KR20010022413A - Benzyl compounds which inhibit leukocyte adhesion mediated by VLA-4 - Google Patents

Abstract

The present invention describes compounds that bind to VLA-4. In addition, these specific compounds prevent leukocyte adhesion, in particular leukocyte adhesion mediated by VLA-4. Such compounds are used in the treatment of inflammatory diseases in mammals such as humans, such as asthma, Alzheimer's disease, atherosclerosis, AIDS dementia, diabetes, inflammatory bowel disease, rheumatoid arthritis, tissue transplantation, tumor metastasis and myocardial ischemia. Useful. The compounds may also be administered to treat inflammatory brain diseases such as multiple sclerosis.

Description

Benzyl compounds which inhibit leukocyte adhesion mediated by VLA-4}

Background of the Invention

Field of invention

The present invention relates to compounds which inhibit leukocyte adhesion, in particular leukocyte adhesion mediated by VLA-4.

Reference

The following publications, patents, and patent applications are cited herein as superscript numbers:

¹ European Patent Publication No. 330,506 by Hemler and Takada (published date: 30/98/98)

² Elices, et al., Cell, 60: 577-584 (1990)

³ Springer, Nature, 346: 425-434 (1990)

⁴ Osborn, Cell, 62: 3-6 (1990)

⁵ Vedder, et al., Surgery, 106: 509 (1989)

⁶ Pretolani, et al., J. Exp. Med., 180: 795 (1994)

⁷ Abraham, et al., J. Clin. Invest., 93: 776 (1994)

⁸ Mulligan, et al., J. Immunology, 150: 2407 (1993)

⁹ Cybulsky, et al., Science, 251: 788 (1991)

¹⁰ Li, et al., Arterioscler. Thromb., 13: 197 (1993)

¹¹ Sasseville, et al., Am. J. Path., 144: 27 (1994)

¹² Yang, et al., Proc. Nat. Acad. Science (USA), 90: 10494 (1993)

¹³ Burkly et al., Diabetes, 43: 529 (1994)

¹⁴ Baron, et al., J. Clin. Invest., 93: 1700 (1994)

¹⁵ Hamann, et al., J. Immunology, 152: 3238 (1994)

¹⁶ Yednock, et al., Nature, 356: 63 (1992)

¹⁷ Baron, et al., J. Exp. Med., 177: 57 (1993)

¹⁸ van Dinther-Janssen, et al., J. Immunology, 147: 4207 (1991)

¹⁹ van Dinther- Janssen, et al., Annals. Rheumatic Dis., 52: 672 (1993)

²⁰ Elices, et al., J. Clin. Invest., 93: 405 (1994)

²¹ Postigo, et al., J. Clin. Invest., 89: 1445 (1991)

²² Paul, et al., Transpl. Proceed., 25: 813 (1993)

²³ Okahara, et al., Can. Res., 54: 3233 (1994)

²⁴ Paavonen, et al., Int. J. Can., 58: 298 (1994)

²⁵ Schadendorf, et al., J. Path., 170: 429 (1993)

²⁶ Bao, et al., Diff., 52: 239 (1993)

²⁷ Lauri, et al., British J. Cancer, 68: 862 (1993)

²⁸ Kawaguchi, et al., Japanese J. Cancer Res., 83: 1304 (1992)

²⁹ US Pat. No. 5,510,332 to Kogan et al. (1996.4.23.)

³⁰ International Publication No. WO 96/01644

All of the above publications, patents, and patent applications are cited herein in their entirety to the same extent as if each individual publication, patent, and patent application were specifically and individually incorporated by reference in their entirety.

Technical description

VLA-4 (also known as α ₄ β ₁ integrin and CD49d / CD29), first identified by Hemler and Takada ^1, is a group of β ₁ integrins of cell surface receptors that contain two subunits, α chain and β chain, respectively. One member. VLA-4 includes α ₄ chains and β ₁ chains. There are at least nine β ₁ integrins, all of which share the same β ₁ chain and each have a separate α chain. All nine of these receptors bind different complements of various cell matrix molecules such as fibronectin, laminin and collagen. VLA-4 binds, for example, fibronectin. VLA-4 is unique among β ₁ integrins that also bind to non-matrix molecules expressed by endothelial cells and other cells. Such non-matrix molecules include VCAM-1 expressed in umbilical vein endothelial cells of cytokine activated humans in culture. Separate epitopes of VLA-4 contribute to fibronectin and appear to independently inhibit VCAM-1 binding activity and each activity ² .

Intracellular adhesion mediated by VLA-4 and other cell surface receptors is associated with a number of inflammatory responses. At the wound site or other inflammatory stimulus site, activated vascular endothelial cells express molecules that attach to leukocytes. The mechanism of action of leukocyte adhesion to endothelial cells involves, in part, that the cell surface receptors of leukocytes recognize and bind to the corresponding cell surface molecules of endothelial cells. Once bound, white blood cells travel through the walls of blood vessels to enter the wound site, releasing chemical media that fight infection. For attachment receptors of the immune system, see, for example, Springer ³ and Osborn ⁴ .

Inflammatory brain diseases such as experimental autoimmune encephalomyelitis (EAE), multiple sclerosis (MS) and meningitis are examples of central nervous system diseases in which endothelial / leukocyte adhesion mechanisms also destroy healthy brain tissue. Many leukocytes migrate through the blood brain barrier (BBB) in subjects with the inflammatory diseases described above. Leukocytes release toxic mediators that cause a wide range of brain damage causing reduced brain conduction and paralysis.

In other organ systems, tissue damage also occurs through adhesion mechanisms that produce the migration or activation of white blood cells. For example, early damage after cardiac myocardial ischemia can be complicated by the influx of leukocytes into damaged cells, still causing further damage. Vedder et al. ⁵ ]. Other inflammatory diseases mediated by adhesion mechanisms include, for example, asthma ^6-8 , Alzheimer's disease, atherosclerosis ^9-10 , AIDS dementia ¹¹ , diabetes ^12-14 (including acute pediatric diabetes), inflammatory bowel disease ¹⁵ (Including ulcerative colitis and Crohn's disease), multiple sclerosis ^16-17 , rheumatoid arthritis ^18-21 , tissue transplantation ²² , tumor metastasis ^23-28 , meningitis, encephalitis, ataxia and other cerebral trauma, nephritis, retinitis, atopic dermatitis And acute leukocyte mediated lung injury such as occurs in psoriasis, myocardial ischemia, and adult respiratory distress syndrome.

In view of the above, assays for measuring the levels of VLA-4 in biological samples containing VLA-4 are useful, for example, in diagnosing VLA-4 mediated diseases. In addition, despite this advance in understanding leukocyte adhesion, the art has only recently addressed the use of adhesion inhibitors in the treatment of inflammatory brain diseases and other inflammatory diseases ^29,30 . The present invention addresses these and other needs.

Summary of the Invention

The present invention provides compounds that bind to VLA-4. Such compounds can be used, for example, to analyze the presence of VLA-4 in a sample, and inhibit intracellular adhesion mediated by VLA-4 (eg, binding of VCAM-1 to VLA-4). It can be used in pharmaceutical compositions. The compounds of the present invention, which are the compounds of formula I and pharmaceutically acceptable salts thereof, have a binding affinity for VLA-4 represented by an IC ₅₀ of less than about 15 μM.

In Formula I above,

R ¹ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclic, substituted heterocyclic, heteroaryl and substituted heteroaryl,

R ² and R ³ may form a saturated heterocyclic group or a saturated substituted heterocyclic group together with a nitrogen atom bonded to R ² and a carbon atom bonded to R ³ , provided that when mono-substituted, Substituents in such saturated heterocyclic groups are not carboxy,

R ⁵ is a - is selected from the group consisting of heteroaryl (wherein, n is an integer from 1 to _{4), - (CH 2)} n - aryl and - (CH _{2) n}

Q is —C (X) NR ⁷ —, where R ⁷ is selected from the group consisting of hydrogen and alkyl, X is selected from the group consisting of oxygen and sulfur,

Provided that when R ¹ is 2,4,6-trimethylphenyl, R ² and R ³ together with the pendant nitrogen and carbon atoms form a pyrrolidinyl ring and Q is -C (O) NH-, then R ⁵ Is not benzyl,

When R ¹ is p-methylphenyl and R ² and R ^{3 together} with the pendant nitrogen and carbon atoms form a pyrrolidinyl ring derived from D-proline, and Q is -C (O) NH-, ⁵ is not benzyl derived from D-phenylalanine.

In another embodiment, the compounds of the present invention are also provided as prodrugs that are converted in vivo to compounds of formula I above (eg, hydrolysis, metabolism, etc.). A preferred example of this embodiment is to modify the carboxylic acid of the compound of formula (I) into a group which is converted in vivo to a carboxylic acid (including salts thereof). In a particularly preferred embodiment, such prodrugs represent compounds of formula (IA) and pharmaceutically acceptable salts thereof.

In Formula IA above,

R ¹ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, heterocyclic, substituted heterocyclic, heteroaryl and substituted heteroaryl,

R ² and R ³ may form a saturated heterocyclic group or a saturated substituted heterocyclic group together with a nitrogen atom bonded to R ² and a carbon atom bonded to R ³ , provided that when mono-substituted, Substituents in such saturated heterocyclic groups are not carboxy,

R ⁵ is a - is selected from the group consisting of heteroaryl (wherein, n is an integer from 1 to _{4), - (CH 2)} n - aryl and - (CH _{2) n}

R ⁶ is amino, alkoxy, substituted alkoxy, cycloalkoxy, substituted cycloalkoxy, -O- (N-succinimidyl), -NH-adamantyl, -O-cholest-5-ene-3-β -Yl, -NHOY, where Y is hydrogen, alkyl, substituted alkyl, aryl and substituted aryl, -NH (CH) _P COOY, where p is an integer from 1 to 8 and Y is as defined above Equals), -OCH ₂ NR ⁹ R ¹⁰ , wherein R ⁹ is selected from the group consisting of -C (O) -aryl and -C (O) -substituted aryl, R ¹⁰ is hydrogen and -CH ₂ COOR ¹¹ (Wherein R ¹¹ is alkyl) and -NHSO ₂ Z, wherein Z is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, heterocyclic and substituted heterocyclic), and

Q is —C (X) NR ⁷ —, where R ⁷ is selected from the group consisting of hydrogen and alkyl, X is selected from the group consisting of oxygen and sulfur,

A: R ¹ is 4-methylphenyl, R ² and R ³ together with the nitrogen and carbon atoms form a pyrrolidin-2-yl ring, R ⁵ is benzyl and Q is -C (O) NH When-R ⁶ is not -NH (CH ₂ ) ₂ CO ₂ Et or-(1R, 2S, 5R)-(-)-methyl ester,

B: R ¹ is 4-methylphenyl, R ² and R ³ together with pendant nitrogen atoms and carbon atoms form a 3-β-phenyl-ring derived from D-proline, R ⁵ is benzyl, Q is- In case of C (O) NH-, R ⁶ is not -OCH ₂ CH ₃ ,

C: R ¹ is 1-N-methyl-3-methyl-5-chloropyrazol-4-yl, and R ² and R ³ together with the pendant nitrogen and carbon atoms form a pyrrolidin-2-yl ring When R ⁵ is benzyl and Q is -C (O) NH-, R ⁶ is not -OCH ₃ ;

D: R ¹ is 4-methylphenyl, R ² and R ³ together with the pendant nitrogen atom and carbon atom form a pyrrolidin-2-yl ring, R ⁵ is D-benzyl and Q is -C (O ) NH-, R ⁶ is not -OCH ₂ CH ₃ ,

E: R ¹ is 4-methylphenyl, R ² and R ³ together with the pendant nitrogen and carbon atoms form a 5,5-dimethyl-1,1-dioxo-thiaprolyl ring, R ⁵ is benzyl , When Q is -C (O) NH-, R ⁶ is not -OC (CH ₃ ) ₃ ,

F: R ¹ is 4-methylphenyl, R ² and R ³ together with pendant nitrogen and carbon atoms form a pyrrolidin-2-yl ring from D-proline, R ⁵ benzyl derived from D-phenylalanine And when Q is -C (O) NH-, R ⁶ is not -OCH ₃ ,

G: R ¹ is n-butyl, R ² and R ³ together with the pendant nitrogen atom and carbon atom form a pyrrolidin-2-yl ring, R ⁵ is benzyl and Q is -C (O) NH If-R ⁶ is not -O-benzyl.

Preferably, in the above compounds of Formula I and Formula IA, R ¹ is a group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heterocyclic, substituted heterocyclic, heteroaryl and substituted heteroaryl Is selected from. More preferably, R ¹ is 4-methylphenyl, methyl, benzyl, n-butyl, 4-chlorophenyl, 1-naphthyl, 2-naphthyl, 4-methoxyphenyl, phenyl, 2,4,6-tri Methylphenyl, 2- (methoxycarbonyl) phenyl, 2-carboxyphenyl, 3,5-dichlorophenyl, 4-trifluoromethylphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl, 4- (CH ₃ C (O) NH-) phenyl, 4-trifluoromethoxyphenyl, 4-cyanophenyl, isopropyl, 3,5-di- (trifluoromethyl) phenyl, 4-t-butylphenyl, 4- t-butoxyphenyl, 4-nitrophenyl, 2-thienyl, 1-N-methyl-3-methyl-5-chloropyrazol-4-yl, phenethyl, 1-N-methylimidazole-4- 1, 4-bromophenyl, 4-aminophenyl, 4-methylamidinophenyl, 4- [CH ₃ SC (= NH)] phenyl, 5-chloro-2-thienyl, 2,5-dichloro-4- Thienyl, 1-N-methyl-4-pyrazolyl, 2-thiazolyl, 5-methyl-1,3,4-thiadiazol-2-yl, 4- [H ₂ NC (S)] phenyl, 4 -Aminophenyl, 4-fluorophenyl, 2-fluorophenyl, 3-fluorophenyl, 3,5-difluorophenyl, pyridine-3- 1, pyrimidin-2-yl, 4- (3'-dimethylamino-n-propoxy) -phenyl.

In one preferred embodiment, R ² and R ³ ring one or two hetero atoms selected from nitrogen, oxygen and sulfur together with the nitrogen atom bonded to the R ² substituent and the carbon atom bonded to the R ³ substituent To form a heterocyclic group or a substituted heterocyclic group of 4 to 6 ring atoms having at least one or two selected from fluoro, methyl, hydroxy, amino, phenyl, thiophenyl and thiobenzyl 10 to 14 molten rings substituted with 4 substituents or melted on another saturated heterocyclic or cycloalkyl ring, such as a cyclohexyl ring, having 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur in the ring Can provide. Such heterocyclic rings include thiazolidinyl (eg L-thiazolidin-4-yl), piperidinyl (eg L-piperidin-2-yl), piperizinyl (eg L-piperizine- 2-yl), thiomorpholinyl (eg L-thiomorpholin-3-yl), pyrrolidinyl (eg L-pyrrolidin-2-yl); 4-hydroxypyrrolidinyl [eg 4-α- (or β-) hydroxy-L-pyrrolidin-2-yl], 4-fluoropyrrolidinyl [eg 4-α- (or β -) Fluoro-L-pyrrolidin-2-yl], 3-phenylpyrrolidinyl [eg 3-α- (or β-) phenyl-L-pyrrolidin-2-yl], 3-thio Phenylpyrrolidinyl [eg 3-α- (or β-) thiophenyl-L-pyrrolidin-2-yl], 4-aminopyrrolidinyl [eg 4-α- (or β-) amino- L-pyrrolidin-2-yl], 3-methoxypyrrolidinyl [eg 3-α- (or β-) methoxy-L-pyrrolidin-2-yl], 4,4-dimethoxypy Substituted pyrrolidinyl, such as rolidin-2-yl; Substituted piperidinyl, such as 4-N-Cbz-piperidin-2-yl; Substituted thiazolidinyl such as 5,5-dimethylthiazolidin-4-yl; 1,1-dioxo-thiazolidinyl (eg L-1,1-dioxo-thiazolidin-4-yl), substituted 1,1-dioxo-thiazolidinyl (eg L-1, 1-dioxo-5,5-dimethylthiazolidin-4-yl), 1,1-dioxothiomorpholinyl (e.g., L-1,1-dioxo-thiomorpholin-3-yl), etc. It includes. Preferably, such a ring does not include a ring which forms an azetidine ring together with the nitrogen atom having R ² and R ³ bonded to R ² and the carbon atom bonded to R ³ .

Q is preferably -C (O) NH- or -C (S) NH-.

R ⁵ is preferably selected from all possible isomers by substituents of the following groups: benzyl, phenethyl, -CH _2- (3-indolyl), -CH _2- (1-naphthyl), -CH _2- (2-naphthyl), -CH ₂ (2-ethyl-T), -CH ₂ - (3- pyridyl), -CH ₂ - (5- imidazolyl), -CH ₂ -3- (1,2 , 4-triazolyl), -CH ₂ - (2- thiazolyl) and the like.

In the compound of formula (IA), R ⁶ is 2,4-dioxo-tetrahydrofuran-3-yl (3,4-enol), methoxy, ethoxy, iso-propoxy, n-butoxy, t-part Methoxy, cyclopentoxy, neo-pentoxy, 2-α-iso-propyl-4-β-methylcyclohexoxy, 2-β-isopropyl-4-β-methylcyclohexoxy, -NH ₂ , benzyloxy , -NHCH ₂ COOH, -NHCH ₂ CH ₂ COOH, -NH-adamantyl, -NHCH ₂ CH ₂ COOCH ₂ CH ₃ , -NHSO ₂ -p-CH ₃ -ψ, -NHOR ⁸ (where R ⁸ is Hydrogen, methyl, iso-propyl or benzyl), O- (N-succinimidyl), -O-cholest-5-en-3-β-yl, -OCH ₂ -OC (O) C (CH ₃ ) ₃ , -O (CH ₂ ) zNHC (O) W, wherein z is 1 or 2, W is pyrid-3-yl, N-methylpyridyl and N-methyl-1,4-dihydro- Is selected from the group consisting of pyrid-3-yl) and -NR "C (O) -R 'wherein R' is aryl, heteroaryl or heterocyclic, and R" is hydrogen or -CH ₂ C (O Is OCH ₂ CH ₃ ).

Preferred compounds within the scope of formula (I) and formula (IA) above include the following examples:

N- (methanesulfonyl) -L-prolyl-L-phenylalanine, N- (α-toluenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prop Rollyl-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L- (N-methyl) phenylalanine, N- (toluene-4-sulfonyl) -L-pipecolinyl-L-phenylalanine, N- (toluene-4-sulfonyl) -D-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L- (4-hydroxy) prolyl-L-phenylalanine, N- (toluene 4-sulfonyl) -L-prolyl-D, L-homophenylalanine, N- (4-chlorobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (1-naphthalenesulfonyl) -L -Prolyl-L-phenylalanine, N- (2-naphthalenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-methoxybenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-t-butylbenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L- (4-fluoro) prolyl-L-phenylalanine, N- (n -Butanesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine, N- (2-methoxycarbonylbenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (2-carboxybenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl ) -L-thiaprolyl-L-phenylalanine, N- (3,5-dichlorobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-trifluoromethoxybenzenesulfonyl) -L- Prolyl-L-phenylalanine, N- (3,4-dichlorobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -D, L- (3-phenyl) prop Rollyl-L-phenylalanine, N- (3,4-dimethoxybenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-nitrobenzenesulfonyl) -L-prolyl-L-phenylalanine, N -(4-acetamidobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-cyanobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sul Ponyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-β- (1-naphthyl) -L-alanine, N- (toluene-4-sulfonyl ) -L-prolyl-β- (2-naphthyl) -L-alanine, N- (toluene-4-sulfonyl) -L-prolyl-β- (2-thienyl) -L-alanine, N- (isopropanesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-β- (3-pyridyl) -L-alanine, N- ( Toluene-4-sulfonyl) -L- (4-phenylthio) prolyl-L-phenylalanine, N- (toluene-4-sulfonyl)-(4-benzylthio) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L-histidine, N- (toluene-4-sulfonyl) -L- (4-amino) prolyl-L-phenylalanine, N- (toluene- 4-sulfonyl) -L-prolyl-L-phenylalanineamide, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L -Prolyl-L-phenylalanine ethyl ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-methoxyamide, N- (toluene-4-sulfonyl) -L-prolyl -L-phenylalanine N-benzyloxyamide, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N- (toluene-4-sulfonyl) amide, N- (toluene-4-sulfonyl ) -L-Prolyl-L-phenylalanine-β-alanine, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine-N- Idroxyamide, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine isopropyl ester, N- (toluene-4-sulfonyl) -L- (4-hydroxy) prolyl-L -Phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanyl- (N-benzoyl) glycine ethyl ester, N- (toluene-4-sulfonyl) -L- (4- Fluoro) prolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L-thiaprolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L- (5 , 5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine ethyl ester, N- (2- Methoxycarbonylbenzenesulfonyl) -L-prolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L- (3-phenyl) prolyl-L-phenylalanine ethyl ester, N- ( Toluene-4-sulfonyl) -L- (4-methoxy) prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine (1S, 2R, 5S)- (+)-Methyl ester, N- ( Toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-hydroxysuccinimide ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine 2- (nicotinamido ) Ethyl ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine 2- (1-methylpyridinium-3-amido) ethyl ester, N- (toluene-4-sulfonyl) -L-Prolyl-L-phenylalanine cholesteryl ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine 2- (1-methyl-1,4-dihydropyridinyl-3 -Amido) ethyl ester, N- (thiophen-2-sulfonyl) -L-prolyl-L-phenylalanine methyl ester, N- (thiophen-2-sulfonyl) -L-prolyl-L-phenylalanine , N- (5-chloro-1,3-dimethylpyrazole-4-sulfonyl) -L-prolyl-L-phenylalanine, N- (2-phenylethanesulfonyl) -L-prolyl-L-phenylalanine , N- (1-methylimidazole-4-sulfonyl) -L-prolyl-L-phenylalanine methyl ester, N- (1-methylimidazole-4-sulfonyl) -L-prolyl-L -Phenylalanine, N- (4-aminobenzenesulfonyl) -L-prop Aryl-L-phenylalanine methyl ester, N- (4-aminobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-thiomethoxyimidatylbenzene-4-sulfonyl) -L-prolyl -L-phenylalanine methyl ester, N- [4- (N-methylthioamido) benzenesulfonyl] -L-prolyl-L-phenylalanine methyl ester, N- (toluene-4-sulfonyl) -L-prop Rollyl-D, L-β- (1,2,4-triazol-3-yl) alanine, N- (toluene-4-sulfonyl) -L-proline-D, L-β- (thiazole-2 -Yl) alanine, N- [4- (3-dimethylaminopropoxy) benzenesulfonyl] -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-pyrrolidine-2 -Yl-thiocarbonyl-L-phenylalanine, N- (4-thiocarbamoylbenzenesulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester, N- (4-cyano Benzenesulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L-prolyl-D-phenylalanine, N- (toluene-4 -Sulfonyl) -L- (thiamorpholine-3-carbonyl) -L-phenylalanine, N- (toluene-4- Ponyl)-[(1,1-dioxo) thiamorpholine-3-carbonyl] -L-phenylalanine, N- (toluene-4-sulfonyl) -L- (3,3-dimethyl) prolyl-L -Phenylalanine, N- (toluene-4-sulfonyl) -L- (5,5-dimethyl-1,1-dioxo) thiaprolyl-L-phenylalanine, N- (toluene-4-sulfonyl)-[ (1,1-dioxo) thiamorpholine-3-carbonyl] -L-phenylalanine ethyl ester, N- (toluene-4-sulfonyl) -L-proline-L-phenylalanine t-butyl ester, N- ( Toluene-4-sulfonyl) -L-pyrrolidin-2-yl-thiocarbonyl-L-phenylalanine methyl ester, N- (benzenesulfonyl) -L-prolyl-L-phenylalanine methyl ester, N- ( Toluene-4-sulfonyl) -L- (5-oxo) prolyl-L-phenylalanine ethyl ester, N- (toluene-4-sulfonyl) -L- (5-oxo) prolyl-L-phenylalanine; And pharmaceutically acceptable salts thereof, as well as one ester consisting of methyl ester, ethyl ester, n-propyl ester, isopropyl ester, n-butyl ester, isobutyl ester, secondary butyl ester and tertiary butyl ester Any ester compound recited above, substituted with another ester selected from the group.

The present invention also provides a method of binding VLA-4 in a biological sample, including contacting the compound of Formula I or IA described above with a biological sample under conditions that the compound binds to VLA-4.

Certain compounds of Formula I and Formula IA above are also useful for reducing VLA-4 mediated inflammation in vivo.

The present invention also contains a therapeutically effective amount of at least one compound of Formula I or Formula IA and a pharmaceutically acceptable carrier, except that R ³ and R ⁵ are derived from L-amino acids or other similar forms of starting material. It relates to a pharmaceutical composition. Racemic compounds can also be used.

Pharmaceutical compositions can be used to treat VLA-4 mediated disease states. Such disease states include, for example, asthma, Alzheimer's disease, atherosclerosis, AIDS dementia, diabetes (including acute pediatric diabetes), inflammatory bowel disease (including ulcerative colitis and Crohn's disease), multiple sclerosis, rheumatoid arthritis, Acute leukocyte mediated lung injury, such as that occurs in tissue transplantation, tumor metastasis, meningitis, encephalitis, ataxia and other cerebral trauma, nephritis, retinitis, atopic dermatitis, psoriasis, myocardial ischemia, and adult respiratory distress syndrome.

Accordingly, the present invention also provides a method of treating a VLA-4 mediated inflammatory disease in a patient, comprising administering the pharmaceutical composition described above to the patient.

Preferred compounds of formula (I) and formula (IA) include those shown in Table I below.

As indicated above, the present invention relates to compounds which inhibit leukocyte adhesion, in particular leukocyte adhesion mediated by VLA-4. However, before further describing the present invention in detail, the following terms are first defined.

Justice

As used herein, "alkyl" is preferably an alkyl group having 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms. Such terms are exemplified by groups such as methyl, t-butyl, n-heptyl and octyl and the like.

"Substituted alkyl" refers to alkoxy, substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonyl Amino, aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl, cyano, halogen, hydroxyl, nitro, carboxyl, carboxylalkyl, carboxyl-substituted Alkyl, carboxy cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxyheteroaryl, carboxyl-substituted heteroaryl, carboxyheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl, substituted Cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl, substituted thiocycloalkyl, Thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heteroaryl, substituted aryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy, substituted cyclo Alkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -0S (O) ₂ -alkyl, -OS (O) ₂ -Substituted alkyl, -OS (O) ₂ -substituted alkyl, -OS (O) ₂ -aryl, -OS (O) ₂ -substituted aryl, -OS (O) ₂ -heteroaryl, -OS (O ) ₂ -substituted heteroaryl, -OS (O) ₂ -heterocyclic, -OS (O) ₂ -substituted heterocyclic, -OS (O) ₂ -NRR, wherein R is hydrogen or alkyl , -NRS (O) ₂ -alkyl, -NRS (O) ₂ -substituted alkyl, -NRS (O) ₂ -aryl, -NRS (O) ₂ -substituted aryl, -NRS (O) ₂ -heteroaryl , -NRS (O) ₂ -substituted heteroaryl, -NRS (O) ₂ -heterocyclic, -NRS ( O) ₂ -substituted heterocyclic, -NRS (O) ₂ -NR-alkyl, -NRS (O) ₂ -NR-substituted alkyl, -NRS (O) ₂ -NR-aryl, -NRS (O) ₂ -NR-substituted aryl, -NRS (O) ₂ -NR-heteroaryl, -NRS (O) ₂ -NR-substituted heteroaryl, -NRS (O) ₂ -NR-heterocyclic, -NRS ( O) ₂ -NR-substituted heterocyclic where R is hydrogen or alkyl, mono and dialkylamino, mono and di (substituted alkyl) amino, mono and diarylamino, disubstituted and disubstituted arylamino Mono and diheteroarylamino, mono and disubstituted heteroarylamino, mono and diheterocyclic amino, mono and disubstituted heterocyclic amino, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, Difference with asymmetric disubstituted amines having different substituents selected from substituted heteroaryls, heterocyclics and substituted heterocyclics, conventional blocking groups (eg Boc, Cbz, formyl, etc.) A substituted alkyl group, an amino group having a -SO ₂ -alkyl, -SO ₂ - substituted alkyl, -SO ₂ - alkenyl, -SO ₂ - substituted alkenyl, -SO ₂ cycloalkyl, -SO ₂ -Substituted cycloalkyl, -SO ₂ -aryl, -SO ₂ -substituted aryl, -SO ₂ -heteroaryl, -SO ₂ -substituted heteroaryl, -SO ₂ -heterocyclic, -SO ₂ -substituted Alkyl groups having 1 to 5 substituents selected from heterocyclic and alkyl / substituted alkyl groups substituted with —SO ₂ NRR, wherein R is hydrogen or alkyl, preferably from 1 to 10 carbon atoms.

"Alkoxy" is, for example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, t-butoxy, s-butoxy, n-pentoxy, n-hexoxy and Group "alkyl-O", including 1,2-dimethylbutoxy and the like.

"Substituted alkoxy" is a group "substituted alkyl-O-".

"Acyl" refers to the group HC (O)-, alkyl-C (O)-, substituted alkyl-C (O)-, alkenyl-C (O)-, substituted alkenyl-C (O)-, alky Neyl-C (O)-, substituted alkynyl-C (O)-, cycloalkyl-C (O)-, substituted cycloalkyl-C (O)-, aryl-C (O)-, substituted aryl -C (O)-, heteroaryl-C (O)-, substituted heteroaryl-C (O)-, heterocyclic-C (O)-and substituted heterocyclic-C (O)-, where , Alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted Heterocyclic is as defined above).

"Acylamino" refers to the group -C (O) NRR, wherein each R is independently hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted Is selected from the group consisting of aryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, each R in combination with a nitrogen atom to be heterocyclic or substituted hetero Cyclic rings where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, Heterocyclic and substituted heterocyclic as defined above).

"Thiocarbonylamino" refers to a group -C (S) NRR, wherein each R is independently hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted Aryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, each R is bonded to a nitrogen atom to form a heterocyclic or substituted Heterocyclic rings, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl , Heterocyclic and substituted heterocyclic are as defined above).

"Acyloxy" refers to the group alkyl-C (O) O-, substituted alkyl-C (O) O-, alkenyl-C (O) O-, substituted alkenyl-C (O) O-, alkynyl -C (O) O-, substituted alkynyl-C (O) O-, aryl-C (O) O-, substituted aryl-C (O) O-, cycloalkyl-C (O) O-, Substituted cycloalkyl-C (O) O-, heteroaryl-C (O) O-, substituted heteroaryl-C (O) O-, heterocyclic-C (O) O-, and substituted heterocyclic -C (O) O- (where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, heterocyclic and substituted heterocyclic are as defined above).

"Alkenyl" is an alkenyl group having one or more alkenyl unsaturated positions, preferably one or two, preferably 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms.

"Substituted alkenyl" refers to alkoxy, substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbon Nylamino, aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl, carboxyl substituted Alkyl, carboxyl cycloalkyl, carboxyl substituted cycloalkyl, carboxylaryl, carboxyl substituted aryl, carboxyheteroaryl, carboxyl substituted heteroaryl, carboxyl heterocyclic, carboxyl substituted heterocyclic, cycloalkyl, substituted cycloalkyl, Guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl, substituted thiocycloal , Thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy, substituted cycloalkoxy, hetero Aryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -0S (O) ₂ -alkyl, -OS (O) ₂ -substituted Alkyl, -OS (O) ₂ -aryl, -OS (O) ₂ -substituted aryl, -OS (O) ₂ -heteroaryl, -OS (O) ₂ -substituted heteroaryl, -OS (O) ₂ -Heterocyclic, -OS (O) ₂ -substituted heterocyclic, -OS (O) ₂ -NRR, wherein R is hydrogen or alkyl, -NRS (O) ₂ -alkyl, -NRS (O ) ₂ -substituted alkyl, -NRS (O) ₂ -aryl, -NRS (O) ₂ -substituted aryl, -NRS (O) ₂ -heteroaryl, -NRS (O) ₂ -substituted heteroaryl,- NRS (O) ₂ -heterocyclic, -NRS (O) ₂ -substituted heterocyclic, -NRS (O) ₂ -NR-alkyl, -NRS (O) ₂ -NR-substituted alkyl, -NRS (O) ₂ -NR-aryl, -NRS (O) ₂ -NR-substituted aryl, -NRS (O ) ₂ -NR-heteroaryl, -NRS (O) ₂ -NR-substituted heteroaryl, -NRS (O) ₂ -NR-heterocyclic, -NRS (O) ₂ -NR-substituted heterocyclic ( Wherein R is hydrogen or alkyl), mono and dialkylamino, mono and di (substituted alkyl) amino, mono and diarylamino, mono and disubstituted arylamino, mono and diheteroarylamino, monocyclic and Disubstituted heteroarylamino, mono and diheterocyclic amino, mono- and disubstituted heterocyclic amino, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted Asymmetric disubstituted amines with different substituents selected from heterocyclic, substituted with amino groups blocked with conventional blocking groups (e.g., Boc, Cbz, formyl, etc.) Alkenyl group or -SO ₂ -alkyl, -SO ₂ -substituted alkyl, -SO ₂ -alkenyl, -SO ₂ -substituted alkenyl, -SO ₂ -cycloalkyl, -SO ₂ -substituted cycloalkyl, -SO ₂ -aryl, -SO ₂ -substituted aryl, -SO ₂ -heteroaryl, -SO ₂ -substituted heteroaryl, -SO ₂ -heterocyclic, -SO ₂ -substituted heterocyclic and -SO An alkenyl group having 1 to 5 substituents selected from the group consisting of alkyl / substituted alkyl groups substituted with ₂ NRR, wherein R is hydrogen or alkyl.

"Alkynyl" is an alkynyl group having 2 to 10, more preferably 3 to 6, carbon atoms having one or more, preferably one or two, alkynyl unsaturated positions.

"Substituted alkynyl" refers to alkoxy, substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbon Nylamino, aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro, carboxyl, carboxylalkyl, carboxyl substituted Alkyl, carboxyl cycloalkyl, carboxyl substituted cycloalkyl, carboxylaryl, carboxyl substituted aryl, carboxyheteroaryl, carboxyl substituted heteroaryl, carboxyl heterocyclic, carboxyl substituted heterocyclic, cycloalkyl, substituted cycloalkyl, Guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl, substituted thiocycloal , Thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heteroaryl, substituted aryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy, substituted Cycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -0S (O) ₂ -alkyl, -OS (O) ₂ -substituted alkyl, -OS (O) ₂ -aryl, -OS (O) ₂ -substituted aryl, -OS (O) ₂ -heteroaryl, -OS (O) ₂ -substituted heteroaryl, -OS (O) ₂ -heterocyclic, -OS (O) ₂ -substituted heterocyclic, -OS (O) ₂ -NRR, wherein R is hydrogen or alkyl, -NRS (O) ₂ -alkyl, -NRS (O) ₂ -substituted alkyl, -NRS (O) ₂ -aryl, -NRS (O) ₂ -substituted aryl, -NRS (O) ₂ -heteroaryl, -NRS (O) ₂ -substituted Heteroaryl, -NRS (O) ₂ -heterocyclic, -NRS (O) ₂ -substituted hetero Cyclic, -NRS (O) ₂ -NR-alkyl, -NRS (O) ₂ -NR-substituted alkyl, -NRS (O) ₂ -NR-aryl, -NRS (O) ₂ -NR-substituted aryl , -NRS (O) ₂ -NR-heteroaryl, -NRS (O) ₂ -NR-substituted heteroaryl, -NRS (O) ₂ -NR-heterocyclic, -NRS (O) ₂ -NR-substituted Heterocyclic, wherein R is hydrogen or alkyl, mono and dialkylamino, mono and di (substituted alkyl) amino, mono and diarylamino, mono and disubstituted arylamino, mono and diheteroaryl Amino, mono- and di-substituted heteroarylamino, mono and diheterocyclic amino, mono- and di-substituted heterocyclic amino, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, hetero Asymmetric disubstituted amines having different substituents selected from cyclic and substituted heterocyclic, amino groups blocked with conventional blocking groups (e.g., Boc, Cbz, formyl, etc.) Substituted alkynyl group or -SO ₂ -alkyl, -SO ₂ -substituted alkyl, -SO ₂ -alkenyl, -SO ₂ -substituted alkenyl, -SO ₂ -cycloalkyl, -SO ₂ -substituted Cycloalkyl, -SO ₂ -aryl, -SO ₂ -substituted aryl, -SO ₂ -heteroaryl, -SO ₂ -substituted heteroaryl, -SO ₂ -heterocyclic, -SO ₂ -substituted heterocyclic And an alkynyl group having 1 to 5 substituents selected from the group consisting of alkynyl / substituted alkynyl groups substituted with —SO ₂ NRR where R is hydrogen or alkyl.

"Amidino" is a group And the term "alkylamidino" refers to one to three alkyl groups, such as ) Compound.

"Thioamidino" is a group Wherein R is hydrogen or alkyl.

"Aminoacyl" refers to the group -NRC (O) alkyl, -NRC (O) substituted alkyl, -NRC (O) cycloalkyl, -NRC (O) substituted cycloalkyl, -NRC (O) alkenyl, -NRC (O) substituted alkenyl, -NRC (O) alkynyl, -NRC (O) substituted alkynyl, -NRC (O) aryl, -NRC (O) substituted aryl, -NRC (O) heteroaryl, -NRC (O) substituted heteroaryl, -NRC (O) heterocyclic and -NRC (O) substituted heterocyclic, wherein R is hydrogen or alkyl and alkyl, substituted alkyl, alkenyl, substituted Alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic are as defined above).

"Aminocarbonyloxy" refers to the group -NRC (O) O-alkyl, -NRC (O) O-substituted alkyl, -NRC (O) O-alkenyl. -NRC (O) O-substituted alkenyl, -NRC (O) O-alkynyl, -NRC (O) O-substituted alkynyl, -NRC (O) O-cycloalkyl, -NRC (O) O -Substituted cycloalkyl, -NRC (O) O-aryl, -NRC (O) O-substituted aryl, -NRC (O) O-heteroaryl, -NRC (O) O-substituted heteroaryl, -NRC (O) O-heterocyclic and -NRC (O) O-substituted heterocyclic, wherein R is hydrogen or alkyl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted Alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic are as defined above).

"Oxycarbonylamino" refers to the group -OC (O) NH ₂ , -OC (O) NRR, -OC (O) NR-alkyl, -OC (O) NR-substituted alkyl, -OC (O) NR- Alkenyl, -OC (O) NR-substituted alkenyl, -OC (O) NR-alkynyl, -OC (O) NR-substituted alkynyl, -OC (O) NR-cycloalkyl, -OC ( O) NR-substituted cycloalkyl, -OC (O) NR-aryl, -OC (O) NR-substituted aryl, -OC (O) NR-heteroaryl, -OC (O) NR-substituted heteroaryl , -OC (O) NR-heterocyclic and -OC (O) NR-substituted heterocyclic, wherein R is hydrogen or alkyl, or each R is bonded to a nitrogen atom to be heterocyclic or substituted hetero Forms a cyclic ring, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl , Heterocyclic and substituted heterocyclic are as defined above).

"Oxythiocarbonylamino" refers to the groups -OC (S) NH ₂ , -OC (S) NRR, -OC (S) NR-alkyl, -OC (S) NR-substituted alkyl, OC (S) NR- Alkenyl, OC (S) NR-substituted alkenyl, OC (S) NR-alkynyl, OC (S) NR-substituted alkynyl, OC (S) NR-cycloalkyl, OC (S) NR-substituted Cycloalkyl, OC (S) NR-aryl, OC (S) NR-substituted aryl, OC (S) NR-heteroaryl, OC (S) NR-substituted heteroaryl, OC (S) NR-heterosai Click and OC (S) NR-substituted heterocyclic, wherein R is hydrogen or alkyl, or each R combines with a nitrogen atom to form a heterocyclic or substituted heterocyclic ring, alkyl, substituted Alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic As defined above).

"Aminocarbonylamino" refers to the group -NRC (O) NRR, -NRC (O) NR-alkyl, -NRC (O) NR-substituted alkyl, -NRC (O) NR-alkenyl, -NRC (O) NR-substituted alkenyl, -NRC (O) NR-alkynyl, -NRC (O) NR-substituted alkynyl, -NRC (O) NR-aryl, -NRC (O) NR-substituted aryl,- NRC (O) NR-cycloalkyl, -NRC (O) NR-substituted cycloalkyl, -NRC (O) NR-heteroaryl, -NRC (O) NR-substituted heteroaryl, -NRC (O) NR- Heterocyclic and -NRC (O) NR-substituted heterocyclic wherein R is independently of each other hydrogen or alkyl, or each R is bonded to a nitrogen atom to form a heterocyclic or substituted heterocyclic ring One of the amino groups is interrupted by conventional blocking groups (e.g., Boc, Cbz, formyl, etc.) and is substituted by alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cyclo Alkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle And substituted heterocyclic are as defined above].

"Aminothiocarbonylamino" refers to the group -NRC (S) NRR, -NRC (S) NR-alkyl, -NRC (S) NR-substituted alkyl, -NRC (S) NR-alkenyl, -NRC (S ) NR-substituted alkenyl, -NRC (S) NR-alkynyl, -NRC (S) NR-substituted alkynyl, -NRC (S) NR-aryl, -NRC (S) NR-substituted aryl, -NRC (S) NR-cycloalkyl, -NRC (S) NR-substituted cycloalkyl, -NRC (S) NR-heteroaryl, -NRC (S) NR-substituted heteroaryl, -NRC (S) NR -Heterocyclic, and -NRC (S) NR-substituted heterocyclic wherein R is independently of each other hydrogen or alkyl, or each R is bonded to a nitrogen atom to form a heterocyclic or substituted heterocyclic ring Wherein one of the amino groups is blocked by conventional blocking groups (e.g., Boc, Cbz, formyl, etc.), and are alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl , Cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, hetero yarn Cyclic and substituted heterocyclics are as defined above].

"Aryl" or "Ar" is an unsubstituted aromatic carbocyclic group having 6 to 14 carbon atoms having a single ring (eg phenyl) or multiple condensed rings (eg naphthyl or anthryl), wherein the condensed ring [Eg, 2-benzoxazolinone, 2H-1,4-benzoxazin-3 (4H) -one-7yl, etc.] may or may not be aromatic. Preferred aryls include phenyl and naphthyl.

"Substituted aryl" means hydroxy, acyl, acylamino, thiocarbonylamino, acryloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, Amidino, alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substituted Cycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, carboxyl, carboxylalkyl, carboxyl substituted alkyl, carboxyl cycloalkyl, carboxyl substituted cycloalkyl, carboxylaryl, carboxyl Substituted aryl, carboxyheteroaryl, carboxyl substituted heteroaryl, carboxyheterocyclic, carboxyl substituted heterocyclic, carboxyamido, cyano, thiol, thio Chel, substituted thioalkyl, thioaryl, substituted thioaryl, thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic, substituted thioheterocyclic, cycloalkyl, Substituted cycloalkyl, guanidino, guanidinosulfone, halo, nitro, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, oxycarbonylamino, oxythiocarbonylamino, -S ( O) ₂ -alkyl, -S (O) ₂ substituted alkyl, -S (O) ₂ -cycloalkyl, -S (O) ₂ -substituted cycloalkyl, -S (O) ₂ -alkenyl, -S (O) ₂ -substituted alkenyl, -S (O) ₂ -aryl, -S (O) ₂ -substituted aryl, -S (O) ₂ -heteroaryl, -S (O) ₂ -substituted hetero Aryl, -S (O) ₂ -heterocyclic, -S (O) ₂ -substituted heterocyclic, -S (O) ₂ -alkyl, -S (O) ₂ -substituted alkyl, -S (O ) _2-aryl, -S (O) aryl, -S (O ₂ substituted) _{2-heteroaryl,} -S (O) _{2-substituted} H. Loa reel, -S (O) ₂ - heterocyclic, -S (O) ₂ - substituted heterocyclic, -OSO ₂ NRR (wherein, R is hydrogen or alkyl), -NRS (O) ₂ - alkyl, , NRS (O) ₂ -substituted alkyl, NRS (O) ₂ -aryl, NRS (O) ₂ -substituted aryl, NRS (O) ₂ -heteroaryl, NRS (O) ₂ -substituted heteroaryl, NRS (O) ₂ - heterocyclic, NRS (O) ₂ - cyclic substituted heteroaryl, NRS (O) ₂ -NR- alkyl, NRS (O) ₂ -NR- substituted alkyl, NRS (O) ₂ -NR -Aryl, NRS (O) ₂ -NR-substituted aryl, NRS (O) ₂ -NR-heteroaryl, NRS (O) ₂ -NR-substituted heteroaryl, NRS (O) ₂ -NR-heterocyclic And NRS (O) ₂ -NR-substituted heterocyclic where R is hydrogen or alkyl, mono and dialkylamino, mono and di (substituted alkyl) amino, mono and diarylamino, monocyclic and Disubstituted heteroarylamino, mono and diheterocyclic amino, mono- and disubstituted heterocyclic amino, alkyl, substituted alkyl, aryl, substituted aryl, he In substituted aryls blocked with asymmetric disubstituted amines having different substituents selected from roaryl, substituted heteroaryls, heterocyclics and substituted heterocyclics and conventional blocking groups (e.g., Boc, Cbz, formyl, etc.) Or an aryl group substituted with 1 to 3 substituents selected from the group consisting of amino groups or substituted with —SO ₂ NRR where R is hydrogen or alkyl.

"Aryloxy" is a group aryl-O-, including, for example, phenoxy, naphthoxy, and the like.

"Substituted aryloxy" is a substituted aryl-O- group.

"Aryloxyaryl" is a group aryl-O-aryl.

"Substituted aryloxyaryl" means hydroxy, acyl, acylamino, thiocarbonylamino, acryloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substituted alkoxy Neyl, Amidino, Alkylamidino, Thioamidino, Amino, Aminoacyl, Aminocarbonyloxy, Aminocarbonylamino, Aminothiocarbonylamino, Aryl, Substituted aryl, Aryloxy, Substituted aryloxy, Cycloalkoxy , Substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl , Carboxylaryl, carboxyl-substituted aryl, carboxyheteroaryl, carboxyl substituted heteroaryl, carboxyl heterocyclic, carboxyl-substituted heterocyclic, carboxyamido, cyano, thi Ol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl, thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic, substituted thioheterocyclic, Cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, halo, nitro, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, oxycarbonylamino, oxythiocarbonylamino, -S (O) ₂ -alkyl, -S (O) ₂ -substituted alkyl, -S (O) ₂ -cycloalkyl, -S (O) ₂ -substituted cycloalkyl, -S (O) ₂ -al Kenyl, -S (O) ₂ -substituted alkenyl, -S (O) ₂ -aryl, -S (O) ₂ -substituted aryl, -S (O) ₂ -heteroaryl, -S (O) ₂ -Substituted heteroaryl, -S (O) ₂ -heterocyclic, -S (O) ₂ -substituted heterocyclic, -S (O) ₂ -alkyl, -S (O) ₂ -substituted alkyl, -S (O) _2-aryl, -S (O) _2-aryl, -S (O) _{2-substituted-heteroaryl,} -S (O) ₂ - Hwandoen heteroaryl, -S (O) ₂ - heterocyclic, -S (O) ₂ - cyclic substituted heteroaryl, -S (O) ₂ NRR (wherein, R is hydrogen or alkyl), -NRS (O ) ₂ -alkyl, -NRS (O) ₂ -substituted alkyl, -NRS (O) ₂ -aryl, -NRS (O) ₂ -substituted aryl, -NRS (O) ₂ -heteroaryl, -NRS (O ) ₂ -substituted heteroaryl, -NRS (O) ₂ -heterocyclic, -NRS (O) ₂ -substituted heterocyclic, -NRS (O) ₂ -NR-alkyl, -NRS (O) _2- NR- substituted alkyl, -NRS (O) ₂ -NR- aryl, -NRS (O) ₂ -NR- substituted aryl, -NRS (O) ₂ -NR- heteroaryl, -NRS (O) ₂ -NR -Substituted heteroaryl, -NRS (O) ₂ -NR-heterocyclic and -NRS (O) ₂ -NR-substituted heterocyclic, wherein R is hydrogen or alkyl, mono and dialkylamino, Mono and di (substituted alkyl) amino, mono and diarylamino, mono and disubstituted arylamino, mono and diheteroarylamino, mono and disubstituted heteroarylamino, mono and diheterocyclic amino, one Asymmetric disubstituted amines having different substituents selected from substituted and disubstituted heterocyclic amino, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic and conventional One or both aryl rings are substituted with one to three substituents selected from the group consisting of amino groups in substituted aryls blocked with phosphorus blocking groups (e.g., Boc, Cbz, formyl, etc.), or -SO ₂ NRR (where R Is hydrogen or alkyl).

"Cycloalkyl" is a cyclic alkyl group having 3 to 8 carbon atoms having a single cyclic ring including, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like. This definition excludes polycyclic alkyl groups such as adamantanyl and the like.

A "cycloalkenyl" is a cyclic alkenyl group having 3 to 8 carbon atoms that is single or multiple unsaturated but not aromatic.

"Substituted cycloalkyl" and "substituted cycloalkenyl" are oxo (= O), thioxo (= S), alkoxy, substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino, amino Dino, alkylamidino, thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxyaryl, substituted aryl Oxyaryl, halogen, hydroxy, cyano, nitro, carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxyheteroaryl, carboxyl -Substituted heteroaryl, carboxyheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulphone, thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted Done Thioaryl, thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heteroaryl, substituted heteroaryl, heterocyclic, substituted hetero Cyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -OS (O) ₂ -Alkyl, -OS (O) ₂ -substituted alkyl, -OS (O) ₂ -aryl, -OS (O) ₂ -substituted aryl, -OS (O) ₂ -heteroaryl, -OS (O) ₂ -Substituted heteroaryl, -OS (O) ₂ -heterocyclic, -OS (O) ₂ substituted heterocyclic, -OS (O) ₂ -NRR, wherein R is hydrogen or alkyl, -NRS (O) ₂ -alkyl, NRS (O) ₂ -substituted alkyl, NRS (O) ₂ -aryl, NRS (O) ₂ -substituted aryl, NRS (O) ₂ -heteroaryl, NRS (O) _2- substituted heteroaryl, NRS (O) ₂ - H. Rosayi click, NRS (O) ₂ - cyclic substituted heteroaryl, NRS (O) ₂ -NR- alkyl, NRS (O) ₂ -NR- substituted alkyl, NRS (O) ₂ -NR- aryl, NRS (O ) ₂ -NR-substituted aryl, NRS (O) ₂ -NR-heteroaryl, NRS (O) ₂ -NR-substituted heteroaryl, NRS (O) ₂ -NR-heterocyclic and NRS (O) ₂ -NR-substituted heterocyclic where R is hydrogen or alkyl, mono and dialkylamino, mono and di (substituted alkyl) amino, mono and diarylamino, mono and disubstituted heteroarylamino, Different mono and diheterocyclic amino, mono- and disubstituted heterocyclic amino, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic Substituted alkynyl groups having asymmetric disubstituted amines having substituents and amino groups blocked with conventional blocking groups (e.g., Boc, Cbz, formyl, etc.) and -SO ₂ -alkyl, -SO ₂ -substituted alkyl, -SO ₂ -alkenyl, -SO ₂ -substituted alkenyl, -SO ₂ -cycloalkyl, -SO ₂ -substituted cycloalkyl, -SO _2- Aryl, -SO ₂ -substituted aryl, -SO ₂ -heteroaryl, -SO ₂ -substituted heteroaryl, -SO ₂ -heterocyclic, -SO ₂ -substituted heterocyclic and -SO ₂ NRR where , R is hydrogen or alkyl), preferably a cycloalkyl group having 3 to 8 carbon atoms and a cycloalkenyl group having 1 to 5 substituents selected from alkynyl substituted alkynyl groups.

"Cycloalkoxy" is an -O-cycloalkyl group.

"Substituted cycloalkoxy" is an -O-substituted cycloalkyl group.

"Guanidino" is a group -NRC (= NR) NRR, -NRC (= NR) NR-alkyl, -NRC (= NR) NR-substituted alkyl, -NRC (= NR) NR-alkenyl, -NRC (= NR) NR-substituted alkenyl, -NRC (= NR) NR-alkynyl, -NRC (= NR) NR-substituted alkynyl, -NRC (= NR) NR-aryl, -NRC (= NR ) NR-substituted aryl, -NRC (= NR) NR-cycloalkyl, -NRC (= NR) NR-substituted cycloalkyl, -NRC (= NR) NR-heteroaryl, -NRC (= NR) NR- Substituted heteroaryl, -NRC (= NR) NR-heterocyclic and -NRC (= NR) NR-substituted heterocyclic, wherein each R is independently hydrogen and alkyl, and one of the amino groups is conventional Blocked with phosphorus blocking groups (e.g., Boc, Cbz, formyl, etc.) and substituted with alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, Substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic are as defined above].

"Guanidinosulfone" refers to a group -NRC (= NR) NRSO ₂ -alkyl, -NRC (= NR) NRSO ₂ -substituted alkyl, -NRC (= NR) NRSO ₂ -alkenyl, -NRC (= NR) NRSO ₂ -substituted alkenyl, -NRC (= NR) NRSO ₂ -alkynyl, -NRC (= NR) NRSO ₂ -substituted alkynyl, -NRC (= NR) NRSO ₂ -aryl, -NRC (= NR ) NRSO ₂ -substituted aryl, -NRC (= NR) NRSO ₂ -cycloalkyl, -NRC (= NR) NRSO ₂ -substituted cycloalkyl, -NRC (= NR) NRSO ₂ -heteroaryl, -NRC (= NR) NRSO ₂ -substituted heteroaryl, -NRC (= NR) NRSO ₂ -heterocyclic, and -NRC (= NR) NRSO ₂ -substituted heterocyclic, wherein each R is independently hydrogen and alkyl , Alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted Heterocyclic is as defined above].

"Halo" or "halogen" is fluorine, chlorine, bromine and iodine, preferably chlorine or bromine.

"Heteroaryl" is an aromatic carboxylic group having from 2 to 10 carbon atoms and having from 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur in the ring. Such heteroaryl groups may have a single ring (eg pyridyl or furyl) or multiple condensed rings (eg indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl, pyrrolyl, indolyl and furyl.

"Substituted heteroaryl" means hydroxy, acyl, acylamino, thiocarbonylamino, acryloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl , Amidino, alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, Substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, Carboxylaryl, carboxyl-substituted aryl, carboxyheteroaryl, carboxyl substituted heteroaryl, carboxyl heterocyclic, carboxyl-substituted heterocyclic, carboxyamido, cyano, thiol , Thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl, thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic, substituted thioheterocyclic, cyclo Alkyl, substituted cycloalkyl, guanidino, guanidinosulfone, halo, nitro, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, oxycarbonylamino, oxythiocarbonylamino,- S (O) ₂ -alkyl, -S (O) ₂ -substituted alkyl, -S (O) ₂ -cycloalkyl, -S (O) ₂ -substituted cycloalkyl, -S (O) ₂ -alkenyl , -S (O) ₂ -substituted alkenyl, -S (O) ₂ -aryl, -S (O) ₂ -substituted aryl, -S (O) ₂ -heteroaryl, -S (O) _2- Substituted heteroaryl, -S (O) ₂ -heterocyclic, -S (O) ₂ -substituted heterocyclic, -S (O) ₂ -alkyl, -S (O) ₂ -substituted alkyl,- S (O) ₂ - aryl, -S (O) ₂ - substituted aryl, -S (O) ₂ - heteroaryl, -S (O) ₂ - value Heteroaryl, -S (O) ₂ - heterocyclic, -S (O) ₂ - cyclic substituted heteroaryl, -OSO ₂ -NRR (wherein, R is hydrogen or alkyl), -NRS (O) ₂ -Alkyl, NRS (O) ₂ -substituted alkyl, NRS (O) ₂ -aryl, NRS (O) ₂ -substituted aryl, NRS (O) ₂ -heteroaryl, NRS (O) ₂ -substituted heteroaryl , NRS (O) ₂ -heterocyclic, NRS (O) ₂ -substituted heterocyclic, NRS (O) ₂ -NR-alkyl, NRS (O) ₂ -NR-substituted alkyl, NRS (O) ₂ -NR-aryl, NRS (O) ₂ -NR-substituted aryl, NRS (O) ₂ -NR-heteroaryl, NRS (O) ₂ -NR-substituted heteroaryl, NRS (O) ₂ -NR-hetero Cyclic and NRS (O) ₂ -NR-substituted heterocyclic where R is hydrogen or alkyl, mono and dialkylamino, mono and di (substituted alkyl) amino, mono and diarylamino, one Substituted and disubstituted heteroarylamino, mono and diheterocyclic amino, mono and disubstituted heterocyclic amino, alkyl, substituted alkyl, aryl, substituted Substituted asymmetric disubstituted amines having different substituents selected from reyl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic and blocked with conventional blocking groups (e.g., Boc, Cbz, formyl, etc.) Heteroaryl group substituted with 1 to 3 substituents selected from the group consisting of amino groups in aryl or substituted with —SO ₂ NRR where R is hydrogen or alkyl.

"Heteroaryloxy" is a group -O-heteroaryl and "substituted heteroaryloxy" is a group -O substituted heteroaryl.

A "heterocycle" or "heterocyclic" is a saturated that has from 1 to 10 carbon atoms, 1 to 4 heteroatoms selected from nitrogen, sulfur or oxygen in the ring, and at least one ring in the molten ring system may be aryl or heteroaryl. Or unsaturated groups.

A "saturated heterocyclic" is a heterocycle of a single or multiple condensed ring free of unsaturation in a ring (eg, carbon to carbon unsaturation, carbon to nitrogen unsaturation, nitrogen to nitrogen unsaturation, etc.).

An "unsaturated heterocyclic" is a non-aromatic heterocycle of a single or multiple condensed ring that is unsaturated in a ring (eg, carbon to carbon unsaturation, carbon to nitrogen unsaturation, nitrogen to nitrogen unsaturation, etc.).

"Substituted heterocyclic" means oxo (= O), thioxo (= S), alkoxy, substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino, thio Amidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl, halogen, hydroxy Cyano, nitro, carboxyl, carboxylalkyl, carboxyl substituted alkyl, carboxyl-cycloalkyl, carboxyl substituted cycloalkyl, carboxylaryl, carboxyl substituted aryl, carboxyheteroaryl, carboxyl substituted heteroaryl, carboxyheterocyclic, Carboxyl substituted heterocyclic, cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl, thiocyclo Chel, substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy , Substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -OS (O) ₂ -alkyl, -OS (O) ₂ -substituted alkyl, -OS (O) ₂ -aryl, -OS (O) ₂ -substituted aryl, -OS (O) ₂ -heteroaryl, -OS (O) ₂ -substituted heteroaryl , -OS (O) ₂ -heterocyclic, -OS (O) ₂ -substituted heterocyclic, -OSO ₂ -NRR, wherein R is hydrogen or alkyl, -NRS (O) ₂ -alkyl, NRS (O) ₂ -substituted alkyl, NRS (O) ₂ -aryl, NRS (O) ₂ -substituted aryl, NRS (O) ₂ -heteroaryl, NRS (O) ₂ -substituted heteroaryl, NRS ( O) ₂ - heterocyclic, NRS (O) ₂ - substituted Heterocyclic, NRS (O) ₂ -NR- alkyl, NRS (O) ₂ -NR- substituted alkyl, NRS (O) ₂ -NR- aryl, NRS (O) ₂ -NR- substituted aryl, NRS ( O) ₂ -NR-heteroaryl, NRS (O) ₂ -NR-substituted heteroaryl, NRS (O) ₂ -NR-heterocyclic and NRS (O) ₂ -NR-substituted heterocyclic, wherein R is hydrogen or alkyl), mono and dialkylamino, mono and di (substituted alkyl), mono and diarylamino, mono and disubstituted heteroarylamino, mono and diheterocyclic amino, mono and disubstituted Heterocyclic amino, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic asymmetric disubstituted amines having different substituents and conventional blocking groups Substituted alkynyl groups having amino groups blocked with (eg, Boc, Cbz, formyl, etc.) and -SO ₂ -alkyl, -SO ₂ -substituted alkyl, -SO ₂ -alkenyl, -SO ₂ -substituted alkenyl, -SO ₂ -cycloalkyl, -SO ₂ -substituted cycloalkyl, -SO ₂ -aryl, -SO ₂ -substituted aryl, -SO ₂ -heteroaryl , -SO ₂ - heteroaryl, -SO _{2-substituted-heterocyclic,} -SO ₂ - cyclic substituted heteroaryl, and -SO ₂ NRR with a carbonyl alkynyl / substituted (herein, R is hydrogen or alkyl) Heterocyclic group having 1 to 3 substituents selected from the group consisting of alkynyl groups.

Examples of heterocycles and heteroaryls include pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolysin, isotulin, Quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cynoline, phthalidine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole , Phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indolin, phthalimide, 1,2,3,4-tetraisoquinoline, 4,5,6,7 Tetrahydrobenzo [b] thiophene, thiazole, thiazolidine, thiophene, benzo [b] thiophene, morpholino, thiomorpholino, piperidinyl, pyrrolidine, tetrahydrofuranyl and the like. Including but not limited to.

A "saturated substituted heterocyclic" is a substituted heterocycle of a single or multiple condensed ring free of unsaturation in a ring (eg, carbon to carbon unsaturation, carbon to nitrogen unsaturation, nitrogen to nitrogen unsaturation, etc.).

An "unsaturated substituted heterocyclic" is a non-aromatic substituted heterocycle of a single or multiple condensed ring that is free of unsaturation in a ring (eg, carbon to probolic unsaturation, carbon to nitrogen unsaturation, nitrogen to nitrogen unsaturation, etc.).

"Heterocyclyloxy" is a group -O-heterocyclic, and "substituted heterocyclicoxy" is a group -O-substituted heterocyclic.

"Thiol" is the group -SH.

"Thioalkyl" is a group -S-alkyl.

"Substituted thioalkyl" is a group -S-substituted alkyl.

"Thiocycloalkyl" is a group -S-cycloalkyl.

"Substituted thiocycloalkyl" is a group -S-substituted cycloalkyl.

"Thioaryl" is a group -S-aryl and "substituted thioaryl" is a group -S-substituted aryl.

"Thioheteroaryl" is a group -S-heteroaryl and "substituted thioheteroaryl" is a group -S-substituted heteroaryl.

"Thioheterocyclic" is a group -S-heterocyclic, and "substituted thioheterocyclic" is a group -S-substituted heterocyclic.

"Pharmaceutically acceptable salts" are pharmaceutically acceptable salts of compounds of formula (I) or formula (IA) derived from various organic and inorganic counter ions, for example, only sodium, potassium, calcium, magnesium, ammonium, Tetraalkylammonium and the like, and when the compound of formula (I) or formula (IA) contains an organic functional group, a salt of an organic or inorganic acid (e.g., hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate And the like).

Compound manufacture

Compounds of the present invention can be prepared from commercially available starting materials using the following general methods and processes. It will be appreciated that other process conditions may be used, unless otherwise specified, given the usual or desired process conditions (ie reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.). Optimum reaction conditions may vary depending on the particular reactants or solvent used, but such conditions can be determined by those skilled in the art according to routine optimization procedures.

In addition, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent undesirable reactions from occurring in certain functional groups. Suitable protecting groups for various functional groups and suitable conditions for protecting and deprotecting certain functional groups are well known in the art. For example, many protection groups are described in T. W. Greene and G. M. Wuts, Proctecting Groups in Organic Synthesis, Second Edition, Wiley, New York, 1991, which is incorporated herein by reference.

In addition, the compounds of the present invention typically comprise one or more chiral centers. Thus, if desired, such compounds may be prepared or separated as pure stereoisomers (ie, each enantiomer or diastereomer) or as a mixture rich in stereoisomers. All such stereoisomers (and mixtures rich in them) are included within the scope of the present invention unless otherwise noted. Pure stereoisomers (or mixtures rich in them) can be prepared, for example, using optically active starting materials or stereoselective reagents well known in the art. In addition, racemic mixtures of such compounds can be separated using, for example, chiral column chromatography, chiral decomposition reagents and the like.

In a preferred method of synthesis, compounds of formula (I) and formula (IA) are prepared by first coupling an amino acid of formula (II) with a sulfonyl chloride of formula (III) to provide an N-sulfonyl amino acid of formula (IV).

R ¹ -SO ₂ -Cl

In the above formula (II), (III) and (IV),

R ¹ , R ² and R ³ are as defined in formula (I),

R ⁴ is hydrogen.

Such reactions are typically carried out by reacting at least one equivalent, preferably about 1.1 to about 2 equivalents of sulfonyl chloride of formula III in an inert diluent such as dichloromethane and the like. Generally, the reaction is carried out for about 1 to about 24 hours in the temperature range of about -70 to about 40 ℃. Preferably, this reaction is carried out in the presence of a suitable base to remove the acid produced during the reaction. Suitable bases include, for example, tertiary amines such as triethylamine, diisopropylethylamine, N-methylmorpholine and the like. The reaction can also be carried out under Schotten-Baumann-type conditions using an aqueous alkali (eg sodium hydroxide, etc.) as the base. Upon completion of the reaction, the resulting N-sulfonyl amino acid of formula IV is recovered by conventional methods including neutralization, extraction, precipitation, chromatography, filtration and the like.

The amino acid of formula (II) used in the above reaction is a known compound from a known compound or a compound prepared by a conventional synthetic process. Examples of amino acids suitable for use in this reaction include L-proline, trans-4-hydroxy-L-proline, cis-4-hydroxy-L-proline, trans-3-phenyl-L-proline, cis-3 -Phenyl-L-proline, L- (2-methyl) proline, L-pipecolic acid, L-indolin-2-carboxylic acid, L-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, L-thiazolidine-4-carboxylic acid, L- (5,5-dimethyl) thiazolidine-4-carboxylic acid, L-thiamorpholine-3-carboxylic acid, glycine, 2-t-butylglycine, D, L- Phenylglycine, L-alanine, α-methylalanine, N-methyl-L-phenylalanine, L-diphenylalanine, sacosine, D, L-phenylsacocine, L-aspartic acid β-t-butyl ester, L-glutamic acid γ-t-butyl ester, L- (O) -benzyl) serine, 1-aminocyclopropanecarboxylic acid, 1-aminocyclobutanecarboxylic acid, 1-aminocyclopentanecarboxylic acid (cycloleucine) 1-aminocyclohexanecarboxylic acid, L -Including but not limited to serine No. If desired, corresponding carboxylic esters of amino acids of the formula (II) such as methyl esters, ethyl esters and the like can be used in the above reaction with the sulfonyl chlorides of the formula (III). Conventional reagents and conditions are used to subsequently hydrolyze the ester groups with carboxylic acids, ie, with alkali metal hydroxides in an inert diluent such as methanol / water, to yield the N-sulfonyl amino acid of formula IV.

Similarly, the sulfonyl chloride of formula III used in the above reaction is a known compound or a compound which can be prepared from known compounds by conventional synthetic procedures. Such compounds are typically prepared from the corresponding sulfonic acids using phosphorus trichloride and phosphorus pentachloride, ie compounds of the formula R ¹ -SO ₃ H, wherein R ¹ is as defined above. Generally, this reaction is brought about by contacting the sulfonic acid with about 2 to about 5 molar equivalents of phosphorus trichloride and phosphorus pentachloride for about 1 to about 48 hours at about 0 to about 80 ° C. in a pure or inert solvent such as dichloromethane. Sulfonyl chloride is obtained. In addition, by treating thiols with chlorine (Cl ₂ ) and water under conventional reaction conditions of formula III, the corresponding thiol compounds, ie, the formula R ¹ -SH, wherein R ¹ are as defined above, can be prepared.

Examples of sulfonyl chlorides suitable for use in the present invention include methanesulfonyl chloride, 2-propanesulfonyl chloride, 1-butanesulfonyl chloride, benzenesulfonyl chloride, 1-naphthalenesulfonyl chloride, 2-naphthalenesulfonyl Chloride, p-toluenesulfonyl chloride, α-toluenesulfonyl chloride, 4-acetamidobenzenesulfonyl chloride, 4-amidinobenzenesulfonyl chloride, 4-t-butylbenzenesulfonyl chloride, 4-bromobenzene Sulfonyl chloride, 2-carboxybenzenesulfonyl chloride, 4-cyanobenzenesulfonyl chloride, 3,4-dichlorobenzenesulfonyl chloride, 3,5-dichlorobenzenesulfonyl chloride, 3,4-dimethoxybenzenesulfonyl Chloride, 3,5-ditrifluorobenzenesulfonyl chloride, 4-fluorobenzenesulfonyl chloride, 4-methoxybenzenesulfonyl chloride, 2-methoxycarbonylbenzenesulfonyl chloride, 4 -Methylamidobenzenesulfonyl chloride, 4-nitrobenzenesulfonyl chloride, 4-thioamidobenzenesulfonyl chloride, 4-trifluoromethylbenzenesulfonyl chloride, 4-trifluoromethoxybenzenesulfonyl chloride, 2 , 4,6-trimethylbenzenesulfonyl chloride, 2-phenylethanesulfonyl chloride, 2-thiophensulfonyl chloride, 5-chloro-2-thiophensulfonyl chloride, 2,5-dichloro-4-thiophensul Ponyl chloride, 2-thiazolesulfonyl chloride, 2-methyl-4-thiazolesulfonyl chloride, 1-methyl-4-imidazolesulfonyl chloride, 1-methyl-4-pyrazolesulfonyl chloride, 5-chloro-1, 3-dimethyl-4-pyrazolesulfonyl chloride, 3-pyridinesulfonyl chloride, 2-pyrimidinesulfonyl chloride, and the like. If desired, sulfonyl fluoride, sulfonyl bromide or sulfonic anhydride may be used in place of sulfonyl chloride in the above reaction to form the N-sulfonyl amino acid of formula IV.

The compound of formula I is then prepared by coupling the intermediate N-sulfonyl amino acid of formula IV with an amino acid derivative of formula VI.

In Formula VI above,

R ⁵ to R ⁷ are as defined in formula (I) and formula (IV).

This coupling reaction is carried out using typically well known coupling reagents such as carbodiimide, BOP reagent (benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphonate) and the like. . Suitable carbodiimides include, for example, dicyclohexylcarbodiimide (DCC), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (EDC), and the like. If desired, the polymer supported form of the carbodiimide coupling reagent may be used, including, for example, the compounds described in Tetrahedron Letters, 34 (48), 7685 (1993). In addition, well-known coupling promoters (eg, N-hydroxysuccinimide, 1-hydroxybenzotriazole, etc.) can be used to promote the coupling reaction.

Such coupling reactions typically comprise from about 1 to about 2 equivalents of N-sulfonylamino acid in an inert diluent (e.g., dichloromethane, chloroform, acetonitrile, tetrahydrofuran, N, N-dimethylformamide, etc.). And at least one equivalent of formula VI, preferably about 1 to about 1.2 equivalents. Generally, this reaction is carried out for about 12 to about 24 hours in the temperature range of about 0 to about 37 ° C. Upon completion of the reaction, the compound of formula I is recovered by conventional methods including neutralization, extraction, precipitation, chromatography, filtration and the like.

The N-sulfonyl amino acid of formula IV can also be converted to an acid halide coupled with an acid halide and an amino acid derivative of formula VI to yield a compound of formula I. Acid halides of formula VI can be prepared by contacting a compound of formula VI with an inorganic acid halide (eg, thionyl chloride, phosphorus trichloride, phosphorus tribromide or phosphorus pentachloride) or preferably oxalyl chloride under conventional conditions. Generally, this reaction is carried out in a pure or inert solvent such as dichloromethane or carbon tetrachloride using about 1 to 5 molar equivalents of inorganic acid halide or oxalyl chloride for about 1 to about 48 hours in the temperature range of about 0 to about 80 ° C. To perform. Catalysts such as N, N-dimethylformamide may also be used for this reaction.

The acid halide of N-sulfonyl amino acid (IV) of Formula IV is then dissolved in an inert diluent such as dichloromethane for about 1 to about 24 hours in a temperature range of about -70 to about -40 ° C. At least about 1 equivalent, preferably from about 1.1 to about 1.5 equivalents, of the amino acid derivative. Preferably, this reaction is carried out in the presence of a suitable base which eliminates the acid produced during the reaction. Suitable bases include, for example, tertiary amines such as triethylamine, diisopropylethylamine, N-methylmorpholine and the like. The reaction can also be carried out under Shoten-Baumann type conditions using an aqueous alkali (eg sodium hydroxide, etc.). Upon completion of the reaction, the compound of formula I is recovered by conventional methods including neutralization, extraction, precipitation, chromatography, filtration and the like.

In addition, a compound of formula (I) may be prepared by first forming a diamino acid derivative of formula (VII).

In Formula VII above,

R ² to R ⁷ are as defined above.

Diamino acid derivatives of formula (VII) can be readily prepared by coupling amino acids of formula (II) with amino acid derivatives of formula (VI) using conventional amino acid coupling techniques and reagents such as carbodiimide, BOP reagents, and the like. The diamino acid of formula (VII) can then be sulfonated using the sulfonyl chloride of formula (III) and the synthesis described above to obtain a compound of formula (I).

The amino acid derivatives of formula VI used in the above reactions are known compounds or compounds which can be prepared from known compounds by conventional synthetic procedures. For example, amino acid derivatives of Formula VI can be prepared by C-alkylating commercially available diethyl 2-acetamidomalonate (Alrich, Milwaukee, WI) with alkyl or substituted alkyl halides. . This reaction is commonly carried out by treating diethyl 2-acetamidomalonate with at least one equivalent of sodium ethoxide and at least one equivalent of alkyl or substituted alkyl halide in reflux ethanol for about 6 to about 12 hours. The resulting C-alkylated malonate is then deacetylated and then hydrolyzed and decarboxylated by heating in aqueous hydrochloric acid under reflux for about 6 to about 12 hours to afford amino acids, typically as hydrochloric acid salts.

Examples of amino acid derivatives of formula VI suitable for use in the above reactions include L-tryptophan methyl ester, L-phenylalanine methyl ester, L-phenylalanine isopropyl ester, L-phenylalanine benzyl ester, L-phenylalanineamide, N-methyl- L-phenylalanine benzyl ester, D, L-homophenylalanine methyl ester, β- (1-naphthyl) -L-alanine methyl ester, β- (2-naphthyl) -L-alanine methyl ester, β- (2- Thienyl) -L-alanine methyl ester, β- (2-pyridyl) -L-alanine methyl ester, β- (3-pyridyl) -L-alanine methyl ester, β- (4-pyridyl) -L -Alanine methyl ester, β- (2-thiazolyl) -D, L-alanine methyl ester, β- (1,2,4-thiazol-3-yl) -D, L-alanine methyl ester, and the like. However, the present invention is not limited thereto. Of course, if desired, esters or amides other than the compounds described above may be used.

For ease of synthesis, compounds of formula (I) are typically prepared as esters, i.e., as alkoxy or substituted alkoxy groups in which R ⁶ is substituted. If desired, ester groups can be hydrolyzed using conventional conditions and reagents to provide the corresponding carboxylic acids. Typically, this reaction involves esters in an inert diluent (e.g. methanol or a mixture of methanol and water) for at least one equivalent of alkali metal hydroxide (e.g. lithium hydroxide) for about 1 to about 12 hours in a temperature range of about 0 to about 24 ° C. , Sodium hydroxide or potassium hydroxide). Benzyl esters can also be removed by hydrolysis using a palladium catalyst (eg palladium on carbon). If desired, the resulting carboxylic acid may be prepared using the conventional coupling reagents and conditions described above, using amines such as β-alanine ethyl ester, hydroxyamines such as hydroxyamine, N-hydroxysuccinimide. ) Alkoxyamine and substituted alkoxyamines such as O-methylhydroxylamine, O-benzylhydroxylamine, and the like.

As will be apparent to those skilled in the art, other functional groups present in the substituents of compounds of formula I may be readily modified or derivatized before or after the coupling reactions described above using well known synthetic procedures. Can be. For example, the nitro group present in the intermediate of the substituent of the compound of formula (I) can be readily reduced by hydrogenation in the presence of a palladium catalyst (eg palladium on carbon) to give the corresponding amino group. This reaction is typically carried out in an inert diluent (eg methanol) at a temperature of about 20 to about 50 ° C. for about 6 to about 24 hours. Compounds having a nitro group in the R ⁵ substituent may be prepared, for example, using 4-nitrophenylalanine derivatives or the like in the coupling reaction described above.

Similarly, the pyridyl group can be hydrogenated in an acidic diluent in the presence of a platinum catalyst (eg platinum oxide) to provide the corresponding piperidinyl homologue. Generally, this reaction is carried out in an acidic diluent (eg, a mixture of methanol and aqueous hydrochloric acid) in the presence of a catalyst at a temperature of about 20 to about 50 ° C. at a pressure range of about 20 to about 60 psi, preferably about 40 psi. The pyridine compound is treated with hydrogen for from about 24 hours. Compounds having pyridyl groups can be used in the coupling reactions described above, for example, β- (2-pyridyl)-, β- (3-pyridyl)-or β- (4-pyridyl) -L- It can be easily prepared using an alanine derivative.

According to the description, compounds of formula (I) having a substituent comprising a primary or secondary amino group (R ¹ is a 4-aminophenyl group) or intermediates thereof are easily N-using conventional acylation reagents and conditions. Acylation affords the corresponding amide. This acylation reaction is carried out in an inert diluent in the presence of a coupling agent (e.g. carbodiimide, a BOP agent such as benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphonate, etc.). (Eg, dichloromethane, chloroform, acetonitrile, tetrahydrofuran, N, N-dimethylformamide, etc.) at least one equivalent, preferably about one amino compound, at about 0 to about 37 ° C. for about 4 to about 24 hours. Typically carried out by reaction with 1.1 to about 1.2 equivalents of carboxylic acid. Preferably, accelerators (eg, N-hydroxysuccinimide, 1-hydroxybenzotriazole, etc.) are used to promote the acylation reaction. Examples of suitable carboxylic acids for use in this reaction include Nt-butyloxycarbonylglycine, Nt-butyloxycarbonyl-L-phenylalanine, Nt-butyloxycarbonyl-L-aspartic acid benzyl ester, benzoic acid, Nt-butyloxy Carbonylisonifeconic acid, N-methylisonifeconic acid, Nt-butyloxycarbonylnifeconic acid, Nt-butyloxycarbonyl-L-tetrahydroisoquinoline-3-carboxylic acid, N- (toluene-4-sulfur Phonyl) -L-proline, and the like.

In addition, compounds of formula (I) or intermediates thereof comprising primary or secondary amino groups are N-acylated with acyl halides or carboxylic anhydrides to form the corresponding amides. This reaction comprises at least about 1 equivalent of acyl halides or carboxylic anhydrides, preferably from about 1.1 to about 1 to about 24 hours, of the amino compound in an inert solvent such as dichloromethane at a temperature ranging from about -70 to about 40 ° C. Typically performed in contact with about 1.2 equivalents. If desired, an acylation catalyst such as 4- (N, N-dimethylamino) pyridine can be used to promote the acyl reaction. The acylation reaction is preferably carried out in the presence of a suitable base to capture the acid produced during the reaction. Suitable bases include, for example, tertiary amines (eg triethylamine, diisopropylethylamine, N-methylmorpholine, etc.). The reaction can also be carried out under Shoten-Baumann type conditions using an aqueous alkali (eg sodium hydroxide, etc.).

Examples of acyl halides and carboxylic anhydrides suitable for use in this reaction include 2-methylpropionyl chloride, trimethylacetyl chloride, phenylacetyl chloride, benzoyl chloride, 2-bromobenzoyl chloride, 2-methylbenzoyl chloride, 2-trifluor Romethylbenzoyl chloride, isnicotinoyl chloride, nicotinoyl chloride, picolinoyl chloride, acetic anhydride, succinic anhydride, and the like. Carbamyl chloride, such as N, N-dimethylcarbamyl chloride, N, N-diethylcarbamyl chloride, may be used in this reaction to provide urea. Similarly, bicarbonates such as di-t-butyl dicarbonate can be used to provide carbonates.

In a similar manner, compounds of formula (I) or intermediates thereof comprising primary or secondary amino groups are N-sulfonated using sulfonyl halides or sulfonic anhydrides to form sulfonamides. Sulfonyl halides and sulfonic anhydrides suitable for this reaction include, but are not limited to, methanesulfonyl chloride, chloromethanesulfonyl chloride, p-toluenesulfonyl chloride, trifluoromethanesulfonic anhydride, and the like. Similarly, sulfamoyl chlorides such as dimethylsulfamoyl chloride may be used to provide sulfamides (eg,> N-SO ₂ -N <).

In addition, primary or secondary amino groups present in the substituents of compounds of formula (I) or intermediates thereof are reacted with isocyanates or thioisocyanates to obtain ureas or thioureas, respectively. Such reactions typically comprise at least one equivalent, preferably from about 1.1 to about 1.2 equivalents of isocyanate, in an inert diluent (e.g., toluene, etc.) for about 12 to about 24 hours in a temperature range of about 24 to about 37 ° C. Or by contacting with thioisocyanate. Isocyanates and thioisocyanates used in this reaction are either commercially available or can be prepared from commercially available compounds using well known synthetic procedures. For example, isocyanates and thioisocyanates are readily prepared by reacting suitable amines with phosgene or thiophosgene. Examples of suitable isocyanates and thioisocyanates for use in the reaction include ethyl isocyanates, n-propyl isocyanates, 4-cyanophenyl isocyanates, 3-methoxyphenyl isocyanates, 2-phenylethyl isocyanates, methyl thioisocyanates, ethyl thioisocyanates, 2-phenylethyl thioisocyanate, 3-phenylpropyl thioisocyanate, 3- (N, N-diethylamino) propyl thioisocyanate, phenyl thioisocyanate, benzyl thioisocyanate, 3-pyridyl thioisocyanate, fluorescein isothiocia Nate (isomer I) and the like, but are not limited thereto.

In addition, when the compound of formula (I) or an intermediate thereof comprises a primary or secondary amino group, the amino group is reductively alkylated with an aldehyde or ketone to form a secondary or tertiary amino group. Such reactions typically result in at least one equivalent of an aldehyde or ketone for about 1 to about 72 hours in an inert diluent (e.g., methanol, tetrahydrofuran, mixtures thereof, etc.) at a temperature ranging from about 0 to about 50 ° C, It is preferably carried out in contact with about 1.1 to about 1.5 equivalents and at least one equivalent of an amino compound-based metal hydroxide reducing agent such as sodium cyanoborohydride. Suitable aldehydes and ketones for use in the reaction include, for example, benzaldehyde, 4-chlorobenzaldehyde, valeraldehyde and the like.

In a similar manner, when the compound of formula (I) or intermediate thereof has a substituent comprising a hydroxyl group, the hydroxyl group is further modified or derivatized before or after the above coupling reaction, for example ether, Carbamate and the like.

For example, a compound of formula I having a substituent comprising a hydroxyl group, wherein R ¹ is a 4-hydroxyphenyl group, or a derivative thereof, is readily O-alkylated to form an ether. This O-alkylation reaction typically involves contacting a hydroxy compound with a suitable alkali or alkaline earth metal base (e.g. potassium carbonate) in an inert diluent (e.g. acetone, 2-butanone, etc.) to an alkali or alkaline earth metal salt of the hydroxyl group. It is carried out by forming a. Such salts are generally not separated, but are reacted with at least one equivalent of alkyl or substituted alkyl halides or sulfonates (eg alkyl chloride, bromide, iodide, mesylate or tosylate) in the reaction system to produce ethers. Generally, the reaction is carried out for about 24 to about 72 hours in the temperature range of about 60 to about 150 ° C. Preferably, when alkyl chloride or alkyl bromide is used in the reaction, a catalytic amount of sodium iodide or potassium iodide is added to the reaction mixture.

Examples of suitable alkyl or substituted alkyl halides and sulfonates for use in this reaction include t-butyl bromoacetate, Nt-butyl chloroacetamide, 1-bromoethylbenzene, ethyl α-bromophenylacetate, 2- (N-ethyl-N-phenylamino) ethyl chloride, 2- (N, N-ethylamino) ethyl chlorolide, 2- (N, N-diisopropylamino) ethyl chloride, 2- (N, N-di Benzylamino) ethyl chloride, 3- (N, N-ethylamino) propyl chloride, 3- (N-benzyl-N-methylamino) propyl chloride, N- (2-chloroethyl) morpholine, 2- (hexamethylene Imino) ethyl chloride, 3- (N-methylpiperazine) propyl chloride, 1- (3-chlorophenyl) -4- (3-chloropropyl) piperazine, 2- (4-hydroxy-4-phenylpi Ferridine) ethyl chloride, Nt-butyloxycarbonyl-3-piperidinemethyltosylate, and the like.

In addition, the hydroxyl groups present in the substituents of compounds of formula (I) or intermediates thereof may be O-alkylated using Mitsunobu reactions. In such a reaction, an alcohol such as 3- (N, N-dimethylamino) -1-propanol is in the inert diluent (e.g. tetrahydrofuran) in a temperature range of about -10 to about 5 ° C for about 0.25 to about 1 hour. For about 1.0 to about 1.3 equivalents of triphenylphosphine and about 1.0 to about 1.3 equivalents of diethyl azodicarboxylate. Then, about 1.0 to about 1.3 equivalents of hydroxy compound are added and the reaction mixture is stirred at a temperature of about 0 to about 30 ° C. for about 2 to about 48 hours to provide an O-alkylated product.

In a similar manner, a compound of formula (I) or an intermediate thereof containing an aryl hydroxy group is reacted with aryl iodide to give a diaryl ether. In general, the reaction is carried out in an inert diluent (eg xylene) at a temperature of about −25 to about 10 ° C. to form an alkali metal salt of the hydroxyl group with a suitable base (eg sodium hydroxide). Thereafter, the salt is treated with about 1.1 to about 1.5 equivalents of copper bromide dimethyl sulfide complex for about 0.5 to about 2.0 hours in the temperature range of about 10 to about 30 ° C, and then about 1.1 to about 1.5 equivalents of aryl iodide (E.g. sodium 2-iodobenzoate, etc.). Thereafter, the reaction is heated to a temperature of about 70 to about 150 ° C. for about 2 to about 24 hours to provide a diaryl ether.

In addition, the hydroxy containing compound may be readily derivatized to form carbamate. One of the methods for preparing such carbamate, is about 1.0 to about 1.2 of a hydroxy compound of formula (I) or an intermediate thereof in an inert diluent (e.g. dichloromethane) for about 0.5 to about 2.0 hours at about -25 to about 0 ° C. Contact with an equivalent of 4-nitrophenyl chloroformate. The resulting carbonate is treated with excess, preferably about 2 to about 5, equivalents of trialkylamine (e.g. triethylamine) for about 0.5 to about 2 hours, then about 1.0 to about 1.5 equivalents of primary or 2 Treatment with primary amines gives carbamate. Examples of suitable amines for use in the reaction include, but are not limited to, piperazine, 1-methylpiperazine, 1-acetylpiperazine, morpholine, thiomorpholine, pyrrolidine, piperidine, and the like.

In addition, as another method of preparing carbamate, the hydroxy containing compound may be added from about 1.0 to about 1.5 for about 2 to about 72 hours in an inert diluent (e.g., dichloromethane) at a temperature ranging from about 25 to about 70 ° C. Contact with an equivalent of carbamyl chloride. Typically, this reaction is carried out in the presence of a suitable base that captures the acid produced during the reaction. Suitable bases include, for example, tertiary amines such as triethylamine, diisopropylethylamine, N-methylmorpholine and the like. It is also preferred to add at least one equivalent of 4- (N, N-dimethylamino) pyridine (based on the hydroxy compound) to the reaction mixture to facilitate the reaction. Examples of carbamyl chloride suitable for use in the reaction include dimethylcarbamyl chloride, diethylcarbamyl chloride and the like.

In addition, when a compound of formula (I) or an intermediate thereof comprises primary or secondary hydroxyl groups, these hydroxyl groups are readily converted and replaced by leaving groups, for example to form amines, sulfides and fluorides. do. For example, derivatives of 4-hydroxy-L-proline can be converted to the corresponding 4-amino, 4-thio or 4-fluoro-L-proline derivatives via nucleophilic substitution of the derivatized hydroxyl groups. have. In general, when chiral compounds are used in these reactions, the stereochemistry at the carbon atom which is bound to the derivatized hydroxyl group is typically reversed.

These reactions are typically carried out by first converting the hydroxyl group to a leaving group (e.g. tosylate) by first treating the hydroxyl compound in pyridine with at least one equivalent of sulfonyl halide (e.g. p-toluenesulfonyl chloride, etc.). . This reaction is generally performed at a temperature of about 0 to about 70 ° C. for about 1 to about 48 hours. The resulting tosylate can then be subjected to tosylate for about 1 to about 12 hours, for example, in an inert diluent (eg, a mixture of N, N-dimethylformamide and water) at a temperature range of about 0 to about 37 ° C. It can be easily substituted by contact with at least one equivalent of sodium azide to provide the corresponding azide compound. The azido group is then hydrogenated using a palladium on carbon catalyst to reduce, for example, to provide an amino (-NH ₂ ) compound.

Similarly, tosylate groups are easily substituted with thiols to form sulfides. The reaction is typically carried out in an inert diluent (eg N, N-dimethylformamide) in the presence of a suitable base (eg 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU)). Tosylate is contacted with at least one equivalent of thiol (eg, thiophenol) at a temperature of 0 to about 37 ° C. for about 1 to about 12 hours to provide sulfides. Furthermore, tosylate is treated with morpholinosulfur trifluoride for about 12 to about 24 hours in a temperature range of about 0 to about 37 ° C. in an inert diluent such as dichloromethane to give the corresponding fluoro compound. .

In addition, a compound of formula (I) having a substituent comprising an iodoaryl group, wherein R ¹ is a 4-iodophenyl group, or an intermediate thereof, may be readily converted to a biaryl compound before or after the above coupling reaction. Can be. Typically, the reaction reacts the iodoaryl compound in an inert diluent (e.g. tetrahydrofuran) in the presence of a palladium catalyst (e.g. palladium tetra (triphenylphosphine)) at a temperature ranging from about 24 to about 30. It is carried out by treating with about 1.1 to about 2 equivalents of arylzin iodide until completion. Such reactions are described, for example, in Rieke, J. Org. Chem. 1991, 56, 1445.

In some cases, the compound of formula (I) or intermediate thereof may include a substituent having one or more sulfur atoms. Such sulfur atoms are, for example, amino acids of formula (II) used in the above reaction may be L-thiazolidine-4-carboxylic acid, L- (5,5-dimethyl) thiazolidine-4-carboxylic acid, L-thiamor Present when derived from polyline-3-carboxylic acid or the like. If present, such sulfur atoms are oxidized before or after the above coupling reaction using conventional reagents and reaction conditions to provide sulfoxide or sulfone compounds. Suitable reagents for oxidizing sulfide compounds to sulfoxides include, for example, hydrogen peroxide, 3-chloroperoxybenzoic acid (MCPBA), sodium periodate, and the like. The oxidation reaction is typically performed by contacting the sulfide compound with about 0.95 to about 1.1 equivalents of oxidant in an inert diluent such as dichloromethane for about 1 to about 24 hours at a temperature ranging from about -50 to about 75 ° C. The resulting sulfoxide can then be oxidized to the corresponding sulfone by contacting the sulfoxide with at least one equivalent of additional oxidant (eg, hydrogen peroxide, MCPBA, potassium permanganate, etc.). The sulfones can also be prepared directly by contacting sulfides with at least two equivalents, preferably with excess oxidant. This reaction is described in "Advanced Organic Chemistry", 4th Ed., Pp. 1201-1202, Wiley Publisher (1992).

Finally, compounds of formula I, wherein Q is -C (S) NR ^7- , can be prepared using amino thianoic acid derivatives instead of amino acids of formula II in the synthesis procedures described above. Such amino thianoic acid derivatives are described in Shalaky, et al., J. Org. Chem., 61: 9045-9048 (1996); Brain, et al., J. Org. Chem., 62: 3808-3809 (1997).

Pharmaceutical formulation

When compounds of formula (I) and formula (IA) are used as medicaments, they are generally administered in the form of pharmaceutical compositions. These compounds can be administered by various routes of administration, such as oral, rectal, transdermal, subcutaneous, intravenous, intramuscular and intranasal. These compounds are effective as both injectable and oral compositions. Such compositions are prepared by methods well known in the art of pharmacy, and comprise one or more active compounds.

The present invention also encompasses pharmaceutical compositions containing, as active ingredient, one or more compounds of Formula (I) and Formula (IA) in association with a pharmaceutically acceptable carrier. In preparing the compositions of the present invention, the active ingredient is generally mixed with excipients and may be diluted into excipients or enclosed in a carrier which may be in the form of capsules, sachets, paper or other containers. If an excipient is used as a diluent, it may be a solid, semisolid or liquid substance which acts as a vehicle, carrier or medium for the active ingredient. Thus, the composition can be, for example, tablets, pills, powders, lozenges, capsules, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or In liquid media), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaging powders.

In formulating a formulation, it may be necessary to grind the active compound to provide particles of the appropriate size prior to combining with other ingredients. If the active compound is substantially insoluble, it is usually milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size is adjusted to be substantially uniformly distributed (eg, about 40 mesh) in the formulation.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, Water, syrup and methyl cellulose. The formulation further comprises lubricants (eg talc, magnesium stearate and mineral oil), wetting agents, emulsifiers and suspending agents, preservatives (methyl- and propylhydroxybenzoate), sweetening agents and flavoring agents. Compositions of the present invention may be formulated to rapidly and sustained or delayed release of the active ingredient after administration to a patient using procedures known in the art.

The compositions are preferably formulated in unit dosage forms each containing a dosage of about 5 to about 100 mg, more generally about 10 to about 30 mg of the active ingredient. The term “unit dosage form” refers to a unit dose containing a predetermined amount of active substance calculated so that each unit, together with a suitable pharmaceutical excipient, will produce the desired therapeutic effect for human subjects and other mammals. It means a physical distinguishing unit suitable for the following.

The active compounds are effective over a wide range of doses and are generally administered in pharmaceutically effective amounts. However, the amount of compound administered substantially is considered to be determined by the physician in appropriate circumstances, including the condition to be treated, the route of administration chosen, the compound administered substantially, the age, weight, sensitivity and severity of the symptom of each patient.

When formulating solid compositions, such as tablets, the main active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compounds of the invention. When these preformulation compositions are referred to as homogeneous, this means that the active ingredients are uniformly dispersed throughout the composition so that they can be easily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. Such solid preformulations are subdivided, for example, in unit dosage forms of the type described above containing from 0.1 to 500 mg of the active ingredient of the invention.

Tablets or pills of the invention provide a dosage form having the advantage of being coated or mixed to act on a continuous basis. For example, a tablet or pill can include an internal dosage component and an external dosage component in a form surrounding the internal dosage component. The two components can be separated by an enteric layer that acts to resist degradation in the stomach and allows the internal components to pass completely into the duodenum or delay release. Various materials can be used as the functional layer or coating, including many polymeric acids and shellac, cetyl alcohol and mixtures of cellulose acetate and polymeric acids.

Liquid forms in which the novel compositions of the present invention may be incorporated for oral or injection administration include elixirs as well as flavor emulsions with liquids, suitable aqueous aqueous syrups, aqueous or oily suspensions and flavor oils with cottonseed oil, sesame oil, coconut oil or peanut oil. And similar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions and suspensions, and powders in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof. The liquid or solid composition may comprise the pharmaceutically acceptable excipients described above. Preferably, the composition is administered orally or by nasal respiratory route for local or systemic effects. Preferably the composition in a pharmaceutically acceptable solvent can be sprayed using an inert gas. The nebulized liquid may be inhaled directly from the nebulization device or the nebulizer may be attached to a face mask tent or an intermittent positive pressure breathing machine. The solution, suspension or powder composition is preferably administered orally or intranasally from a device that carries the formulation in a suitable manner.

The following formulation examples illustrate the pharmaceutical compositions of the present invention.

Formulation Example 1

A hard gelatin capsule containing the following ingredients is prepared:

Ingredient amount (mg / capsule)

Active ingredient 30.0

Starch 305.0

Magnesium Stearate 5.0

The above ingredients are mixed and filled into hard gelatin capsules in an amount of 340 mg.

Formulation Example 2

Tablet formulations are prepared using the following ingredients:

Ingredient amount (mg / tablet)

Active ingredient 25.0

Microcrystalline Cellulose 200.0

Colloidal Silicon Dioxide 10.0

Stearic Acid 5.0

The components are blended and compressed to form tablets each weighing 240 mg.

Formulation Example 3

Anhydrous powder inhalation formulations are prepared comprising the following ingredients:

Component weight%

Active ingredient 5

Lactose 95

The active mixture is mixed with lactose and the mixture is added to an dry powder inhalation instrument.

Formulation Example 4

Tablets each containing 30 mg of the active ingredient are prepared as follows:

Ingredient amount (mg / tablet)

30.0 mg active ingredient

Starch 45.0mg

Microcrystalline Cellulose 35.0mg

Polyvinylpyrrolidone 4.0mg

(As a 10% solution in water)

Sodium carboxymethyl starch 4.5mg

Magnesium Stearate 0.5mg

Talc 1.0mg

120mg total

The active ingredient, starch and cellulose were prepared using No. 20 mesh U.S. Pass through a sieve and mix thoroughly. A solution of polyvinyl-pyrrolidone was mixed with the resulting powder and then No. 16 mesh U.S. Pass it through a sieve. The granules thus obtained were dried at 50-60 ° C. and No. 16 mesh U.S. Pass it through a sieve. 30 mesh U.S. in advance Sodium carboxymethyl starch, magnesium stearate and talc passed through a sieve are added to the granules, followed by mixing and tableting on a tablet machine to give tablets weighing 150 mg each.

Formulation Example 5

Capsules containing 40 mg of drug each are prepared as follows:

Ingredient amount (mg / capsule)

40.0 mg active ingredient

Starch 109.0mg

Magnesium Stearate 1.0mg

150.0 mg total

The active ingredient, cellulose, starch and magnesium stearate are blended and No. 20 Mesh U.S. After passing through a sieve, the hard gelatin capsules are filled in an amount of 150 mg.

Formulation Example 6

Suppositories containing 25 mg of each active ingredient are prepared as follows:

Ingredient amount

25 mg of active ingredient

Make 2,000 mg with saturated fatty acid glycerides.

The active ingredient is No. 60 mesh U.S. Pass through a sieve and suspend in pre-melted saturated fatty acid glycerides using the minimum heat required. The mixture is then poured into a 2.0 g nominal suppository mold and cooled.

Formulation Example 7

Suppositories each containing 50 mg of drug per 5.0 mL dose are prepared as follows:

Ingredient amount

50.0 mg active ingredient

Xanthan Rubber 4.0mg

Sodium carboxymethyl cellulose (11%) /

Microcrystalline Cellulose (89%) 50.0mg

Sucrose 1.75 g

Sodium benzoate 10.0mg

Suitable amount of flavoring and coloring

Make 5 mL of purified water.

Drugs, sucrose and xanthan rubber are blended and No. 10 mesh U.S. After passing through a sieve, it is mixed with a previously prepared solution of microcrystalline cellulose and sodium carboxymethyl cellulose in water. Sodium benzoate, flavors and colorants are diluted with some water and added with stirring. Sufficient water is added to obtain the required volume.

Formulation Example 8

Ingredient amount (mg / capsule)

Active ingredient 15.0 mg

Starch 407.0mg

Magnesium Stearate 3.0mg

Total 425.0mg

The active ingredient, cellulose, starch and magnesium stearate are blended and No. 20 Mesh U.S. After passing through a sieve, the hard gelatin capsules are filled in an amount of 560 mg.

Formulation Example 9

Intravenous formulations are prepared as follows:

Ingredient amount

Active ingredient 250.0 mg

Isotonic saline 1000mL

Formulation Example 10

Topical formulations can be prepared as follows:

Ingredient amount

1 to 10 g of active ingredient

30 g of emulsifying wax

20 g of liquid paraffin

Make 100 g with white soft paraffin.

The white soft paraffin is heated until it melts. Liquid paraffin and emulsifying wax are incorporated and stirred until dissolved. The active ingredient is added and stirred until dispersed. Cool until the mixture solidifies.

Another preferred formulation used in the method of the invention uses a transdermal delivery device ("patch"). Such transdermal patches can be used to provide continuous or discontinuous infusion of the compounds of the invention in controlled amounts. The construction and use of transdermal patches for delivery of a medicament are well known in the art. See, for example, US Pat. No. 5,023,252 (1991.6.11.) Incorporated herein by reference. Such patches can be constructed in a continuous, reactionary fashion or in accordance with the delivery needs of the medicament.

If required or necessary, pharmaceutical agents may be introduced directly or indirectly into the brain. Direct techniques generally involve placing a drug delivery catheter via the blood brain barrier into the ventricle of the host. Such implantable delivery systems for use in delivering biological factors to specific anatomical regions of the body are described in US Pat. No. 5,011,472, which is incorporated herein by reference.

In general, preferred indirect techniques include formulating the composition by converting the hydrophilic agent into a fat soluble agent to provide a drug incubation process. In general, the incubation process is accomplished by blocking the hydroxy, carbonyl, sulfate and primary amino groups present on the drug, making the drug more soluble and making it easier to cross the blood brain barrier. In addition, delivery of hydrophilic agents can be enhanced by intraarterial injection of hypertonic solutions that can temporarily open the blood brain barrier.

Usage

Compounds of the invention can be used to bind VLA-4 (α ₄ β ₁ integrin) in biological samples and thus can be used to assay such samples, for example, for VLA-4. In this assay, the compound is bound on a solid support and a VLA-4 sample is added. The amount of VLA-4 in the sample can be measured by conventional methods, such as sandwich ELISA assays. In addition, labeled VLA-4 can be used in competition assays to determine the presence of VLA-4 in a sample. Other suitable assays are well known in the art.

In addition, certain compounds of the invention inhibit the attachment of leukocytes to endothelial cells mediated by VLA-4 in vivo and thus can be used for the treatment of diseases mediated by VLA-4. Such diseases include inflammatory diseases in mammalian patients such as asthma, Alzheimer's disease, atherosclerosis, AIDS dementia, diabetes (including acute pediatric diabetes), inflammatory bowel disease (including ulcerative colitis and Crohn's disease), multiple sclerosis, Acute leukocyte-mediated lung injury, such as those that occur in rheumatoid arthritis, tissue transplantation, tumor metastasis, meningitis, encephalitis, ataxia and other cerebral trauma, nephritis, retinitis, atopic dermatitis, psoriasis, myocardial ischemia, and adult respiratory distress syndrome Include.

The biological activity of the compounds identified above can be assayed in a variety of systems. For example, the compounds can be immobilized on a solid support to measure the adhesion of cells expressing VLA-4. Using this format, many compounds can be screened. Suitable cells for this assay include all white blood cells known to express VLA-4, such as T cells, B cells, monocytes, eosinophils and basophils. For example, numerous leukocyte cell lines can also be used, including Jurkat and U397.

In addition, test compounds competitively inhibit binding between VLA-4 and VCAM-1, or labeled compounds known to bind VLA-4 and VLA-4, such as compounds of the invention, or antibodies to VLA-4. Test your ability to do this. In these assays, VCAM-1 can be immobilized on a solid surface. In addition, VCAM-1 may be expressed as a recombinant fusion protein with an Ig terminus (eg, IgG) that allows for detection of binding to VLA-4 in an immunoassay. In addition, VCAM-1 expressing cells such as activated endothelial cells or VCAM-1 transfected fibroblasts can be used. For assays that measure the ability to block adhesion to endothelial cells in the brain, the assays described in WO 91/05038 are particularly preferred. This publication is incorporated herein by reference in its entirety.

Many assay formats use labeled assay components. Labeling systems can take many forms. The label can be coupled directly or indirectly to the desired component of the assay according to methods well known in the art. Various marks can be used. The component can be labeled in one of several ways. The most common detection method is the use of ^{radiophotography} with ³ H, ¹²⁵ I, ³⁵ S, ¹⁴ C or ³² P labeled compounds and the like. Non-radioactive labels include antibodies that can act as specific binding pairs for ligands, fluorophores, chemiluminescent agents, enzymes, and labeled ligands that bind the labeled antibody. The choice of label depends on the sensitivity required, the ease of compounding and conjugation, the stability requirements and the available equipment.

In vivo models suitable for demonstrating efficacy in treating an inflammatory response include EAE (experimental autoimmune encephalomyelitis) in mice, rats, guinea pigs or primates, as well as other inflammatory models dependent on α ₄ integrins.

Compounds with the desired biological activity can be modified as needed to provide desirable properties such as improved pharmacological properties (eg, in vivo stability, bioavailability) or detectability in diagnosis. For example, incorporation of one or more D-amino acids in the sulfonamides of the present invention increases in vivo stability. Stability can be assayed by various methods, such as measuring the half-life of a protein during incubation with peptidase or human plasma or serum. Many such protein stability assays are described in Verhoef, et al., Eur. J. Drug Metab. Pharmacokinet., 1990, 15 (2): 83-93.

For diagnostic purposes, various labels may be bound to the compound to provide a detectable signal directly or indirectly. Thus, the compounds of the present invention can be modified in a variety of ways to suit a variety of end purposes while still retaining biological activity. In addition, various reactive sites can be introduced at the ends to bind particles, solid substrates, macromolecules and the like.

Labeled compounds can be used for a variety of in vivo or in vitro applications. Radionuclides (e.g. gamma-emitting radioisotopes such as technetium-99 or indium-111), fluorescent materials (e.g. fluorescein), enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors, chemiluminescent compounds and bioluminescent Various labels such as compounds and the like can be used. One of ordinary skill in the art will recognize other labels suitable for binding to the complex or will be able to identify them using routine experimental methods. Combination of these labels is accomplished using standard techniques common to those skilled in the art.

In vivo applications include diagnostic applications such as monitoring the inflammatory response by detecting the presence of white blood cells expressing VLA-4. In addition, the compounds of the present invention can be used to isolate or label such cells. In addition, as mentioned above, the compounds of the present invention can be used to assay potential inhibitors of VLA-4 / VCAM-1 interactions.

For example, for in vivo diagnostic imaging to identify sites of inflammation, radioisotopes are commonly used according to well known techniques. Radioisotopes can bind to peptides directly or indirectly using intermediate functional groups. For example, chelating agents such as diethylenetriaminepentaacetic acid (DTPA) and ethylenediaminetetraacetic acid (EDTA) and similar molecules are used to bind proteins to metal ion radioisotopes.

Both complexes are magnetic resonance imaging (MRI) or electron spin resonance (ESR), which are well known in the art, and may be labeled with paramagnetic isotopes for in vivo diagnostic purposes. In general, any conventional method for visualizing diagnostic images can be used. In general, gamma and positron emitting radioisotopes are used for camera imaging and paramagnetic isotopes are used for MRI. Thus, the compounds can be used to monitor the process of alleviating inflammatory responses in a subject. By measuring the increase or decrease in lymphocytes expressing VLA-4, one can determine whether a particular therapeutic regimen aimed at alleviating the disease is effective.

The pharmaceutical compositions of the present invention can be used to block or inhibit intracellular adhesions associated with myriad diseases and disorders. For example, many inflammatory disorders are associated with integrins or leukocytes. Treatable disorders include graft rejection (eg, allograft rejection), Alzheimer's disease, atherosclerosis, AIDS dementia, diabetes (including acute childhood diabetes), retinitis, cancer metastasis, rheumatoid arthritis, acute leukocyte-mediated lung injury (eg Adult respiratory distress syndrome), asthma, nephritis, and acute and chronic inflammation (eg, atopic dermatitis, psoriasis, myocardial ischemia) and inflammatory bowel disease (including ulcerative colitis and Crohn's disease). In a preferred embodiment, the pharmaceutical composition is used to treat inflammatory brain disorders such as multiple sclerosis (MS), viral meningitis and encephalitis.

Inflammatory bowel disease is a generic term for two similar diseases referred to as Crohn's disease and ulcerative colitis. Crohn's disease is an idiopathic chronic ulcerative inflammatory disease, characterized by granulomatous inflammatory response, with distinct border formation of all layers of the intestinal wall and invasion of conventional transverse walls. Although the disease mostly affects the terminal ileum and / or the colon, any gastrointestinal tract site from mouth to anus may be involved. Ulcerative colitis is an inflammatory response that is greatly limited to the mucous membranes and submucosal tissue of the colon. Lymphocytes and macrophages are present in large numbers in lesions of inflammatory bowel disease, which can contribute to inflammatory damage.

Asthma is characterized by increased responsiveness of bronchial aqueous phase structures to various stimuli that increase paroxysmal narrowing of the bronchial airways. Stimulation releases various inflammatory mediators from IgE coated mast cells, including histamine, eosinophils and neutrophil chemotactic factors, leukotriene, prostaglandins and platelet activating factors. The release of these factors strengthens basophils, eosinophils and neutrophils, causing inflammatory damage.

Atherosclerosis is a disease of arteries (eg, coronary, carotid, aortic and iliac). The underlying lesion, atherosclerosis, consists of thick lesion plaques in the endovascular membrane, with a lipid nucleus and a fibrous cap covering it. Atherosclerosis damages arterial blood flow, weakening the affected arteries. Myocardial infarction and cerebral infarction are the main consequences of this disease. Macrophages and leukocytes are enhanced in atherosclerosis, contributing to inflammatory damage.

Rheumatoid arthritis is a chronic recurrent inflammatory disease that primarily causes damage and destruction of joints. In general, rheumatoid arthritis affects the bovine joints of the hands and feet first, but may then affect the wrists, elbows, ankles, and knees. Arthritis results from the interaction of synovial cells with white blood cells that penetrate the synovial glands of the joint from the circulatory system (Paul, Immunology, 3d ed., Raven Press (1993)).

Another approach for compounds of the invention is to treat organ or transplant rejection mediated by VLA-4. In recent years, the efficacy of surgical techniques for implanting tissues and organs such as skin, kidneys, liver, heart, lungs, pancreas and bone marrow has been significantly improved. Perhaps an important major problem is that in the beneficiary there is no satisfactory drug that causes immunotolerance to the transplanted allograft or organ. When allogeneic cells or organs are transplanted into a host (ie, the donor and the recipient are different individuals of the same kind), the host's immune system initiates an immune response (host versus graft disease) to the transplanted foreign antigen to repair the transplanted tissue. Easy to destroy CD8 ⁺ cells, CD4 cells and monocytes are all involved in rejection of transplanted tissue. Compounds of the invention that bind to α-4 integrins are particularly useful for blocking allogeneic-induced immune responses in beneficiaries and inhibiting their involvement in transplanted tissue and organ destruction. Paul et al., Transplant International 9: 420-425 (1996); Georczynski et al., Immunology 87, 573-580 (1996); Georcyznski et al., Transplant. Immunol. 3: 55-61 (1995); Yang et al., Transplantation 60: 71-76 (1996); Anderson et al., APMIS 102: 23-27 (1994).

A related use of a compound of the invention that binds VLA-4 is to modulate the immune response involved in "graft versus host disease (GVHD). Schlegel et al., J. Immunol. 155: 3856-3865 (1995). GVHD is a potentially fatal disease that occurs when immunologically adapted cells are delivered to allograft recipients. In this situation, the donor's immune adaptive cells will attack the recipient's tissue. Skin, intestinal epidermis and liver tissue are frequently targeted and can be destroyed during GVHD. The disease is a particularly serious problem when immune tissues are implanted (eg bone marrow transplantation), but GVHD is reported to be less severe in other cases, including heart or liver transplantation. The therapeutic agents of the present invention are particularly used to block the activation of donor T-cells and to inhibit their ability to lyse target cells in the host.

A further use of the compounds of the invention is to inhibit tumor metastasis. Several tumor cells have been reported to express VLA-4, and compounds that bind VLA-4 block the attachment of these cells to endothelial cells. Steinback et al., Urol. Res. 23: 175-83 (1995); Orosz et al., Int. J. Cancer 60: 867-71 (1995); Freedman et al., Leuk. Lymphoma 13: 47-52 (1994); Okahara et al., Cancer Res. 54: 3233-6 (1994).

Another use of the compounds of the present invention is to treat multiple sclerosis. Multiple sclerosis is a progressive neuropathic autoimmune disease that affects an estimated 250,000 to 350,000 people in the United States. Multiple sclerosis is thought to be the result of a special autoimmune reaction in which certain white blood cells begin to attack and destroy myelin, an insulating sheath covering nerve fibers. In multiple sclerosis animal models, murine monoclonal antibodies against VLA-4 have been shown to block the attachment of leukocytes to the endothelium, thereby inhibiting inflammation of the central nervous system in animals, thereby inhibiting subsequent paralysis ¹⁶ .

The pharmaceutical compositions of the invention are suitable for use in a variety of drug delivery systems. Formulations suitable for use in the present invention can be found in the literature (Remington's Pharmaceutical Sciences, Mace Publishing Company, Philadelphia, PA, 17th ed. (1985)).

To enhance serum half-life, the compounds can be trapped in liposomes, introduced into the lumens of liposomes, prepared as colloids, or other conventional techniques can be used to provide extended serum half-life of the compounds. For example, various techniques may be used to prepare liposomes described in US Pat. Nos. 4,235,871, US Pat. No. 4,501,728, and US Pat. No. 4,837,028 to Szoka et al., Each of which is incorporated herein by reference. have.

The amount administered to the patient may vary depending on the substance to be administered, the purpose of administration (eg, prophylactic or therapeutic), the condition of the patient, the method of administration and the like. In therapeutic applications, the composition is administered to a patient suffering from the disease in an amount sufficient to treat or at least partially arrest the symptoms of the disease and its complications. An amount sufficient to achieve this is defined as "therapeutically effective amount". The amount effective for this use depends on the judgment of the attending physician, which depends not only on the condition of the disease to be treated but also on the severity of inflammation, the age, weight and general condition of the patient.

The composition administered to the patient is in the pharmaceutical form described above. Such compositions may be sterilized or sterile filtered by conventional sterilization techniques. The resulting aqueous solution is packaged or lyophilized for use and the lyophilized formulation is mixed with a sterile aqueous carrier prior to administration. The pH of the compound formulation is typically 3 to 11, more preferably 5 to 9, most preferably 7 to 8. It will be appreciated that the specific excipients, carriers or stabilizers described above will be used to produce the form of the pharmaceutical salt.

The therapeutic dosage of a compound of the present invention may vary depending on, for example, the particular use for which the treatment is to be made, the method of administering the compound, the health and condition of the patient, and the judgment of the prescribing physician. For example, for intravenous administration, the dosage is usually in the range of about 20 to about 500 μg, preferably about 100 to about 300 μg, per kilogram of body weight. For intranasal administration, a suitable dosage range is generally about 0.1 pg to 1 mg per kilogram of body weight. Effective dosages can be extrapolated from dose-response curves derived from in vivo or animal model test systems.

The following synthetic and biological examples are provided for the purpose of illustrating the invention and should not be construed as limiting the scope of the invention. Unless otherwise noted, all temperatures are in degrees Celsius.

Example

In the following examples, the following abbreviations have the following meanings. If an abbreviation is not defined, it is generally accepted meaning.

aq or aq. = Aqueous

AcOH = acetic acid

bd = broad doublet

bm = broad multiline

bs = broad single line

Bn = benzyl

Boc = N-t-butoxylcarbonyl

Boc ₂ O = di-t-butyl dicarbonate

BOP = benzotriazol-1-yloxy-tris (dimethylamino) phosphonium

Hexafluorophosphate

Cbz = carbobenzyloxy

CHCl ₃ = chloroform

CH ₂ Cl ₂ = dichloromethane

(COCl) ₂ = oxalyl chloride

d = doublet

dd = doublet of doublet

dt = doublet of triplets

DBU = 1,8-diazabicyclo [5.4.0] undec-7-yen

DCC = 1,3-dicyclohexylcarbodiimide

DMAP = 4-N, N-dimethylaminopyridine

DME = ethylene glycol dimethyl ether

DMF = N, N-dimethylformamide

DMSO = dimethyl sulfoxide

EDC = 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide

Hydrochloride

Et ₃ N = triethylamine

Et ₂ O = diethyl ether

EtOAc = ethyl acetate

EtOH = ethanol

eq or eq. = Equivalent

Fmoc = N- (9-fluorenylmethoxycarbonyl)

FmocONSu = N- (9-fluorenylmethoxycarbonyl) -succinimide

g = grams

h = hours

H ₂ O = water

HBr = bromic acid

HCl = hydrochloric acid

HOBT = 1-hydroxybenzotriazole anhydride

hr = hour

K ₂ CO ₃ = potassium carbonate

L = liter

m = polyline

MeOH = Methanol

mg = milligrams

MgSO ₄ = magnesium sulfate

mL = milliliters

mm = millimeters

mM = millimolar concentration

mmol = millimoles

mp = melting point

N = normal concentration

NaCl = Sodium Chloride

Na ₂ CO ₃ = sodium carbonate

NaHCO ₃ = sodium bicarbonate

NaOEt = Sodium Ethoxide

NaOH = sodium hydroxide

NH ₄ Cl = ammonium chloride

NMM = N-methylmorpholine

Phe = L-phenylalanine

Pro = L-Proline

psi = pounds per square inch

PtO ₂ = platinum oxide

q = quadruple

q.s. = Sufficient

quint. = Quintet

rt = room temperature

s = single line

sat = saturated

t = triplet

t-BuOH = t-butanol

TFA = trifluoroacetic acid

THF = tetrahydrofuran

TLC or tlc = thin layer chromatography

Ts = tossile

TsCl = tosyl chloride

TsOH = tosylate

μL = microliters

In the examples below, all temperatures are in degrees Celsius unless otherwise noted. The following compounds are prepared using the following method as shown below.

Method 1

N-Tosylation Method

The N-tosylation reaction of the appropriate amino acid is carried out by the method of literature. Cupps. Boutin and Rapoport, J. Org. Chem. 1985, 50: 3972].

Method 2

Method for preparing methyl ester

Amino acid methyl esters are prepared using the methods in the literature. Brenner and Huber, Helv. Chim. Acta, 1953, 36: 1109.

Method 3

BOP Coupling Method

The desired dipeptide esters can be prepared by reacting a suitable N-protected amino acid (1 equivalent) with an appropriate amino acid ester, or by amino acid ester hydrochloride (1 equivalent), benzotriazol-1-yloxy-tris (dimethylamino) phosphonium Prepared by reaction with hexafluorophosphate [BOP] (2.0 equiv), triethylamine (1.1 equiv) and DMF. The reaction mixture is stirred overnight at room temperature. Crude product is purified by flash chromatography to yield the dipeptide ester.

Method 4

Hydrogenation Method I

The hydrogenation reaction is carried out using 10% Pd / C (10% by weight) at 30 psi in methanol. The mixture is filtered through a pad of celite and concentrated to afford the desired amino compound.

Method 5

Hydrolysis Method I

LiOH (or NaOH) (0.95 equiv) is added to a cooled (0 ° C.) THF / H ₂ O solution (2: 1, 5-10 mL) of the appropriate ester. The temperature is maintained at 0 ° C. and the reaction completes in 1 to 3 hours. The reaction mixture is extracted with ethyl acetate and the aqueous phase is lyophilized to give the desired carboxylate salt.

Method 6

Ester Hydrolysis Method II

LiOH (1.1 equiv) is added to a cooled (0 ° C.) THF / H ₂ O solution (2: 1, 5-10 mL) of the appropriate ester. The temperature is maintained at 0 ° C. and the reaction completes in 1 to 3 hours. The reaction mixture is concentrated and the residue is worked up in H ₂ O and then the pH is adjusted to 2-3 with aqueous HCl. The product is extracted with ethyl acetate and the combined organic phases are washed with brine, then dried over MgSO ₄ , filtered and then concentrated to give the desired acid.

Method 7

Ester Hydrolysis Method III

The appropriate ester is dissolved in dioxane / H ₂ O solution (1: 1) and 0.9 equivalent of 0.5N NaOH is added. The reaction is stirred for 3-16 hours and then concentrated. The resulting residue is dissolved in H ₂ O and extracted with ethyl acetate. The aqueous phase is lyophilized to give the desired carboxylate sodium salt.

Method 8

Sulfonylation Method I

To the appropriately protected aminophenylalanine homologue (11.2 mmol) dissolved in methylene chloride (25 mL) and cooled to -78 ° C, the desired sulfonyl chloride (12 mmol) is added, followed by the dropwise addition of pyridine (2 mL). The solution is warmed to rt and stirred for 48 h. The reaction solution is added with methylene chloride (100 mL) to a 250 mL separatory funnel and extracted with 1N HCl (50 mL × 3), brine (50 mL) and water (100 mL). The organic phase is dried (MgSO ₄ ) and the solvent is concentrated to give the desired product.

Method 9

Reductive Amination Method

Reductive amination of Tos-Pro-p-NH ₂ -Phe with the appropriate aldehyde is carried out using acetic acid, sodium triacetoxyborohydride and methylene chloride, and the combined mixture is stirred overnight at room temperature. Flash chromatography purifies the crude product.

Method 10

How to remove a BOC

Anhydrous hydrochloric acid (HCl) gas is bubbled through an appropriate methanol solution of Boc-amino acid ester at 0 ° C. for 15 minutes and the reaction mixture is stirred for 3 hours. The solution is concentrated in syrup, dissolved in Et ₂ O and then concentrated again. This process is repeated and the resulting solid is left overnight under high vacuum.

Method 11

t-butyl ester hydrolysis method I

t-butyl ester is dissolved in CH ₂ Cl ₂ and treated with TFA. The reaction is completed within 1 to 3 hours when the reaction mixture is concentrated, and the residue is dissolved in H ₂ O and lyophilized to give the desired acid.

Method 12

EDC Coupling Method I

Amino acid ester hydrochloride (1 equivalent), N-methylmorpholine (1.1-2.2 equivalents), suitable for a CH ₂ Cl ₂ solution (5-20 mL) of N- (toluene-4-sulfonyl) -L-proline (1 equivalent) ) And 1-hydroxybenzotriazole (2 equiv) are added and mixed, left in an ice bath, and then 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide (1.1 equiv) is added. The reaction is left to rise to room temperature and stirred overnight. The reaction mixture is poured into H ₂ O, washed with saturated NaHCO ₃ and brine, dried over MgSO ₄ or Na ₂ SO ₄ , filtered and concentrated. The crude product is purified by column chromatography.

Method 13

EDC Coupling Method II

To the appropriate DMF solution of N-protected amino acid (5-20 mL) add the appropriate amino acid ester hydrochloride (1 equiv), Et ₃ N (1.1 equiv) and 1-hydroxybenzotriazole (2 equiv) and ice bath It is left to and 1- (3-dimethylaminopropyl) -3-ethyl carbodiimide (1.1 equiv) is added. The reaction is left to rise to room temperature and stirred overnight. The reaction mixture is partitioned between EtOAc and H ₂ O and the organic phase is washed with 0.2N citric acid, H ₂ O, saturated NaHCO ₃ and brine, dried over MgSO ₄ or Na ₂ SO ₄ , filtered and concentrated. Crude product is purified by column chromatography or preparative TLC.

Method 14

Sulfonylation Method II

Appropriate sulfonyl chloride is dissolved in CH ₂ Cl ₂ and left in an ice bath. L-Pro-L-Phe-OMe.HCl (1 equiv) and Et ₃ N (1.1 equiv) are added, the reaction is left to warm to room temperature and stirred overnight under a nitrogen atmosphere. The reaction mixture is concentrated and the residue is partitioned between EtOAc and H ₂ O, the organic phase is washed with saturated NaHCO ₃ and brine, dried over MgSO ₄ or Na ₂ SO ₄ , filtered and concentrated. Crude product is purified by column chromatography or preparative TLC.

Method 15

Sulfonylation Method III

Et ₃ N (produced using the process described in Method 10) of L-Pro-L-4- (3-dimethylaminopropyloxy) -Phe-OMe in CH ₂ Cl ₂ (prepared using the process described in Method 10) 5 equivalents) is added followed by the appropriate sulfonyl chloride (1.1 equiv). The reaction is left to warm to room temperature and stirred overnight under a nitrogen atmosphere. The mixture is concentrated, dissolved in EtOAc and washed with saturated NaHCO ₃ and 0.2N citric acid. The aqueous phase is basified with solid NaHCO ₃ and the product is extracted with EtOAc. The organic phase is washed with brine, dried over MgSO ₄ or Na ₂ SO ₄ , filtered and concentrated. Crude methyl esters are purified by preparative TLC. The corresponding acid is prepared using the process described in Method 7.

Method 16

Hydrogenation Method II

To a methanol solution of azlactone (10-15 mL) add NaOAc (1 equiv) and 10% Pd / C. This mixture is left in a hydrogenator of H ₂ 40 psi. After 8-16 hours, the reaction mixture is filtered through a pad of celite and the filtrate is concentrated to give the dehydrodipeptide methyl ester. The ester is dissolved in dioxane / H ₂ O (5-10 mL) and 0.5N NaOH (1.05 equiv) is added. It is stirred for 1-3 hours, then the reaction mixture is concentrated and the residue is redissolved in H ₂ O and washed with EtOAc. The aqueous phase is acidified with 0.2N HCl and the product is extracted with EtOAc. The combined organic phases are washed with brine (1 × 5 mL), dried over MgSO ₄ or Na ₂ SO ₄ , filtered and concentrated to give the acid as an about 1: 1 mixture of diastereomers.

Method 17

t-butyl ester hydrolysis method II

t-butyl ester is dissolved in CH ₂ Cl ₂ (5 mL) and treated with TFA (5 mL). The reaction is complete within 1 to 3 hours when the reaction mixture is concentrated, the residue is dissolved in H ₂ O and then concentrated. The residue is redissolved in H ₂ O and lyophilized to give the desired product.

Example 1

Synthesis of N- (methanesulfonyl) -L-prolyl-L-phenylalanine

Boc-L-Pro-OH and L-Phe-OBn.HCl are treated with BOP and NMM in DMF, and after aqueous workup and flash chromatography, Boc-L-Pro-OBn is obtained. This product is then treated with TFA and anisol and the mixture is distilled off. The residue is dissolved with Et ₂ O and washed with saturated aqueous NaHCO ₃ and saturated aqueous NaCl, the Et ₂ O layer is dried over anhydrous MgSO ₄ , filtered and the solvent is distilled off to give L-Pro-L- Obtain Phe-OBn. The residue was treated with CH ₃ SO ₂ Cl and Et ₃ N in CH ₂ Cl ₂ , aqueous post-treatment and flash chromatography followed by N- (CH ₃ SO ₂ ) -L-Pro-L-Phe-OBn. To obtain. This product is then treated with 10% Pd / C in THF and then the mixture is stirred under 50 psi of H ₂ . The mixture is filtered through celite and distilled to afford the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 7.93 (d, J = 8.2, 1H), 7.27-7.13 (m, 5H), 4.51-4.45 (m, 1H), 4.15 (dd, J = 8.7 , J = 2.6, 1H), 3.59 (t, J = 6.1, 1H), 3.30-3.25 (m, 2H), 3.10 (dd, J = 13.7, J = 4.9, 1H), 2.96 (dd, J = 13.7 , J = 9.1, 1H), 2.87 (s, 3H), 2.03-1.93 (m, 1H), 1.77-1.54 (m, 3H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 171.1, 137.4, 129.2, 128.1, 126.4, 61.2, 53.0, 48.6, 36.5, 35.2, 30.6, 24.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 341 (MH &lt; + &gt;).

Example 2

Synthesis of N- (α-toluenesulfonyl) -L-prolyl-L-phenylalanine

CH ₃ SO ₂ Cl is substituted with PhCH ₂ SO ₂ Cl and by the preparation of Example 1 (9) the title compound is obtained as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.01 (d, J = 8.2, 1H), 7.39-7.20 (m, 5H), 7.18-7.13 (m, 5H), 4.53-4.46 (m, 1H ), 4.42 (d, J = 13.5, 1H), 4.35 (d, J = 13.5, 1H), 4.20-4.17 (m, 1H), 3.31-3.18 (m, 3H), 3.09 (dd, J = 13.9, J = 4.8, 1H), 2.96 (dd, J = 13.8, J = 8.9, 1H), 2.03-1.95 (m, 1H), 1.79-1.58 (m, 3H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 171.5, 137.6, 131.0, 129.3, 129.0, 128.9, 128.1, 128.0, 126.3, 61.5, 53.3, 48.8, 36.5, 30.8, 30.6, 24.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 417 (MH &lt; + &gt;).

Example 3

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine

CH ₃ SO ₂ Cl is substituted with 4-CH ₃ (C ₆ H ₄ ) SO ₂ Cl and by the preparation of Example 1 (9) the title compound is obtained as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.07 (d, J = 8.0, 1H), 7.68 (d, J = 8.2, 2H), 7.39 (d, J = 8.1, 2H), 7.29-7.15 (m, 5H), 4.52-4.44 (m, 1H), 4.10-4.07 (m, 1H), 3.34-3.27 (m, 1H), 3.12-3.06 (m, 2H), 2.98 (dd, J = 13.7, J = 8.7, 1 H), 2.39 (s, 3 H), 1.57-1.36 (m, 4H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 170.8, 143.6, 137.4, 133.8, 129.8, 129.3, 128.1, 127.5, 126.4, 61.3, 53.2, 49.0, 36.6, 30.4, 23.7, 21.0.

Mass spectroscopy: (+ FAB, Glycerol / Trifluoroacetic acid) 417 (MH &lt; + &gt;).

Example 4

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L- (N-methyl) phenylalanine

N-methyl-L-Phe-OH is treated with benzyl alcohol and 4-methylphenylsulfonic acid to give N-methyl-L-Phe-OBnTsOH. Substitute D-Pro-OH with L-Pro-OH and L-Phe-OBn.TsOH with N-methyl-Phe-OBn.TsOH. Is obtained as a clean oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 7.60 (d, J = 8.2, 2H), 7.34 (d, J = 8.2, 2H), 7.27-7.16 (m, 5H), 4.97-4.92 (m) , 1H), 4.64-4.61 (m, 1H), 3.33 (bs, 1H), 3.25-3.12 (m, 3H), 3.04 (dd, J = 10.5, J = 14.6, 1H), 2.85 (s, 3H) , 2.37 (s, 3H), 1.86-1.72 (m, 2H), 1.68-1.50 (m, 2H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 171.9, 171.1, 143.0, 137.9, 135.6, 129.6, 128.7, 128.2, 127.2, 126.3, 58.6, 48.0, 33.4, 33.0, 30.4, 29.6, 23.9, 21.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 431 (MH &lt; + &gt;).

Example 5

Synthesis of N- (toluene-4-sulfonyl) -L-pipecolinyl-L-phenylalanine

D-Pro-OH was substituted with L-pipecolinic acid and the preparation of Example 6 (17) gave the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.09 (d, J = 8.0, 1H), 7.44 (d, J = 8.2, 2H), 7.34-7.21 (m, 5H), 7.17 (d, J = 8.2, 2H), 4.44 (d, J = 4.8, 1H), 4.32-4.24 (m, 1H), 3.61-3.53 (m, 2H), 3.33 (bs, 1H), 3.17-3.11 (m, 1H) , 3.07 (dd, J = 13.8, J = 4.5, 1H), 2.89 (dd, J = 13.8, J = 9.9, 1H), 2.33 (s, 3H), 1.88 (bd, J = 12.9, 1H), 1.40 -1.30 (m, 3H), 1.08-1.05 (m, 2H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.8, 169.8, 142.7, 137.7, 136.6, 129.4, 129.2, 128.2, 126.8, 126.5, 54.1, 53.4, 42.4, 36.3, 30.4, 26.7, 23.5, 21.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 431 (MH &lt; + &gt;).

Example 6

Synthesis of N- (toluene-4-sulfonyl) -D-prolyl-L-phenylalanine

D-Pro-OH is treated with TsCl and NaOH in H ₂ O, oxidized, extracted, dried over anhydrous MgSO ₄ , evaporated and N- (toluene-4-sulfonyl) -D-Pro-OH is obtained. . This product was treated with H-Phe-OBn.HCl, BOP and NMM in DMF, after aqueous workup and flash chromatography, to yield N- (toluene-4-sulfonyl) -D-Pro-Phe-OBn. do. This product is treated with 10% Pd / C in T HF and the mixture is stirred under 50 psi of H ₂ . The mixture is filtered through celite and distilled to afford the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.10 (d, J = 8.4, 1H), 7.70 (d, J = 8.2, 2H), 7.40 (d, J = 8.2, 2H), 7.28-7.15 (m, 5H), 4.51-4.44 (m, 1H), 4.13 (t, J = 5.7, 1H), 3.33-3.25 (m, 2H), 3.11-3.03 (m, 2H), 2.94 (dd, J = 13.8, J = 9.0, 1H), 2.39 (s, 3H), 1.66-1.57 (m, 1H), 1.50-1.37 (m, 3H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 170.8, 143.5, 137.3, 134.1, 129.8, 129.3, 128.1, 127.4, 126.4, 61.3, 53.0, 48.9, 36.8, 30.4, 23.8, 21.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 417 (MH &lt; + &gt;).

Example 7

Synthesis of N- (toluene-4-sulfonyl) -L- (trans-4-hydroxy) -prolyl-L-phenylalanine

D-Pro-OH was substituted with trans-4-hydroxy-L-proline and the preparation of Example 6 (17) gave the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.25 (d, J = 8.1, 1H), 7.64 (d, J = 8.2, 2H), 7.36 (d, J = 8.2, 2H), 7.30-7.15 (m, 5H), 4.81 (bs, 1H), 4.47-4.40 (m, 1H), 4.15-4.10 (m, 2H), 3.44 (dd, J = 10.3, J = 4.8, 1H), 3.35 (bs, 1H), 3.09-2.91 (m, 3H), 2.38 (s, 3H), 1.72-1.64 (m, 2H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 171.0, 143.1, 137.4, 134.1, 129.5, 129.3, 128.1, 127.7, 126.4, 68.1, 60.3, 56.1, 53.4, 36.7, 30.4, 21.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 433 (MH &lt; + &gt;).

Example 8

Synthesis of N- (toluene-4-sulfonyl) -L- (azetidine-2-carbonyl) -L-phenylalanine

D-Pro-OH is substituted with (S) -2-azetidinecarboxylic acid and the preparation of Example 6 (17) affords the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.12 (d, J = 8.0, 1H), 7.69 (d, J = 8.2, 2H), 7.47 (d, J = 8.2, 2H), 7.31-7.13 (m, 5H), 4.54-4.47 (m, 1H), 4.23 (t, J = 8.3, 1H), 3.67-3.58 (m, 2H), 3.47 (q, J = 8.4, 1H), 3.33 (bs, 1H), 3.10 (dd, J = 13.7, J = 5.2, 1H), 2.98 (dd, J = 13.6, J = 7.9, 1H), 2.42 (s, 3H), 2.03-1.87 (m, 2H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.3, 168.7, 144.3, 137.1, 130.9, 130.0, 129.4, 128.3, 128.2, 126.6, 61.8, 53.1, 47.9, 36.7, 21.1, 19.6.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 403 (MH &lt; + &gt;).

Example 9

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-D, L-homophenylalanine

N- (toluene-4-sulfonyl) -L-proline (250 mg, 0.92 mmol) was converted to D, L-homophenylalanine methyl ester (166 mg, 1.1 equiv), Et ₃ N (2.1 equiv, 283 μL) and BOP (1.1 equiv) 410 mg) in DMF (20 mL). Dipeptide is isolated as an oil in 73% yield (300 mg, 0.67 mmol). The ester (300 mg, 0.67 mmol) is then hydrolyzed in a 1: 1 MeOH: H ₂ O (5 mL) solution with NaOH (1.1 equiv, 28 mg). The acid is separated off as a foam in a yield of 60%.

NMR data are as follows:

¹ H NMR (300 MHz, CD ₃ OD): δ = 7.60 (m, 2H), 7.20 (m, 2H), 7.03 (m, 5H), 4.23-3.87 (m, 2H), 3.42 (m, 1H), 3.29 (m, 1 H), 2.58 (m, 2 H), 2.20 (s, 3 H), 2.02-1.38 (m, 7H).

¹³ C NMR (75 MHz, CD ₃ OD): δ = 175.53, 175.17, 146.49, 146.19, 142.93, 135.91, 135.04, 131.72, 130.28, 129.61, 129.49, 129.21, 127.70, 64.39, 63.59, 53.76, 53.52, 51.51 , 35.15, 33.42, 32.74, 32.60, 32.48, 26.28, 26.20, 22.14.

Mass spectroscopy: (FAB) 431 (M + H).

Example 10

Synthesis of N- (4-chlorobenzenesulfonyl) -L-prolyl-L-phenylalanine

A CH ₃ SO ₂ Cl by the production process of 4-chloro-phenyl and optionally substituted with -SO ₂ Cl, in Example 1 (9), to give the title compound having a melting point of 69 to 71 ℃ as a solid.

NMR data are as follows:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.77 (m, 2H), 7.53 (m, 2H), 7.31 (m, 5H), 4.72 (m, 1H), 4.04 (m, 1H), 3.35 (m , 2H), 3.10 (m, 2H), 2.05 (m, 1H), 1.54 (m, 3H).

¹³ C NMR (75 MHz, CD ₃ OD): δ = 174.67, 174.41, 141.23, 138.81, 137.55, 135.02, 131.18, 130.04, 129.45, 128.45, 63.75, 55.48, 38.79, 32.32, 25.87 (1C buried under solvent peak)

Mass spectroscopy: (FAB) 437 (M + H).

Example 11

Synthesis of N- (1-naphthalenesulfonyl) -L-prolyl-L-phenylalanine

A CH ₃ SO ₂ Cl by the method of Example 1 (9) substituted, 1-naphthyl and to -SO ₂ Cl, to give the title compound as a solid.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.73-8.70 (m, 1H), 8.24 (d, J = 8.2, 1H), 8.12-8.06 (m, 2H), 7.69-7.57 (m, 2H ), 7.33-7.13 (m, 7H), 4.40-4.33 (m, 2H), 3.34-3.23 (m, 3H), 3.03 (dd, J = 14.0, J = 5.2, 1H), 2.88 (dd, J = 13.7, J = 8.8, 1 H), 1.79-1.15 (m, 4 H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 170.9, 137.4, 134.3, 134.0, 133.6, 129.2, 129.1, 129.0, 128.23, 128.18, 128.1, 126.9, 126.4, 124.7, 124.6, 60.6, 53.4, 48.7, 36.5, 30.8, 24.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 475 (MNa +).

Example 12

Synthesis of N- (2-naphthalenesulfonyl) -L-prolyl-L-phenylalanine

A CH ₃ SO ₂ Cl by the method of Example 1 (9) substituted, and 2-naphthyl, -SO ₂ Cl, to give the title compound as a solid.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.18-8.03 (m, 4H), 7.82 (dd, J = 8.7, J = 1.8, 1H), 7.74-7.64 (m, 2H), 7.32-7.15 (m, 6H), 4.53-4.46 (m, 1H), 4.25-4.21 (m, 1H), 3.34-3.27 (m, 3H), 3.10 (dd, J = 13.8, J = 5.1, 1H), 2.99 ( dd, J = 13.7, J = 8.8, 1H), 1.56-1.36 (m, 4H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 170.8, 137.4, 134.4, 133.9, 131.8, 129.4, 129.3, 129.0, 128.7, 128.1, 127.8, 127.6, 127.4, 122.8, 61.4, 53.2, 49.1, 36.6, 30.4, 23.8.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 475 (MNa +).

Example 13

Synthesis of N- (4-methoxybenzenesulfonyl) -L-prolyl-L-phenylalanine

A CH ₃ SO ₂ Cl 4- methoxyphenyl the production method of the -SO ₂ Cl was replaced by, in Example 1 (9), to give the title compound as a solid.

NMR data are as follows:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.77 (m, 2H), 7.31 (m, 5H), 7.01 (m, 2H), 4.80 (m, 1H), 4.05 (m, 1H), 3.86 (s , 3H), 3.35 (m, 2H), 3.10 (m, 2H), 1.96 (m, 1H), 1.54 (m, 3H).

¹³ C NMR (75 MHz, CD ₃ OD): δ = 174.69, 174.54, 165.63, 136.75, 131.69, 131.07, 130.02, 128.45, 116.13, 63.84, 56.83, 55.35, 51.09, 38.81, 32.15, 26.63.

Mass spectroscopy: (FAB) 433 (M + H).

Example 14

Synthesis of N- (4-t-butylbenzenesulfonyl) -L-prolyl-L-phenylalanine

A CH ₃ SO ₂ Cl by the production process of 4-t- butylphenyl -SO was substituted with Cl _2, Example 1 (9), to give the title compound as a solid.

NMR data are as follows:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.75 (d, 2H, J = 8.52 Hz), 7.55 (d, 2H, J = 8.43 Hz), 7.29 (m, 5H), 4.80 (m, 1H), 4.05 (m, 1H), 3.30 (m, 3H), 3.10 (m, 3H), 1.95 (m, 1H), 1.56 (m, 2H), 1.34 (s, 9H).

¹³ C NMR (75 MHz, CD ₃ OD): δ = 174.81, 174.47, 159.01, 138.79, 135.59, 131.09, 130.02, 129.45, 128.43, 128.05, 63.83, 55.46, 51.09, 38.84, 36.66, 32.20, 31.99, 25.83.

Mass spectroscopy: (FAB) 459 (M + H).

Example 15

Synthesis of N- (toluene-4-sulfonyl) -L- (trans-4-fluoro) -prolyl-L-phenylalanine

The product from Example 49 (176) is treated with 10% Pd / C in THF and the mixture is stirred under 50 psi of H ₂ . The mixture is filtered through celite and distilled to afford the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.43 (d, J = 8.1, 1H), 7.66 (d, J = 8.3, 2H), 7.35 (d, J = 8.3, 2H), 7.29-7.13 (m, 5H), 5.10 (bd, J = 53.3, 1H), 4.49-4.42 (m, 1H), 4.19 (t, J = 8.3, 1H), 3.63-3.46 (m, 2H), 3.34 (bs, 1H), 3.06 (dd, J = 13.7, J = 5.4, 1H), 2.95 (dd, J = 13.7, J = 8.3, 1H), 2.38 (s, 3H), 2.25-2.12 (m, 1H), 1.94 -1.72 (m, 1 H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.5, 170.4, 143.4, 137.4, 134.2, 129.5, 129.4, 128.1, 127.6, 126.5, 92.0 (d, J = 177 Hz), 59.9, 55.1, 53.4, 37.6 (d, J = 21 Hz), 36.8, 21.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 435 (MH &lt; + &gt;).

Example 16

Synthesis of N- (toluene-4-sulfonyl) -L- (cis-4-fluoro) -prolyl-L-phenylalanine

The product from Example 50 (177) is treated with 10% Pd / C in THF and the mixture is stirred under H ₂ 50 psi. The mixture is filtered through celite and distilled to afford the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 7.78 (d, J = 7.7, 1H), 7.73 (d, J = 8.2, 2H), 7.41 (d, J = 8.2, 2H), 7.29-7.13 (m, 5H), 5.10 (dt, Jd = 52.9, Jt = 3.5, 1H), 4.53-4.46 (m, 1H), 4.31-4.27 (m, 1H), 3.60-3.28 (m, 3H), 3.06- 2.94 (m, 2H), 2.39 (s, 3H), 2.18 (dd, J = 19.8, J = 15.2, 1H), 1.93-1.70 (m, 1H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.1, 169.7, 144.1, 136.8, 132.8, 130.0, 129.3, 128.1, 127.7, 126.6, 92.0 (d, J = 176 Hz), 60.5, 55.1 (d, J = 23.5), 53.2, 36.9, 36.2, 21.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 435 (MH &lt; + &gt;).

Example 17

Synthesis of N- (toluene-4-sulfonyl) -L- (5,5-dimethyl) -thiaprolyl-L-phenylalanine

N- (toluene-4-sulfonyl) -L- (5,5-dimethyl) -thiazolidine-4-carboxylic acid was converted to L- (5,5-dimethyl) -thiazolidine using the procedure described in Method 1. Prepared from -4-carboxylic acid. The title compound is prepared following the procedure described for Example 1 (9) to afford a solid having a melting point of 79 to 81 ° C.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 0.98 (s, 3H), 1.08 (s, 3H), 2.42 (s, 3H), 3.04-3.34 (m, 2H), 3.88 (s, 1H), 4.38 (d, 1H, J = 10.3 Hz), 4.52 (d, 1H, J = 10.2 Hz), 4.90 (m, 1H), 7.09 (bd, 1H), 7.15-7.37 (m, 7H), 7.73 (d, 2H, J = 8.2 Hz).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 22.3, 24.3, 29.6, 38.3, 51.2, 54.1, 55.3, 73.9, 127.7, 128.7, 129.2, 130.1, 130.7, 133.0, 136.5, 145.6, 169.9, 174.5.

Mass spectroscopy: (FAB +) 463 (M + H).

Example 18

Synthesis of N- (2-methoxycarbonylbenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound having a melting point of 163 to 165 ° C. is prepared as a yellow solid from the product of Example 53 using the procedure described in Method 4.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 8.74 (bs, 1H), 7.93 (d, 1H, J = 7.57 Hz), 7.70 (d, 1H, J = 8.12 Hz), 7.57 & 7.25 (M, 8H ), 4.69 (m, 1H), 4.39 (d, 1H, J = 7.69 Hz), 3.93 (s, 3H), 3.44 (m, 1H), 3.29 (m, 2H), 2.93 (m, 1H), 1.88 , 1.62 & 1.17 (m, 4 H).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 175.67, 172.45, 169.27, 137.19, 135.39, 134.12, 133.67, 131.42, 130.55, 129.77, 129.52, 129.09, 127.52, 62.19, 54.08, 49.34, 37.58, 31.48, 30 24.21.

Mass spectroscopy: (+ FAB) 461 (M + H) 483 (M + Na).

Example 19

Synthesis of N- (2-carboxybenzenesulfonyl) -L-prolyl-L-phenylalanine

The product of Example 18 (56) (1.8 mmol) is mixed with dioxane (25 mL) and 1N NaOH (1.8 mmol) and the reaction is stirred at rt for 16 h. 1N HCl (1.8 mmol) is added and water and dioxane are removed under reduced pressure. The product is extracted with 0.2N NaOH (50 mL) and EtOAc (2 × 50 mL) and the recovered organic layer is dried over MgSO ₄ , filtered and concentrated to give a colorless oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 11.15 (bs, 2H), 7.95, 7.57 & 7.21 (m, 9H), 4.74 (m, 1H), 4.53 (m, 1H), 3.44 (m, 1H) , 3.28 (m, 2H), 2.91 (m, 1H), 1.89 (M, 2H), 1.60 (m, 1H), 1.06 (m, 1H).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 175.75, 173.60, 171.31, 136.81, 135.18, 133.72, 131.58, 130.81, 129.70, 129.18, 127.66, 67.52, 61.99, 53.95, 49.36, 45.09, 37.40, 24.19, 21.66

Mass spectroscopy: (+ FAB) 447 (M + H).

Example 20

Synthesis of N- (toluene-4-sulfonyl) -L-thiaprolyl-L-phenylalanine

N- (toluene-4-sulfonyl) -L-thiazolidine-4-carboxylic acid is prepared from L-thiazolidine-4-carboxylic acid using the procedure described in Method 1. Using the procedure described in Example 1 (19), the title compound having a melting point of 60-65 ° C. is prepared as a solid.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 2.43 (s, 5H), 3.0-3.40 (m, 3H), 4.0 (d, 1H, J = 15.7 Hz), 4.55 (d, 1H, J = 15.8 Hz ), 4.64 (m, 1H), 4.87 (m, 1H), 7.19-7.41 (m, 7H), 7.72 (m, 2H).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 22.3, 33.6, 37.7, 52.0, 53.0, 65.8, 127.8, 128.5, 129.4, 129.9, 130.7, 133.9, 136.2, 145.7, 169.4, 175.0.

Mass spectroscopy: (FAB +) 435 (M + H).

Example 21

Synthesis of N- (3,5-dichlorobenzenesulfonyl) -L-propyl-L-phenylalanine

Proline methyl ester hydrochloride (2.68 g, 16.2 mmol) is dissolved in pyridine (20 mL), 3,5-dichlorophenylsulfonyl chloride (3.57 g, 14.6 mmol) is added to the mixture and then stirred for 19 h. Water (5 mL) is added and stirred for 45 minutes combined before dilution with water (200 mL). The mixture is extracted with Et ₂ O (2 × 150 mL), the combined extracts are washed with water (3 × 100 mL), 1N HCl (150 mL) and saturated aqueous NaHCO ₃ (150 mL), then dried (MgSO ₄ ), Filtration and distillation under vacuum gave 3,5-dichlorophenylsulfonylproline methyl ester (4.34 g, 88%) as an oil. Methyl ester is dissolved in MeOH (20 mL) and 1N NaOH (20 mL) is added. Warm gently until the mixture is homogeneous and stir for 1.5 hours. The solvent is removed in vacuo and the residue is worked up in water (50 mL). The aqueous solution is washed with Et ₂ O (30 mL) and acidified with 12N HCl. The mixture is extracted with CHCl ₃ (2 × 40 mL), the combined extracts are dried over anhydrous MgSO ₄ , filtered and the solvent is distilled under vacuum to give 3,5-dichlorophenylsulfonyl-L-proline (3.37 g , 83%) is obtained as a solid. 3,5-dichlorophenylsulfonyl-L-proline was coupled with L-phenylalanine ethyl ester using the procedure described in Method 3 above to give the corresponding dipeptide ethyl ester (348 mg, 45%). The title compound is prepared via hydrolysis of ethyl ester with NaOH in MeOH (61 mg, 19%). NMR analysis indicates the presence of diastereomers.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.11 (bs, 1H), 7.72 (d, 2H, J = 1.9 Hz), 7.61 (q, 1H, J = 2.0 Hz), 7.39-7.20 (6H), 4.92 ( m, 1H), 4.15 (m, 1H), 3.52-3.09 (4H), 2.11 (m, 1H), 1.95-1.49 (3H).

¹³ C NMR (CDCl ₃ ): δ = 175.4, 175.2, 171.8, 171.7, 139.5, 139.3, 137.0, 136.6, 135.8, 134.0, 134.0, 129.9, 129.5, 127.7, 126.7, 111.4, 63.1, 62.9, 53.8, 53.6, 50.5, 50.3, 45.1, 37.9, 31.3, 30.8, 24.8, 24.6.

Mass spectroscopy: FAB m / e 471 (M + H).

Example 22

Synthesis of N- (4-trifluoromethoxybenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared as described in Example 21 (59), except that 3,5-dichlorophenylsulfonyl chloride is used instead of 4-trifluoromethoxyphenylsulfonyl chloride.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 7.90 (d, 2H, J = 8.8 Hz), 7.38-7.21 (8H), 6.90 (bs, 1H), 4.90 (m, 1H), 4.15 (dd, 1H, J = 2.5, 8.4 Hz), 3.38 (m, 2H), 3.12 (m, 2H), 1.98 (m, 1H), 1.55 (2H), 1.41 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 174.9, 172.0, 153.3, 136.6, 134.9, 130.6, 129.9, 129.2, 127.7, 121.7, 62.8, 53.8, 50.2, 37.9, 30.6, 24.6.

Mass spectroscopy: FAB m / e 487 (M + H).

Example 23

Synthesis of N- (3,4-dichlorobenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared as described in Example 21 (59), except that 3,5-dichlorophenylsulfonyl chloride is used instead of 3,4-dichlorophenylsulfonyl chloride.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 7.94 (d, 1H, J = 1.9 Hz), 7.64 (m, 2H), 7.55 (bs, 1H), 7.36-7.21 (6H), 4.92 (dt, 1H, J = 5.2, 8.0 Hz), 4.16 (m, 1H), 3.36 (m, 2H), 3.11 (m, 1H), 1.98 (m, 1H), 1.58 (m, 2H), 1.40 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 175.0, 171.8, 139.5, 136.4, 134.7, 132.1, 130.2, 129.9, 129.2, 127.7, 127.4, 62.9, 53.8, 50.3, 37.9, 30.7, 24.6.

Mass spectroscopy: FAB m / e 471 (M + H).

Example 24

Synthesis of N- (toluene-4-sulfonyl) -D, L- (trans-3-phenyl) prolyl-L-phenylalanine

N- (toluene-4-sulfonyl) -trans-3-phenylproline was described using the procedure described in Method 3. See Chung et al., J. Org. Chem., 55: 270-275 (1990)}, was coupled with L-phenylalanine ethyl ester and flash chromatographed with silica gel (93/7 CH ₂ Cl ₂ / MeOH) to give the title compound. Ethyl ester is obtained as a white solid. The acid is prepared by hydrolyzing the ethyl ester with NaOH in ethanol.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ ): δ = 8.38 (m, 1H), 7.70 (m, 2H), 7.40-7.06 (10H), 6.80 (m, 1H), 6.62 (m, 1H), 4.50 (m , 1H), 4.13 (d, 0.5H, J = 5.2 Hz), 3.97 (d, 0.5H, J = 5.5 Hz), 3.46 (m, 2H), 3.02 (m, 3H), 2.43 (s, 1 / 2 X 3H), 2.42 (s, 1/2 X 3H), 2.07 (m, 1H), 1.45 (m, 1H).

¹³ C NMR (DMSO-d ₆ ): δ = 172.9, 172.8, 170.7, 170.7, 144.1, 143.9, 141.4, 141.3, 138.0, 137.7, 134.9, 134.6, 130.2, 130.1, 129.8, 129.6, 128.7, 128.6, 128.5, 127.8, 127.7, 127.2, 127.1, 126.9, 126.8, 126.7, 79.6, 67.9, 53.6, 53.5, 49.7, 49.5, 49.5, 49.1, 37.2, 37.1, 32.5, 32.3, 21.4.

Mass spectroscopy: FAB m / e 493 (M + H).

Example 25

Synthesis of N- (3,4-dimethoxybenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared as described in Example 21 (59), except that 3,5-dichlorophenylsulfonyl chloride is used instead of 3,4-dimethoxyphenylsulfonyl chloride. NMR analysis with epimerization showed a diastereomeric mixture of about 65:35.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 7.46 (m, 1H), 7.39-7.20 (6H), 6.97 (dd, 1H, J = 4.5, 8.4 Hz), 4.85 (m, 1H), 4.10 (m, 1H ), 3.96 (s, 0.35 X 3H), 3.95 (s, 0.65 X 3H), 3.93 (s, 0.35 X 3H), 3.92 (s, 0.65 X 3H), 3.44-3.22 (2H), 3.11 (m, 2H) ), 2.01 (m, 1 H), 1.60-1.33 (3 H).

¹³ C NMR (CDCl ₃ ): δ = 175.1, 174.7, 172.5, 172.3, 153.7, 149.8, 136.6, 135.9, 130.2, 129.9, 129.4, 129.1, 128.0, 128.0, 127.9, 127.7, 122.4, 111.4, 110.7, 63.2, 62.8, 57.0, 56.8, 53.8, 53.6, 50.3, 50.2, 38.0, 31.1, 30.5, 24.8, 24.7.

Mass spectroscopy: FAB m / e 463 (M + H).

Example 26

Synthesis of N- (benzene-4-sulfonyl) -D, L- (cis-3-phenyl) prolyl-L-phenylalanine

N-acetyl-cis-3-phenylproline ethyl ester (614 mg, 2.35 mmol) [Chung et al., J. Org. Chem., 55: 270-275 (1990)} is dissolved in HOAc (4 mL) and 6N HCl (12 mL) and heated at reflux for 18 h. The mixture is cooled to room temperature and the volatiles are distilled under vacuum to give a white solid. Dissolve the solid in 1N NaOH (15 mL), add dioxane (10 mL), then add 4-toluenesulfonyl chloride (458 mg, 2.40 mmol). The mixture is stirred at room temperature for 18 hours before 6N HCl is added to bring the pH below 4. Extract the mixture with Et ₂ O (3 × 40 mL), dry the extract (MgSO ₄ ), filter and distill under vacuum to give N- (toluene-4-sulfonyl) -cis-3-phenylproline (765 mg). , 94%) is obtained as a white solid.

The N- (toluene-4-sulfonyl) -cis-3-phenylproline is coupled with phenylalanine ethyl ester using the procedure described in Method 3. The title compound is prepared by hydrolysis of ethyl ester using NaOH in ethanol.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.57 (bs, 1H), 7.77 (m, 2H), 7.31-6.70 (12H), 4.65 (m, 0.5H), 4.47 (m, 0.5H), 4.41 (d , 0.5H, J = 8.9Hz, 4.34 (d, 0.5H, J = 9.0Hz), 3.73 (m, 1H), 3.28-2.76 (4H), 2.44 (s, 3H), 2.40 (m, 0.5H ), 2.17 (m, 0.5H), 1.8 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 174.5, 173.9, 169.5, 168.8, 144.3, 136.1, 135.8, 135.6, 135.3, 133.4, 133.1, 129.9, 129.6, 129.4, 128.6, 128.6, 128.3, 127.8, 127.7, 127.5, 127.1, 127.0, 65.9, 65.8, 53.6, 52.5, 48.5, 48.3, 48.2, 37.5, 37.3, 39.0, 28.6, 21.6.

Mass spectroscopy: FAB m / e 493 (M + H).

Example 27

Synthesis of N- (4-nitrobenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound (melting point 70 to 80 ° C.) was prepared as described in Example 21 (59), except that 3,5-dichlorophenylsulfonyl chloride was used instead of 4-nitrophenylsulfonyl chloride.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.38 (d, 2H, J = 8.9 Hz), 8.02 (d, 2H, J = 8.9 Hz), 7.32-7.20 (6H), 4.90 (m, 1H), 4.25 ( dd, 1H, J = 2.5, 8.4 Hz), 3.42 (m, 2H), 3.16 (m, 2H), 1.98 (m, 1H), 1.62 (m, 2H), 1.54 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 175.0, 171.8, 151.1, 142.5, 136.4, 129.9, 129.7, 127.8, 125.2, 62.9, 53.7, 50.3, 37.9, 30.9, 24.7.

Mass spectroscopy: FAB m / e 448 (M + H).

Example 28

Synthesis of N- (4-acetamidobenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared as described in Example 21 (59), except that 3,5-dichlorophenylsulfonyl chloride is used instead of 4-acetamido-phenylsulfonyl chloride.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ ): δ = 12.85 (bs, 1H), 10.4 (bs, 1H), 8.07 (d, 1H, J = 8.2 Hz), 7.76 (ab q, 4H), 7.25 (m, 5H), 4.49 (dd, 2H, J = 3.2, 8.2 Hz), 4.06 (m, 1H), 3.36 (m, 1H), 3.08 (3H), 2.09 (s, 3H), 1.58-1.48 (4H).

¹³ C NMR (DMSO-d ₆ ): δ = 173.0, 171.2, 169.5, 143.9, 137.8, 130.4, 129.7, 129.1, 128.5, 126.8, 119.0, 61.7, 53.5, 49.4, 37.0, 30.7, 24.6, 24.1.

Mass spectroscopy: FAB m / e 460 (M + H).

Example 29

Synthesis of N- (4-cyanobenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared as described in Example 21 (59), except that 3,5-dichlorophenylsulfonyl chloride is used instead of 4-cyanophenyl-phenylsulfonyl chloride.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.40 (bs, 1H), 7.95 (d, 2H, J = 8.3 Hz), 8.07 (d, 2H, J = 8.3 Hz), 7.36-7.20 (6H), 4.93 ( dt, 1H, J = 5.4, 8.0 Hz), 4.22 (m, 1H), 3.38 (m, 2H), 3.13 (m, 2H), 1.92 (m, 1H), 1.56 (m, 2H), 1.40 (m , 1H).

¹³ C NMR (CDCl ₃ ): δ = 175.0, 171.9, 140.8, 136.5, 133.8, 129.9, 129.2, 129.0, 127.8, 117.8, 117.7, 62.8, 53.7, 50.3, 37.9, 30.9, 24.6.

Mass spectroscopy: FAB m / e 428 (M + H).

Example 30

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-tryptophan

The N- (toluene-4-sulfonyl) -L-proline hydrate is coupled with L-tryptophan using the procedure described in Method 3. The title compound is prepared by hydrolysis of the methyl ester with LiOH in THF / water.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.45 (d, 1H, J = 2.0 Hz), 7.64 (d, 2H, J = 8.2 Hz), 7.60 (d, 1H, J = 7.6 Hz), 7.45 (d, 1H, J = 7.4 Hz), 7.33 (d, 1H, J = 8.0 Hz), 7.27 (d, 2H, J = 8.0 Hz), 7.19-7.06 (3H), 4.88 (m, 1H), 4.07 (dd, 1H, J = 2.9, 8.6Hz), 3.48 (dd, 1H, J = 5.4, 14.8Hz), 3.36 (dd, 1H, J = 7.1, 14.8Hz), 3.03 (m, 2H), 2.40 (s, 3H ), 1.88 (m, 1H), 1.47 (m, 1H), 1.32 (m, 1H), 1.22 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 174.8, 172.7, 145.0, 136.7, 133.3, 130.6, 128.4, 128.3, 124.3, 122.6, 118.9, 112.0, 110.0, 62.8, 54.0, 50.1, 30.5, 27.5, 24.6, 22.1.

Mass spectroscopy: FAB m / e 456 (M + H).

Example 31

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-β- (1-naphthyl) -L-alanine

The N- (toluene-4-sulfonyl) -L-proline hydrate is coupled with β- (1-naphthyl) alanine methyl ester hydrochloride using the procedure described in Method 3. The title compound is prepared by hydrolysis of the methyl ester with LiOH in THF / water.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.16 (d, 1H, J = 8.2 Hz), 8.05 (bs, 1H), 7.85 (d, 1H, J = 6.9 Hz), 7.77-7.27 (10H), 4.99 ( m, 1H), 4.05 (m, 1H), 3.90 (dd, 1H, J = 5.0, 14.3 Hz), 3.51 (dd, 1H, J = 9.6, 14.3 Hz), 3.11 (m, 1H), 3.02 (m , 1H), 2.41 (s, 3H), 1.74 (m, 1H), 1.34 (m, 2H), 1.00 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 174.9, 172.5, 145.0, 134.4, 133.4, 133.1, 132.6, 130.6, 129.4, 128.5, 128.4, 127.2, 126.4, 126.0, 14.0, 62.7, 53.9, 50.1, 34.9, 30.3, 24.5, 22.1.

Mass spectroscopy: FAB m / e 467 (M + H).

Example 32

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-β- (2-naphthyl) -L-alanine

The N- (toluene-4-sulfonyl) -L-proline hydrate is coupled with β- (2-naphthyl) -L-alanine methyl ester hydrochloride using the procedure described in Method 3. The title compound is prepared by hydrolysis of the methyl ester with LiOH in THF / water.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.20 (bs, 1H), 7.80-7.67 (6H), 7.55 (d, 1H, J = 7.9 Hz), 7.46-7.27 (5H), 4.99 (m, 1H), 4.12 (m, 1H), 3.55 (m, 1H), 3.27 (dd, 1H, J = 8.2, 14.1 Hz), 3.17 (m, 1H), 3.01 (m, 1H), 2.39 (s, 3H), 1.83 (m, 1 H), 1.40-1.26 (3 H).

¹³ C NMR (CDCl ₃ ): δ = 174.8, 172.4, 145.0, 134.3, 133.9, 133.4, 133.0, 130.6, 128.9, 128.8, 128.4, 128.2, 128.1, 127.9, 126.7, 126.3, 62.7, 53.9, 50.1, 38.2, 30.4, 24.5, 22.1.

Mass spectroscopy: FAB m / e 467 (M + H).

Example 33

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-β- (2-thienyl) -L-alanine

The N- (toluene-4-sulfonyl) -L-proline hydrate is coupled with β- (2-thienyl) -L-alanine methyl ester hydrochloride using the procedure described in Method 3. The title compound is prepared by hydrolysis of the methyl ester with LiOH in THF / water.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.42 (bs, 1H), 7.73 (d, 2H, J = 8.3 Hz), 7.57 (d, 1H, J = 7.4 Hz), 7.34 (d, 2H, J = 7.9 Hz), 7.16 (dd, 1H, J = 1.2, 5.1 Hz), 6.95-6.88 (m, 2H), 4.87 (m, 1H), 4.18 (m, 1H), 3.56-3.41 (3H), 3.17 ( m, 1H), 2.43 (s, 3H), 2.08 (m, 1H), 1.60 (m, 3H).

¹³ C NMR (CDCl ₃ ): δ = 174.2, 172.6, 145.1, 137.9, 133.4, 130.6, 128.5, 127.7, 127.6, 125.4, 62.8, 54.0, 50.4, 32.11, 30.7, 24.9, 22.2.

Mass spectroscopy: FAB m / e 423 (M + H).

Example 34

Synthesis of N- (isopropanesulfonyl) -L-prolyl-L-phenylalanine

CH ₃ SO ₂ Cl is substituted with (CH ₃ ) ₂ CHSO ₂ Cl and the title compound is obtained as a clear oil by the procedure described in Example 1 (9).

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.10 (d, J = 7.9, 1 Hz), 7.28-7.16 (m, 5H), 4.48-4.41 (m, 1H), 4.27-4.24 (m, 1H ), 3.47-3.23 (m, 3H), 3.11-3.00 (m, 2H), 2.89 (dd, J = 13.9, J = 9.4, 1H), 2.11-2.01 (m, 1H), 1.82-1.71 (m, 3H), 1.11 (d, J = 6.8, 6H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.6, 171.1, 137.4, 129.2, 128.1, 126.4, 61.2, 53.0, 48.6, 43.6, 36.5, 30.6, 24.0, 20.0, 19.8.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 369 (MH &lt; + &gt;).

Example 35

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-β- (3-pyridyl) -L-alanine

N- (toluene-4-sulfonyl) -L-proline hydrate was coupled with L-β- (3-pyridyl) alanine methyl ester dihydrochloride using the procedure described in Method 3 to produce N- (toluene-4 -Sulfonyl) prolyl-L-β- (3-pyridyl) alanine is obtained. The title compound is prepared by hydrolysis of the methyl ester using 0.5N aqueous NaOH in THF / water.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ ): δ = 8.33 (dd, 1H, J = 1.4, 4.7 Hz), 8.27 (d, 1H, J = 1.9 Hz), 7.75 (d, 2H, J = 8.2 Hz), 7.71 (m, 1H), 7.52 (m, 1H), 7.42 (d, 2H, J = 8.5 Hz), 7.19 (dd, 1H, J = 4.7, 7.7 Hz), 4.00 (m, 2H), 3.20-3.00 (4H), 2.40 (s, 3H), 1.72 (m, 1H), 1.40 (3H).

¹³ C NMR (DMSO-d ₆ ): δ = 172.7, 170.0, 150.9, 147.4, 144.1, 137.4, 134.6, 133.8, 130.3, 128.1, 123.1, 62.4, 55.1, 49.3, 34.4, 30.5, 24.00, 21.4.

Mass spectroscopy: FAB m / e 440 (M + Na).

Example 36

Synthesis of N- (toluene-4-sulfonyl) -L- (cis-4-phenylthio) prolyl-L-phenylalanine

Trans-4-hydroxy-L-proline is treated with EtOH and HCl gas and the mixture is distilled to give trans-4-hydroxy-L-proline ethyl ester hydrochloride. This product was treated with TsCl in pyridine, aqueous workup, and then N- (toluene-4-sulfonyl) -trans-4- (O- (toluene-4-sulfonyl))-L-proline ethyl ester To obtain. This product was treated with PhSH and DBU in DMF, aqueous workup and flash chromatography on silica gel to give N- (toluene-4-sulfonyl) -cis-4- (phenylthio) -L-proline ethyl ester do. This product was treated with NaOH in CH ₃ OH and H ₂ O, oxidized, extracted, dried over MgSO ₄ and then N- (toluene-4-sulfonyl) -cis-4- (phenylthio) -L-proline To obtain. This product was treated with HCl.Phe-OMe, BOP and NMM in DMF, post-treated and flash chromatographed, followed by N- (toluene-4-sulfonyl) -cis-4- (phenylthio) -Pro-Phe -OMe is obtained. This product is treated with NaOH in CH ₃ OH and H ₂ O, oxidized, extracted, dried over MgSO ₄ , filtered and evaporated to afford the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.19 (d, J = 8.0, 1H), 7.71 (d, J = 8.3, 2H), 7.41 (d, J = 8.2, 2H), 7.35-7.14 (m, 10H), 4.53-4.46 (m, 1H), 4.24 (t, J = 7.8, 1H), 3.73 (dd, J = 11.6, J = 6.7, 1H), 3.36 (bs, 1H), 3.17- 2.86 (m, 4H), 2.39 (s, 3H), 2.34-2.25 (m, 1H), 1.57-1.47 (m, 1H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.4, 170.0, 144.0, 137.3, 134.1, 133.9, 130.0, 129.8, 129.4, 129.3, 128.2, 127.5, 126.9, 126.5, 61.0, 54.9, 53.2, 42.3, 36.7, 36.4, 21.1.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 525 (MH &lt; + &gt;).

Example 37

Synthesis of N- (toluene-4-sulfonyl) -L- (cis-4-benzylthio) prolyl-L-phenylalanine

Phenylthiol is substituted with benzinethiol and the title compound is obtained as a pure oil by the procedure described in Example 36 (131).

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.18 (d, J = 8.1, 1H), 7.65 (d, J = 8.3, 2H), 7.38 (d, J = 8.2, 2H), 7.29-7.17 (m, 10H), 4.50-4.43 (m, 1H), 4.05 (t, J = 7.9, 1H), 3.74-3.56 (m, 3H), 3.35 (bs, 1H), 3.09-2.94 (m, 4H) , 2.40 (s, 3H), 2.20-2.10 (m, 1H), 1.40-1.28 (m, 1H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.4, 170.2, 143.8, 138.5, 137.3, 134.0, 130.0, 129.4, 128.7, 128.4, 128.2, 127.3, 126.9, 126.5, 60.9, 55.5, 53.1, 39.3, 36.7, 36.6, 34.9, 21.1.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 539 (MH &lt; + &gt;).

Example 38

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-histidine

N- (toluene-4-sulfonyl) prolyl-L- (N-benzyl) histidine (237 mg, 0.464 mmol) is dissolved in MeOH (20 mL) and 10% Pd / C (50 mg) is added. The mixture is hydrogenated under H ₂ 50 psi for 36 h. The mixture is filtered to remove the catalyst and the filtrate is distilled under vacuum. The residue was purified by preparative TLC (90: 10: 1 CH ₂ Cl ₂ / MeOH / NH ₄ OH) to give N- (toluene-4-sulfonyl) -L-prolyl-L-histidine methyl ester (155 mg). , 79%). The title compound (128 mg, 81%) is prepared by hydrolysis of the methyl ester with 0.5N NaOH in THF / water.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ ): δ = 7.77 (d, 3H, J = 6.5 Hz), 7.42 (m, 3H), 4.03 (m, 1H), 3.99 (m, 1H), 3.22 (m, 1H ), 3.05 (m, 2H), 2.95 (m, 1H), 2.41 (s, 3H), 1.80 (m, 1H), 1.45 (3H).

¹³ C NMR (DMSO-d ₆ ): δ = 174.0, 169.8, 144.1, 133.8, 10.3, 128.0, 62.4, 49.4, 40.0, 30.6, 24.0, 21.4.

Mass spectroscopy: FAB m / e 429 (M + Na).

Example 39

Synthesis of N- (toluene-4-sulfonyl) -L- (cis-4-amino) prolyl-L-phenylalanine

Trans-4-hydroxy-L-proline is treated with EtOH and HCl gas and the mixture is distilled to give trans-4-hydroxy-L-proline ethyl ester hydrochloride. This product is treated with TsCl in pyridine and after aqueous workup to afford N- (toluene-4-sulfonyl) -trans-4- (TsO) -L-proline ethyl ester. This product is treated with NaN ₃ in DMF and H ₂ O, aqueous workup and flash chromatography to yield N- (toluene-4-sulfonyl) -cis-4-azido-L-proline ethyl ester . This product was treated with NaOH in CH ₃ OH and H ₂ O, oxidized, extracted and dried with MgSO ₄ to afford N- (toluene-4-sulfonyl) -cis-4-azido-L-proline do. This product is treated with 10% Pd / C in THF, AcOH and H ₂ O and the mixture is stirred under 50 psi of H ₂ . The mixture is filtered through celite and distilled to afford N- (toluene-4-sulfonyl) -cis-4-amino-L-proline. This product was treated with Boc ₂ O and Et ₃ N in t-BuOH and H ₂ O, distilled, oxidized, extracted, dried over MgSO ₄ , filtered and evaporated to give N- (toluene-4-sulfonyl)- Cis-4- (t-butoxycarbonylamino) -L-proline is obtained. This product was treated with HCl.Phe-Ot-Bu, BOP and NMM in DMF, followed by aqueous workup and flash chromatography, followed by N- (toluene-4-sulfonyl) -cis-4- (t-butoxy Carbonylamino) -Pro-Phe-Ot-Bu is obtained. This product is treated with TFA and distilled to afford the title compound trifluoroacetate salt as a clean oil.

NMR data are as follows:

¹ H NMR (CD ₃ OD, 300 MHz): δ = 7.47 (d, J = 8.3, 2H), 7.22-7.00 (m, 7H), 4.47 (dd, J = 7.9, J = 5.3, 1H), 4.18 ( dd, J = 9.8, J = 1.7, 1H), 3.64 (t, J = 5.2, 1H), 3.43 (d, J = 11.2, 1H), 3.22 (dd, J = 11.3, J = 5.0, 1H), 3.01 (dd, J = 14.1, J = 5.3, 1H), 2.88 (dd, J = 14.0, J = 7.9, 1H), 2.23 (s, 3H), 2.21-2.08 (m, 1H), 1.87-1.80 ( m, 1 H).

¹³ C NMR (CD ₃ OD, 75 MHz): δ = 174.7, 173.9, 146.1, 138.0, 134.7, 131.1, 130.5, 129.6, 128.7, 128.0, 60.7, 55.6, 53.9, 51.5, 38.2, 35.3, 21.5.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 432 (MH &lt; + &gt;).

Example 40

Synthesis of N- (toluene-4-sulfonyl) -L- (trans-4-amino) prolyl-L-phenylalanine

Trans-4-hydroxy-L-proline was substituted with cis-4-hydroxy-L-proline and the preparation of Example 39 (136) gave the title compound trifluoroacetate salt as a clean oil. .

NMR data are as follows:

¹ H NMR (CD ₃ OD, 300 MHz): δ = 7.39 (d, J = 8.4, 2H), 7.13-7.01 (m, 7H), 4.40-4.33 (m, 1H), 4.28 (dd, J = 9.0, J = 2.2, 1H), 3.59-3.50 (m, 2H), 3.07-2.97 (m, 2H), 2.81 (dd, J = 14.0, J = 8.9, 1H), 2.21 (s, 3H), 1.24-2.06 (m, 1 H), 1.80-1.68 (m, 1 H).

¹³ C NMR (CD ₃ OD, 75 MHz): δ = 174.1, 172.6, 145.9, 138.3, 135.0, 131.0, 130.5, 129.6, 128.9, 128.0, 61.1, 55.2, 52.0, 49.7, 38.2, 35.4, 21.5.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 432 (MH &lt; + &gt;).

Example 41

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine benzyl ester

The N- (toluene-4-sulfonyl) -L-proline is coupled with the L-phenylalanine bezel ester toluenesulfonic acid salt using the procedure described in Method 3 to afford the title compound as an oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 7.68 (d, 2H, J = 8.2 Hz), 7.39-7.17 (10H), 7.05 (m, 1H), 5.22 (d, 2H, J = 12.2), 5.14 (d , 1H, J = 12.2Hz), 4.88 (m, 1H), 4.08 (m, 1H), 3.28 (m, 3H), 3.08 (m, 2H), 2.39 (s, 3H), 2.00 (m, 1H) , 1.46 (m, 3 H).

¹³ C NMR (CDCl ₃ ): δ = 171.4, 171.3, 144.9, 136.5, 135.8, 133.6, 130.6, 129.9, 129.1, 129.1, 129.0, 129.0, 128.4, 127.6, 67.8, 62.8, 54.0, 50.2, 38.4, 30.4, 24.8, 22.2.

Mass spectroscopy: FAB m / e 507 (M + H).

Example 42

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-methoxyamide

Mixed anhydride processes [Greenstein, J. P. and Milton Wenitz, "Chemistry of the Amino Acids", Volume 2, pp. 978-979, Robert E. Krieger Publishing Co., Malabar, FL. (1961)]. Coupling with chloride affords the title compound as an oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 9.66 (s, 1H), 7.70 (d, 2H, J = 8.2 Hz), 7.36 (d, 2H, J = 8.0 Hz), 7.33-7.21 (5H), 6.90 ( d, 1H, J = 9.5 Hz), 4.88 (m, 1H), 3.81 (m, 1H), 3.75 (s, 3H), 3.46 (m, 2H), 3.07 (m, 2H), 2.45 (s, 3H ), 1.60 (m, 2H), 1.47 (m, 2H).

¹³ C NMR (CDCl ₃ ): δ = 171.7, 168.1, 145.6, 137.0, 131.8, 130.7, 129.7, 129.3, 128.5, 127.7, 64.9, 62.7, 52.6, 50.6, 37.6, 31.4, 24.7, 22.2.

Mass spectroscopy: FAB m / e 446 (M + H).

Example 43

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-benzyloxyamide

The N- (toluene-4-sulfonyl) -L-proline-L-phenylalanine was coupled with O-benzylhydroxyamine using the procedure described in Method 3 to give the title compound having a melting point of 127 to 130 ° C. as a white solid. To obtain.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 9.54 (s, 1H), 7.61 (d, 2H, J = 8.2 Hz), 7.43-7.19 (12H), 6.86 (d, 1H, J = 9.3 Hz), 4.91 ( m, 2H), 4.82 (m, 1H), 3.72 (m, 1H), 3.42 (m, 2H), 3.07 (m, 2H), 2.45 (s, 3H), 1.64 (m, 2H), 1.44 (m , 1H), 1.26 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 171.6, 168.2, 145.5, 136.9, 135.7, 132.0, 130.7, 129.9, 129.7, 129.0, 128.9, 128.5, 127.7, 78.8, 62.7, 52.7, 50.6, 37.6, 31.2, 24.7, 22.2.

Mass spectroscopy: FAB m / e 522 (M + H).

Example 44

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N- (toluene-4-sulfonyl) amide

N- (toluene-4-sulfonyl) -L-proline is reacted with L-phenylalanine amide using the procedure described in Method 12 to N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine amide To obtain. Add N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine amide to a solution of NaH (60% mineral oil prewashed with THF) at TH ° C. at 0 ° C. and react for 45 minutes at 0 ° C. Stir. Toluene-4-sulfonyl chloride is added and the reaction is stirred for 16 hours at room temperature. The reaction mixture was extracted with EtOAc (3 × 50 mL) and 0.2N HCl (50 mL), the combined organic layers were washed successively with saturated NaHCO ₃ (50 mL) and saturated NaCl (2 × 50 mL) and dried over MgSO ₄ . , Filtration and concentration to afford the title compound as an oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.97 (d, 2H, 8.52 Hz), 7.72 (d, 2H, 8.52 Hz), 7.37-7.04 (m, 10H), 4.80 (m, 1H), 3.86 ( m, 1H), 3.42 (m, 1H), 3.29 (m, 1H), 3.01 (m, 2H), 2.45 (bs, 6H), 1.65 (m, 2H), 1.45 & 1.31 (m, 2H).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 172.46, 169.93, 145.47, 136.43, 132.06, 130.70, 130.07, 129.58, 129.32, 129.17, 128.61, 127.74, 62.68, 54.52, 50.57, 36.91, 31.10, 24.69, 22.69, 22. 22.23.

Mass spectroscopy: (+ FAB) 570 (M + H).

Preliminary Example A

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-D, L-phenylalanyl-β-alanine ethyl ester

BOP (about 2.1 equivalents) and NMM (about 4 equivalents) are added to N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine (about 2 equivalents) in DMF and the reaction is about 45 at room temperature. Stir for minutes. β-alanine ethyl ester (about 2 equivalents) is added and the reaction is stirred at room temperature for about 16 hours. The reaction mixture is extracted with water and diethyl ether. The combined organic solution is washed with 0.2N HCl, saturated NaHCO ₃ and saturated NaCl solution. The organic layer is dried over anhydrous magnesium sulfate and concentrated by filtration. The residue is purified by silica gel chromatography (50% EtOAc / hexanes, R _f = 0.18) to afford the title compound, measured as a colorless oil as a mixture of diastereomers by NMR.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.68 (m, 2H), 7.25 (m, 9H), 4.07 (m, 3H), 3.45 (m, 4H), 3.06 (m, 3H), 2.50 & 2.34 (m, 3H), 2.41 (s, 3H), 1.62 (m, 4H), 1.20 (t, 3H, J = 5.55 Hz).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 177.62, 172.72, 171.98, 171.73, 171.25, 170.80, 145.39, 145.02, 137.52, 137.20, 133.28, 132.09, 133.28, 132.09, 130.66, 130.57, 130.02, 129 1859 129.49, 128.44, 127.49, 127.46, 63.06, 62.75, 61.18, 61.12, 55.49, 54.17, 50.62, 50.36, 38.27, 37.92, 35.98, 35.32, 34.47, 34.30, 31.27, 30.95, 24.92, 24.62, 22.18, 14.72.

Mass spectroscopy: (+ FAB) 516 (M + H).

Example 45

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-D, L-phenylalanyl-β-alanine

From Preliminary Example A 161 using the procedure described in Example 19 (57), the title compound as a translucent solid was prepared, which determined that the melting point was 91-95 ° C. and was a mixture of diastereomers by NMR.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.70 & 7.27 (m, 9H), 4.83 (m, 0.5H), 4.05 (m, 0.5H), 3.47 (m, 4H), 3.06 (m, 3H) , 2.55 (m, 2H), 2.43 (s, 3H), 1.62 (m, 4H).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 175.85, 175.68, 172.45, 172.23, 171.67, 171.43, 145.32, 144.96, 137.37, 136.91, 133.42, 132.37, 130.67, 130.56, 130.52, 130.03, 129.12, 129.12. 128.51, 128.48, 127.52, 63.01, 62.72, 55.25, 54.34, 50.52, 50.43, 50.39, 38.81, 38.07, 35.82, 35.79, 35.25, 35.23, 34.17, 34.10, 34.08, 31.23, 24.91, 24.61, 22.18.

Mass spectroscopy: (+ FAB) 488 (M + H).

Example 46

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-hydroxyamide

N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-benzyloxyamide [see Example 43 (159)] was dissolved in MeOH (1 mL) and 5% Pd / BaSO ₄ (52 mg) Add). The mixture is hydrogenated at H ₂ 50 psi for 9 hours. The mixture was filtered through a pad of diatomaceous earth and distilled under vacuum to afford a residue which was purified by silica gel chromatography (92: 8 CH ₂ Cl ₂ / MeOH) to afford the title compound as an oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 9.85 (br s, 1H), 7.73 (d, 2H, J = 7.2 Hz), 7.36 (d, 2H, J = 7.0 Hz), 7.28-7.20 (5H), 7.07 (m, 1H), 4.96 (m, 1H), 3.87 (m, 1H), 3.47 (m, 2H), 3.07 (m, 2H), 2.45 (s, 3H), 1.90 (br s, 1H), 1.63 (m, 2H), 1.46 (m, 1H), 1.36 (m, 1H).

¹³ C NMR (CDCl ₃ ): δ = 171.8, 168.4, 145.5, 136.9, 132.0, 130.7, 129.7, 129.2, 128.6, 127.6, 62.7, 52.2, 50.6, 38.0, 31.2, 24.6, 22.2.

Mass spectroscopy: FAB m / e 432 (M + H).

Example 47

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine isopropyl ester

The N- (toluene-4-sulfonyl) -L-proline hydrate is coupled with phenylalanine isopropyl ester trifluoroacetate using the procedure described in Method 3 to afford the title compound as an oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 7.68 (d, 2H, J = 8.2 Hz), 7.33-7.12 (8H), 5.05 (p, 1H, J = 6.3 Hz), 4.77 (m, 1H), 4.06 ( m, 1H), 3.33 (m, 1H), 3.22 (dd, 1H, J = 5.8, 14.0 Hz), 3.06 (m, 2H), 2.40 (s, 3H), 1.99 (m, 1H), 1.46 (3H ), 1.23 (d, 6H, J = 6.3 Hz).

¹³ C NMR (CDCl ₃ ): δ = 171.3, 170.9, 144.9, 136.7, 133.5, 130.5, 129.9, 129.0, 128.4, 127.5, 70.0, 62.9, 54.0, 50.2, 38.5, 30.4, 24.7, 22.3, 22.2, 22.1.

Mass spectroscopy: FAB m / e 459 (M + H).

Example 48

Synthesis of N- (toluene-4-sulfonyl) -L- (cis-4-hydroxy) -propyl-L-phenylalanine

In the preparation of Example 6 (17), D-Pro-OH was substituted with cis-4-hydroxy-L-proline and the title compound was obtained as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.14 (d, J = 7.9, 1H), 7.70 (d, J = 8.2, 2H), 7.41 (d, J = 8.2, 2H), 7.28-7.15 (m, 5H), 5.30 (bs, 1H), 4.52-2.45 (m, 1H), 4.15 (dd, J = 9.0, J = 4.1, 1H), 3.85 (bs, 1H), 3.31 (bs, 1H) , 3.24 (dd, J = 10.6, J = 4.9, 1H), 3.17 (dd, J = 10.9, J = 3.4, 1H), 3.01-2.94 (m, 2H), 2.40 (s, 3H), 1.92-1.82 (m, 1 H), 1.79-1.69 (m, 1 H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.2, 171.2, 143.8, 136.9, 133.4, 129.9, 129.5, 128.1, 127.5, 126.5, 68.6, 60.3, 56.4, 53.4, 38.0, 37.0, 21.0.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 433 (MH &lt; + &gt;).

Example 49

Synthesis of N- (toluene-4-sulfonyl) -L- (trans-4-fluoro) -prolyl-L-phenylalanine benzyl ester

In the preparation of Example 50 (177), trans-4-hydroxy-L-proline is replaced with cis-4-hydroxy-L-proline and the title compound is obtained as a clear oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.70 (d, 8.3, 2H), 7.41-7.20 (m, 10H), 7.10-7.07 (m, 2H), 5.24 (dd, J = 12.1, 1H), 5.16 (dd, J = 12.2, 1H), 4.93-4.87 (m, 1H), 4.88 (bd, J = 52,4, 1H), 4.15-4.09 (m, 1H), 3.80 (ddd, J = 1.6, J = 20.7, J = 12.5, 1H), 3.37 (ddd, J = 3.1, J = 13.9, J = 36.8, 1H), 3.28 (dd, J = 13.9, J = 5.8, 1H), 3.06 (dd, J = 13.9, J = 7.2, 1H), 2.43 (s, 3H), 2.32-2.18 (m, 1H), 2.11-1.91 (m, 1H).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 170.7, 170.0, 144.4, 135.8, 135.1, 129.7, 129.4, 128.58, 128.56, 128.4, 128.1, 127.0, 91.2 (d, J = 181.0 Hz), 67.3, 60.8, 55.6 (d, J = 23.0 Hz), 53.2, 37.9, 37.0 (d, J = 22.0 Hz), 21.6.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 525 (MH &lt; + &gt;).

Example 50

Synthesis of N- (toluene-4-sulfonyl) -L- (cis-4-fluoro) -prolyl-L-phenylalanine benzyl ester

Trans-4-hydroxy-L-proline is treated with EtOH and HCl gas and the mixture is distilled to give trans-4-hydroxy-L-proline ethyl ester hydrochloride. This product is treated with TsCl and Et ₃ N in CH ₂ Cl ₂ and after aqueous workup to afford N- (toluene-4-sulfonyl) -trans-4-hydroxy-L-proline ethyl ester. This product was treated with morpholinosulfur trifluoride in CH ₂ Cl ₂ , aqueous workup and flash chromatography to N- (toluene-4-sulfonyl) -cis-4-fluoro-L-proline ethyl ester To obtain. This product was treated with NaOH in CH ₃ OH and H ₂ O, oxidized, extracted, dried over MgSO ₄ , filtered and evaporated to N- (toluene-4-sulfonyl) -cis-4-fluoro-L -Proline is obtained. This product is treated with HCl.Phe-OBn, BOP and NMM in DMF, after aqueous workup and flash chromatography, to afford the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.69 (d, 8.3, 2H), 7.41 (d, J = 7.8, 1H), 7.38-7.27 (m, 7H), 7.21-7.17 (m, 3H), 7.07-7.04 (m, 2H), 5.19 (d, J = 12.1, 1H), 5.12 (d, J = 12.1, 1H), 5.05 (dt, Jd = 52.7, Jt = 3.4, 1H), 4.89 (dt, Jd = 7.9, Jt = 6.0, 1H), 4.27 (d, J = 9.9, 1H), 3.68 (ddd, J = 1.6, J = 12.6, J = 21.1, 1H), 3.37 (ddd, J = 3.7, J = 12.4, J = 35.7, 1H), 3.13 (dd, J = 5.9, J = 13.8, 1H), 3.08 (dd, J = 6.1, J = 13.8, 1H), 2.62 (t, J = 15.9, 1H) , 2.43 (s, 3 H), 1.82-1.59 (m, 1 H).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 170.5, 169.9, 144.8, 135.5, 135.1, 132.4, 130.1, 129.41, 129.39, 128.6, 128.5, 128.4, 127.9, 126.9, 91.7 (d, J = 179.4 Hz), 67.1, 55.6 (d, J = 23.8 Hz), 36.3 (d, J = 21.5 Hz), 21.6.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 525 (MH &lt; + &gt;).

Example 51

Synthesis of N- (toluene-4-sulfonyl) -L-thiaprolyl-L-phenylalanine benzyl ester

By the procedure outlined for the preparation method of Example 3 (11), the title compound is prepared.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 2.45 (s, 3H), 3.05-3.35 (m, 2H), 4.03 (d, 1H, J = 10.3 Hz), 4.55 (d, 1H, J = 10.3 Hz ), 4.60 (m, 1H), 4.86 (m, 1H), 5.20 (dd, 2H, J = 12.1, 13.6 Hz), 7.03 (m, 1H), 7.14-7.40 (m, 12H), 7.70 (d, 2H, J = 8.2 Hz).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 22.3, 33.5, 38.1, 52.0, 54.2, 65.8, 68.0, 127.8, 128.5, 129.1, 129.2, 129.9, 130.7, 134.0, 135.6, 136.0, 145.6, 168.7, 171.1.

Mass spectroscopy: (FAB +) 525 (M + H).

Example 52

Synthesis of N- (toluene-4-sulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester

By the procedure described for the preparation of Example 3 (11), the title compound is prepared.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 1.10 (s, 6H), 2.44 (s, 3H), 3.08-3.14 (m, 2H), 3.85 (s, 1H), 4.39 (d, 1H, J = 9.7 Hz), 4.52 (d, 1H, J = 9.7 Hz), 4.94 (m, 1H), 5.17 (m, 2H), 7.01 (d, 1H, J = 2.5 Hz), 7.10-7.40 (m, 12H) , 7.74 (d, 2H, J = 8.3 Hz).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 22.3, 24.5, 29.8, 38.9, 51.2, 53.9, 55.2, 67.9, 74.1, 127.6, 128.7, 129.0, 129.1, 129.2, 129.3, 130.0, 130.5, 133.3, 135.7, 136.4, 145.3, 169.0, 171.3.

Mass spectroscopy: (FAB +) 553 (M + H).

Example 53

Synthesis of N- (2-methoxycarbonylbenzenesulfonyl) -L-prolyl-L-phenylalanine benzyl ester

Triethylamine (4.70 mmol) is added to L-prolyl-L-phenylalanine benzyl ester (4.64 mmol) in THF (15 mL) and the reaction is processed at room temperature for 30 minutes. The reaction mixture is cooled to 0 ° C., methyl 2- (chlorosulfonyl) benzoate (4.64 mmol) is added and the reaction is then processed at room temperature for 4 hours. The reaction was extracted with EtOAc (3 × 50 mL) and water (50 mL) and the combined organic layers were washed successively with saturated NaHCO ₃ (50 mL) and saturated NaCl (2 × 50 mL), dried over MgSO ₄ , filtered, Rotary distillation gives a colorless oil (2.30 g, 90%). Crude product is purified by silica gel chromatography (50% EtOAc / hexanes, R _f = 0.47) to give a colorless oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.88 (d, 1H, J = 7.29 Hz), 7.48 & 7.16 (m, 14H), 5.04 (m, 2H), 4.75 (m, 1H), 4.37 (d , 1H, 6.92 Hz), 3.86 (s, 3H), 3.40 & 3.28 (m, 2H), 3.06 (m, 2H), 1.75, 1.61 & 1.32 (m, 4H).

¹³ C NMR (CDCl ₃ , 300 MHz); δ = 171.61, 171.32, 171.23, 169.03, 136.75, 135.89, 135.51, 133.89, 133.62, 131.14, 130.34, 129.77, 129.38, 129.09, 129.00, 128.96, 128.88, 128.79, 127.48, 62.13, 60.13, 60.13, 60.13, 60.13, 60.13, 60.13. 31.07, 24.48, 21.56.

Mass spectroscopy: (+ FAB) 551 (M + H).

Example 54

Synthesis of N- (3,5-dichlorobenzenesulfonyl) -L-prolyl-L-phenylalanine

L-proline methyl ester hydrochloride (2.68 g, 16.2 mmol) is dissolved in pyridine (20 mL), 3,5-dichlorophenylsulfonyl chloride (3.57 g, 14.6 mmol) is added, and then the mixture is stirred for 19 h. do. Water (5 mL) is added and the mixture is stirred for 45 minutes before dilution with water (200 mL). The mixture is extracted with Et ₂ O (2 × 150 mL) and the combined extracts are washed with water (3 × 100 mL), 1N HCl (150 mL) and saturated aqueous NaHCO ₃ (150 mL), then dried (MgSO ₄ ) and vacuum Under filtration and evaporation to give 3,5-dichlorophenylsulfonyl-L-proline methyl ester (4.34 g, 88%) as an oil.

Methyl ester is dissolved in MeOH (20 mL) and 1N NaOH (20 mL) is added. The mixture is warmed gently until homogeneous and stirred for 1.5 hours. The solvent is removed in vacuo and the residue is dissolved in water (50 mL). The aqueous solution is washed with Et ₂ O (30 mL) and acidified with 12N HCl. The mixture was washed with CHCl ₃ (2 × 40 mL), the combined extracts were dried (MgSO ₄ ), then filtered and distilled under vacuum to give N- (3,5-dichlorophenylsulfonyl) -L-proline (3.37 g , 83%) is obtained as a solid.

3,5-dichlorophenyl-sulfonyl-L-proline is coupled with L-phenylalanine ethyl ester using the procedure described in Method 3 to afford the title compound ethyl ester (348 mg, 45%).

The title compound is prepared by hydrolysis of the ethyl ester using NaOH in MeOH. NMR analysis indicates the presence of diastereomers.

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.11 (br s, 1H), 7.72 (d, 2H, J = 1.9 Hz), 7.61 (q, 1H, J = 2.0 Hz), 7.39-7.20 (6H), 4.92 (m, 1H), 4.15 (m, 1H), 3.52-3.09 (4H), 2.11 (m, 1H), 1.95-1.49 (3H).

¹³ C NMR (CDCl ₃ ): δ = 175.4, 175.2, 171.8, 171.7, 139.5, 139.3, 137.0, 136.6, 135.8, 134.0, 134.0, 129.9, 129.5, 127.7, 126.7, 111.4, 63.1, 62.9, 53.8, 53.6, 50.5, 50.3, 45.1, 37.9, 31.3, 30.8, 24.8, 24.6.

Mass spectroscopy: FAB m / e 471 (M + H).

Example 55

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-hydroxysuccinimide ester

Equimolar amounts of Cbz-L-phenylalanine, N-hydroxysuccinimide and DCC in CH ₂ Cl ₂ are stirred at room temperature for 3 hours. Hexane is added and the reaction mixture is filtered through a celite bed. The filtrate is extracted with 10% citric acid, water and saturated NaCl, dried over MgSO ₄ , filtered and concentrated to give the title compound as a solid having a melting point of 186-188 ° C.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.66 (m, 3H), 7.28 (m, 6H), 4.64 (m, 1H), 4.01 (m, 1H), 3.36 (m, 1H), 2.89 (m , 6H), 2.42 (s, 3H), 1.90 & 1.68 (m, 3H), 1.33 & 1.15 (m, 4H).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 173.07, 171.83, 166.89, 144.86, 137.90, 133.35, 130.51, 130.44, 129.06, 128.33, 127.36, 63.21, 50.12, 47.74, 45.09, 38.30, 36.53, 34.30, 30.67, 34.30 24.454, 24.35, 22.15.

Mass spectroscopy: (+ FAB) 528 (M + H).

Example 56

Synthesis of N- (thiophene-2-sulfonyl) -L-prolyl-L-phenylalanine methyl ester

The title compound is prepared via method 14 and separated as an oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.63 (m, 1H), 7.32-7.10 (br m, 7H), 4.85 (m, 1H), 4.09 (m, 1H), 3.77 (s, 3H), 3.40 (m, 1H), 3.38 (m, 1H), 3.17 (m, 1H), 3.05 (m, 1H), 2.05 (m, 1H), 1.62-1.40 (br m, 3H).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 171.9, 170.9, 136.6, 136.2, 133.9, 133.4, 129.8, 129.1, 128.3, 127.7, 63.2, 53.9, 53.1, 38.4, 30.1, 24.8.

Mass spectroscopy: (PI-FAB) 423 (M + H) ⁺ .

Example 57

Synthesis of N- (thiophene-2-sulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared from Example 56 (285) via Method 6, and the product is separated as a solid.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.14 (d, 1H, J = 8.0 Hz), 8.02 (d, 1H, J = 5.0 Hz), 7.70 (d, 1H, J = 4.0 Hz), 7.25 (m, 6H), 4.51 (m, 1H), 4.05 (m, 1H), 3.40 (m, 1H), 3.40 (m, 1H), 3.20-2.90 (br m, 3H), 1.65-1.32 (br m, 4H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.9, 170.9, 137.8, 136.4, 134.1, 133.4, 129.7, 128.6, 128.5, 126.8, 62.1, 53.5, 49.7, 36.8, 30.7, 24.1

Mass spectroscopy: (PI-FAB) 409 (M + H) ⁺ .

Example 58

Synthesis of N- (1,3-dimethyl-5-chloropyrazole-4-sulfonyl) -L-prolyl-L-phenylalanine

Methods 14 and 7 give the title compound, and separate the product as a white solid.

NMR data are as follows:

¹ H NMR (CD ₃ OD, 300 MHz): δ = 7.00-6.93 (br m, 5H), 4.21 (m, 1H), 3.96 (m, 1H), 3.59 (s, 3H), 3.16-3.03 (br m , 3H), 2.88 (m, 1H), 2.14 (s, 3H), 1.68 (m, 2H), 1.56-1.29 (br m, 2H).

¹³ C NMR (CD ₃ OD, 75 MHz): δ = 177.9, 173.6, 150.9, 139.7, 131.8, 131.4, 129.9, 127.9, 115.3, 63.7, 57.6, 50.7, 39.4, 37.7, 32.6, 25.8, 14.9.

Mass spectroscopy: (PI-FAB) 477 (M + H) ⁺ .

Example 59

Synthesis of N- (1-phenylethanesulfonyl) -L-prolyl-L-phenylalanine

In a rapidly stirred mixture of CH ₃ Cl ₃ and H ₂ O, PhCH ₂ CH ₂ SH is treated with Cl ₂ , the CHCl ₃ layer is washed with 5% NaHSO ₃ and saturated NaCl, filtered and concentrated to give PhCH ₂ CH ₂ SO ₂ Cl is obtained. The product is treated with Pro-OtBu and Et ₃ N in CH ₂ Cl ₂ and after aqueous workup, PhCH ₂ CH ₂ SO ₂ -Pro-OtBu is obtained. Treat the product with HCO ₂ H and distill the mixture to afford PhCH ₂ CH ₂ SO ₂ -Pro-OH. The product is treated with HCl.Phe-OtBu, EDAC, HOBT and Et ₃ N in CH ₂ Cl ₂ , aqueous work-up and flash chromatography to obtain PhCH ₂ CH ₂ SO ₂ -Pro-Phe-OtBu. Treat the product with HCO ₂ H and distill the mixture to afford the title compound as a clear oil.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 8.06 (d, J = 8.1, 1H), 7.32-7.14 (m, 10H), 4.52-4.24 (m, 1H), 4.26 (dd, J = 7.0 , J = 2.6, 1H), 3.40-3.31 (m, 3H), 3.29-3.23 (m, 2H), 3.08 (dd, J = 13.8, J = 4.8, 1H), 2.97-2.90 (m, 3H), 2.05-1.99 (m, 1 H), 1.77-1.65 (m, 3 H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.7, 171.4, 138.5, 137.4, 129.2, 128.6, 128.5, 128.1, 126.5, 126.4, 60.8, 53.1, 50.0, 48.6, 36.6, 30.9, 28.6, 24.2.

Mass spectroscopy: (+ FAB, 3-nitrobenzyl alcohol) 431 (MH &lt; + &gt;).

Example 60

Synthesis of N- (1-methylimidazole-4-sulfonyl) -L-prolyl-L-phenylalanine methyl ester

Method 14 is used to prepare the title compound, which is isolated as an oil.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 7.55-7.40 (br m, 3H), 7.31-7.10 (br m, 5H), 4.80 (m, 1H), 4.31 (m, 1H), 3.70 (s, 3H), 3.49-3.18 (br m, 3H), 3.05 (m, 1H), 2.05 (m, 1H), 1.74 (m, 2H), 1.41 (m, 1H).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 172.0, 171.6, 140.1, 137.9, 136.8, 129.8, 129.0, 127.5, 126.1, 63.1, 53.9, 52.9, 50.3, 38.4, 34.6, 30.4, 24.9.

Mass spectroscopy: (PI-FAB) 421 (M + H) ⁺ .

Example 61

Synthesis of N- (1-methylimidazole-4-sulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared from Example 60 (297) using Method 7 and separated as a white solid.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 7.83 (d, 2H, J = 9.0 Hz), 7.65 (d, 1H, J = 5.0 Hz), 7.20-7.05 (br m, 5H), 4.14 ( m, 1H), 3.94 (m, 1H), 3.67 (s, 3H), 3.04 (m, 4H), 1.81 (m, 1H), 1.50 (m, 2H), 1.29 (m, 1H).

¹³ C NMR (CDCl ₃ , 75 MHz): δ = 173.1, 169.9, 140.7, 139.2, 136.0, 130.0, 127.8, 126.7, 125.9, 63.0, 55.1, 49.7, 36.8, 33.9, 30.5, 23.9.

Mass spectroscopy: (PI-FAB) 429 (M + H) ⁺ .

Example 62

Synthesis of N- (4-amidinobenzenesulfonyl) -L-prolyl-L-phenylalanine methyl ester

The product from Example 64 (313) was refluxed for 3.5 hours using ammonium acetate (0.32 mmol) in methanol (3 mL) to prepare the title compound, concentrated in vacuo to give an oil (114 mg, 96%). . Crude product is purified by silica gel chromatography (100% EtOAc, R _f = 0.01) to give a colorless oil (40 mg, 34%).

NMR data are as follows:

¹ H NMR (CD ₃ OD, 300 MHz): δ = 7.77 (m, 4H), 7.07 (m, 5H), 4.48 (m, 1H), 4.01 (m, 1H), 3.51 (s, 3H), 3.25 ( m, 1H), 3.04 (m, 3H), 2.85 (m, 1H), 1.50 (m, 4H).

¹³ C NMR (CD ₃ OD, 300 MHz): δ = 174.45, 173.58, 168.09, 143.77, 138.72, 134.54, 131.01, 130.84, 130.16, 130.11, 128.57, 63.71, 55.72, 53.45, 51.09, 38.72, 32.61, 25.97.

Mass spectroscopy: (+ FAB) 459 (M + H).

Example 63

Synthesis of N- (4-amidinobenzenesulfonyl) -L-prolyl-L-phenylalanine

The title compound is prepared from the product of Example 62 (303) using the procedure described in Method 7.

NMR data are as follows:

¹ H NMR (CD ₃ OD, 300 MHz): δ = 7.72 (m, 4H), 7.00 (m, 5H), 4.51 (m, 0.5H), 4.20 (t, 1H, J = 5.92), 3.93 (m, 1H), 3.23 (m, 0.5H), 3.14 (s, 1H), 3.04 (m, 1H), 2.85 (m, 1H), 1.48 (m, 4H).

¹³ C NMR (CD ₃ OD, 300 MHz): δ = 178.07, 173.31, 141.62, 141.36, 139.70, 138.57, 131.36, 131.05, 131.00, 130.21, 130.17, 130.13, 130.02, 129.92, 129.78, 129.71, 128.60, 128.03, 128.03 , 57.64, 51.20, 39.52, 32.36, 25.88.

Mass spectroscopy: (+ FAB) 467 (M + H), 489 (M + Na).

Example 64

Synthesis of N- (4-thiomethoxyimidatylbenzenesulfonyl) -L-prolyl-L-phenylalanine methyl ester

The product from Example 65 (314) was refluxed with acetone (3 mL) and methyl iodide (0.2 mL) for 30 minutes to prepare the title compound, and the reaction was concentrated under vacuum to give an oil (130 mg, 100%). Less than). Crude product was purified by silica gel chromatography (80% EtOAc / hexanes, R _f = 0.49) to give a yellow oil (110 mg, 99%).

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 9.37 (bs, 2H), 8.11 (d, 2H, J = 8.43 Hz), 7.96 (d, 2H, J = 8.49 Hz), 7.20 (m, 6H), 4.82 (m, 1H), 4.11 (m, 1H), 3.74 (s, 3H), 3.41 (m, 1H), 3.23 (m, 1H), 3.09 (m, 2H), 2.94 (s, 3H), 1.83 (m, 1 H), 1.62 (m, 3 H).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 186.47, 172.23, 171.65, 141.68, 136.53, 136.04, 130.32, 129.84, 129.16, 129.12, 127.74, 62.98, 53.98, 53.33, 50.48, 38.21, 31.58, 31.15, 31.15. 18.30.

Mass spectroscopy: (+ FAB) 490 (M + H).

Example 65

Synthesis of N- [4- (N-methyl) thioamidobenzenesulfonyl] -L-prolyl-L-phenylalanine methyl ester

Hydrogen sulfide is bubbled through a solution of methyl ester of Example 29 (71) (0.3 mmol) in pyridine (3 mL) and triethylamine (0.3 mL) at room temperature for 5 minutes to prepare the title compound. The flask is sealed and the reaction stirred for 18 hours at room temperature. The reaction mixture is concentrated under a stream of nitrogen. Dilute the residue with EtOAc (100 mL), successively dilute with 2N KHSO ₄ (2 × 50 mL) and saturated NaCl (50 mL), dry with MgSO ₄ , filter, rotary distillate, and yellow oil (146 mg, 100%). ). Silica gel TLC in 80% EtOAc / hexanes shows R _f of 0.06 in a single spot.

NMR data are as follows:

¹ H NMR (CDCl ₃ , 300 MHz): δ = 8.13 (d, 2H J = 11.54 Hz), 7.94 (d, 2H, J = 8.37 Hz), 7.69 (d, 2H, J = 8.43 Hz), 7.20 (m , 6H), 4.83 (m, 1H), 3.99 (d, 1H, J = 6.17 Hz), 3.77 (s, 3H), 3.34 (t, 1H, J = 6.59 Hz), 3.25 (m, 1H), 3.03 (m, 1 H), 1.87 (m, 1 H), 1.45 (m, 5 H).

¹³ C NMR (CDCl ₃ , 300 MHz): δ = 201.09, 172.29, 171.28, 144.50, 138.26, 136.37, 129.76, 129.21, 128.71, 128.21, 127.82, 62.92, 53.74, 53.31, 50.20, 38.37, 30.64, 30.68 24.68 .

Mass spectroscopy: (+ FAB) 476 (M + H).

Example 66

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-D, L-β- (1,2,4-triazol-3-yl) alanine

Equimolar β- (1,2,4-triazol-3-yl) -D, L-alanine methyl ester, containing 3 equivalents of triethyl amine in acetonitrile (0.45M), N- (toluene-4- Sulfonyl) -L-proline hemibenzeneate and benzotriazol-1-yloxytris (dimethylamino) phosphonium hesafluorophosphate are stirred under nitrogen for 16 hours at room temperature. The solvent is stripped to give a dark red oil. The oil is dissolved in ethyl acetate and washed with plenty of water, brine and saturated sodium bicarbonate solution and then once again with water. The organic phase is dried (Na ₂ SO ₄ ) and the solvent stripped to give a brown oil. The oil is hydrolyzed by the procedure described in Method 7 to give the title compound as a beige solid having a melting point of 153 to 163 ° C.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 400 MHz): δ = 8.29 (s, 1H), 7.77 (s, 1H), 7.57-7.52 (m, 2H), 7.37 (d, 1H, J = 8.1 Hz), 7.32 -7.29 (m, 2H), 7.22-7.19 (m, 1H), 4.40 (dd, 1H, J = 9.9 Hz, 4.2 Hz), 4.29-4.25 (m, 1H), 3.95-3.87 (m, 1H), 3.82-3.77 (m, 1H), 3.15-3.06 (m, 1H), 2.96-2.85 (m, 1H), 2.61-2.56 (m, 1H), 2.27 (s, 1H), 2.22 (s, 1H), 1.81 (t, 1H, J = 7.7 Hz), 1.75 (s, 3H), 1.75-1.50 (m, 1H), 1.17-1.15 (m, 1H), 1.05-0.94 (m, 1H), 0.92-0.89 ( m, 1 H).

IR (KBr, cm ⁻¹ ): 3400, 2900, 2850, 2500, 1775, 1600, 1450, 1400, 1200, 1175, 1050, 1025, 875, 830, 775, 700, 600, 575, 550.

Mass spectroscopy: (+ FAB) 428.3 (M + Na), 406.3 (M-H), 319.4, 269.4, 227.4, 205.4, 173.3, 113.12.

Example 67

Synthesis of N- (toluene-4-sulfonyl) -L-propyl-D, L-β- (2-thiazolyl) alanine

The procedure described in Method 3 is used to prepare the methyl ester of the title compound. Hydrolysis of the esters in the procedure described in Method 6 yields the title compound as a foam.

NMR dates are as follows:

¹ H NMR (DMSO-d ₆ , 400 MHz): δ = 8.36 (d, 0.5H, J = 8 Hz), 8.24 (d, 0.5H, J = 8 Hz), 7.7 (m, 3H), 7.58 (d, 1H , J = 3 Hz), 7.4 (m, 2H), 4.65 (m, 1H), 4.14 (m, 0.5H), 4.1 (m, 0.5H), 3.45 (m, 8H), 3.1 (m, 1H), 2.39 (s, 3 H), 1.75-1.4 (brd m, 4H).

IR (KBr, cm ⁻¹ ): 3425, 2900, 1730, 1660, 1625, 1525, 1510, 1440, 1340, 1160, 1080, 660, 580, 550.

Mass spectroscopy: (+ FAB) 424 ([M + H] ⁺ ), 323, 279, 237, 215, 197, 181, 149, 131, 109.

HPLC [Primesphere C-18; 40/60 Methanol / 0.01 M KH ₂ PO ₄ buffer pH = 3.5; Flow rate = 1 mL / min]: retention time for Diastereomer A (49%) = 16 minutes, retention time for Diastereomer B (51%) = 19.1 minutes.

Example 68

Synthesis of N- [4- (3-dimethylaminopropyloxy) -benzenesulfonyl] -L-propyl-L-phenylalanine

4- (methoxy) benzenesulfonyl chloride is dissolved in CH ₂ Cl ₂ and cooled to 0 ° C. in an ice bath. To the solution is added Pro-Phe-OMe (1 equiv) hydrochloride and TEA (2.2 equiv). The reaction is warmed to room temperature and stirred overnight under a stream of N ₂ . The reaction mixture is then concentrated, the residue is dissolved in EtOAc and H ₂ O, the organic phase is washed with saturated NaHCO ₃ and brine, then dried (MgSO ₄ ), filtered and concentrated to a sticky solid. To be used without further purification. The solid product is treated with BBr ₃ (3.0 equiv in CH ₂ Cl ₂ 1M, held at −78 ° C. for 1 hour and then warmed to room temperature). The reaction is then cooled in an ice bath and quenched with water. After concentration, the residue is dissolved in EtOAC and water. The organic phase is washed with brine, dried (MgSO ₄ ), filtered and concentrated to foam. The product is esterified with MeOH and HCl gas (0 ° C. to room temperature, 16 hours) and purified by column chromatography. The product is then added to a cooled THF solution of triphenylphosphine (1.10 equiv.), 3-dimethylimino-1-propanol (1.0 equiv.) And diethyl azobicarboxylate (1.10 equiv.). . The reaction temperature is maintained at 0 ° C. for 30 minutes, warmed to room temperature and then stirred overnight. The reaction mixture is concentrated, dissolved in EtOAc and washed with saturated NaHCO ₃ . The product is extracted with 0.2N citric acid and the aqueous phase is washed with EtOAc. The aqueous phase is made basic with solid NaHCO ₃ and the product is extracted with EtOAc. The organic phase is washed with brine, dried (MgSO ₄ ), filtered and concentrated. The product is purified by preparative TLC. Hydrolysis is carried out using method 7 to afford the title compound as a mixture of diastereomers and to separate as a white hygroscopic solid.

NMR data are as follows:

¹ H NMR (DMSO-d ₆ , 300 MHz): δ = 7.76 (d, 2H, J = 9.0 Hz), 7.74 (m, 1H), 7.15-7.10 (br m, 6H), 4.07 (t, 2H, J = 5.0 Hz), 3.90 (m, 2H), 3.21-2.85 (m, 4H), 2.37 (m, 2H), 2.37 (t, 2H, J = 7.0 Hz), 2.13 (s, 6H), 1.85 (m , 2H), 1.69 (m, 1H), 1.50-1.25 (m, 3H).

¹³ C NMR (DMSO-d ₆ , 75 MHz): δ = 172.3, 169.7, 162.6, 139.3, 130.3, 130.1, 130.1, 128.1, 127.9, 127.8, 125.9, 125.8, 115.3, 115.2, 66.7, 62.4, 55.8, 55.2, 49.3, 45.5, 36.96, 30.6, 27.0, 23.9.

Mass spectroscopy: (PI-FAB) 548 (M-H + Na) ⁺ .

Example 69

Synthesis of N- (4-thiocarbamoylbenzenesulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester

The title compound is prepared by the procedure of Example 65 (314).

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 8.48 (bs, 1H), 8.24 (bs, 1H), 8.00-7.97 (d, 2H), 7.78-7.75 (d, 2H), 7.29-7.09 (m, 10H) , 5.12 (m, 2H), 4.48 (m, 1H), 4.49-4.46 (d, 1H), 4.33-4.30 (d, 1H), 3.09 (m, 2H), 1.03-0.99 (d, 6H), 3.91 (s, 1 H).

¹ H NMR (CDCl ₃ ): δ = 200.5, 171.6, 169.2, 144.3, 138.4, 136.2, 135.6, 130.0, 129.2, 128.8, 128.4, 127.8, 74.1, 68.1, 55.1, 54.1, 51.1, 38.5, 29.8, 24.2, 21.7.

Example 70

Synthesis of N- (4-cyanobenzenesulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester

The title compound is prepared by the process of the schematic example 29 (71).

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 7.95-7.92 (d, 2H), 7.77-7.74 (d, 2H), 7.34-7.33 (m, 10H), 7.19-7.18 (m, 10H), 7.10-7.09 ( m, 10H), 6.97-6.95 (d, 1H), 5.15 (m, 2H), 4.91 (m, 1H), 4.47 (s, 2H), 3.10 (m, 2H), 1.19-1.11 (2s, 6H) .

¹³ C NMR (CDCl ₃ ): δ = 171.3, 168.4, 141.0, 136.3, 135.6, 133.7, 130.0, 129.2, 127.7, 117.8, 74.1, 68.0, 55.3, 53.86, 51.1, 38.7, 29.9, 24.4.

Example 71

Synthesis of N- (toluene-4-sulfonyl) -L- (thiamorpholinyl-3-carbonyl) -L-phenylalanine

Larson and Carlson's method [Actu Chemica Scan. 48: 517-525 (1994) to prepare L-thiamorpholine-5-carboxylic acid. The N- (toluene-4-sulfonyl) -L-thiamorpholine-5-carboxylic acid is prepared using the procedure described in Method 1. The title compound is prepared following the procedure described in Example 1 (9).

NMR data are as follows:

¹ H NMR (CD ₃ OD): δ = 7.49-7.04 (m, 9H), 4.62 (m, 1H), 4.47 (m, 1H), 3.88 (m, 1H), 3.12-2.36 (m, 5H), 2.18 (s, 3 H), 2.18-1.92 (m, 5 H).

¹³ C NMR (CD ₃ OD): δ = 175.1, 174.8, 170.8, 170.7, 146.2, 146.1, 138.9, 138.5, 131.8, 131.1, 130.3, 129.1, 128.7, 57.0, 56.9, 55.7, 45.5, 45.2, 38.9, 38.5 , 28.6, 28.1, 26.8, 26.9, 22.3.

Example 72

Synthesis of N- (toluene-4-sulfonyl) -L-[(1,1-dioxo) thiamorpholinyl-3-carbonyl] -L-phenylalanine

Larsson and Carlson (Acta Chemica Scan. 48: 522 (1994)] to prepare the title compound from the product of Example 71 (500).

NMR data are as follows:

¹ H NMR (CD ₃ OD): δ = 7.54 (m, 2H), 7.17-6.90 (m, 10H), 5.07 (m, 1H), 4.45 (m, 1H), 4.08 (m, 1H), 3.8- 3.3 (m, 2H), 3.09-2.6 (m, 5H), 2.22 (m, 3H), 2.11 (s, 3H).

¹³ C NMR (CD ₃ OD): δ = 175.1, 174.5, 168.5, 146.9, 139.5, 138.4, 137.8, 132.0, 131.1, 131.6, 130.2, 129.2, 128.8, 126.9, 62.6, 57.8, 57.6, 55.7, 55.5, 51.6 , 51.4, 43.8, 43.6, 43.6, 38.7, 38.5, 22.2, 22.1.

Example 73

Synthesis of N- (toluene-4-sulfonyl) -L-[(1,1-dioxo) thiamorpholinyl-3-carbonyl] -L-phenylalanine ethyl ester

The title compound is prepared by the process of the schematic example 72 (501).

NMR data are as follows:

¹ H NMR (CDCl ₃ ): δ = 7.72 (d, 1H), 7.63 (d, 1H), 7.35 (m, 7H), 7.13 (m, 2H), 6.80 (d, 0.5H), 6.69 (d, 0.5H), 5.13 (m, 0.5H), 5.00 (m, 0.5H), 4.89 (m, 0.5H), 4.77 (m, 0.5H), 4.21 (m, 2H), 3.99 (m, 1H), 3.19 (m, 2H), 3.02 (m, 2H), 2.88 (m, 2H), 2.44 (s, 3H), 1.25 (m, 3H).

¹³ C NMR (CDCl ₃ ): δ = 171.5, 171.4, 165.4, 165.1, 146.1, 136.1, 135.5, 131.2, 130.0, 130.0, 129.3, 129.2, 128.0, 127.8, 62.5, 62.4, 56.8, 56.6, 54.2, 53.9, 50.2, 49.9, 49.4, 49.3, 42.6, 42.3, 38.1, 37.8, 22.3, 14.7.

Example 74

Synthesis of N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine 2- (1-methyl-1,4-dihydropyridinyl-3-amido) ethyl ester

(S, S) -1-Methyl-3- [2- (3-phenyl-2-{[(1-toluene-4-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionyl A solution of oxy) -ethylcarbamoyl] -pyridinium iodine (3.8 g) is prepared in nitrogen degassed water (1000 mL) and acetonitrile (55 mL). At ambient temperature, a mixture of Na ₂ S ₂ O ₄ (2.82 g) and NaHCO ₃ (2.268 g) is added all at once and the reaction mixture is further stirred at room temperature for 3 hours until exposed to slow nitrogen emissions. The aqueous phase is then extracted with chloroform (3 x 200 mL), the organic extracts are combined, dried over MgSO ₄ , filtered and distilled. The residue is flash chromatographed on 200 g silica gel. Elution with 2% methanol / chloroform gave 800 mg of the title compound as a solid having a melting point of 68 to 72 ° C.

Other compounds prepared by the methods described above are shown in Table II below:

Example 95

In vitro assays to determine the binding of candidate compounds to VLA-4

In vitro assays are used to assess assays of candidate compounds for the α ₄ β ₁ integrin. Compounds that bind in this assay are used to assess VCAM-1 levels in biological samples in conventional assays (eg, competitive binding assays). This assay is sensitive at IC ₅₀ values as low as about 1 nM.

activity of α ₄ β ₁ integrin is Zur scat (Jurkat) cells [e.g., American Type Culture Collection (American Type Culture Collection) No. TIB 152, TIB 153, and CRL 8163], and to express high levels of α ₄ β ₁ integrin Measured by the interaction of human T-cell lines with soluble VCAM-1. VCAM-1 interacts with the cell surface in an α ₄ β ₁ integrin-dependent manner. Yednock, et al., J. Biol. Chem., 270: 28740 (1995).

Recombinant soluble VCAM-1 is expressed as a chimeric fusion protein comprising seven extracellular domains of VCAM-1 on the N-terminus and an IgG ₁ heavy chain constant region of human at the C-terminus. VCAM-1 fusion proteins are prepared and purified by the method of Yednock, above.

Jurkat cells are grown in RPM1 1640 supplemented with 10% fetal calf serum, penicillin, stereptomycin and glutamine as described by Yednock, above.

Jurkat cells are incubated for 30 minutes with 1.5 mM MnCl ₂ and 5 μg / mL 15/7 antibody on ice. Mn ⁺² activates the receptor and promote binding, 15/7 is α ₄ β recognizes the activated / ligand occupied type to fix the molecules in the art form of _VCAM-1 / α ₄ β 1 integrin mutual ₁ integrin It is a monoclonal antibody that stabilizes the action (Yednock et al.). Antibodies similar to the 15/7 antibodies were prepared by other investigators and can be used in this assay. Luque, et al., J. Biol. Chem. 271: 11067 (1996).

Cells are incubated for 30 minutes at room temperature with varying concentrations of candidate compounds in the range of 66-0.01 μM using standard 5-point serial dilutions. 15 μl of soluble recombinant VCAM-1 fusion protein is added to Jurkat cells and then incubated for 30 minutes on ice (Yednock et al.).

Cells were washed twice and then resuspended at 1: 200 in PE-conjugated goat F (ab ') ₂ anti-mouse IgG Fc (available from Immuntech, Westbrook, MD) and in the dark Incubate for 30 minutes on ice. Cells are washed twice and analyzed using standard fluorescence activated cell sorter ("FACS") assays as described in Yednock et al., Described above.

Compounds with an IC ₅₀ of less than about 15 μM have a binding affinity for α ₄ β ₁ .

When tested in this assay, the compounds of Examples 1-97 each have an IC ₅₀ of less than about 15 μM.

Example 96

In vitro saturation assay to determine the binding of candidate compounds to α ₄ β ₁

In vitro assays for determining plasma levels required for compound to be activated in an experimental autoimmune encephalomyelitis (“EAE”) model or other in vivo model described in the following examples are described below.

Log growth Jurkat cells are washed and resuspended in normal animal plasma containing 20 μg / mL of 15/7 antibody (described in the Examples above).

Jurkat cells were diluted twice in normal plasma samples containing known compounds at various concentrations from 66 to 0.01 μM using standard 12-point serial dilutions to standard curves, or peripheral blood from animals treated with candidate compounds. Are diluted 2-fold in plasma samples obtained from.

Cells were then incubated for 30 minutes at room temperature and washed twice with phosphate buffered saline (“PBS”) containing 1 mM calcium chloride and magnesium chloride (black medium), 2% fetal bovine plasma, respectively, to prevent binding. Remove 15/7 antibody.

The F (ab ') ₂ anti-mouse IgG Fc of phycoerythrin conjugated goats was then adsorbed for any nonspecific cross-reactivity by co-culture with 5% plasma from the animal species to be studied. Commercially available from Immunotech, Westbrook) at 1: 200 and then incubated for 30 minutes at 4 ° C. in the dark.

Cells are washed twice with assay medium and resuspended in the same medium. This was then followed by Yednock et al., J. Biol. Chem., 270: 28740 (1995)] by standard fluorescence activated cell sorter ("FACS") assay.

The data is then plotted as fluorescence for the dose, for example in a normal dose-response mode. Dosage levels generated in the flat portion of the top of the curve represent the levels required to obtain efficacy in an in vivo model.

Further, this test is different integrins: It can be used to measure the plasma level required to saturate the binding sites (for example, α ₄ β ₁ and the α ₉ β ₁ integrin closest integrins) [see:. Palmer et al, J Cell Bio. 123: 1289 (1993). Such binding may include, for example, airway hyperreactions and obstructions that occur with chronic asthma, smooth muscle cell proliferation in atherosclerosis, vascular occlusion after angioplasty, fibrosis and glomerular wounds as a result of kidney disease, aortic stenosis, rheumatoid It is believed to be useful in vivo for inflammatory conditions mediated by α ₉ β ₁ integrin, including synovial hypertrophy in arthritis, and inflammation and wounds that occur with ulcerative colitis and Crohn's disease.

Thus, to the assay measuring the binding of the α ₉ β ₁ integrin described above, Jour scat cells instead of the integrin α ₉ [see:. Yokosaki et al, J. Biol. Chem., 269: 26691 (1994)], can be performed with SW 480 (ATCC # CCL228), a human colon carcinoma cell line transfected with cDNA encoding. In contrast, SW 480 cells expressing other α and β ₁ subunits can be used.

Accordingly, another aspect of the invention relates to a method of treating a disease mediated by α ₉ β ₁ in a patient, including administering to the mammalian patient a therapeutically effective amount of a compound of the invention. Such compounds are preferably administered in the pharmaceutical compositions described herein. The effective daily dosage depends on the age, weight and condition of the patient, which factors are easily determined by the attending physician. However, in a preferred embodiment, the compound is administered at about 20-500 μg / kg per day.

Example 97

In Vivo Evaluation-Experimental Autoimmune Encephalomyelitis

Experimental autoimmune (or allergic) encephalomyelitis (“EAE”), a standard multiple sclerosis model, is used to determine the effect of candidate compounds in reducing motor muscle damage in rats or guinea pigs. Reduction of motor muscle damage is based on blocking adhesion of leukocytes and endothelial cells and correlates with anti-inflammatory activity in candidate compounds. Such models have already been described in the literature (Keszthelyi et al., Neurology, 47: 1053-1059 (1996)) and measure the delay in onset of disease.

The brains and spine of mature Hartley guinea pigs are homogenized in an equivalent volume of phosphate buffered saline. Isoform Freund's complete adjuvant (100 mg of mycobacterium tuberculosis + 10 mL Freund's incomplete adjuvant) is added to the homogenate. The 20 mL with the peripheral pump is emulsified by circulating through the syringe for about 20 minutes repeatedly.

Female Lewis rats (2-3 months of age, 170-220 g) or Hartley guinea pigs (20 days of age, 180-200 g) are anesthetized with isoflurane and 0.1 mL of emulsion injection each is injected into the flanks three times. Motor muscle injury appears to begin after approximately 9 days.

Candidate compound treatment begins on day 8, just before symptoms develop. The compound is administered subcutaneously ("SC"), orally ("PO") or intraperitoneally ("IP"). Dosage is twice daily for 5 days in the range of 10 to 200 mg / kg using conventional dosages of SC 10 to 100 mg / kg, PO 10 to 50 mg / kg and IP 10 to 100 mg / kg. Administration.

Antibody GG5 / 3 against α ₄ β ₁ integrin (Keszthelyi et al., Neurology, 47: 1053-1059 (1996)), which delays the development of symptoms, is used as a positive control for 8 and 11 days. Subcutaneously injected at day 3 mg / kg.

Weight and exercise muscle damage are measured daily. Motor muscle damage is assessed by the following clinical scores:

0 no change

1 tail weakening or paralysis

2 hind limb weakness

3 hind limb paralysis

4 death or death

If a candidate compound delays the onset of symptoms, i.e. if the clinical score does not exceed 2 or there is little weight loss compared to the control, it is considered to be active.

Example 98

In Vivo Assessment-Asthma

Inflammatory conditions mediated by α ₄ β ₁ integrins include, for example, airway hyperreactions and obstructions that occur with chronic asthma. Below, an asthma model is described that can be used to study the in vivo effects of the compounds of the present invention for use in treating asthma.

Abraham et al., J. Clin., Both of which are hereby incorporated by reference in their entirety. Invest. 93: 776-787 (1994), Abraham et al., Am J. Respir Crit Care Med. 156: 696-703 (1997), the compounds of the present invention are formulated with aerosol and administered in sheep for the Ascaris suum antigen. Compounds that reduce early antigen-induced bronchial responses and / or block late stage airway responses, such as compounds that have a protective effect against antigen-induced late response and airway hyperresponsiveness (“AHR”), are in this model. It is considered to be active.

The airway effects of candidate compounds are studied using allergic sheep (sheep) that have been shown to develop both early and late bronchial responses to inhaled Ascaris suum. After local anesthesia by passing 2% of lidocaine intranasally, a balloon catheter enters the nostrils and down the esophagus. An endotracheal tube equipped with a flexible fiber optic bronchoscope is inserted as a guide through the other nostril of the animal.

Pleural pressure is measured according to Abraham, 1994. Aerosols (see formulation below) are prepared using disposable medical dispersers that provide aerosols having a mass median aerodynamic diameter of 3.2 μm, as measured by the Anderson cascade impactor. The disperser is connected to a dosimeter consisting of a solenoid valve and an extruded air (20 psi) source. The outlet of the disperser is directed to a plastic T-piece with one end connected to the inlet of the piston respirator. The solenoid valve is activated for 1 second at the start of the inhalation cycle of the respirator. The aerosol is delivered at 20 breaths / minute at 500 mL of V _T. Only 0.5% sodium bicarbonate solution is used as a control.

To assess bronchial reactivity, an accumulated concentration-response curve for carbacol can be generated according to Abraham, 1994. Bronchial biopsies can be performed immediately before and after treatment and 24 hours after antigen challenge. Bronchial biopsies can be performed according to Abraham, 1994.

In vitro adherence studies of alveolar macrophages can also be performed according to Abraham, 1994, and calculate adherent cells (%).

Aerosol Formulations

A solution of the candidate compound at a concentration of 30.0 mg / mL in 0.5% sodium bicarbonate / brine (w / v) is prepared using the following procedure.

A. Preparation of 0.5% Sodium Bicarbonate / Brine Stock: 100.0 mL

ingredient g / mL Final concentration Sodium bicarbonate 0.5g 0.5% Brine Make enough to make 100.0 mL 100% added in sufficient quantities

fair:

1. Add 0.5 g sodium bicarbonate to a 100 mL volumetric flask.

2. Add about 90.0 mL of brine and sonicate until dissolved.

3. Add enough brine to make 100 mL and mix thoroughly.

B. Preparation of 30.0 mg / mL Candidate Compound: 10.0 mL

ingredient g / mL Final concentration Candidate compounds 0.300g 30.0 mg / mL 0.5% Sodium Bicarbonate / Brine Stock Make enough to make 10.0 mL 100% by adding sufficient amount

fair:

1. Add 0.300 g of the candidate compound to a 10.0 mL volumetric flask.

2. Add about 9.7 mL of 0.5% sodium bicarbonate / brine stock solution.

3. Sonicate until the candidate compound is completely dissolved.

4. Add enough 0.5% sodium bicarbonate / brine to 10.0 mL, and mix thoroughly.

When using conventional oral formulations, the compounds of the present invention are active in this model.

Claims (14)

Compounds of formula (I) and pharmaceutically acceptable salts thereof. Formula I In Formula I above, R ¹ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclic, substituted heterocyclic, heteroaryl and substituted heteroaryl, R ² and R ³ may form a saturated heterocyclic group or a saturated substituted heterocyclic group together with a nitrogen atom bonded to R ² and a carbon atom bonded to R ³ , provided that when mono-substituted, Substituents in such saturated heterocyclic groups are not carboxy, R ⁵ is a - is selected from the group consisting of heteroaryl (wherein, n is an integer from 1 to _{4), - (CH 2)} n - aryl and - (CH _{2) n} Q is —C (X) NR ⁷ —, where R ⁷ is selected from the group consisting of hydrogen and alkyl, X is selected from the group consisting of oxygen and sulfur, Provided that R ¹ is 2,4,6-trimethylphenyl, R ² and R ³ together with the pendant nitrogen atom and carbon atom form a pyrrolidinyl ring, and when Q is -C (O) NH-, R ⁵ Is not benzyl. Compounds of Formula (IA) and pharmaceutically acceptable salts thereof. Formula IA In Formula IA above, R ¹ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, heterocyclic, substituted heterocyclic, heteroaryl and substituted heteroaryl, R ² and R ³ may form a saturated heterocyclic group or a saturated substituted heterocyclic group together with a nitrogen atom bonded to R ² and a carbon atom bonded to R ³ , provided that when mono-substituted, Substituents in such saturated heterocyclic groups are not carboxy, R ⁵ is a - is selected from the group consisting of heteroaryl (wherein, n is an integer from 1 to _{4), - (CH 2)} n - aryl and - (CH _{2) n} R ⁶ is amino, alkoxy, substituted alkoxy, cycloalkoxy, substituted cycloalkoxy, -O- (N-succinimidyl), -NH-adamantyl, -O-cholest-5-ene-3-β -Yl, -NHOY, where Y is hydrogen, alkyl, substituted alkyl, aryl and substituted aryl, -NH (CH ₂ ) _P COOY, where p is an integer from 1 to 8 and Y is defined above ), -OCH ₂ NR ⁹ R ¹⁰ where R ⁹ is selected from the group consisting of -C (O) -aryl and -C (O) -substituted aryl, R ¹⁰ is hydrogen and -CH ₂ COOR ¹¹ (wherein R ¹¹ is alkyl) and -NHSO ₂ Z, wherein Z is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, Substituted heteroaryl, heterocyclic, and substituted heterocyclic), Q is —C (X) NR ⁷ —, where R ⁷ is selected from the group consisting of hydrogen and alkyl, X is selected from the group consisting of oxygen and sulfur, Provided that R ¹ is 4-methylphenyl, R ² and R ³ together with the pendant nitrogen and carbon atoms form a pyrrolidin-2-yl ring, R ⁵ is benzyl and Q is —C (O) When NH-, R ⁶ is not —NH (CH ₂ ) ₂ CO ₂ Et or — (1R, 2S, 5R)-(−)-methyl ester; B: R ¹ is 4-methylphenyl, R ² and R ³ together with the pendant nitrogen atom and carbon atom form a 3-β-phenyl-D-pyrrolidin-2-yl ring, R ⁵ is benzyl, When Q is -C (O) NH-, R ⁶ is not -OCH ₂ CH ₃ , C: R ¹ is 1-N-methyl-3-methyl-5-chloropyrazol-4-yl, and R ² and R ³ together with the pendant nitrogen and carbon atoms form a pyrrolidin-2-yl ring When R ⁵ is benzyl and Q is -C (O) NH-, R ⁶ is not -OCH ₃ ; D: R ¹ is 4-methylphenyl, R ² and R ³ together with the pendant nitrogen atom and carbon atom form a pyrrolidin-2-yl ring, R ⁵ is D-benzyl and Q is -C (O ) NH-, R ⁶ is not -OCH ₂ CH ₃ , E: R ¹ is 4-methylphenyl, R ² and R ³ together with the pendant nitrogen and carbon atoms form a 5,5-dimethyl-1,1-dioxo-thiaprolyl ring, R ⁵ is benzyl When Q is -C (O) NH-, R ⁶ is not -OC (CH ₃ ) ₃ . The compound of claim 1 or 2, wherein R ¹ is selected from the group consisting of aryl, substituted aryl, heterocyclic, substituted heterocyclic, heteroaryl, and substituted heteroaryl. The compound of claim 1 or 2, wherein R ¹ is 4-methylphenyl, methyl, benzyl, n-butyl, 4-chlorophenyl, 1-naphthyl, 2-naphthyl, 4-methoxyphenyl, phenyl, 2, 4,6-trimethylphenyl, 2- (methoxycarbonyl) phenyl, 2-carboxyphenyl, 3,5-dichlorophenyl, 4-trifluoromethylphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl , 4- (CH ₃ C (O) NH-) phenyl, 4-trifluoromethoxyphenyl, 4-cyanophenyl, isopropyl, 3,5-di- (trifluoromethyl) phenyl, 4-t- Butylphenyl, 4-t-butoxyphenyl, 4-nitrophenyl, 2-thienyl, 1-N-methyl-3-methyl-5-chloropyrazol-4-yl, phenethyl, 1-N-methyl Midazol-4-yl, 4-bromophenyl, 4-aminophenyl, 4-methylamidinophenyl, 4- [CH ₃ SC (= NH)] phenyl, 5-chloro-2-thienyl, 2,5 -Dichloro-4-thienyl, 1-N-methyl-4-pyrazolyl, 2-thiazolyl, 5-methyl-1,3,4-thiadiazol-2-yl, 4- [H ₂ NC (S )] Phenyl, 4-aminophenyl, 4-fluorophenyl, 2-fluorophenyl, 3-fluorophenyl, 3,5-difluorophenyl, Naphthyridin-3-yl, pyrimidin-2-yl, 4- (3'-dimethyl amino -n- propoxy) compound is selected from the group consisting of phenyl. The compound according to claim 1 or 2, wherein R ² and R ³ are selected from the group consisting of nitrogen, oxygen and sulfur together with the nitrogen atom bonded to the R ² substituent and the carbon bonded to the R ³ substituent. To heterocyclic groups or substituted heterocyclic groups of 4 to 6 ring atoms having 3 heteroatoms in the ring, the ring optionally being fluoro, methyl, hydroxy, amino, phenyl, thiophenyl and thiobenzyl From 10 to having one or two heteroatoms selected from nitrogen, oxygen, and sulfur atoms, substituted with one or two substituents selected from the group consisting of, or fused with another saturated heterocyclic, or cycloalkyl ring such as a cyclohexyl ring; A compound capable of providing a fused ring heterocycle of 14 ring atoms. ^{3. The} compound of claim 1, wherein the R ² and R ³ heterocyclic rings are azetidin-2-yl, thiazolidin-4-yl, piperidin-2-yl, piperidin-2-yl, thio A compound selected from the group consisting of morpholin-3-yl and pyrrolidin-2-yl. ^{3. The} compound of claim 1, wherein the R ² and R ³ substituted heterocyclic rings are 4-hydroxypyrrolidin-2-yl, 4-fluoropyrrolidin-2-yl, 3-phenylpyrroli Din-2-yl, 3-thiophenylpyrrolidin-2-yl, 4-aminopyrrolidin-2-yl, 3-methoxypyrrolidin-2-yl, 4,4-dimethylpyrrolidin-2 -Yl, 4-N-Cbz-piperidin-2-yl, 5,5-dimethylthiazolidin-4-yl, 1,1-dioxo-thiazolidin-4-yl, L-1,1- A compound selected from the group consisting of dioxo-5,5-dimethylthiazolidin-4-yl and 1,1-dioxothiomorpholinyl. According to claim 1 or 2, wherein, R ⁵ is benzyl, phenethyl, -CH ₂ - (3- indolyl), -CH ₂ - (1- naphthyl), -CH ₂ - (2- naphthyl) , -CH ₂ - (2- thienyl), -CH ₂ - (3- pyridyl), -CH ₂ - (5- imidazolyl), -CH ₂ -3- (1,2,4- triazolyl ), and -CH ₂ - a compound selected from the group consisting of (2-thiazolyl). The compound of claim 2, wherein R ⁶ is methoxy, ethoxy, iso-propoxy, n-butoxy, t-butoxy, cyclopentoxy, neo-pentoxy, 2-α-iso-propyl-4-β -Methylcyclohexyl, 2-β-isopropyl-4-β-methylcyclohexyl, -NH ₂ , benzyloxy, -NHCH ₂ COOH, -NHCH ₂ CH ₂ COOH, -NH-adamantyl, -NHCH ₂ CH ₂ COOCH ₂ CH ₃ , -NHSO ₂ -p-CH ₃ -ψ, -NHOR ⁸ , wherein R ⁸ is hydrogen, methyl, iso-propyl or benzyl, O- (N-succinimidyl), -O-cholest-5-en-3-β-yl, -OCH ₂ -OC (O) C (CH ₃ ) ₃ , -O (CH ₂ ) zNHC (O) W where z is 1 or 2 And W is selected from the group consisting of pyrid-3-yl, N-methylpyridyl and N-methyl-1,4-dihydro-pyrid-3-yl) and -NR "C (O) -R Wherein R 'is aryl, heteroaryl or heterocyclic, and R "is hydrogen or -CH ₂ C (O) OCH ₂ CH ₃ . N- (methanesulfonyl) -L-prolyl-L-phenylalanine, N- (α-toluenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prop Rollyl-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L- (N-methyl) phenylalanine, N- (toluene-4-sulfonyl) -L-pipecolinyl-L-phenylalanine, N- (toluene-4-sulfonyl) -D-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L- (4-hydroxy) prolyl-L-phenylalanine, N- (toluene 4-sulfonyl) -L-prolyl-D, L-homophenylalanine, N- (4-chlorobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (1-naphthalenesulfonyl) -L -Prolyl-L-phenylalanine, N- (2-naphthalenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-methoxybenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-t-butylbenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L- (4-fluoro) prolyl-L-phenylalanine, N- (n -Butanesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine, N- (2-methoxycarbonylbenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (2-carboxybenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl ) -L-thiaprolyl-L-phenylalanine, N- (3,5-dichlorobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-trifluoromethoxybenzenesulfonyl) -L- Prolyl-L-phenylalanine, N- (3,4-dichlorobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -D, L- (3-phenyl) prop Rollyl-L-phenylalanine, N- (3,4-dimethoxybenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-nitrobenzenesulfonyl) -L-prolyl-L-phenylalanine, N -(4-acetamidobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-cyanobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sul Ponyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-β- (1-naphthyl) -L-alanine, N- (toluene-4-sulfonyl ) -L-prolyl-β- (2-naphthyl) -L-alanine, N- (toluene-4-sulfonyl) -L-prolyl-β- (2-thienyl) -L-alanine, N- (isopropanesulfonyl) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-β- (3-pyridyl) -L-alanine, N- ( Toluene-4-sulfonyl) -L- (4-phenylthio) prolyl-L-phenylalanine, N- (toluene-4-sulfonyl)-(4-benzylthio) -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L-histidine, N- (toluene-4-sulfonyl) -L- (4-amino) prolyl-L-phenylalanine, N- (toluene- 4-sulfonyl) -L-prolyl-L-phenylalanineamide, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L -Prolyl-L-phenylalanine ethyl ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-methoxyamide, N- (toluene-4-sulfonyl) -L-prolyl -L-phenylalanine N-benzyloxyamide, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N- (toluene-4-sulfonyl) amide, N- (toluene-4-sulfonyl ) -L-Prolyl-L-phenylalanine-β-alanine, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine-N- Idroxyamide, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine isopropyl ester, N- (toluene-4-sulfonyl) -L- (4-hydroxy) prolyl-L -Phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanyl- (N-benzoyl) glycine ethyl ester, N- (toluene-4-sulfonyl) -L- (4- Fluoro) prolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L-thiaprolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L- (5 , 5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine ethyl ester, N- (2- Methoxycarbonylbenzenesulfonyl) -L-prolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L- (3-phenyl) prolyl-L-phenylalanine ethyl ester, N- ( Toluene-4-sulfonyl) -L- (4-methoxy) prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine (1S, 2R, 5S)- (+)-Methyl ester, N- ( Toluene-4-sulfonyl) -L-prolyl-L-phenylalanine N-hydroxysuccinimide ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine 2- (nicotinamido ) Ethyl ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine 2- (1-methylpyridinium-3-amido) ethyl ester, N- (toluene-4-sulfonyl) -L-Prolyl-L-phenylalanine cholesteryl ester, N- (toluene-4-sulfonyl) -L-prolyl-L-phenylalanine 2- (1-methyl-1,4-dihydropyridinyl-3 -Amido) ethyl ester, N- (thiophen-2-sulfonyl) -L-prolyl-L-phenylalanine methyl ester, N- (thiophen-2-sulfonyl) -L-prolyl-L-phenylalanine , N- (5-chloro-1,3-dimethylpyrazole-4-sulfonyl) -L-prolyl-L-phenylalanine, N- (2-phenylethanesulfonyl) -L-prolyl-L-phenylalanine , N- (1-methylimidazole-4-sulfonyl) -L-prolyl-L-phenylalanine methyl ester, N- (1-methylimidazole-4-sulfonyl) -L-prolyl-L -Phenylalanine, N- (4-aminobenzenesulfonyl) -L-prop Aryl-L-phenylalanine methyl ester, N- (4-aminobenzenesulfonyl) -L-prolyl-L-phenylalanine, N- (4-thiomethoxyimidatylbenzene-4-sulfonyl) -L-prolyl -L-phenylalanine methyl ester, N- [4- (N-methylthioamido) benzenesulfonyl] -L-prolyl-L-phenylalanine methyl ester, N- (toluene-4-sulfonyl) -L-prop Rollyl-D, L-β- (1,2,4-triazol-3-yl) alanine, N- (toluene-4-sulfonyl) -L-proline-D, L-β- (thiazole-2 -Yl) alanine, N- [4- (3-dimethylaminopropoxy) benzenesulfonyl] -L-prolyl-L-phenylalanine, N- (toluene-4-sulfonyl) -L-pyrrolidine-2 -Yl-thiocarbonyl-L-phenylalanine, N- (4-thiocarbamoylbenzenesulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester, N- (4-cyano Benzenesulfonyl) -L- (5,5-dimethyl) thiaprolyl-L-phenylalanine benzyl ester, N- (toluene-4-sulfonyl) -L-prolyl-D-phenylalanine, N- (toluene-4 -Sulfonyl) -L- (thiamorpholine-3-carbonyl) -L-phenylalanine, N- (toluene-4- Ponyl)-[(1,1-dioxo) thiamorpholine-3-carbonyl] -L-phenylalanine, N- (toluene-4-sulfonyl) -L- (3,3-dimethyl) prolyl-L -Phenylalanine, N- (toluene-4-sulfonyl) -L- (5,5-dimethyl-1,1-dioxo) thiaprolyl-L-phenylalanine, N- (toluene-4-sulfonyl)-[ (1,1-dioxo) thiamorpholine-3-carbonyl] -L-phenylalanine ethyl ester, N- (toluene-4-sulfonyl) -L-proline-L-phenylalanine t-butyl ester, N- ( Toluene-4-sulfonyl) -L-pyrrolidin-2-yl-thiocarbonyl-L-phenylalanine methyl ester, N- (benzenesulfonyl) -L-prolyl-L-phenylalanine methyl ester, N- ( Toluene-4-sulfonyl) -L- (5-oxo) prolyl-L-phenylalanine ethyl ester, N- (toluene-4-sulfonyl) -L- (5-oxo) prolyl-L-phenylalanine; And pharmaceutically acceptable salts thereof, as well as one ester consisting of methyl ester, ethyl ester, n-propyl ester, isopropyl ester, n-butyl ester, isobutyl ester, secondary butyl ester and tertiary butyl ester A compound selected from the group consisting of any of the ester compounds mentioned above, substituted with other esters selected from the group. A method of binding VLA-4 to a biological sample, including contacting the compound with a biological sample under conditions such that the compound of claim 1 or 2 binds to VLA-4. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 or 2 and a pharmaceutically acceptable carrier. A method of treating an inflammatory disease in a patient mediated by VLA-4, comprising administering the pharmaceutical composition of claim 12 to the patient. The method of claim 13, wherein the inflammatory disease is asthma, Alzheimer's disease, atherosclerosis, AIDS dementia, diabetes (including acute pediatric diabetes), inflammatory bowel disease (including ulcerative colitis and Crohn's disease), multiple sclerosis, rheumatoid arthritis, tissue Selected from the group consisting of transplantation, tumor metastasis, meningitis, encephalitis, ataxia and other cerebral trauma, nephritis, retinitis, atopic dermatitis, psoriasis, myocardial ischemia, and acute leukocyte-mediated lung injury such as those occurring in adult respiratory distress syndrome Way.