 


802




482.07
Yes
3.44, 3.71


 


803




553.11
Yes
3.65


 


804




493.05
Yes
3.9


 


805




524.12
Yes
4.35, 6.49


 


806




498.07
Yes
3.42


 


807




482.07
Yes
3.99


 


808




468.06
Yes
3.74


 


809




502.02
Yes
3.92


 


810




482.07
Yes
3.68


 


811




502.02
Yes
4.32


 


812




550.09
No


 


813




482.07
Yes
3.84


 


814




506.12
Yes
3.25


 


815




482.07
Yes
3.62


 


816




506.12
Yes
3.57, 4.02


 


817




572.08
Yes
4.36


 


818




551.06
Yes
3.92


 


819




516.03
Yes


 


820




463.12
Yes
3.69


 


821




449.11
Yes
3.39


 


822




435.09
Yes
3.14


 


823




477.14
Yes
4.18


 


824




609.06
Tentative
3.91


 


825




485.11
Yes
3.69


 


826




485.11
Yes
3.82


 


827




449.11
Yes
3.37


 


828




482.07
Yes
3.86


 


829




463.12
Yes
3.64


 


830




541.13
No


 


831




449.11
Yes
3.4


 


832




463.12
Yes
3.81


 


833




527.06
No


 


834




449.11
Yes
3.45


 


835




485.11
No


 


836




486.1
No


 


837




496.11
No


 


838




464.11
Yes
3.15


 


839




478.12
Yes
2.76, 3.53,3.93


 


840




513  
Yes
3.75


 


841




608.14
Yes
3.96


 


842




518.08
Yes
3.29


 


843




592.04
Yes
3.8


 


844




462.13
Yes
3.76, 4.37


 


845




498.13
Yes
3.99, 4.74


 


846




516.1
No


 


847




561.08
No


 


848




532.09
Tentative
3.92


 


849




484.09
Yes
3.56


 


850




532.09
No


 


851




522.1
No


 


852




503.01
Yes
3.44


 


853




550.06
No


 


854




493.055
Yes
3.3


 


855




570.07
No


 


856




524.12
Yes
4.3


 


857




646.1
Yes
2.96


 


858




482.07
Yes
3.47


 


859




580.18
Yes
5.17


 


860




566.06
Yes


 


861




560.08
No


 


862




560.08
Yes


 


863




536.05
Yes
4.12


 


864




516.03
Tentative
3.67


 


865




560.08
Yes
3.33


 


866




517.03
Yes
3.38, 3.96


 


867




559.98
Tentative
4.16


 


868




540.08
No


 


869




507.07
No


 


870




556.09
Yes
4.26


 


871




517.03
No


 


872




560.08
Yes
3.39


 


873




593.95
No


 


874




544.09
Yes
4.21


 


875




568.11
Yes
4.55


 


876




496.09
No


 


877




634  
No


 


878




543.07
Yes
3.59


 


879




558.1
Yes
3.94


 


880




584.07
Yes
3.57


 


881




512.08
No


 


882




507.07
Yes
3.01


 


883




583.1
Yes
3.93


 


884




584.07
Yes
3.5


 


885




548.08
Tentative
4.47


 


886




468.06
Yes
3.24


 


887




486.05
Yes
3.48


 


888




498.07
Yes
3.29


 


889




545.97
No


 


890




558.09
Yes
2.02, 3.17,3.48


 


891




572.08
Yes
4.07


 


892




535.98
Yes
4.31


 


893




519.07
Yes
3.61


 


894




500.06
Yes
3.46


 


895




494.07
Yes
3.69


 


896




537.97
No


 


897




535.98
No


 


898




527.01
Yes
3.44


 


899




579.93
Tentative
4.41


 


900




562.04
Yes
3.41


 


901




516.03
Tentative
3.71


 


902




587.07
Yes
3.68


 


903




527.01
Yes
3.9


 


904




558.08
Yes
4.37


 


905




532.03
Yes
3.46


 


906




516.03
Tentative
3.99


 


907




502.02
Yes
3.76


 


908




535.98
Yes
3.93


 


909




516.03
Tentative
3.68


 


910




535.98
Yes


 


911




584.05
No


 


912




516.03
Yes
3.84


 


913




540.08
Yes
3.29


 


914




516.03
Yes
3.64


 


915




540.08
Yes
3.58


 


916




606.05
Yes
4.4


 


917




585.02
Yes
3.92


 


918




550  
Yes
4.25


 


919




497.08
Yes
3.67


 


920




483.07
Yes
3.36


 


921




469.05
Yes
3.14


 


922




511.1
Yes
4.05


 


923




643.02
No


 


924




519.07
Yes
3.65


 


925




519.07
Yes
3.83


 


926




483.07
Yes
3.35


 


927




516.03
Yes
3.85


 


928




497.08
Yes
3.59


 


929




575.09
No


 


930




459.04
Yes
3.11


 


931




483.07
Yes
3.4


 


932




486.08
Yes
2.99


 


933




497.08
Yes
3.71


 


934




531.07
Yes
4.27


 


935




475.01
Yes
3.28


 


936




561.02
No


 


937




483.07
Yes
3.42


 


938




489.03
Yes
3.47


 


939




505.03
Yes


 


940




543.01
No


 


941




519.07
Yes
3.87


 


942




520.06
No


 


943




458.05
Yes
2.78


 


944




530.07
No


 


945




498.07
Yes
3.2


 


946




512.08
Yes
3.55


 


947




546.96
Yes
3.69


 


948




642.1
Yes
3.98


 


949




552.04
Yes
3.34


 


950




626.01
Yes
3.79


 


951




496.09
Yes
3.77, 4.2


 


952




532.09
Yes
4, 4.52


 


953




550.06
Yes
3.74


 


954




595.05
No


 


955




566.06
Yes
3.91


 


956




518.05
Yes
3.59


 


957




566.06
No


 


958




474.13
Yes
4


 


959




460.11
Yes
3.9


 


960




494.07
Yes
3.9


 


961




350.17
Tentative
2.46


 


962




466.2
Yes
3.55, 6.41


 


963




364.19
Tentative
2.68


 


964




442.24
Yes
3.55


 


965




412.15
Tentative
2.78


 


966




390.21
Tentative
2.86


 


967




420.25
Tentative
3.55


 


968




412.19
Tentative
2.88


 


969




462.21
Yes
3.21


 


970




378.21
Tentative
2.91


 


971




424.03
No


 


972




394.16
Tentative
2.4


 


973




422.2
Tentative
2.63


 


974




472.2
Yes
2.15, 3.38


 


975




474.21
Yes
3.23


 


976




384.1
Tentative
2.75, 3.26


 


977




500.2
Yes
3.64, 4.02


 


978




398.2
Tentative
3.02, 3.66


 


979




476.2
Yes
4.06


 


980




446.11
Yes
3.11


 


981




424.2
Tentative
3.23, 3.88


 


982




454.2
Yes
4.06, 4.8


 


983




446.2
Yes
3.22, 3.85


 


984




496.2
Yes
3.6, 3.71


 


985




412.2
Tentative
3.28, 3.93


 


986




457.99
No


 


987




428.1
Yes
2.68, 3.08


 


988




456.2
Yes
2.89, 3.34,3.73


 


989




506.15
Yes
3.84


 


990




508.17
Yes
3.65


 


991




356.22
Tentative
2.81


 


992




472.28
Yes
4.11, 6.46


 


993




370.24
Tentative
3.05


 


994




448.28
Yes
4.07


 


995




418.2
Tentative
3.22


 


996




396.25
Tentative
3.29


 


997




426.3
Yes
4.09


 


998




418.24
Yes
3.29


 


999




468.25
Yes
3.67


 


1000




384.25
Tentative
3.35


 


1001




430.07
No


 


1002




400.21
Yes
2.72


 


1003




428.24
Tentative
3.08


 


1004




478.24
Yes
3.85


 


1005




480.25
Yes
2.7, 3.73


 


1006




384.16
Tentative
2.74


 


1007




474.19
Yes
2.7


 


1008




468.25
Yes
3.73


 


1009




402.15
Yes
2.81


 


1010




502.01
Yes
3.06, 3.9


 


1011




456.2
Yes
2.15, 3.05


 


1012




452.15
Yes
3.32


 


1013




452.15
No


 


1014




452.15
Yes
2.96


 


1015




398.17
Tentative
2.8


 


1016




444.18
Yes
2.76


 


1017




434.17
Tentative
3.03


 


1018




402.15
Tentative
2.9


 


1019




412.19
Yes
2.97


 


1020




460.19
Yes
3.3


 


1021




456.18
Yes
3.1


 


1022




428.18
Tentative
2.88


 


1023




428.18
Yes
3.01


 


1024




412.19
Tentative
2.86


 


1025




409.15
Yes
2.74


 


1026




520.13
No


 


1027




398.17
Tentative
2.93


 


1028




452.08
Yes
3.36


 


1029




462.07
Yes
3.2


 


1030




462.07
Yes
3.02


 


1031




462.07
Tentative
3.21


 


1032




418.12
Yes


 


1033




432.1
Yes
3.36, 6.43


 


1034




418.12
No


 


1035




452.08
Yes
3.53


 


1036




440.22
Yes
3.66


 


1037




452.08
Yes


 


1038




486.11
No


 


1039




476.2
Yes
3.21, 3.49,6.51


 


1040




412.19
Tentative
2.95


 


1041




444.18
Yes
2.84


 


1042




428.18
Tentative
2.74


 


1043




412.19
No


 


1044




409.2
Yes
2.75, 3.22


 


1045




418.1
Tentative
3.1, 3.5


 


1046




508.2
Yes
3.01, 3.39


 


1047




502.2
Yes
4.18, 4.67


 


1048




436.1
Yes
3.18, 3.61


 


1049




490.1
Yes
3.48, 3.99


 


1050




486.11
Yes
3.79


 


1051




486.1
Yes
2.79, 3.73,4.25


 


1052




486.11
Yes
3.33


 


1053




432.1
Tentative
3.16, 3.62


 


1054




478.14
Yes
3.09


 


1055




468.14
Yes
3.42


 


1056




436.1
Yes
3.27, 3.73


 


1057




446.2
Yes
3.36, 3.87


 


1058




494.15
Yes
3.73


 


1059




490.14
Yes
3.52


 


1060




462.2
Yes
3.24, 3.75


 


1061




462.2
Yes
3.43, 4.17


 


1062




446.2
Yes
3.21, 3.7


 


1063




443.11
Yes
3.08


 


1064




554.09
Yes
4.59


 


1065




432.1
Tentative
3.32, 3.82


 


1066




486.04
Yes
3.85


 


1067




496.03
Yes
3.65


 


1068




496.03
Yes
3.39


 


1069




496  
Yes
3.65, 4.15


 


1070




452.08
Yes
3.55


 


1071




466.1
Yes
3.83, 6.53


 


1072




452.08
Yes
3.37


 


1073




486.04
No


 


1074




474.18
Yes
4.14


 


1075




486.04
Yes
4.01


 


1076




520.07
Yes
4.15


 


1077




510.15
Yes
3.96


 


1078




446.2
Yes
3.31, 3.8


 


1079




478.1
Yes
3.2, 3.79


 


1080




462.2
Yes
3.04, 3.49


 


1081




446.15
No


 


1082




443.1
Yes
3.07, 3.47


 


1083




390.21
Tentative
3.1


 


1084




480.24
Yes
2.71, 3.05


 


1085




474.3
Yes
4.39


 


1086




408.2
Tentative
3.19


 


1087




462.23
Yes
3.5


 


1088




458.19
Yes
3.77


 


1089




458.19
Yes
2.7, 3.72


 


1090




458.19
Yes
3.38


 


1091




404.22
Tentative
3.21


 


1092




450.23
Yes
3.12


 


1093




440.22
Yes
3.48


 


1094




408.2
Tentative
3.27


 


1095




418.24
Yes
3.44


 


1096




466.24
Yes
3.81


 


1097




462.23
Yes
3.53


 


1098




434.23
Yes
3.28, 6.53


 


1099




434.23
Yes
3.45, 6.47


 


1100




418.24
Tentative
3.33


 


1101




415.2
Tentative
3.09


 


1102




526.18
Tentative
2.96


 


1103




404.22
Tentative
3.35


 


1104




458.13
Yes
2.7, 3.86


 


1105




468.12
No


 


1106




468.12
Yes
3.42


 


1107




468.12
Yes
3.64


 


1108




424.17
Yes
3.55


 


1109




438.18
Yes
3.83


 


1110




424.17
Yes
3.4


 


1111




458.13
Yes
4.01


 


1112




446.27
Yes
4.17


 


1113




458.13
Yes
2.7, 4.01


 


1114




492.15
No


 


1115




482.23
Yes
3.98


 


1116




418.24
No


 


1117




450.23
Yes
3.23, 6.51


 


1118




434.23
Tentative
3.15


 


1119




418.24
Tentative
3.48


 


1120




415.2
Yes
3.09


 







The additional examples below were prepared by individual syntheses.


 











1121




604.56
yes


 


1122




570.11
yes


 


1123




602-59
yes


 


1124




645.66
yes


 


1125




590.52
yes


 


1126




612.52
yes


 


1127




678.07
yes


 


1128




587.51
yes


 


1129




568.53
yes


 


1130




526.49
yes


 


1131




526.49
yes


 


1132




546.91
yes


 


1133




580.46
yes


 


1134




546.91
yes


 


1135




583.59
yes


 


1136




612.518
yes


 


1137




630.533
yes


 


1138




576.485
yes


 


1139




582.553
yes


 


1140




587.508
yes


 


1141




551.475
yes


 


1142




582.126
yes


 


1143




632.142
yes


 


1144




596.109
yes


 


1145




576.142
yes


 


1146




582.082
yes


 


1147




616.527
yes


 


1148




571.102
yes


 


1149




521.086
yes


 


1150




614.555
yes


 


1151




580.11
yes


 


1152




565.095
yes


 


1153




534.081
yes


 


1154




513.663
yes


 


1155




517.626
yes


 


1156




535.069
yes









The compounds listed in Table 1, or their pharmaceutically acceptable salts, may be used in the methods described herein to treat or prevent pain.
It will be understood that when compounds of the present invention contain one or more chiral centers, the compounds of the invention may exist in, and be isolated as, enantiomeric or diastereomeric forms, or as a racemic mixture. The present invention includes any possible enantiomers, diastereomers, racemates or mixtures thereof, of a compound of Formula I. The optically active forms of the compound of the invention may be prepared, for example, by chiral chromatographic separation of a racemate, by synthesis from optically active starting materials or by asymmetric synthesis based on the procedures described thereafter.
It will also be appreciated that certain compounds of the present invention may exist as geometrical isomers, for example E and Z isomers of alkenes. The present invention includes any geometrical isomer of a compound of Formula I. It will further be understood that the present invention encompasses tautomers of the compounds of the formula I.
It will also be understood that certain compounds of the present invention may exist in solvated, for example hydrated, as well as unsolvated forms. It will further be understood that the present invention encompasses all such solvated forms of the compounds of the formula I.
Within the scope of the invention are also salts of the compounds of the formula I. Generally, pharmaceutically acceptable salts of compounds of the present invention may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound, for example an alkyl amine with a suitable acid, for example, HCl or acetic acid, to afford a physiologically acceptable anion. It may also be possible to make a corresponding alkali metal (such as sodium, potassium, or lithium) or an alkaline earth metal (such as a calcium) salt by treating a compound of the present invention having a suitably acidic proton, such as a carboxylic acid or a phenol with one equivalent of an alkali metal or alkaline earth metal hydroxide or alkoxide (such as the ethoxide or methoxide), or a suitably basic organic amine (such as choline or meglumine) in an aqueous medium, followed by conventional purification techniques.
In one embodiment, the compound of formula I above may be converted to a pharmaceutically acceptable salt or solvate thereof, particularly, an acid addition salt such as a hydrochloride, hydrobromide, phosphate, acetate, fumarate, maleate, tartrate, citrate, methanesulphonate or p-toluenesulphonate.
The novel compounds of the present invention are useful in therapy, especially for the treatment of various pain conditions such as chronic pain, neuropathic pain, acute pain, cancer pain, pain caused by rheumatoid arthritis, migraine, visceral pain etc. This list should however not be interpreted as exhaustive.
Compounds of the invention are useful in disease states where degeneration or dysfunction of Bradykinin receptors is present or implicated in that paradigm. This may involve the use of isotopically labeled versions of the compounds of the invention in diagnostic techniques and imaging applications such as positron emission tomography (PET).
Compounds of the invention are useful for the treatment of septic shock, pancreatitis, edema, rhinitis, asthma, colitis, arthritis, hepatorenal syndrome, cancer, (including but not restricted to SCLC, prostrate cancer), bacterial and viral infections, ulcerative colitis, and Alzheimer's Disease.
Compounds of the invention are useful as an analgesic agent for use during general anesthesia and monitored anesthesia care. Combinations of agents with different properties are often used to achieve a balance of effects needed to maintain the anesthetic state (e.g. amnesia, analgesia, muscle relaxation and sedation). Included in this combination are inhaled anesthetics, hypnotics, anxiolytics, neuromuscular blockers and opioids.
Also within the scope of the invention is the use of any of the compounds according to the formula I above, for the manufacture of a medicament for the treatment of any of the conditions discussed above.
A further aspect of the invention is a method for the treatment of a subject suffering from any of the conditions discussed above, whereby an effective amount of a compound according to the formula I above, is administered to a patient in need of such treatment.
Thus, the invention provides a compound of formula I, or pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined for use in therapy.
In a further aspect, the present invention provides the use of a compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined in the manufacture of a medicament for use in therapy.
In the context of the present specification, the term “therapy” also includes “prophylaxis” unless there are specific indications to the contrary. The term “therapeutic” and “therapeutically” should be construed accordingly. The term “therapy” within the context of the present invention further encompasses to administer an effective amount of a compound of the present invention, to mitigate either a pre-existing disease state, acute or chronic, or a recurring condition. This definition also encompasses prophylactic therapies for prevention of recurring conditions and continued therapy for chronic disorders.
The compounds of the present invention are useful in therapy, especially for the therapy of various pain conditions including, but not limited to: acute pain, chronic pain, neuropathic pain, acute pain, back pain, cancer pain, and visceral pain.
In use for therapy in a warm-blooded animal such as a human, the compound of the invention may be administered in the form of a conventional pharmaceutical composition by any route including orally, intramuscularly, subcutaneously, topically, intranasally, intraperitoneally, intrathoracially, intravenously, epidurally, intrathecally, intracerebroventricularly and by injection into the joints.
In one embodiment of the invention, the route of administration may be orally, intravenously or intramuscularly.
The dosage will depend on the route of administration, the severity of the disease, age and weight of the patient and other factors normally considered by the attending physician, when determining the individual regimen and dosage level at the most appropriate for a particular patient.
For preparing pharmaceutical compositions from the compounds of this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories.
A solid carrier can be one or more substances, which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or table disintegrating agents; it can also be an encapsulating material.
In powders, the carrier is a finely divided solid, which is in a mixture with the finely divided compound of the invention, or the active component. In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
For preparing suppository compositions, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture in then poured into convenient sized moulds and allowed to cool and solidify.
Suitable carriers are magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like.
The term composition is also intended to include the formulation of the active component with encapsulating material as a carrier providing a capsule in which the active component (with or without other carriers) is surrounded by a carrier which is thus in association with it. Similarly, cachets are included.
Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral administration.
Liquid form compositions include solutions, suspensions, and emulsions. For example, sterile water or water propylene glycol solutions of the active compounds may be liquid preparations suitable for parenteral administration. Liquid compositions can also be formulated in solution in aqueous polyethylene glycol solution.
Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavoring agents, stabilizers, and thickening agents as desired. Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the pharmaceutical formulation art.
Depending on the mode of administration, the pharmaceutical composition will preferably include from 0.05% to 99% w (percent by weight), more preferably from 0.10 to 50% w, of the compound of the invention, all percentages by weight being based on total composition.
A therapeutically effective amount for the practice of the present invention may be determined, by the use of known criteria including the age, weight and response of the individual patient, and interpreted within the context of the disease which is being treated or which is being prevented, by one of ordinary skills in the art.
Within the scope of the invention is the use of any compound of formula I as defined above for the manufacture of a medicament.
Also within the scope of the invention is the use of any compound of formula I for the manufacture of a medicament for the therapy of pain.
Additionally provided is the use of any compound according to Formula I for the manufacture of a medicament for the therapy of various pain conditions including, but not limited to: acute pain, chronic pain, neuropathic pain, acute pain, back pain, cancer pain, and visceral pain.
A further aspect of the invention is a method for therapy of a subject suffering from any of the conditions discussed above, whereby an effective amount of a compound according to the formula I above, is administered to a patient in need of such therapy. Additionally, there is provided a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier.
Particularly, there is provided a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier for therapy, more particularly for therapy of pain. Further, there is provided a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier use in any of the conditions discussed above.
In a further aspect, the present invention provides a method of preparing a compound of formula I.
Methods of Preparation
The compounds listed in Table 1 were prepared as single compounds in a combinatorial array. The Table 1, column 4 designation of “yes” indicates that the target mass of the designated compound was found in >50% abundance in the MS spectrum. Similarly, the designation of “tentative” indicates that the target mass of the designated compound was found in 15-50% abundance in the MS spectrum. Likewise the designation “no” indicates that the target mass of the designated compound was found in <15% abundance in MS 'spectrum. It will be understood by those of ordinary skill in the art that a chemical reaction which fails to efficiently yield the desired product within the context of a combinatorial protocol may nonetheless efficiently yield the desired product when the reaction is performed in a single reaction or parallel reaction format, without undue experimentation on the part of the chemist. In this regard, several of the compounds which were not prepared efficiently in the combinatorial array, were subsequently prepared in separate syntheses as shown in the Examples.
The reaction sequence depicted in Scheme 1, infra, describes a process for preparing compounds of formula (I) wherein X is represented by formula (i) or (ii), comprising reacting a compound of general formula II




wherein Y is a protecting group such as CBZ or FMOC, with an alkyl or alkenyl halide, such as allyl bromide in the presence of a base such as cesium carbonate to give compounds of general formula III;





deprotecting the compounds of formula III under standard conditions and then acylated the deprotected products of formula IV with thiophosgene or phosgene to yield isothiocyanates or isocyanates of formula V, respectively:





and subsequently aminating the compounds of formula V combinatorially in a multiwell plate with a selection of different amines to yield compounds of formula I wherein X is represented by formula (i) or (ii) shown above.

The general protocol for the preparation of the combinatorial library is depicted in Scheme 1 and specific experimental details are provided in the Examples below.




    
    
        b is 0, 1, or 2; R1, R2, R3, R4, and R5 are as defined above.
    
    


In addition, certain compounds of the present invention and certain intermediates used in the preparation of the compounds of the present invention may be prepared according to one or more of the following general procedures, wherein, unless specified otherwise, b, R1, R2, R3, R4 and R5 are defined as above.




As illustrated in the scheme above, to a stirred solution of (7-chloro-2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-, phenylmethyl ester-carbamic acid (10 mmol) in DMF (140 ml) was added cesium carbonate (3.72 g, 11.4 mmol) followed by methyl iodide (2.0 g, 14 mmol). The reaction mixture was stirred at room temperature for 5 hours then concentrated in vacuo. The residue was taken in EtOAc (200 ml) and washed with brine (2×30 ml). The organic phase was then dried over MgSO4, filtered and concentrated in vacuo. The products were purified by flash chromatography using dichloromethane as the eluent. The products (8 mmol) were added with conc HBr (33% in acetic acid) (50 ml) and were stirred at room temperature for 3 hours. The reaction mixture was poured into ether (300 ml), the precipitate was collected, and then taken in dichloromethane (250 ml) and washed with 2N NaOH (2×50 ml). The organic phase was dried over MgSO4, filtered and concentrated in vacuo to provide the desired compound.




As illustrated in the scheme above, to a solution of 3-aminobenzodiazepines (3 mmol) in dichloroethane (20 ml) and saturated aqueous Na2CO3 (20 ml) was added dropwise thiophosgene (0.7 g, 6 mmol). The reaction mixture was stirred at room temperature for 3 hours. The organic phase was separated and the aqueous phase was extracted with dichloromethane (50 ml). The combined organic phases were dried over MgSO4, filtered and concentrated in vacuo. The product was purified by flash chromatography (100% CH2Cl2) to yield a compound of formula V.







As illustrated in the scheme above, to a solution of 4-fluoro-2-methyl-1-nitro-benzene (0.155 g, 1 mmol) in a 1:1 mixture of ethanol and water (20 ml) was added morpholine (0.435 g, 5 mmol). The reaction mixture was heated at 90° C. for 16 h. The solvent was evaporated in vacuo, the residue was taken in dichloromethane (50 ml) and washed with brine (3×10 ml). The organic phase was dried over MgSO4, filtered, and concentrated in vacuo. The product was taken up in ethanol (20 ml) and heated at reflux. A solution of tin chloride (2M, 2.5 ml) in conc. HCl was added dropwise and heated at reflux for another 30 minutes. The solvent was then evaporated in vacuo, and the residue was treated with a 2M solution of sodium hydroxide until the pH of the solution was >10. The mixture was extracted with dichloromethane (50 ml) and the organic phase was dried over MgSO4, filtered, and concentrated in vacuo. The product (0.032 g, 0.2 mmol) ((ESI) (M+H)+=192) was taken up in dichloroethane (5 ml), and 7-chloro-5-(2-chlorophenyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one(0.075 g, 0.2 mmol) was added. The reaction mixture was heated at 70 C for 16 h. The solvent was evaporated in vacuo, and the residue was washed with ether (2×10 ml) to yield the desired compound.




As illustrated in the scheme above, N2 gas was bubbled through both DME and water for at least 3 hours. A solution of the imidoyl chloride (1 equiv.) in DME (1.5 mL/mmol imidoyl chloride) was placed in a N2purged flask. Na2CO3 (1 equiv.), PdCl2(dppf) (0.05 equiv.), boronic acid (1 equiv.) and water (0.5 mL/mmol imidoyl chloride) were added sequentially, and the resulting mixture was heated at 100° C. until the imidoyl chloride was consumed (typically 16 h). The reaction was then cooled, diluted with CH2Cl2 and water, and the layers were separated. The aqueous phase was extracted with CH2Cl2 (3×), and the combined organic phases were dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography to provide the desired compound.
General Procedure 5:




As illustrated in the scheme above, boronic acid (1.15 equiv.), Pd2(dba)3 (0.015 equiv.), and dry KF (3.3 equiv.) were placed in an oven-dried, N2 purged flask. A solution of the imidoyl chloride (1 equiv.) in dry THE (2 mL/mmol imidoyl chloride) was added followed by a solution of P(t-Bu)3 (0.045 equiv., 10% solution in hexanes) in dry THF (1.6 mL/mmol imidoyl chloride). The resulting mixture was heated at reflux until the imidoyl chloride was consumed (typically 16 h). The reaction was then cooled, diluted with EtOAc, and filtered through a small pad of silica gel. The silica was washed well with EtOAc, and the combined organic phases were concentrated in vacuo. The product was purified by silica gel column chromatography to provide the desired compound.




As illustrated in the scheme above, a solution of the benzodiazepine compound (1 equiv.) in dry THF (4 mL/mmol benzodiazepine, or slightly more if solubility was low) was added to a mixture of KHMDS (1.05 equiv., 0.5 M in toluene) and dry THF (2 mL/mmol benzodiazepine) immersed in a −78° C. cooling bath. After stirring for 5 min., a solution of trisyl azide (2.5 equiv) in dry THF (4 mL/mmol benzodiazepine) was added to the reaction, and stirring was continued until all the starting benzodiazepine had been consumed (typically 10 min.). Glacial acetic acid (4.4 equiv) was then added, and the mixture was warmed to 30° C. for 2 h. Saturated NaHCO3 was added, the layers were separated, and the aqueous phase was extracted with CH2Cl2 (4×). The combined organic phases were dried over Na2SO4, filtered, and concentrated in vacuo. The product was purified by silica gel column chromatography to provide the corresponding azide compound.




As illustrated in the scheme above, polymer supported triphenylphosphine (Argonaut Technologies, 5-10 equiv) was added to a solution of the azide (1 equiv.) in THF (10 mL/g polymer) and water (0.8 mL/g polymer). The resulting mixture was stirred at room temperature until all of the azide had been consumed (typically overnight). The polymer resin was then removed by filtration, and was washed well with CH2Cl2 and MeOH (3× each). The filtrate was concentrated in vacuo, and the residue was redissolved in CH2Cl2. Any remaining water was removed with the aid of a separatory funnel, and the organic phase was dried over Na2SO4, filtered, and concentrated in vacuo. The amine was purified using a “catch and release” strategy with MP-TsOH resin (Argonaut Technologies): The product was dissolved in CH2Cl2 (10 mL/mmol product), and MP-TsOH resin (2.3 equiv) was added. The mixture was stirred for 1 h, and the solvent was removed by filtration and discarded. The resin was rinsed with CH2Cl2 and MeOH (3× each), and the washings were discarded as well. The product was then released from the resin by washing with 2M NH3 in MeOH and CH2Cl2 (3× each). Concentration of the filtrate in vacuo provided the corresponding amine compound.
Additional compounds of the present invention may also be prepared according to the methods represented in Schemes 2-4 below, wherein, unless specified otherwise, b, R1, R2, R3, R4 and R5 are defined as above.











Biological Evaluation
I. B2 Bradykinin Binding Assay
A. Human Bradykinin B2 (hB2) Receptor Expression and Membrane Preparation
The cloned human Bradykinin B2 (hB2) receptor in the pCIN vector was purchased from Receptor Biology. The hB2 receptor was stably transfected into HEK 293 S cells and a clonal cell line was generated. Cells were grown in T-flasks with DMEM culture media containing 10% FBS, 2 mM glutamine, 600 μg/ml neomycin and an antibiotic cocktail (100 IU penicillin, 100 μg/ml streptomycin, 0.25 μg/ml amphotericin B). Membranes, expressing the hB2 receptor, were prepared from this cell line according to this protocol: Cells are harvested at 1 to 1.2 million cells/ml, pelleted, and resuspended in ice-cold lysis buffer (50 mM Tris, pH 7.0, 2.5 mM EDTA, with PMSF added just prior to use to 0.5 mM from a 0.5 M stock in DMSO. After lysis on ice for 15 min, the cells are homogenized with a polytron for 10 sec. The suspension is spun at 1000 g for 10 min at 4° C. The supernatant is saved on ice and the pellets resuspended and spun as before. The supernatants from both spins are combined and spun at 46,000 g for 10-30 min. The pellets are resuspended in cold Tris buffer (50 mM Tris/Cl, pH 7.0) at a dilution of 0.2-1 ml per 40 million cells and spun again. The final pellets are resuspended in membrane buffer (50 mM Tris, 0.32 M sucrose, pH 7.0). Aliquots are frozen in dry ice/ethanol and stored at −70° C. until use. The protein concentrations are determined by a modified Lowry with SDS.
B. hB2 Receptor Binding
Membranes expressing the hB2 receptor are thawed at 37° C., passed 3 times through a 25-gauge blunt-end needle, diluted in the bradykinin binding buffer (50 mM Tris, 3 mM MgCl2, and 1 mg/ml BSA, pH 7.4, 0.02 mg/ml Phenanthroline, 0.25 mg/ml Pefabloc) and 80 μL aliquots containing the appropriate amount of protein (final concentration of 0.25 μg/ml) are distributed in 96-well polystyrene plates (Treff Lab). The IC50 of compounds are evaluated from 10-point dose-response curves, where the serial dilutions are done on a final volume of 150 μL, with 70 μL of 125I-Desamino-TyrHOE140 (Kd=0.05) at 50,000 to 60,000 dpm per well (0.03-0.04 nM) in a final volume of 300 μl. The total and non-specific binding are determined in the absence and presence of 0.1 μM (150 μL) of Bradykinin respectively. The plates are vortexed and incubated for 60 minutes at room temperature, filtered through Unifilters-96 GF/B (Canberra Packard), which were presoaked in 0.1% polyethyleneimine, with a harvester using 3 ml of wash buffer (50 mM Tris, pH 7.0, 3 mM MgCl2). The filters are dried for 1 hour at 55° C. The radioactivity (cpm) is counted in a TopCount (Canberra Packard) after adding 65 μl/well of MS-20 scintillation liquid (Canberra Packard). Compounds of the present invention have demonstrated hB2 receptor binding at concentrations less than 10 μM.
Based on the above assays, the dissociation constant (Ki) for a particular compound of the invention towards a particular receptor is determined using the following equation:

Ki=IC50/(1+[rad]/Kd),

Wherein IC50 is the concentration of the compound of the invention at which 50% displacement has been observed;
[rad] is a standard or reference radioactive ligand concentration at that moment; and
Kd is the dissociation constant of the radioactive ligand towards the particular receptor.
II. GTP[γ]35S Binding Experiments on Bradykinin (B2) Receptors
A. General Information
The procedures below describe how to perform and interpret GTP[γ]35S binding experiments designed to determine the activity of new compounds on the human B2 receptor.
B. General Procedure of the Assay
Human Bradykinin-2 GTP[γ]35S Binding
Human Bradykinin-2 membranes (hB2 293s) are thawed at 37° C., passed 3 times through a 25-gauge blunt-end needle and diluted in the GTPγS binding buffer for the assay (50 mM Hepes, pH 7.4; 200 mM NaCl; 1 mM EDTA; 5 mM MgCl2. To this added freshly prepared 1 mM DTT, 0.5% BSA, 1 μM GDP. The EC50 and Emax of compounds are evaluated from 10-point dose-response curves done in 300 μl with the appropriate amount of membrane protein and 100,000-120,000 dpm of GTPγ35S per well (0.11-0.14 nM). Bradykinin (1-9) is used as the standard agonist at hB2. The ranges of concentrations tested should include a maximal concentration of 0.1 μM bradykinin in order to establish the Emax.
The plates are vortexed and incubated for 60 minutes at room temperature, filtered on GF/B Unifilters (presoaked in water) with the Packard harvester using 4 ml/well of wash buffer (50 mM Tris, 5 mM MgCl2, 50 mM NaCl, pH 7.0), minimum. The filters are dried for 1 hour at 55° C. The radioactivity (cpm) is counted in a TopCount (Packard) after adding 65 μl/well of MS-20 scintillation liquid.
Antagonist reversal studies are done in a similar manner except that the compound dose-response curve's are performed in the presence of a constant concentration of agonist (approx. 80% bradykinin Emax;  ̃5 nM). A standard B2 Antagonist is used as the reference antagonist at hB2. The ranges of antagonist concentrations tested should include a maximal concentration of 3 μM of the standard B2 Antagonist in order to establish the maximal displacement (Dmax).
C. Radioligand: Preparation of GTP[γ]35S
GTP[γ]35S is acquired from Perkin-Elmer (250 μCi/20 μl). It is diluted from with 10 mM DTT, 50 mM Tris, pH 7 (dilute in 2 ml, 1.0 mCI/20 μ). Sonicate the solution, filter through a 0.45 μm filter, and freeze aliquots at −70° C. For the experiment, use  ̃0.3 nM dilution of this tracer in the GTP binding buffer.
D. Data Analysis
The EC50 and Emax of compounds are evaluated from 10-point dose-response curves done in 300 μl with the appropriate amount of membrane protein and GTPγ35S per well and are calculated in Activity base with ExcelFit. The basal and maximal stimulated binding are determined in the absence and presence of standard reference compounds, respectively.
The stimulation (Stim) in the presence of compounds is expressed as the percentage of Dmax of the reference antagonist. Values of IC50, Ki′ and Dmax for ligands capable of competing for agonist stimulated binding are calculated in Activity Base. Mean±S.E.M. values of IC50, Ki′ and % Dmax are reported for ligands tested in at least three dose-response curves.
Biological data for particular testing samples (as listed in Table 2) of the compounds of the invention are listed in Table 3 below.







TABLE 2







List of the test samples used in the Biological Evaluation








Test



Sample Nos:
Structure of the Test Sample











1






 


2






 


3






 


4






 


5






 


6






 


7






 


8






 


9






 


10






 


11






 


12






 


13






 


14






 


15






 


16






 


17






 


18






 


19






 


20






 


21






 


22






 


23






 


24






 


25






 


26






 


27






 


28






 


29






 


30






 


31






 


32






 


33




















TABLE 3







Biological Data for the testing samples as listed in Table 2










Test Sample Nos.
Ki (hB2) (nM)







1-33
43-3110










EXAMPLES
The invention will further be described in more detail by the following Examples which describe methods whereby compounds of the present invention may be prepared, purified, analyzed and biologically tested, and which are not to be construed as limiting the invention.
Intermediate 1
3-amino-7-chloro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one




Following General Procedure 1, INTERMEDIATE 1 was obtained as pale brown solid (2.5 g, 77%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)+=300.
Intermediate 2
3-amino-5-(2-bromophenyl)-7-fluoro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one




Following General procedure 1, INTERMEDIATE 2 was obtained as a thick pale brown oil (0.5 g, 17%) and used for the subsequent reaction without further purification.
Intermediate 3
3-amino-5-cyclohexyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one




Following General Procedure 1, after flash chromatography (100% EtOAc), INTERMEDIATE 3 was obtained as a pale brown solid (1.25 g, 45%). MS (ESI) (M+H)+=272
Intermediate 4
3-amino-7-chloro-5-(2-chlorophenyl)1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one




Following General Procedure 1, INTERMEDIATE 4 was obtained as a thick pale brown oil (1.8 g, 65%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)+=334
Intermediate 5
3-amino-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one




To a stirred solution of (2,3-dihydro-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-, phenylmethyl ester-carbamic acid, (4.0 g, 10.3 mmol) in toluene (100 ml) was added Aliquat336 (1.0 g) and 50% aqueous sodium hydroxide (20 ml) followed by methyl iodide (5.0 g, 35 mmol). The reaction mixture was stirred at room temperature for 17 hours. The solvent was evaporated in vacuo and the residue was taken in dichloromethane (150 ml). The organic phase was washed with 2N sodium hydroxide (50 ml) and brine (50 ml); the organic phase was dried over MgSO4, filtered and concentrated in vacuo. The residue was triturated with hexane and the precipitate was treated with conc. HBr as in General Procedure 1. INTERMEDIATE 5 was obtained as a pale brown solid (1.63 g, 59%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)+=266
Intermediate 6
3-amino-7-chloro-1,3-dihydro-5-phenyl-1-(2-propenyl)-2H-1,4-benzodiazepin-2-one




To a stirred solution of (7-chloro-2,3-dihydro-2-oxo-5-phenyl-1-H-1,4-benzodiazepin-3-yl)-, phenylmethyl ester-carbamic acid (1 mmol) in DMF (10 ml) was added cesium carbonate (0.370 g, 1.14 mmol) followed by allyl bromide (0.134 g, 1.1 mmol). The reaction mixture was stirred at room temperature for 5 hours. The solvent was evaporated in vacuo and the residue was taken in EtOAc (50 ml). The organic phase was washed with brine (2×10 ml); the organic phase was dried over MgSO4, filtered and concentrated in vacuo. The crude products were purified by flash chromatography using dichloromethane as the eluent. The crude product was added to conc. HBr (33% in acetic acid) (10 ml) and stirred at room temperature for 3 hours. The reaction mixture was poured into ether (100 ml), the precipitate was collected and then taken in dichloromethane (50 ml) and washed with 2N NaOH (2×10 ml). The organic phase dried over MgSO4, filtered and concentrated in vacuo. The title compound was obtained as a pale yellow solid (0.19 g, 60%) and used for the subsequent reaction without further purification. MS (ESI) (M+H)+=326
Intermediate 7
7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one




Following General Procedure 2, INTERMEDIATE 7 was obtained as a pale yellow solid (0.7 g, 69%). 1H-NMR (CDCl3): δ 7.66-7.64 (m, 2H), 7.60-7.57 (dd, J=2.4 Hz and 8.8 Hz, 1H), 7.55-7.51 (m, 1H), 7.47-7.43 (m, 2H), 7.36-7.34 (m, 2H) and 3.48 (s, 4H) MS (ESI) (M+H)+=342
Intermediate 8
7-chloro-5-(2-chlorophenyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one




Following General Procedure 2, after flash chromatography (dichloromethane), the title compound was obtained as pate yellow solid (2.2 g, 65%). 1H-NMR (CDCl3): δ 7.66-7.64 (m, 1H), 7.56-7.53 (dd, J=2.4 Hz and 8.8 Hz, 1H), 7.46-7.43 (m, 2H), 7.39-7.37 (m, 1H), 7.05 (d, J=2.0 Hz, 1H), 3.73 (s, 1H) and 3.51 (s, 3H). MS (ESI) (M+H)+=376
Example 1
N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-(5isoquinolinyl)-thiourea








As illustrated in the scheme above, to a solution of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 7) (0.035 g, 0.1 mmol) in dichloromethane (5 ml) was added 5-isoquinolinamine (0.015 g, 0.1 mmol). The reaction mixture was heated at reflux for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (20 mg, 40%). 1H-NMR (CDCl3): δ 9.33 and 9.19 (2×s, 1H), 8.63 and 8.49 (2×d, J=5.6 Hz and J=6.0 Hz, 1H), 8.34 (br s, 1H), 8.02 (d, J=8.4 Hz, 1H), 7.92 (t, J=11.6 Hz, 2H), 7.58-7.32 (m, 10H), 6.03 (d, J=7.6 Hz, 1H) and 3.3 (s, 3H). MS (ESI) (M+H)+=486
Example 2
N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4(dimethylamino)phenyl]-thiourea




To a solution of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 7) (0.342 g, 1 mmol) in dichloromethane (20 ml) was added N,N-dimethyl-1,4-benzenediamine (0.136 g, 1 mmol). The reaction mixture was heated at reflux for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (0.167 g, 35%). 1H-NMR (CDCl3): δ 7.70-7.22 (m, 10H), 6.75 (d, J=8.8 Hz, 2H), 6.05 (d, J=7.6 Hz, 1H), 3.42 (s, 3H) and 2.98 (s, 6H). MS (ESI) (M+H)+=478.
Example 3
N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4(dimethylamino)-1-naphthalenyl]-N-methyl-thiourea




To a stirred solution of carboxybenzyl-3-amino-7-chloro-5-phenyl-2-oxo-1,4-benzodiazepine (EXAMPLE 6) (1 mmol) in DMF(10 ml) was added cesium carbonate (5 mmol) followed by methyl iodide (2.2 mmol). The reaction mixture was stirred at room temperature for 5 hours. The solvent was evaporated in vacuo and the residue was taken in EtOAc (50 ml). The organic phase was washed with brine (2×10 ml); the organic phase was dried over MgSO4, filtered and concentrated in vacuo. The product (0.8 mmol) was added to conc. HBr (35% in acetic acid) (10 ml) and was stirred at room temperature for 3 hours. The reaction mixture was poured into ether (30 ml), the precipitate was collected, and taken in dichloromethane (25 ml) and washed with 2N NaOH (2×10 ml). The organic phase was dried over MgSO4, filtered and concentrated in vacuo. The product was used for the subsequent steps without further purification.
To a solution of the product above (0.03 g, 1 mmol) in dichloroethane (5 ml) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.025 g, 1 mmol). The reaction mixture was stirred at 70 C for 4 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (28 mg, 52%). 1H-NMR (CDCl3): δ 8.24 (m, 1H), 7.92 (m, 1H), 7.67 (d, J=7.2 Hz, 2H), 7.52-7.40 (m, 8H), 7.31 (d, J=1.6 Hz, 1H), 7.05 (d, J=8.0 Hz, 2H), 3.78 (s, 3H), 3.41 (s, 3H) and 2.91 (s, 6H). MS (ESI) (M+H)+=542.
Example 4
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea




To a solution of 3-amino-7-chloro-5-(2-chlorophenyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 8) (0.03 g, 1 mmol) in dichloroethane (5 ml) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.025 g, 1 mmol). The reaction mixture was stirred at 70° C. for 4 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (35 mg, 60%). 1H-NMR (CDCl3): δ 8.26 (d, J=8.4 Hz, 1H), 8.03 (d, J=7.6 Hz, 1H), 7.93 (br s, 1H), 7.73-7.06 (m, 11H) 6.10 (d, J=7.6 Hz, 1H), 3.40 (s, 3H) and 2.92 (s, 6H). MS (ESI) (M+H)+=562.
Example 5
N-[7-chloro-2,3-dihydro-2-oxo-5-phenyl-1-(2-propenyl)-1H-1,4-benzodiazepin-3-yl]-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea




A solution of 3-amino-7-chloro-1,3-dihydro-5-phenyl-1-(2-propenyl)-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 6) (0.162 g, 0.5 mmol) in dichloroethane (5 ml) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.115 g, 0.5 mmol). The reaction mixture was stirred at 70° C. for 4 hours; The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless, solid (0.152 g, 55%). 1H-NMR (CDCl3): δ 8.29-8.27 (m, 1H), 8.08-8.06 (m, 1H), 7.90 (s, 1H), 7.60-7.36 (m, 10H), 7.32 (d, J=2.4 Hz, 1H), 7.09 (d, J=8 Hz, 1H), 6.14 (d, J=7.6 Hz, 1H), 5.76-5.69 (m, 1H), 5.15 (s, 1H), 5.12 (dd, J=1.2 and 7.0 Hz, 1H), 4.57-4.39 (m, 2H) and 2.93 (s, 6H). MS (ESI) (M+H)+=554.
Example 6
N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea




To a solution of 3-amino-7-chloro-5-phenyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 1) (0.299 g, 1 mmol) in dichloroethane (15 mi) was added 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.230 g, 1 mmol). The reaction mixture was stirred at room temperature for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (0.345 g, 65%). 1H-NMR (CD3OD): δ 8.25 (d, 8.4 Hz, 2H), 8.00 (d, J=8.4 Hz, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.85-7.81 (m, 2H), 7.77-7.72 (m, 2H), 7.65 (d, J=8.8 Hz, 1H), 7.61-7.58 (m, 4H), 7.51-7.47 (m, 2H), 7.29 (d, J=2.4 Hz, 1H), 6.01 (s, 1H) and 3.49 (s, 9H). MS (ESI) (M+H)+=528.
Example 7
N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-(4-methoxy-2-methylphenyl)-urea




To a solution of 3-amino-7-chloro-5-phenyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (INTERMEDIATE 1) (0.299 g, 1 mmol) in dichloromethane (15 ml) was added 1-isocyanato-4-methoxy-2-methyl-benzene (0.163 g, 1 mmol). The reaction mixture was stirred at room temperature for 17 hours. The solvent was evaporated in vacuo, the residue was triturated with ether and the title compound was obtained as a colorless solid (0.197 g, 42%). 1H-NMR (CDCl3): δ 7.57-7.48 (m, 3H), 7.47-7.44 (m, 1H), 7.39-7.38 (m, 3H), 7.36-7.29 (m, 2H), 6.77-6.73 (m, 2H), 6.71 (d, J=2.8 Hz, 1H), 6.61 (s, 1H), 5.51 (d, J=8.4 Hz, 1H), 3.78 (s, 3H), 3.39 (s, 3H) and 2.29 (s, 3H). MS (ESI) (M+H)+=463.
Example 8
N-(7-chloro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[4-(dimethylamino)-1-naphthalenyl]-urea








As illustrated in the scheme above, a mixture of 4-(dimethylamino)-1-naphthalenecarboxylic acid (0.1 mmol), diphenylphosphoryl azide (0.15 mmol) and triethylamine (0.3 mmol) in toluene (10 ml) was heated at reflux overnight. The solvent was evaporated in vacuo, and then the residue was redissolved in dichloroethane, added to INTERMEDIATE 1 (0.08 mmol) and heated at 70° C. for 4 hours. The solvent was evaporated in vacuo and the residue was taken in dichloromethane (50 ml). The organic phase was washed with brine (2×10 ml); the organic phase was dried over MgSO4, filtered and concentrated in vacuo. The residue was triturated with ether and the title compound was obtained as a colorless solid (9 mg, 22%); 1H-NMR (CDCl3): δ 8.27 (m, 1H), 8.11 (m, 1H), 7.60 (d, J=8 Hz, 1H), 7.77-7.29 (m, 10H), 7.08 (d, J=8.4 Hz, 1H), 6.75 (br s, 2H), 5.56 (d, J=8.4 Hz, 1H), 3.39 (s, 3H) and 2.90 (s, 6H); MS (ESI) (M+H)+=512.
Example 9
N-[5-(2-bromophenyl)-7-fluoro-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-1-naphthalenyl]-thiourea




As illustrated in the scheme above, INTERMEDIATE 2 (0.018 g, 0.05 mmol) and 4-isothiocyanato-N,N-dimethyl-1-naphthalenamine (0.011 g, 0.05 mmol) was heated in dichloroethane (4 mL) at 70° C. overnight. The solvent was evaporated in vacuo and the residue was trituated with ether (2×10 ml). The title compound (0.021 g, 72%) was obtained as a colorless solid. 1H-NMR (CDCl3): δ 8.26 (m, 1H), 8.06 (m, 1H), 7.85 (s, 1H), 7.79 (m, 1H), 7.69-7.24 (m, 8H), 7.08 (d, J=8.0 Hz, 1H), 7.02 (t, J=8.4 Hz, 1H), 6.06 (d, J=8.0 Hz, 1H), 3.39 (s, 3H), 2.93 (s, 6H). (ESI) (M+H)+=591.
Example 10
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′′-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]-guanidine (E and Z isomers Separated but not Identified)








As illustrated in the scheme above, a solution of N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]-thiourea (EXAMPLE 11, made according to General Procedure 3) (0.058 g, 0.1 mmol) and silver triflate (0.077 g, 0.3 mmol) in dichloromethane (2 ml) was combined with cyanamide disodium salt (1 mmol) and stirred at room temperature for 3 h. The reaction mixture was then diluted with dichloromethane (10 ml) and washed with brine (2×5 ml). The organic phase was dried over MgSO4, filtered and concentrated in vacuo. The residue was washed with ether (2×10 ml) and the polar isomer of the title compound (0.023 g, 380%) was obtained as pale yellow solid. The ether layer was concentrated in vacuo and purified by column chromatography (1:1 EtOAc:CH2Cl2) to give the non-polar isomer of the title compound (0.009 g, 15%) as a colorless solid.
Example 10A
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′′-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]-guanidine (polar isomer)
1H-NMR (CDCl3): δ 8.26 (m, 1H), 7.9 (m, 2H), 7.69 (d, J=12.0 Hz, 2H), 7.53-7.46 (m, 4H), 7.39 (m, 1H), 7.30-7.23 (m, 4H), 7.11 (m, 2H), 3.98 (t, J=9.2 Hz, 2H), 3.48 (s, 3H) and 3.12 (br.s, 6H). (ESI) (M+H)+=612
Example 10B
N-[7-chloro-5(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′′-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]-guanidine (nonpolar isomer)
1H-NMR (CDCl3): δ 8.25 (m, 1H), 8.03 (m, 1H), 7.66-7.53 (m, 5H), 7.44-7.40 (m, 3H), 7.39-7.30 (m, 2H), 7.09 (m, 2H), 6.54 (d, J=7.6 Hz, 1H), 5.48 (d, J=7.6 Hz, 1H), 3.99 (t, J=9.2 Hz, 4H), 3.39 (s, 3H) and 3.15 (br.s, 4H). (ESI) (M+H)+=612.
Example 11
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]-thiourea




Following General Procedure 3, the title compound (0.078 g, 65%) was obtained as a white solid. 1H-NMR (CDCl3): δ 8.4 (m, 1H), 8.08 (m, 2H), 7.72 (m, 1H), 7.64 (d, J=8 Hz, 1H), 7.6-7.5 (m, 4H), 7.39 (t, J=4.4 Hz, 2H), 7.30 (d, J=8.8 Hz, 2H), 7.10 (m, 2H), 6.09 (d, J=7.6 Hz, 1H), 3.98 (t, J=4.4 Hz, 4H) 3.40 (s, 3H), and 3.13 (m, 4H). (ESI) (M+H)+=604.
Example 12
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4(4-morpholinyl)-1-naphthalenyl]-guanidine




As illustrated in the scheme above, to a solution of N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]-thiourea (EXAMPLE 11) (0.029 g, 0.05 mmol) and silver triflate (0.038 g, 0.15 mmol) in dichloromethane (2 ml) was added a solution of ammonia (0.25 ml, 2M in methanol). The reaction mixture was stirred at room temperature for 3 h, diluted with dichloromethane (10 ml) and washed with brine (2×5 ml). The organic layer was dried with MgSO4 filtered, concentrated in vacuo. The residue was triturated with ether to give the title compound (0.008 g, 28%) as a pale yellow solid. 1H-NMR (CDCl3): δ 8.22 (d, J=9.2 Hz, 1H), 8.11 (d, J=8.4 Hz, 1H), 7.75 (br.s, 1H), 7.50-7.41 (m, 7H), 7.31 (d, J=8.8 Hz, 1H), 7.04 (m, 2H), 5.8 (br.s, 1H), 3.96 (t, J=9.2 Hz, 4H) 3.51 (s, 3H) and 3.05 (br.s, 4H).). (ESI) (M+H)+=587.
Example 13
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[2-methyl-4-(4-morpholinyl)phenyl]-thiourea




Following General Procedure 3, the title compound (0.096 g, 85%) was obtained as a colorless solid. 1H-NMR (CDCl3): δ 7.74 (m, 1H), 7.54 (s, 1H), 7.51 (m, 1H), 7.42-7.39 (m, 3H), 7.34-7.31 (m, 2H), 7.24 (d, J=8.8 Hz, 1H), 7.09 (d, J=2.4 Hz, 1H), 6.80-6.76 (m, 2H), 6.09 (d, J=7.6 Hz, 1H), 3.85 (t, J=9.6 Hz, 4H), 3.44 (s, 3H), 3.17 (t, J=10.0 Hz, 4H) and 2.32 (s, 3H). (ESI) (M+H)+=568.
Example 14
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-2-methylphenyl]-thiourea




Following the General Procedure 3, the title compound (0.052 g, 51%) was obtained as a pale brown solid. 1H-NMR (CDCl3): δ 7.75 (m, 1H), 7.52 (dd, J=8.8 Hz, 1H), 7.47 (s, 1H), 7.42-7.40 (m, 3H), 7.34-7.31 (m, 2H), 7.19 (d, J=8.0 Hz, 1H), 7.09 (d, J=2.0 Hz, 1H), 6.60 (br.s, 1H), 6.58 (br.s, 1H), 6.09 (d, J=8.0 Hz, 1H), 3.44 (s, 3H), 2.97 (s, 6H) and 2.30 (s, 3H). (ESI) (M+H)+=526.
Example 15
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-3-methylphenyl]-thiourea




Following General Procedure 3, the title compound (0.034 g, 35%) was obtained as a pale brown solid. 1H-NMR (CDCl3): δ 7.86 (d, J=7.6 Hz, 1H), 7.76 (m, 1H), 7.53 (dd, J=8.8 Hz, 1H), 7.42-7.39 (m, 2H), 7.36-7.32 (m, 2H), 7.17 (dd, J=8.8 Hz, 1H), 7.12 (d, J=2.0 Hz, 1H), 7.10 (d, J=2.4 Hz, 1H), 7.06 (d, J=8.4 Hz, 1H), 6.09 (d, J=7.6 Hz, 1H), 3.46 (s, 3H), 2.71 (s, 6H) and 2.33 (s, 3H). (ESI) (M+H)+=526.
Example 16
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[3-chloro-4-dimethylamino)phenyl]-thiourea




Following the General Procedure 3, the title compound (0.025 g, 23%) was obtained as a pale brown solid. 1H-NMR (CDCl3): δ 7.86 (s, 1H), 7.83 (d, J=7.2 Hz, 1H), 7.5 (m, 1H), 7.53 (dd, J=8.8 Hz, 1H), 7.43-7.40 (m, 2H), 7.35-7.33 (m, 2H), 7.28 (m, 1H), 7.10 (s, J=2.0 Hz, 1H), 7.05 (d, J=7.2 Hz, 1H), 6.06 (d, J=7.2 Hz, 1H), 3.47 (s, 3H) and 2.82 (s, 6H). (ESI) (M+H)+=546.
Example 17
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(dimethylamino)-3-(trifluoromethyl)phenyl]-thiourea




Following General Procedure 3, the title compound (0.017 g, 18%) was obtained as a pale brown solid. 1H-NMR (CDCl3): δ 7.93 (s, 1H), 7.85 (d, J=7.2 Hz, 1H), 7.38 (m, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.55-7.53 (m, 2H), 7.42-7.40 (m, 2H), 7.34 (m, 3H), 7.10 (s, J=2.0 Hz, 1H), 6.06 (d, J=7.6 Hz, 1H), 3.47 (s, 3H) and 2.76 (s, 6H). (ESI) (M+H)+=580.
Example 18
N-[7-chloro-5(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-chloro-2-(dimethylamino)phenyl]-thiourea




Following the General Procedure 3, the title compound (0.024 g, 22%) was obtained as a pale brown solid. 1H-NMR (CDCl3): δ 8.55 (d, J=7.2 Hz, 1H), 7.84 (s, 1H), 7.77 (t, J=4.8 Hz, 1H), 7.53 (dd, J=8.8 Hz, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.42-7.41 (m, 2H), 7.36-7.34 (m, 3H), 7.11-7.02 (m, 2H), 60.6 (d, J=7.2 Hz, 1H), 3.48 (s, 3H) and 2.75 (s, 6H). (ESI) (M+H)+=546.
Example 19
N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzo-diazepin-3-yl]-N′-[4-(diethylamino)-2-(dimethylamino)phenyl]-thiourea




Following the General Procedure 3, the title compound (0.040 g, 35%) was obtained as a pale gray solid. 1H-NMR (CDCl3): δ 9.4 (s, 1H), 8.22 (s, 1H), 7.76 (m, 1H), 7.52 (dd, J=8.8 Hz, 1H), 7.40 (m, 2H), 7.35 (m, 2H), 7.10 (d, J=2.4 Hz, 1H), 7.03 (d, J=8.8 Hz, 1H), 6.84 (s, 1H), 6.49 (dd, J=8.8 Hz, 1H), 6.11 (d, J=7.2 Hz, 1H), 3.47 (s, 3H) 3.33 (m, 4H), 2.65 (s, 6H) and 1.38 (t, J=14.0 Hz, 6H). ESI) (M+H)+=583.
Example 20
N-[(1E)-[[7-Chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]amino][[4-(4-morpholinyl)-1-naphthalenyl]amino]methylene]urea




A mixture of N-[7-chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′′-cyano-N′-[4-(4-morpholinyl)-1-naphthalenyl]guanidine (EXAMPLE 10) (21.9 mg, 35.8 μmol), water (4.6 μL, 260 μmol) and trifluoroacetic acid (19.8 μL, 257 μmol) in THF (3 mL) was heated to reflux for 43 h. The reaction was concentrated in vacuo, and the residue was purified by reverse phase HPLC (gradient 20-70% CH3CN in H2O) to provide the title compound (0.0092 g, 35%) as its TFA salt. 1H-NMR (CD3OD): δ 8.37 (br s, 1H), 8.06 (br s, 1H), 7.75-7.41 (br m, 9H), 7.26 (br s, 1H), 7.04 (br s, 1H), 5.66 (br s, 1H), 4.00 (br s, 4H), 3.58 (br s, 3H), 3.17 (br s, 4H). HRMS calculated for (C32H29Cl2N7O3+H) (M+H)+: 630.1787. Found (ESI): 630.1800.
Example 21
N′-[7-Chloro-5-(2-chlorophenyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N-methyl-N-[2-methyl-4-(4-morpholinyl)phenyl]thiourea




A solution of 2-methyl4-(4-morpholinyl)benzenamine (53.3 mg, 0.277 mmol) and diisopropylethylamine (0.063 mL, 0.36 mmol) in CH2Cl2 (1 mL) was cooled to 0° C. Methyl chloroformate (0.024 mL, 0.31 mmol) was added dropwise, and then the reaction was allowed to warm to room temperature and stir overnight. The reaction was diluted with CH2Cl2 (20 mL) and washed with brine (10 mL). The organic phase was dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was then suspended in a 1:2 mixture of Et2O:THF (6 mL). A solution of LiAlH4 in Et2O (0.34 mL of a 1 M solution, 0.34 mmol) was added dropwise, and then the reaction mixture was heated to reflux for 1.5 h. The reaction was cooled, diluted with additional Et2O (8 mL), and quenched with Na2SO45H2O (0.98 g, 4.2 mmol). After stirring for 15 minutes, the mixture was filtered and the reaction was concentrated in vacuo. A portion of this crude aniline (0.0580 g, 0.281 mmol) was dissolved in (CH2Cl)2 (8 mL), and 7-chloro-5-(2-chlorophenyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one (0.106 g, 0.281 mmol) was added. The resulting mixture was heated at 70° C. for 14 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (7:1 CH2Cl2:EtOAc) to provide the title compound (0.1211 g, 74%). Due to hindered rotation about one of the bonds, rotamers were observed in the 1H-NMR spectrum. 1H-NMR (CDCl3): δ 7.77-7.68 (m, 1H), 7.51 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.43-7.36 (m, 2H), 7.34-7.28 (m, 2H), 7.16 (t, J=8.4 Hz, 1H), 7.10-7.01 (m, 2H), 6.86-6.76 (m, 2H), 6.14 and 6.10 (2×d, J=8.0 Hz, J=7.6 Hz, 1H), 3.88-3.82 (br m, 4H), 3.60 and 3.59 (2×s, 3H), 3.41 (s, 3H), 3.19 (br s, 4H), 2.24 and 2.23 (2×s, 3H). MS (ESI) (M+H)+=582. HRMS calculated for (C29H29Cl2N5O2S+H) (M+H)+: 582.1497. Found (ESI): 582.1448.
Intermediate 9
6-Chloro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione




As illustrated in the scheme above, NaH (2.43 g of a 60% dispersion, 60.8 mmol) was added to a solution of 6-chloro-2H-3,1-benzoxazine-2,4(1H)-dione (10.0 g, 50.6 mmol) dissolved in DMF (200 mL). The resulting mixture was stirred at room temperature for 30 min., and then methyl iodide (6.3 mL, 101 mmol) was added dropwise. After the reaction had stirred at room temperature overnight, it was concentrated in vacuo. Water and brine were added to the residue, and the aqueous layer was extracted with CH2Cl2 (2×). The combined organic phases were dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was triturated with 3:1 hexanes:EtOAc. The solvent was removed by filtration, and the resulting solid was washed with additional 3:1 hexanes:EtOAc, followed by 100% hexanes. The product was dried briefly under vacuum to produce the title compound as a pale yellow solid. (8.59 g, 80%). 1H-NMR (DMSO-d6): δ 7.96 (d, J=2.6 Hz, 1H), 7.89 (dd, J=2.6 Hz, J=9.0 Hz, 1H), 7.48 (d, J=9.0 Hz, 1H), 3.45 (s, 3H). MS (ESI) (M+H)+=212.
Intermediate 10
7-Chloro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione




As illustrated in the scheme above, a mixture of 6-chloro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione (6.00 g, 28.4 mmol) and glycine (2.14 g, 28.4 mmol) in glacial acetic acid (72 mL) was heated at reflux for 4 h. The reaction was cooled and concentrated in vacuo. Water was added to the residue, and the mixture was cooled to 0° C. NaHCO3 was added to adjust the pH of the aqueous layer to approximately 8, and then the aqueous layer was extracted with CH2Cl2 (3×). The combined organic phases were dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was triturated with Et2O, the solvent was removed by filtration, and the resulting solid was washed with additional Et2O to provide the title compound as a slightly yellow solid (5.657 g, 89%). 1H-NMR (CDCl3): δ 7.88 (d, J=2.5 Hz, 1H), 7.57 (br s, 1H), 7.53 (dd, J=2.6 Hz, J=8.7 Hz, 1H), 7.19 (d, J=8.6 Hz, 1H), 3.84 (d, J=6.1 Hz, 2H), 3.39 (s, 3H). MS (ESI) (M+H)+=225.
Intermediate 11
5,7-Dichloro-1,3-dihydro-1-methyl-2H-1,4benzodiazepin-2-one




As illustrated in the scheme above, 7-chloro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione (5.00 g, 22.3 mmol) was suspended in POCl3 (100 mL) and heated at 100° C. for 30 min. The reaction was cooled and concentrated in vacuo. Traces of POCl3 were removed by adding toluene and concentrating the mixture in vacuo (2×). The residue was dissolved in CH2Cl2, the solution was cooled to 0° C., and Et3N (6.8 mL, 48.8 mmol) was added dropwise. The mixture was stirred for 1 h and allowed to slowly warm to room temperature, and was then concentrated in vacuo once again. The residue was purified by silica gel column chromatography (5:1 CH2Cl2:EtOAc+0.5% Et3N) to provide the title compound as an orange solid (4.68 g, 86%). 1H-NMR (CDCl3): δ 7.79 (d, J=2.5 Hz, 1H), 7.55 (dd, J=2.4 Hz, J=8.9 Hz, 1H), 7.23 (d, J=8.8 Hz, 1H), 4.67 (br s, 1H), 3.72 (br s, 1H), 3.39 (s, 3H). MS (ESI) (M+H)+=243.
Intermediate 12
7-Chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 5, (2,4-dimethoxy-5-pyrimidinyl)boronic acid (0.939 g, 5.10 mmol), Pd2(dba)3 (0.064 g, 0.07 mmol), dry KF (0.890 g, 15.3 mmol), 5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.12 g, 4.61 mmol) and P(t-Bu)3 (0.42 mL of a 10% solution in hexanes, 0.21 mmol) were combined and heated for 20 h. After workup, purification of the crude product by silica gel column chromatography (1:3 hexanes:EtOAc) provided the title compound as a pale orange solid (1.20 g, 75%). 1H-NMR (CDCl3): δ 8.50 (s, 1H), 7.49 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.29 (d, J=8.8 Hz, 1H), 7.13 (d, J=2.4 Hz, 1H), 4.84 (d, J=10.8 Hz, 1H), 4.06 (s, 3H), 3.79 (s, 3H), 3.76 (d, J=10.8 Hz, 1H), 3.42 (s, 3H). MS (ESI) (M+H)+=347.
Intermediate 13
3-Azido-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 6, KHMDS (7.0 mL of 0.5 M in toluene, 3.5 mmol), 7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.16 g, 3.35 mmol), trisyl azide (2.60 g, 8.40 mmol) and acetic acid (0.85 mL, 14.8 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (2:3 hexanes:EtOAc) provided the title compound as a pale yellow solid (1.23 g, 95%). 1H-NMR (CDCl3): δ 8.64 (s, 1H), 7.54 (dd, J=2.3 Hz, J=8.8 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 7.19 (d, J=2.3 Hz, 1H), 4.51 (s, 1H), 4.08 (s, 3H), 3.78 (s, 3H), 3.47 (s, 3H). MS (ESI) (M+H)+388.
Intermediate 14
3-Amino-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 7, 3-azido-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.22 g, 3.15 mmol) and PS—PPh3 (23.0 g of 1.37 mmol/g, 31.5 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a brown solid (1.15 g, quantitative). 1H-NM (CDCl3): δ 8.56 (s, 1H), 7.50 (dd, J=2.4 Hz, J=8.9 Hz, 1H), 7.30 (d, J=8.8 Hz, 1H), 7.15 (d, J=2.5 Hz, 1H), 4.46 (s, 1H), 4.06 (s, 3H), 3.77 (s, 3H), 3.46 (s, 3H), 2.85-2.12 (br s, 2H). MS (ESI) (M+H)+=362.
Intermediate 15
7-Chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 2, 3-amino-7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.15 g, 3.18 mmol) and thiophosgene (0.49 mL, 6.4 mmol) were combined. Purification of the crude product by silica gel column chromatography (9:1 CH2Cl2:EtOAc) provided the title compound as a viscous dark yellow oil (0.560 g, 44%). 1H-NMR (CDCl3): δ 8.60 (s, 1H), 7.55 (dd, J=2.5 Hz, J=8.8 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 7.17 (d, J=2.5 Hz, 1H), 5.16 (s, 1H), 4.07 (s, 3H), 3.78 (s, 3H), 3.49 (s, 3H). MS (ESI) (M+H)+=404.
Example 22
N-[7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[4-(diethylamino)-2-methylphenyl]-thiourea




As illustrated in the scheme above, a solution of 7-chloro-5-(2,4-dimethoxy-5-pyrimidinyl)-1,3-dihydro-3-isothiocyanato-1-methyl-2H-1,4-benzodiazepin-2-one (0.069 g, 0.146 mmol) and N4,N4-diethyl-2-methyl-1,4benzenediamine (0.029 g, 0.161 mmol) in (CH2Cl)2 (5.0 mL) was heated at 70° C. for 16 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (7:3 CH2Cl2:EtOAc) to provide the title compound as a light orange solid (0.074 g, 87%). 1H-NMR (CDCl3): δ 8.64 (s, 1H), 7.52 (dd, J=2.5 Hz, J=8.8 Hz, 1H), 7.40 (s, 1H), 7.35 (d, J=7.8 Hz, 1H), 7.32 (d, J=9.0 Hz, 1H), 7.20 (d, J=2.2 Hz, 1H), 7.17-7.13 (m, 1H), 6.54-6.51 (m, 2H), 6.06 (d, J=8.0 Hz, 1H), 4.05 (s, 3H), 3.74 (s, 3H), 3.42 (s, 3H), 3.40-3.27 (m, 4H), 2.30 (s, 3H), 1.17 (t, J=7.1 Hz, 6H). HRMS calculated for (C28H32ClN7O3S+H) (M+H)+: 582.2054. Found (ESI): 582.2076.
Intermediate 16
7-Chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 5,3-thienylboronic acid (1.15 g, 9.01 mmol), Pd2(dba)3 (0.113 g, 0.123 mmol), dry KF (1.57 g, 27.0 mmol), 5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.98 g, 8.15 mmol) and P(t-Bu)3 (0.75 mL of a 10% solution in hexanes, 0.37 mmol) were combined and heated for 16 h. After workup, purification of the crude product by silica gel column chromatography (9:1 CH2Cl2:EtOAc) provided the title compound as a yellow solid (1.44 g, 61%). 1H-NMR (CDCl3): δ 7.54-7.49 (m, 4H), 7.37 (dd, J=3.0 Hz, J=5.0 Hz, 1H), 7.29 (d, J=9.4 Hz, 1H), 4.76 (d, J=10.9 Hz, 1H), 3.77 (d, J=10.9 Hz, 1H), 3.38 (s, 3H). MS (ESI) (M+H)+=291.
Intermediate 17
3-Azido-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 6, KHMDS (10.4 mL of 0.5 M in toluene, 5.20 mmol), 7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (1.44 g, 4.95 mmol), trisyl azide (3.83 g, 12.4 mmol) and acetic acid (1.25 mL, 21.8 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (7:3 hexanes:EtOAc) provided the title compound as a pale yellow solid (1.60 g, 98%). 1H-NMR (CDCl3): δ 7.61-7.55 (m, 4H), 7.40 (dd, J=2.9 Hz, J=5.1 Hz, 1H), 7.33 (dd, J=1.0 Hz, J=8.2 Hz, 1H), 4.54 (s, 1H), 3.44 (s, 3H). MS (ESI) (M+H)+=332.
Intermediate 18
3-Amino-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 7, 3-azido-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (1.60 g, 4.82 mmol) and PS—PPh3 (30.0 g of 1.37 mmol/g, 41.1 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a brown solid (1.35 g, 92%). 1H-NMR (CDCl3): δ 7.56-7.50 (m, 4H), 7.36 (dd, J=3.1 Hz, J=4.9 Hz, 1H), 7.29 (d, J=8.8 Hz, 1H), 4.46 (s, 1H), 3.43 (s, 3H), 2.35-2.15 (br s, 2H). MS (ESI) (M+H)+=306.
Intermediate 19
7-Chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 2,3-amino-7-chloro-1,3-dihydro-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (1.35 g, 4.41 mmol) and thiophosgene (0.67 mL, 8.8 mmol) were combined. Purification of the crude product by silica gel column chromatography (100% CH2Cl2) provided the title compound as a yellow solid (1.04 g, 68%). 1H-NMR (CDCl3): δ 7.61-7.54 (m, 4H), 7.39 (dd, J=2.9 Hz, J=5.3 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 5.20 (s, 1H), 3.46 (s, 3H). MS (ESI) (M+H)+=348.
Example 23
N-[7-Chloro-2,3-dihydro-1-methyl-2-oxo-5-(3-thienyl)-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]thiourea




This reaction was carried out in a multiwell plate. As illustrated in the scheme above, aA mixture of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-thienyl)-2H-1,4-benzodiazepin-2-one (156 μL of a 0.128 M solution in (CH2Cl)2, 0.020 mmol), 4-(4-morpholinyl)-1-naphthalenamine (44 μL of a 0.5 M solution in DMA, 0.022 mmol), and (CH2Cl)2 (300 μL) was agitated and heated at 70° C. for 22 h. The reaction was cooled and concentrated in vacuo, and the residue was redissolved in DMA (25 μL) and (CH2Cl)2 (275 μL). Polyamine resin HL (NovaBiochem) was added (20 mg of 4.53 mmol/g, 0.091 mmol), and the mixture was agitated at room temperature overnight. The resin was removed by filtration and washed with additional (CH2Cl)2 and MeOH. The filtrate was concentrated in vacuo to provide the title compound. MS (ESI) (M+H)+=576.
Intermediate 20
7-Chloro-1,3dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 4,5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.99 g, 8.19 mmol), Na2CO3 (0.868 g, 8.19 mmol), PdCl2(dppf) (0.335 g, 0.410 mmol) and (3-pyridinyl)boronic acid (1.01 g, 8.19 mmol) were combined and heated for 14 h. After workup, purification of the crude product by silica gel column chromatography (100% EtOAc) provided the title compound (1.51 g, 64%). 1H-NMR (CDCl3): δ 8.78 (d, J=1.6 Hz, 1H), 8.72 (dd, J=1.6 Hz, J=4.8 Hz, 1H), 8.02 (dt, J=1.6 Hz J=8.0 Hz, 1H),7.56 (dd, J=2.4 Hz, J=8.8 Hz,1H), 7.39 (dd, J=4.8 Hz, J=8.0 Hz, 1H), 7.34 (d, J=8.8 Hz, 1H), 7.28 (d, J=2.4 Hz, 1H), 4.89 (d, J=10.4 Hz, 1H), 3.80 (d, J=10.8 Hz, 1H), 3.41 (s, 3H); MS (ESI) (M+H)+=286.
Intermediate 21
3-Azido-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 6, KHMDS (10.5 mL of 0.5 M in toluene, 5.25 mmol), 7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one (1.43 g, 4.99 mmol), trisyl azide (3.86 g, 12.5 mmol) and acetic acid (1.26 mL, 22.0 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (1:1 CH2Cl2:EtOAc) provided the title compound as a yellow foam (1.47 g, 90%). 1H-NMR (CDCl3): δ 8.80 (s, 1H), 8.76 (d, J=3.6 Hz, 1H), 8.14 (dt, J=1.6 Hz, J=8.0 Hz, 1H), 7.61 (dd, J=2.0 Hz, J=8.4 Hz, 1H), 7.44 (ddd, J=0.8 Hz, J=4.8 Hz, J=8.0 Hz, 1H), 7.38 (d, J=8.8 Hz, 1H), 7.35 (d, J=2.4 Hz, 1H), 4.56 (s, 1H), 3.48 (s, 3H). MS (ESI) (M+H)+=327.
Intermediate 22
3-Amino-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 7, 3-azido-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one (1.47 g, 4.49 mmol) and PS—PPh3 (16.4 g of 1.37 mmol/g, 22.4 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a slightly brown solid (1.46 g, quantitative). 1H-NMR (CDCl3): δ 8.76 (d, J=2.4 Hz, 1H), 8.72 (dd, J=1.6 Hz, J=5.2 Hz, 1H), 8.06 (dt, J=2.4 Hz, J=7.6 Hz, 1H), 7.58 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.40 (dd, J=4.8 Hz, J=8.0 Hz, 1H), 7.35 (d, J=8.8 Hz, 1H), 7.29 (d, J=2.4 Hz, 1H), 4.51 (s, 1H), 3.46 (s, 3H), 2.41 (br s, 2H). MS (ESI) (M+H)+=301.
Intermediate 23
7-Chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above, a solution of 3-amino-7-chloro-1,3-dihydro-1-methyl-5-(3-pyridinyl)-2H-1,4benzodiazepin-2-one (1.36 g, 4.52 mmol) in dry THF (55 mL) was cooled to −15° C. Carbon disulfide (2.7 mL, 45 mmol) was added, followed by EDCI (1.73 g, 9.03 mmol). The mixture was stirred for 10 min., and then Et3N (1.26 mL, 9.04 mmol) was added. The reaction was stirred for 16 h while it was allowed to slowly warm to room temperature. The precipitated solid was removed by filtration and was washed well with CH2Cl2 and then discarded. The filtrate was concentrated in vacuo, and the residue was dissolved in CH2Cl2. The organic phase was washed with water, saturated NaHCO3, and brine, and was then dried over Na2SO4, filtered, and concentrated in vacuo. Purification of the crude product by silica gel column chromatography (3:1 CH2Cl2:EtOAc) provided the title compound as a solid (0.652 g, 42%). 1H-NMR (CDCl3): δ 8.75 (s, 2H), 8.12 (dt, J=2.0 Hz, J=8.0 Hz, 1H), 7.63 (dd, J=2.0 Hz, J=8.8 Hz, 1H), 7.43 (dd, J=4.8 Hz, J=8.0 Hz, 1H), 7.39 (d, J=8.8 Hz, 1H), 7.33 (d, J=2.4 Hz, 1H), 5.22 (s, 1H), 3.50 (s, 3H). MS (ESI) (M+H)+=343.
Example 24
N-[7-Chloro-2,3-dihydro-1-methyl-2-oxo-5-(3-pyridinyl)-1H-1,4-benzodiazepin-3-yl]-N′-[4-(4-morpholinyl)-1-naphthalenyl]thiourea




A solution of 7-chloro-1,3-dihydro-3-isothiocyanato-1-methyl-5-(3-pyridinyl)-2H-1,4-benzodiazepin-2-one (0.0282 g, 0.0823 mmol) and 4-(4-morpholinyl)-1-naphthalenamine (0.0188 g, 0.0823 mmol) in (CH2Cl)2 (2.5 mL) was heated at 70° C. for 24 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (1:2 CH2Cl2:EtOAc) to provide the title compound (0.0263 g, 56%). 1H-NMR (CDCl3): δ 8.72-8.68 (m, 2H), 8.29-8.25 (m, 1H), 8.11-8.07 (m, 1H), 8.05-8.00 (m, 2H), 7.64-7.55 (m, 5H), 7.38-7.33 (m, 3H), 7.15 (d, J=8.0 Hz, 1H), 6.08 (d, J=7.6 Hz, 1H), 4.03-3.96 (br m, 4H), 3.39 (s, 3H), 3.15 (br s, 4H). HRMS calculated for (C30H27ClN6O2S+H) (M+H)+: 571.1683. Found (ESI): 571.1699. Anal. Calcd for C30H27ClN6O2S+0.7 H2O: C, 61.73; H, 4.90; N, 14.40. Found: C, 61.93;H, 4.79; N, 13.87.
Intermediate 24
6-Fluoro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione




As illustrated in the scheme above, NaH (0.312 g of a 60% dispersion, 7.80 mmol) was added to a solution of 6-fluoro-2H-3,1-benzoxazine-2,4(1H)-dione (1.176 g, 6.49 mmol) dissolved in DMF (60 mL). The resulting mixture was stirred at room temperature for 30 min., and then methyl iodide (0.81 mL, 13 mmol) was added dropwise. After the reaction had stirred at room temperature overnight, it was concentrated in vacuo. Water and brine were added to the residue, and the aqueous layer was extracted with CH2Cl2 (3×). The combined organic phases were dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was triturated with Et2O. The solvent was removed by filtration, and the resulting solid was washed with additional Et2O. The product was dried briefly under vacuum to produce the title compound as a white solid (1.00 g, 79%). 1H-NMR (CDCl3): δ 7.86-7.82 (m, 1H), 7.55-7.48 (m, 1H), 7.22-7.17 (m, 1H), 3.61 (s, 3H). MS (ESI) (M+H)+=196.
Intermediate 25
7-Fluoro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione




As illustrated in the scheme above, a mixture of 6-fluoro-1-methyl-2H-3,1-benzoxazine-2,4(1H)-dione (1.00 g, 5.12 mmol) and glycine (0.385 g, 5.13 mmol) in glacial acetic acid (13 mL) was heated at reflux for 4 h. The reaction was cooled and concentrated in vacuo. Water was added to the residue, and the mixture was cooled to 0° C. NaHCO3 was added to adjust the pH of the aqueous layer to approximately 8, and then the aqueous layer was extracted with CH2Cl2 (4×). The combined organic phases were dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was triturated with Et2O, the solvent was removed by filtration, and the resulting solid was washed with additional Et2O to provide the title compound as a slightly yellow solid (0.560 g, 52%). 1H-NMR (DMSO-d6): δ 8.80 (br s, 1H), 7.49-7.38 (m, 3H), 3.76 (br d, 1H), 3.47 (br d, 1H), 3.26 (s, 3H). MS (ESI) (M+H)+=209.
Intermediate 26
5-Chloro-7-fluoro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above, 7-fluoro-3,4-dihydro-1-methyl-1H-1,4-benzodiazepine-2,5-dione (0.495 g, 2.38 mmol) was suspended in POCl3 (10.6 mL) and heated at 100° C. for 30 min. The reaction was cooled and concentrated in vacuo. Traces of POCl3 were removed by adding toluene and concentrating the mixture in vacuo (2×). The residue was dissolved in CH2Cl2, the solution was cooled to 0° C., and Et3N (0.75 mL, 5.4 mmol) was added dropwise. The mixture was stirred for 0.5 h and allowed to slowly warm to room temperature, and was then concentrated in vacuo once again. The residue was purified by silica gel column chromatography (9:1 CH2Cl2:EtOAc+0.5% Et3N) to provide the title compound as a light tan solid (0.422 g, 78%). 1H-NMR (CDCl3): δ 7.53-7.48 (m, 1H), 7.34-7.24 (m, 2H), 4.66 (br s, 1H), 3.72 (br s, 1H), 3.39 (s, 3H). MS (ESI) (M+H)+=227.
Intermediate 27
7-Fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 4, 5-chloro-7-fluoro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (0.201 g, 0.887 mmol), Na2CO3 (0.0940 g, 0.887 mmol), PdCl2(dppf) (0.0362. g, 0.0443 mmol) and phenylboronic acid (0.108 g, 0.886 mmol) were combined and heated for 13 h. After workup, purification of the crude product by silica gel column chromatography (3:1 CH2Cl2:EtOAc) provided the title compound (0.180 g, 76%). 1H-NMR (CDCl3): δ 7.64-7.60 (m, 2H), 7.51-7.45 (m, 1H), 7.44-7.39 (m, 2H), 7.34 (dd, J=4.8 Hz, J=9.2 Hz, 1H), 7.31-7.25 (m, 1H), 7.02 (dd, J=2.8 Hz, J=8.8 Hz, 1H), 4.84 (d, J=10.8 Hz, 1H), 3.78 (d, J=10.8 Hz, 1H), 3.40 (s, 3H). MS (ESI) (M+H)+=269.
Intermediate 28
3-Azido-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 6, KHMDS (1.34 mL of 0.5 M in toluene, 0.670 mmol), 7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (0.171 g, 0.637 mmol), trisyl azide (0.493 g, 1.59 mmol) and acetic acid (0.16 mL, 2.8 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (100% CH2Cl2 to 9:1 CH2Cl2:EtOAc) provided the title compound as a pale yellow solid (0.163 g, 83%). 1H-NMR (CDCl3): δ 7.72-7.67 (m, 2H), 7.54-7.49 (m, 1H), 7.47-7.42 (m, 2H), 7.39 (dd, J=4.8 Hz, J=9.2 Hz, 1H), 7.36-7.30 (m, 1H), 7.08 (dd, J=2.8 Hz, J=8.4 Hz, 1H), 4.55 (s, 1H), 3.45 (s, 3H). MS (ESI) (M+H)+=310.
Intermediate 29
3-Amino-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 7, 3-azido-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (0.0826 g, 0.267 mmol) and PS—PPh3 (1.63 g of 1.64 mmol/g, 2.67 mmol) were combined. After workup and purification by “catch and release,” the title compound was obtained as a slightly yellow solid (0.0450 g, 59%). 1H-NMR (CDCl3): δ 7.65-7.60 (m, 2H), 7.51-7.46 (m, 1H), 7.44-7.39 (m, 2H), 7.36 (dd, J=4.8 Hz, J=9.2 Hz, 1H), 7.33-7.26 (m, 1H), 7.03 (dd, J=2.4 Hz, J=8.4 Hz, 1H), 4.48 (s, 1H), 3.45 (s, 3H), 2.30-1.60 (br s, 2H). MS (ESI) (M+H)+=284.
Example 25
N-(7-Fluoro-2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N′-[2-methyl-4-(4-morpholinyl)phenyl]thiourea




As illustrated in the scheme above, a solution of 3-amino-7-fluoro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (0.0056 g, 0.020 mmol) and 4-(4-isothiocyanato-3-methylphenyl)morpholine (0.0048 g, 0.020 mmol) in DMA (0.5 mL) was heated at 70° C. for 16 h. The reaction was cooled and concentrated in vacuo, and the residue was lyophilized to provide the title compound (0.0104 g, quantitative). 1H-NMR (CDCl3): δ 7.63-7.58 (m, 2H), 7.50-7.45 (m, 1H), 7.44-7.36 (m, 4H), 7.36-7.26 (m, 3H), 7.09 (dd, J=2.4 Hz, J=8.4 Hz, 1H), 6.85-6.78 (m, 2H), 6.09-6.05 (m, 1H), 3.90-3.83 (br m, 4H), 3.41 (s, 3H), 3.21-3.15 (br s, 4H), 2.36 (s, 3H). MS (ESI) (M+H)+=518.HRMS calculated for (C28H28FN5O2S+H) (M+H)+: 518.2026. Found (ESI): 518.2117.
Intermediate 30
7-Chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above, absolution of 2-bromo-6-methoxypyridine (0.022 mL, 0.18 mmol) in dry Et2O (0.3 mL) was added to a solution of n-BuLi (0.12 mL of 1.6 M in hexanes, 0.19 mmol) maintained at −40° C. The reaction was stirred at −40° C. for 20 min., and then B(OMe)3 (0.022 mL, 0.19 mmol) was added dropwise. The reaction was stirred at −40° C. for 30 min., and then at room temperature for 3.5 h. The reaction was concentrated in vacuo, anhydrous MeOH was added to the residue, and the reaction was concentrated in vacuo once again. Dry DME (0.8 mL), 5,7-dichloro-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (0.0392 g, 0.161 mmol), Pd(PPh3)4 (0.0093 g, 0.0080 mmol) and CsF (0.0612 g, 0.403 mmol) were added to the residue, and the mixture was heated to reflux for 15 h. Water (5 mL) and CH2Cl2 (5 mL) were added to the reaction mixture, and the layers were separated. The aqueous phase was extracted with additional CH2Cl2 (3×), and the combined organic phases were dried over Na2SO4, filtered, and concentrated in vacuo. Purification of the crude product by silica gel column chromatography (2:1 CH2Cl2:EtOAc) provided the title compound (0.029 g, 57%). 1H-NMR (CDCl3): δ 7.77 (d, J=7.6 Hz, 1H), 7.71-7.64 (m, 2H), 7.49 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.27 (d, J=8.8 Hz, 1H), 6.83 (d, J=8.0 Hz, 1H), 4.85 (d, J=10.8 Hz, 1H), 3.86 (d, J=10.4 Hz, 1H), 3.81 (s, 3H), 3.39 (s, 3H). MS (ESI) (M+H)+=316.
Intermediate 31
3-Azido-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above and following General Procedure 6, KHMDS (0.19 mL of 0.5 M in toluene, 0.095 mmol), 7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one (0.0290 g, 0.0918 mmol), trisyl azide (0.0710 g, 0.229 mmol) and acetic acid (0.023 mL, 0.40 mmol) were combined. After workup, purification of the crude product by silica gel column chromatography (49:1 CH2Cl2:EtOAc) provided the title compound (0.0204 g, 62%). 1H-NMR (CDCl3): δ 7.95 (d, J=7.2 Hz, 1H), 7.78-7.70 (m, 2H), 7.54 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.31 (d, J=8.8 Hz, 1H), 6.87 (d, J=8.4 Hz, 1H), 4.64 (s, 1H), 3.80 (s, 3H), 3.44 (s, 3H). MS (ESI) (M+H)+=357.
Intermediate 32
3-Amino-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one




As illustrated in the scheme above, a suspension of Pd/C (0.005 g of 10% on C) in MeOH (0.5 mL) under N2 was treated with ammonium formate (0.0252 g, 0.400 mmol). The mixture was stirred for 10 min. and then transferred via Pasteur pipette to a suspension of 3-azido-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one (0.0204 g, 0.0572 mmol) in MeOH (1.2 mL). The resulting mixture was stirred at room temperature for 3 h and was then filtered through a small pad of Celite. The filtrate was concentrated in vacuo, and the residue was purified by silica gel column chromatography (100% EtOAc, followed by 4:1 CH2Cl2:EtOAc) to provide the title compound (0.0148 g, 78%). 1H-NMR (CDCl3): δ 7.82 (dd, J=0.8 Hz, J=7.6 Hz, 1H), 7.71-7.66 (m, 2H), 7.51 (dd, 2.4 Hz, J=8.8 Hz, 1H), 7.28 (d, J=8.8 Hz, 1H), 6.83 (dd, J=0.8 Hz, J=8.4 Hz, 1H), 4.56 (s, 1H), 3.80 (s, 3H), 3.44 (s, 3H), 2.53 (br s, 2H). MS (ESI) (M+H)+=331.
Example 26
N-[7-Chloro-2,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl]-N′-[2-methyl-4-(4-morpholinyl)phenyl]thiourea




As illustrated in the scheme above, a solution of 3-amino-7-chloro-1,3-dihydro-5-(6-methoxy-2-pyridinyl)-1-methyl-2H-1,4-benzodiazepin-2-one (0.0113 g, 0.0342 mmol) and 4-(4-isothiocyanato-3-methylphenyl)morpholine (0.0080 g, 0.034 mmol) in (CH2Cl)2 (0.9 mL) was heated at 70° C. for 20 h. The reaction was cooled and concentrated in vacuo, and the residue was purified by silica gel column chromatography (2:1 CH2Cl2:EtOAc) to provide the title compound (0.0172 g, 89%) as a slightly yellow solid. 1H-NMR (CDCl3): δ 7.79 (dd, J=0.8 Hz, J=7.2 Hz, 1H), 7.70-7.65 (m, 2H), 7.54 (dd, J=2.4 Hz, J=8.8 Hz, 1H), 7.42 (br s, 1H), 7.30 (d, J=8.8 Hz, 1H), 7.28-7.25 (m partially hidden under CHCl3, 2H), 6.85-6.80 (m, 3H), 6.14 (m, 1H), 3.88-3.85 (m, 4H), 3.78 (s, 3H), 3.40 (s, 3H), 3.22-3.18 (m, 4H), 2.36 (s, 3H). MS (M+H+=565.

## Claims

1. A compound of Formula (I), or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof:




wherein
R1 is selected from optionally substituted acyl, optionally substituted alkyloxycarbonyl, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted cycloalkyl, optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, optionally substituted carbazolyl, optionally substituted heterocyclyl, optionally substituted aryl-C1-6alkyl, and optionally substituted heterocyclyl-C1-6alkyl;
X is —NHC(═S)N(R2)—;
R3 is substituted aryl, optionally substituted C1-12alkyl, optionally substituted C3-12cycloalkyl, or optionally substituted heterocyclyl;
R4 is, at each position, independently halogen, optionally substituted alkyl, optionally substituted heteroalkyl, nitro, cyano, hydroxy, —OR6, —SR6, —S(═O)R6, —S(═O)2R6, —C(═O)R6, —C(═S)R6, —NR7R6, —C(═O)NR7R6, —NR7C(═O)R6, —SO2NR7R6, —NR7SO2R6, or —C(═O)OR6;
R5, R6 and R7 are independently —H, or optionally substituted C1-6alkyl; and
R2 is selected from —H, optionally substituted C1-12alkyl, optionally substituted C1-12heteroalkyl, substituted aryl, and optionally substituted heterocyclyl.

2. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof wherein:
R1 is optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, or optionally substituted carbazolyl, wherein said naphthyl, isoquinolyl, acridinyl, coumarinyl, and carbazolyl are optionally substituted by C1-6alkyl , C1-6heterocyclyl or amino;
R2 is —H or C1-3alkyl;
R3 is substituted aryl, optionally substituted heteroaryl or optionally substituted cycloalkyl;
R4 is halogen, or C1-3alkyl; and
R5 is C1-3alkyl.

3. The compound of claim 2, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein:
R1 is optionally substituted naphthyl or optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino;
R2 is —H or C1-3alkyl;
R3 is optionally substituted cyclohexyl, optionally substituted pyridyl, optionally substituted thienyl, or optionally substituted pyrimidinyl, wherein said cyclohexyl, pyridyl, thienyl, and pyrimidinyl are optionally substituted by halogen, methoxy, or C1-3alkyl;
R4 is halogen; and
R5is methyl.

4. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein:
R1 is optionally substituted naphthyl or, optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino;
R2 is C1-3alkyl;
R3 is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C1-3alkyl;
R4 is halogen; and
R5is methyl.

5. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein R5 is —H or substituted C1-6alkyl.

6. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein:
R1 is optionally substituted naphthy or optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino;
R2 is —H or C1-3alkyl;
R3 is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C1-3alkyl;
R4 is halogen; and
R5 is substituted C1-3alkyl.

7. A compound of Formula I, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof:




wherein:
R1 is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, optionally substituted acridinyl, optionally substituted coumarinyl, optionally substituted carbazolyl, or a first divalent group selected from optionally substituted C1-12alkylene and optionally substituted C1-12heteroalkylene; wherein said naphthyl, isoquinolyl, acridinyl, coumarinyl, and carbazolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino, wherein said C1-12alkylene and C1-12heteroalkylene are optionally substituted by C1-6alkyl, aryl-C1-6alkyl, aryl or heterocyclyl;
X is —NHC(═S)N(R2)—;
R2 is —H, C1-3alkyl, or a second divalent group selected from a single bond, an optionally substituted alkylene and an optionally substituted heteroalkylene;
wherein said second divalent group together with said first divalent group forms a portion of a ring;
R3 is substituted aryl, optionally substituted heteroaryl or optionally substituted cycloalkyl;
R4 is halogen, or C1-3alkyl; and
R5 is C1-3alkyl.

8. The compound of claim 7, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein:
R1 is optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said naphthyl, and isoquinolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino;
R1 is —H or C1-3alkyl;
R3 is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C1-3alkyl;
R4 is halogen; and
R5 methyl.

9. The compound of claim 1, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof, wherein:
R1 is optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said naphthyl and isoquinolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino;
R2 is —H or C1-3alkyl;
R3 is optionally substituted cyclohexyl, substituted phenyl, substituted pyridyl, substituted thienyl, or substituted pyrimidinyl, wherein said cyclohexyl is optionally substituted by halogen, methoxy, or C1-3alkyl;
R4 is halogen; and
R5 methyl.

10. A mixture of two or more compounds selected from a compound of claim 1, a pharmaceutically acceptable salt thereof, a diasteromer thereof, and an enantiomer thereof.

11. A compound of Formula I, or pharmaceutically acceptable salt, diastereomer, or enantiomer thereof:




wherein;
—X—R1, in combination, is selected from —NHC(═S)N(R2)(R1) and




R1 is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolyl, wherein said phenyl, naphthyl and isoquinolyl are optionally substituted by C1-6alkyl, C1-6heterocyclyl or amino;
R2 is —H, or C1-3alkyl;





is a nitrogen containing heterocyclyl, which is optionally substituted by one or more —R8, and which includes a bond on the nitrogen that links to other group of formula (I);
R8 is —H, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted C1-6alkyl, —OH, or C1-6alkoxy, wherein R8 is optionally fused with the ring of




R3 is optionally substituted C3-12cyclohexyl, substituted phenyl, optionally substituted pyridyl, optionally substituted thienyl, or optionally substituted pyrimidinyl, wherein said C3-12cyclohexyl, phenyl, pyridyl, thienyl and pyrimidinyl are optionally substituted by halogen, methoxy, or C1-3alkyl;
R4 is halogen; and
R5 is methyl.

12. A compound as claimed in claim 11, wherein said nitrogen containing heterocyclyl is selected from piperazinyl, morpholinyl, piperidyl, and pyrrolidinyl.