CZ20003672A3 - Heterocyclic glycyl-beta-alanine derivatives as vitronectin agonists - Google Patents

Abstract

The present invention relates to a class of compounds represented by formula (I) or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing such compounds and methods of treating conditions mediated by the alpha v beta 3 integrin.

Description

Technical field

The invention relates to pharmaceutical agents (compounds) which are useful as αν β3 integrin antagonists and thus are useful in the pharmacological compositions and methods of treating conditions caused by _α ν β3 inhibit or antagonním effect of integrin _α ν β3.

BACKGROUND OF THE INVENTION

Integrins are a family of cell surface glycoproteins that allow cell adhesion and serve as cell mediators of cell adhesion interactions that occur during various cellular processes. Integrins are heterodimers consisting of non-covalently linked α and β polypeptide subunits. So far, eleven different α subunits and six different β subunits have been found. Different α subunits may be combined with different β subunits to form different integrins.

Integrins called α _ν β3 (also known as the vitronectin receptor) are considered to be integrins involved in various disease states or diseases including tumor metastasis, solid tumor growth (neoplasia), osteoporosis, Paget's disease, humoral hypercalcemia in malignancies, angiogenesis , including tumor angiogenesis, retinopathy, including macular degeneration, arthritis, including rheumatoid arthritis, peridental disease, psoriasis, and smooth muscle cell migration (e.g., restenosis). They have also been found to be effective as antiviral, antifungal and antimicrobial. Thus, compounds that selectively inhibit or antagonize α _ν β3 would be effective in the treatment of these conditions.

It was demonstrated that integrins _α ν β3θ other containing integrins _and are believed to bind to macromolecules matrix containing the Arg-Gly-Asp (RGD). Compounds containing sequences

RGDs resemble extracellular matrix ligands by binding to cell surface receptors. However, it is also known that RGD peptides are generally non-selective for RGD-dependent integrins. For example, most RGD peptides that bind to α _ν β ₃ also bind to α _ν β ₅ , α _ν βι, and an _b β3. Antagonic effect of platelet and _{t b} β3 (also known as fibrinogen receptor) is characterized by blocking platelet aggregation in humans. In order to eliminate the side effect of bleeding in the treatment of these conditions or diseases associated with integrin α _ν β3, it would be beneficial to develop compounds that are selective α _ν β ₃ antagonists with the opposite effect as α _t | _b β ₃ .

Invasion of tumor cells takes place in three steps: 1) attaching the tumor cell to the extracellular matrix; 2) proteolytic dissolution of the matrix; and 3) cell movement across the dissolved barrier. The process can proceed rapidly to produce metastases at sites different from the original tumor.

Seftor et al. (Proc. Natl. Acad. Sci. USA, Vol. 89 (1992) 1557-1561) found that integrins α _ν β ₃ have biological function in cellular invasion of melanoma. Montgomery et al., (Proc. Natl. Acad. Sci. USA, Vol. 91 (1994) 8856-60) has shown that integrin α _ν β ₃ expressed on human melanoma cells induces a survival signal protecting cells from apoptosis. The benefit would be to mediate the pathway of tumor cell metastases by interfering with the α _ν β ₃ integrin receptor for cell adhesion to prevent tumor metastasis.

Brooks et al. (Cell, Vol. 79 (1994) 1157-1164) has shown that α _ν β ₃ antagonists have a therapeutic effect in the treatment of neoplasia (inhibition of solid tumor growth), since systemic administration of α _ν β ₃ antagonists causes marked regression of various histologically distinct human tumors. .

Receptor integrin α _ν β ₃ adhesion has been established as an indicator of angiogenic blood vessels in chickens and humans, and therefore this receptor plays an important role in angiogenesis or neovascularization. Angiogenesis is characterized by invasion, migration and proliferation of smooth muscle cells and endothelial cells. Α _ν β ₃ antagonists selectively inhibit this process by stimulating cell apoptosis in the neovasculature. The growth of new blood vessels or angiogenesis also contributes to a pathological condition, such as diabetic retinopathy including macular degeneration (Adamis et al., Amer. J.Ophtal., Vol. 118, (1994) 445-450) and rheumatoid arthritis (Peacock et al., J. Exp. Med., Vol. 175, (1992), 1135-1138). Anta 9 9 9 9 9 9 9 9 9 9999 99 9 gonists α _ν β ₃ would therefore be therapeutically useful substances in the treatment of these conditions associated with neovascularization (Brooks et al., Science, Vol. 264, (1994), 569-571).

The cell surface receptor α _ν β ₃ has been reported to be the major integrin on osteoclasts responsible for bone attachment. Osteoclasts cause cellular resorption, and if this bone resorption activity predominates over bone formation activity, the result is osteoporosis (loss of bone mass), leading to increased bone fracture, disability and death. Α _ν β ₃ antagonists have been shown to have significant ability to inhibit osteoclastic activity in vitro [Sato et al., J. Cell. Biol., Vol. 111 (1990) 1713-1723] and in vivo [Fisher et al., Endocrinology, Vol. 132 (1993) 1411-1413], Antagonism α _ν β ₃ reduces bone resorption and therefore maintains a normal balance between bone formation and bone loss. It would be useful to create ostelastic antagonists α _ν β ₃ . which are potent inhibitors of bone resorption and are thus useful in the treatment or prevention of osteoporosis.

The role of integrin α _ν β ₃ in smooth muscle cell migration is also a therapeutic goal in preventing or inhibiting neointimal hyperplasia, which is a major cause of restenosis following vascular interventions (Choi et al., J. Vasc. Surg. Vol. 19 (1) (1994) ) 12534). The benefit would be the prevention or inhibition of neointimal hyperplasia by pharmaceutical agents to prevent or inhibit restenosis.

White (Current Biology, Vol. 3 (9) (1993) 596-599) reported that adenovirus uses α _ν β ₃ to enter the host cell. Integrin appears to be essential for endocytosis of the viral particle and may be indispensable for penetration of the viral genome into the cytoplasm of the host cell. Compounds that inhibit α _ν β ₃ would therefore find utility as antiviral agents.

SUMMARY OF THE INVENTION

The invention provides classes of compounds of Formula I

or a pharmaceutically acceptable salt thereof, wherein

A ^ ^x kA! HET:

of ¹ is a 5 to 8 membered monocyclic heterocyclic ring, optionally unsaturated, containing 1 to 4 heteroatoms, selected from the group consisting of O, N or S, wherein X ¹ is selected from the group consisting of CH, CH ₂ , N, NH , O and S;

yi _n ^ NR ⁷ wherein Y ¹ is selected from the group consisting of NR ^2, O and S;

R ² is selected from the group consisting of H; alkyl; aryl; hydroxy; alkoxy; cyano;

nitro; amino; alkenyl; alkynyl; amido; alkylcarbonyl; arylcarbonyl; alkoxycarbonyl; aryloxycarbonyl; haloalkylcarbonyl; haloalkoxycarbonyl; alkylthiocarbonyl; arylthiocarbonyl; acyloxymethoxycarbonyl; alkyl optionally substituted with one or more substituents selected from lower alkyl, halogen, hydroxy, haloalkyl, cyano, nitro, carboxyl, amino, alkoxy, aryl or aryl optionally substituted with one or more substituents selected from halo, haloalkyl, lower alkyl, alkoxy, φ · · · · · · • · · · φ · φ · • · • · φ · φ · φ · φ · · · Cyano, alkylsulfonyl, alkylthio, nitro, carboxyl, amino, hydroxyl, sulfuric acid, sulfonamide, aryl, fused aryl, monocyclic heterocycles or fused monocyclic heterocycles; aryl optionally substituted with one or more substituents selected from halogen, haloalkyl, hydroxy, lower alkyl, alkoxy, methylenedioxy, ethylenedioxy, cyano, nitro, alkylthio, alkylsulfonyl, sulfuric acid, sulfonamide, carboxyl derivatives, amino, aryl, fused aryl, monocyclic heterocycles or fused monocyclic heterocycles; and monocyclic heterocycles which are optionally substituted with one or more substituents selected from halogen, haloalkyl, lower alkyl, alkoxy, amino, nitro, hydroxy, carboxy, carboxyl derivatives, cyano, alkylthio, alkylsulfonyl, sulfuric acid, sulfonamide, aryl or fused aryl; or

R ² together with R ⁷ forms a 4 to 12 membered heterocycle containing two nitrogen atoms optionally substituted by one or more substituents selected from the group consisting of lower alkyl, thioalkyl, alkylamino, hydroxy, keto, alkoxy, halogen, phenyl, amino, carboxyl or carboxyl ester; fused phenyl;

R ² together with R ⁷ forms a 4 to 12 membered heterocycle containing one or more heteroatoms selected from the group consisting of O, N and S, optionally unsaturated;

R ² together with R ⁷ forms a 5- to 9-membered heteroaromatic ring optionally substituted with one or more substituents selected from the group consisting of lower alkyl, phenyl, alkoxy and hydroxy;

R ² together with R ⁷ forms a 5 membered heteroaromatic ring fused to an aryl or heteroaryl ring;

R ⁷ (unless present with R ² ) and R ⁸ are independently selected from the group consisting of H; alkyl; alkenyl; alkynyl; aralkyl; amino; alkylamino; hydroxy; alkoxy; arylamino; amido, alkylcarbonyl, arylcarbonyl; alkoxycarbonyl; aryloxy; aryloxycarbonyl; haloalkylcarbonyl; haloalkoxycarbonyl; alkylthiocarbonyl; arylthiocarbonyl, acyloxymethoxycarbonyl; cycloalkyl; bicycloalkyl; aryl; acyl; benzoyl; alkyl optionally substituted with one or more substituents selected from the group consisting of lower alkyl, halogen, hydroxy, haloalkyl, cyano, nitro, carboxyl derivatives, amino, alkoxy, thio, alkylthio, sulfonyl, aryl, aralkyl, aryl optionally substituted with one ···· 4 or more substituents selected from the group consisting of halogen , haloalkyl, lower alkyl, alkoxy, methylenedioxy, ethylenedioxy, alkylthio, haloalkylthio, thio, hydroxy, cyano, nitro, carboxyl derivatives, aryloxy, amido, acylamino, amino, alkylamino, dialkylamino, trifluoroalkoxy, trifluoromethyl, sulfonyl, alkylsulfonyl, haloalkylsulfonyl, sulfuric acid, sulfonamide, aryl, fused aryl, monocyclic heterocycles, fused monocyclic heterocycles; aryl optionally substituted with one or more substituents selected from the group consisting of halide, haloalkyl, lower alkyl, alkoxy, methylenedioxy, ethylenedioxy, alkylthio, haloalkylthio, thio, hydroxy, cyano, nitro, carboxyl derivatives, aryloxy, amido, acylamino, amino, alkylamino, dialkylamino , trifluoroalkoxy, trifluoromethylsulfonyl, alkylsulfonyl, sulfuric acid, sulfonamide, aryl, fused aryl, monocyclic heterocycles or fused monocyclic heterocycles; monocyclic heterocycles optionally substituted with one or more substituents selected from the group consisting of halide, haloalkyl, lower alkyl, alkoxy, aryloxy, amino, nitro, hydroxy, carboxyl derivatives, cyano, alkylthio, alkylsulfonyl, aryl, fused aryl; monocyclic and bicyclic heterocyclic alkyls; -SChR ¹⁰ , wherein R ¹⁰ is selected from the group consisting of alkyl, aryl and monocyclic heterocycles, all optionally substituted with one or more substituents selected from the group consisting of halo, haloalkyl, alkyl, alkoxy, cyano, nitro, amino, acylamino, trifluoroalkyl, amido alkylaminosulfonyl, alkylsulfonyl, alkylsulfonylamino, alkylamino, dialkylamino, trifluoromethylthio, trifluoroalkoxy, trifluoromethylsulfonyl, aryl, aryloxy, thio, alkylthio and monocyclic heterocycles; and

O ti

CR wherein R ¹⁰ is as defined above;

NR ⁷ and R ⁸ together form a 4 to 12 membered monocyclic or bicyclic ring containing one nitrogen atom optionally substituted with one or more substituents selected from the group consisting of lower alkyl, carboxal derivatives, aryl or hydroxy, and wherein said ring optionally contains a heteroatom selected from including O, N and S;

R ⁵ is selected from the group consisting of H, alkyl, alkenyl, alkynyl, benzyl, and phenethyl; or A is

wherein Y ² is selected from the group consisting of alkyl; cycloalkyl; bicycloalkyl; aryl; monocyclic heterocycles; alkyl optionally substituted with aryl, which may also be substituted with one or more substituents selected from the group consisting of halogen, haloalkyl, alkyl, nitro, hydroxy, alkoxy, aryloxy, aryl, or fused aryl; aryl optionally substituted with one or more substituents selected from the group consisting of halogen, haloalkyl, hydroxy, alkoxy, aryloxy, aryl, fused aryl, nitro, methylenedioxy, ethylenedioxy, or alkyl; alkynyl; alkenyl; -SR ⁹ and OR ⁹ , wherein R ⁹ is selected from the group consisting of H; alkyl; aralkyl; aryl; alkenyl; and alkynyl; or R ⁹ together with R ⁷ forms a 4 to 12 membered heterocyclic ring containing one nitrogen atom and one sulfur atom or one oxygen atom, optionally substituted with lower alkyl, hydroxy, keto, phenyl, carboxyl or carboxyl ester and fused phenyl; or R ⁹ together with R ⁷ is thiazole; oxazole; benzoxazole; or benzothiazole; and R ⁵ and R ⁷ are as defined above;

Y ² (when Y ² is carbon) together with R ⁷ form a 4 to 12 membered ring containing one nitrogen atom or two nitrogen atoms, optionally substituted with an alkyl, aryl, keto or hydroxy group;

A is ········

wherein R ² and R ⁷ form a 5 to 8 membered heterocycle containing two nitrogen atoms, optionally substituted with one or more substituents selected from the group consisting of lower alkyl hydroxy, alkoxy, keto, phenyl or carboxyl derivatives; and R ⁸ is selected from the group consisting of alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkylcarbonyl, haloalkoxycarbonyl, alkylthiocarbonyl, arylthiocarbonyl or acyloxymethoxy carbonyl; and

R ⁵ is as defined above

R ² and R ⁷ together form a heteroaromatic ring such as imidazole or pyrimidone;

And is

R ^a

NR ²

-N = C

N-R '

Wherein R ² and R ⁷ form a 5 to 8 membered heterocycle containing two nitrogen atoms, optionally substituted with a hydroxy, keto, phenyl or alkyl group; and

R ⁸ is selected from the group consisting of alkylcarbonyl, acylcarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkylcarbonyl, haloalkoxycarbonyl, alkylthiocarbonyl, arylthiocarbonyl and acyloxymethoxycarbonyl;

Z ¹ is one or more substituents selected from the group consisting of H; hydroxy alkyl; alkoxy; aryloxy; halogen; haloalkyl; haloalkoxy; nitro; amino; alkylamino; acylamino; dialkylamino; cyano; alkylthio; alkylsulfonyl; carboxyl derivatives; tri444 4 44

4 4 4 4 4444 44 4 444 4444444 4 4 4 44 44 4 44 444 haloacetamide; acetamide; acyl; aryl; fused aryl; cycloalkyl; thio; monocyclic heterocycles; fused monocyclic heterocycles; and A, wherein A is as defined above; V is selected from the group consisting of -N- (R ⁶ ) -, wherein R ⁶ is selected from the group consisting of H; lower alkyl; cycloalkyl; aralkyl; aryl; and monocyclic heterocycles; or R ® together with Y forms a 4 to 12 membered ring containing one nitrogen atom;

Y, Y ³ , Z and Z ³ are independently selected from the group consisting of hydrogen; alkyl; and cycloalkyl; or Y and Z together form a cycloalkyl; or Y ³ and Z ³ together form a cycloalkyl; n is an integer of 1, 2 or 3; t is an integer of 0.1 or 2;

p is an integer of 0,1 or 3;

R is XR ³ wherein X is selected from the group consisting of O, S and NR ⁴ wherein R ³ and R ⁴ are independently selected from hydrogen; alkyl; alkenyl; alkynyl; haloalkyl; aryl; arylalkyl; sugars; steroids; polyalkylethers; alkylamido; alkyl- (N, N-dialkylamide); pivaloyloxymethyl; and in the case of the free acid, all pharmaceutically acceptable salts thereof;

R ¹ is selected from the group consisting of hydrogen, alkyl, alkenyl; alkynyl; aryl; carboxyl derivatives; haloalkyl; cycloalkyl; monocyclic heterocycles; monocyclic heterocycles optionally substituted with alkyl, halide, haloalkyl, cyano, hydroxy, aryl, fused aryl, nitro, alkoxy, aryloxy, alkylsulfonyl, arylsufonyl, sulfonamide, thio, alkylthio, carboxyl derivatives, amino and amido groups;

alkyl optionally substituted with one or more halo, haloalkyl, hydroxy, alkoxy, aryloxy, thio, alkylthio, alkynyl, alkenyl, alkyl, arylthio, alkylsulfoxide, alkylsulfonyl, arylsulfoxide, arylsulfonyl, cyano, nitro, amino, alkylamino, dialkylamino, alkylsulfonamide, aryls acylamide, carboxyl derivatives, sulfonamide, sulfuric acid, phosphoric acid derivatives, phosphorous acid derivatives, aryl, arylthio, arylsulfoxide, or arylsulfone, in each case optionally substituted on the aryl ring with halogen, alkyl, haloalkyl, cyano, nitro, hydroxy, carboxyl derivatives, alkoxy, aryloxy, amino, alkylamino, dialkylamino, amido, aryl, fused aryl, monocyclic heterocycles; and fused monocyclic heterocycles, monocyclic heterocyclothio, monocyclic heterocyclosulfoxide; and a monocyclic heterocyclosulfone, which may optionally be substituted by a halogen group, a haloalkyl, nitro, hydroxy, alkoxy, a fused aryl or an alkyl group;

alkylcarbonyl, haloalkylcarbonyl and arylcarbonyl;

aryl optionally substituted at one or more positions by halogen, haloalkyl, alkyl, alkoxy, aryloxy, methylenedioxy, ethylenedioxy, alkylthio, haloalkylthio, thio, hydroxy, cyano, nitro, acyloxy, carboxy derivatives, carboxyalkoxy, amido, acylamino, amino, alkylamino dialkylamino, trifluoroalkoxy, trifluoromethylsulfonyl, alkylsulfonyl, sulfuric acid, sulfonamide, aryl, fused aryl, monocyclic heterocycles and fused monocyclic heterocycles; and

OR ⁷ ii from —C — N ^X R® wherein R ⁷ and R ⁸ are as defined above and together with the nitrogen R ⁷ and R ⁸ comprise an amino acid and R ¹¹ is selected from the group consisting of H, alkyl, aralkyl, alkenyl, alkynyl, haloalkyl or haloalkynyl or R ¹¹ together with Y form a 4 to 12 membered ring containing one nitrogen atom.

Another object of the invention is a pharmaceutical composition comprising a compound of formula I. Such compounds and compositions are useful as selective inhibitors and integrin antagonists _and β _3, and therefore in another embodiment the invention provides a method of selectively inhibiting or antagonizing the integrin _α ν β _third The invention further encompasses the treatment or inhibition of pathological conditions associated with, for example, osteoporosis, humoral hypercalcemia in malignancy, Paget's tumor, tumor metastasis, solid tumor growth (neoplasia), angiogenesis including tumor angiogenesis, retinopathy including macular degeneration and diabetic tetinopathy, arthritis including arthritis, peridental disease, psoriasis, smooth muscle cell migration, and restenosis in a mammal in need of such treatment. Moreover, these pharmaceutical agents are useful as antiviral and antimicrobial agents.

The invention provides a class of compounds of formula I as described above.

A preferred embodiment of the invention is a compound of formula II

(II) wherein R ³² is H, alkyl, alkoxyalkyl, (II), or (II), wherein R ³² is H, alkyl, alkoxyalkyl, aminoalkyl, dialkylaminoalkyl, wherein the alkyl group is optionally substituted with one or more substituents selected from the group consisting of hydroxy, alkoxy, amino, alkylamino, dialkylamino, aryl- or alkylsulfonyl, carboxyl, and carboxyl derivatives, and other variations as described in the general formula AND.

Another preferred embodiment of the invention is a compound of formula (II): A compound of formula (II). ·· · · · · · · · · · · · ·

is optionally substituted, other variations being defined by Formula I above.

Another preferred embodiment of the invention is a compound of formula II

where ········ · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

Yippee

optionally substituted and other variations are defined by the general formula I above.

Another preferred embodiment of the invention is a compound of formula II

Λ.

(II), "A ¹¹ !" Het!

'* t

J ^e NM> 32

N-N /

optionally substituted and other variations are defined by Formula I above.

Another preferred embodiment of the invention is a compound of formula II wherein

···· this • to it «·

optionally substituted and other variations are defined in Formula I above.

The invention further provides pharmaceutical compositions comprising therapeutically effective amounts of compounds of formulas I or II.

The invention also provides a method for the selective inhibition or antagonistic effect _of α _v β ₃ integrin and more particularly relates to a method of inhibiting bone resorption, peridental disease, osteoporosis, humoral hypercalcemia in malignant disease, Paget's tumor, tumor metasases, solid tumor growth (neoplasia), angiogenesis, including tumor angiogenesis, retinopathy, including macular degeneration and diabetic retinopathy, arthritis, including rheumatoid arthritis, peridental disease, psoriasis, smooth muscle cell migration, and restenosis, by administering a therapeutically effective amount of a compound of Formulas I to VI together with a pharmaceutically acceptable carrier.

The following is a list of definitions of the different terms used here:

The term "alkyl" or "lower alkyl" as used herein refers to an unbranched or branched chain of hydrocarbon radicals containing 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms.

Examples of such radicals are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, neopentyl, hexyl, isohexyl and the like.

The term "alkenyl" or "lower alkenyl" refers to unsaturated acyclic hydrocarbon radicals containing at least one double bond and 2 to 6 carbon atoms, wherein the carbon double bond may be in the cis or trans position of the alkenyl group relative to the groups substituted on the carbon atoms with double bond. Examples of such groups are ethenyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl and the like.

· · € * © © © © © © © © © © © © · · * * · · · * · * * * * * * 9 ·· ·· ©

The term "alkynyl" or "lower alkynyl" as used herein refers to non-cyclic hydrocarbon radicals containing one or more triple bonds and 2 to 6 carbon atoms. Examples of such groups are ethynyl, propynyl, butynyl, pentynyl, hexynyl and the like.

The term "cycloalkyl," as used herein, represents saturated or partially unsaturated cyclic carbon radicals containing 3 to 8 carbon atoms, and more preferably 4 to 6 carbon atoms. Examples of such cycloalkyl radicals include cyclopropyl, cyclopropenyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-cyclohexen-1-yl and the like.

The term "aryl" as used herein refers to aromatic cyclic systems containing one or more aromatic rings. The term includes aromatic radicals such as phenyl, pyridyl, naphthyl, thiophene, furan, biphenyl and the like.

The term "cyan" as used herein characterizes the radical of the formula

CN.

As used herein, the terms "hydroxy and" hydroxyl "are synonyms and are characterized by the radical of formula --OH.

The radical of the formula refers to a 5 to 10 membered heterocycle, optionally unsaturated, containing 1 to 3 heteroatoms selected from the group consisting of O, N and S; wherein X ¹ is CH, N, O or S. Characteristic examples of such radicals include pyridones, pyridines, pyrimidines, imidazoles, oxazoles, isoxazoles, thiazoles, pyridazines, thiophenes, furans, pyrazoles and bicyclic heterocycles such as benzimidazole, imidazopyridine, benzofuran and the like. .

The term "lower alkylene" or "alkylene" as used herein refers to divalent linear or branched saturated hydrocarbon radicals containing 1 to 6 carbon atoms.

·········· · ·

9 fcfcfc ···· fcfc · «fcfc fcfc • fc« 4 * • · fc • · (fc fcfcfc f fcfcfc • fcfcfcfc «• ··« • fcfc fcfcfc

The term "alkoxy" as used herein refers to an unbranched or branched chain containing oxygen radicals of the formula -OR ²⁰ , wherein R ²⁰ is an alkyl group as defined above. Examples of alkoxy groups include methoxy, ethoxy, n-propoxy, n-butoxy, isopropoxy, isobutoxy, sec-butoxy, t-butoxy and the like.

The term "arylalkyl or" aralkyl "refers to a radical of the formula

R ²² - R ²¹ wherein R ²¹ is aryl as defined above and R ²² is alkylene as defined above. Examples of alkyl groups include benzyl, pyridylmethyl, naphthylpropyl, phenethyl and the like.

The term "nitro" as used herein represents a radical of the formula j-NO 3.

The term "halo" or "halogen" represents bromo, chloro, fluoro and iodo.

As used herein, the term "haloalkyl" refers to alkyl groups as defined above substituted with one or more identical or different halogen groups on one or more carbon atoms. Examples of haloalkyl groups include trifluoromethyl, dichloroethyl, fluoropropyl and the like.

The term "carboxyl" or "carboxy" as used herein refers to a radical of the formula -COOH.

The term "carboxyl ester" refers to a radical of the formula -COOR ²³ wherein R ²³ is selected from the group consisting of H, alkyl, aralkyl or aryl as defined above.

The term "carboxyl derivative" as used herein refers to a radical of the general formula

Y ⁶ C-Y-R ²³ wherein Y ⁶ and Y ⁷ are independently selected from the group consisting of O, N or S and R ²³ is selected from the group consisting of H, alkyl, aralkyl or aryl as defined above.

The term "amino" as used herein represents a radical of the formula -NH ₂ .

The term "alkylsulfonyl" or "alkylsulfone" as used herein refers to a radical of the formula:

O

wherein R ²⁴ is alkyl as defined above.

The term "alkylthio" refers to a radical of the formula -SR ²⁴ , wherein R ²⁴ is alkyl as defined above. As used herein, the term "sulfuric acid" refers to a radical of the formula

O

wherein R ²⁵ is alkyl as defined above.

The term "sulfonamide" refers to a radical of the formula

wherein R ⁷ and R ⁸ are as defined above.

The term "fused aryl," as used herein, represents an aromatic ring wherein the aryl groups as defined above are combined into one or more phenyl rings. The term "fused aryl" is a naphthyl radical and the like.

The term "monocyclic heterocycle" or "monocyclic heterocyclic" as used herein refers to a monocyclic ring containing 4 to 12 atoms, more preferably 5 to 10 atoms, of which 1 to 3 atoms are heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur thereto that in the case of two or more different heteroatoms one of the heteroatoms must be nitrogen. A characteristic example of such a monocyclic heterocycle is imidazole, furan, pyridine, oxazole, pyran, triazole, thiophene, pyrazole, thiazole, thiadiazole and the like.

The term "fused monocyclic heterocycle" as used herein refers to a monocyclic heterocycle, as defined above, fused with benzene. Examples of these fused monocyclic heterocycles include benzofuran, benzopyran, benzodioxole, benzothiazole, benzothiophene, benzimidazole. etc.

The term "methylenedioxy," as used herein, refers to the radical ti · · · · · · · · · ··· ·····

H ° t-° and the term "ethylenedioxy" refers to a radical

The term "4-12 membered heterocycle containing two nitrogen atoms," as used herein, refers to a radical of the formula

wherein m is 1 or 2 and R ¹⁹ is H, alkyl, aryl or aralkyl and more preferably relates to a 4 to 9 membered ring and includes, for example, imidazoline.

The term "5-membered optionally substituted heteroaromatic ring" as used herein includes, for example, a radical of the formula

and "5-membered heteroaromatic ring fused to phenyl" refers to this "5-membered heteroaromatic ring" with phenyl attached thereto.

A characteristic example of such a 5-membered heteroaromatic ring fused to phenyl is benzimidazole.

The term "bicycloalkyl," as used herein, refers to a bicyclic hydrocarbon radical containing from about 6 to 12 carbon atoms that is saturated or partially unsaturated.

The term "acyl" as used herein refers to a radical of the formula

Wherein R ²⁶ is alkyl, alkenyl, alkynyl, aryl or aralkyl and is optionally substituted as defined above. This radical includes acetyl, benzoyl and the like groups.

As used herein, the term "thio" refers to a radical of the formula

SH.

The term "sulfonyl" as used herein refers to a radical of the formula

II

Wherein R ²⁷ is alkyl, aryl or aralkyl as defined above.

The term "haloalkylthio," as used herein, refers to a radical of the formula

-SR ²⁸ wherein R ²⁸ is haloalkyl as defined above.

The term "aryloxy" as used herein refers to a radical of the formula wherein R ²⁹ is aryl as defined above.

The term "acylamine," as used herein, refers to a radical of the formula: 9 9 9 9 9 9 9 9 9 9 9 9 9 9

9 9 9 9 9999999 9

99 99 99 99 999

O r »_ £ _nh_ | wherein R ³⁰ is alkyl, aralkyl or aryl as defined above.

The term "amido" as used herein refers to a radical of the formula

O li

NH2.

The term "alkylamino" as used herein refers to a radical of the formula -NHR ³² wherein R ³² is alkyl as defined above.

The term "dialkylamino" as used herein refers to a radical of the formula -NR ³³ R ³⁴ , wherein R ³³ and R ³⁴ are the same or different alkyl groups as defined above.

The term "trifluoromethyl" as used herein refers to a radical of the formula

CF,.

The term "trifluoroalkoxy" as used herein refers to a radical of the formula

Wherein R ³⁵ , wherein R ³⁵ is a bond or alkylene, as defined above.

• ·

The term "alkylaminosulfonyl," as used herein, refers to a radical of the formula

O

R ³ ® — NS — i H u ¹

O wherein R ³⁶ is alkyl as defined above.

The term "alkylsulfonylamino" refers to a radical of the formula

R ³⁶ —S-NH — 1 li ·

O wherein R ³⁶ is alkyl as defined above.

The term "trifluoromethylthio" as used herein refers to a radical of the formula

F3C — S— | ·

The term "trifluoromethylsulfonyl" as used herein refers to a radical of the formula

O ¹¹ l

F3C '-S — I 11 *

O

The term "4-12 membered nitrogen-containing cyclic or bicyclic ring" refers to a saturated or partially unsaturated monocyclic or bicyclic ring of 4 to 12 atoms, more preferably a ring of 4 to 9 atoms wherein one of the atoms is nitrogen. These rings may additionally optionally contain heteroatoms selected from the group consisting of nitrogen, oxygen or sulfur. This group includes morpholine, piperidine, piperazine, thiomorpholine, pyrrolidine, proline, azacycloheptene and the like.

The term "benzyl" as used herein refers to the radical fefe fefe fefe fefe feef fe fe fe fe fe ··· · · · ··············· · fefe · fefe ··· h ^ch '-Q

The term "phenethyl" as used herein refers to the radical | —CH2CH2—

The term "4-12 membered heterocyclic ring containing one nitrogen, sulfur and oxygen atom" as used herein refers to a ring containing 4 to 12 atoms, more preferably 4 to 9 atoms, wherein at least one atom is nitrogen and at least one atom is oxygen or sulfur. This definition includes rings such as thiazoline and the like.

The term "arylsulfonyl" or "arylsulfone" as used herein refers to a radical of the formula

O

R ³⁷ —S — l ii *

Wherein R ³⁷ is aryl as defined above.

The term "alkylsulfoxide" or "arylsulfoxide" as used herein refers to radicals of the formula

wherein R ³⁸ is alkyl or aryl as defined above.

As used herein, the term "phosphorous acid derivative" refers to a radical of the formula · «· · φ · · · · · · · · fosfor fosfor fosfor fosfor fosfor fosfor fosfor fosfor

Ο

1- p -OR ³⁹ OR ⁴⁰ wherein R ³⁹ and R ⁴⁰ may be H, alkyl, aryl or aralkyl.

The term "phosphoric acid derivatives," as used herein, refers to a radical of the formula

O-P-OR ⁴¹ H wherein R ⁴¹ is H, alkyl, aryl or arylalkyl as defined above.

The term "arylthio," as used herein, refers to a radical of the formula

SR ⁴² wherein R ⁴² is aryl as defined above.

The term "monocyclic thioheterocycle" as used herein refers to a radical of the formula | -SR ⁴³ wherein R ⁴³ is a monocyclic heterocycle radical as defined above.

The term "monocyclic heterocycle sulfoxide" and "monocyclic heterocycle sulfone" refers to a radical or formula oo LSR ⁴³ "F $ -R ⁴³ o

wherein R ⁴³ is a monocyclic heterocycle radical as defined above.

The term "alkylcarbonyl," as used herein, refers to a radical of the formula

wherein R ⁵⁰ is alkyl as defined above.

The term "arylcarbonyl," as used herein, refers to a radical of the formula

O

wherein R ⁵¹ is aryl as defined above.

The term "alkoxycarbonyl," as used herein, refers to a radical of the formula

wherein R ⁵² is an alkoxy group as defined above.

The term "aryloxycarbonyl," as used herein, refers to a radical of formula o

Wherein R ⁵¹ is aryl as defined above.

The term "haloalkylcarbonyl," as used herein, refers to a radical of the formula

wherein R ⁵³ is haloalkyl as defined above.

The term "haloalkoxycarbonyl," as used herein, refers to a radical of the formula

O

R ⁵³ —OC—

Wherein R ⁵³ is haloalkyl as defined above.

The term "alkylthiocarbonyl" refers to a radical of the formula

The r-C_ _S ⁵⁰ wherein R ⁵⁰ is alkyl as defined above.

The term "arylthiocarbonyl," as used herein, refers to a radical of the formula

O

R ⁵¹ S-C_ wherein R ⁵¹ is aryl as defined above.

The term "acyloxymethoxycarbonyl," as used herein, refers to a radical of the formula

O

AND"

R ⁵⁴ -O-CH ₂ -O_C- wherein R ⁵⁴ is acyl as defined above.

The term "arylamine" as used herein refers to a radical of the formula R ^{S 1} -NH- wherein R ⁵¹ is aryl as defined above.

The term "polyalkyl ether" as used herein refers to commonly used glycols such as triethylene glycol, tetraethylene glycol, polyethylene glycol and the like.

The term "alkylamide" refers to a radical of formula o

>>

R -NH-C- wherein R ⁵⁰ is alkyl as defined above.

The term "Ν, Ν-dialkylamide" refers to a radical of the formula

O

II

Where R ⁵⁰ is the same or different alkyl group as defined above.

The term "pivaloyloxymethyl" as used herein refers to a radical of the formula

Me

O

II 'C — C — O — CH2

Me

The term "acyloxy" refers to a radical of the formula R 6 -O-, wherein R ⁵⁵ is acyl as defined above.

As used herein, the term "composition" refers to a product that results from a mixture or combination of more than one component or ingredient.

The term "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable substance, composition or carrier, for example, a liquid or solid filler, diluent, carrier, diluent or encapsulating agent as carrier or carrier of a chemical.

The term "therapeutically effective amount" means that amount of a drug or pharmaceutical agent that elicits a biological or clinical response to a tissue, system or animal selected by a researcher or physician.

The following is a list of abbreviations and corresponding explanations as used interchangeably here:

H ¹ -NMR = proton nuclear magnetic resonance

AcOH = acetic acid

Ar = argon

BH3-THF = borane-tetrahydrofuran complex • · • ·

9

999 9 9 · 9 · 9 fi 9 fi 99 9 9 9 9 99999

9 9

9

Βη = benzyl

BOC = t-butoxycarbonyl

But Li = butyllithium

Cat. = catalytic amount

CDMT = 2-chloro-4,6-dimethoxytriazine

CH ₂ Cl ₂ = dichloromethane

CH 3 CN = acetonitrile

CH ₃ I = iodomethane

CHN analysis = elemental carbon / hydrogen / nitrogen analysis

CHNCI analysis = elemental analysis carbon / hydrogen / nitrogen / chlorine CHNS analysis = elemental analysis carbon / hydrogen / nitrogen / sulfur DAST = diethylaminosulfurtrifluoride

DCC = 1,3-dicyclohexylcarbodiimide

DCM = dichloromethane

DIBAL = diisobutylaluminum hydride

DIEA = diisopropylethylamine

D1 water = deionized water

DMA = N, N-dimethylacetamide

DMAC = Ν, Ν-dimethylacetamide

DMAP = 4- (N, N-dimethylamino) pyridine

DMF = N, N-dimethylformamide

DSC = disuccinyl carbonate

EDCI = 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride Et = ethyl

Et ₂ O = diethyl ether

Et ₃ N = triethylamine

EtOAc = ethyl acetate

EtOH = ethanol

FAB MS = fast atom bombardment mass spectrometry g = gram (grams) ································ ··

GIHA = meta-guanidinohippuric acid

GIHA HCl ~ meta-guanidinhippuric acid hydrochloride

Gly = glycine

HMPA = hexamethylphosphoramide

HOBT = 1-hydroxybenzotriazole hydrate

HPLC = high performance liquid chromatography

IBCF = isobutyl chloroformate i-Pr = isopropyl i-Prop = isopropyl

K2CO3 = potassium carbonate

KMnO ₄ = potassium permanganate

KOH = potassium hydroxide

KSCN = potassium thiocyanate

L = liter

LiOH = lithium hydroxide

MCPBA = m-chloroperbenzoic acid or m-chloroperbenzoic acid Me = methyl Mel = methyl iodide MeOH = methanol

MEMCI = methoxyethoxymethyl chloride

MesCl = methanesulfonyl chloride mg = milligram

MgSO ₄ = magnesium sulfate ml = milliliter mL = milliliter

MS = mass spectrometry

MTBE = methyl t-butyl ether

N ₂ = nitrogen

NaCNBH ₃ = sodium cyanoborohydride

NaH = sodium hydride

NaHCO ₃ = sodium bicarbonate

NaOH = sodium hydroxide

NaOMe = sodium methoxide

Na ₂ PO ₄ = sodium phosphate

Na ₂ SO 4 = sodium sulfate

NEt ₃ = triethylamine

NH ₄ HCO ₃ = ammonium bicarbonate

NH ₄ ⁺ HCO ₂ '= ammonium formate

NH ₄ OH = ammonium hydroxide

NMM = N-methylmorpholine

NMP = 1-methyl-2-pyrrolidinone

NMR = nuclear magnetic resonance

Pd / C = palladium on carbon

Ph = phenyl

Pt / C = platinum on carbon

RPHPLC = reverse phase HPLC

RT = room temperature t-BOC = t-butoxycarbonyl TFA = trifluoroacetic acid THF = tetrahydrofuran TLC thin-layer chromatography TMEDA = tetramethylethylenediamine TMS = trimethylsilyl Δ = heating of the reaction mixture

The compounds of formulas (I) to (VI) may exist in various isomeric forms and include all isomeric forms. Tautomeric forms are also included, as are pharmaceutically acceptable salts of these isomers and tautomers.

In structures and formulas, the bond through the ring boundary may belong to any atom of the ring.

• · · · · · · · · · · · ·

The term "pharmaceutically acceptable salt" refers to a salt prepared by reacting a compound of Formula I with an acid, the anion of which is generally considered to be ingested by humans. Examples of pharmaceutically acceptable salts include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfate, phosphate, acetate, propionate, lactate, maleate, malate, succinate, quaternary salts and the like. All pharmaceutically acceptable salts may be prepared in conventional manner. (See Berge et al., J. Pharm. Sci., 66 (1), 1-19 (1977) for further examples of pharmaceutically acceptable salts.)

For selective inhibition or antagonistic effect and _in β3 integrins, the compounds of the invention may be administered orally, parenterally or by inhalation of a spray or locally in unit amounts of formulations containing conventional pharmaceutically acceptable carriers, adjuvants, binders. The term parenterally as used herein includes, for example, subcutaneous, intravenous, intramuscular, intrastemal and infusion techniques or intraperitoneal.

The compounds of the invention are administered by any suitable route in the form of a pharmaceutical composition adapted for the method and in an amount effective for the treatment. Therapeutically effective amounts of the compounds prevent or suspend the process or treat the clinical condition readily as determined by one skilled in the art using preclinical and clinical approaches known in the medical arts.

The invention thus provides a method for treating conditions mediated by selectively inhibiting or antagonním effect _α ν β3 cell surface receptor; the method comprising administering a therapeutically effective amount of a compound selected from the class of compounds described by Formulas I to VI wherein one or more compounds of Formulas I to VI are administered in conjunction with one or more non-toxic, pharmaceutically acceptable carrier and / or adjuvant (collectively referred to herein as "Carriers") and optionally with other active ingredients. More specifically, the invention relates to a method of inhibiting α _ν β ₃ receptor surface cells. More preferably, the invention relates to a method of inhibiting bone resorption, treating osteoporosis, inhibiting humoral hypercalcemia in malignancy, treating Paget's tumor, inhibiting tumor metastasis, inhibiting neoplasia (solid tumor growth), inhibiting angiogenesis including angiogenesis, treating retinopathy including macular degeneration and diabetic retinopathy , inhibiting arthritis, psoriasis and peridental disease, and inhibiting smooth muscle cell migration including restenosis.

444445 4 4 4 4 4 4

4 4 4 4 4

4 4 · · · · · · · · · · · · · · · 4 4 ·

4 4 4 · · 4 · · ·

4 · · · · 4 4 4 44

Based on standard laboratory experimental techniques and procedures generally known and validated by those skilled in the art, as well as when compared to compounds of known use, the compounds of formula I can be used in the treatment of patients suffering from the above pathological conditions. One of ordinary skill in the art will recognize that the choice of the most suitable compound of the invention will depend on the ability of one of ordinary skill in the art and will depend on many factors including estimation of results obtained in standard measurements and animal models.

Treatment of a patient suffering from one of the pathological conditions comprises administering an amount of a compound of Formula I that is therapeutically effective to control the condition or prolong the life of the patient than is expected without such treatment. As used herein, the term "inhibition" of a condition means slowing, interrupting, stopping or disappearing a condition and does not necessarily mean total exclusion of the condition. It is believed that prolonging the patient's life beyond the significantly beneficial effect as such also means that the condition control is beneficial to some extent.

As mentioned above, the compounds of the invention can be used in a variety of biological, prophylactic or therapeutic areas. It is contemplated that these compounds are useful in the prevention or treatment of any disease or condition in which integrin α _ν β ₃ plays a role.

The dosage of the compounds of the and / or compositions comprising these compounds is based on a number of factors including the type, age, weight, sex and medical condition of the patient; the severity of the condition; method of application; and activity of the compound. Therefore, the dosage regimen may vary widely. Dosages in an amount of from 0.01 mg to about 1000 mg per kilogram of body weight per day are useful in the treatment of the above conditions.

The active ingredient injected is formulated as a composition wherein, for example, saline, dextrose or water can be used as a suitable carrier. Suitable daily dosages may conveniently be in the range of 0.01 to 10 mg / kg body weight daily injected in multiple doses depending on the factors mentioned above.

For administration to a mammal in need of such treatment, the compounds in a therapeutically effective amount are conveniently mixed with one or more adjuvants for the particular mode of administration. The compounds may additionally be mixed with lactose, sucrose, starch powder, alkanoic cellulose esters, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium, calcium salts of sulfuric and phosphoric acid, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone and / or polyvinyl alcohol and formulated into tablets or capsules for conventional use. The compounds may optionally be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride and / or various buffers. Other adjuvants and routes of administration are generally known in the pharmaceutical art.

The pharmaceutical compositions of the invention may be used in conventional pharmaceutical procedures, for example sterilization, and / or may contain conventional pharmaceutical adjuvants, for example, preservatives, stabilizers, humectants, emulsifiers, buffers and the like.

General synthetic procedures for the preparation of the compounds useful according to the invention are shown in Schemes 1 to 15. At the same time, explanations, current procedures, and various aspects of the invention are set forth where appropriate. The following schemes and examples are intended to be illustrative of the invention and are not to be construed as limiting the scope and spirit thereof. Those skilled in the art will readily appreciate that known variations of the conditions and procedures described in the Schemes and Examples may be used to synthesize the compounds of the invention.

Unless otherwise stated, all starting materials and equipment used were commercially available.

Scheme 1 'V

Ί07

NH 3 2) hq HCl 1) DIEA

FfeO / Dioxane

Hjl

.1

CpjH

·· 99

Scheme 2

··· 9 · · <

I · ··

9 999

Scheme 3

EtOH / H ₂ O f η NaH, BOC Anhydride

• ••• 44,444 44 • 44 «44,444

444 4444 444 44

4444 44 4 44 4 • 44 44 44 44 444

The first to third diagrams illustrate the preparation method used to prepare the guanidinopyridine / cycloguanidinopyridine carboxylic acid moieties of the invention which are used to attach to the gly-p-amino acid moiety. This can be achieved using other suitable guanidation agents known to those skilled in the art. The procedures in Schemes 1 through 3 can be altered using conventional techniques and methods to prepare similar compounds useful for attachment to the gly-β-amino acid moiety.

Scheme 4 nh p

HjN ^ N ^

H

NH ₂

OEt

1) EtOH Δ

2) NH 2 OH 4 CO ₂ Et =

⁵ γ ^Ν

HN..NH ₂

Y

NH

NaOH

H ₂ O / EtOH

HC!

(A13)

Scheme 4 shows a methodology for preparing a portion of the guanidinothiazolecarboxylic acid of the invention that is used to attach to the gly-β-amino acid portion. The methods in Scheme 4 may be altered using conventional procedures known to those of skill in the art to prepare this and similar compounds useful for attachment to a portion of a glyβ-amino acid.

···· t ·> A A * f ♦ • • « • AA • • AND ♦ · • • • »· • • · A · • A • • • · • • • • AND • • • ·· ·· AND· • • · A ··

Scheme 5

OH

ActOTEtJM

OF

^X TMSjNU / THF

O

X, N N-X- succulent or (Other source * X

OH

EtOH. HCl

H »N

CO £ t

OH • HCl

Y

X • HCl> 1 u ¹

H I *

COjEt

OH

X • · · · · · · · · · · · · · · · · · · ·

Scheme 5 shows a method for preparing a portion of the ethyl- (N-glylamino-3- (3,5-dihalo-2-hydroxyphenylpropionate)) compounds of the invention Briefly, 3,5-halo substituted salicylaldehydes (halo-substituted -2-hydroxybenzaldehydes) were For example, 5-bromosalicylaldehyde is suspended in acetic acid and an equivalent or greater amount of chlorine is added to give 3-chloro-5-bromo-2-hydroxybenzaldehyde.

Some products precipitate and are recovered by filtration. The residue is recovered by diluting the filtrate with water and isolating the precipitate. Mixing the solids and drying gave 3-chloro-5-bromo-2-hydroxybenzaldehyde. 3-Iodo-5-chlorosalicylaldehyde is prepared by reacting 5-chlorosalicylaldehyde with N-iodosuccinimide in DMF and subjecting the reaction mixture to conventional reaction conditions. 3-Iodo-5-bromosalicylaldehyde can be prepared by reacting 5-bromosalicylaldehyde in acetonitrile with potassium iodide and chloramine T. Conventional treatment affords a compound which, upon reaction with hexanes, yields the desired 3-iodo-5-chlorosalicylaldehyde.

Coumarins are readily prepared from salicylaldehyde using a modified Perkin reaction (for example, the Vogel's Textbook of Practical Organic Chemistry, 5th Edition, 1989, page 1040). Halogen-substituted coumarins are converted to 3-aminohydrocoumarins (see JGRico, Tett. Let., 1994, 35, 6599-6602), which are readily opened in an acidic alcohol to give esters of 3-amino-3- (3,5-halogen) -2-hydroxyphenylpropanoic acid.

3-Amino-3- (3,5-halo-2-hydroxy) phenylpropanoic acid esters are converted to N-gly-3-amino-3- (3,5-halo-2-hydroxy) phenylpropanoic acid esters by BocN-gly N-hydroxysuccinimide to form Boc-N-gly-3-amino-3- (3,5-halo-2-hydroxy) phenylpropanoic acid esters, which are converted to the HX salts of N-gly-3-amino-3- (3, 5-halo-2-hadroxylphenylpropanoic acid (wherein X is Cl, Br or I).

Scheme 6

1) NMM DMA (or) DMF (A1-13) -CO 2 H 2 HCl + CIC

2} (CJ + NMM

3) LiOH / 1-fcO

4) TFA

Scheme 6 shows the methodology for coupling the heterocyclic acid moiety (A1-A13) to the gly-p-amine moiety (C) of the invention.

The synthesis of A1-A13 is shown in Schemes 1-4 and synthesis (C) is shown in Scheme 5 (where X and Y are halogens, identical or different).

This procedure can be modified using conventional methods known to those skilled in the art.

Scheme 7 step A

HO ₂ C 1 ^/ CH ₂ CO ₂ H + R ¹ CHO + NH / CH 3 CO ₂

CO2H r ³ oh

-fe ·

HCl gas Δ

© ····· ··· ·· ··· ··· · · · · · · · · ······· · · · · · · · · ··· step Β

BOC — N

CO2H + ocq.

0) / - \

OIM-CHj

DMF (2) (E), NMM in DMF

HCI / J dioxane '->

(F)

Scheme 7 generally shows the preparation of a portion of the gly-β-amino acid molecule (F). The aldehydes (R 'CHO) used in this process are commercially available or can be prepared from commercially available materials using methods known to those skilled in the art.

All other components are commercially available or can be readily synthesized by one skilled in the art.

These methods and conditions can be further modified using conventional techniques to produce similar desired intermediates.

· 0 · · · · · · · · · · · · · · · · · · · · · · · · · ·

0 ·· · · 0 00 0

Scheme 8

O

ΊCN «*>

▼ · ········ · it ··· · · · · · · · · · · · · · · ·

Scheme 8 shows a method useful for attaching a portion of a heterocyclic acid (A1-A13) to a portion of a gly-β-amino acid (F) to form compounds of the invention.

The synthesis of A1-A13 is shown in Schemes 1-4 and synthesis (F) is shown in Scheme 7.

The procedure may be varied using conventional methods known to those skilled in the art. Schemes 9 to 12 show a general process for preparing compounds of the invention.

Scheme 9

<X1

JI merger

<

····················

Scheme 9 shows a general procedure for the synthesis of heterocycles derived from gly-p-amino acid-linked compounds. Reaction of a heterocyclic carboxylic acid with a glyphinoacid under condensation reaction conditions affords intermediate (2). Reduction of the nitro group using catalytic hydrogenation (e.g. Pt / H, H ₂ ) affords the amino intermediate (3). This transformation can also be performed chemically using SnCl ₂ . The amino group may be converted to guanidino or another functional group of formula I using the method described above.

Scheme 10

guanylation

O

Fcfc fcfc fcfc fcfc fcfcfc

Alternatively, the target compound can be prepared by synthesis of the left side of the molecule prior to coupling with the gly-p-amino acid (Scheme 10). The amino group of the amino heterocycle (1) is operably linked to guanidine or other groups (A, Formula I) and then attached to the gly-p-amino acid under standard condensation reaction conditions.

Scheme 11

4444 44 44 4 ·· 4 • 4 · 4 4 4 4 4 44

4 444 4444 444 4

4444 44 4 44

44 44 44 444

Scheme 11 shows the general synthesis of substituted pyridines and pyridine compounds derived from target compounds. Nitration of 6-hydroxynicotinic acid followed by chlorination gives 6-chloro-5-nitronicotinic acid. Reaction of intermediate 3 with a gly-β-amino acid yields product 4. The chloro group in universal intermediate 4 can be easily replaced by different nucleophiles to give product 5. Reduction of nitro group 4 or 5 and further reaction of amino group as outlined in Scheme 9 above, to obtain the target compound.

Scheme 12

Her merging reactions

guanylation o

··············································· ·· · ·· · · ·

Scheme 12 shows the synthesis of substituted pyridine and pyridone compounds derived from the target compounds from the starting 2-amino-6-hydroxypyridine-4-carboxylic acid. The starting material 1 used in the scheme can be prepared by reacting commercially available 2-chloro-6-methoxypyridine-4-carboxylic acid with ammonium hydroxide under high pressure. Reaction of the amino group followed by condensation reaction with gly-3-amino acid as shown in Scheme 10 provides the target compound.

Scheme 13 x

guanylation ¹¹ o:

ď o

· Φ φ φ φ φ • φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ · · ·· ·· ·

Scheme 13 shows the synthesis of isomeric pyridines and pyridones from 6-amino-4-methoxy-picolinic acid starting material. The starting material 1 used in Scheme 13 can be prepared as described in the literature (J. Am. Chem. Soc., 78, 4130, 1956). Functionalization of the amino group in compound 1 by condensation reaction and hydrolysis (as in Scheme 10) yields the target compound.

Scheme 14

nitration guanylation guanylation

to ··· this · ♦ ····· · · · · · · ······ · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ··

Isomeric pyridine and pyridone compounds can be prepared using the procedure of Scheme 14. Key Intermediate 4 can be prepared from 6-chloro-picolinic acid starting material. Oxidation followed by nitration yields the 4-nitropyridine derivative (3). Deoxygenation of the N-oxide, reduction of the nitro group and nucleophilic removal of the chloro group yields intermediate 4. The procedure outlined in the scheme can be used to synthesize the target compounds.

Scheme 15 x

guanylation guanylation

O

O

£ fcfcfcfc fc · fc * · fcfc «fc ··» · · «fcfcfc ··> fcfcfcfc 9 9 9 • fc fcfcfc fcfcfcfc ··· · · ······· fcfcfc

The procedure outlined in Scheme 10 may also be used to synthesize pyrimidine derivatives derived from a target compound (Scheme 15). Isomeric pyrimidine derivatives 1 and 2 can be synthesized according to literature procedures (J. Org. Chem., 26, 2755, 1961).

DETAILED DESCRIPTION OF THE INVENTION

Example A

Preparation

CO 2 H • 2HCl

5-Aminonicotinic acid (4.0 g, 0.021 mol) (Helv. Chim. Acta, 47, 363 (1964); JACS, 70, 2381 [1948] with 1H-pyrazole-1-carboxamidine hydrochloride (4.6 g) , 0.031 mol), diisopropylethylamine (8.0 g, 0.062 mol) and dioxane (14 ml) and H ₂ O (7 ml) were heated at reflux for 2 days, the reaction was cooled to room temperature, the precipitate was filtered, washed with H ₂ O / dioxane (50:50) and dried The precipitate was suspended in H ₂ O and acidified with 2M HCl The solvent was removed in vacuo to give the title compound as a white solid (750 mg).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Example B

Preparation

co ₂ h

2HCI ······ · · · ····· · · · · · ·

Step 1

To 3,4,5,6-terahydro-2-pyrimidinethiol (10 g, 0.086 mol) (Aldrich) in absolute ethanol (75 mL) was added Mel (12.2 g, 0.086 mol). The reaction was stirred at reflux for 2.5 hours. The solvent was removed in vacuo and the product dried to give 2-methylthio-3,4,5,6-terahydro-2-pyrimidine hydroiodide as a white solid (22 g).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 2

To the product of Step 1 above (5.3 g, 0.021 mol) and triethylamine (2.07 g, 0.021 mol) in CH ₂ Cl ₂ (25 mL) was added BOC anhydride (4.5 g, 0.021 mol) at an ice bath temperature. The reaction was then stirred at room temperature for 2 days. CH ₂ Cl ₂ was washed with H ₂ O (3x), dried over MgSO 4, and removed in vacuo to give N-Boc-2-methylthio-3,4,5,6-tetrahydro-2-pyrimidine (4.14 g) .

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 3

The product from Step 2 above (3.08 g, 0.0134 mol) was heated at 80 ° C to 85 ° C with 5-aminonicotinic acid (1.8 g, 0.0134 mol) in DMA (12 mL) 2 weeks. The reaction mixture was diluted with CH ₃ CN, the precipitate was filtered, washed with CH ₃ CN and dried. The precipitate was suspended in H ₂ O and the pH was adjusted to 1-2 with concentrated HCl. The solution was frozen and lyophilized to give the desired product as a light brown solid (1.2 g).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Example C

Preparation

Step 1

To 2-methylthio-2-imidazoline hydrochloride (20 g, 0.082 mol) (Aldrich) and triethylamine (8.28 g, 0.082 mol) in CH ₂ Cl ₂ (100 mL) was added BOC anhydride (17.9 g, 0.082 mol) mol) ······ 4 4 4 4 · 4 • β 4 4 4 · · · · ·

444 444 · 44 4 ··· 444444 4 4 • 444 44 4 44 4 • 4 44 44 · ·· · · · at an ice bath temperature. The reaction was then stirred overnight at room temperature. The CH ₂ Cl ₂ was washed with H ₂ O (2x), dried over MgSO ₄ and removed in vacuo to give N-BOC-2-methylthio-2-imidazoline as a viscous oil which gave a waxy white solid (15, 93 g).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 2

The product of Step 1 above (15.93 g, 0.0737 mol) was treated with 5-aminonicotinic acid (9.6 g, 0.07 mol) and triethylamine (7.1 g, 0.07 mol). in DMA (60 mL) was heated at 100 ° C for 2 days, then at 130 ° C for 1 day and at 150 ° C for 2 days. After cooling and diluting the reaction mixture with CH ₃ CN, the precipitate was filtered, washed with ether and dried (zwitterion yield 10.16 g). This product was suspended in H ₂ O and acidified with TFA (pH 1-2). The solution was frozen and lyophilized to give the desired product as a pale yellow solid (20.15 g).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Example D

Preparation

Η ₂ Ν · ^ ηΓ ^Ν

O

COzEt

OH

HCl

Cl

Cl ···· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

Step 1

Preparation • · · · · · · · · · · · · · · ·

Whose

To a 2 L round bottom flask equipped with a mechanical stirrer and condenser was added 3,5-dichlorosalicylaldehyde (200.0 g, 1.05 mol, 1 equivalent), acetic anhydride (356 g, 3.49 mol) and triethylamine (95 0.9 g, 0.94 mol, 0.90 equivalents). The reaction solution was heated at reflux overnight. The dark brown reaction mixture was cooled to 50 ° C and water (1 L) was added with stirring. After an hour, the mixture was filtered and the filtrate was mixed with EtOH (11). The mixture was heated at 45 ° C for one hour, cooled to room temperature, filtered and the solid (fraction A) was washed in EtOH (0.5 L). The combined EtOH solutions were concentrated on a rotary evaporator to an oil (fraction B). The solid from fraction A was dissolved in dichloromethane (1.5 L) and the resulting solution was purified over silica gel (1300 mL volume). The resulting dark brown solution was concentrated to an oil which was mixed with hexane (1.3 L) to give a solid which, after isolation by filtration and washing (hexane), yielded substantially pure 6,8-dichlorocoumarin (163 g). An additional 31 g of product was obtained by treating the oil (fraction B) in the same manner; the oil was dissolved in dichloromethane (0.5 L), purified through silica gel (0.5 L volume), and mixed with hexane. The total amount isolated was 194 g or 86% of a brown solid.

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 2 Preparation

To a 3 neck 2 L round bottom flask equipped with a mechanical stirrer and condenser was added 6,8-dichlorocoumarin (160 g, 0.74 mol) prepared in Step 1 and dry THF (375 mL, Aldrich Sure Seal). The resulting mixture was cooled to -40 ° C (dry ice in acetone bath) and lithium bis (trimethylsilyl) amide (0.80 mol, 800 mL, 1M in THF) was added while maintaining the temperature below -40 ° C. After all components were added, the cooling bath was removed. After 0.5 h, the mixture was warmed to -5 ° C. The reaction was stopped by adding a solution of HCl (0.5 14M dioxane) in EtOH (1.25 L). The temperature was kept below 0 ° C overnight. The reaction mixture was concentrated to approximately half the original volume and partitioned between EtOAc (3 L) and water (2 L). The organic layer was washed with water HCl (3 x 11, 0.5 M HCl). The pH of the combined aqueous layers was adjusted to about 7 by addition of 10% aqueous NaOH and extraction was performed in dichloromethane (3 x 2 L). The combined organic layers were dried (MgSO ₄ ), filtered, and HCl (210 mL, 4M in dioxane) was added with stirring. After precipitation, the solid was collected by filtration. The filtrate was concentrated to a small amount and methyl t-butyl ether was added. The obtained solid was mixed with the initially formed solid and the combined product was washed in methyl t-butyl ether, isolated by filtration and dried (in a vacuum oven for two days) to give the desired product (172 g, 74% yield).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 3 Preparation

Nt-Boc-glycine N-hydroxysuccinimide ester (Sigma, 15.0 g, 0.055 mol), dry DMF (Aldrich Sure Seal, 200 mL) was added to a flame-dried round bottom (0.5 L) flask equipped with a magnetic stirrer. and the product of Step 2 (21.67 g, 0.055 mol) under an inert atmosphere (Ar). The reaction mixture was cooled to approximately 0 ° C (ice bath with salt) and N-methylmorpholine (5.58 g, 0.056 mol) and a catalytic amount of DMAP were added. The reaction was allowed to proceed overnight. The reaction mixture was concentrated to a slurry and partitioned between EtOAc (0.4 L) and aqueous base (2 χ 2 L, aqueous saturated NaHCO ₃ ). The organic layer was washed successively with aqueous citric acid (2 χ 0.2 L, 10% w / v), aqueous sodium bicarbonate (2 χ 0.2 L), brine and dried (Na ₂ SO ₄ ). The volatiles were removed under vacuum at 55 ° C to give an oil (22.5 g, 92% yield) which solidified on standing.

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 4 Preparation

I i

This it this it this it this it this it it it it it to it to ···

The product of step 3 was deprotected and the hydrochloric acid salt was obtained according to the following procedure. To the product obtained in Step 3 (14.0 g, 0.032 mol) was added dry dioxane (40 mL) in a flame dried round bottom flask (0.1 L) with stirring with a glass rod. To this solution was added HCl (4.0 M in dioxane, 2 equivalents, 6.32 mL) at 0 ° C. The reaction was complete until gas evolution had ceased completely. The volatiles were removed in vacuo and the residue was triturated in diethyl ether (50 mL). The solids were combined by filtration, washed with ether and dried to give the desired product (12.5 g).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Example E

Preparation

Step 1 Preparation

To a suspension of 3-bromo-5-chlorosalicylaldehyde (175 g, 743.2 mmol) in acetic anhydride (280.5 mL, 3 mol) was added triethylamine (103.6 mL, 743.2 mmol). The reaction solution was heated at reflux for 4.5 hours. The solution was cooled and concentrated in vacuo. The brown residue was added to absolute ethanol (730 mL). The mixture was stored at 0 ° C for 14 hours.

······ · · · φ · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · Φφφ • Φ φφ φφ φ φφ φφφ

Filtration gave a brown solid which was washed with chilled ethanol. Drying the solid under vacuum gave the desired product (123 g, 64% yield).

The H ¹ -NMR measurement was consistent with the desired structure.

Step 2 Preparation

To a suspension of the coumarin obtained in Step 1 (40.0 g, 154.1 mmol) in THF (400 mL) at -76 ° C was added dropwise lithium bis (trimethylsilyl) amide (154.1 mL of a 1M solution in THF). The addition took 10 minutes, then the reaction mixture was stirred for 5 minutes, warmed to -20 ° C and stirred for 15 minutes. To this solution was added acetic acid (9.25 g, 154.1 mmol) in THF (28 mL) over 5 minutes. The mixture was warmed to room temperature and the volatiles were removed in vacuo. The residue was dissolved in ether (850 mL), washed with saturated aqueous NaHCO ₃ (2 × 100 mL), brine (2 × 40 mL), and dried (MgSO 4). The ether solution was concentrated to about 160 mL and cooled to 0 ° C. To this suspension was added HCl (4M in dioxane, 56.3 mL, 225 mmol) and the mixture was stirred at 0 ° C for 30 min. The suspension was filtered and the filter cake was washed thoroughly with ether. The solid was dried in vacuo to give the desired product as the HCl salt, dioxane solvate (45 g). The H ¹ -NMR measurement was consistent with the desired structure.

· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

Step 3 Preparation

To a suspension of the lactone produced in Step 2 (142.2 g, 354.5 mmol) in absolute ethanol (533 mL) was added HCl (4M in dioxane, 157.8 mL, 631.1 mmol) over 10 minutes. The reaction mixture was stirred at room temperature for 2.5 hours. The volatiles were removed under vacuum. The residue was dissolved in ethyl acetate (450 mL) and the solution was stored at 0 ° C for 15 h. A tan precipitate was obtained by filtration and washed with chilled ethyl acetate. The solid was dried under vacuum to give the desired product as the hydrochloride salt (100.4 g, 79% yield).

The H ¹ -NMR measurement was consistent with the desired structure.

Step 4 Preparation

Nt-Boc-glycine N-hydroxysuccinimide ester (Sigma, 2.72 g, 0.010 mol), dry THF (Aldrich, Sure, Seal, 50 mL) was added to a flame dried round bottom flask (0.1 L) equipped with a stirrer. and the product of Step 3 (3.10 g, 0.01 mol, vacuum dried over P2O5 overnight) under an inert atmosphere (Ar). The reaction mixture was cooled to approximately 0 ° C (ice bath with salt) and triethylamine (1.01 g, 0.010 mol) was added. The reaction was carried out overnight. The reaction mixture was concentrated to a semi-solid and worked up in a manner similar to that of Example A, Step 3. The volatiles were removed from the organic

0 · 0 · «0 00 · 00 ·

000 000 «0 000 000 0000 00 0

0 0 0 0 0 0000 0 0 0 0

0000 00 00 00 00

The layer was vacuum dried at 55 ° C to give an oil (4.0 g, 83% yield) which solidified on standing.

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 5 Preparation

The product obtained in Step 4 was deprotected and the hydrochloride salt was obtained using the following procedure. To the product obtained in Step 4 (4.0 g, 0.0084 mol) was added dry dioxane (20 mL) in a flame dried round bottom flask (0.1 L) while stirring with a glass rod. To this mixture was added HCl (4M in dioxane, 20 mL) and the reaction continued until gas evolution was complete (approximately 1 hour). The volatiles were removed in vacuo and the residue was mixed with diethyl ether (50 mL). The solids were collected by filtration, washed with ether, and dried to give a light brown solid (2.7 g, 78% yield).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Example F

Preparation

Cl

CQřEt

OH • HCI 99 9 9

Step 1

Preparation of 3-iodo-5-chlorosalicylaldehyde

Cl

H

O

OH

To a solution of 5-chlorosalicylaldehyde (100 g, 0.638 mol) in dimethylformamide (400 mL) was added N-iodosuccinimide (144.0 g, 0.641 mol). The reaction mixture was stirred at room temperature for 2 days. Additional N-iodosuccinimide (20.0 g) was added and stirring was continued for another two days. The reaction mixture was diluted with ethyl acetate (11), washed with hydrochloric acid (300 mL, 0.1 M), water (300 mL), sodium thiosulfate (5%, 300 mL), brine (300 mL), dried (MgSO ₄ ) and concentrated to dryness to give the desired aldehyde as a pale yellow solid (162 g, 90% yield).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 2

Preparation of 6-chloro-8-iodocoumarin

A mixture of 3-iodo-5-chlorosalicylaldehyde (100 g, 0.354 mol), acetic anhydride (300 mL) and triethylamine (54 mL) was heated at reflux for 18 hours. Upon cooling, the desired coumarin precipitated as a dark brown crystalline solid. The precipitate was filtered, washed with hexane / ethyl acetate (4: 1, 200 mL) and air dried. [Yield: 60 g (55% yield)]. Additional amounts of the desired product (10 g, 9% yield) can be obtained from the filtrate during storage.

MS and H ¹ -NMR measurements were consistent with the desired structure.

9

Preparation of (R, S) -4-amino-3,4-dihydro-6-chloro-8-iodocoumarin hydrochloride • β · 9 9 · 9 9 9 9 ··· · · · · 9 9 9

999 9999999 9 9

9 9 9 9 9

9 9 9 9 9 9 9 9 9

Step 3

To a solution of 6-chloro-8-iodocoumarin (6.63 g, 21.62 mmol) in tetrahydrofuran (100 mL) at -78 ° C was added lithium hexamethyldisilazane (21.62 mL, 1M, 21.62 mmol). The reaction mixture was stirred at this temperature for 30 minutes and then at 0 ° C for 1 hour. Acetic acid (1.3 g, 21.62 mmol) was added to the reaction mixture. The reaction mixture was poured into ethyl acetate (300 mL) and saturated with sodium carbonate solution (200 mL). The organic layer was separated, and washed with brine (200 mL) followed by dioxane / HCl (4M, 30 mL) at 0 ° C. The reaction mixture was stirred for 1 hour at room temperature, filtered and dried under vacuum to afford the desired product as a powder (4.6 g, 59% yield). (RPHPLC: _Rf 6.8 minutes gradient of 10% acetonitrile - 90% acetonitrile in 15 min, then 100% acetonitrile in 6 min. Both water and acetonitrile contained 0.1% TFA Protein peptide column Vydac C18, flow rate 2 ml / min, monitored at 254 nm).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 4

Preparation of (R, S) -ethyl- (3-amino-3- (5-chloro-2-hydroxy-3-iodo) phenylpropionate) hydrochloride.

NH ₂

COzEt

OH

Cl «· · · ··· ·

A solution of 4-amino-3,4-dihydro-6-chloro-8-iodocoumarin hydrochloride (22.0 g, 61.09 mmol) in ethanol (250 mL) was bubbled through hydrogen chloride at 0-10 ° C until saturation. . After 6 hours at reflux, most of the solvent was removed by distillation. The cooled residue was added to anhydrous ether and stirred for 2 hours. The original gummy consistency was converted to a crystalline substance. The crystalline product was filtered and dried to give the desired product as an off-white crystalline powder (20 g, 81% yield). (Rf, 7.52 min, conditions as in step 3).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 5

Preparation of (R, S) -ethyl- (3- (N-BOC-gly) -amine-3- (5-chloro-2-hydroxy-3-iodo) phenylpropionate)

A mixture of BOC-gly (2.16 g, 12.31 mmol), HOBT (1.67 g, 12.31), EDCI (2.36 g, 12.31 mmol) and DMF (50 mL) was stirred for 1 hour at 0 ° C. To the reaction mixture was added ethyl (3-amino-3- (5-chloro-2-hydroxy-3-iodo) -propionate) hydrochloride (5.0 g, 12.31 mmol) followed by triethylamine (3.5 mL). The reaction mixture was stirred at room temperature for 18 hours. The DMF was removed in vacuo and the residue was partitioned between ethyl acetate (300 mL) and sodium bicarbonate (200 mL). The organic layer was washed with hydrochloric acid (1M, 100 mL), brine (200 mL), dried (MgSO ₄ ) and concentrated to give the desired product as a solid (6 g, 93% yield).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Step 6

Preparation of (R, S) -ethyl- (3- (N-gly) -amino-3- (5-chloro-2-hydroxy-3-iodo) phenylpropionate) hydrochloride

• · · · · · • 9 · • · fe · · 9 9 · 9 9 9 9 9 • · · 99 9999999 9 9 • · «9 9 99 9

To ethyl 3- (N-BOC-gly) -amine-3- (5-chloro-2-hydroxy-3-iodo) propionate (6.0 g, 11.30 mmol) was added dioxane / ethyl acetate at 0 ° C. HCl (4M, 2 mL) and stirred at room temperature for 3 hours. The reaction mixture was concentrated, toluene (100 mL) was added, and then the reaction mixture was concentrated again. The obtained residue was suspended in ether, filtered and dried to give the desired product as a crystalline powder (5.0 g, 95% yield), (rphplc: Rf 8.3 min, conditions as in step 3).

MS and H ¹ -NMR measurements were consistent with the desired structure.

Example G

Preparation

Step 1

Preparation of Reformatski

BrZnCH ₂ CO2-t-Bu

A 4 liter flask equipped with a condenser, thermometer, and mechanical stirrer was charged with Zn metal (180.0 g, 2.76 mol, 30-100 grain size) and THF (1.25 L). While stirring, 1,2-dibromoethane (4.74 ml, 0.06 mol) was added via syringe [alternatively, it can be replaced by the addition of TMS Cl (0.1 equivalents) over one hour at ····· · · · 9 9 »

9 9 9 9 9

9 9 9 9 9

9,999 9999999 9 9

9 9 9 9 9

99 99 * 99 99 9 room temperature]. After purging with inert gas (3 cycles of N ₂ / vacuum), the zinc suspension in THF was heated to reflux (65 ° C) and left at this temperature for 1 hour. The mixture was cooled to 50 ° C before addition of tert-butyl bromoacetate (488 g, 369 mL, 2.5 mol) using a 50 mL syringe and syringe pump (addition rate 4.1 mL / min) over 1.5 hours. The reaction temperature was maintained at 50 ° C +/- 5 ° C during the addition. After the addition was complete, the reaction mixture was subsequently stirred at 50 ° C for 1 hour. Then the reaction mixture was cooled to 25 ° C and the precipitated product settled. The mother liquor was decanted into a two-neck round bottom flask using coarse filter paper and partial vacuum (20 mm Hg). This removed approximately 65% THF from the mixture. 1-Methyl-2-pyrrolidinone (NMP, 800 mL) was added and the reaction continued for 5 minutes. The reaction mixture can be filtered to remove residual zinc. The titer of the desired Reformatski reagent was determined to be 1.57 M with a molar yield of 94%. Optionally, the solid may be isolated by filtration from the original reaction mixture. The filter cake was washed with THF until a white solid was obtained and dried under a nitrogen atmosphere. The desired product was obtained as a mono solvate of THF, which can be stored at -20 ° C (recommended) for extended periods. Typical yields are 85-90%.

Step 2 A. Preparation

To a solution of 3,5-dichlorosalicylaldehyde (11.46 g, 60 mol) in DMF (40 mL) was added potassium carbonate (powder dried in an oven at 100 ° C under vacuum, 8.82 g, 60 mmol) at room temperature. to give a pale yellow suspension. MEMCI (pure, 7.64 g, 61 mmol) was then added while maintaining the bath temperature at 20 ° C. The mixture was then stirred at 22 ° C for 6 hours and MEMCI (0.3 g, 2.4 mmol) was added. The mixture was stirred for an additional 0.5 h and the reaction mixture was poured into cold water (200 mL) to precipitate the product. The suspension was filtered on a pressure filter and the filter cake was washed with water (2 x 50 mL) and dried under N ₂ / vacuum to give the product as an off-white solid. (14.94 g, 89% yield).

¹ H NMR (CDCb, TMS) 3.37 (s, 3H). 3.54 to 3.56 (m, 2H), 3.91 to 3.93 (m, 2H), 5.30 (s, 2H), 7.63 (d, 1H), 7.73 (d, 1H), 10.30 (s, 1H); ¹³ C NMR (CDCl ₃ , TMS) δ (ppm): 59.03, 70.11, 99.57, 126.60, 129.57, 130.81, 132.07, 135.36, 154.66, 188.30. DSC: 48.24 ° C (endo 90.51 J / g).

Microanalysis: calculated for C 11 H 12 Cl 2 O 4: C: 47.33%; H, 4.33%; Cl: 25.40% True: C: 47.15%; H. 4.26%; Cl: 25.16%.

The product of Step 2A (35.0 g, 0.125 mol) was added to a 1 L 3-neck round bottom flask equipped with a mechanical stirrer and additional funnel to add THF (200 mL). The solution was stirred at 22 ° C and (S) -phenylglycinol (17.20 g, 0.125 mol) was added in one portion. After 30 minutes, MgSO ₄ (20 g) was added at 22 ° C. The mixture was stirred at 22 ° C for 1 hour and filtered through coarse filter paper. The filtrate was concentrated under reduced pressure. No further purification was performed and the crude imine was used directly in the condensation reaction, in Step 2 of Example C.

· · T · t t * t t t t t t t t t t t t φ φ φ φ

The product of Step 1 (91.3 g, 0.275 mol) and NMP (200 mL) under nitrogen was added via addition funnel to a three necked 1 liter round bottom flask equipped with a mechanical stirrer. The solution was then cooled to -10 ° C and stirred at 350 rpm. A solution of the imine (prepared in Step B) in NMP was prepared under nitrogen and then added to the reaction mixture over 20 minutes while maintaining the temperature at -5 ° C (jacket temperature -10 ° C). The mixture was stirred for an additional 1.5 hours at -8 ° C and after the addition was completed for an additional hour at -5 ° C. After cooling to -10 ° C, concentrated HCl / saturated NH ₄ Cl solution (100 mL), water (100 mL) was added, and brine (100 mL) was added to the solution. MTBE (200 mL) was added and the mixture was stirred for 15 minutes at 23 ° C and 200 rpm. Stirring was stopped and the layers were separated. The aqueous layer was extracted with MTBE (100 mL). The two organic layers were combined, washed sequentially with saturated NH ₄ Cl solution (100 mL), water (100 mL), and brine (100 mL). The solution was dried with MgSO ₄ (30 g), filtered and concentrated to give an orange oil (66.3 g) (solidified on standing) containing the desired product as a simple diastereoisomer as determined by NMR protons and carbons. The sample was purified by recrystallization from heptane for analysis. This gave the product as an off-white solid. Proton and carbon NMR and IR spectra were consistent with the desired product. [α] ^D 25 = + 8.7 ° (c = 1.057, MeOH). Microanalysis: calculated for C ₂₅ H ₃ 3Cl ₂ NO ₆ :

C: 58.77%; H: 6.47%; N: 2.72%; Cl: 13.78%

Found: C: 58.22%; H: 6.54%; N: 2.70%; Cl: 13.66% * 9 · »·»

Step 3 Preparation

A solution of the crude ester prepared in Step 2 [17.40 g, 0.033 mol (in theory)] and EtOH (250 mL) was added to a 1 liter three necked flask reactor. The solution was cooled to 0 ° C and Pb (OAc) ₄ (14.63 g, 0.033 mol) was added in one portion. After 2 hours, a 15% NaOH solution (30 mL) was added and the ethanol was removed under reduced pressure. Another portion of 15% NaOH (100 mL) was added and the mixture was extracted with MTBE (2 100 mL), washed with H ₂ O (2 χ 100 mL) and brine (50 mL), dried over Na ₂ SO ₄ , filtered through celite and It was concentrated under reduced pressure to give an orange oil (12.46 g) as a homogeneous solid after determination of the body. The oil was used without further purification.

B. The oil from Step A was diluted with EtOH (30 mL) and paratoluenesulfonic acid (1.3 equivalents, 0.043 mol, 8.18 g) was added. The solution was heated to reflux for 8 hours, cooled to ambient temperature and concentrated under reduced pressure. The residue was mixed with THF (20 mL) and heated to reflux to form a solution. The solution was cooled to room temperature and the compound crystallized. Heptane (30 mL) and THF (10 mL) were added and a liquid suspension formed which was filtered. The filter cake was washed with THF / heptane (40 mL, 1/1), dried under vacuum on a pressure filter under nitrogen atmosphere for two hours to give a white solid (7.40 g).

Proton and carbon NMR and IR spectra were consistent with the desired product essentially in the form of one enantiomer.

Microanalysis: Calculated for C ₈ H ₂₁ Cl ₂ NO ₆ S, 0.25 C ₄ H ₈ O:

C: 48.73%; H, 4.95%; N: 2.99%; Cl: 15.14%

Found: C: 48.91%; H, 4.95%; N: 2.90%; Cl: 14.95% • e 0 · 0 t ·

000 000 000

000 0000 0 0

000 0000000 ·

0 »0 · 0 0 · 00 00 ·

Step 4 Preparation

A round bottom flask (500 mL) equipped with a magnetic stirrer was charged under N 3 with 21.7 g (0.065 mol), Nt-Bocglycine N-hydroxysuccinimide ester (17.7 g, 0.065 mol) and DMF ( 200 ml). The reaction mixture was stirred under a nitrogen atmosphere at room temperature for 3.25 hours to give a pale orange solution. The reaction mixture was poured into ice-cooled ethyl acetate (1.2 L). The organic solution was washed with 1M HCl (250 mL) and then brine (500 mL), dried (MgSO ₄ ) and concentrated to near dryness in vacuo to give an oil which was then dried at 50 ° C to give the product as a colorless oil. (28.12 g, 99% yield). Granular crystals were prepared from ethyl acetate / hexane. The product (approximately 28 g) was dissolved in ethyl acetate (35 mL) and hexane (125 mL). The granular crystal solution crystallized to form a precipitate. The solids were filtered and dried under vacuum at 55 ° C overnight to give a colorless solid (27.0 g, 95% yield). ¹ H-NMR was consistent with the desired product.

Step 5 Preparation

CO ₂ Et

Whose

Η ₂ Ν ^ γ ^Ν

OH * HCl

Cl totototo · this to · to · to · this ·· ··········

To to to · ·....

The Boc protected glycamide prepared in Step 4 (27.0 g, 0.062 mol) was dried over P ₂ O 5 overnight and the NaOH settled. The solid was dissolved in dioxane (40 mL) and the solution was cooled to 0 ° C. An equivalent volume of 4M HCl / dioxane (0.062 mol) was added and the reaction was allowed to proceed for 2 hours. During this time, the conversion was 80% as determined by rphplc. The reaction mixture was warmed to room temperature for 4 hours. The reaction mixture was concentrated to a foam at 40 ° C which was stirred in ether (200 mL). A white solid formed which was filtered and dried over P ₂ O ₅ . The desired glycine beta-amino acid ethyl ester was obtained as the HCl salt (20.4%, 88.5% yield after isolation).

¹ H-NMR and MS were consistent with structure.

Example H

Preparation

Step 1 Preparation

To a solution of 3-chloro-5-bromosalicylaldehyde (35 g, 0.15 mol) in DMF (175 mL) was added potassium carbonate (powdered, oven dried at 100 ° C under vacuum, 22.1 g, 0.16 mol). ) at room temperature to give a pale yellow suspension. MEMCI (neat, 25.0 g, 0.2 mol) was then added and the bath temperature was maintained at 20 ° C. The mixture was then stirred for 6 hours. 4 4 4 4 4 4 9

9 9 9 4 9 4

4 4 4 4 4444494 »

ZTQ 4 4 · 4 4 9 4 9 4

VS 44 49 44 at 22 ° C and poured into DI water (1200 mL) and the product precipitated. The suspension was filtered on a pressure filter, the filter cake was washed with DI water (2 2 400 mL) and dried under vacuum in a nitrogen atmosphere to give the product as an off-white solid (46 g, 95% yield). ¹ H-NMR (CDCl 3, TMS) 3.35 (s, 3H), 3.54-3.56 (m, 2H), 3.91-3.93 (m, 2H), 5.30 (s, 2H), 7.77 (d, 1H), 7.85 (d, 1H), 10.30 (s, 1H); ¹³ C NMR (CDCl3, TMS)? (Ppm): 59.05, 70.11, 71.49, 99.50, 117.93, 129.69, 129.78, 132.37, 138.14, 155, 12, 188.22. DSC: 48.24 ° C (endo 90.51 J / g);

Microanalysis: calculated for C 11 H 15 BrClO 4

C: 40.82%; H, 3.74%; N: 10.95%; Br: 24.69% True: C: 40.64%; H, 3.48%; N: 10.99%; Br: 24.67%

Step 2 Preparation

The product obtained in Step 1 (32.35 g, 0.1 mol) was added to a 500 mL three-necked round bottom flask equipped with a mechanical stirrer followed by THF (160 mL) and (S) -phenylglycinol (13.71 g). , 0.1 mol). After 30 minutes, MgSO ₄ (20 g) was added at 22 ° C. The mixture was stirred at 22 ° C for 1 hour and filtered through coarse filter paper. The filtrate was concentrated under reduced pressure to give a pale yellow oil (48.0 g) containing imine. The crude product without further purification was used directly in the coupling reaction. Microanalysis: Calcd for Ci9H ₂ iBrCINO _4:

C: 51.54%; H, 4.78%; N: 3.16%; Br: 18.04%; Cl: 8.00%;

C: 51.52%; H: 5.02%; N: 2.82%; Br; 16.31%; Cl: 7.61%.

Actual values:

Step 3

Preparation 99 ··· 99 99

9 9 9 • · · 9

9 9 9 9 • 9 9 9 9 9

9 · 99 99 9

To a 3L necked round bottom flask equipped with a mechanical stirrer was added the reagent from Step 1 of Example G (332 g, 0.8 mol) in NMP (660 mL) under a nitrogen atmosphere. The solution was then cooled to -10 ° C. Under a nitrogen atmosphere, a solution of the imine produced in step 2 in NMP (320 mL) under nitrogen was prepared and then added to the above reaction mixture over 30 minutes while maintaining the temperature at -5 ° C. The mixture was stirred for an additional hour and then cooled to -10 ° C. A mixture of concentrated NH ₄ Cl solution (30 mL / 720 mL) saturated with HCl was added over 10 minutes. MTBE (760 mL) was added and the mixture was stirred at 23 ° C for 1 h. Stirring was stopped and the layers were separated. The aqueous layer was extracted with MTBE (320 mL). The organic layers were combined, washed thoroughly with saturated NH ₄ Cl solution (320 mL), DI water (320 mL), and brine (320 mL). The solution was dried with MgSO ₄ (60 g), and filtered and concentrated to give a yellow oil (228 g) containing the desired product as one diastereoisomer.

DSC: 227.54 ° C (endo, 61.63 J / g);

Microanalysis: calculated for C ₂₅ H33BrCINO6:

C: 53.72%; H: 5.95%; N: 2.50%; Br: 14.29%; Cl: 6.33%

Found: C: 53.80%; H: 6.45%; N: 2.23%; Br: 12.85%; Cl: 6.12%.

4445 44 • 4

Step 4

Preparation • 4 4 4 • 4 4 4

44 ·· • • 4 <

4 4 4 • 4 4444 • · 4 · 4

A solution of the crude ester from step 3 (about 111 g) in ethanol (1500 mL) was added under nitrogen to a 3L three-necked round bottom flask equipped with a mechanical stirrer. The reaction mixture was cooled to 0 ° C and lead tetraacetate (88.67 g, 0.2 mol) was added in one portion. The reaction mixture was stirred at 0 ° C for 3 hours and then 15% NaOH (150 mL) was added to the reaction mixture at a temperature below 5 ° C. Ethanol was removed on a rotary evaporator. Additional 15% NaOH (600 mL) was added and the reaction mixture was extracted with ethyl acetate (2 x 300 mL), MTBE (2 x 200 mL) and ethyl acetate (2 x 200 mL). The organic layers were combined and washed with DI water (2 x 200 mL) and brine (2 x 100 mL) and dried over MgSO ₄ (30 g). The solution was then filtered through celite and concentrated under reduced pressure to give the product as an orange oil (96 g), which was used in the next step without further purification.

DSC: 233.60 ° C (endo. 67.85 J / g);

Microanalysis: calculated for C ₂₄ H ₂ gBrCINO ₅ :

C: 54.71%; H: 5.54%; N: 2.65%; Br: 15.16%; Cl: 6.72%;

Found: C: 52.12%; H: 5.40%; N: 2.47%; Br: 14.77%; Cl: 6.48%.

Step 5

Preparation

To the crude product from Step 4 (about 94 g) in absolute ethanol (180 mL) was added paratoluenesulfonic acid monohydrate (50.0 g, 0.26 mol). The reaction mixture was heated to reflux for 8 hours and the solvent was then removed under reduced pressure. The solid residue was mixed with THF (100 mL) and then THF was removed under reduced pressure. The residue was dissolved in ethyl acetate (500 mL) and cooled to about 5 ° C. The resulting solid was filtered and washed in heptane (2 × 50 mL) to give a white solid. The solid was air dried to give the product as a white solid (38 g) as a single isomer.

¹ H - NMR (DMSO, TMS)? (Ppm) 1.12 (t, 3H), 2.29 (s, 3H), 3.0 (m, 2H), 4.05 (q, 2H), 4, 88 (t, 1H), 7.11 (d, 2H), 7.48 (d, 2H), 7.55 (d, 1H), 7.68 (1H, d); 8.35 (br. S, 3H); ¹³ C NMR (DMSO, TMS) (ppm): 13.82, 20.75, 37.13, 45.59, 60.59, 110.53, 122.47, 125.44, 127.87, 128, 06, 129.51, 131.95, 137.77, 145.33, 150.14, 168.98; DSC: 69.86 ° C (end., 406.5 J / g), 165.72 ° C (end. 62.27 J / g), 211.24 ° C (exo. 20.56 J / g) [α] D ₂₅ = + 4.2 ° (c = 0.960, MeOH); IR (MIR) (cm ^-1 ) 2922, 1726, 1621, 1591, 1494, 1471, 1413, 1376, 1324, 1286, 1237, 1207;

Microanalysis: calculated for C ₁₈ H ₂ iBrCINO6S:

C: 43.69%; H: 4.27%; N: 2.83%; Br: 16.15%; Cl: 7.16%; S: 6.48%;

Found: C: 43.40%; H: 4.24%; N: 2.73%; Br: 16.40%; Cl: 7.20%; S: 6.54%.

• · • ·

Step 6

Preparation ·· · ·

• KCl

The above compound was prepared according to the procedures of Example G, Step 4 and Step 5, in which an equivalent amount of the intermediate prepared in Step 5, as the free base, was replaced with the intermediate from Step 4 of Example G.

1 H-NMR and MS were consistent with the desired product.

Example I

Preparation

• HCI • 999 • 99 • 99 • 9 • 99 999 • 99 9999 9 9

999 9 9999 99 9 •• 99 99 9 99 • 99 99 9 99 9

Step 1

Preparation

The above compound was prepared according to the procedure for Example G, step 2A, substituting an equivalent amount of 2-hydroxy-3,5-dibromobenzaldehyde with 3,5-dichlorosalicylaldehyde.

Yield: 88%; pale yellow solid: mp 46-47 ° C; R _f = 0.6 (EtOAc / hexane 1: 1 v / v); ¹ H-NMR (CDCl ₃ ) δ 3.37 (s, 3H), 3.56 (m, 2H), 3.92 (m, 2H), 5.29 (s, 2H), 7.91 (d 1 H, J = 2.4 Hz), 7.94 (d, 1H, J = 2.4 Hz), 10.27 (s, 1H); FAB-MS m / z 367 (MH +).

(M ⁺ ) HR - MS calcd for C 11 H 11 B 2 O 4 367.9083 Found: 367.9077.

¹ H-NMR and MS were consistent with the desired compound.

Step 2

The above compound was prepared according to the procedure in Example G, steps 2B and 2C, by replacing an equivalent amount of the compound in step 1 of the process.

Yield: 90%; yellow solid; mp 57-59 ° C; _Rf = 0.46 (EtOAc / hexane 1: 1 v / v); ¹ H-NMR (CDCl ₃ ) δ 1.45, (s, 9H); 2.1 (br, 1H, exchangeable), 2.51 (d, 1H, J ₃ = 9.9 Hz, J ₂ = 15.3 Hz), 2.66 (d, 1H, J ₃ = 4.2 Hz, J ₂ = 15.3 Hz), 3.02 (br, 1H, exchangeable), 3.39 (s, 3H), 3.58 - 3.62 (m, 4H), 3.81 (m, 1H), 3.93 (m, 2H), 4.63 (dd, 1H, J = 4.2Hz), 5.15 (s, 2H), 7.17-7.25 (m, 6H), 7.49 (d, IH): FAB-MS m / z 602 (M + H).

HR-MS calc for C ₂₅ H ₃₄ N ₂ O ₂ O ₆ : 602.0753

Found: 602.0749 ¹ H-NMR and MS were consistent with the desired compound.

Step 3 Preparation

The above compound (p-toluenesulfonate salt) was prepared according to the procedure of Example G, Step 3, by substituting an equivalent amount of the product in Step 2 of the Example.

G, step 3A. Yield: 62%; white solid; ¹ H-NMR (DMSO-d ₆ ) δ 1.09, (t, 3H, J =

7.2 Hz); 2.27 (s, 3H), 2.97 (dd, 2H, J = 3.0 Hz, J ₂ = 7.2 Hz); 4.02 (q, 2H, J = 7.2 Hz), 4.87 (t, 1H, J = 7.2 Hz), 7.08 (d, 2H, J = 4.8 Hz), 7 45 (m, 3H), 7.57 (d, 1H, J = 2.4Hz),

8.2 (br. 3H); FAB-MS m / z 365 (M + H) &lt; + &gt;

HR-MS calc for C ₁₁ H ₁₄ N ₂ O ₂ O 3: 365.9340

Found: 365.9311 ¹ H-NMR and MS were consistent with the desired product.

Step 4 Preparation

• HCl

The above compound was prepared according to the procedure in Example G, Step 4, and replacing the compound prepared in Step 3 to prepare the BOC protected intermediate. The resulting BOC protected intermediate was converted to the desired compound using the procedure of Example G, Step 5.

¹ H-NMR and MS were consistent with the desired compound.

• *

Example J

Preparation of the bis hydrochloride salt of ethyl 3 - [(2-aminoacetyl) amino] pyridine-3-propanoate

Η ₂ Ν ^ γ

O

CO ₂ Et

2HCI

Step 1

To 3-pyridine carboxaldehyde (300 mL) in 2-propanol (3 L) was added ammonium acetate (297 g) followed by malonic acid (398 g). The reaction mixture was stirred at reflux for 5 hours. The precipitate was hot filtered and washed with hot isopropanol (2 L). The resulting white solid was then dried to give DL-3-amino-3- (3-pyridyl) propionic acid as a white solid (220 g).

NMR and MS were consistent with the desired product.

Step 2

The DL-3-amino-3- (3-pyridyl) propionic acid (220 g) from Step 1 was suspended in absolute EtOH (3.6 L). Hydrogen chloride was bubbled through the reaction mixture for 40 minutes while stirring. The suspension was then heated at reflux for 4 hours (the solution was formed after 1 to 1.5 hours). The reaction mixture was cooled to 5 ° C in an ice bath. After stirring at 5 ° C for 1.5 hours, the resulting white precipitate was filtered off and washed thoroughly with ether. After drying under vacuum at 50 ° C, the desired product, ethyl (DL-3-amino-3- (3-pyridyl) propionate) dihydrochloride, was obtained as a white solid (331.3 g).

NMR and MS were consistent with the desired product.

Step 3

To ethyl (DL-3-amino-3- (3-pyridyl) propionate) dihydrochloride (220.6 g, 0.83 mol) from Step 2 in anhydrous THF (2 L) and triethylamine (167.2 g, 1.65 mol) was added portionwise Nt-BOC-glycine N-hydroxysuccinimide ester (225 g, 0.826 mol) (Sigma) at 5 ° C to 10 ° C. The reaction mixture was stirred overnight at room temperature. The resulting precipitate was filtered and washed with THF. The solvent of the filtrate was removed in vacuo. The residue was dissolved in ethyl acetate (2.3 L). The ethyl acetate layer was washed with a saturated solution. 4 4 4 »

9 9 9 9 9

44 49 4 49 444 sodium carbonate (2 × 900 ml) and H ₂ O (3 x 900 ml), dried over MgSO 4 and removed in vacuo. The residue was suspended in 10% ethyl acetate / hexane (2.5 L) overnight. The precipitate was filtered off, washed with 10% ethyl acetate / hexane solution (1 liter), then hexane and dried to give ethyl (3 - [[2 - [[1,1-dimethylethoxy) carbonyl] amino] acetyl] amino] pyridine -3-propanoate) as a white solid (233 g).

NMR and MS were consistent with the desired product.

Step 4

Ethyl 3 - [[2 - [[(1,1-dimethylethoxy) carbonyl] amino] acetyl] amino] pyridine-3-propanoate) (Step 3) (232 g, 0.66 mol) was dissolved in hot dioxane (1 liter). After cooling to room temperature, 4M HCl in dioxane (1.6 L) (Aldrich) was added slowly. After a few minutes a white precipitate formed which was converted into a thick sticky substance. After 2 hours the solvent was decanted. The residue was suspended in ether and the ether was decanted until only a white solid was obtained. This was dried in vacuo to give the bis hydrochloride salt of ethyl 3 - [(2-aminoacetyl) amino] pyridine-3-propanoate) as a white hygroscopic solid (224.2 g).

NMR and MS were consistent with the desired product.

Example K

Preparation of 2-guanidine-4-carboxythiazole hydrochloride

NH

Step 1

To 2-imine-4-thiobiuret (11.1 g, 0.094 mol) (Aldrich) in absolute EtOH (100 mL) was added portionwise ethyl (bromopyruvate) (20.0 g, 0.102 mol) (Aldrich) at reflux. . The solution was stirred at reflux for 2 hours. Additional ethyl bromopyruvate (2.0 g) was added and the reaction 0 0 * 0 0 ·

0 0 0 0 · 0 0000 00

000 0000000 0 • ·· · continued at reflux for a further 2 hours. The reaction was cooled to 10 ° C and concentrated NH ₄ OH was added until pH = 10. The resulting precipitate was filtered, washed with ether and dried to give 2-guanidine-4-carboxyethyl thiazole as a yellow solid (15.1 g) .

MS and ¹ H-NMR were consistent with the desired structure.

Step 2

To the product of Step 1 (5.0 g, 0.23 mol) suspended in H ₂ O (100 mL) and ethanol (40 mL) was added NaOH (0.93 g, 0.023 mol). The solution was stirred overnight at room temperature. Additional ethanol (20 mL) and NaOH (0.93 g) were added to the reaction mixture, and the solution was stirred at room temperature for 1 hour. The pH was lowered to 7 with 1M HCl and the resulting precipitate was filtered, washed with H ₂ O and ether and then dried to give the product (4.1 g) as the zwitterion.

The double ion was suspended in H ₂ O and acidified with concentrated HCl until dissolved. Freezing and lyophilizing the solution gave 2-guanidine-4-carboxythiazole hydrochloride as a pale yellow solid (4.33 g).

MS and ¹ H-NMR were consistent with the desired structure.

Examples L and M were prepared according to the schemes from commercially available 2-amino-6-methylpyridine and 2-amino-4-methylpyridine, respectively.

Example L

Preparation of 2-aminopyridine-6-carboxylic acid

···· 4 · · * · · ·

Step A

To a solution of 2-acetylamine-6-methylpyridine (10.0 g) in water (125 mL) was added portionwise at 75 ° C KMnO ₄ (21.0 g) and heating was continued for another 2 hours. The reaction mixture was cooled, filtered and the residue washed with hot water. (2 x 25 mL). The combined aqueous filtrate and washings were washed with dichloromethane (3 x 40 mL) and acidified with cold 3M HCl. The resulting precipitate was filtered, washed with water to neutral pH and dried in vacuo to give 2-acetylaminopyridine-6-kyric acid (3.8 g, 48% yield). The dichloromethane extract was concentrated to dry residue for and unreacted 2-acetylamine-6-methylpyridine (3.5 g) was obtained.

¹ H-NMR (CD ₃ OD) δ 8.32 (m, 1H), 7.94 (t, 1H), 7.87 (m, 1H), 2.2 (s, 3H); FAB MS m / z 181 (M + H) &lt; + &gt;.

StepB

A suspension of 2-acetylaminopyridine-6-carboxylic acid (4.1 g) in 2.5 M NaOH (36.5 mL) was heated at 80 ° C for 1.5 hours under a nitrogen atmosphere. The resulting solution was cooled and acidified with cold 3M HCl. The precipitate obtained was filtered, washed well with water and acetonitrile. The obtained white solid was dried under vacuum to give 2-aminopyridine-6-carboxylic acid (2.4 g, 76% yield).

¹ H-NMR (DMSO-d ₆ ) δ 7.57 (t, 1H, J = 8.1 Hz), 7.14 (d, 1H, J = 7.2 Hz), 6.67 (1H, J = 8.1 Hz), 6.51 (br, 2H); FAB MS m / z 139 (M + H) &lt; + &gt;.

Step C

A suspension of 2-aminopyridine-6-carboxylic acid (2 g) in EtOH (10.0 mL) and 4M HCl / dioxane (10.0 mL) was heated to reflux for 16 hours under anhydrous conditions. The reaction mixture was then concentrated to a dry residue and dried in a desiccator under vacuum to give 2-amino-6-carboxypyridine hydrochloride as a white powder (1.6 g). ¹ H-NMR (DMSO-d ₆ ) δ 8.3 (br), 7.94 (m, 1H), 7.37 (d, 1H, J = 7.2 Hz), 7.22 (dd, 1H) J = 8.7 Hz), 4.37 (q, 2H, J = 7.3 Hz), 1.32 (t, 3H, J = 7.2 Hz); FAB MS m / z 167 (M + H) &lt; + &gt;.

· Toto to to to to toto toto toto toto toto toto to to to to to to to to to to to to

Example M

Preparation of 2-aminopyridine-4-carbethoxypyridine hydrochloride

Compound (M) was prepared by a procedure similar to the procedure for preparing 2-aminopyridine-6-carbethoxypyridine hydrochloride from Example L from 2-acetylamine-4-methylpyridine starting material.

¹ H-NMR and MS were consistent with the desired structure.

Example 1

Preparation

To the product of Example A (0.5 g, 0.002 mol), the product of Example E (0.2 g, 0.002 mol), triethylamine (0.2 g, 0.002 mol) and DMAP (24 mg) in anhydrous DMA (5 mL) was added EDCI (0.38 g, 0.002 mol) at ice bath temperature. The reaction was stirred at room temperature for 2 days. The ester product was isolated by preparative reverse phase HPLC. To the ester in H ₂ O (3 mL) and CH ₃ CN (3 mL) was added LiOH (0.51 g,

4 ·· ·· • 44 • • · • 4 • · • · • • 4 4 4 4 4 44 • • 4 44 • · · • 4 44 £ 4 4 4 4 4 44 44

0.012 mol). The mixture was stirred at room temperature for 1 hour. The pH was reduced to 2 with TFA and the product was isolated by preparative reverse phase HPLC to give (after lyophilization) the desired product as a white solid (350 mg).

¹ H-NMR and MS were consistent with the desired structure.

Example 2

Preparation

The title compound was prepared according to the procedure in Example 1, substituting an equivalent amount of the product of Example D with the product of Example E, to give the desired product as a white solid (210 mg).

¹ H-NMR and MS were consistent with the desired structure.

Example 3

Preparation

To the product of Example C (19.5 g, 0.045 mol) and N-methylmorpholine (9.1 g, 0.09 mol) in anhydrous DMA (85 mL) was added slowly to a flame-dried flask under nitrogen at ice bath temperature. isobutyl chloroformate (6.2 g, 0.045 mol). The solution to this solution to q-i this # # · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

OJ ««. The mixture was stirred for 15 minutes at ice bath temperature. Then the product of Example D (15 g, 0.04 mol) was added at an ice bath temperature, and then N-methylmorpholine (4.1 g, 0.04 mol) was added slowly at the same temperature. The reaction mixture was stirred overnight at room temperature. To the ester in H ₂ O (50 mL) / CH ₃ CN (30 mL) was added LiOH (16 g, 0.38 mol). The mixture was stirred at room temperature for 1 hour. The pH was reduced to 2 with TFA and the product was isolated by preparative reverse phase HPLC to give (after lyophilization) the desired product as a white solid (13.7 mg).

¹ H-NMR and MS were consistent with the desired structure.

Example 4

Preparation

The above compound was prepared according to the procedure of Example 3, substituting an equivalent amount of the product of Example E with the product of Example D.

¹ H-NMR and MS were consistent with the desired structure.

♦

Example 5

ΓΑ

Preparation

The above compound was prepared according to the procedure of Example 3, substituting an equivalent amount of the product of Example F with the product of Example D.

¹ H-NMR and MS were consistent with the desired structure.

Example 6

Preparation

The above compound was prepared according to the procedure of Example 3, substituting an equivalent amount of the product of Example B with the product of Example C.

¹ H-NMR and MS were consistent with the desired structure.

Example 7

Preparation

OH

To the product of Example 9 (0.6 g, 0.0019 mol) in anhydrous DMA (4 mL) at an ice bath temperature was added isobutyl chloroformate (0.27 g, 0.002 mol) followed by Nmethylmorpholine (0.4 g, 0). , 0038 mol). The solution was stirred at ice bath temperature for 15 minutes. Then the product of Example H (0.71 g, 0.0017 mol) was added at an ice bath temperature followed by N-methylmorpholine (0.17 g, 0.0017 mol). The reaction was stirred overnight at room temperature. The presence of product and significant amounts of starting material were determined by HPLC analysis. EDCI (0.38 g, 0.002 mol) and DMAP (15 mL) were added to the reaction mixture and the reaction was then stirred overnight at room temperature. The ester product was isolated by preparative HPLC. To the ester in H ₂ O (10 mL) and CH ₃ CN (5 mL) was added LiOH (700 mg, 0.017 mol). The mixture was stirred at room temperature for 1 hour. The pH was reduced to 2 with TFA and the product was isolated by preparative reverse phase HPLC to give (after lyophilization) the desired product as a white solid (270 mg).

¹ H-NMR and MS were consistent with the desired structure.

Example 8

Preparation

To the product of Example K (0.5 g, 0.0022 mol) and N-methylmorpholine (0.23 g, 0.0022 mol) in anhydrous DMF (8 mL) was added isobutyl chloroformate (0.31 g) at an ice bath temperature. , 0.0022 mol). After stirring at ice bath temperature for 5 minutes, the product of Example J (0.73 g, 0.0022 mol) and N-methylmorpholine (0.45 g, 0.0045 mol) in anhydrous DMF (8 mL) were added at a single temperature at the same temperature. ). The reaction mixture was stirred at room temperature overnight. The ester was isolated by reverse phase preparative HPLC (530 mg).

To this ester (400 mg) in H ₂ O (10 mL) was added LiOH (91 mg). The mixture was stirred at room temperature for 1 hour. The pH was adjusted to 3 with TFA and the product was isolated by preparative reverse phase HPLC to give (after lyophilization) the desired product as a white solid (350 mg).

¹ H-NMR and MS were consistent with the desired structure.

Guanidation of 2-amino-6-carbethoxypyridine hydrochloride and 2-amino-4-carbethoxypyridine hydrochloride was performed using tris-BOC as shown in Examples N and O (Kim, SK, Qian, L., Tetrahedron Lett. 34, 7677). , 1993) ························

Example N

To a solution of 2-amino-6-carbethoxypyridine hydrochloride (0.56 g, 2.8 mmol) and tris-BOC (91.2 g, 2.8 mmol) in degassed DMF (7.0 mL) was added mercuric chloride (0.76 g, 2.8 mmol) and triethylamine (0.79 g, 7.8 mmol). The resulting mixture was heated at 70 ° C for 24 h under nitrogen and filtered. DMF was distilled under vacuum, the residue was triturated in ethyl acetate and filtered. The orange-colored filtrate was concentrated and the resulting material was purified by silica gel flash chromatography using ethyl acetate containing 1% triethylamine as eluent. Appropriate fractions (blue fluorescent) were combined and concentrated to dryness under reduced pressure to give a yellow powder. MS analysis [m / z 565 (M + H)] confirmed formation of the desired tris-BOC product N.

Example O

O ^ QEt

NH ₂ .HCl

BOC +

BOC

BOC

HgCl ₃ , Et ₃ N c DMF, 70 ° C for 24 h

OF

BOC

••••• 4 · 4 · 4 · 4 · 4 · 4 4 4 4

4,444 «4 4 4

4 4 444444 44 44 44 44 44 44 44 44

This compound (O) was prepared using a procedure similar to Example N. MS analysis [m / z 565 (M + H)] confirmed formation of the desired tris-BOC product O.

The following compounds P and Q can be prepared according to the conditions outlined in step 3 to 5 of Example 9.

Example P

Example R

The above compound was prepared using the procedures described in Example G. In Step 2, 3,5-dichlorosalicylaldehyde was used in place of 3-bromo-5-chlorosalicylaldehyde.

Cl

T o γ0γ ° ^{η HCl}

Br

999999 9 9 9 ·

9 9 9 9 9 9 9 9999999 9 9 9 9

Scheme of Example 9

O ······························ 9 4 ·· 9 · 9

Example 9 (scheme shown above)

Step 1

To a mechanically stirred suspension of Reagent 1 (18.68 g) in THF (11) at 0 ° C was added sodium hydride (19 g of 60% suspension in mineral oil) at 0 ° C over 30 minutes. After 30 minutes, pure di-tec was added. butyldicarbonate (95 g) and the reaction mixture was gently heated at reflux for 16 h. The mixture was cooled to 0 ° C and quenched with saturated NaHCO 3 solution. The mixture was extracted with ethyl acetate. The organic extract was washed with water, dried (Na ₂ SO ₄ ) and concentrated. The residue was flash chromatographed using a small column packed with silica gel (60 mm x 180 mm) to give the desired product 2 as a thick yellow liquid (68 g). NMR was consistent with the desired structure.

Step 2

A stirred suspension of 3-amino-pyridine-6-carboxylic acid (25 g) in ethanol (300 mL) was saturated with hydrogen chloride gas at 0 ° C. The mixture was heated to 23 ° C and heated at reflux for 2 hours. A clear solution was obtained. After cooling to 23 ° C, the mixture was concentrated in vacuo, neutralized with aqueous NaHCO ₃ and extracted with ethyl acetate. The organic phase was washed with water, dried over MgSO ₄ and concentrated in vacuo to give ethyl (3-amino-pyridine-6-carboxylate) (25.0 g), triethylamine (27 mL), ethyl 3-amino-pyridine-6-carboxylate (13 g) and DMF (200 mL) were stirred and heated at 55 ° C for 30 hours. The reaction mixture was cooled to 23 ° C, diluted with ethyl acetate (500 mL) and filtered through celite. The filtrate was washed with water (2x), dried over MgSO ₄ and concentrated under vacuum. The residue was chromatographed on silica gel using ethyl acetate in hexane as eluent to give the desired intermediate 3. NMR was consistent with the desired structure.

Step 3

A suspension of the product of Step 2 (13 g) in 5M hydrochloric acid (100 mL) was stirred at 23 ° C for 16 hours. The mixture was concentrated in vacuo. The residue was suspended with methanol and the white solid was filtered to give the desired product

4. NMR and MS were consistent with the desired structure.

999999 · »

999 9 99 · ··· 99 9999 99 9

999 9999999 9 9

9900 99 99 99 9

99 99 tt. «9 999

Step 4

The product of Step 3 (0.308 g) was suspended in DMF (10 mL) and 4-methylamorpholine (0.2 mL) was added. The mixture was stirred at 23 ° C for 1 hour. After cooling the reaction mixture to 0 ° C, IBCF (0.129 mL) was added. After half an hour, a solution of the product of Example R (0.416 g) and 4-methylmorpholine (0.11 mL) in DMF (3 mL) was added. The mixture was heated at 23 ° C for a further 2 hours. Then the mixture was filtered and the filtrate was concentrated in vacuo. Purification of the residue by HPLC gave product 5. Micro-analytical results, NMR and MS were consistent with the desired structure.

Step 5

A solution of the product of Step 4 in 3.5M HCl was allowed to stand at 23 ° C for 3 hours, concentrated in vacuo, and purified by HPLC to afford the desired product. The microanalytical results, NMR and MS were consistent with the desired product.

Analysis calculated for C ₂₁ H ₂₂ BrClN ₆ O ₆ . 2CF ₃ CO ₂ H. H ₂ O:

C, 36.80; H, 3.21; N, 10.30

Actual values; C, 36.52; H, 2.90; N, 9.91.

Example 10 »44 4 · 444 9 4 • 44 4 4 9 9 4

4 9 4444 4 4 • 444 4444 4444 44 44 4 4

HO ', ΝΗ

TFA

The compound was prepared using the procedure described in Example 9, except that the product of Example G was used in place of the product of Example R, Step 4. Step.

Analysis calculated for C ₂₁ H ₂₂ Cl ₂ N ₆ O ₆ . 2CF3CO2H. H ₂ O:

C, 38.93; H, 3.40; N, 10.89;

Found: C, 39.27; H, 3.12; N, 11.09.

Example 11

The compound was prepared as described in Example 9 using the product of Example I instead of the product of Example R in Step 4.

Analysis calculated for C ₂₁ H ₂₁ Br ₂ N ₆ O 6 · 2 CF ₃ COOH H ₂ O:

C, 35.65; H, 2.87; N, 9.98

Found: C, 35.81; H, 2.79; N, 10,14 • fefe fefe fefe fefe fefe fefefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe fefe · ·· fefe · · fefe ·

Example 12

Preparation of (3S) -N - [[4 - [(1,4,5,6-tetrahydro-5-hydroxy-2-pyrimidinyl) -amino] -2-thienyl] carbonyl] glycyl-3- (3, trifluoroacetate salt) 5-dichloro-2-hydroxyphenyl) -p-alanine

MeOH,

INNaOH

Step 1

A solution of 5-bromo-2-thiophene carboxylic acid (5.0 g) in concentrated sulfuric acid (30 mL) was cooled to -30 ° C and treated dropwise with a solution of 70% nitric acid (7.7 mL) (d). = 1.40) in concentrated sulfuric acid (30 mL). The reaction mixture was stirred at -25 ° C for 30 minutes and then poured into ice-water slurry and stirred for several minutes. The precipitated substance was filtered, recrystallized from water and dried to give a white powder (2.3 g). ¹ H-NMR was consistent with the desired structure.

• φ φφ φ • φ ·

Step 2:

A solution of the product of Step 1 (1.7 g) in THF (25 mL) was treated with a catalytic amount of 5% Pd / C under a hydrogen atmosphere at 50 psi for 16 hours at room temperature. The reaction mixture was filtered and concentrated. The residue was heated with ethyl acetate and filtered to give a brown solid (750 mg). ¹ H-NMR was consistent with the desired structure.

Step 3

A solution of the product of Step 2 (725 mg) with guanylating agent G1 in THF (15 mL) and dimethylacetamide (10 mL) was refluxed for 16 hours. The reaction mixture was concentrated and the residue was treated with a solution of TFA (5 mL) and CH ₂ Cl ₂ (5 mL). After stirring at room temperature for 1 h, the reaction mixture was concentrated and the residue was purified by reverse phase HPLC using water (0.5% TFA) and acetonitrile gradient as eluent to give a pale yellow thick liquid (300 mg). ¹ H-NMR was consistent with the desired structure.

Step 4

In a flame-dried flask, the product of Step 3 (275 mg) was dissolved in 8 ml of dimethylacetamide and N-methylmorpholine (81 mg) under a nitrogen atmosphere. The stirred solution was cooled to 0 ° C and then added dropwise to a solution of isobutyl chloroformate (109 mg) in dimethylacetamide (1 mL). The reaction mixture was stirred at 0 ° C for 45 min and then treated dropwise with a solution of the product of Example G (286 mg) and N-methylmorpholine (81 mg) in dimethylacetamide (2 mL). The reaction mixture was warmed to room temperature and stirring was continued for a further 16 hours. The reaction mixture was concentrated and the residue was purified by reverse phase HPLC as described in step 3 to give a thick colorless liquid (262 mg). ¹ H-NMR was consistent with the desired structure.

Step 5:

A solution of the product of Step 4 (250 mg) in methanol (8 mL) and 1M sodium hydroxide solution (8 mL) was stirred at room temperature for 16 hours. The reaction was quenched with TFA (0.7 mL) and concentrated.

·········· · · 99 · ·· ··· · 9 9 ·> ·

999 · «· 9 · 9 · 9 9999 · 9 9

The residue was purified by reverse phase HPLC as described in Step 3 to give a white solid (144 mg). ¹ H-NMR was consistent with the desired structure. Analysis calculated for Ca 10 H 11 BrN 5 O 6 Cl 2 S. 1.50 TFA. 0,25 H ₂ O:

C, 39.13; H, 3.28; N, 9.92; S, 4.54; Cl, 10.04;

Found: C, 38.81; H, 3.17; N, 9.90; S, 4.86; Cl, 10.25.

Example 13

Preparation of N - [[4 - [(4,5-dihydro-1H-imidazol-2-yl) amino] -2-thienyl] carbonyl] glycyl-3- (3,5-dichloro-2-hydroxyphenyl) trifluoroacetate salt - p-alanine

The above compound was prepared by a similar sequence of reactions used to prepare the compound described in Example 12. In Step 3 of Example 12, the guanylating reagent described in Example C (Step 1) was used instead of G1. Similarly, the product of Example D was used in place of the product of Example G, in Step 4.

¹ H-NMR was consistent with the desired structure.

Analysis calculated for CwHwhfeO5CfeS. 1.50 TFA. 0.50 H ₂ O:

C, 38.84; H, 3.18; N, 10.29; S, 4.71; Cl, 10.42;

Found: C, 38.84; H, 3.01; N, 10.50; S, 5.08; Cl, 11.01.

••• to · to · to · to · to · to ···

this · · · · · this · · ··· «• to · ·· to

Example 14

Preparation of (3S) -N - [[5-methyl-4 - [(1,4,5,6-tetrahydro-5-hydroxy-2-pyrimidinyl) amino] -2-thienyl] carbonyl] glycyl-3- trifluoroacetate salt (3,5-dichloro-2-hydroxyphenyl) -β-alanine

Step 1

Compound (A) was prepared as described in Step 1 of Example 13 using 5-methyl-2-thiophene carboxylic acid (5.2 g). The product was purified by extraction with dilute sodium hydroxide solution, dilute acidification with dilute hydrochloric acid and ex97.

4,494 4 4 4

4 4 4 4

4 4 44

4 4

4 9

4 4 4

4 4 4

44 by traction into ethyl acetate. The organic extract was dried (Na ₂ SO ₄ ) and concentrated to give a yellow-orange solid (2.2 g). ¹ H-NMR was consistent with the desired structure.

Step 2

Compound (B) was prepared using the product of Step 1 (2.2g) and under the coupling conditions described in Step 4 of Example 12. The reaction was concentrated and the residue was partitioned between ethyl acetate and water. The organic extract was washed with saturated sodium hydroxide solution, dried (Na ₂ SO ₄ ) and concentrated. The residue was recrystallized from ethyl acetate to give a tan solid (3.5 g). ¹ H-NMR was consistent with the desired structure.

Step 3

A solution of step 2 product (2.8 g) in THF (25 mL) was added dropwise to a solution of stannous chloride dihydrate in concentrated hydrochloric acid (1.0 g / 2 mL) at room temperature until TLC showed complete disappearance of the starting material. The organic solvent was removed and the aqueous portion was neutralized with sodium carbonate solution. The aqueous portion was extracted with CH ₂ Cl ₂ , dried (Na ₂ SO ₄ ), and concentrated to give a yellow powder (1.1 g). ¹ H-NMR was consistent with the desired structure.

Step 4

To a solution of the product described in Step 3 (375 mg) and the product from Step 1 of Example 9 (519 mg) in DMF (15 mL) was added mercuric chloride (326 mg) and triethylamine (182 mg). The mixture was heated at 95 ° C to 100 ° C for 3 hours. The reaction mixture was cooled, mixed with ethyl acetate (30 mL), stirred for 30 minutes and then filtered through a celite pad. The filtrate was concentrated and purification of the residue by flash chromatography (eluting with 97.5% CHCl ₃ - 2.5% CH ₃ OH) gave a tan solid (415 mg). ¹ H-NMR was consistent with the desired structure.

Step 5

A solution of the product of Step 4 (400 mg) in CH ₂ Cl ₂ (7.5 mL) and TFA (7.5 mL) was stirred for 1 hour at room temperature. The reaction was concentrated and the residue was stirred for 16 hours at room temperature in a solution of CH ₃ OH (10 mL) and 1M sodium hydroxide solution (10 mL). The reaction was quenched with THF (0.8 mL) and concentrated. The residue was purified using reverse phase HPLC as above to give a white solid (80 mg). ¹ H-NMR was consistent with the desired structure.

Calcd for C21H23N5O6Cl2.1.50 TFA. 0.50 H ₂ O:

C, 39.79; H, 3.55; N, 9.67; S, 4.43; Cl, 9.79;

Found: C, 39.50; H, 3.24; N, 9.58; S, 4.71; Cl, 10.22.

Example 15

Preparation of (3S) -N - [[4- (4,5-dihydro-1H-imidazol-2-yl) amino] -5-methyl-2-thienyl] carbonyl] glycyl-3- (3,5-dichloro- 2-hydroxyphenyl) -p-alanine

The above compound was prepared by a similar sequence of reactions used to prepare the compound described in Example 14, except that the guanylating reagent described in Example C (Step 1) was used instead of the reagent used in Step 4 of Example 14. The HNMR was consistent with the desired structure.

Calcd for C20H21N2O5Cl2S. 1.50 TFA. 0,25 H ₂ O:

C, 40.04; H, 3.36; N, 10.15; S, 4.65; Cl, 10.28;

Found: C, 39.82; H, 3.19; N, 10.17; S, 4.86; Cl, 10.69.

• · · · · ·

Example 16

N - [[5 - [(4,5-dihydro-1H-imidazol-2-yl) amine] -2-thienyl] carbonyl] glycyl trifluoroacetate salt

3- (3,5-dichloro-2-hydroxyphenyl) -3-alanine fcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcfcf «· · Fcfcfcfc · fcfc · fcfc · fcfc · · · fc ··

z fM

100 ···· ΦΦ · · φ • • φ «» »» »· · · · · · · · · ·

The title compound was synthesized from the starting 2-nitro-5-thiophenecarboxylic acid using the same sequence of reactions as described in Example 15. The conditions described in Step G, Example 17 were used for the guanylation reaction.

Calcd for C19HWN2O5Cl2S. 1.50 TFA:

C, 39.36; H, 3.08; N, 10.43; S, 4.78;

Found: C, 39.05; H, 2.79; N, 10.37; S, 4.90.

Example 17

N - [[5 - [(4,5-dihydro-1H-imidazol-2-yl) amino] -6-methoxy-3-pyridinyl] carbonyl] glycyl-3- (3,5- dichloro-2-hydroxyphenyl) -3-alanine

• 0 * · 00

0 0 0 0 0 ιοί. :::

«0 00

Step 1

A mixture of 6-hydroxynicotinic acid (20 g) and colorless fuming nitric acid (60 mL) (d = 1.50) was heated at 90-95 ° C for 3 hours. The reaction was cooled to room temperature and poured into 1.5 liters of ice-water slurry with stirring and filtered after 15 minutes. The precipitate was washed with water and dried to give a pale yellow powder (9.3 g).

¹ H-NMR was consistent with the desired structure.

Analysis calculated for C ₆ H ₄ N ₂ O ₅ .

C, 39.14; H, 2.19; N, 15.22.

Found: C, 39.08; H, 2.17; N, 15.19.

Step 2

A mixture of the product of Step 1 (5.0 g) and phosphorus oxychloride (15 mL) was heated to reflux for 3 hours. The reaction mixture was cooled to room temperature, added to ice-water and stirred for 30 minutes. Additional ice was added to keep the mixture cooled during this time. The reaction mixture was extracted with THF-ether (1: 2), the organic extracts were washed with saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated. The residue was recrystallized from ether: hexane 1: 1 to give a yellow powder (4.0 g).

¹ H-NMR was consistent with the desired structure.

Step 3

A solution of the product of Step 2 (1.6 g) in thionyl chloride (5.0 mL) was heated at reflux for 3 hours. The reaction was cooled to room temperature and evaporated under a stream of nitrogen. The residue was dried under vacuum and used without further purification. In a flame-dried vessel under nitrogen, the product of Example D (3.0 g) was dissolved in dimethylacetamide (30 mL) and Ν, Ν-diisopropylethylamine (2.3 g). The solution was cooled in an ice bath, and a solution of hydrochloric acid (obtained above) in THF (20 mL) was added dropwise. The reaction was stirred, warmed to room temperature and then quenched with water (25 mL). The mixture was extracted with ethyl acetate and the organic extracts were washed with saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated. The resulting yellow solid was recrystallized from ethyl acetate to give a pale yellow powder (3.4 g). ¹ H-NMR was consistent with the desired structure.

102 «444 4 ·« 4 4 · «4 β · *« 34 »···» 44 4 · 4 · 44 · «4 4 ·« 4 4 «44 4 4 4 4

4444 «4 4 44 4 • 4 4» 44 4 44 44 «

Step 4

Κ a suspension of sodium methoxide (2.2 g) in methanol (30 ml) was added dropwise at 0 ° C a solution of the product of step 3 (4.5 g) in methanol (30 ml). The reaction was stirred for 30 minutes and then quenched with acetic acid (2.3 mL). The reaction was concentrated and partitioned between ethyl acetate and water. The organic extracts were dried (Na ₂ SO ₄ ), concentrated and purified on a silica gel column (80% ethyl acetate - 20% hexane) to give a pale yellow solid (3.5 g).

¹ H-NMR was consistent with the desired structure.

Step 5

A solution of the product described in Step 4 (1.07 g) in ethanol (25 mL) was treated with a catalytic amount of 3% Pt / C, digested with sulfur at a hydrogen pressure of 34.5 kPa, for 4 hours at room temperature. The reaction mixture was filtered and concentrated to give a white solid (930 mg) which was used without further purification. ¹ H-NMR was consistent with the desired structure.

Step 6

To a solution of Step 5 product (330 mg), G ₂ (276 mg) in DMF (10 mL) was added triethylamine (150 mg) and mercuric chloride (258 mg) and the mixture was heated at 85 ° C for 16 hours. The reaction was cooled, mixed with ethyl acetate (25 mL), and filtered. The filtrate was concentrated and purification of the residue on a silica gel column eluting with 98% CH ₂ Cl ₂ - 2% methanol gave a white solid (325 mg). ¹ H-NMR was consistent with the desired structure.

Step 7

A solution of the product of Step 6 (114 mg) with TFA (8 mL) and dichloromethane (8 mL) was stirred at room temperature for 90 minutes. The solvent was evaporated and the residue was treated with a solution of 1M sodium hydroxide (8 ml) and methanol (8 ml) for 16 hours at room temperature. The reaction was quenched with TFA (1 mL) and concentrated. The residue was purified using reverse phase HPLC as described above to give a white solid (76 mg). ¹ H-NMR was consistent with the desired structure.

Analysis calculated for C ₂₁ H ₂₂ N ₆ O ₆ Cl ₂ .2 TFA:

• 9 · 9 · · · · · · ·

103. ::

C, 41.74; Η, 3.88; N, 12.70. Found: C, 41.47; H, 3.49; N, 12.85.

Example 18

N - [[5 - [(4,5-dihydro-1H-imidazol-2-yl) amine] -1,6-dihydro-1-methyl-6-oxo-3-pyridinyl] carbonyl] glycyl- trifluoroacetate salt 3- (3,5-dichloro-2-hydroxyphenyl) -β-alanine

Step 1

A suspension of the product of Example 17, Step 1 (12.0 g) in methanol (200 mL) was mixed with concentrated sulfuric acid (1 mL) and heated to reflux. After 16 hours at reflux, the mixture became a homogeneous solution and cooled to room temperature. About half of the solvent was removed and then the product crystallized from solution. The mixture was cooled in an ice bath, filtered and dried to give a yellow powder (16.7 g).

¹ H-NMR was consistent with the desired structure.

Step 2

A solution of the product of Step 1 (2.6 g) in DMF (40 mL) was cooled to 0 ° C under a nitrogen atmosphere and mixed with a 60% dispersion of sodium hydride in mineral oil (655 mg). The mixture was stirred at 0 ° C for 45 minutes and then mixed with iodomethane (2.8 g) at once. The reaction was stirred at room temperature for 16 hours and then partitioned between ethyl acetate and water. The aqueous component was extracted several times with an additional amount of ethyl acetate, and then the combined organic extracts were washed with brine, dried (Na ₂ SO ₄ ) and concentrated. The residue was purified on a silica gel column (eluting with 2% methanol - 98% dichloromethane) to give a yellow solid (1.9 g). ¹ H-NMR was consistent with the desired structure.

Step 3

A solution of the product of Step 2 (2.6 g), methanol (60 ml) and 2M sodium hydroxide solution (60 ml) was stirred at room temperature for 16 hours. The reaction was quenched with glacial acetic acid (6.9 mL) and concentrated. The residue was dried under high vacuum. This material was then refluxed with thionyl chloride (100 mL) for 3 hours under a nitrogen atmosphere. The reaction was then cooled, concentrated and thoroughly dried under high vacuum to give a tan solid (2.2 g). ¹ H-NMR was consistent with the desired structure. In a flame-dried vessel, the product of Example D (3.5 g) was dissolved in Ν, Ν-dimethylacetamide (35 mL) and diisopropylamine (2.9 mL) under a nitrogen atmosphere. The solution was cooled to 0 ° C and treated with a solution of the acid chloride (1.9 g) (obtained previously) in THF (35 mL). The reaction was stirred for 30 minutes and then concentrated under high vacuum to remove the solvent. The residue was partitioned between ethyl acetate and water and the aqueous portion was extracted several times with additional ethyl acetate. The combined organic extracts were washed several times with water and then washed with saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated to give a yellow solid (1.3 g). ¹ H-NMR was consistent with the desired structure.

······ ··· «· • · · · · · ···, _Λ - · · · · · · ···

105 .......... · ♦ · · ·

Step 4

A solution of the product of Step 3 (1.0 g) in THF (10 mL) and DMF (10 mL) was treated with a catalytic amount of 5% Pt / C at a hydrogen pressure of 5 psi for 4 hours at room temperature. The reaction mixture was filtered and concentrated. The residue was mixed with a minimum amount of THF and slowly crystallized at room temperature. An equivalent volume of ethyl acetate was added to the crystallized mixture, which was left for 15 minutes. The mixture was cooled, filtered and washed with cold ethyl acetate. Drying of the product gave a white solid (730 mg). ¹ H-NMR was consistent with the desired structure.

Step 5

To a solution of the product of Step 4 (720 mg) and G ₂ (581 mg) in DMF (25 mL) was added triethylamine (304 mg) and mercuric chloride (544 mg). The mixture was heated at 85 ° C under nitrogen for 1 hour. The reaction mixture was cooled and filtered through celite. The filtrate was concentrated and purified on a silica gel column (10% methanol - 90% chloroform) to give a white solid (375 mg). ¹ H-NMR was consistent with the desired structure.

Step 6

The above compound was prepared from the compound prepared in Step 5 (360 mg) using the procedure described in Example 14 (Step 5). The crude product was purified using reverse phase HPLC as above to give a white solid (223 mg). ¹ H-NMR was consistent with the desired structure.

Calcd for C21H22N6O6Cl2.5 TFA. 0.5 H ₂ O:

C, 38.11; H, 3.14; N, 10.26; Cl, 8.65.

Found: C, 38.18; H, 3.05; N, 10.79; Cl, 9.34.

Example 19

N - [[6-chloro-5 - [(4,5-dihydro-1H-imidazol-2-yl) amino] -3-pyridinyl] carbonyl] glycyl-3- (3,5-dichloro- 2-hydroxyphenyl) -p-alanine

106

Step 1

A solution of stannous chloride (6.3 g) in concentrated hydrochloric acid (10 ml) was prepared and added dropwise to a solution of the product of Step 3, Example 17 (1.0 g) in THF (15 ml). The reaction was stirred for 15 minutes and then cooled to room temperature. The organic solvent was removed to give a gummy consistency. The aqueous component was decanted and the gummy precipitate was partitioned between ethyl acetate and dilute sodium bicarbonate solution. The aqueous component was extracted several times with additional ethyl acetate and the combined organic extracts were then washed with water and saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated. The residue was dried under vacuum to give a tan powder (375 mg). ¹ H-NMR was consistent with the desired structure.

Step 2

The compound described in Step 1 (365 mg) was guanylated according to the procedure used in Step 5 of Example 18. The crude product was purified on a silica gel column (eluting with 2% methanol - 98% chloroform) to give a brownish thick liquid (205 mg). ).

¹ H-NMR was consistent with the desired structure.

Step 3

The above compound was prepared using the product of Step 2 (200 mg) followed by the procedure described in Step 7, Example 17. The crude product was purified using reverse phase HPLC as described above to give a white solid (53mg).

¹ H-NMR was consistent with the desired structure.

107

Analysis calculated for C 20 H 19 N 6 O 5 Cl 3. 1.7 TFA. 0.5 H ₂ O:

C, 37.90; H, 3.09; N, 11.33; Cl, 14.34.

Found: C, 37.53; H, 2.71; N, 11.33; Cl, 15.01.

Example 20

N - [[5 - [(4,5-dihydro-1H-imidazol-2-yl) amino] -1,6-dihydro-6-oxo-3-pyridinyl] carbonyl] glycyl-3- (3) trifluoroacetate salt 5-dichloro-2-hydroxyphenyl) -3-alanine

The above compound was prepared by dissolving the product of Example 17, Step 6 (353 mg) in a 6M solution of hydrochloric acid (50 mL) at room temperature for 48 hours. The reaction was concentrated and purification of the residue by reverse phase HPLC as described previously gave a white solid (115 mg). ¹ H-NMR was consistent with the desired structure.

Analysis calculated for C ₂ oH ₂ O ₆ Cl ON6 _second 1.25 TFA:

C, 41.33; H, 3.28; N, 12.85.

Found: C, 41.57; H, 3.29; N, 12.92.

Example 21

Preparation of (3S) -N - [[1,6-dihydro-6-oxo-5 - [(1,4,5,6-tetrahydro-5-hydroxy-2-pyrimidinyl) amino] -3-pyridinyl] trifluoroacetate salt carbonyl] glycyl-3- (3,5-dichloro-2-hydroxyphenyl) -β-alanine

108

The starting material used in the preparation of said compound was prepared according to the sequence of reactions described in Example 17 (steps 1 to 3). Instead of the product of Example D, the chiral intermediate of Example G was used in the coupling reaction in Step 3.

······ · · · · · · ·

Step 1

Compound (A) was prepared according to the procedure of Step 4, Example 17.

¹ H-NMR was consistent with the desired structure.

109 • · · · · ····· + · ·

Step 2

A solution of the compound of Step 1 (2.7 g) in 4M anhydrous HCl / dioxane (100 mL) was heated at 45 ° C under nitrogen for 72 h. Concentration to dryness gave a light brown solid (2.6 g).

¹ H-NMR was consistent with the desired structure.

Step 3

A solution of the product of Step 2 (1.85 g) and ethanol (50 mL) was reacted with a catalytic amount of 5% Pt / C under a hydrogen pressure of 34.5 kPa for 2 hours at room temperature. The reaction mixture was filtered and concentrated and the residue was purified by reverse phase HPLC to give a white solid (507 mg). ¹ H-NMR was consistent with the desired structure.

Step 4

To a solution of Step 3 product (500 mg) and Step 1 product of Example 4 (650 mg) in DMF (15 mL) were added triethylamine (304 mg) and mercuric chloride (408 mg) and the mixture was heated at 100 ° C under nitrogen. The reaction mixture was cooled to room temperature, stirred with ethyl acetate for 15 minutes and then filtered. The filtrate was concentrated and the residue was treated with a solution of TFA (7 mL) and CH ₂ Cl ₂ (7 mL) at room temperature for 1 hour. The mixture was concentrated, and the residue was purified by reverse phase HPLC as described above to give a white solid (400 mg).

¹ H-NMR was consistent with the desired structure.

Step 5:

A solution of the product of Step 4 (200 mg) was stirred for 16 hours at room temperature with 1M sodium hydroxide (8 ml) and methanol (8 ml). The reaction was quenched with TFA (1 mL) and concentrated. The residue was purified by reverse phase HPLC as described above to give a white solid (79 mg). ¹ H-NMR was consistent with the desired structure.

······ · · · · · · · · · · · · · · · · · · ·

110 • · ··· ······· · · · · ···

Analysis calculated for C 21 H 22 N 6 O 7 Cl 2. 1,5 TFA:

C, 40.46; H, 3.32; N, 11.80.

Found: C, 40.12; H, 3.57; N, 12.26.

Example 22

Preparation of N - [[5 - [(4,5-dihydro-1H-imidazol-2-yl) amino] -2-methoxy-3-pyridinyl] carbonyl] glycyl-3- (3,5-dichloro-2) trifluoroacetate salt -hydroxyfernyl) -3-alanine.

X γϊ τ

O

111 9 9 9 9 99 9 9 9 9 9 9 9 9 9 9 99 9 9 9 9 9 9 9 9 9 9 99 9 ··

Step 1

To a solution of 2-hydroxynicotinic acid (10.0 g, 71.9 mmol) in concentrated sulfuric acid (28.6 mL) was added fuming nitric acid (7.1 mL) dropwise. The reaction mixture was heated at 55 ° C for 4 hours. The cooled reaction mixture was poured into ice water. The product precipitated as a yellow solid. The precipitated solid was removed by filtration, washed with water and air dried to give the desired product (7.2 g, 54% yield). NMR was consistent with the desired structure.

Step 2:

A solution of the product of Step 1 (5.0 g, 27.2 mmol) in phosphorus oxychloride (13.5 g) was refluxed for 4.5 hours. The cooled reaction mixture was poured into ice water (200 g). The resulting mixture was stirred for 30 minutes and extracted with tetrahydrofuran / ethyl acetate (2/1). The organic layer was washed with brine, dried over Na ₂ SO ₄ and filtered. Concentration of the filtrate gave an oil. Mixing the oil with hexane / ether (4/1) gave the desired product as a bright yellow powder (5.0 g, 91% yield). NMR was consistent with the desired structure.

Step 3

A solution of the product of Step 2 (3.1 g, 15.3 mmol) in thionyl chloride (8.1 mL) was refluxed for 4.5 hours. The residual thionyl chloride was removed from the reaction mixture under reduced pressure to give a light brown oil. Drying the oil under vacuum gave the product as a yellow solid (2.7 g, 79% yield). NMR was consistent with the desired structure.

Step 4

To a solution of the product of Example D (5.5 g, 14.8 mmol) in N, N-dimethylacetamide (40 mL) and tetrahydrofuran (15 mL) was added diisopropylethylamine (3.7 g, 4.9 mL) and the reaction mixture. was cooled to -5 ° C. A solution of the product of Step 3 (3.1 g, 14.1 mmol) in tetrahydrofuran (25 mL) was added to the reaction mixture over 15 minutes. The reaction mixture was stirred at -5 ° C for 30 minutes and then warmed to room temperature. After 3 hours, tetrahydrofuran was removed from the reaction mixture under reduced pressure. The reaction mixture was poured into ice water. The precipitated solid was collected by filtration, washed with water and air dried to give the product as a yellow solid (6.2 g, 85% yield). NMR was consistent with the desired structure.

112

4 4 4 4 4 4 4 4 4 4

44444

4 4

4

Step 5:

A solution of the product of Step 4 (4.0 g, 7.7 mmol) was stirred at 0 ° C with sodium methoxide (1.67 g, 30.8 mmol) in methanol (45 mL) for 4 h. The reaction was neutralized with acetic acid. Methanol was removed from the reaction mixture and purification of the crude product by chromatography (silica gel, ethyl acetate-toluene, 6/4) gave the desired product as a yellow solid (2.5 g, 67% yield). NMR was consistent with the desired structure.

Step 6

A mixture of the product of Step 5 (0.99 g, 1.9 mmol), 3% platinum on carbon and methanol was treated with H ₂ at 50 psi at room temperature for 16 hours. The platinum catalyst was removed by filtration. Methanol was removed under reduced pressure to give the desired product as a yellow solid (0.62 g, 70% yield). This product was used in the next reaction without further purification. NMR was consistent with the desired structure.

Step 7

A mixture of the product of Step 6 (0.26 g, 0.54 mmol), G ₂ (0.18 g, 0.59 mmol), triethylamine (0.25 g, 1.78 mmol), mercuric chloride (0.15 g, 0.25 mmol), 16 g, 0.59 mmol) and N, N-dimethylaformamide (15 mL) were heated at 90 ° C to 95 ° C for 16 hours. The cooled reaction mixture was filtered through a short clone filled with celite with ethyl acetate (40 mL) and then dichloromethane (30 mL). Chromatography of the crude mixture (silica gel, CH ₂ Cl ₂ -CH ₃ OH-NH 4 OH, 95/5 / 0.5) gave the product as a yellow solid (0.31 g, 76% yield). NMR was consistent with the desired structure.

Step 8

A solution of the product of Step 7 (0.31 g, 0.41 mmol) was stirred with trifluoroacetic acid (3.5 mL) and dichloromethane (7.0 mL) at room temperature for 2 hours. Trifluoroacetic acid and dichloromethane were removed under reduced pressure and the crude mixture was treated at room temperature with methanol (5 mL) and dodium hydroxide solution (2M, 2.5 mL) at room temperature for 18 h. Sodium hydroxide was neutralized by the addition of acetic acid. Removal of the methanol gave a crude mixture. The crude mixture was purified using

113

Reversed phase HPLC gave the title compound as a white solid (0.10 g, 36% yield).

Analysis calculated for C 21 H 22 N 6 O 6 Cl 2. 1,3 CF3COOH. 0.25H ₂ O:

C, 41.80; H, 3.54; N, 12.39, Cl, 10.46;

Found: C, 42.12; H, 3.84; N, 11.87; Cl, 10.99.

Example 23

Preparation of N - [[5- (4,5-dihydro-1H-imidazol-2-yl) amino] -1,2-dihydro-2-oxo-3-pyridinyl] carbonyl] glycyl-3- (3) trifluoroacetate salt 5-dichloro-2-hydroxyphenyl) -β-alanine

A solution of the product of Step 7 (0.21 g, 0.28 mmol), Example 22, in dioxane (4 mL) was treated with 6M HCl (6 mL) at room temperature for 72 hours. The solvent was removed under reduced pressure and purification of the crude mixture by reverse phase HPLC using acetonitrile-water as eluent gave the title compound as an off-white solid (0.052 g, 27% yield).

Analysis calculated for C 20 H 20 N 6 O 6 Cl 2. 1.6 CF ₃ COOH:

C, 40.17; H, 3.14; N, 12.11;

Found: C, 40.10; H, 3.22; N, 11.84;

Example 24 ·········

114 9 9

9 9 9

9 9 999

Preparation of N - [[5 - [(4,5-dihydro-1H-imidazol-2-yl) amino] -1-methyl-1H-pyrazol-3-yl] carbonyl] glycyl-3- (3,5-dichloro- 2-hydroxyphenyl) -p-alanine

··· * ·· ·· ·

The first and second steps of the synthesis of the target compound were carried out according to literature procedures (Justus Liebigs. Ann. Chem. 512, 97, 1934 and J. Heterocyclic Chem. 21, 737, 1994, respectively).

Step 3

To a solution of the product of Step 2 (0.9 g, 5.3 mmol) and G ₂ (1.9 g, 5.9 mmol) in DMF (25 mL) was added triethylamine (2.5 mL, 17 mL), 6 mmol) and mercuric chloride (1.6 g, 5.9 mmol). After cooling to 80 ° C to 85 ° C for 18 hours, the reaction mixture was cooled and filtered through a pad of celite. The residue was washed with dichloromethane and the combined fractions were concentrated. The residue was dissolved in dichloromethane and washed with water and brine. After drying over sodium sulfate, the mixture was filtered and concentrated. The crude orange solid (1.8 g) was purified by silica gel chromatography (hexane / ethyl acetate 1/1) to give the N-Boc protected product as an off-white solid white solid (1.4 g).

Step 4

To a solution of the product of Step 3 (2.4 g, 5.5 mmol) in ethanol (50 mL) was added aqueous lithium hydroxide (526 mg in 5 mL of water). The reaction mixture was stirred at room temperature for 24 hours. Acetic acid (22 mL) was added to the reaction mixture and stirred for 15 minutes. After removal of the solvent under reduced pressure, the crude mixture was purified by reverse phase HPLC (acetonitrile / water containing trifluoroacetic acid) to give the product as a TFA salt as a thick white liquid. The residue was mixed with hydrochloric acid and (4M solution in dioxane) and stirred for 10 minutes. The reaction was concentrated to remove the solvent and repeated the process twice to give the desired product as a white solid (3.9 g). The product was used in the next step without further purification.

Step 5:

To a solution of the product of Step 4 (0.9 g, 3.7 mmol) in DMF (15 mL) at 0 ° C was added N-methylmorpholine (605 mL, 5.5 mmol) followed by isobutyl chloroformate (475 mL, 3 mL). , 7 mmol). After stirring for 5 minutes, a solution of the product of Step D in DMF (10 mL) containing N-methylmorpholine (605 mL, 5.5 mmol) was injected into the reaction. The mixture was then stirred at 0 ° C for 1 hour and at room temperature for 36 hours. The solvent was removed after

Purification of the residue by reverse phase HPLC (acetonitrile / water containing trifluoroacetic acid) gave the product (525 mg) as a TFA salt.

Step 6

To a solution of the product of Step 5 (540 mg, 1 mmol) in ethanol (50 mL) was added aqueous lithium hydroxide (144 mg in 10 mL of water). The reaction mixture was stirred at room temperature for 24 hours. Acetic acid (305 mL) was then added to the reaction mixture and stirred for 15 minutes. After removal of the solvent under reduced pressure, the crude mixture was purified by reverse phase HPLC (acetonitrile / water containing trifluoroacetic acid) to give the desired product (486 mg, 75% yield) as a TFA salt.

Calcd for C19H21N7O5CI2.1,5 CF ₃ CO ₂ H:

C, 39.48; H, 3.39; N, 14.65.

Found: C, 39.29; H, 3.14; N, 14.72.

NMR and MS were consistent with the desired structure.

Example 25

Preparation of N - [[5 - [(4,5-dihydro-1H-imidazol-2-yl) amino] -1H-pyrazol-3-yl] carbonyl] glycyl-3- (3,5-dichloro- 2-hydroxyphenyl) -4-alanine

The compound was prepared according to Example 24 using benzylhydrazine instead of methylhydrazine in step 2. The resulting N-benzalpyrazole compound was deprotected in the last step under catalytic hydrogenation conditions (5% Pd / C, 138 kPa, room temperature, 26 hours). This reaction was carried out using acetic acid-trifluoroacetic acid (4/1) as solvent.

Calcd. 1.75 CF ₃ CO ₂ H. 1.25H ₂ O:

· · · · «« «« «« «

C, 39.56; Η, 3.32; 13, 13.88. Found: C, 36.35; H, 2.96; N, 14.28.

NMR and MS were consistent with the desired structure.

Example 26

The above compound was prepared as described in Example 9 except that the product of Example S was used in place of the product of Example R, Step 4. Step S.

Analysis calculated for C ₂₁ H ₂₁ BrBN ₆ O 6.2 CF 3 CO ₂ H. 0.25 H ₂ O

C, 33.60; H, 2.76; N, 9.40

Found: C, 34.23; H, 2.35; N, 9.72

Example 27

(2)

Whose

Cl

Cl

Cl ·· · · ·· ·

• · · · · · · · · · · · · · · · · · · ·

The starting material necessary in this preparation was prepared according to the procedure of Example 21,

Step 1.

118 * ····

Step 1

A solution of the starting material (1.4 g) in ethanol (50 mL) was treated with a catalytic amount of 5% Pt / C at a pressure of 50 psi for 5 hours at room temperature. The reaction mixture was filtered, concentrated, and dried to give a white solid (1.1 g). ¹ H-NMR was consistent with the desired structure.

Step 2

Compound 2 was prepared according to the experimental procedure in Example 21, step 4. ¹ H-NMR was consistent with the desired structure.

Step 3

The ester group in compound 2 was hydrolyzed according to the experimental procedure in Example 21, step 5. ¹ H-NMR was consistent with the desired structure. Analysis calculated for C 22 H 24 N 6 O 7 Cl 2. 1.50 TFA. 0,25 H ₂ O:

C, 41.08; H, 3.59; N, 11.50.

Found: C, 41.03; H, 3.69; N, 11.45.

119

• 9 · 9 · · · · · · · ·

120

The starting material necessary in this preparation was prepared as described in Example 18,

Step 1.

Step 1

Compound 1 was prepared according to the procedure of Example 18,

Step 2, except that 2-methoxyethoxymethyl chloride was used as the alkylating agent instead of iodomethane. ¹ H-NMR was consistent with the desired structure.

Step 2

A solution of the product described in Step 1 (4.6 g) in methanol (50 mL) was treated with a catalytic amount of 5% Pt / C at 50 psi hydrogen for 2 hours at room temperature. The reaction mixture was filtered, concentrated and dried to give a viscous oil (4.3 g). ¹ H-NMR was consistent with the desired structure.

Step 3

A portion of the product from Step 2 (2.0 g) was guanylated using the reaction conditions described in Step 4 of Example 21 to give Product 3 as a viscous gold oil (890 mg). ¹ H-NMR was consistent with the desired structure.

Step 4

The ester group of product 3 was hydrolyzed using the experimental procedure outlined in Example 21, Step 5. ¹ H-NMR was consistent with the desired structure.

Step 5

The product of Step 4 was combined with the product of Example I followed by the procedure of Example 12, Step 4. ¹ H-NMR was consistent with the desired structure.

Step 6

The ester group of the product of Step 5 was hydrolyzed using the experimental procedure outlined in Example 21, Step 5. ¹ H-NMR was consistent with the desired structure

121 9 9 9 9 9 4 9 9 9999909 9

9 9 9 9 9 · · ·

99 99 9

Calcd for C25H3iN ₆ O9Br ₂ .1,75 TFA. 0,25 H ₂ O:

C, 37.07; H, 3.63; N, 9.10.

Found: C, 36.86; H, 3.66; N, 9.39.

Example 29

The product of Step 28, Step 5 was used as the starting material in this process.

Step 1

A portion of the starting material (425 mg) was treated with saturated anhydrous HCl in ethanol (10 mL) at room temperature for four hours. The reaction mixture was concentrated to a dry residue and the product was used in the next step without purification.

Step 2

The ester group of the crude product from Step 1 was hydrolyzed according to the procedure of Step 5 of Example 21. ¹ H-NMR was consistent with the desired structure.

Analysis calculated for C ₂₁ H 22 N ₆ O ₇ Br ₂ . 1.75 TFA. 0,25 H ₂ O:

C, 35.27; H, 2.93; N, 10.07.

Found: C, 35.21; H, 3.16; N, 10.27.

«·« * «0

The above compound was prepared according to the procedure described in Example G. In Step 2A, 3-iodo-5-bromosalicylaldehyde was used in place of 3,5-dichlorosalicylaldehyde.

Example T

The above compound was prepared according to the procedure described in Example G. In step 2A, 3-bromo-5-chlorosalicylaldehyde was used instead of 3,5-dichlorosalicylaldehyde. Similarly, in step 2B, R-phenylglycinol was used in place of S-phenylglycinol.

123

Example U

The above compound was prepared according to the procedure described in Example D. The racemic mixture obtained in Step 2 was dissolved as described below.

«• * * * * * *

124

Step 1:

To the racemic amino ester hydrochloride (50.0 g, 158.9 mmol) obtained in Step 2 of Example D in CH ₂ Cl ₂ (500 mL) and water (380 mL) was added NaHCO ₃ (38.2 g, 454). , 5 mmol). The mixture was stirred at room temperature for 10 minutes under vigorous gas evolution. A solution of benzyl chloroformate (43.4 g, 222.8 mmol) in CH ₂ Cl ₂ (435 mL) was added over 20 minutes with rapid stirring. After 40 minutes, the reaction mixture was poured into a separate funnel and the organic solution separated. The aqueous phase was washed with CH ₂ Cl ₂ (170 mL). The combined organic solution was dried (MgSO ₄ ) and concentrated in vacuo. The resulting gummy solid was suspended in hexane and collected by filtration. The tan solid was dried in vacuo to give the desired racemic product, (62.9 g, 96% yield). The single enantiomers A and B were separated by reverse phase HPLC using a chiral column. A Whelk-0 (R, R) column, 10 µm particle size and 90:10 heptane: ethanol mobile phase were used. Optical purity was determined to be > 98% using analytical HPLC with a similar solvent and similar conditions.

¹ H-NMR was consistent with the desired structure.

Step 2A

To a solution of enantiomer A, Step 1 (57.9 g, 140.4 mmol) in CH ₂ Cl ₂ (600 mL) was added via tubing trimethylsilyl iodide (33.7 g, 168.5 mmol) in CH ₂ Cl ₂ ( 125 ml). The orange solution was stirred for 1 hour at room temperature. Methanol (27.3 mL, 674.1 mmol) was added dropwise to the solution over 15 minutes. Concentration of the reaction solution in vacuo gave an orange oil. The residue was dissolved in methyl t-butyl ether (670 mL) and extracted with 1M HCl (420 mL) and water (1 χ 230 mL, 1 130 130 mL). The aqueous extracts were washed again with MTBE (130 mL). To the aqueous solution was added solid NaHCO ₃ (52.9 g, 630 mmol) in small portions. The basic aqueous mixture was extracted with MTBE (1 χ 1.2 L, 2 x 265 mL). The combined organic solution was washed with brine and 125C by concentrating in vacuo to give the desired product (28.6 g, 73% yield). ¹ H-NMR was consistent with the desired structure.

Step 2B

To a solution of enantiomer B, from Step 1, compounded from Example 1 (38.5 g, 93.4 mmol) in CH 2 Cl 2 (380 mL), was added via tubing trimethylsilyl iodide (25.0 g, 125.0 mmol) in CH ₂ Cl ₂ (80 mL). The orange solution was stirred at room temperature for 1.5 hours. Methanol (20.0 mL, 500.0 mmol) was then added dropwise and the solution was stirred for 20 minutes. Concentration of the reaction solution in vacuo gave an orange oil. The residue was dissolved in diethyl ether (450 mL) and extracted with 1M HCl (320 mL) and water (1 x 200 mL, 1 x 100 mL). The aqueous extracts were washed again with diethyl ether (100 mL). Solid NaHCO ₃ (40.1 g, 478 mmol) was added in small portions to the aqueous solution. The basic aqueous mixture was extracted with diethyl ether (1 x 1.0 L, 2 x 200 mL). The combined organic solution was washed with brine and concentrated in vacuo to give the desired product (20.8 g, 80% yield).

Analysis calculated for CHH13Cl2NO3:

C, 47.50; H, 4.71; N, 5.04.

Found: C, 47.11; H, 4.66; N, 4.93.

The product of step 2B was converted to the desired intermediate using the reagents and conditions described in steps 4 and 5 of Example G.

Example V

The above compound was prepared according to the procedure described in Example G In step 2A, 5-bromo-3-iodobenzaldehyde was used in place of 3,5-dichlorosalicylaldehyde.

Example 30

To a stirred solution of methyl 4-methoxy-2-aminopicolinate (602 mg, 3.3 mmol) (JACS, 78, 4130, 1956) in CH ₂ Cl ₂ (20 mL) was added benzoyl isothiocyanate (1 mL, Aldrich) and the reaction mixture was stirred at room temperature for 90 minutes. The mixture was diluted with ether

4

127

4 4

4444 40 (20 mL). The mixture was cooled in an ice bath, the solid was filtered, washed with ether and dried to give the title compound (828 mg, 73.27% yield) as a white solid: ¹ H NMR: 300MHz was consistent with the proposed structure.

Step 2 Preparation

och ₃

To a stirred solution of the product of Step 1 (750 mg, 2.17 mmol) in methanol (30 mL) was added NaOMe (538 mg) under nitrogen. The reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched with glacial acetic acid (0.575 mL) and stirred at room temperature for 15 minutes. The solid was filtered under vacuum, washed with ice-cold methanol and dried to give the title compound (385 mg, 73% yield) as a white solid: ¹ H NMR: 300MHz was consistent with the proposed structure.

Step 3

Preparation • ·

To a stirred solution of the product of Step 2 (364 mg, 1.5 mmol) in methanol (20 mL) was added iodomethane (0.25 mL). The reaction mixture was heated at reflux for 2 hours. The mixture was cooled to room temperature and concentrated to give the title compound (560 mg) as a white solid: ¹ H NMR: 300MHz was consistent with the proposed structure.

Step 4

Preparation

To a stirred solution of the product of Step 3 (500 mg, 1.30 mmol) in N, N-dimethylacetamide (10 mL) was added 1,3-diamino-2-hydroxypropane (123 mg, 1.37 mmol). The reaction mixture was heated at 90 ° C for 2 hours. Concentration of the mixture in vacuo afforded the crude product, which was purified by reverse phase HPLC to afford the title compound (228 mg) as a white solid. ¹ H NMR: 300MHz was consistent with the proposed structure.

129

4444 44

• 4 4 4

4 4 · • · · ΐ

4 4 4 · 44444 4 4

4 4 ·

4

Step 5 Preparation

To a stirred solution of the product of Step 4 (200 mg) in methanol (3 mL) and THF (3 mL) was added a solution of 1M NaOH (3 mL). The reaction mixture was stirred at room temperature for 1 hour. Concentration of the mixture in vacuo afforded a gummy oil which was mixed with water (2 mL) and neutralized with 1M HCl (3 mL). The mixture was concentrated in vacuo to give the crude product, which was purified by reverse phase HPLC to give the title compound (228 mg) as a white solid.

¹ H NMR: 300MHz was consistent with the proposed structure.

Step 6. Preparation

To a stirred solution of the product of Step 5 (151.25 mg, 0.5 mmol) in N, N-dimethylacetamide (5 mL) was added 4-methylmorpholine (202 mg, 2 mmol) and 1-hydroxybenzotriazole (67 mg, 0). , 5 mmol) and amino ester product from Example R (208.5 mg, 0.5 mmol). The mixture was stirred at room temperature for 5 minutes. The reaction mixture was then treated with 1- [3- (dimethylamino) propyl-3-ethylcarbodiimide (96 mg, 0.5 mmol) and 4 (dimethylamine) pyridine (10 mg). The reaction mixture was stirred for 72 hours at room temperature »»

·

130 • 9 9 9 ·

Temperature in a nitrogen atmosphere. The reaction mixture was quenched with water (1 mL) and concentrated in vacuo to give the crude product, which was purified by reverse phase HPLC to give the title compound (42 mg) as a white solid. ¹ H NMR: 400MHz was consistent with the proposed structure.

Step 7

Preparation

To a stirred solution of Step 6 product (35 mg) in methanol (3 mL) and THF (3 mL) was added a solution of 1M NaOH (3 mL). The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated in vacuo to give a gummy oil which was mixed with water (2 mL) and neutralized with 1M HCl (3 mL). The mixture was concentrated in vacuo to give the crude product, which was purified by reverse phase HPLC to give the title compound (22 mg) as a white solid. ¹ H NMR: 400MHz was consistent with the proposed structure.

Analysis calculated for C 22 H 24 N ₆ O ₇ BrCl. 2TFA, 0.5H ₂ 0:

C, 37.22; H, 3.25; N, 10.04.

Found: C, 36.91; H, 3.17; N, 10.02.

• ·

131 • fcfcfcfcfc · · * · «· · · fcfcfc · ··· ···· · · · fcfcfc · ···· · ·

Example 31

H i

.N,

HIM

Step 1 Preparation

To a stirred solution of the product of Step 4 of Example 30 (900 mg) in glacial acetic acid (10 mL) was added 48% HBr (10 mL). The reaction mixture was heated at reflux for 3 hours. The mixture was cooled to room temperature and stirred at room temperature for 18 hours. The mixture was concentrated in vacuo to give the crude product, which was purified by reverse phase HPLC to give the title compound (720 mg) as a gummy oil.

¹ H NMR: 300MHz was consistent with the proposed structure.

Step 2 Preparation

• 44 «44

132

To a stirred solution of the product of Step 1 (720 mg, 1.7 mmol) in N, N-dimethylacetamide (10 mL) was added 4-methylmorpholine (520 mg, 5.2 mmol) and 1-hydroxybenzotriazole (221 mg, 1 mmol). , 71 mmol) and amino ester product from Example R (710 mg, 1.71 mmol). The mixture was stirred at room temperature for 5 minutes. The reaction mixture was treated with 1- [3- (dimethylamine) propyl-3-ethylcarbodiimide (211 mg, 1.7 mmol) and 4- (dimethylamine) pyridine (10 mg). The reaction mixture was stirred at room temperature under nitrogen for 72 hours. Water (1 mL) was then added to the reaction mixture and concentrated in vacuo to give the crude product, which was purified by reverse phase HPLC to give the title compound (182 mg) as a white solid. ¹ H NMR: 300MHz was consistent with the proposed structure.

Step 3

Preparation

To a stirred solution of the product of Step 2 (100 mg) in methanol (3 mL) and THF (3 mL) was added 1M NaOH solution (3 mL). The reaction mixture was stirred at room temperature for 1 hour

133 • 000 • · Φ 0 • 0 ·· 00 ·· 0 • · ·

0 0 0

0 0 00 * 0 0 «0 0

0 temperature. Concentration of the mixture in vacuo afforded a gummy consistency oil to which water (2 mL) was added and neutralized with 1M HCl (3 mL). Concentration in vacuo afforded the crude product, which was purified by reverse phase HPLC to give the title compound (42 mg) as a white solid.

¹ H NMR: 300MHz was consistent with the proposed structure.

Analysis calculated for _C 21 H ₂₁ N ₆ O ₇ BrCl. 1,5TFA, 0.5H ₂ 0:

C, 37.64; H, 3.22; N, 10.97.

Found: C, 37.56; H, 3.05; N, 10.99.

Example 32

TFA

Br «

φφφφ · · φ β ·

Κ a solution of the product of Step 3, Example 9 (860 mg, 2.45 mmol) and the product of Example T (1.0 g, 2.41 mmol) in dimethylacetamide (24 mL) was added HOBT (358) at 0 ° C mg, 2.64 mmol) followed by N-methylmorpholine (0.6 mL, 5.15 mmol). The reaction solution was stirred for 15 minutes and then EDC (470 mg, 2.45 mmol) was added. The mixture was warmed to room temperature overnight. It was then concentrated in vacuo and the residue was partitioned between ethyl acetate and water. The aqueous solution was extracted with ethyl acetate and the combined organic layers were dried (Na ₂ SO ₄ ) and concentrated. The residue was purified by RP HPLC (with a starting gradient of 90:10 water / TFA: MeCN, retention time 22.5 min) to give the desired combined product comprising recovered compound B (480 mg). To this compound (Compound B) was added 1M aqueous NaOH (6 mL) and the mixture was stirred for 3 hours. The solution was acidified with TFA to pH 4. Purification by RT HPLC (starting with 95: 5 water / TFA: MeCN gradient, retention time 24.5 min) gave the desired product (160 mg, 8% yield for two steps).

Analysis calculated for C 21 H 22 BrCl N ₅ O ₆ + 2.4 TFA:

C, 36.74; H, 2.92; N, 9.96.

Found: C, 36.83; H, 3.07; N, 9.88.

¹ H NMR was consistent with the proposed structure.

135 9 9 9

A A A A A A A A

Example 33 o ₂ n,

HIM'

CO 3 CH 3 5% Pd / C ^nh 2 r

^ '^ Ν'

CO 3 CH 3

Step 1

A solution of the compound from Step 2 of Example 18 (5.72 g) in THF (80 mL) was treated with a catalytic amount of 5% Pd / C at 50 psi hydrogen pressure for 2 hours at room temperature. The reaction mixture was filtered and the residue was washed with DMF (250 mL). The filtrate was concentrated, the residue was washed with ether, filtered and air dried to give a brown powder (4.9 g), which was used directly in the next step without further purification. ¹ H NMR was consistent with the proposed structure.

Step 2

Step 1 (500 mg) was guanylated using the tris BOC reagent and conditions described in Step 4 of Example 21 to give the desired product (128 mg) as a gold glass.

¹ H NMR was consistent with the proposed structure.

· · · · · · · · · · · · · · · · · · · · · 4 · · · · · · · · · · · ·

136

Step 3

The ester group of the product of Step 2 was hydrolyzed using the experimental procedure outlined in Step 5 of Example 21. ¹ H NMR was consistent with the proposed structure.

Analysis calculated for C ₁₁ H ₁₄ N ₄ O ₄ . 1,5 TFA:

C, 38.45; H, 3.57; N, 12.81

Found: C, 38.32; H, 3.77; N, 12.80.

Step 4

The title compound was prepared using the same procedure described in

Step 4 of Example 12 by combining the product described in Step 3 with the product of Example R. ¹ H NMR was consistent with the proposed structure.

Step 5

The ester group of the product in Step 4 was hydrolyzed according to the procedure described in Step 5 of Example 12. ¹ H NMR was consistent with the proposed structure. Analysis calculated for C 21 H 22 N ₆ O ₇ BrCl. 1,5TFA. 0,25 H ₂ O:

C, 37.86; H, 3.18; N, 11.04; Cl, 4.66; Br, 10.50.

Found: C, 37.60; H, 3.26; N, 11.20; Cl, 4.79; Br, 10.19

Example 34

137

The above compound was prepared according to the procedure described in Example 33.

In step 4, the product of Example H was used in place of the product of Example R.

Analysis calculated for C 21 H 22 N ₆ O ₇ BrCl. 2.0 TFA. 0.5H ₂ O:

C, 36.49; H, 3.06; N, 10.21

Found: C, 36.10; H, 2.83; N, 10.29.

Example 35

4444444 · • 3 · ··· · *

The above compound was prepared according to the procedure described in Example 33. In Step 4, the product of Example V was used instead of the product of Example R. Analysis calculated for C ₂₁ H ₂₂ N ₆ O ₆ Br ₂ . 1.75 TFA:

C, 35.76; H, 3.03; N, 10.21 Found: C, 35.58; H, 3.07; N, 10.61.

······ iM:: * ·: · *:::

• · · · · · · · · · · · · · · ·

Example 36 p

² ) hydrolysis

2) NaOH

• • this this · this this ···

139 · · «« * · ···· · · · · · · · · · · ·

Step 1

The product from Step 1 of Example 33 (261 mg) was guanylated using the bis-boc reagent G ₂ and the reaction conditions described in Step 5 of Example 18. The product was obtained as a white solid (207 mg). The compound was then saponified using the experimental conditions described in Step 5, Example 21. 1 H NMR was consistent with the proposed structure.

Step 2

The title compound was prepared using the same procedure described in Step 4 of Example 12. The product described in Step 1 was combined with the product of Example I. ¹ H NMR was consistent with the proposed structure.

Step 3

The ester group from the product in Step 2 was hydrolyzed using the same conditions described in Step 5 of Example 12. The product described in Step 1 was coupled to the product of Example 1. ¹ H NMR was consistent with the proposed structure.

Analysis calculated for C _{2 O} H ₂ o Ν ₆ Ο ₆ Βγ ₂ .1,75 TFA. 0,25 H ₂ O:

C, 35.10; H, 2.79; N, 10.45 Found: C, 34.85; H, 2.59; N, 10.62.

140 • ΦΦΦ φφ φ φ ·

Example 37

AfNO)

CH3I SKPdZC / S k ₂ coj ** TI

COjM ·

NO. 1

NO. 1 ···· ·· ·· · ·· ··· ··· · " ·

141

Step 1

2-Nitrothiophene-4-carboxaldehyde (5.0 g) was suspended in water (100 ml) at room temperature and a solution of sodium hydroxide (5.2 g) in water (50 ml) was added in one portion. The black colored reaction mixture was stirred at room temperature for 1 hour and then filtered through a celite pad. The filtrate was acidified with 1M HCl solution and then extracted with ethyl acetate. The combined organic extracts were washed with saturated sodium hydroxide solution, dried (Na ₂ SO 4) and concentrated to give a pale yellow solid (5.0 g) which was used without further purification. ¹ H NMR was consistent with the proposed structure.

Step 2

The product of Step 1 (2.5 g) was stirred with potassium carbonate (2.0 g) and methyl iodide (2.2 g) in DMF (30 mL) for 16 hours at room temperature. The reaction mixture was partitioned between ethyl acetate and water and the layers were separated. The aqueous layer was extracted with an additional amount of ethyl acetate, and then the combined organic extracts were washed with water, saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated. The dark colored residue was purified on a silica gel column eluting with 15% ethyl acetate - 85% hexane to give a yellow solid (950 mg). ¹ H NMR was consistent with the proposed structure.

Step 3

The product obtained in Step 2 (2.0 g) was dissolved in methanol (50 mL) and treated with a catalytic amount of 5% Pd / C at 344.7 kPa under a hydrogen atmosphere at 60 ° C for 32 hours. The reaction mixture was cooled, filtered and the filtrate was concentrated. The residue was purified on a silica gel column eluting with 25% ethyl acetate - 75% hexane to give a yellow solid (920 mg). ¹ H NMR was consistent with the proposed structure.

Step 4

A solution of the product of Step 3 (900 mg) was dissolved in ethyl acetate (20 mL) and mixed with benzoyl isothiocyanate (947 mg) added at once at room temperature. Re ·················

142. · · · ί; The mixture was stirred for 30 minutes, the precipitate was filtered, washed with ethyl acetate and air dried to give a pale yellow solid (1.51 g). ¹ H NMR was consistent with the proposed structure.

Step 5

The product obtained in Step 4 of Example 35 was dissolved in methanol (70 mL) and treated at room temperature with sodium methoxide (1.3 g) added portionwise. The reaction mixture was stirred for 30 minutes and then quenched with glacial acetic acid (1.4 g). The reaction mixture was concentrated and the residue was partitioned between ethyl acetate and water. The aqueous layer was extracted with an additional amount of ethyl acetate and then the combined organic extracts were washed with brine, dried (Na ₂ SO ₄ ) and concentrated to give a pale yellow powder (900 mg).

¹ H-NMR was consistent with the proposed structure.

Step 6

The product of Step 5 (900 mg) was dissolved in methanol (20 mL) and treated with methyl iodide (1.4 g). The reaction was refluxed for 2 hours, then cooled and concentrated. The residue was mixed with ether, filtered and air dried to give a white solid (1.27 g). ¹ H NMR was consistent with the proposed structure.

Step 7

The product of Step 6 (1.2 g) was heated at 95 ° C to 100 ° C for 8 hours with 1,3-diamine-2-hydroxypropane (469 mg) and DMA (20 mL). The reaction was cooled and the solvent was removed under high vacuum. The residue was purified by reverse phase HPLC eluting with a water (0.5% TFA) -acetonitrile gradient to give the desired product (255 mg). ¹ H NMR was consistent with the proposed structure.

Step 8

The title compound was prepared using the compound produced in Step 7 under the conditions described in Step 5, Example 21. ¹ H NMR was consistent with the proposed structure.

99 · 9 9 · 9 9 9 99

-.- 999 9999 99 9

143 ·· ··· 9 ······ · 9

9999 99 9 9 9 9

Step 9

The title compound was prepared by the same procedure as described in

Step 4 of Example 12 by combining the product of Step 8 with the compound of Example G. ¹ H NMR was consistent with the proposed structure.

Step 10

The ester group of the product of Step 9 was hydrolyzed according to the procedure described in U.S. Pat

Step 5 of Example 12. ¹ H NMR was consistent with the proposed structure.

Analysis calculated for C ₂ oH ₂ and N ₅ O ₆ S ₂ C. 1.75 TFA.

C, 38.43; H, 3.19; N, 9.54; S, 4.37.

Found: C, 38.43; H, 3.39; N, 9.73; S, 4.27.

144

145

000 00 00 0

0 · · 0 0

0 000 00000 • 0 0 0 0 0

00 00 0

0

Step 1

Acetic anhydride (20 mL) was mixed with 70% nitric acid (4 mL) at room temperature. This solution was then added dropwise to a stirred mixture of N-methyl pyrrole-2-carboxylic acid (5.0 g) in acetic anhydride (30 mL) at -30 ° C. The reaction was stirred and then warmed to room temperature for 30 minutes. The reaction was then recooled to -25 ° C, then filtered through a cold filter, washed with cold hexane and air dried to give a yellow powder (1.5 g). ¹ H NMR was consistent with the desired structure.

Step 2

The title compound was prepared according to the procedure described in Step 2, Example 37 using the product prepared in Step 1. The crude product was purified by column chromatography on silica gel eluting with 40% ethyl acetate-hexane. ¹ H NMR was consistent with the proposed structure.

Step 3

The product of Step 2 (1.0 g) was dissolved in methanol (25 mL) and treated with a catalytic amount of 5% Pd / C at 50 psi under a hydrogen atmosphere for 3 hours. The reaction mixture was filtered and concentrated to give a dark red liquid (1.0 g) which was used without further purification. ¹ H NMR was consistent with the proposed structure.

Step 4

The title compound was prepared using the product described in Step 3 of Step 4 to 8 as described in Example 37. ¹ H NMR was consistent with the proposed structure.

Step 5

A solution of the product of Step 4 (425 mg), 2-chloro-4,6-dimethoxytriazine (201 mg) and N-methylmorpholine (263 mg) in DMA (10 mL) was stirred in an ice bath under nitrogen atmosphere. The reaction was stirred and warmed to room temperature for 3 hours. A solution of the product of Example G (386 mg) was prepared and N-methylmorpholine (105 mg) in DMA (5 ml) was added to the reaction mixture at room temperature at once. The reaction was stirred for 15 hours, quenched with TFA (1.5 mL) and then concentrated under high vacuum. The residue was purified by

4445 4 «• 4 4

I46. :::

4 «« 4 • 44 • 44 • 4 4 4 4 4 4

4 4 4 4 4

4 «4444444 4 · • * 4 4 4 *« 4 4 «4,444

Reversed phase HPLC eluting with water (0.5% TFA) -acetonitrile to give a white solid (706 mg). ¹ H-NMR was consistent with the proposed structure.

Step 6

The ester group of the product of Step 5 was hydrolyzed according to the procedure described in Step 5 of Example 12. ¹ H NMR was consistent with the proposed structure.

Calcd for C ₂₁ H ₂₄ N ₆ O ₆ Cl ₂ 1.5 TFA. 0.5 H ₂ O

C, 40.75; H, 3.78; N, 11.88; Cl, 10.02.

Found: C, 40.40; H, 3.68; N, 12.10; Cl, 10.20.

• o 9

147

9999 8 «• 4 9 • 9»

9 · <9 9 9 • 9 99

9 • • 9 9 9 999 99999 • 9 9 • 9 9

9

9

9

9

Example 39

Cl

C1 <<· »» »» »» »» »» » · · ·

148

O

Step 1

A mixture of 3-bromo-4-methylthiophene, (10 g) copper (I) cyanide (11.3 g) and HMPA (15 mL) was heated at 130 ° C to 140 ° C for 18 hours. The mixture was cooled, poured into a stirred solution of sodium cyanide (18.8 g) in water (28 mL) and stirred for 1 hour. The thick brown mixture was dissolved in additional water and extracted with ether. The combined ether extracts were washed with water, saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated. The residue was purified on a silica gel column eluting with 90% hexane -10% ethyl acetate to give a pale yellow liquid (5.2 g).

¹ H-NMR was consistent with the proposed structure.

Step 2

The nitrile mixture prepared in Step 1 (1.5 g) with 2M sodium hydroxide solution (150 ml) was refluxed for 1 hour. The reaction was cooled and acidified to pH 2 with 2M hydrochloric acid and extracted with ether. The combined ether extracts were dried (Na ₂ SO ₄ ) and concentrated to give a white solid (16.4 g) which was used without further purification.

¹ H-NMR was consistent with the proposed structure.

Step 3

The product of Step 2 (16.4 g) was esterified as described in Step 2 of Example 37 and purified on a silica gel column eluting with 90% hexane -10% ethyl acetate to give the desired product (13.6 g).

¹ H-NMR was consistent with the proposed structure.

Step 4

A mixture of N-bromosuccinimide (6.3 g) and dibenzoyl peroxide (100 mg) in carbon tetrachloride (25 ml) was added over 90 minutes to a refluxing solution of the product of Step 3 (5.0 g) and dibenzoyl peroxide (100 mg) in CCI. ₄ (25 mL). After 2 hours at reflux, the reaction mixture was cooled, filtered and concentrated and purification of the residue on a silica gel column eluting with 90% hexane - 10% ethyl acetate gave the title compound (4.3 g). ¹ H-NMR was consistent with the proposed structure.

Step 5

A mixture of the product described in Step 4 (4.2 g), sodium azide (2.9 g) and DMF (50 mL) was heated at 55 ° C under nitrogen for 5 hours. The reaction mixture was cooled and partitioned between water and ethyl acetate. The layers were separated and the aqueous portion was extracted with an additional amount of ethyl acetate. The combined organic extracts were washed with saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated to give a gold oil (3.5 g) which was used without further purification. ¹ H-NMR was consistent with the proposed structure.

Step 6

A solution of the product described in Step 5 (1.0 g), 1 M sodium hydroxide solution (15 mL) and methanol (15 mL) was stirred at room temperature for 16 hours. The reaction was then treated with glacial acetic acid (2 mL) and concentrated. The residue was partitioned between water and ethyl acetate. The layers were separated and the aqueous component was extracted with an additional amount of ethyl acetate. The combined organic extracts were washed with water, saturated sodium chloride solution, dried (Na ₂ SO ₄ ) and concentrated. Drying the residue at 2 ° C for 2 hours under high vacuum gave a white solid (860 mg).

¹ H-NMR was consistent with the proposed structure.

150

• · • • • · • • 4 · • • « • · • · • • • • • • · • • · · · • · • • • • E • « • • • 4 · • · 1 • ·

Step 7

A solution of the product described in Step 6 (850 mg) in dichloromethane (10 mL) was cooled in an ice bath and mixed with a solution of 2M oxalyl chloride (5 mL) in methylene chloride added at once, followed by 2 drops of DMF. The reaction was stirred for 2 hours and then warmed to room temperature. The solution was concentrated and dried under high vacuum at room temperature for 16 hours to give a tan solid (785 mg). ¹ H NMR was consistent with the proposed structure.

Step 8

A flame-dried flask was charged with a solution of diisopropylethylamine (1.3 mL), the product of Example G (1.4 g) and DMA (10 mL) under a nitrogen atmosphere. The solution was cooled in an ice bath, and a solution of hydrochloric acid from step 7 (770 mg) in DMA (5 mL) was added dropwise. The reaction was stirred for 30 minutes until the addition was complete, and then partitioned between ethyl acetate and water. The layers were separated and the aqueous portion was extracted with an additional amount of ethyl acetate. Purification of the combined organic extracts on a silica gel column eluting with 50% ethyl acetate - 50% hexane gave a white solid (1.0 g). ¹ H-NMR was consistent with the proposed structure.

Step 9

A solution of the product of Step 8 (806 mg) in ethanol (20 mL) was heated with a catalytic amount of 5% Pt / C at 13.8 kPa pressure, a hydrogen atmosphere and room temperature for 21 hours. The reaction was filtered and concentrated and the residue was purified by reverse phase HPLC and elution in a water (0.5% TFA) -acetonitrile gradient to give a white solid (480 mg). 1 H-NMR was consistent with the proposed structure.

Step 10

The product described in Step 9 (550 mg) was guanylated and purified under the conditions described in Step 4 of Example 21 to give a white solid (206 mg). ¹ H-NMR was consistent with the proposed structure.

• · * ·: ϋ

151 · · · «· · · · · · · · · ·

Step 11

The ester group of the product in Step 10 was hydrolyzed according to the same procedure described in Step 5 of Example 12. ¹ H-NMR was consistent with the proposed structure.

Calcd for C21H23N5O5Cl2S. 1.5 TFA. 0.5 H ₂ O

C, 39.79; H, 3.55; N, 9.67; Cl, 9.79; S, 4.43

Found: C, 39.91; H, 3.58; N, 9.94; Cl, 10.06; S, 4.52

Example 40

Step 1

The product prepared in Step 3 of Example 33 (1.0 g) was combined with glycine ethyl ester hydrochloride according to the procedure described in Step 5 of Example 38 and a similar purification gave a yellow solid (1.1 g).

¹ H-NMR was consistent with the proposed structure.

152 · 9 ♦ 99 <99 · 9 * 9 9 9 <<9 9 9 99 99 99 99 99 99 99 99 99 99 ··

Step 2

The product described in Step 1 (1.0 g) was hydrolyzed according to the procedure described in Step 5 of Example 21 to give a white solid (910 mg).

¹ H-NMR was consistent with the proposed structure.

Step 3

The product of Step 2 was combined with 3-amino-4,4,4-trifluorobutyric acid hydrochloride ethyl ester according to the procedure described in Step 5 of Example 38. ¹ H-NMR was consistent with the proposed structure.

Step 4

The ester group of the product in Step 3 was hydrolyzed according to the same procedure described in Step 5 of Example 12. ¹ H-NMR was consistent with the proposed structure.

Analysis calculated for C ₁₇ H ₂₀ N ₅ O ₆ F ₃ .1,5 TFA C, 38.01; H, 3.67; N, 11.08

Found: C, 37.88; H, 3.64; N, 11.02

Step 1

The product of Step 3 of Example 33 (527 mg, 1.3 mmol) and CDMT (244 mg, 1.4 mmol) was dissolved in DMAC (8 mL) under an argon atmosphere. The solution was cooled to 0 ° C for 3 hours and then NMM (0.15 mL, 1.3 mm) was added dropwise. The solution was stirred at 0 ° C for 3 hours and then the product of Example U and NMM (0.15 mL) in DMAC (10 mL) were added. The reaction was warmed to room temperature and stirred overnight at room temperature. The reaction was quenched with TFA (2 mL) and concentrated in vacuo. The crude yellow solid product was purified by RT-HPLC eluting with a 90:10 H ₂ O / TFA: CH ₃ CN gradient (λ = 254 nm). The product was isolated as a white solid (725 mg, 75% yield).

Calcd for C 23 H 26 N ₆ O ₇ Cl ₂ .1.5 TFA C, 42.12; H, 3.74; N, 11.35

Found: C, 42.26; H, 3.87; N, 11.45

HRMS M + 1 calcd for C ₂₃ H ₂₇ N ₆ O ₇ Cl ₂ 569.1318. Actual value 569.1323.

Step 2

The product of Step 1 (725 mg, 0.98 mmol) was dissolved in THF (5 mL) / 1CH ₃ OH) and 1M NaOH (6.5 mL) was added. The reaction was stirred at room temperature overnight and then 1M HCl (6.5 mL) was added. After concentration in vacuo, the crude yellow foam was purified by RT-HPLC eluting with a 95: 5 H ₂ O / TFA: CH ₃ CN gradient (λ = 254 nm). The product was isolated as a white solid (589 mg, 79% yield).

Analysis calculated for C 21 H 22 N ₆ O ₇ Cl ₂ . 1.9 TFA

C, 39.30; H, 3.18; N, 11.09 Found: C, 39.22; H, 3.16; N, 11.39

HRMS M + 1 calcd for C ₂₁ H ₂₃ N ₆ O ₇ Cl ₂ 541.1005. Actual value 541.1000.

Example 42

O

Cl • 4

154

Step 1

2-Methoxy-6-chloropyridinecarboxylic acid (10 g; 0.053 mol) was dissolved in an excess of concentrated aqueous ammonium hydroxide in a high pressure reactor and a pressure of 3447.5 kPa was reached by heating at 175 ° C for 24 hours. The solution was evaporated to dryness under reduced pressure and converted to the acid by treatment with 2M HCl at 25 ° C for 2 days and then evaporated to dryness under high vacuum at 60 ° C. After 12 hours the remaining water was azeotroped with absolute ethanol, the residue was mixed with methanol and evaporated three times under reduced pressure. The resulting residue was dissolved in methanol (250 mL), 4M HCl in dioxane (25 mol) was added to the solution, and then refluxed for 2.5 days. The solvent was removed by evaporation under reduced pressure and the crude residue was purified by reverse phase HPLC (gradient, 95/5 0.1% TFA in H ₂ O / CH ₃ CN-60/40 0.1% TFA in H ₂ O (CH ₃ CN). The desired product was dissolved in methanol, mixed with solid sodium carbonate, filtered and the solvent was evaporated. The resulting solid was suspended in ethyl acetate / hexane 50/50 and the solid collected by filtration to give the desired brown solid (1.7 g, 20% yield).

Step 2

The compound from Step 1 (300 mg; 1.8 mmol) was dissolved in DMF (7.1 mL), and the product from Step 1 of Example 9 (927 mg; 2.1 mmol), triethylamine (0.44 mL) was added. , 325 mg; 3.2 mmol) and mercuric chloride (326 mg; 1.2 mmol). The reaction mixture was stirred at 25 ° C for 3.5 hours and then heated at 60 ° C for 16 hours. After cooling to room temperature, ethyl acetate (7 mL) was added to the reaction and the heterogeneous solution was filtered through celite. Celite was washed thoroughly with ethyl acetate / ethanol 97/3. The solvents were removed under reduced pressure and the resulting red colored oil was crude dissolved in dichloromethane (30 mL), mixed with trifluoroacetic acid (2.77 mL) and refluxed for 1 hour. The reaction was cooled and removal of the solvent under reduced pressure gave a brown oil. Purification of the reverse phase HPLC sample (C18, gradient 90/10 0.1% TFA / H ₂ O) gave the impure compound, which was dissolved in methanol (2 mL), mixed with 1M NaOH (0.15 mL), and drying to dryness gave a dark brown oil. This oil was purified by reverse phase HPLC (C18, gradient 99/1 0.1% TFA / H ₂ O / CH ₃ CN) to give a tan solid (36 mg).

0 0 · · ·

155 • · · ·

0 0 0 0 0 0

000 · 000 0 0 0 0 0 0 0 00 00 00 0

Step 3

The compound from the previous step (74 mg; 0.18 mmol) was dried overnight under vacuum and dissolved in DMF (1ml) (stored over molecular sieve) and cooled to 0 ° C. Then N-methylpiperidine (27 mL; 22 mg; 0.22 mmol) was added followed by isobutyl chloroformate (47 mL; 49 mg; 0.36 mmol) and the mixture was stirred for 5 minutes. A solution of the product of Example G (0.2 mL) in DMF was added and the residual β-aminoester was washed with additional DMF (0.2 mL). Additional N-methylpiperidine (22 mL; 18 mg; 0.18 mmol) was added. The reaction mixture was slowly warmed to room temperature. After stirring at 25 ° C for 12 hours, the solvent was removed under high vacuum and a brown-red oil was obtained, which was purified by reverse phase HPLC (C18, gradient 80/20 0.1% TFA / H ₂ O / CH ₃ CN), obtained the combined product (22 mg) and this combined isobutylformyl product was coupled to the pyridine oxygen atom (47 mg). These products were mixed and dissolved in methanol (2 mL) to which was added 1M aqueous sodium hydroxide (240 mL). The reaction was stirred overnight at 25 ° C and then trifluoroacetic acid (84.7 mL) was added. The solvent was removed under reduced pressure and purified by reverse phase HPLC (C18, gradient 95/5 0.1% TFA / H ₂ O / CH ₃ CN) to give the desired product (37 mg). Analysis calculated for C ₂₁ H ₂₂ N ₆ O ₇ Cl ₂ . 2.1 TFA. 0.2 H ₂ O:

C, 38.58; H, 3.15; N, 10.71.

Found: C, 38.12; H, 3.36; N, 10.71.

Example 43

The above compound was prepared according to the procedure described in Step 3, Example 42, by reacting the product of Step 2, Example 42 with the product of Example R. Microanalysis calculated for C ₂₁ H ₂₂ NeOyCIBr. 1,8 TFA:

C, 37.35; H, 3.03; N, 10.62.

•

156 • A · A · · · · ·

9 9 · A A A A A 9 9 9 9 9 9 9 9

Found: C, 37.31; H, 3.23; N, 10.65.

Example 44

Step 1

The product from Step 3, Example 17 (3.0 g; 5.77 mmol) was dissolved in isopropanol (25 mL) and concentrated ammonium hydroxide (45 mL) was added. After stirring at 25 ° C for 3 hours, the solution was evaporated to dryness under reduced pressure. The raw material was hydrogenated in a Paar vessel in ethanol (3A) (50 mL) in the presence of 5% platinum on carbon for 2.5 hours at 25 ° C and 34.5 kPa. The sample was filtered and the filtrate was concentrated to dryness under reduced pressure. The compound was purified by reverse phase chromatography (C18, gradient 80/20, CH ₃ CN / H ₂ O) to give the desired product (1.5 g, 39% yield).

Step 2

The product of Step 1 (200 mg) was dissolved in methanol (2 mL) and sodium bicarbonate (133 mg) was added. The solution was stirred at 25 ° C for 30 min, filtered and the resulting solution was evaporated to dryness. The resulting material (205 mg; 0.43 mmol) was dissolved in DMF (1.72 mL), then the product of Step 1, Example 9 (282 mg; 0.65) was added, followed by triethylamine (99 mg; 97 mmol, 0.14 mL) followed by mercuric chloride (177 mg; 0.65 mmol). The solution was heated at 60 ° C for 30 minutes and then at 90 ° C for 12 hours. The reaction mixture was cooled, then dissolved in ethyl acetate (1.7 mL) and filtered through celite. The resulting filtrate was evaporated to dryness. The crude material was purified by flash chromatography for ODI.

157 removal of unreacted starting material. This material was reacted with 50/50 TFA / CH ₂ Cl ₂ to give the unpurified desired product. This material was dissolved in methanol (2 mL) and 1M sodium hydroxide (2 mL). After stirring at 25 ° C for 12 h, the solution was neutralized with TFA (77 mL) and then evaporated to dryness under reduced pressure. The crude product was purified by reverse phase chromatography (C18, 95/5 H ₂ O / CH ₃ CN) to give the desired product (6.8 mg).

Microanalysis calculated for C21H23N7O6Cl2. 3,3TFA:

C, 36.17; H, 2.89; N, 10.70.

Found: C, 36.57; H, 3.21; N, 10.37.

· «* Φ φ · ·

158 φ φ φ φ φ φ • • φ • • φ •

Example 45

1. guanylation

2. hydrolysis

Step 1

To a mixture of 3-methyl-2-thiophenecarboxylic acid (1.0 g, 7.0 mmol), N, N-dimethylformamide (4.0 mL) and potassium carbonate (1.93 g, 14 mmol) was added iodomethane (1 mL). , 49 g, 10.5 mmol). The reaction was stirred at room temperature for 80 hours, diluted with ethyl acetate (150 mL) and washed with H ₂ O (100 mL) and brine (100 mL). The organic layer was dried (Mg ₂ SO ₄ ) and concentrated to give the pure product as a brownish oil (0.87 g, 81% yield). ¹ H-NMR was consistent with the desired structure.

159

999999 »99 9 · 9 · 9 9 9 9 · · · 9

999 9999 · 9 9

9 * 9 9999999

9900 99 99 99 9

99 99 99 99 999

Step 2

To a solution of the product of Step 1 (5.0 g, 32.0 mmol) dibenzoyl peroxide (0.08 g) and carbon tetrachloride (20 mL) was added a mixture of N-bromosuccinimide (6.3 g, 35.3 mmol), dibenzoyl peroxide (0.08 g) and carbon tetrachloride (20 mL) and refluxed for 30 minutes. The resulting reaction mixture was heated at reflux for 18 hours. The solid was filtered and washed with carbon tetrachloride (2 × 10 mL). Concentration of the filtrate gave an oil-solid mixture (8.1 g, 80% yield). ¹ H-NMR was consistent with the desired structure.

Step 3

A mixture of the product of Step 2 (8.1 g, 34.7 mmol), sodium azide (5.6 g, 87 mmol) and Ν, Ν-dimethylformamide (25 mL) was heated at 58 ° C for 2.5 h. The reaction was diluted with ethyl acetate (800 mL) and washed with H ₂ O (500 mL). The aqueous layer was extracted with ethyl acetate (100 mL). The obtained organic layer was dried (Mg ₂ SO ₄ ) and concentrated to give a brownish oil (6.0 g, 85% yield). ¹ H-NMR was consistent with the desired structure.

Step 4

A solution of the product of Step 3 (6.0 g, 25.9 mmol), NaOH (1M, 50 mL) and MeOH (50 mL) was stirred overnight. The reaction was mixed with acetic acid (2.5 mL). The product was extracted with ethyl acetate (300 mL). The organic layer was dried over Na ₂ SO ₄ and concentrated to give a yellow oil. A yellow solid was obtained in vacuo (2.0 g, 42% yield). ¹ H-NMR was consistent with the desired structure.

Step 5

A mixture of Step 4 product (0.5 g, 2.7 mmol) with oxalyl chloride (2M in dichloromethane, 2.7 mL, 5.4 mmol) and dichloromethane (20 mL) was stirred at room temperature for 2.5 h. . Removal of the solvent from the reaction gave the product as a brown oil (0.51 g, 85% yield). ¹ H-NMR was consistent with the desired structure.

fefe *

160 • fe 99 fefe · fe • fefe · 9 «fefe · fefe fefe fe ·· • fefe · · * fe · · · • fefe · · ·

Step 6

To a mixture of the product of Step 5 (0.46 g, 2.43 mmol) and THF (10 mL) at 0 ° C was added a solution of the product of Example G (0.91 g, 2.43 mmol), diisopropylethylamine (0). , 63 g, 4.86 mmol) and N, N-dimethylacetamide (20 mL). The reaction mixture was stirred at 0 ° C for 10 minutes and then warmed to room temperature. After 16 hours, the solvent was removed from the reaction in vacuo. The product was extracted with ethyl acetate (300 mL). The organic solution was washed with saturated NaHCO ₃ (100 mL), H ₂ O (100 mL) dried over Na ₂ SO _4, and concentrated to give the crude product. The crude product was purified by column chromatography (on silica gel, CH ₂ Cl ₂ / MeOH / NH ₄ OH, 98/2 / 0.2) to give the pure product as a gummy solid (0.7 g, 58% yield) . ¹ H-NMR was consistent with the desired structure.

Step 7

A mixture of the product of Step 6 (0.78 g, 1.56 mmol), 5% Pt / C and EtOH was stirred for 20 hours at room temperature and a pressure of 34.5 kPa. The catalyst was filtered off. The filtrate was mixed with trifluoroacetic acid (0.6 mL) and concentrated to give the crude product. Purification of the crude product by HPLC gave a white solid (0.4 g, 54% yield).

¹ H-NMR was consistent with the desired structure.

Step 8

To a mixture of the product of Step 7 (0.55 g, 1.25 mmol), the product of Step 1, Example 9 (0.81 g, 1.87 mmol), triethylamine (0.38 g, 3.75 mmol) ) and N, N-dimethylaformamide (15 mL) were added mercuric chloride (0.51 g, 1.87 mmol) at room temperature. The reaction was heated at 95 ° C to 100 ° C for 16 hours. The cooled reaction mixture was filtered through a pad of celite (2 "), and washed with ethyl acetate. Removal of the solvent from the filtrate under vacuum gave an oil. The oil was diluted with CH ₂ Cl ₂ / trifluoroacetic acid (10 mL / 10 mL) and stirred for 1 hour at room temperature. The solid formed was removed by filtration. The filtrate was concentrated and treated with CH ₂ Cl ₂ / trifluoroacetic acid (15 mL / 15 mL) for 1.5 h. Removal of the solvent under reduced pressure gave a brown oil. Purification of this oil by HPLC gave a mixture of the desired product and its ethyl ester. The title compound was recrystallized from acetonitrile as a white solid

161

445 * «4

4 4

4 ·

4 ·

4 4 ·

4 4 4

4

4 4 * 4 4 * * 4444

4 4

4 * compounds (0.068 g). The ester product was treated with NaOH / ethanol (7.0 mL / 7.0 mL) for 18 h and purified by HPLC to give additional compound (0.055 g).

Calcd for C21H23N5O6Cl2S. 2.0 CF ₃ COOH;

C, 38.87; H, 3.26; N, 9.07,

Found: C, 38.78; H, 3.6; N, 9.16.

Example 46

Ό

• · · · · · · · · · · · · · · · · ·

162

Step 1

To a solution of the product of Step 1 of Example 33 (3.5 g, 20.8 mmol) and N, N-dimethylformamide (45 mL) was added benzoyl isothiocyanate (3.34 g, 21.3 mmol) at room temperature. The reaction was stirred for 2 hours and poured into ether (300 mL). The resulting mixture was stirred for 15 minutes. The product was collected by filtration, washed with ether (2 x 40 mL) and air dried to give a yellow solid (6.95 g, 100% yield). ¹ H-NIVIR was consistent with the desired structure.

Step 2

To a suspension of the product of Step 1 (6.90 g, 20.8 mmol) and methanol (200 mL) was added sodium methoxide (2.64 g, 48.9 mmol) in small portions at room temperature. Acetic acid (2.8 mL, 48.9 mmol) was added to the reaction. The resulting mixture was stirred for 15 minutes. The product was collected by filtration, washed with methanol (2 × 30 mL) and dried under reduced pressure to give an off-white solid (3.8 g, 81% yield). ¹ H-NMR was consistent with the desired structure.

Step 3

A mixture of the product of Step 2 (1.90 g, 8.4 mmol), methyl iodide (3.56 g, 25.1 mmol) and methanol (60 mL) was heated at reflux for 3.5 hours and cooled to room temperature. Removal of the solvent from the reaction solution under reduced pressure gave a yellow oil. Combine the residual oil with ether to give the pure product as a yellow solid (3.1 g, 100% yield). ¹ H-NMR was consistent with the desired structure.

Step 4

A solution of the product of Step 3 (0.5 g, 1.36 mmol), 2-methoxyethylamine (0.12 mL, 1.36 mmol) and N, N-dimethylacetamide (5.0 mL) was heated for 2.5 hours at 85 ° C. Additional 2-methoxyethylamine (0.12 mL, 1.36 mmol) was added. The reaction was heated at 85 ° C for 2 hours. Removal of Ν, Ν-dimethylacetamide from the cooled reaction mixture under reduced pressure gave the crude product. The crude product was purified by reverse phase HPLC to give the desired purified product (0.38 g, 76% yield). ¹ H-NMR was consistent with the desired structure.

···· 44 • ·

4 • 44

4 4

444

• 4 4 4

44

4 4

4 4 4 • 4 44 ·

4 4

4

Step 5

A solution of the product of Step 1 (0.38 g, 1.42 mmol), a solution of NaOH (1M, 15 mL) and MeOH (15 mL) was stirred for 16 hours at room temperature, and then trifluoroacetic acid (1.2 mL) was added. ). Removal of the solvent under reduced pressure gave the crude product. The crude product was purified by HPLC to give the desired product as a white solid (0.55 g, 100% yield). ¹ H-NMR was consistent with the desired structure.

Step 6

To a solution of the product of Step 5 (0.54 g, 1.42 mmol) in N, N-dimethylacetamide (16 mL) was added N-methylmorpholine (0.1 mL, 1.42 mmol). To the reaction mixture was added isobutyl chloroformate (0.18 g, 1.35 mmol) at -5 ° C over 5 minutes and stirred for 15 minutes. The product of Example G (0.396 g, 1.07 mmol), N-methylmorpholine (0.075 mL, 1.07 mmol) and N, N-dimethylacetamide (10 mL) were added to the reaction. The resulting reaction solution was warmed to room temperature and stirred for 16 hours. Removal of the solvent from the reaction solution under reduced pressure gave the crude product. The crude product was purified by HPLC to give the desired product as a white solid (0.145 g, 14% yield). ¹ H-NMR was consistent with the desired structure.

Step 7

A solution of the product of Step 6 (0.145 g, 0.20 mmol), NaOH (1M, 25 mL) and MeOH (25 mL) was stirred for 16 hours. Removal of the solvent from the reaction mixture under reduced pressure gave the crude product. The crude product was mixed with trifluoroacetic acid (2 mL) and purified by HPLC to give the desired product as an off-white solid (0.11 g, 70% yield).

Analysis calculated for C 21 H 24 N 6 O 7 Cl 2 CF ₃ COOH:

C, 38.93; H, 3.40; N, 10.89;

Found: C, 39.29; H, 3.47; N, 11.15.

164 ·····································

Example 47

Step 1

A mixture of the product of Step 3 of Example 46 (1.56 g, 4.22 mmol), 2,2-dimethyl-1,3-propanediamine (0.45 g, 4.40 mmol) and N, N-dimethylacetamide (15 mL). ml) was heated to 6.5

165

ΦΦΦΦ · · φ φ φ φ «« «« φ φ φ φ φ φ φ • • Φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ pak pak pak pak hodiny hodiny při při 92 92 92 92 A solid formed overnight. The product was obtained by filtration, washing with N, N-dimethylacetamide (5 mL) and ether (5 mL) to give the pure product as a white solid (0.50 g, 43% yield).

¹ H-NMR was consistent with the desired structure.

Step 2

A solution of the product of Step 1 (0.48 g, 1.7 mmol), NaOH (1M, 10 mL) and MeOH (10 mL) was stirred overnight at room temperature. Removal of the solvent from the reaction mixture under reduced pressure gave the crude product. The crude product was mixed with trifluoroacetic acid (2 mL) and purified by HPLC to give the desired product as a white solid (0.38 g, 58% yield).

Step 3

To a solution of the product of Step 2 (0.36 g, 0.94 mmol), 2-chloro-4,6-dimethoxytriazine (0.18 g, 1.03 mmol) and N, N-dimethylacetamide (10 mL) at A solution of 4-methylmorpholine (0.14 g, 1.41 mmol) in N, N-dimethylacetamide (1.5 mL) was added at 0 ° C. The resulting pale orange solution was warmed to room temperature and stirred for 3 hours. To the reaction solution was added a solution of the product of Example G (0.350 g, 0.94 mmol), 4-methylmorpholine (0.93 g, 0.94 mmol) and N, N-dimethylacetamide (4 mL). The resulting solution was stirred overnight at room temperature and mixed with trifluoroacetic acid (1.0 mL). Removal of the solvent from the reaction mixture under reduced pressure gave the crude product. Purification of the crude by HPLC gave the desired product as a yellow solid (0.35 g, 54% yield).

Step 4

A solution of the product of Step 3 (0.33 g, 0.47 mmol), NaOH (1M, 9 mL) and MeOH (9 mL) was stirred for 18 hours. The reaction was mixed with trifluoroacetic acid (1.0 mL). Removal of the solvent from the reaction mixture under reduced pressure gave the crude product. Purification of the crude by HPLC gave the desired product as a yellow solid (0.285 g, 84% yield).

Analysis calculated for C ₂₃ H ₂₆ Ν ₆ Ο ₆₀ 0Ι ₂ . 1.25 CF ₃ COOH. 1,0H ₂ O:

C, 42.90; H, 4.13; N, 11.77;

Found: C, 43.03; H, 4.03; N, 11.26.

166

The above compound was prepared according to the procedure described in Example 47. In Step 1, ethylenediamine was used instead of 2,2-dimethyl-1,3-diaminopropanediamine. In step 3, the product of Example U was used instead of the product of Example G.

Analysis calculated for C ₂₀ H ₂₀ N ₆ O ₆ Cl ₂ . 1.25 CF ₃ COOH. 0.5H ₂ 0:

C, 40.77; H, 3.38; N, 12.68;

Found: C, 40.98; H, 3.17; N, 12.57.

♦ · 4 e 4 4 4 4 4 4 4 4 4

167 • ·· t · t · ♦

4 4 4 5 6 7 8 9

Example X

Preparation of 5-bromo-3-iodosalicylaldehyde

CHO

The above compound was prepared by iodination of 5-bromosalicylaldehyde as described in the literature (J. Org. Cherm. 1990, 55, 5287-5291).

Examples of Y & Z

Scheme A1

Benzoyl isothiocyanate acetinitrile, reflux for 16 hours

1. NaOMe, MeOH RT, 2.5 hr

2. Mel, CH 3 COOH

DMF, 90%: 3-4h

Ri

OH

Z_ £ Rt ³ Rj β CH) X- R, H, R ₂ = OH • ·

168 • · · · · ·

Step 1

Preparation

A mixture of 5-aminonicotinic acid (10.0 g, 0.072 mol), benzoyl isothiocyanate (11.8 g, 0.072 mol) and DMAP (catalytic amount) in anhydrous acetonitrile (250 mL) was refluxed overnight under anhydrous conditions and vigorous stirring (scheme) A1). The resulting yellow suspension was cooled and filtered. The residue was washed with water, then with acetonitrile and dried under vacuum overnight to give the desired product as a pale yellow solid (21.4 g, 98% yield).

MS and ¹ H-NMR were consistent with the desired structure.

Step 2 Preparation

HN

To a suspension of the product of Step 1 (11.1 g, 0.037 mol) in anhydrous MeOH (230 mL) was added NaOMe (25 wt% solution in methanol, 21.1 mL, 0.092 mol), the reactants entered the solution and formed orange-brown solution (Scheme A1). The solution was stirred at room temperature for 3 hours, cooled in an ice bath, and methyl iodide (3.45 mL, 0.055 mol) was added. The resulting mixture was stirred at 10 ° C for 30 minutes and at room temperature for 1.5 hours.

4 4

169

4 4

4 4 ·

4 4 4 4

4444444 4 temperature. The reaction mixture was then quenched with acetic acid (2 mL), cooled in an ice bath, and filtered. The solids were washed with cold MeOH and dried in vacuo to give the desired product as a beige solid (2.66 g, 37% yield).

MS and ¹ H-NMR were consistent with the desired structure.

Step 3

Preparation of Y & Z examples

To a solution of 1,3-diamine-2-hydroxypropane (11.2 g, 0.124 mol) in anhydrous DMF (80 mL) was added the product of Step 2 (8.7 g, 0.041 mol). The mixture was heated at 85 ° C for 3 hours under anhydrous conditions (Scheme A1). After 1-2 hours, the solution became cloudy and turbidity increased during heating. The reaction mixture was then cooled in an ice bath and filtered. The solids were washed with acetonitrile, water, acetonitrile and dried in vacuo to give the desired product (Example Y) as a beige solid (3.7 g, 38% yield).

MS and ¹ H-NMR were consistent with the desired structure.

Example Z was synthesized according to the procedure described in Example Y by replacing 2,2-dimethyl-1,3-propanediamine with 1,3-diamine-2-hydroxypropane.

Each of the products of Step 3 was converted to their TFA or HCl salts by stirring at 10 ° C for 1 h in a solution of anhydrous THF (10 mL per 1.0 g substrate) and TFA (1 equivalent) or 4M HCl / dioxane (2 mL). equivalents).

Example 49

Preparation

Rj * Rt ^w Me

170

isobutyl formate

NMM, DMF, -15 ° C - RT 3h 2. 1MOHOH.RT, 1h, HCl

C.

d.

X = Y = Cl R | -R ₂ ~ CH ₃ Χ-Cl, Y-Br R, = R ₂ = CH ₃

To a suspension of Example Y (0.40 g, 0.00125 mol, Scheme A2) in anhydrous DMF (10 mL) at -20 ° C was added isobutyl chloroformate (0.17 g, 0.00125 mol) followed by dropwise addition. N-methylmorpholine (0.14 g, 0.00137 mol) was added. After stirring at 20 ° C for 20 minutes, additional N-methylmorpholine (0.14 g, 0.00137 mol) was added, followed by the addition of the product of Example G (0.46 g, 0.00125 mol). The resulting mixture was stirred at 20 ° C for 15 minutes and then stirred at room temperature for 2 hours. DMF was distilled off under vacuum and purification of the residue by reverse phase HPLC gave (after lyophilization) the desired ester as a white solid (0.20 g, 21% yield).

MS and ¹ H-NMR were consistent with the desired structure. The ester (0.2 g) was stirred for 1 hour with 1M LiOH (2 mL) at room temperature. The pH was adjusted to 2 with trifluoroacetic acid and purification by reverse phase HPLC gave (after lyophilization) the desired acid as a white solid (0.11 g).

MS and ¹ H-NMR were consistent with the desired structure.

171

Example 50

Preparation

Rj = Rj ** Mc

The above compound was prepared (Scheme A2) according to the procedure described in Example 49 by replacing an equivalent amount of Example R with the product of Example G.

MS and ¹ H-NMR were consistent with the desired structure (s). The ester (0.19 g, 0.00023 mol) was stirred for 1 hour with 1M LiOH (2 mL) at room temperature. The pH was adjusted to 2 with trifluoroacetic acid and purification by reverse phase HPLC gave (after lyophilization) the desired acid as a white solid (0.13 g, 72% yield).

MS and ¹ H-NMR were consistent with the desired structure.

Example 51

Preparation

• · · · · ·

172

Scheme Β

Z-ONSu, NMM RT, 16h. NH-Z

DMSO / EDC. NH-Z

CH ₂ Cl ₂

NH-Z pyridinium trifluoroacetate

AND.

NH-Z ho ^xx '* ^<oh —-.......

pTSA

H ₂ Pd / C (5%)

EtOH, EtOAc h

dichloroethane, DMSO 80 ° C, 24 h

348.5 kPa

NH

DMF, Et ₃ N 90 ° C, 3h

Step 1

Preparation of bis-N-benzyloxycarbonyl-2-hydroxy-1,3-diaminpropane.

.NH-Z ho ^ ^nh · ² * · · · · · φ

Κ a suspension of 2-hydroxy-1,3-diaminopropane (5.8 g, 0.064 mol) in dichloromethane (150 mL) containing N-methylmorpholine (14 mL) was added portionwise at 10 ° C benzyloxycarbonylsuccinimide (32 g, 0.129 mol) (Scheme B). The reaction mixture was stirred at room temperature for 16 hours and the resulting clear solution was diluted with dichloromethane (100 mL), washed sequentially with 10% citric acid (2 x 50 mL), water, and dried (Na ₂ SO ₄ ). After filtration, the solvent was removed under reduced pressure and recrystallized from dichloromethane to give the title compound (20.0 g, 87% yield).

¹ H-NMR and MS were consistent with the desired structure.

Step 2

Preparation of bis-N-benzyloxycarbonyl-1,3-diaminpropan-2-one.

NH-Z

NH-Z

To a suspension of the product of Step 1 (21.0 g, 0.058 mol) and EDC (33.0 g) in dichloromethane (120 mL) containing DMSO (24.0 mL) was added dropwise a solution of pyridinium trifluoroacetate (33.0 g) in dichloromethane (50 mL) for 30 min and stirred at 10 ° C. A clear yellow solution was obtained after the addition. After stirring at room temperature for 3 hours, a white solid separated from the reaction mixture. The mixture was cooled, filtered and the solid washed with cold dichloromethane and water. The obtained white solid was dried in a desiccator under vacuum to give the desired product (15.5 g, 74% yield).

¹ H-NMR and MS were consistent with the desired structure.

• ·

174

Step 3 Preparation of .NH-Z \ -o

A mixture of the product of Step 2 (2.5 g, 0.007 mol), p-toluenesulfonic acid (0.3 g, 0.0016 mol) and DMSO (1.0 mL) in dichloromethane (25 mL) containing ethylene glycol (2 mL). 0 mL) was refluxed under anhydrous conditions (Scheme B). After 24 hours, the reaction mixture was cooled, diluted with dichloromethane (25 mL) and washed successively with 10% sodium carbonate, water, and dried (Na ₂ SO 4). After removal of the solvent under reduced pressure, the residue was recrystallized from dichloromethane / hexane to give the title compound (2.5 g, 89% yield).

¹ H NMR and MS were consistent with the desired structure.

Step 4

The product of Step 3 (3.0 g, 0.0075 mol) was dissolved in a solvent of a mixture of ethanol (50.0 mL) and ethyl acetate (50.0 mL) and hydrogenated at 344.75 kPa for 16 hours in the presence of Pd / C (5%, 1.5 g) at room temperature (Scheme B). The catalyst was removed by filtration, washed with 40% water / ethanol (50 mL) and filtered. The resulting filtrate and washings were concentrated to dryness under reduced pressure to give a syrupy substance. This was dissolved in DMF (10.0 mL), the product of Step 2 of Example 1 (1.0 g, 0.0047 mol) was added with triethylamine (0.7 mL) and DMAP (0.05 g). The resulting mixture was heated at 90 ° C under anhydrous conditions. After 3 hours, DMF was distilled off under vacuum, the residue was mixed with water and filtered. Washed with water, acetonitrile and dried in a desiccator under vacuum gave the product of Example 5 (0.4 g, 30%) as a powder. This material was used without further purification in Step B. ¹ H-NMR and MS were consistent with the desired structure.

• · • · I ► 4 · 4 »

175 4 4 * 4 4 44 444

Example 52

Preparation

Scheme C

NMM, DMF. -J5 ®C-RT3h 2.1M LiOH, HCl

COOH • j-OH

Example 51 (0.38 g, 0.0014 mol) was suspended in dry THF (5.0 mL), trifluoroacetic acid (0.1 mL) was added, and the mixture was stirred at 10 ° C under anhydrous conditions (Scheme C). After 30 minutes, THF was distilled off under reduced pressure and the residue was dried under vacuum for 3 hours. The resulting material was dissolved in dry DMF (4.0 mL), cooled to -15 ° C, isobutyl chloroformate (0.18 mL) was added followed by Nmethylmorpholine (0.17 mL). The solution was stirred under argon for 30 minutes. To the mixture was added an amine solution made by adding N-methylmorpholine (0.17 mL) to a solution of Example R (0.51 g) in DMF (3.0 mL) at 0 ° C. The resulting mixture was stirred at -15 ° C for 30 minutes and then at room temperature for 16 hours. The solvent was removed by vacuum distillation and

1 9 9 9 9 9 9 9 9 9 The residue was purified by reverse phase HPLC using 10 to 90%. % gradient of acetonitrile / water (40 min) at a 70 mL / min elution rate.

Appropriate fractions were combined and freeze-dried to give the desired ester (0.4 g) as a fluffy white powder. This material was stirred at room temperature with lithium hydroxide (1M, 2.0 mL). After 45 minutes, the reaction mixture was cooled, diluted with water, acidified with trifluoroacetic acid and the desired acid (0.25 g) was isolated by reverse phase HPLC using 10-90% acetonitrile / water as described above. ¹ H-NMR and MS were consistent with the desired structure.

Example 53

HIM

-NH-Z

NH-Z

OH

DAST, pyridine

...... ->

CH 3 Cl 2

-50C-RT 16 h

NH-Z

NH-Z

H ₂ Pd / C (10%)

EtOH, EtOAc, ¹⁶ 348.5 kPa

_ carobenzoxy

DMF, Et ₃ N 90 C, 3h 9 9

177

Step 1

Preparation of bis-N-benzylaoycarbonyl-2-fluoro-1,3-diaminpropane.

NH-Z

To a stirred suspension of bis-N-benzyloxycarbonyl-2-hydroxy-1,3-diaminopropane (6.0 g, 0.017 mol) in dichloromethane (50 mL) and pyridine (2.7 mL) was added dropwise at -50 ° C. solution of DAST (2.5 mL) in dichloromethane (7.5 mL, Scheme D). The reaction mixture was gradually warmed to room temperature over 16 hours under an argon atmosphere. A bright yellow solution was obtained. The solution was cooled and poured into a mixture of ice, water (100 mL) and dichloromethane (50 mL). The organic phase was washed with water (2 x 50 mL) and dried (Na ₂ SO ₄ ). After removal of the solvent, the residue was purified by silica gel flash chromatography using 30% EtOAc in hexane. Appropriate fractions were combined, concentrated to dryness and recrystallization of the product from dichloromethane / hexane gave the desired fluorinated intermediate as a fluffy white powder (2.0 g). ¹ H-NMR and MS were consistent with the desired structure.

Step 2

A solution of bis-N-benzyloxycarbonyl-2-fluoro-1,3-diaminopropane (3.3 g, 0.0092 mol) obtained from step 1 in EtOAc (30 mL) and EtOH (30 mL) was hydrogenated for 16 hours in presence of Pd / C (10%, 2.7 g) at room temperature and a pressure of 344.7 kPa (Scheme D). After filtration, the catalyst was stirred with EtOH containing 40% water (50 mL) and filtered again. Concentration of the filtrate to dryness gave a syrupy solid (0.7 g). This material was suspended in DMF (8.0 mL). The product from Step 2 of Example 1 (0.7 g, 0.0033 mol), a catalytic amount of DMAP (0.01 g) was added, and the mixture was stirred at 90 ° C under anhydrous conditions for hours. DMF was distilled off under vacuum, the residue was suspended in water (25 ml) and the pH was adjusted to 4.5 by addition of 1M HCl. The resulting mixture was cooled. The solid was collected by filtration, washed thoroughly with water, acetonitrile 9 9 9 9 ·

9,999 «99

178

999999999999999999999999999999999 and drying in a desiccator under vacuum gave the desired compound as a brown powder (0.24 g). ¹ H-NMR and MS were consistent with the desired structure.

Example 54

Preparation

2. 1M LiOH. HCl

The compound of Example 53 (0.22 g) obtained previously was suspended in dry THF (4.0 mL). Trifluoroacetic acid (0.1 mL) was added and the solution was stirred at 10 ° C for 30 min and concentrated under reduced pressure. The residue was dried in a desiccator under vacuum. The obtained material was suspended in dry DMF (5 mL). Isobutyl chloroformate (0.12 mL) was added followed by N-methylmorpholine (0.11 mL). The solution was stirred at -15 ° C under an argon atmosphere (Scheme E). After 30 minutes, a solution of the amine formed by adding N-methylmorpholine (0.095 mL) to a solution of the product of Example R (0.37 g) in DMF (3.0 mL) was formed. The resulting mixture was stirred for 30 minutes at -15 ° C and then for 16 hours at room temperature

179 • 9 · · <

44

9 9

9 9 9

99999 9 · photo. DMF was distilled off under vacuum and the residue was purified by reverse phase HPLC using 10-90% acetonitrile / water. Appropriate fractions were combined and freeze-dried to give the desired ester product as a pale yellow powder (0.35 g). ¹ H-NMR and MS were consistent with the desired structure.

The resulting product (0.3 g, Scheme E) was stirred with 1M LiOH (3.0 mL) at room temperature. After 1 hour, the solution was diluted with water (3.0 mL), cooled and acidified with trifluoroacetic acid. The resulting mixture was then purified by reverse phase HPLC using 10-90% acetonitrile / water (30 minute gradient) at a flow rate of 70 mL / min. Appropriate fractions were combined and freeze-dried to give the title compound as a white powder (0.22 g). ¹ H-NMR and MS were consistent with the desired structure.

Example 55

Preparation

Br

2. TFA

Cl

9999 98 • ·

180

Scheme F

3-4h isobutylchloroformate

NMM, DMF, -15 ° C-RT, 3h 2. 1M LiOH, HCl

Step 1

Preparation

To a solution of 1,4-diamino-2,3-dihydroxybutane dihydrochloride [2.21 g, 0.012 mol] [synthesized from dimethyl-L-tartrate as described in J. Carbohydrate Chemistry, 5, (2), 183-197, [ 1986], in water (6 mL) and anhydrous DMF (10 mL) were added sodium carbonate (1.83 g, 0.017 mol). To this mixture was added the product from Step 2 of Example 1 (1.21 g, 0.006 mol) and the mixture was heated at 85 ° C for 3 hours. After cooling in an ice bath, DMF was distilled off under vacuum and the resulting residue was suspended in water. The pH was adjusted to 5.6. Lyophilization of the solution gave the desired product (0.907 g, 59% yield).

MS was consistent with the desired structure (M + H 267).

The obtained compound was converted to the HCl salt by stirring with 4M HCl / dioxane (2 equivalents) in THF (10 mL) for 1 hour at 10 ° C.

181

0000 00 • 0 0 0 0 0 0 0 0 0 0 0 0 0

0000 00 00 00

Step 2

To a suspension of the product of Step 1 (0.11 g, 0.23 mmol) in anhydrous DMF (10 mL) at -20 ° C was added isobutyl chloroformate (0.016 g, 0.12 mmol), followed by dropwise addition of N-methylmorpholine (0.013 g, 0.13 mmol, Scheme F). The resulting mixture was stirred at -20 ° C for 20 minutes, an additional amount of N-methylmorpholine (0.013 g, 0.13 mmol) was added, followed by the addition of product R (0.048 g, 0.12 mmol). The resulting mixture was stirred at -20 ° C for 15 minutes. After stirring at room temperature for 2 hours, DMF was distilled off under vacuum and purification of the residue by reverse phase HPLC gave (after lyophilization) the desired ester as a white solid (0.03 g, 33% yield).

MS (M + H 627 M + H 629) and ¹ H-NMR were consistent with the desired structure.

¹ H NMR (400MHz, CD ₃ OD): δ 8.8 (s, 1H), δ 8.5 (s, 1H), 8.1 (s, 1H), 7.4 (s, 1H),

7.2 (s, 1H), 5.6 (m, 1H), 4.1 (m, 4H), 3.7 (m, 2H), 3.6 (m, 2H), 3.3 (m, 2H) 2.9 (m, 2H).

1.2 (m, 3H).

The resulting ester (0.03 g, 0.035 mmol) was stirred with 1M LiOH (2 mL). After stirring at room temperature for 1 hour, the pH was adjusted to 2 with trifluoroacetic acid and isolation of the reverse phase HPLC product (after lyophilization) gave the desired acid as a white solid (0.001 g, 3.5% yield).

MS (M + H 599 M + H 601) and ¹ H-NMR were consistent with the desired structure.

¹ H NMR (400MHz, CD ₃ OD): δ 8.8 (s, 1H), 8.5 (s, 1H), 8.0 (s, 1H), 7.4 (s, 1H) 7.2 (s, 1H), 5.6 (m, 1H), 4.1 (m, 2H), 3.8 (m, 2H), 3.5 (m, 2H), 3.3 (m, 2H) 2.8 (m, 2H).

Example 56

Difluoranalog was prepared according to the synthetic procedure outlined in Scheme 1 below.

182

Scheme 1

NMM, DMF, -15 ° C-RT, 3h, 2.1ML1H, RT, HCl

halogen

The carboxybenzoxy-protected ketone was converted to the corresponding difluoro intermediate using DAST diethylaminosulfur trifluoride. Subsequent removal of carbobenzoxy groups (Z) by catalytic hydrogenation and coupling of the resulting diamine with the S-methylisothiourea derivative of the 5-aminonicotinic acid derivative affords the corresponding difluoro-substituted guanidine containing the nicotinic acid precursor. This compound was reacted with an amino acid glycine beta ester followed by hydrolysis and acidification to give the desired compound.

41 4

183 • 4 · 4 4 · 4 · 4 4 4 · 4 444 · 4 · 4 · 4 · 4 44 44 4

Example 57

Compounds containing 7-membered fluoro / hydroxy substituted guanines were prepared according to the following scheme from the known starting diisopropyl-L-tartrate (J. Am.Chem. Soc. 1988, 110, 7538).

Scheme 2

2. LiOH, HCl X = Y = halogen

The monosilylated intermediate was synthesized by amidation, silylation and N-desilylation. Diborane reduction of this monosilyl derivative gave the corresponding diamine dihydrochloride. Subsequent condensation of the free diamines with the S-methylisothiourea derivative of 5-aminonicotinic acid yielded the corresponding 7-membered guanidine derivative. Further reaction of this guanidine with glycine-beta-amino acid ester followed by hydrolysis and acidification afforded the title compound.

Example 58

Dialkyl / diol substituted seven-membered cyclic guanidine compounds were prepared by the sequence of reactions depicted in the following scheme. From known carbobenzo-

4> 4 4

4

4

184

4 4

4 · 4 44 «44444

4 4

4 xy-protected diamines (J. Med. Chem. 1996, 39, 2156) were synthesized by reacting the 7-membered cyclic guanidines by reacting the free diamine precursor with the S-methylisothiourea derivative of 5-aminonicotinic acid. Condensation of the resulting guanidine-carboxylic acid with glycine-beta-aminoester followed by hydrolysis and acidification gave the final desired compound.

Scheme 3

Z ^is benzyloxycarbonyl

R = CH ₃ , CH / M CH ₂ Ph, CF ₃ , CH ₂ F, alkyl

Example 59

The dihydroxy compounds were prepared by cleaving the cyclic ketal precursor under acidic guanzyases as shown in the following scheme.

H hn ^ V ^N o

of

COOH c

X-Y = halide

185

· ♦ ·· ·· • fe • « • ·· • • fefe • • • • • • • • « • · ♦ · ♦ «· • «· • 9 « • fe • fe • · • fe • • fe • «

Example 60

Compounds containing a seven-membered diol substituted with cyclic guanidines from the starting D or meso-vine derivatives are prepared according to the above method.

The activity of the compounds of the invention was tested according to the following measurements. The test results from these measurements are shown in Table 1.

Vitronectin adhesion measurement

Material

The human vitronectin receptor (α _ν β ₃ ) was purified from the human placenta as described previously [Pytela et al., Methods in Enzymology, 144: 475-489 (1987)]. Human vitronectin was purified from freshly frozen plasma as previously described [Yatohgo et al., Cell Structure and Function, 13: 281-292 (1988)]. Biotinylated human vitronectin was prepared by NHS-biotin compounds from Pierce Chemical Company (Rockford, IL) with purified vitronectin as previously described [Charo et al., J. Biol. Chem., 266 (3): 1415-1421 (1991)]. The buffer used for measurement, OPD substrate tablets and BSA for RIA measurements were obtained from Sigma (St. Louis, MO). An anti-biotin antibody was obtained from Calbiochem (La Jolla, CA). Linbro microtiter plates were obtained from Flow Labs (MsLean, VA). ADP reagents were obtained from Sigma (St. Louis, MO).

Methods

Solid phase receptor measurement

This measurement is entirely consistent with the previously described [Niiya et al., Blood, 70: 475-483 (1987)]. The purified human vitronectin receptor (α _ν β ₃ ) was diluted from the concentrated solution to 1.0 µg / ml in saline containing Tris buffer containing 1.0 mM Ca ^{++ I} Mg ⁺⁺ and Mn ^{++ 1} pH 7, 4 (TBS ⁺⁺⁺ ). Diluted receptor was immediately transferred to Linbro microtiter plates at ΙΟΟμΙ / well (100 ng receptor / well). The plates were sealed, incubated overnight at 4 ° C to bind the receptor to the wells. All subsequent steps were performed at room temperature. Plates were emptied and 200 μ11% BSA was added to the plastic for RIA measurements in TBS ⁺⁺⁺ (TBS ⁺⁺⁺ / BSA). After a 2 hour incubation, the plates were washed with TBS ⁺⁺⁺ using a 96 well plate washer. From this this to this to it to it to it this to this this to it

A 2 mM dilution concentration generated a logatithmic dilution series using 2 nM biotinylated vitronectin in TBS ⁺⁺⁺ / BSA as the diluent. Initial mixing of the labeled ligand with the test (or control) ligand and subsequent transfer of 50 μΙ aliquots to the plates was performed using a CETUS Propette robot; the final concentration of labeled ligand was 1 nM and the highest concentration of test compound was 1.0 10 ^-4 M. The competition again occurred two hours after washing the wells with a plate washer. 125 μΙ of affinity purified horseradish peroxidase labeled with goat anti-biotin antibody diluted 1: 3000 in TBS ⁺⁺⁺ / BSA was added to each well. After 30 minutes, the plates were washed and incubated with OPD / H 2 O 2 substrate in 100 mM / Ι citrate buffer, pH 5.0. The plate was read on a microtiter plate reader at 450 nm and the final A450 was read at the time the control wells reached an absorbance of approximately 1.0. Data were analyzed using an EXCEL J spreadsheet. Mean, standard deviation, and% CV were determined for twice measured concentrations. Mean A450 values were normalized to the average of four maximum binding controls (no competitor added) (B-MAX). Normalized values were used in a four-parameter curve algorithm [Rodbard et al., Int. Atomic Energy Agency, Vienna, pp. 469 (1977)] is plotted on a semi-logarithmic scale, and the computed concentration corresponding to 50% inhibition of the maximum binding of biotinylated vitronectin (IC50) and corresponding ^R2 was reported for those compounds exhibiting greater than 50% inhibition at highest concentration tested; otherwise, the IC50 is reported to be greater than the highest concentration tested. Β - [[2 - [[5 [(aminoiminomethyl) amino] -1-oxopentyl] amino] -1-oxoethyl] amino] -3-pyridine-propanoic acid was included as a positive control in each plate [USSN 08/375, 338, Example 1], which is a potent α 1β 3 antagonist (IC _{5 in the} range of 3 to 10 nM).

Receptor measurement of purified iib / iiia

Material

Human fibrinogen receptor (α _ν β3) was purified from dead platelets (Pytela, R., Pierschbacher, MD, Argraves, S., Suzuki, S., and Rouslahti, E. "Arginine-GlycineAspartic acid adhesion receptors", Methods in Enzymology 144 (1987) 475-489). Human vitronectin was purified from freshly frozen plasma as described in Yatohgo, T., Izumi, M., Kashiwagi, H., and Hayashi, M., "Novel purification of vitronectin from human." · «« ··· ··· ···· · · · · · · · · ·

487 ;:::: ··: · ·:::

Plasma by heparin affinity chromatography, Cell Structure and Function 13 (1988): 281-292. Biotinylated human vitronectin was prepared by combining NHS-biotin from Pierce Chemical Company (Rockford, IL) with purified vitronectin as described previously. (Charo, IF, Nannizzi, L., Phillips, DR, Hsu, MA, Scarborough, RM, "Inhibition of fibrinogen binding to GP IIb / IIIa by GP IIa peptide", J. Biol. Chem. 266 (3) ( 1991): 1415-1421. The buffer used for measurement, OPD substrate tablets and BSA for RIA measurements were obtained from Sigma (St. Louis, MO). Anti-biotin antibodies were obtained from Calbiochem (La Jolla, CA). Linbro microtiter plates were obtained from Flow Labs (McLean, VA). The ADP reagent was obtained from Sigma (St. Louis, MO).

Methods

Measurement of solid phase receptors

This measurement is consistent with Niiya, K., Hodson, E., Bader, R., ByersWard, V. Koziol, JA, Plow, E, F. and Ruggeri, ZM, "Increased surface expression of the 11b / 11la glycoprotein membrane complex induced by platelet activation: relationships to the binding of fibrinogen and platelet aggregation", Blood 70 (1987): 475-483. Purified human fibrinigenic receptor (α _ν β ₃ ) was diluted from the stock solution to 1.0 gg / ml in saline containing Tris buffer and 1.0 mM Ca ⁺⁺ , Mg ⁺⁺ and Mn ⁺⁺ ions, pH (TBS ^{+ ++} ). The diluted receptor was immediately transferred to 100 μbro / well (100 ng receptor / well) in Linbro microtiter plates. The plate was sealed and incubated overnight at 4 ° C to bind the receptor to the wells. All remaining steps were carried out at room temperature. The measured plates were emptied and 200 μ11% BSA for TIA ⁺⁺⁺ (TBS ⁺⁺⁺ / BSA) was added to the plastic coated block. After 2 hours incubation, the measured plates were washed with TBS ⁺⁺⁺ using a 96 well plate washer. A logarithmic series of measurements of the test compound and controls was performed from an initial concentration of 2 mM and using 2 nM biotinylated vitronectin in TBS ⁺⁺⁺ / BSA diluent. This initial mixing of the labeled ligand with the test (control) ligand and the subsequent application of 50 μΙ aliquots into the assay plate was performed using a CETUS Propette robot; the final concentration of labeled ligand was 1 nM and the highest concentration of test compound was 1.0 x 10 ^-4 M. After competition for two hours, all wells were washed with a washer as above. Horseradish peroxidase purified affinity goat anti-biotin antibody was diluted 1: 3000 in TBS ⁺⁺⁺ / BSA and 125 μΙ was added to each well. After 30 minutes, the plates were washed and

··

188 ··· *

were incubated with ODD / H ₂ O ₂ substrate in 100 mM citrate buffer, pH 5.0. Plates were evaluated in a microtiter plate reader at 450 nm and at the maximum binding control, when the absorbance of the wells was approximately 1.0, the final grade was used for analysis. Data were analyzed using an EXCEL J spreadsheet. Mean, standard deviation, and% CV were determined for twice measured concentrations. Mean values As normalized to the mean of the four maximum binding controls (no competitor added) (BMAX). Normalized values were used in a four-parameter curve algorithm [Rodbard et al., Int. Atomic Energy Agencv, Vienna, pp. 469 (1977)] is plotted on a semi-logarithmic scale, and the computed concentration corresponding to 50% inhibition of the maximum binding of biotinylated vitronectin (IC50) and corresponding ^R2 was reported for those compounds exhibiting greater than 50% inhibition at highest concentration tested; otherwise, the IC50 is reported to be greater than the highest concentration tested. P - [[2 - [[5 - [(aminoiminomethyl) amino] -1-oxopentyl] amino] -1-oxoethyl] amino] -3-pyridinepropanoic acid was included as a positive control in each plate [USSN 08/375, 338 , Example 1], which is a potent α _ν β ₃ antagonist (IC 50 in the range of 3 to 10 nM).

Measurement of platelet-rich human plasma

Healthy donors without aspirin were selected from the group of volunteers.

Platelet-rich plasma collection and subsequent ADP-induced platelet clotting measurements were performed as described in Zucker, Μ. B., &quot; Platelet Aggregation Measured by the Photometric Method &quot;, Methods in Enzymology 169 (1989): 117-133. Standard venipuncture techniques were used, using a throttle valve to draw 45 ml of whole blood into a 60 ml syringe containing 5 ml of 3.8% trisodium citrate. After vigorous stirring in the syringe, the clotted blood was transferred to a 50 ml conical polyethylene tube. The blood was centrifuged for 12 minutes at room temperature and 200 xg speed to sediment other cells except platelets. Platelet-rich plasma was transferred to a polyethylene tube and stored at room temperature until further use. Platelet-free plasma was obtained from another 15 minute centrifugation at 2000 xg. The number of platelets normally ranges from 300,000 to 500,000 per microliter. Platelet-rich plasma (0.45 ml) was aliquoted into silicone cuvettes and stirred for 1 minute (1100 rpm) at 37 ° C before adding 50 μ 50 of diluted test compound. After stirring for 1 minute (C • · · ··· «

4 4 4 4

189j j ·; ί ···· i:

• · ·· 44 4 49 4 induced aggregation by adding 50 μΙ 200 μΜ ADP. Aggregation was carried out for 3 minutes in a Payton two-channel aggregometer (Payton Scientific, Buffalo, NY). To determine the dose-response curve, the percent inhibition at maximum response (with saline control) was used for the test compound dilution series. All compounds were tested in duplicate and the concentration at half maximal inhibition (IC 50) was calculated graphically according to a dose-response curve for compounds showing 50% or greater inhibition at the highest concentration tested; otherwise, the IC50 is reported to be greater than the highest concentration tested.

1.90

Example 6. «Νβ ₃ (IC», nm) IIIb / IIIa (IC », nm) 1 1.58 313 2 1.32 272 3 0.41 298 4 0.74 581 5 0.42 884 6 0.27 1260 7 0.21 361 8 95.1 857 9 0.18 244 10 0.14 161 11 0.34 462 12 4.5 442 13 9.32 780 14 12.4 772 15 Dec 25.4 1040 16 22000 6180 17 0.46 724 18 0.43 798 19 Dec 2.96 2490 20 May 0.32 2020 21 0.2 551 22nd 6.96 1710 23 3600 10100 24 91200 15800 25 1870 3910 26 0.71 831 27 Mar: 0.26 482 28 0.43 '984 29 0.46 1385 30 155 29.5 31 3942 1803

* 21 '

Example # drawbar (IC ™, nm) IIIb / IIIa (IC _M , nm) 32 2.1 49 33 0.2 520 34 0.34 1203 35 0.14 340 36 1.13 970 37 63.8 267 38 278 115 39 1917 2458 40 1.5 20630 41 10.2 35400 42 3.1 5954 43 6.5 7527 44 42.1 1061 45 1094 1678 46 1.97 2693 47 0.99 6709 48 4.3 27910

py 2t> / 0 ~ j / 7-p

C J. ý / r.

192

pudoJM IW * Ie # '

Claims (22)

Claims Heterocyclic glycyl-beta-alanine derivative of the general formula I N-x # Uh ·%: W * ¹ -, 1 '(CH 2), O> -c (1), where HET is a 5 to 8 membered monocyclic heterocycle, optionally unsaturated, containing 1 to 4 heteroatoms selected from the group consisting of O, N, or S; wherein X ¹ is selected from the group consisting of CH, CH ₂ , N, NH, O and S; And is N-R ' 193 Y ¹ wherein Υ ¹ is selected from the group consisting of NR ² , O, and S; R ² is selected from the group consisting of H; alkyl; aryl; hydroxy; alkoxy; cyano; nitro; amino; alkenyl; alkynyl; amido; alkylcarbonyl; arylcarbonyl; alkoxycarbonyl; aryloxycarbonyl; haloalkylcarbonyl; haloalkoxycarbonyl; alkylthiocarbonyl; arylthiocarbonyl; acyloxymethoxycarbonyl; alkyl optionally substituted with one or more substituents selected from the group consisting of lower alkyl, halide, hydroxyl, haloalkyl, cyano, nitro, carboxyl, amino, alkoxy, aryl or aryl optionally substituted with one or more halo, haloalkyl, lower alkyl, alkoxy, cyano , alkylsulfonyl, alkylthio, nitro, carboxy, amino, hydroxyls, sulfuric acid, sulfonamides, aryls, fused aryl, monocyclic heterocycles or fused monocyclic heterocycles; aryl optionally substituted with one or more substituents selected from the group consisting of halide, haloalkyl, hydroxy, lower alkyl, alkoxy, methylenedioxy, ethylenedioxy, cyano, nitro, alkylthio, alkylsulfonyl, sulfuric acid, sulfonamide, carboxyl derivatives, amino, aryl, fused aryl, monocyclic heterocycles and fused monocyclic heterocycles; monocyclic heterocycles and monocyclic heterocycles optionally substituted with one or more substituents selected from the group consisting of halide, haloalkyl, lower alkyl, alkoxy, amino, nitro, hydroxy, carboxyl derivatives, cyano, alkylthio, alkylsulfonyl, sulfuric acid, sulfonamide, aryl, or fused aryl; or R ² together form R ^{7 a} 4 to 12 membered heterocycle containing two nitrogen atoms optionally substituted with one or more substituents selected from the group consisting of lower alkyl, thioalkyl, alkylamino, hydroxy, keto, alkoxy, halogen, phenyl, amino, carboxyl or carboxylic ester , spiro dioxolane and condensed phenyl; 194 ······· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·· · ·· · · · R ² together with R ⁷ forms a 4 to 12 membered heterocycle, optionally unsaturated, containing one or more heteroatoms selected from the group consisting of O, N and S; R ² together with R ⁷ forms a 5- to 9-membered heterocycle, optionally substituted with one or more substituents selected from the group consisting of lower alkyl, phenyl, alkoxy and hydroxy; R ² together with R ⁷ forms a 5 membered heterocycle, fused with an aryl or heterocycle; R ⁷ , when not associated with R ² , and R ® are independently selected from the group consisting of H; alkyl; alkenyl; alkynyl; aralkyl; amino; alkylamino; hydroxy; alkoxy; arylamino; amido; alkylcarbonyl; arylcarbonyl; alkoxycarbonyl; aryloxy; aryloxycarbonyl; haloalkylcarbonyl; haloalkoxycarbonyl; alkylthiocarbonyl; arylthiocarbonyl; acyloxymethoxycarbonyl; cycloalkyl; bicycloalkyl; aryl; acyl; benzoyl; alkyl optionally substituted with one or more substituents selected from the group consisting of lower alkyl, halide, hydroxy, haloalkyl, cyano, nitro, carboxyl derivatives, amino, alkoxy, thio, alkylthio, sulfonyl, aryl, aralkyl, aryl optionally substituted with one or more substituents selected from the group consisting of halide, haloalkyl, lower alkyl, alkoxy, methylenedioxy, ethylenedioxy, alkylthio, haloalkylthio, thio, hydroxy, cyano, nitro, carboxyl derivatives, aryloxy, amido, acylamino, amino, alkylamino, dialkylamino, trifluoroalkoxy, trifluoromethyl, sulfonyl, alkylsulfonyl, haloalkylsulfonyl, sulfuric acid, sulfonamide, aryl, fused aryl, monocyclic heterocycles, fused monocyclic heterocycles; monocyclic heterocycles; monocyclic heterocycles optionally substituted with one or more substituents selected from the group consisting of halide, haloalkyl, lower alkyl, alkoxy, methylenedioxy, ethylenedioxy, alkylthio, haloalkylthio, thio, hydroxy, cyano, nitro, carboxyl derivatives, aryloxy, amido, acylamino, amino, alkylamino, dialkylamino, trifluoroalkoxy, trifluoromethylsulfonyl, alkylsulfonyl, sulfuric acid, sulfonylamide, aryl, fused aryl, monocyclic heterocycles or fused monocyclic heterocycles; monocyclic heterocycles optionally substituted with one or more substituents selected from the group consisting of halide, haloalkyl, lower 195 alkyl, alkoxy, aryloxy, amino, nitro, hydroxy, carbonyl derivatives, cyano, alkylthio, alkylsulfonyl, aryl, fused aryl; monocyclic and bicyclic heterocycloalkyls; -SO ₂ R ¹⁰ , wherein R ¹⁰ is selected from the group consisting of alkyl, aryl and monocyclic heterocycles, all optionally substituted with one or more substituents selected from the group consisting of halides, haloalkyl, alkyl, alkoxy, cyano, nitro, amino, acylamino, trifluoroalkyl, amido, alkylamino sulfonyl, alkylsulfonyl, alkylsulfonylamine, alkylamine, dialkylamine, trifluoromethylthio, trifluoralkoxy, trifluoromethylsulfonyl, aryl, aryloxy, thio, alkylthio, and monocyclic heterocycles; and O II wherein R ¹⁰ is as defined above; NR ⁷ and R ⁸ together form a 4 to 12 membered nitrogen-containing monocyclic or bicyclic ring optionally substituted with one or more substituents selected from the group consisting of lower alkyl, carboxyl derivatives, aryl or hydroxy, and wherein the monocyclic or bicyclic ring optionally contains a heteroatom selected from a group consisting of O, N and S; R ⁵ is selected from the group consisting of H, alkyl, alkenyl, alkynyl, benzyl, and phenethyl; or A is Ý ² 196 wherein Y ² is selected from the group consisting of alkyl; cycloalkyl; bicycloalkyl; aryl; monocyclic heterocycles; alkyl optionally substituted with aryl, which may also be optionally substituted with one or more substituents selected from the group consisting of halo, haloalkyl, alkyl, nitro, hydroxy, alkoxy, aryloxy, aryl or fused aryl; aryl optionally substituted with one or more substituents selected from the group consisting of halogen, haloalkyl, hydroxy, alkoxy, aryloxy, aryl, fused aryl, nitro, methylenedioxy, ethylenedioxy or alkyl; alkynyl; alkenyl; -SR ⁹ and OR ⁹ , wherein R ⁹ is selected from the group consisting of H; alkyl; aralkyl; aryl; alkenyl; and alkynyl; or R ⁹ together with R ⁷ forms a 4-12 membered heterocycle containing one nitrogen, sulfur or oxygen atom, optionally substituted with lower alkyl, hydroxy, keto, phenyl, carboxyl or carboxylic ester and fused phenyl; or R ⁹ together with R ⁷ forms a thiazole; oxazole; benzoxazole; or benzothiazole; and R ⁵ and R ⁷ are as defined above; Y ² , when Y ^{2 is} carbon, together with R ⁷ forms a 4-12 membered ring containing one or two nitrogen atoms, optionally substituted with alkyl, aryl, keto or hydroxy; And is NR ² N-R ⁷ Jo where R ² and R ⁷ form together 5-8-membered heterocycle containing two nitrogen atoms, which is optionally substituted with one or more substituents selected from the group consisting of lower alkyl, hydroxy, alkoxy, keto, phenyl, or carboxyl derivatives; and R ⁸ is selected from the group consisting of alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkylcarbonyl, haloalkoxycarbonyl, alkylthiocarbonyl, arylthiocarbonyl or acyloxymethoxycarbonyl; and R ⁵ is as defined above R ² and R ⁷ together form a membered heterocycle such as imidazole or pyrimidone; or A is R ⁵ NR ² -N = C ₇ NR ⁷ R ⁸ wherein R ² and R ⁷ together form a 5-8 membered heterocycle containing two nitrogen atoms, which is optionally substituted with a hydroxy, keto, phenyl or alkyl group; and R ⁸ are both selected from the group consisting of alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkylcarbonyl, haloalkoxycarbonyl, alkylthiocarbonyl, arylthiocarbonyl and acyloxymethoxycarbonyl; Z ¹ is one or more substituents selected from the group consisting of H; alkyl; hydroxy; alkoxy; aryloxy; a halide; haloalkyl; haloalkoxy; nitro; amino; alkylamino; acylamino; dialkylamino; cyano; alkylthio; alkylsulfonyl; carboxyl derivatives; trihaloacetamide; acetamide; acyl; aryl; fused aryl; cycloalkyl; thio; monocyclic heterocycles; fused monocyclic heterocycles; and A, wherein A is as defined above; • ·· · ·· 1QO ······· · · · · IVO ·· ·· ·· · ·· ··· V is selected from the group consisting of -N- (R ⁶ ) -, wherein R ⁶ is selected from the group consisting of H; lower alkyl; cycloalkyl; aralkyl; aryl; and monocyclic heterocycles; or R ⁶ together with Y forms a 4 to 12 membered ring containing one nitrogen atom; Y, Y ³ , Z and Z ³ are independently selected from the group consisting of hydrogen; alkyl; aryl; and cycloalkyl; or Y and Z together form a cycloalkyl; or Y ³ and Z ³ together form a cycloalkyl; n is an integer, 1, 2 or 3; t is an integer, 0, 1 or 2; p is an integer, 0, 1, 2 or 3; R is XR ³ wherein X is selected from the group consisting of O, S and NR ⁴ wherein R ³ and R ⁴ are independently selected from the group consisting of hydrogen; alkyl; alkenyl; alkynyl; haloalkyl; aryl; arylalkyl; sugars; steroids; polyalkyl ethers; alkylamido; alkyl N, N-dialkylamido; pivaloyloxymethyl; and in the case of the free acid, all pharmaceutically acceptable salts thereof; R ¹ is selected from the group consisting of hydrogen, alkyl, alkenyl; alkynyl; aryl; carboxyl derivatives; haloalkyls; cycloalkyls; monocyclic heterocycles; monocyclic heterocycles optionally substituted with alkyl, halide, haloalkyl, cyano, hydroxy, aryl, fused aryl, nitro, alkoxy, aryloxy, alkylsulfonyl, arylsulfonyl; sulfonamide; thio, alkylthio, carboxyl derivatives, amino, amido; alkyl is optionally substituted by one or more of halide, haloalkyl, hydroxy, alkoxy, aryloxy, thio, alkylthio, alkynyl, alkenyl, alkyl, arylthio, alkylsulfoxide, alkylsulfonyl, arylsulfoxide, arylsulfonyl, cyano, nitro, amino, alkylamino, dialkylamino, alkylsulfonamide, arylsulfonamide, acylamide, carboxyl derivatives, sulfonamide, sulfuric acid, phosphorous acid derivatives, phosphorous acid derivatives, aryl, arylthio, arylsulfoxide or arylsulfone, all of which are optionally substituted on the aryl ring with halide, alkyl, haloalkyl, hydroxy, cyano, nitro carboxyl derivatives, alkoxy, aryloxy, amino, alkylamino, 199 • 4 · · · ······ · ·> ······ · Dialkylamino, amido, aryl, fused aryl, monocyclic heterocycles, and fused monocyclic heterocycles, monocyclic heterocyclothio, monocyclic heterocyclosulfoxide and monocyclic heterocyclic sulfone, which may optionally be substituted with halide, haloalkyl, nitro, hydroxy, alkoxy, fused aryl or alkyl; alkylcarbonyl, haloalkylcarbonyl and arylcarbonyl; aryl may be optionally substituted at one or more positions by halogen, haloalkyl, alkyl, alkoxy, aryloxy, methylenedioxy, ethylenedioxy, alkylthio, haloalkylthio, thio, hydroxy, cyano, nitro, acyloxy, carboxyl derivatives, carboxyalkoxy; amido, acylamino, amino, alkylamino, dialkylamino, trifluoroalkoxy, trifluoromethylsulfonyl, alkylsulfonyl, sulfuric acid, sulfonamide, aryl, fused aryl, monocyclic heterocycles and fused monocyclic hetrocycles; and About CR ⁷ Wherein R ⁷ and R ⁸ are as defined above and together with the nitrogen, R ⁷ and R ⁸ include an amino acid; and R ¹¹ is selected from the group consisting of H, alkyl, aralkyl, alkenyl, alkynyl, haloalkyl, or haloalkynyl, or R ¹¹ together with Y forms a 4 to 12 membered ring containing one nitrogen atom; or a pharmaceutically acceptable salt thereof. * · A heterocyclic glycyl-beta-alanine derivative according to claim 1 of the general formula II 200 wherein R ³² is H, alkyl, alkoxyalkyl, aminoalkyl, dialkylaminoalkyl, wherein the alkyl group is optionally substituted with one or more substituents selected from the group consisting of hydroxy , alkoxy, amino, alkylamino, dialkylamino, aryl or alkylkylsulfonyl, carboxyl and carboxyl derivatives. The heterocyclic glycyl-beta-alanine derivative according to claim 2, selected from the group consisting of 201 • · · · · · · · · · φ · φ · φ · φ · Βγ ·· · · ·· ·· * 207 • a 4 · a and a 4. A compound according to claim 1 wherein (II)! Het ï _ie \ xt J ^e As The heterocyclic glycyl-beta-alanine derivative according to claim 4, selected from the group consisting of: 208 • 0 · 0 0 · 0 0 · 0 209 • · ·· ·· φφφφ ·· 211 ΦΦ φ φ · φ φ · · · · · · · φ φ φ φ φ 213 ·· · AA A Heterocyclic glycyl-beta-alanine derivative according to claim 1 of the general formula II (II) 214 ΦΦ · kde φ kde kde kde kde kde kde kde kde φ kde kde kde kde The heterocyclic glycyl-beta-alanine derivative according to claim 6 selected from the group consisting of 9999 «· 9 * 9 · 9 9 9 9 9 9 999 • fefe · fe 8. A compound according to claim 1 of formula II 216 • fefe · fe · fe • fe (II), i Het i, τΧ '' is 9. A compound according to claim 8 having the formula: &lt; tb &gt; ______________________________________ &lt; tb &gt; 10. A compound according to claim 1 of formula II (CHA) wherein: Het &lt; 1 &gt; s &lt; + &gt;, R &lt; 11 &gt; (II), ^X &lt; 1 &gt; Het &lt; 1 &gt; λ / is N ^ S • 9 A heterocyclic glycyl-beta-alanine derivative according to claim 10 of the formula 218 «· · NH 12. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 and a pharmaceutically acceptable carrier. A method for the treatment of conditions induced by integrin α _ν β ₃ in a mammal in need thereof, comprising administering a therapeutically effective inhibitory amount of α _ν β _{3 of the} compound of claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 . The method of claim 13, wherein the condition being treated is tumor metastasis. The method of claim 13, wherein the condition to be treated is solid tumor growth. The method of claim 13, wherein the condition to be treated is angiogenesis. The method of claim 13, wherein the condition to be treated is osteoporosis. The method of claim 13, wherein the condition to be treated is humoral hypercalcaemia or malignancy. The method of claim 13, wherein the condition being treated is smooth muscle cell migration. 0 0 219 0 00 00 0 • · · 0 0 0 0 0 0 0 0 0 00 000 0000 0 · 0 0 0 0 The method of claim 13, wherein restenosis is inhibited. The method of claim 13, wherein the condition to be treated is rheumatoid arthritis. The method of claim 13, wherein the condition to be treated is macular degeneration.