CA2242877A1 - Integrin receptor antagonists - Google Patents
Integrin receptor antagonists Download PDFInfo
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- CA2242877A1 CA2242877A1 CA002242877A CA2242877A CA2242877A1 CA 2242877 A1 CA2242877 A1 CA 2242877A1 CA 002242877 A CA002242877 A CA 002242877A CA 2242877 A CA2242877 A CA 2242877A CA 2242877 A1 CA2242877 A1 CA 2242877A1
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- butanoyl
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/72—Nitrogen atoms
- C07D213/73—Unsubstituted amino or imino radicals
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
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- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D213/54—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/55—Acids; Esters
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D213/54—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/56—Amides
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/62—Oxygen or sulfur atoms
- C07D213/63—One oxygen atom
- C07D213/68—One oxygen atom attached in position 4
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/72—Nitrogen atoms
- C07D213/74—Amino or imino radicals substituted by hydrocarbon or substituted hydrocarbon radicals
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- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/72—Nitrogen atoms
- C07D213/75—Amino or imino radicals, acylated by carboxylic or carbonic acids, or by sulfur or nitrogen analogues thereof, e.g. carbamates
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- C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
- C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
- C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D277/38—Nitrogen atoms
- C07D277/40—Unsubstituted amino or imino radicals
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- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/04—Ortho-condensed systems
Abstract
Fibrinogen receptor antagonists having formula (I), for example (a), which are useful for inhibiting the binding of fibrinogen to blood platelets and for inhibiting the aggregation of blood platelets.
Description
W O 97~2625n PCTrUS97/00572 TITLE OF THE INVENTION
INTEGRIN l~ECEPTOR ANTAGONISTS
BACKGROUND OF THE INVEN~ION
S The invention relates generally to modulating cell adhe,c;ionand to inhibiting the binding of fibrinogen and other proteins to blood plate~ets, and inhibiting the aggregation of blood platelets specifically to the gp IIb/IIIa fibrinogen receptor ,site. Fibrinogen is a glycoprotein present in blood plasma that participates in platelet aggregation and in fibrin formation. Platelets are cell-like anucleated *agments, found in the blood of al} m~mm~ls, that al.~io participate in blood coagulation.
Inter~action o~ fibrinogen with the IIb/~Ila receptor site is known to be essential for normal platelet function.
When a blood vessel is damaged by an injury or other causative factor, platelets adhere to the disrupted subendothethial surface. The adherent platelets subse4uently release biologically active constituents and aggregate. Aggregation is initiated by the binding of agonists, such as thrombin, epinephrine, or ADP to specific platelet membrane receptors. Stimulation by agonists results in exposure of latent fibrinogen receptors on the platelet surface, and binding of fibrinogen to the glycoprotein IIb/IIIa receptor complex.
Attempts have been made to use natural product~; and synthetic peptides to determine the mechanism of adhesion and platelet aggregation. For example, Rouslahti and Pierschbacher in Sci~nce, 238, 491-497 (1987), describe adhesive proteins such as fibronectin, vitronectin, osteopontin, collagens, thrombospondin, fibrinogen, and von Willebrand factor that are present in extracellular matrices and in blood. The protein.s contain the tripeptide arginine-glycine-aspartic acid (RGD) as their glycoprotein IIb/IIIa recognition site. These arginine-glycine-aspartic acid cont~ining tripeptides are recognized by at least one member of a family of structurally related receptors, integrins, which are heterodimeric proteins with two membrane-spanning subunits. The authors state that the conformation of the tripeptide sequence in the individual proteins may be critical to recognition ~pecificity.
Cheresh in Proc. Nat'l Acad. Sci. U.S.A., 84, 6471-6475, "
(19~7), describes an Arg-Gly-Asp directed adhesion receptor expressed by human endothethial cells that is structurally similar to the rIb/IIIa complex on platelet,s but is antigenically and functionally distinct. This receptor is directly involved in endothelial cell attachment to fibrinogen, von Willebrand factor, and vitronectin.
Pier.schbacher and Rouslahti, in J. of Biol. Chem., 262, (36), 17294- 1729~i (19~7) hypothesized that the Arg-Gly-Asp sequence alone would be a sufficient signal for receptor recognition and binding and that, therefore, the conforrnation of the tri-peptide sequence would be determinative. Various synthetic peptides were produced and the authors concluded that the sterochemical con~orrnation of ~rg-Gly-Asp as influenced by enantiomeric substitutions or additions to this ,sequence signi~lcantly influenced receptor-ligand interaction. The authors furlher showed that cyclization of a decapeptide by forming a disulfide bridge between non-terminal residues Pen and Cys, rendered the peptide much less effective at inhibiting attachment to fibronectin.
In Proc. Nat'l Acad. Sci. U.S.A., 81, 59g5-59~8 (19~4), the same author.s describe tetrapeptide variants of the cell recognition site of fibronectin that retain attachment-promoting activity. Peptides having a tetrapeptide recognition site are described in U.S. Pat. Nos. 4,5~9,g81 and 4,614,517. A number of large polypeptide fragments in the cell-binding domain of fibronectin have cell-attachment activity. For example, see U.S. Pat. Nos. 4,517,686, 4,661,111 and U.S. Pat. No.
4,57~,079.
Ruggeri et al., Proc. Nat'l Acad. Sci. U.S.A., 83, 570~s-5712 (19~S6) explore a serie~s of synthetie peptides designed in lengths to 16 residues, that contain ~GD and a valine attached to the aspartic acid residue of RGD that inhibit fibrinogen binding to platelets. See also Koc~ewiak et al., Biochem., 23, 1767-1774 ~19~4); Ginsberg et al., WO 97/26250 PCTrUS97/00572 J. Bivl. Ch~m., 260(7), 3931-3936 (1985); and Haverstick et al., Bloocl, ~6(4), 946-952 (1985). Other inhibitor~ are disclosed in Eur. Pat. App.
Nos. 275,74g and 298,820.
A number of low molecular weight polypeptide factors 5 have been i,solated from ~nake venom. These ~actors apparently have high affinity for ~e gp ~Ib/~IIa complex. For example, Huang et al., J.
B~ol C~em., 262, 16157-16163 (1987); Huang et al., Bic~che~7list)y, 28, 661-1566 (1989) describe the primary structure of the venom trigramin which is a 72 amino acid polypeptide that contains the RGD subunit.
10 Echixtatin is another compound which has high affinity for the gp IIb/Il[Ia complex. This polypeptide contains 49 amino acids and has the RGI~l subunit and various disulfide bridges. Gan et al., J. Biol. Chem., 263, 19827-19g32 (1988). See also, Dennis ~t al., Proc. Nat'l Acad. Sci.
USA. 87, 2471-2475 (1989). However, the,~ie snake venom factor.~ also 15 have high affinity for other members of the adhesive protein receptor famihy including the vitronectin and fibronectin receptors so are not selective for the gp IIb/IIIa complex.
While it is known that the tripeptide sequence Arg-Gly-Asp i,s present in certain polypeptides that can duplicate or inhibi~ the cell 20 attachment-promoting effect~ of fibronectin and vitronectin, the tripeptide Arg-Gly-Asp has low activity. At present, there is Iittle understanding of how other amino acid~ coupled to this .~equence influence binding specificity. U.S. Pat. No 5,023,233, assigned to Merck & Co., Inc., discloses small cyclic hexapeptides which contain the 25 sequence Arg-Gly-Asp and are useful platelet aggregation inhibitors.
U.S. Pat. No. 5,037,808 discloses the use of indolyl platelet-aggregation inhibitorx which are believed to act by antagonizing interactions between ~ibrinogen and/or extracellular matrix proteins and the platelet gp IIb/llIa receptor. U.S. Pat. No. 5,037,~08 disclol~;es guanidino peptide 30 mimetic compounds that retain an Asp residue which inhibit platelet aggregation. The application PCT/US90/02746 describes the use of antibody-poly-peptide conjugates wherein said polypeptides contain the Arg-Gly-Asp (RGD) sequence.
W O 97/26250 PCT~US97/00572 The application PCT/US91/00564 discloses the use of large cyclic peptides containing RGD flanked by proline residues which are platelet aggregation inhibitors. The application PCT/US90/03788 discloses small cyclic platelet aggregation inhibitors which are synthetic 5 cyclic pentapeptides containing the tripeptide sequence Arg-Gly-Asp and a thioether linkage in the cycle. The application PCT/US90/05367 published May 2, 1991 also discloses the use of peptides and pseudopeptides such as N-amidino-piperidine-3-carboxylglycyl-L-aspartyl-L-valine that inhibit platelet aggregation and thrombus 10 formation in m~mmalian blood. The application Eur. Pat. App. No.
91103462.7 discloses linear compounds which can include internal piperazinyl or piperidinyl derivatives. Eur. Pat. App. No. 91300179.~, assigned to Merck & Co., Inc., and published on July 17, 1991 discloses linear polypeptide fibrinogen receptor antagonists. Eur. Pat. App. No.
1~ 90101404.3 discloses compounds of the Rl-A-(W)a-X-(CH2)b-(Y)c-B-Z-COOR wherein Rl is a guandidino or amidino moiety and A and B
are chosen from specific monosubstituted aryl or heterocyclic moieties.
While a multitude of compounds or peptide analogs believed to inhibit platelet aggregation by inhibiting binding to a blood 20 platelet by fibrinogen are known, the present invention provides novel fibrinogen receptor antagonists that have significant binding activity and are, therefore, useful for the reasons stated herein. A number of very serious diseases and disorders involve hyperthrombotic complications which lead to intravascular thrombi and emboli. Myocardial infarction, 2~ stroke, phlebitis and a number of other serious conditions create the need for novel and effective fibrinogen receptor antagonists.
SIIMMARY OF THE INVENTION
The invention is a compound of the formula:
1~l ~ R4 X~(CH2)m~Y~(CH2)n~C~N~CH2~C~NH~CH-CH-CO2R6 3() 1 3 R~
W O 97/262~0 PCT~US97/00572 and pharmaceutically acceptable salts thereof, wherein xi,s a 5- or 6-membered monocyclic aromatic ring ~ystem containing 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either S unsubstituted or substituted with R l or R2, or a 9- to 10-membered polycyclic ring ~ystem, wherein one or more of the ring~ is aromatic, cont~ning 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either un.~ubstituted or substituted with R l or R2, wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, Cl -10 alkyl, C3-8 cycloalkyl, aryl, aryl Cl ~s alkyl, amino, amino C l ~ alkyl, C1 3 acylamino, C1 3 acylamino C1 8 alkyl, C 1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, 2~ C 1-6 dialkylamino, C1 6 dialkylamino Cl ~ alkyl, C 1 4 alkoxy, CI 4 alkoxy C1 6 alkyl, carboxy, carboxy C1 6 alkyl, Cl 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy C1 6 alkyloxy and W O 97/26250 PCT~US97/00572 hydroxy, hydroxy C1 6 alkyl;
Y i.s O N H R6 Rs -(CH2)0 6- . -C-C-, -C=C~, - C -, - C - ~ -N - , -0-, - S02NH - , - NHS02 -, -N-~- -C ~N - , -S(0~o ~CH2 -<~, ~ , ~, =.
R1_N N-- Rt_N
' or where Z is 0, NR8, or S; and R8 is defined as R1 above;
R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring ,system containing 0, 1, 2, 3, or 4 heteroatom.s selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more group.s selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, C1 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl-5 alkyl, aminoCl S alkyl, hydroxycarbonyl, hydroxycarbonylCl-s alkyl, or hydroxycarbonylC l 5 alkoxy, W O 9712625a PCTrUS97/00572 -(CH2)n-aryl, wherein n=1-4 and aryl i~ defined as a five or six membered mono or polycyclic aromatic ring system cont:~;ning 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or sub~tituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, Cl 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl 5 alkyl, aminoC~ 5 alkyl, hydroxycarbonyl, hydroxycarbonylC1 5 alkyl, or hydroxycarbonylC1 ~ alkoxy, l 0 halogen, hydroxyl, C1 5alkylcarbonylamino, arylC1 5 alkoxy, Cl 5 alkoxycarbonyl, aminocarbonyl, C1 5 alkylaminocarbonyl, C1 5 alkylcarbonyloxy, C3 ~s cycloalkyl, oxo, amino, C l 3 alkylamino, aminoC l 3 alkyl, arylaminocarbonyl, arylC1 salkylaminocarbonyl, aminocarbonyl, aminocarbonyl-C 1 4 alkyl, hydroxycarbonyl, hydroxycarbonyl Cl 5 alkyl, C1 6alkyl, either un~ubstituted or substituted, with one or more groups selected from halogen, hydroxyl, C1 5alkylcarbonylamino, arylC~ 5 alkoxy, C1 5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, Cl 5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, W O 97/26250 PCTrUS97/00572 Cl 3 alkylamino, aminoCl 3 alkyl, arylaminocarbonyl, arylCl 5alkylaminocarbonyl, aminocarbonyl, aminocarbonyl-CI 4 alkyl, hydroxycarbonyl, or hydroxycarbonyl Cl 5 alkyl, provided that the carbon atom to which R3 and R4 are attached bear.s only one heteroatom, -(CH2~m C-CH, -(CH2)mC_C-Cl 6alkyl, -(CH2)m C-C-C3 7cycloalkyl, -(CH2)m C_C- aryl, -(CH2)m C-C-Cl 6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C 1-6 alkyl, -(CH2)m CH=CH-C3 7cycloalkyl, -(CH2)m CH=CH aryl, -(CH2)m CH-C~ C1 6 alkyl aryl, -(CH2)m SO2C1 6 alkyl, or -(CH2)m SO2Cl 6 alkylaryl;
20 R5 i~
hydrogen, fluorine, C1 8 allcyl, hydroxyl, hydroxy C 1 6 alkyl, carboxy, carboxy C1 6 alkyl, C 1 6 alkyloxy.
C3-8 cycloalkyl, aryl C1 6 alkyloxy, aryl, aryl C1 6 alkyl, C 1 6 alkylcarbonyloxy, amino, W O 97/26250 PCTrUS97/00572 amino C1 6 alkyl, Cl-6 alkylamino, C1 6 alkylamino C1 6 alkyl, aryl amino, aryl amino C 1-6 alkyl, aryl C1 6 alkylamino, aryl C1 6 alkylamino (~1-6 aLkyl, aryl carbonyloxy, aryl C1 6 alkylcarbonylo~y, 11) ~1-6 dialkylamino, C1 6 dialkylamino C1 6 alkyl, C1 6 alkylaminocarbonyloxy, (~1-8 alkylsulfonylamino, C 1 -P~ alkylsulfonylamino C 1-6 alkyl, aryl .sulfonylamino C1 6 alkyl, aryl C1 6 alkylsulfonylamino, aryl C1 6 alkylsulfonylamino C 1 6 alkyl, C1 8 alkyloxycarbonylamino, Cl ~s alkyloxycarbonylamino Cl g alkyl, aryl oxycarbonylamino C1 8 alkyl, aryl C1 2 alkyloxycarbonylamino, aryl C1 8 alkyloxycarbonylamino Cl ~ alkyl, Cl ~ alkylcarbonylamino, C1 8 alkylcarbonylamino C1 6 alkyl, aryl carbonylamino C1 6 alkyl, aryl C1 6 alkylcarbonylamino, aryl C1 6 alkylcarbonylamino C1 6 alkyl, aminocarbonylamino C1 6 alkyl, Cl-tg alkylaminocarbonylamino, C1 8 alkylaminocarbonylamino C1 6 aLkyl, aryl aminocarbonylamino C1 6 alkyl, aryl C1 8 alkylaminocarbonylamino, aryl Cl g alkylaminocarbonylamino C1 6 aLkyl, aminosulfonylamino C1 6 alkyl, W 097/26250 PCT~US97/00572 C l ~ alkylaminosulfonylamino, Cl ~ alkylaminosulfonylamino Cl-6 alkyl, aryl aminosulfonylamino C1 6 alkyl, aryl C 1 8 alkylaminosulfonylamino, aryl Cl ~ alkylaminosulfonylamino C1 6 alkyl, C 1 6 alkylsulfonyl, C 1-6 alkylsul~onyl C 1 -6alkyl, aryl sulfonyl C1 6alkyl~
aryl C 1 6 alkyl~sulfonyl, 1~) aryl C1 6 alkylsulfonyl C l 6alkyl, C 1 6 alkylcarbonyl, C 1 6 alkylcarbonyl C1 6 alkyl, aryl carbonyl C1 6alkyl, aryl C1 6 alkylcarbonyl, aryl C1 6 alkylcarbonyl C1 6alkyl, C1 6 aLkylthiocarbonylamino, C1 6 alkylthiocarbonylamino C1 6 alkyl, aryl thiocarbonylamino C1 6 alkyl, aryl C1 6 alkylthiocarbonylamino, 2~) aryl C1 6 alkylthiocarbonylamino C1 6 alkyl, aminocarbonyl C1 6 alkyl, or Cl ~ alkylaminocarbonyl, or Cl g alkylaminocarbonyl C1 6 alkyl, or aryl aminocarbonyl C1 6 alkyl, aryl C1 8 alkylaminocarbonyl, aryl C1 8 alkylaminocarbonyl C1 6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R ~ and R2; and 30 R6 and R7 are independently hydrogen, '-Cl ~ alkyl, aryl, aryl C l ~¢ alkyl, W O 97/26250 PCTrUS97/00572 hydroxy, C 1 8 alkyloxy, aryloxy, aryl Cl-6 alkyloxy, ~1-8 alkylcarbonyloxy Cl 4 alkyloxy, aryl Cl ~ alkylcarbonyloxy Cl 4 alkyloxy, Cl g alkylaminocarbonylmethyleneoxy, or C1 8 dialkylaminocarbonylmethyleneoxy where m and n are integers 0-6.
In a class of compounds of the invention are compounds of the formula:
o O R4 Il 11 1 X~(CH2)m~Y~(CH2)n~C~ IN-cH2-c-NH-cH-cH-co2R6 R3 R~
and pharmaceutically acceptable salts thereof, wherein W 097126250 PCT~US97/00572 X is R ~ N~5 R ~ N~N--R1 N~N ~ R1 N~
N~R Z i ~3 ~' ~ , ~ . or wherein n is 2-4, and n' is 2 or 3, and wherein Rl and R2 are independently selected from the group con~isting of hydrogen, F, Cl, Br, I, C 1 - 10 alkyl, C~3-8 cycloalkyl~
aryl, aryl C1 ~ alkyl, PCT~US97/00572 amino, amino C1 8 alkyl, C l 3 aeylamino, C 1-3 acylamino C l -g alkyl, Cl -6 alkylamino, C1 6 alkylamino C1 ~ alkyl, C 1 6 dialkylamino, C1 6 dialkylamino Cl ~ alkyl, C l 4 alkoxy, C1 4 alkoxy Cl 6 alkyl, earboxy, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, C l 3 alkoxyearbonyl C l 6 alkyl, carboxy Cl 6 alkyloxy and hydroxy, hydroxy Cl 6 alkyl;
R5 i~
hydrogen, fluorine, C l 8 alkyl, hydroxyl, hydroxy Cl 6 alkyl, carboxy, earboxy C1 6 alkyl, C1 6 alkyloxy.
C3-8 eyeloalkyl, aryl C1 6 alkyloxy, aryl, aryl Cl 6 alkyl, C l 6 alkylcarbonyloxy, amino, C1 6 alkylamino, W O 97/26250 PCT~US97/00572 amino Cl 6 alkyl, C1 6 alkylamino C1 6 alkyl, aryl amino, aryl amino C1 6 alkyl, aryl C1 6 alkylamino, aryl Cl-6 alkylamino C1 6 alkyl, aryl carbonyloxy, aryl C1 6 alkylcarbonyloxy, C 1 6 dialkylamino, C 1 6 dialkylamino C 1 6 alkyl, C1 6 alkylaminocarbonyloxy, Cl ~ alkylsulfonylamino, C l p~ alkylsulfonylamino C 1 6 alkyl, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkyl~ulfonylamino, aryl C1 6 alkylsulfonylamino C1 6 alkyl, C l ~3 alkyloxycarbonylamino, Cl ~, alkyloxycarbonylamino C~-8 alkyl, aryl C1 8 alkyloxycarbonylamino, aryl oxycarbonylamino, aryl oxycarbonylamino C1 8 alkyl, aryl C1 8 alkyloxycarbonylamino Cl ~¢ alkyl, Cl ~ alkylcarbonylamino, Cl-8 alkylcarbonylamino C1 6 alkyl, aryl carbonylamino C1 6 alkyl, aryl carbonylamino, aryl C1 6 aL~cylcarbonylamino, aryl Cl-6 alkylcarbonylamino C1 6 alkyl, C 1-~ alkylaminocarbonylamino, aminocarbonylamino, aminocarbonylamino C1 6 alkyl, C1 8 alkylaminocarbonylamino C1 6 alkyl, aryl aminocarbonylamino C1 6 alkyl, W O 97/2625n PCT~US97/00572 aryl aminocarbonylamino, aryl Cl 8 alkylaminocarbonylamino, aryl C1 g alkylaminocarbonylamino Cl 6 alkyl, aminosulfonylamino Cl 6 alkyl, S aminosulfonylamino, C l ~ alkylaminosulfonylamino, C l ~ alkylamino~ulfonylamino C l 6 alkyl, aryl aminosulfonylamino C1 6 alkyl, aryl aminosulfonylamino, l O aryl C l -8 alkylaminosul~onylamino, aryl C1 8 alkylaminosulfonylamino Cl 6 alkyl, C 1 6 alkylsulfonyl, C1 6 alkylsulfonyl Cl 6alkyl, aryl sulfonyl, aryl sulfonyl Cl -6aLkyl, aryl alkylsulfonyl, aryl C1 6 alkylsulfonyl, aryl Cl 6 alkylsulfonyl Cl 6alkyl~
C 1 6 alkylcarbonyl, Cl 6 alkylcarbonyl C1 6 alkyl, aryl carbonyl Cl 6alkyl, aryl carbonyl, aryl Cl 6 alkylcarbonyl, aryl Cl-6 alkylcarbonyl C1 6alkyl, C l -6 alkylthiocarbonylamino, Cl 6 alkylthiocarbonylamino Cl 6 alkyl, aryl thiocarbonylamino C l 6 alkyl, aryl thiocarbonylamino, aryl C1 6 alkylthiocarbonylamino, aryl C1 6 alkylthiocarbonylamino Cl 6 alkyl, ~ aminocarbonyl C1 6 alkyl, aminocarbonyl, Cl ~ alkylaminocarbonyl, Cl g alkylaminocarbonyl Cl 6 alkyl, ,~
W 097/26250 PCT~US97/00572 aryl aminocarbonyl C1 6 alkyl, aryl aminocarbonyl, aryl C l g allcylaminocarbonyl, aryl C 1 ~ alkylaminocarbonyl C 1 6 alkyl, S wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R l and R2; and R6 and R7 are independently hydrogen, C 1-~ alkyl, aryl Cl 8 alkyl, hydroxy, C l ~s alkyloxy, aryl, aryl C1 6 alkyloxy, Cl ~ alkylcarbonyloxy Cl 4 alkyloxy, aryl Cl g alkylcarbonyloxy C1 4 alkyloxy, C1 8 alkylaminocarbonylmethyleneoxy, or Cl g diaLkylaminocarbonylmethyleneoxy, 20 where m and n are integers 0-6.
In a subclass of the class of compounds described above are compounds of the formula O o R4 Il 11 1 X~(CH2)m~Y~(CH2)n~C~NI -cH2-c-NH-cH-cH-co2R6 R3 Rs 25 and pharmaceutically acceptable salts thereof, wherein xi~;
W O 97/2625~ PCTAUS97/00572 R~N~S R~N N--Rl N~N ~ R1N3~
(C~l~ R1~, ~ N
s~~ ~ ~J' H2N ~/ , or N~
wherein n' is 2 or 3, and S wherein R I and R2 are independently selected from the group con.sisting of hydrogen, F, Cl, Br, I, C ~ alkyl, C3-8 cycloalkyl, aryl, aryl Cl ~s alkyl, amino, amino C1 g alkyl, C 1 3 acylamino, C 1-3 acylamino C 1 ~ alkyl, C 1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, C1 6 dialkylamino, S C 1 6 dialkylamino Cl ~s alkyl, C 1 4 alkoxy, Cl 4 alkoxy C1 6 alkyl, carboxy, carboxy C1 6 alkyl, Cl 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy C1 6 alkyloxy and hydroxy, hydroxy C 1 6 alkyl;
Y is o NHR6 R8 ~(CH2)0-6~, -C-C-, -C=C~, - C-, -C- ~ -N -, -O-, R80 o R8 - SO2NH-, - NHSO2-, -N-C-, -C--N-, ~ , R 1_ N N-- R 1_ N
where Z is 0, NR8, or S; and R8 is defined as R1 above;
W O 97/262~0 PCTAUS97/00572 R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, Cl 5 alkylcarbonyloxy, C1 5 alkoxycarbonyl, C1 S alkyl, aminoC1 5 alkyl, hydroxycarbonyl, hydroxycarbonylCl 5 alkyl, or hydroxycarbonylCl 5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl i,~; defined a,~ a five or six membered mono or polycyclic aromatic ring system cont~ining 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, C1 5 alkylcarbonyloxy, C1 5 alkoxycarbonyl, Cl 5 alkyl, aminoCl 5 alkyl, hydroxycarbonyl, hydroxycarbonylCl 5 alkyl, or hydroxycarbonylCl 5 alkoxy, halogen, hydroxyl, C1 5alkylcarbonylamino, arylCI 5 alkoxy, Cl 5 alkoxycarbonyl, aminocarbonyl, Cl 5 alkylaminocarbonyl, Cl 5 alkylcarbonyloxy, C3 ~s cycloalkyl, oxo, amino, C~ 3 alkylamino, aminoC l 3 alkyl, arylaminocarbonyl, arylC1 5alkylaminocarbonyl, arninocarbonyl-C l 4 alkyl, hydroxycarbonyl, hydroxycarbonyl Cl S alkyl, Cl 6alkyl, either un,~ubstituted or substituted, with one or more S groups selected from halogen, hydroxyl, C1 5alkylcarbonylamino, arylCl 5 alkoxy, Cl 5 alkoxycarbonyl, aminocarbonyl, C1 5 alkylaminocarbonyl, C1 5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1 3 alkylamino, aminoC1 3 alkyl, arylaminocarbonyl, arylC1 5alkylaminocarbonyl, aminocarbonyl-C1 4 alkyl, hydroxycarbonyl, or hydroxycarbonyl C1 5 alkyl, provided that the carbon atom to which R3 and R4 are attached bears only one 1 5 heteroatom, -(CH2)m C_CH, -(CH2)m C_C-C1 6 alkyl, -(CH2)m C-C-C3 7cycloalkyl, -(CH23m C-C- aryl, -(CH2)m C_C-C1 6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C1 6 alkyl, -(CH2)m CH=CH-C3 7cycloalkyl, -(CH2)m CH=CH aryl, 2~ -(CH2)m CH=CH C1-6 alkyl aryl, -(CH2)mSO2C I 6alkyl~ or -(CH2)mSO2C 1 6 alkylaryl, R5 is hydrogen, fluorine, C 1 8 alkyl, hydroxyl, hydroxy C1 6 alkyl, W O 97/2625~ PCTnUS97100572 carboxy, carboxy C1 6 alkyl, C 1 6 alkylo~y.
C3 ~ cycloalkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyl, C 1 6 alkylcarbonyloxy, amino C1 6 alkyl, amino, (~1-6 alkylamino, C1 6 alkylamino C1 6 alkyl, aryl amino C1 6 alkyl, aryl amino, aryl C1 6 alkylarnino, aryl C1 6 alkylamino C1 6 alkyl, aryl, aryl C1 6 alkylcarbonyloxy, C 1 6 dialkylamino, C 1 6 dialkylamino C 1 6 alkyl, Cl -6 alkylaminocarbonyloxy, C 1 8 alkylsulfonylamino, C1 8 alkylsulfonylamino C1 6 alkyl, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkylsulfonylarnino, aryl C1 6 alkylsulfonylamino C1 6 alkyl, Cl ~s alkyloxycarbonylamino, C 1 ~ alkyloxycarbonylamino C l ~ alkyl, aryl oxycarbonylamino C~ ~ alkyl, aryl oxycarbonylamino, aryl C1 8 alkyloxycarbonylamino, aryl C1 8 alkyloxycarbonylamino C1 8 alkyl, C1 8 alkylcarbonylamino, C1 8 alkylcarbonylamino C1 6 alkyl, W O 97/26250 PCT~US97/OOS72 aryl carbonylamino C 1 6 alkyl, aryl carbonylamino, aryl C1 6 alkylcarbonylamino, aryl C1 6 alkylcarbonylamino Cl-6 alkyl, S aminocarbonylamino C1 6 alkyl, aminocarbonylamino, C l ~s alkylaminocarbonylamino, Cl g alkylaminocarbonylamino C1 6 alkyl, aryl aminocarbonylamino C1 6 alkyl, aryl aminocarbonylamino, aryl Cl-~s alkylaminocarbonylamino, aryl Cl p~ alkylaminocarbonylamino C1 6 aL~yl, aminosulfonylamino C 1 6 alkyl, aminosulfonylamino, l S C 1-~ alkylaminosulfonylamino, C1 ~ alkylaminosul~onylamino C1 6 alkyl, aryl aminosulfonylamino C1 6 alkyl, aryl aminosulfonylamino, aryl C l ~s alkylaminosulfonylamino, aryl C 1 8 alkylaminosulfonylamino C 1 6 alkyl, C 1 6 alkylsulfonyl, C1 6 alkylsulfonyl C1 6alkyl, aryl sulfonyl C 1 6alkyl, aryl sulfonyl, aryl C 1 6 alkylsulfonyl, aryl C1 6 alkylsulfonyl Cl 6alkyl, C1 6 alkylcarbonyl, C1 6 alkylcarbonyl C1 6 alkyl, aryl carbonyl C1 6alkyl, aryl carbonyl, aryl C1 6 alkylcarbonyl, aryl C 1 6 alkylcarbonyl C l 6alkyl, C 1 6 alkylthiocarbonylamino, C1 6 alkylthiocarbonylamino C1 6 alkyl, W O 97/26250 PC~US97/00572 aryl thiocarbonylamino C1 6 alkyl, aryl thiocarbonylamino, aryl C1 6 alkylthiocarbonylamino, aryl C1 6 alkylthiocarbonylamino C1 6 alkyl, S aminocarbonyl Cl -6 alkyl, aminocarbonyl, C 1 8 alkylaminocarbonyl, C 1-8 alkylaminocarbonyl C 1-6 alkyl, aryl aminocarbonyl C1 ~ alkyl, aryl aminocarbonyl, aryl Cl ~ allcylaminocarbonyl, or aryl C1 8 allcylaminocarbony} C1 6 alkyl, wherein alkyl groups and aryl groups may be un~ubstituted or substituted with one or more substituents selected from R 1 and R2; and R6 ~cmd R7 are independently hydrogen, Cl ~ alkyl, aryl, aryl C1 8 alkyl, hydroxy, Cl p~ alkyloxy, aryl oxy, aryl Cl-6 alkyloxy, C1 8 alkylcarbonyloxy Cl 4 alkyloxy, aryl C1 8 alkylcarbonyloxy Cl 4 alkyloxy, Cl ~s aL~cylaminocarbonylmethyleneoxy, or C1 8 dialkylaminocarbonylmethyleneoxy, where m and n are integers 0-6.
In a group of the subclass are compounds having the formula ~.
ll R4 X-(CH2)m-Y-(CH2)n-C-IN-CH2-C-NH-CH-ICH-CO2R6 R3 R~
and pharmaceutically acceptable salts thereof, wherein X i,s RZ R~
R1~-~ Rl~ N R2~N' R~ N ~\
S~ ' ~ .
H2N ~/ , or N~/
wherein Rl and R2 are independently selected from the group consisting of hydrogen or ammo, amino Cl ~ allcyl;
W O 97/26250 PCT~USg7/OOS72 J
Y lS
-(CH2)0-4- , - C-N - , -O-, or -N-Rg is hydrogen or aryl Co g alkyl;
R3 i.~
S hydrogen, a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, Cl 5 aLkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl 5 alkyl, aminoCl 5 aLkyl, hydroxycarbonyl, hydroxycarbonylC1 5 alkyl, or hydroxycarbonylC l -S
alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl i~ defined as a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C 1-3 alkoxy, C 1-5 alkylcarbonyloxy, C 1-5 alkoxycarbonyl, C1 5 alkyl, aminoC1 5 alkyl, hydroxycarbonyl, hydroxycarbonylC 1 -S alkyl, or hydroxycarbonylC l -S
alkoxy, C3-8 cycloalkyl, or C 1 6alkyl, either unsubstituted or substituted, with C3-8 cycloalkyl;
R4 i~
hydrogen, -(CH2)n-aryl, wherein n=0-4 and aryl is defined as a six - membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups - selected from hydroxyl, halogen, cyano, trifluoromethyl, C1 3 alkoxy, C1 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, C1 5 alkyl, aminoCl 5 alkyl, hydroxycarbonylCo 5 alkyl, or hydroxycarbonylC 1-5 alkoxy, W O 97/26250 PCT~US97/00572 C1 6alkyl, or -(CH2)0-4 C_CH;
~5 i,~
hydrogen, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkylsulfonylamino, aryl C 1 6 alkylsulfonylamino C1 6 alkyl, Cl -~ alkylsulfonylamino, Cl ~ alkylsulfonylamino C1 6 alkyl, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkylsulfonylamino, aryl Cl-6 alkylsulfonylamino C1 6 alkyl, aminosulfonylamino Cl G alkyl, aminosulfonylamino, Cl ~ alkylaminosulfonylamino, C 1 ~ alkylaminosulfonylamino C 1 6 alkyl, aryl aminosul~onylamino C 1-6 alkyl, aryl aminosulfonylamino, aryl C l ~ alkylaminosulfonylaminol aryl C1 ~ alkylaminosulfonylamino C1 6 alkyl, C 1 6 alkylsulfonyl, C1 6 alkylsulfonyl Cl 6alkyl, aryl sulfonyl C 1 6alkyl, aryl sulfonyl, aryl Cl 6 alkylsulfonyl, aryl C1 6 alkylsulfonyl Cl 6alkyl, wherein alkyl groups and aryl groups may be un.substituted or substituted with one or more substituents selected from R1 and R2;
W O 97/262~0 PCTrUS97100572 R6 is hydrogen, Cl ~ alkyl, or aryl, S aryl Cl 8 alkyl;
m is an integer selected from O to 6; and n is an integer selected from O to 6.
In a subgroup of the group are compounds having the formula 1~l 1~l 14 X~(CH2)m~Y~(CH2)n~C~IN-CH2-C-NH-CH-ICH-CO2R6 and pharmaceutically acceptable salt.s thereof, wherein X is S~-- ~/ ~/ H2N
H~l N ~ J ,~
N~N~ ~,~ or W 097/26250 PCT~US97/00572 - 2~ -Y is -(CH2)0 4-, 0 -N- , or -N- ;
R3 is hydrogen, S methyl, , or R4 is hydrogen, 1 0 methyl, 2 ~
~' ~
=CH;
W O 97/262~0 PCTAUS97/00572 , - 29 -.,R5 i,s hydrogen, or S R6 j~
hydrogen, methyl, ethyl, or t-butyl;
m i.s an integer se3ected from 0 to 6; and n i,s an integer selected from 0 to 6.
Specific examples of this subgroup include 1 5 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-alanine t-butyl ester 4-(2-~minothiazol-4-yl)butanoyl-glycyl-2(S~-phenylsulfonamido-~(3-alanine 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl)-,13-alanine methyl ester 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl~-,13-alanine 25 trifluoroacetate salt 5 -(2-Pyridylamino)pentanoylglycyl -2(S)-phenylsulfonamido-,13 -alanine ethyl ester 30 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine trifluoroacetate ~alt W 097/26250 PCTrUS97/OOS72 4-(2-Bocamino-pyridin-6-yl)butanoyl-~arcosine-3(R)-[(2-indol-3-yl)ethyl]-~3-alanine ethyl e,~ter 4-(2-Aminopyridin-6-yl)butanoyl-~sarcosine-3(R)-[(2-indol-3-yl)ethyl] -5 ,B-alanine 4-(2-Boc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsul~onamido-~-alanine t-butyl ester 10 4-(2-Aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,B-alanine 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-13-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-sarcosine-3(~ 2-(indol-3-yl)ethyl]-~-alanine 4-(2-Bocamino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3 (R)-(2-phenethyl)-~-alanine ethyl ester 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-~-alanine ethyl ester hydrochloride 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R~-(2-25 phenethyl)-,~-alanine 4-(2-Boc-amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol -3 -yl)ethyl] -,B-alanine 30 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-~(2-indol-3-yl)ethyl] -,13-alanine 4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-,(3-alanine ethyl ester W O 97126250 PCT~US97/00572 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-,B-alanine ethyl ester 4-(2-~mino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~3-S alanine 4-(Pyridin-4-yl)butanoyl-N-(2-phenylethyl)glycyl-3(R)-(2-phenethyl)-,13-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-13-alanine 4-(2-~3OC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-,B-alanine benzyl ester 4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-~-alanine 4-(2-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-20 ~-alanine 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanine ethyl ester 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-~3-alanir~e 3 -[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-,B-alanine ethyl ester 3-~(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-~3-alanine W O 97/26250 PCT~US97/00572 N- ~ N'-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl ~ -3(R)-rnethyl-,B-alanine benzyl ester N-[N'-~3-( 1 -Piperazinyl)l~enzoyl]glycyl]-3(R)-methyl-~-alanine 5 trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-,(3-alanine methyl ester 10 N-~N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-,B-alanine trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]-N'-(2-phenethyl)glycyl]-3~R)-(2-phenethyl)-13-alanine methyl ester 1~
N-[N'-[3-( 1 -Piperazinyl)benzoyl]-N -(2-phenethyl)glycyl] -3(R)-(2-phenethyl)-13-alanine trifluoroacetic acid salt 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanol-glycyl-~-a~anine 20 t-butyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)but~noyl-glycyl-~-alanine 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-pyridin-3-yl-~3-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)pyridin-3-yl-~-alanine Ethyl N-pyridin-4-ylisonipecotyl-N-cyclopropylglycine-3(S)-ethynyl-,~-alanine W O 97/26250 PCTrUS97/00572 N-Py~idin-ylisonipecotyl-N-cyclopropylglycine-3 (S)-ethynyl-~-alanine Ethyl N-pyridin-4-ylnipecotyl-N-cyclopropylglycine-3(S)-ethynyl-~-S alanine N-Pyridin-4-ylnipecotyl-N-cyclopropylglycine-3(S)-ethynyl-,~-alanine 10 4-(1,~',3,4-Tetrahydro- 1 ,~-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)gly-3(S)ethynyl-~B-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3 (S)-ethynyl-~-alanine, 3- ~ 2-~5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino]-acetylamino } -3(S)-pyridin-3-yl- propionic acid.
and pharmaceutically acceptable salts, such as the trifluoroacetate salt 20 and the hydrochloric acid salt Compounds of the invention are ak;o useful for inhibiting the bone resorption activity of mammalian osteoclast cells by a~1mini~tering a pharmacologically effective amount of the compound to a patient in need of such activity to inhibit the activity of mammalian 25 osteoclasts.
Compounds of the invention are also useful for inhibiting tumor growth in m~mmals. Pharrnacologically effective amounts of the compounds, including pharamaceutically acceptable salts thereof, are administered to the m~mmal, to inhibit tumor growth. The growth of 30 tumors depends on an adequate blood supply, which in turn depends on growl:h of new vessels into the tumor. New vessels are stimulated by factors ,secreted by the tumor. Inhibition of angiogenesis can cause tumor regres.sion in animals.
W O 97/26250 PCT~US97/00572 Compounds of the invention are also useful for treating and preventing diabetic retinopathy in m;~mm~ls. Pharmacologically effective amount,~ of the compounds, including pharamaceutically acceptable salts thereof, are ~lministered to the mammal, to inhibit diabetic re~inopathy.
Compounds are also useful in the prevention of restenosis of vessels.
The te~n "bone resorption activity" means the process by which osteoclast~ solubilize bone minerals and increase the activity of 10 en~ymes that degrade bone matrix.
Compounds of the invention are u,seful for inhibiting the binding of fibrinogen to blood platelets and for inhibiting the aggregation of blood platelets. The above-mentioned compounds can be used in a method of acting upon a fibrinogen receptor which comprises 15 administering a therapeutically effective but non-toxic amount of such compound to a m~mm~l, preferably a human. A pha~naceutical composition comprising a pharmaceutically acceptable carrier and, dispersed therein, an effective but non-toxic amount of such compound i~; another feature of this invention.
The invention also includes the u~;e of a compound of the invention, or a phalmaceutically acceptable .salt thereof, in the manufacture of a medicament for inhibiting the aggregation of blood platelets, preventing platelet thrombosis, preventing thromboembolism or preventing reocclu~ion, in a m~mm~l.
~ETAIL~D DESCRIPTION OF THE INVENTION
Fibrinogen receptor antagonist compounds of Formula I
are useful in a method of inhibiting the binding of fibrinogen to blood platelets and for inhibiting the aggregation of blood platelet~.
30 Fibrinogen receptor antagonists of this invention are illu~trated by compounds having the folmula: , The following compoullds were tested and found to inhibit platelet aggregation with IC50 values between about 0.01 ~M and 100 ,uM.
W 097/262S0 PCTrUS97/00572 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,13-alanine t-butyl ester 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-S alanine 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-~-alanine methyl ester 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-~2-phenethyl)-~-alanine trifluoroacetate ~alt 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenyl~ulfonamido-~-alanine ethyl ester 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine trifluoroacetate salt 4-(2-Bocamino-pyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-20 yl)ethyl]-~-alanine ethyl ester 4-(2-Aminopyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-yl)ethyl] -,B-alanine 4-(2-Boc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,~3-alanine t-butyl ester 4-(2-Aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-13-alanine 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-~2-(indol-3-yl)ethyl]-~-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-~3-alanine W O 97/262~0 PCT~US97/00572 4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-,B-alanine ethyl ester 4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3(R)-(2-~S phenethyl)-,B-~lanine ethy~ e~ter hydrochloride 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-13-alanine 1 0 4-(2-Boc-amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol -3-yl)ethyl] -~-alanine 4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3 (R)-~(2-indol-3 -yl)ethyl] -,~-alanine 4-(2-Bocamino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3(R)-methyl-~-alanine ethyl ester 4-(2-Amino-pyridin-6-yl)butanoyl -N-cyclopropylglycyl-3 (R)-methyl-,(3-20 alanine ethyl e~ter 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3 (R)-methyl-~-alanine 25 4-(Pyridin-4-yl)butanoyl-N-(2-phenylethyl)glycyl-3(R~-(2-phenethyl)-~-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-~-alanine 4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3 (R)-methyl-~-alanine benzyl ester . CA 02242877 1998-07-13 W O 97/2625~ PCT~US97/OOS72 4-(2-lBOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-- methyl-~3-alanine 4-(2-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3 (R)-methyl-S ~-alanine 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanine ethyl ester 1 0 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,13-alanine 3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarco.sine-3(R)-(2-phenethyl)-~-alanine ethyl ester 3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-,(3-alanine N- { N '-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl } -3(R)-20 methyl-,13-alanine benzyl ester N-[N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-methyl-}3-alanine trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl ester N-[N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-~B-alanine trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]-N'-(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl ester - 3~ -N-[N'-L3-( 1 -Piperazinyl)benzoyl]-N -(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine trifluoroacetic acid salt 4-(1,2,3 ,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)butanol-glycyl-,(3-alanine 5 t-butyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)butanoyl-glycyl-,B-alanine 10 4-(1 ,2 ,3 ,4-Tetrahydro- 1 ,X -naphthyridin -7 -yl)butanoyl -glycyl -3 (S )- pyridin-~-yl-,~-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,g-naphthyridin-7-yl)butanoyl-glycyl-3(S)pyridin-3-yl-~(3-alnine Ethyl N-pyridin-4-yli~onipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-alanine N-Pyridin-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-20 alanine Ethyl N-pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~3-alanine 25 N-Pyridin-4-ylnipecotyl -N -cyclopropylglycyl-3 (S )-ethynyl -~13 -alanine 4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3(S)ethynyl-,B-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,~s-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3 (S)-ethynyl-,B-alanine .
W O 97/26250 PCTrUS97/00572 One test which is used to evaluate fibrinogen receptor antagonist activity i,s based on evaluation of inhibition of ADP-stimulated platelets. Aggregation requires that fibrinogen bind to and occupy the platelet fibrinogen receptor site. Inhibitors of fibrinogen 5 binding inhibit aggregation. In the ADP-stimulated platelet aggregation assa~ u,~ed to determine inhibition associated with the compounds claimed in the instant invention, human platelets are isolated from fresh blood, collected into acid citrate/dextrose by differential centrifugation follo~wed by gel filtration on Sepharose 2B in divalent ion-free Tyrode's 1() buffer (pH 7.4) containing 2% bovine serum albumin.
Platelet aggregation is measured at 37~C in a Chronolog aggregometer. The reaction mixture contains gel-filtered human platelets (2 x 10P~ per ml~, fibrinogen ~100 micrograms per ml (ug/ml)), Ca2t (1 mM), and the compound to be tested. The aggregation is 15 initiated by adding 10 mM ADP 1 minute after the other components are added. The reaction is then allowed to proceed for at least 2 minutes. The extent of inhibition of aggregation is expressed as the percentage of the rate of aggregation observed in the absence of inhibitor. The I~50 is the dose of a particular compound inhibiting 20 aggregation by 50% relative to a control lacking the compound.
The terrn "pharmaceutically acceptable salts" shall mean non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid. Representative salts include the following salts:
25 acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clav~lanate, citrate, dihydrochloride, edetate, edisylate, estolate, e,sylate, fumarate, gluceptate, gluconate, glllt~m~te, glycollylarsanilate, hexyLresorcinate, hydrabamine, hydrobromide, hydrochloride, 30 hydroxynapthoate, iodide, isothionate, lactate7 lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote, palmitate, panthothenate, phosphate/diphosphate, polygalacturonate, W O 97/26250 PCTrUS97100572 salicylate, stearate, ,~ubacetate, ~uccinate, tannate, tartrate, teoclate, tosylate, triethiodide, trifluoroacetate, ;md valerate.
Compounds of the present invention are chiral; included within the ,~cope of the pre~ent invention are racemic mixtures and separated enantiomers of the general formula. Furthermore, all dia~tereomers, including E, Z isomers, of the general formula are included in the pre~ent scope. Furthermore, hydrates as well as anhydrous compositions and polymorphs of the general formula are within the present inventioIl.
Prodrugs, ,~;uch a,~; ester derivatives of described compounds, are compound derivatives which, when absorbed into the bloodstream of a warrn-blooded ~nim;~l, cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.
lS The term "pharmaceutically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the biological or medical respon,se of a tissue, system or animal that is being ,sought by a researcher or clinician. The term "anti-coagulant" shall include heparin, and warfarin. The term "thrombolytic agent" shall 20 include agents such as streptokina,se and tissue pla,sminogen activator.
The term "platelet anti-aggregation agent" ~hall include agents such as aspirin and dipyridamole.
The term "alkyl" means straight or branched alkane containing 1 to about 10 carbon atoms, e.g., methyl, ethyl, n-propyl, 25 isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexy, octyl radicals and the like, straight or branched alkene containing 2 to about 10 carbon atoms, e.g., propylenyl, buten-l-yl, isobutenyl, pentenylen- I -yl, 2~2-methylbuten- 1 -yl, 3-methylbuten- 1 -yl, hexen- I -yl, hepten-l-yl, and octen-1-yl radicals and the like, or straight or branched 30 alkyne cont~ining 2 to about 10 carbon atoms, e.g.~ ethynyl, propynyl, butyn-l-yl, butyn-2-yl, pentyn-l-yl, pentyn-2-yl, 3-methylbutyn-1-yl, hexyn-l-yl, hexyn-2-yl, hexyn-3-yl, 3,3-dimethylbutyn-1-yl radical,~; and the like.
The term "aryl" means a 5- or 6-membered aromatic ring c conta-;ning 0, 1, or 2 heteroatoms selected from O, N, and S. Examples of aryl include phenyl, pyridine, pyrimidine, imidazole, thiophene, oxazole, isoxazole, thiazole, and amino- and halogen- substituted derivatives thereof.
The te~n~ "alkyloxy" or "alkoxy" include an alkyl portion where alkyl is as defined above. Examples of alkoxy include methyloxy, propyloxy, and butyloxy.
The term.s "arylalkyl" and "alkylaryl" include an alkyl portion where alkyl is as defined above and to include an aryl portion whe~ aryl i~ as defined above. The Co-n or (~l-n designation where n may be an integer from 1-10 or 2-10 respectively refer~ to the alkyl component of the arylalkyl or alkylaryl unit. Examples of arylalkyl include benzyl, fluorobenzyl, chlorobenzyl, phenylethyl, phenylpropyl, fluorophenylethyl, chlorophenylethyl, thienylmethyl, thienylethyl, and thienylpropyl. Examples of alkylaryl include toluene, ethylbenzene, propylbenzene, methylpyridine, ethylpyridine, propylpyridine, hutylpyridine, butenylpy~idine, and pentenylpyridine.
The term "halogen" includes fluorine, chlorine, iodine and bromine.
The term "oxy" means an oxygen (O) atom. The term "thio" means a sulfur (S) atom. Under standard nonmenclature used throughout this disclosure, the terminal portion of the designated side chain is described first followed by the adjacent functionality toward the point of attachment. For example, a C1 6 alkyl substituted with C1 5 alkyl-carbonylamino i.s e4uivalent to HO
C1 6-alkyl-N-C-C1 s-alkyl Compounds of the invention where X i~s a 5-membered monocyclic aromatic ring system, e.g., a thiazole system, can be prepared by forming an alkyl ester stituted derivative of the ring, 35 e.g., methyl 4-(2-aminothiazol-4-yl)butanoate, forming the corresponding acid with HCl, and reacting with an amine to form the final product.
Compounds of the invention where X is a 6-membered monocyclic aromatic ring system, e.g., a pyridine Isystem, can be 5 prepared using 2-aminopyridine, 2-aminopicoline, 4-vinyl pyridine, etc., as described in Schemes 3, 4, and 10.
Compounds of the invention where X is a 9-membered polycyclic aromatic fused ring system can be prepared by reacting a substituted 5-membered ring starting material such as 2-amino-3-bromo 10 thiophene, 2-nitro-3-bromo thiophene, 2-amino-3-bromo pyrrole, and 2-amino-3-bromo furan, with an appropriate compound under suitable ring clo,sure conditions to effect formation of the 9-membered fused ring system.
Compounds of the invention where X is a 10-membered 15 polycyclic aromatic ring sy,stem can be prepared using a starting material such a~s naphthyridin (Hamada, Y. et al., Chem. Pha7 m. Bull.
Soc., 1971, 19(9), 1857-1862), or by reacting an aminoaldehyde pyridine with a .suitable ketone under suitable ring closure conditions to effect formation of the 10-membered fused ring system.
The examples illustrate procedures for preparing compounds of the invention where Y is -(CH2)0 4, -O-~ and -N(R~)-.
To make compounds where Y is -N(R~)C(O)-, an acid such as compound 1-4 can be subjected to a Curtius reaction to form the amine, and subsequent condensation to give the final product.
In the schemes and examples below, various reagent symbols have the following meanings:
BOC
(or Boc): t-butyloxycarbonyl Pd-C: Palladium on activated carbon catalyst DMF: Dimethylformamide DMSO: Dimethyl,sulfoxide CBZ: Carbobenzyloxy CH2Cl2: Methylene chloride W 097/2625(~ PCT~US97/00572 CHC13: chloroform r EtOH: ethanol MeOH: methanol EtOAc: ethyl acetate S HOAc: acetic acid BOP. Benzotriazol- l -yloxytris(dimethylamino)phosphonium, hexafluorophosphate EDC: 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride Oxone: potassium peroxymonosulfate LDA: Lithium diisopropylamide PYC]LU: Chloro-N,N,N',N'-bis(pentamethylene)formamidinium hexafluorophosphate l~ The compounds of the present invention can be ~1mimistered in such oral forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granule~., elixirs, tinctures, suspensions, syrups, and emulsions.
Like~ise, they may be ~(lmini~tered in intravenous (bolus or infusion), intra~)eritoneal, subcutaneous, or intramusculsar form, all using forms well Iknown to those of ordinary skill in the pharmaceutical arts. An effec~ive but non-toxic amount of the compound desired can be employed as an anti-aggregation agent.
Compounds of the invention may be administered to patients where prevention of thrombosis by inhibiting binding of fibrinogen to the platelet membrane glycoprotein complex IIb/IIIa receptor is desired. They are useful in surgery on peripheral arteries (arteIial grafts, carotid endarterectomy) and in cardiovascular surgery where manipulation of arteries and organs, and/or the interaction of r 3Q plate]ets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelets may form thrombi and thromboemboli. Compounds of this invention may be ~lmini~tered to these surgical patients to prevent the formation of thrombi and thromboemboli.
W O 97/26250 PCT~US97100572 Extracorporeal circulation is routinely used for cardiovascular surgery in order to o~ygenate blood. Platelets adhere to surfaces of the extracorporeal circuit. Adhesion is dependent on the interaction between gp rIb/IIIa on the platelet membrane,~ and 5 fibrinogen adsorbed to the surface of the circuit. (Gluszko et al., Ame7 .
~1. Physiol., 252(H), 615-621 (1987)). Platelets released from artificial surfaces show impaired hemostatic function. Compounds of the invention may be admini,stered to prevent adhesion.
Other applications of these compounds include prevention 10 of platelet thrombosis, thromboembolism and reocclusion during and after thrombolytic therapy and prevention of platelet thrombosis, thromboembolism and reocclusion after angioplasty or coronary artery bypass procedures. They may also be used to prevent myocardial infarction.
The dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of a-lministration; the renal and hepatic function of the patient; and the particular compound 20 or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug re~luired to prevent, counter, or arrest the progress of the condition.
Oral dosages of the present invention, when used for the 2~ indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 100 mg/kg/day and preferably 0.01-SQ mg/kg/day and more preferably 0.01-20 mg/kg/day, e.g. 0.1 mg/kg/day, 1.0 mg/kg/day, 5.0 mg/kg/day, or 10 mg/lcg/day.
Intravenously, the most preferred doses will range from about 1 to 30 about 10 mg/kg/minute during a constant rate infusion.
Advantageously, compounds of the present invention may be administered in divided doses of two, three, or four times daily.
Furthermore, preferred compounds for the present invention can be ~ministered in intranasal form via topical use of suitable intranasal W O 97/26250 PCTrUS97/00572 vehicles, or via transdermal routes, using those forms of transderrnal skin patches well known to those of ordinary skill in that art. To be a~lmini~stered in the form of a transdermal delivery system, the dosage ~lministration will, or course, be continuous rather that intermittent 5 throughout the dosage regime.
In the methods of the present invention, the compounds hereiIl described in detail can form the active ingredient, and are typically a(lmini~tered in admixture with suitable pharrnaceutical diluents, e~cipients or carriers (collectively referred to herein as 10 "carrier" materials) suitably selected with respect to the intended form of ~lministration, that is, oral tablets, capsules, elixirs, syrups and the like, cmd consistent with convention pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, 15 non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert 20 carrier such as ethanol, glycerol, water and the like. ~oreover, when desired or necessary, suitable binder,s, lubricants, distintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lacto~e, corn-sweeteners, natural and synthetic gums such as acacia, 25 tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms inclucle sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators inclucle, without limitation, starch methyl cellulose, agar, bentonite, 30 xanthan gum and the like.
., The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar ve,sicles, large unilamellar vesicles and multilamellar W 097126250 PCTrUS97/00572 vesicles. Liposomes can be forrned from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
Compounds of the present invention may also be delivered by the u,se of monoclonal antibodies as individual carriers to which the ;S compound molecule~ are coupled. The compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copo}ymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxy-ethyl-aspartamide-phenol, or polyethyleneoxide-polylysine sub,stituted 10 with palmitoyl residue~s. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled relea,se of a drug, for example, polylactic acid~
polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, 1~ polyacetals, polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block copolymers of hydrogels.
The compounds of the present invention can also be co-administered with suitable anticoagulation agents or thrombolytic agents such a~ plasminogen activators or streptokinase to achieve synergistic 20 effect.~ in the treatment of various vascular pathologie.s. They may also be combined with heparin, aspirin, or warfarin.
The novel compounds of the pre.sent invention were prepared according to the procedure of the following examples. The most preferred compounds of the invention are any or all of those 25 specifically set forth in these examples. These compounds are not, however, to be construed as forming the only genus that is considered as the invention, and any combination of the compounds or their moieties may itself form a genus. The following examples further illustrate detail,s for the preparation of the compounds of the present invention.
3(~ Tho.se skilled in the art will readily understand that known variations of the condition.s and processes of the following preparative procedures can be used to prepare these compounds. All temperatures are degrees Cel.siu.s unless otherwise noted.
W O 97/26250 PCT~US97/00572 SCHEME I
~ ~i CO2H HCI-H2N~H SO
Br2, MeOH
0~C to RT Boc-Gly-OH
r NMM, isobutyl chloroformate, Br~~/~CO2Me O EtOAc, -t5~Cto RT
0 1-2 BocHN~J~ N~CO2-t-Bu ,, EtOH, ~ H H NH--S~2 H~N~<N~ CO2Me TFA, CH2ci2 1-3 -1 5~C
INTEGRIN l~ECEPTOR ANTAGONISTS
BACKGROUND OF THE INVEN~ION
S The invention relates generally to modulating cell adhe,c;ionand to inhibiting the binding of fibrinogen and other proteins to blood plate~ets, and inhibiting the aggregation of blood platelets specifically to the gp IIb/IIIa fibrinogen receptor ,site. Fibrinogen is a glycoprotein present in blood plasma that participates in platelet aggregation and in fibrin formation. Platelets are cell-like anucleated *agments, found in the blood of al} m~mm~ls, that al.~io participate in blood coagulation.
Inter~action o~ fibrinogen with the IIb/~Ila receptor site is known to be essential for normal platelet function.
When a blood vessel is damaged by an injury or other causative factor, platelets adhere to the disrupted subendothethial surface. The adherent platelets subse4uently release biologically active constituents and aggregate. Aggregation is initiated by the binding of agonists, such as thrombin, epinephrine, or ADP to specific platelet membrane receptors. Stimulation by agonists results in exposure of latent fibrinogen receptors on the platelet surface, and binding of fibrinogen to the glycoprotein IIb/IIIa receptor complex.
Attempts have been made to use natural product~; and synthetic peptides to determine the mechanism of adhesion and platelet aggregation. For example, Rouslahti and Pierschbacher in Sci~nce, 238, 491-497 (1987), describe adhesive proteins such as fibronectin, vitronectin, osteopontin, collagens, thrombospondin, fibrinogen, and von Willebrand factor that are present in extracellular matrices and in blood. The protein.s contain the tripeptide arginine-glycine-aspartic acid (RGD) as their glycoprotein IIb/IIIa recognition site. These arginine-glycine-aspartic acid cont~ining tripeptides are recognized by at least one member of a family of structurally related receptors, integrins, which are heterodimeric proteins with two membrane-spanning subunits. The authors state that the conformation of the tripeptide sequence in the individual proteins may be critical to recognition ~pecificity.
Cheresh in Proc. Nat'l Acad. Sci. U.S.A., 84, 6471-6475, "
(19~7), describes an Arg-Gly-Asp directed adhesion receptor expressed by human endothethial cells that is structurally similar to the rIb/IIIa complex on platelet,s but is antigenically and functionally distinct. This receptor is directly involved in endothelial cell attachment to fibrinogen, von Willebrand factor, and vitronectin.
Pier.schbacher and Rouslahti, in J. of Biol. Chem., 262, (36), 17294- 1729~i (19~7) hypothesized that the Arg-Gly-Asp sequence alone would be a sufficient signal for receptor recognition and binding and that, therefore, the conforrnation of the tri-peptide sequence would be determinative. Various synthetic peptides were produced and the authors concluded that the sterochemical con~orrnation of ~rg-Gly-Asp as influenced by enantiomeric substitutions or additions to this ,sequence signi~lcantly influenced receptor-ligand interaction. The authors furlher showed that cyclization of a decapeptide by forming a disulfide bridge between non-terminal residues Pen and Cys, rendered the peptide much less effective at inhibiting attachment to fibronectin.
In Proc. Nat'l Acad. Sci. U.S.A., 81, 59g5-59~8 (19~4), the same author.s describe tetrapeptide variants of the cell recognition site of fibronectin that retain attachment-promoting activity. Peptides having a tetrapeptide recognition site are described in U.S. Pat. Nos. 4,5~9,g81 and 4,614,517. A number of large polypeptide fragments in the cell-binding domain of fibronectin have cell-attachment activity. For example, see U.S. Pat. Nos. 4,517,686, 4,661,111 and U.S. Pat. No.
4,57~,079.
Ruggeri et al., Proc. Nat'l Acad. Sci. U.S.A., 83, 570~s-5712 (19~S6) explore a serie~s of synthetie peptides designed in lengths to 16 residues, that contain ~GD and a valine attached to the aspartic acid residue of RGD that inhibit fibrinogen binding to platelets. See also Koc~ewiak et al., Biochem., 23, 1767-1774 ~19~4); Ginsberg et al., WO 97/26250 PCTrUS97/00572 J. Bivl. Ch~m., 260(7), 3931-3936 (1985); and Haverstick et al., Bloocl, ~6(4), 946-952 (1985). Other inhibitor~ are disclosed in Eur. Pat. App.
Nos. 275,74g and 298,820.
A number of low molecular weight polypeptide factors 5 have been i,solated from ~nake venom. These ~actors apparently have high affinity for ~e gp ~Ib/~IIa complex. For example, Huang et al., J.
B~ol C~em., 262, 16157-16163 (1987); Huang et al., Bic~che~7list)y, 28, 661-1566 (1989) describe the primary structure of the venom trigramin which is a 72 amino acid polypeptide that contains the RGD subunit.
10 Echixtatin is another compound which has high affinity for the gp IIb/Il[Ia complex. This polypeptide contains 49 amino acids and has the RGI~l subunit and various disulfide bridges. Gan et al., J. Biol. Chem., 263, 19827-19g32 (1988). See also, Dennis ~t al., Proc. Nat'l Acad. Sci.
USA. 87, 2471-2475 (1989). However, the,~ie snake venom factor.~ also 15 have high affinity for other members of the adhesive protein receptor famihy including the vitronectin and fibronectin receptors so are not selective for the gp IIb/IIIa complex.
While it is known that the tripeptide sequence Arg-Gly-Asp i,s present in certain polypeptides that can duplicate or inhibi~ the cell 20 attachment-promoting effect~ of fibronectin and vitronectin, the tripeptide Arg-Gly-Asp has low activity. At present, there is Iittle understanding of how other amino acid~ coupled to this .~equence influence binding specificity. U.S. Pat. No 5,023,233, assigned to Merck & Co., Inc., discloses small cyclic hexapeptides which contain the 25 sequence Arg-Gly-Asp and are useful platelet aggregation inhibitors.
U.S. Pat. No. 5,037,808 discloses the use of indolyl platelet-aggregation inhibitorx which are believed to act by antagonizing interactions between ~ibrinogen and/or extracellular matrix proteins and the platelet gp IIb/llIa receptor. U.S. Pat. No. 5,037,~08 disclol~;es guanidino peptide 30 mimetic compounds that retain an Asp residue which inhibit platelet aggregation. The application PCT/US90/02746 describes the use of antibody-poly-peptide conjugates wherein said polypeptides contain the Arg-Gly-Asp (RGD) sequence.
W O 97/26250 PCT~US97/00572 The application PCT/US91/00564 discloses the use of large cyclic peptides containing RGD flanked by proline residues which are platelet aggregation inhibitors. The application PCT/US90/03788 discloses small cyclic platelet aggregation inhibitors which are synthetic 5 cyclic pentapeptides containing the tripeptide sequence Arg-Gly-Asp and a thioether linkage in the cycle. The application PCT/US90/05367 published May 2, 1991 also discloses the use of peptides and pseudopeptides such as N-amidino-piperidine-3-carboxylglycyl-L-aspartyl-L-valine that inhibit platelet aggregation and thrombus 10 formation in m~mmalian blood. The application Eur. Pat. App. No.
91103462.7 discloses linear compounds which can include internal piperazinyl or piperidinyl derivatives. Eur. Pat. App. No. 91300179.~, assigned to Merck & Co., Inc., and published on July 17, 1991 discloses linear polypeptide fibrinogen receptor antagonists. Eur. Pat. App. No.
1~ 90101404.3 discloses compounds of the Rl-A-(W)a-X-(CH2)b-(Y)c-B-Z-COOR wherein Rl is a guandidino or amidino moiety and A and B
are chosen from specific monosubstituted aryl or heterocyclic moieties.
While a multitude of compounds or peptide analogs believed to inhibit platelet aggregation by inhibiting binding to a blood 20 platelet by fibrinogen are known, the present invention provides novel fibrinogen receptor antagonists that have significant binding activity and are, therefore, useful for the reasons stated herein. A number of very serious diseases and disorders involve hyperthrombotic complications which lead to intravascular thrombi and emboli. Myocardial infarction, 2~ stroke, phlebitis and a number of other serious conditions create the need for novel and effective fibrinogen receptor antagonists.
SIIMMARY OF THE INVENTION
The invention is a compound of the formula:
1~l ~ R4 X~(CH2)m~Y~(CH2)n~C~N~CH2~C~NH~CH-CH-CO2R6 3() 1 3 R~
W O 97/262~0 PCT~US97/00572 and pharmaceutically acceptable salts thereof, wherein xi,s a 5- or 6-membered monocyclic aromatic ring ~ystem containing 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either S unsubstituted or substituted with R l or R2, or a 9- to 10-membered polycyclic ring ~ystem, wherein one or more of the ring~ is aromatic, cont~ning 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either un.~ubstituted or substituted with R l or R2, wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, Cl -10 alkyl, C3-8 cycloalkyl, aryl, aryl Cl ~s alkyl, amino, amino C l ~ alkyl, C1 3 acylamino, C1 3 acylamino C1 8 alkyl, C 1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, 2~ C 1-6 dialkylamino, C1 6 dialkylamino Cl ~ alkyl, C 1 4 alkoxy, CI 4 alkoxy C1 6 alkyl, carboxy, carboxy C1 6 alkyl, Cl 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy C1 6 alkyloxy and W O 97/26250 PCT~US97/00572 hydroxy, hydroxy C1 6 alkyl;
Y i.s O N H R6 Rs -(CH2)0 6- . -C-C-, -C=C~, - C -, - C - ~ -N - , -0-, - S02NH - , - NHS02 -, -N-~- -C ~N - , -S(0~o ~CH2 -<~, ~ , ~, =.
R1_N N-- Rt_N
' or where Z is 0, NR8, or S; and R8 is defined as R1 above;
R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring ,system containing 0, 1, 2, 3, or 4 heteroatom.s selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more group.s selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, C1 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl-5 alkyl, aminoCl S alkyl, hydroxycarbonyl, hydroxycarbonylCl-s alkyl, or hydroxycarbonylC l 5 alkoxy, W O 9712625a PCTrUS97/00572 -(CH2)n-aryl, wherein n=1-4 and aryl i~ defined as a five or six membered mono or polycyclic aromatic ring system cont:~;ning 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or sub~tituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, Cl 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl 5 alkyl, aminoC~ 5 alkyl, hydroxycarbonyl, hydroxycarbonylC1 5 alkyl, or hydroxycarbonylC1 ~ alkoxy, l 0 halogen, hydroxyl, C1 5alkylcarbonylamino, arylC1 5 alkoxy, Cl 5 alkoxycarbonyl, aminocarbonyl, C1 5 alkylaminocarbonyl, C1 5 alkylcarbonyloxy, C3 ~s cycloalkyl, oxo, amino, C l 3 alkylamino, aminoC l 3 alkyl, arylaminocarbonyl, arylC1 salkylaminocarbonyl, aminocarbonyl, aminocarbonyl-C 1 4 alkyl, hydroxycarbonyl, hydroxycarbonyl Cl 5 alkyl, C1 6alkyl, either un~ubstituted or substituted, with one or more groups selected from halogen, hydroxyl, C1 5alkylcarbonylamino, arylC~ 5 alkoxy, C1 5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, Cl 5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, W O 97/26250 PCTrUS97/00572 Cl 3 alkylamino, aminoCl 3 alkyl, arylaminocarbonyl, arylCl 5alkylaminocarbonyl, aminocarbonyl, aminocarbonyl-CI 4 alkyl, hydroxycarbonyl, or hydroxycarbonyl Cl 5 alkyl, provided that the carbon atom to which R3 and R4 are attached bear.s only one heteroatom, -(CH2~m C-CH, -(CH2)mC_C-Cl 6alkyl, -(CH2)m C-C-C3 7cycloalkyl, -(CH2)m C_C- aryl, -(CH2)m C-C-Cl 6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C 1-6 alkyl, -(CH2)m CH=CH-C3 7cycloalkyl, -(CH2)m CH=CH aryl, -(CH2)m CH-C~ C1 6 alkyl aryl, -(CH2)m SO2C1 6 alkyl, or -(CH2)m SO2Cl 6 alkylaryl;
20 R5 i~
hydrogen, fluorine, C1 8 allcyl, hydroxyl, hydroxy C 1 6 alkyl, carboxy, carboxy C1 6 alkyl, C 1 6 alkyloxy.
C3-8 cycloalkyl, aryl C1 6 alkyloxy, aryl, aryl C1 6 alkyl, C 1 6 alkylcarbonyloxy, amino, W O 97/26250 PCTrUS97/00572 amino C1 6 alkyl, Cl-6 alkylamino, C1 6 alkylamino C1 6 alkyl, aryl amino, aryl amino C 1-6 alkyl, aryl C1 6 alkylamino, aryl C1 6 alkylamino (~1-6 aLkyl, aryl carbonyloxy, aryl C1 6 alkylcarbonylo~y, 11) ~1-6 dialkylamino, C1 6 dialkylamino C1 6 alkyl, C1 6 alkylaminocarbonyloxy, (~1-8 alkylsulfonylamino, C 1 -P~ alkylsulfonylamino C 1-6 alkyl, aryl .sulfonylamino C1 6 alkyl, aryl C1 6 alkylsulfonylamino, aryl C1 6 alkylsulfonylamino C 1 6 alkyl, C1 8 alkyloxycarbonylamino, Cl ~s alkyloxycarbonylamino Cl g alkyl, aryl oxycarbonylamino C1 8 alkyl, aryl C1 2 alkyloxycarbonylamino, aryl C1 8 alkyloxycarbonylamino Cl ~ alkyl, Cl ~ alkylcarbonylamino, C1 8 alkylcarbonylamino C1 6 alkyl, aryl carbonylamino C1 6 alkyl, aryl C1 6 alkylcarbonylamino, aryl C1 6 alkylcarbonylamino C1 6 alkyl, aminocarbonylamino C1 6 alkyl, Cl-tg alkylaminocarbonylamino, C1 8 alkylaminocarbonylamino C1 6 aLkyl, aryl aminocarbonylamino C1 6 alkyl, aryl C1 8 alkylaminocarbonylamino, aryl Cl g alkylaminocarbonylamino C1 6 aLkyl, aminosulfonylamino C1 6 alkyl, W 097/26250 PCT~US97/00572 C l ~ alkylaminosulfonylamino, Cl ~ alkylaminosulfonylamino Cl-6 alkyl, aryl aminosulfonylamino C1 6 alkyl, aryl C 1 8 alkylaminosulfonylamino, aryl Cl ~ alkylaminosulfonylamino C1 6 alkyl, C 1 6 alkylsulfonyl, C 1-6 alkylsul~onyl C 1 -6alkyl, aryl sulfonyl C1 6alkyl~
aryl C 1 6 alkyl~sulfonyl, 1~) aryl C1 6 alkylsulfonyl C l 6alkyl, C 1 6 alkylcarbonyl, C 1 6 alkylcarbonyl C1 6 alkyl, aryl carbonyl C1 6alkyl, aryl C1 6 alkylcarbonyl, aryl C1 6 alkylcarbonyl C1 6alkyl, C1 6 aLkylthiocarbonylamino, C1 6 alkylthiocarbonylamino C1 6 alkyl, aryl thiocarbonylamino C1 6 alkyl, aryl C1 6 alkylthiocarbonylamino, 2~) aryl C1 6 alkylthiocarbonylamino C1 6 alkyl, aminocarbonyl C1 6 alkyl, or Cl ~ alkylaminocarbonyl, or Cl g alkylaminocarbonyl C1 6 alkyl, or aryl aminocarbonyl C1 6 alkyl, aryl C1 8 alkylaminocarbonyl, aryl C1 8 alkylaminocarbonyl C1 6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R ~ and R2; and 30 R6 and R7 are independently hydrogen, '-Cl ~ alkyl, aryl, aryl C l ~¢ alkyl, W O 97/26250 PCTrUS97/00572 hydroxy, C 1 8 alkyloxy, aryloxy, aryl Cl-6 alkyloxy, ~1-8 alkylcarbonyloxy Cl 4 alkyloxy, aryl Cl ~ alkylcarbonyloxy Cl 4 alkyloxy, Cl g alkylaminocarbonylmethyleneoxy, or C1 8 dialkylaminocarbonylmethyleneoxy where m and n are integers 0-6.
In a class of compounds of the invention are compounds of the formula:
o O R4 Il 11 1 X~(CH2)m~Y~(CH2)n~C~ IN-cH2-c-NH-cH-cH-co2R6 R3 R~
and pharmaceutically acceptable salts thereof, wherein W 097126250 PCT~US97/00572 X is R ~ N~5 R ~ N~N--R1 N~N ~ R1 N~
N~R Z i ~3 ~' ~ , ~ . or wherein n is 2-4, and n' is 2 or 3, and wherein Rl and R2 are independently selected from the group con~isting of hydrogen, F, Cl, Br, I, C 1 - 10 alkyl, C~3-8 cycloalkyl~
aryl, aryl C1 ~ alkyl, PCT~US97/00572 amino, amino C1 8 alkyl, C l 3 aeylamino, C 1-3 acylamino C l -g alkyl, Cl -6 alkylamino, C1 6 alkylamino C1 ~ alkyl, C 1 6 dialkylamino, C1 6 dialkylamino Cl ~ alkyl, C l 4 alkoxy, C1 4 alkoxy Cl 6 alkyl, earboxy, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, C l 3 alkoxyearbonyl C l 6 alkyl, carboxy Cl 6 alkyloxy and hydroxy, hydroxy Cl 6 alkyl;
R5 i~
hydrogen, fluorine, C l 8 alkyl, hydroxyl, hydroxy Cl 6 alkyl, carboxy, earboxy C1 6 alkyl, C1 6 alkyloxy.
C3-8 eyeloalkyl, aryl C1 6 alkyloxy, aryl, aryl Cl 6 alkyl, C l 6 alkylcarbonyloxy, amino, C1 6 alkylamino, W O 97/26250 PCT~US97/00572 amino Cl 6 alkyl, C1 6 alkylamino C1 6 alkyl, aryl amino, aryl amino C1 6 alkyl, aryl C1 6 alkylamino, aryl Cl-6 alkylamino C1 6 alkyl, aryl carbonyloxy, aryl C1 6 alkylcarbonyloxy, C 1 6 dialkylamino, C 1 6 dialkylamino C 1 6 alkyl, C1 6 alkylaminocarbonyloxy, Cl ~ alkylsulfonylamino, C l p~ alkylsulfonylamino C 1 6 alkyl, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkyl~ulfonylamino, aryl C1 6 alkylsulfonylamino C1 6 alkyl, C l ~3 alkyloxycarbonylamino, Cl ~, alkyloxycarbonylamino C~-8 alkyl, aryl C1 8 alkyloxycarbonylamino, aryl oxycarbonylamino, aryl oxycarbonylamino C1 8 alkyl, aryl C1 8 alkyloxycarbonylamino Cl ~¢ alkyl, Cl ~ alkylcarbonylamino, Cl-8 alkylcarbonylamino C1 6 alkyl, aryl carbonylamino C1 6 alkyl, aryl carbonylamino, aryl C1 6 aL~cylcarbonylamino, aryl Cl-6 alkylcarbonylamino C1 6 alkyl, C 1-~ alkylaminocarbonylamino, aminocarbonylamino, aminocarbonylamino C1 6 alkyl, C1 8 alkylaminocarbonylamino C1 6 alkyl, aryl aminocarbonylamino C1 6 alkyl, W O 97/2625n PCT~US97/00572 aryl aminocarbonylamino, aryl Cl 8 alkylaminocarbonylamino, aryl C1 g alkylaminocarbonylamino Cl 6 alkyl, aminosulfonylamino Cl 6 alkyl, S aminosulfonylamino, C l ~ alkylaminosulfonylamino, C l ~ alkylamino~ulfonylamino C l 6 alkyl, aryl aminosulfonylamino C1 6 alkyl, aryl aminosulfonylamino, l O aryl C l -8 alkylaminosul~onylamino, aryl C1 8 alkylaminosulfonylamino Cl 6 alkyl, C 1 6 alkylsulfonyl, C1 6 alkylsulfonyl Cl 6alkyl, aryl sulfonyl, aryl sulfonyl Cl -6aLkyl, aryl alkylsulfonyl, aryl C1 6 alkylsulfonyl, aryl Cl 6 alkylsulfonyl Cl 6alkyl~
C 1 6 alkylcarbonyl, Cl 6 alkylcarbonyl C1 6 alkyl, aryl carbonyl Cl 6alkyl, aryl carbonyl, aryl Cl 6 alkylcarbonyl, aryl Cl-6 alkylcarbonyl C1 6alkyl, C l -6 alkylthiocarbonylamino, Cl 6 alkylthiocarbonylamino Cl 6 alkyl, aryl thiocarbonylamino C l 6 alkyl, aryl thiocarbonylamino, aryl C1 6 alkylthiocarbonylamino, aryl C1 6 alkylthiocarbonylamino Cl 6 alkyl, ~ aminocarbonyl C1 6 alkyl, aminocarbonyl, Cl ~ alkylaminocarbonyl, Cl g alkylaminocarbonyl Cl 6 alkyl, ,~
W 097/26250 PCT~US97/00572 aryl aminocarbonyl C1 6 alkyl, aryl aminocarbonyl, aryl C l g allcylaminocarbonyl, aryl C 1 ~ alkylaminocarbonyl C 1 6 alkyl, S wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R l and R2; and R6 and R7 are independently hydrogen, C 1-~ alkyl, aryl Cl 8 alkyl, hydroxy, C l ~s alkyloxy, aryl, aryl C1 6 alkyloxy, Cl ~ alkylcarbonyloxy Cl 4 alkyloxy, aryl Cl g alkylcarbonyloxy C1 4 alkyloxy, C1 8 alkylaminocarbonylmethyleneoxy, or Cl g diaLkylaminocarbonylmethyleneoxy, 20 where m and n are integers 0-6.
In a subclass of the class of compounds described above are compounds of the formula O o R4 Il 11 1 X~(CH2)m~Y~(CH2)n~C~NI -cH2-c-NH-cH-cH-co2R6 R3 Rs 25 and pharmaceutically acceptable salts thereof, wherein xi~;
W O 97/2625~ PCTAUS97/00572 R~N~S R~N N--Rl N~N ~ R1N3~
(C~l~ R1~, ~ N
s~~ ~ ~J' H2N ~/ , or N~
wherein n' is 2 or 3, and S wherein R I and R2 are independently selected from the group con.sisting of hydrogen, F, Cl, Br, I, C ~ alkyl, C3-8 cycloalkyl, aryl, aryl Cl ~s alkyl, amino, amino C1 g alkyl, C 1 3 acylamino, C 1-3 acylamino C 1 ~ alkyl, C 1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, C1 6 dialkylamino, S C 1 6 dialkylamino Cl ~s alkyl, C 1 4 alkoxy, Cl 4 alkoxy C1 6 alkyl, carboxy, carboxy C1 6 alkyl, Cl 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy C1 6 alkyloxy and hydroxy, hydroxy C 1 6 alkyl;
Y is o NHR6 R8 ~(CH2)0-6~, -C-C-, -C=C~, - C-, -C- ~ -N -, -O-, R80 o R8 - SO2NH-, - NHSO2-, -N-C-, -C--N-, ~ , R 1_ N N-- R 1_ N
where Z is 0, NR8, or S; and R8 is defined as R1 above;
W O 97/262~0 PCTAUS97/00572 R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, Cl 5 alkylcarbonyloxy, C1 5 alkoxycarbonyl, C1 S alkyl, aminoC1 5 alkyl, hydroxycarbonyl, hydroxycarbonylCl 5 alkyl, or hydroxycarbonylCl 5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl i,~; defined a,~ a five or six membered mono or polycyclic aromatic ring system cont~ining 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, C1 5 alkylcarbonyloxy, C1 5 alkoxycarbonyl, Cl 5 alkyl, aminoCl 5 alkyl, hydroxycarbonyl, hydroxycarbonylCl 5 alkyl, or hydroxycarbonylCl 5 alkoxy, halogen, hydroxyl, C1 5alkylcarbonylamino, arylCI 5 alkoxy, Cl 5 alkoxycarbonyl, aminocarbonyl, Cl 5 alkylaminocarbonyl, Cl 5 alkylcarbonyloxy, C3 ~s cycloalkyl, oxo, amino, C~ 3 alkylamino, aminoC l 3 alkyl, arylaminocarbonyl, arylC1 5alkylaminocarbonyl, arninocarbonyl-C l 4 alkyl, hydroxycarbonyl, hydroxycarbonyl Cl S alkyl, Cl 6alkyl, either un,~ubstituted or substituted, with one or more S groups selected from halogen, hydroxyl, C1 5alkylcarbonylamino, arylCl 5 alkoxy, Cl 5 alkoxycarbonyl, aminocarbonyl, C1 5 alkylaminocarbonyl, C1 5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1 3 alkylamino, aminoC1 3 alkyl, arylaminocarbonyl, arylC1 5alkylaminocarbonyl, aminocarbonyl-C1 4 alkyl, hydroxycarbonyl, or hydroxycarbonyl C1 5 alkyl, provided that the carbon atom to which R3 and R4 are attached bears only one 1 5 heteroatom, -(CH2)m C_CH, -(CH2)m C_C-C1 6 alkyl, -(CH2)m C-C-C3 7cycloalkyl, -(CH23m C-C- aryl, -(CH2)m C_C-C1 6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C1 6 alkyl, -(CH2)m CH=CH-C3 7cycloalkyl, -(CH2)m CH=CH aryl, 2~ -(CH2)m CH=CH C1-6 alkyl aryl, -(CH2)mSO2C I 6alkyl~ or -(CH2)mSO2C 1 6 alkylaryl, R5 is hydrogen, fluorine, C 1 8 alkyl, hydroxyl, hydroxy C1 6 alkyl, W O 97/2625~ PCTnUS97100572 carboxy, carboxy C1 6 alkyl, C 1 6 alkylo~y.
C3 ~ cycloalkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyl, C 1 6 alkylcarbonyloxy, amino C1 6 alkyl, amino, (~1-6 alkylamino, C1 6 alkylamino C1 6 alkyl, aryl amino C1 6 alkyl, aryl amino, aryl C1 6 alkylarnino, aryl C1 6 alkylamino C1 6 alkyl, aryl, aryl C1 6 alkylcarbonyloxy, C 1 6 dialkylamino, C 1 6 dialkylamino C 1 6 alkyl, Cl -6 alkylaminocarbonyloxy, C 1 8 alkylsulfonylamino, C1 8 alkylsulfonylamino C1 6 alkyl, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkylsulfonylarnino, aryl C1 6 alkylsulfonylamino C1 6 alkyl, Cl ~s alkyloxycarbonylamino, C 1 ~ alkyloxycarbonylamino C l ~ alkyl, aryl oxycarbonylamino C~ ~ alkyl, aryl oxycarbonylamino, aryl C1 8 alkyloxycarbonylamino, aryl C1 8 alkyloxycarbonylamino C1 8 alkyl, C1 8 alkylcarbonylamino, C1 8 alkylcarbonylamino C1 6 alkyl, W O 97/26250 PCT~US97/OOS72 aryl carbonylamino C 1 6 alkyl, aryl carbonylamino, aryl C1 6 alkylcarbonylamino, aryl C1 6 alkylcarbonylamino Cl-6 alkyl, S aminocarbonylamino C1 6 alkyl, aminocarbonylamino, C l ~s alkylaminocarbonylamino, Cl g alkylaminocarbonylamino C1 6 alkyl, aryl aminocarbonylamino C1 6 alkyl, aryl aminocarbonylamino, aryl Cl-~s alkylaminocarbonylamino, aryl Cl p~ alkylaminocarbonylamino C1 6 aL~yl, aminosulfonylamino C 1 6 alkyl, aminosulfonylamino, l S C 1-~ alkylaminosulfonylamino, C1 ~ alkylaminosul~onylamino C1 6 alkyl, aryl aminosulfonylamino C1 6 alkyl, aryl aminosulfonylamino, aryl C l ~s alkylaminosulfonylamino, aryl C 1 8 alkylaminosulfonylamino C 1 6 alkyl, C 1 6 alkylsulfonyl, C1 6 alkylsulfonyl C1 6alkyl, aryl sulfonyl C 1 6alkyl, aryl sulfonyl, aryl C 1 6 alkylsulfonyl, aryl C1 6 alkylsulfonyl Cl 6alkyl, C1 6 alkylcarbonyl, C1 6 alkylcarbonyl C1 6 alkyl, aryl carbonyl C1 6alkyl, aryl carbonyl, aryl C1 6 alkylcarbonyl, aryl C 1 6 alkylcarbonyl C l 6alkyl, C 1 6 alkylthiocarbonylamino, C1 6 alkylthiocarbonylamino C1 6 alkyl, W O 97/26250 PC~US97/00572 aryl thiocarbonylamino C1 6 alkyl, aryl thiocarbonylamino, aryl C1 6 alkylthiocarbonylamino, aryl C1 6 alkylthiocarbonylamino C1 6 alkyl, S aminocarbonyl Cl -6 alkyl, aminocarbonyl, C 1 8 alkylaminocarbonyl, C 1-8 alkylaminocarbonyl C 1-6 alkyl, aryl aminocarbonyl C1 ~ alkyl, aryl aminocarbonyl, aryl Cl ~ allcylaminocarbonyl, or aryl C1 8 allcylaminocarbony} C1 6 alkyl, wherein alkyl groups and aryl groups may be un~ubstituted or substituted with one or more substituents selected from R 1 and R2; and R6 ~cmd R7 are independently hydrogen, Cl ~ alkyl, aryl, aryl C1 8 alkyl, hydroxy, Cl p~ alkyloxy, aryl oxy, aryl Cl-6 alkyloxy, C1 8 alkylcarbonyloxy Cl 4 alkyloxy, aryl C1 8 alkylcarbonyloxy Cl 4 alkyloxy, Cl ~s aL~cylaminocarbonylmethyleneoxy, or C1 8 dialkylaminocarbonylmethyleneoxy, where m and n are integers 0-6.
In a group of the subclass are compounds having the formula ~.
ll R4 X-(CH2)m-Y-(CH2)n-C-IN-CH2-C-NH-CH-ICH-CO2R6 R3 R~
and pharmaceutically acceptable salts thereof, wherein X i,s RZ R~
R1~-~ Rl~ N R2~N' R~ N ~\
S~ ' ~ .
H2N ~/ , or N~/
wherein Rl and R2 are independently selected from the group consisting of hydrogen or ammo, amino Cl ~ allcyl;
W O 97/26250 PCT~USg7/OOS72 J
Y lS
-(CH2)0-4- , - C-N - , -O-, or -N-Rg is hydrogen or aryl Co g alkyl;
R3 i.~
S hydrogen, a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy, Cl 5 aLkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl 5 alkyl, aminoCl 5 aLkyl, hydroxycarbonyl, hydroxycarbonylC1 5 alkyl, or hydroxycarbonylC l -S
alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl i~ defined as a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C 1-3 alkoxy, C 1-5 alkylcarbonyloxy, C 1-5 alkoxycarbonyl, C1 5 alkyl, aminoC1 5 alkyl, hydroxycarbonyl, hydroxycarbonylC 1 -S alkyl, or hydroxycarbonylC l -S
alkoxy, C3-8 cycloalkyl, or C 1 6alkyl, either unsubstituted or substituted, with C3-8 cycloalkyl;
R4 i~
hydrogen, -(CH2)n-aryl, wherein n=0-4 and aryl is defined as a six - membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups - selected from hydroxyl, halogen, cyano, trifluoromethyl, C1 3 alkoxy, C1 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, C1 5 alkyl, aminoCl 5 alkyl, hydroxycarbonylCo 5 alkyl, or hydroxycarbonylC 1-5 alkoxy, W O 97/26250 PCT~US97/00572 C1 6alkyl, or -(CH2)0-4 C_CH;
~5 i,~
hydrogen, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkylsulfonylamino, aryl C 1 6 alkylsulfonylamino C1 6 alkyl, Cl -~ alkylsulfonylamino, Cl ~ alkylsulfonylamino C1 6 alkyl, aryl sulfonylamino C1 6 alkyl, aryl sulfonylamino, aryl C1 6 alkylsulfonylamino, aryl Cl-6 alkylsulfonylamino C1 6 alkyl, aminosulfonylamino Cl G alkyl, aminosulfonylamino, Cl ~ alkylaminosulfonylamino, C 1 ~ alkylaminosulfonylamino C 1 6 alkyl, aryl aminosul~onylamino C 1-6 alkyl, aryl aminosulfonylamino, aryl C l ~ alkylaminosulfonylaminol aryl C1 ~ alkylaminosulfonylamino C1 6 alkyl, C 1 6 alkylsulfonyl, C1 6 alkylsulfonyl Cl 6alkyl, aryl sulfonyl C 1 6alkyl, aryl sulfonyl, aryl Cl 6 alkylsulfonyl, aryl C1 6 alkylsulfonyl Cl 6alkyl, wherein alkyl groups and aryl groups may be un.substituted or substituted with one or more substituents selected from R1 and R2;
W O 97/262~0 PCTrUS97100572 R6 is hydrogen, Cl ~ alkyl, or aryl, S aryl Cl 8 alkyl;
m is an integer selected from O to 6; and n is an integer selected from O to 6.
In a subgroup of the group are compounds having the formula 1~l 1~l 14 X~(CH2)m~Y~(CH2)n~C~IN-CH2-C-NH-CH-ICH-CO2R6 and pharmaceutically acceptable salt.s thereof, wherein X is S~-- ~/ ~/ H2N
H~l N ~ J ,~
N~N~ ~,~ or W 097/26250 PCT~US97/00572 - 2~ -Y is -(CH2)0 4-, 0 -N- , or -N- ;
R3 is hydrogen, S methyl, , or R4 is hydrogen, 1 0 methyl, 2 ~
~' ~
=CH;
W O 97/262~0 PCTAUS97/00572 , - 29 -.,R5 i,s hydrogen, or S R6 j~
hydrogen, methyl, ethyl, or t-butyl;
m i.s an integer se3ected from 0 to 6; and n i,s an integer selected from 0 to 6.
Specific examples of this subgroup include 1 5 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-alanine t-butyl ester 4-(2-~minothiazol-4-yl)butanoyl-glycyl-2(S~-phenylsulfonamido-~(3-alanine 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl)-,13-alanine methyl ester 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl~-,13-alanine 25 trifluoroacetate salt 5 -(2-Pyridylamino)pentanoylglycyl -2(S)-phenylsulfonamido-,13 -alanine ethyl ester 30 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine trifluoroacetate ~alt W 097/26250 PCTrUS97/OOS72 4-(2-Bocamino-pyridin-6-yl)butanoyl-~arcosine-3(R)-[(2-indol-3-yl)ethyl]-~3-alanine ethyl e,~ter 4-(2-Aminopyridin-6-yl)butanoyl-~sarcosine-3(R)-[(2-indol-3-yl)ethyl] -5 ,B-alanine 4-(2-Boc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsul~onamido-~-alanine t-butyl ester 10 4-(2-Aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,B-alanine 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-13-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-sarcosine-3(~ 2-(indol-3-yl)ethyl]-~-alanine 4-(2-Bocamino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3 (R)-(2-phenethyl)-~-alanine ethyl ester 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-~-alanine ethyl ester hydrochloride 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R~-(2-25 phenethyl)-,~-alanine 4-(2-Boc-amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol -3 -yl)ethyl] -,B-alanine 30 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-~(2-indol-3-yl)ethyl] -,13-alanine 4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-,(3-alanine ethyl ester W O 97126250 PCT~US97/00572 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-,B-alanine ethyl ester 4-(2-~mino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~3-S alanine 4-(Pyridin-4-yl)butanoyl-N-(2-phenylethyl)glycyl-3(R)-(2-phenethyl)-,13-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-13-alanine 4-(2-~3OC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-,B-alanine benzyl ester 4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-~-alanine 4-(2-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-20 ~-alanine 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanine ethyl ester 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-~3-alanir~e 3 -[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-,B-alanine ethyl ester 3-~(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-~3-alanine W O 97/26250 PCT~US97/00572 N- ~ N'-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl ~ -3(R)-rnethyl-,B-alanine benzyl ester N-[N'-~3-( 1 -Piperazinyl)l~enzoyl]glycyl]-3(R)-methyl-~-alanine 5 trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-,(3-alanine methyl ester 10 N-~N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-,B-alanine trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]-N'-(2-phenethyl)glycyl]-3~R)-(2-phenethyl)-13-alanine methyl ester 1~
N-[N'-[3-( 1 -Piperazinyl)benzoyl]-N -(2-phenethyl)glycyl] -3(R)-(2-phenethyl)-13-alanine trifluoroacetic acid salt 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanol-glycyl-~-a~anine 20 t-butyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)but~noyl-glycyl-~-alanine 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-pyridin-3-yl-~3-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)pyridin-3-yl-~-alanine Ethyl N-pyridin-4-ylisonipecotyl-N-cyclopropylglycine-3(S)-ethynyl-,~-alanine W O 97/26250 PCTrUS97/00572 N-Py~idin-ylisonipecotyl-N-cyclopropylglycine-3 (S)-ethynyl-~-alanine Ethyl N-pyridin-4-ylnipecotyl-N-cyclopropylglycine-3(S)-ethynyl-~-S alanine N-Pyridin-4-ylnipecotyl-N-cyclopropylglycine-3(S)-ethynyl-,~-alanine 10 4-(1,~',3,4-Tetrahydro- 1 ,~-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)gly-3(S)ethynyl-~B-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3 (S)-ethynyl-~-alanine, 3- ~ 2-~5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino]-acetylamino } -3(S)-pyridin-3-yl- propionic acid.
and pharmaceutically acceptable salts, such as the trifluoroacetate salt 20 and the hydrochloric acid salt Compounds of the invention are ak;o useful for inhibiting the bone resorption activity of mammalian osteoclast cells by a~1mini~tering a pharmacologically effective amount of the compound to a patient in need of such activity to inhibit the activity of mammalian 25 osteoclasts.
Compounds of the invention are also useful for inhibiting tumor growth in m~mmals. Pharrnacologically effective amounts of the compounds, including pharamaceutically acceptable salts thereof, are administered to the m~mmal, to inhibit tumor growth. The growth of 30 tumors depends on an adequate blood supply, which in turn depends on growl:h of new vessels into the tumor. New vessels are stimulated by factors ,secreted by the tumor. Inhibition of angiogenesis can cause tumor regres.sion in animals.
W O 97/26250 PCT~US97/00572 Compounds of the invention are also useful for treating and preventing diabetic retinopathy in m;~mm~ls. Pharmacologically effective amount,~ of the compounds, including pharamaceutically acceptable salts thereof, are ~lministered to the mammal, to inhibit diabetic re~inopathy.
Compounds are also useful in the prevention of restenosis of vessels.
The te~n "bone resorption activity" means the process by which osteoclast~ solubilize bone minerals and increase the activity of 10 en~ymes that degrade bone matrix.
Compounds of the invention are u,seful for inhibiting the binding of fibrinogen to blood platelets and for inhibiting the aggregation of blood platelets. The above-mentioned compounds can be used in a method of acting upon a fibrinogen receptor which comprises 15 administering a therapeutically effective but non-toxic amount of such compound to a m~mm~l, preferably a human. A pha~naceutical composition comprising a pharmaceutically acceptable carrier and, dispersed therein, an effective but non-toxic amount of such compound i~; another feature of this invention.
The invention also includes the u~;e of a compound of the invention, or a phalmaceutically acceptable .salt thereof, in the manufacture of a medicament for inhibiting the aggregation of blood platelets, preventing platelet thrombosis, preventing thromboembolism or preventing reocclu~ion, in a m~mm~l.
~ETAIL~D DESCRIPTION OF THE INVENTION
Fibrinogen receptor antagonist compounds of Formula I
are useful in a method of inhibiting the binding of fibrinogen to blood platelets and for inhibiting the aggregation of blood platelet~.
30 Fibrinogen receptor antagonists of this invention are illu~trated by compounds having the folmula: , The following compoullds were tested and found to inhibit platelet aggregation with IC50 values between about 0.01 ~M and 100 ,uM.
W 097/262S0 PCTrUS97/00572 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,13-alanine t-butyl ester 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-S alanine 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-~-alanine methyl ester 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-~2-phenethyl)-~-alanine trifluoroacetate ~alt 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenyl~ulfonamido-~-alanine ethyl ester 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine trifluoroacetate salt 4-(2-Bocamino-pyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-20 yl)ethyl]-~-alanine ethyl ester 4-(2-Aminopyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-yl)ethyl] -,B-alanine 4-(2-Boc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,~3-alanine t-butyl ester 4-(2-Aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-13-alanine 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-~2-(indol-3-yl)ethyl]-~-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-~3-alanine W O 97/262~0 PCT~US97/00572 4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-,B-alanine ethyl ester 4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3(R)-(2-~S phenethyl)-,B-~lanine ethy~ e~ter hydrochloride 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-13-alanine 1 0 4-(2-Boc-amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol -3-yl)ethyl] -~-alanine 4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3 (R)-~(2-indol-3 -yl)ethyl] -,~-alanine 4-(2-Bocamino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3(R)-methyl-~-alanine ethyl ester 4-(2-Amino-pyridin-6-yl)butanoyl -N-cyclopropylglycyl-3 (R)-methyl-,(3-20 alanine ethyl e~ter 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3 (R)-methyl-~-alanine 25 4-(Pyridin-4-yl)butanoyl-N-(2-phenylethyl)glycyl-3(R~-(2-phenethyl)-~-alanine ethyl ester 4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-~-alanine 4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3 (R)-methyl-~-alanine benzyl ester . CA 02242877 1998-07-13 W O 97/2625~ PCT~US97/OOS72 4-(2-lBOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-- methyl-~3-alanine 4-(2-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3 (R)-methyl-S ~-alanine 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanine ethyl ester 1 0 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,13-alanine 3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarco.sine-3(R)-(2-phenethyl)-~-alanine ethyl ester 3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-,(3-alanine N- { N '-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl } -3(R)-20 methyl-,13-alanine benzyl ester N-[N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-methyl-}3-alanine trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl ester N-[N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-~B-alanine trifluoroacetic acid salt N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]-N'-(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl ester - 3~ -N-[N'-L3-( 1 -Piperazinyl)benzoyl]-N -(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine trifluoroacetic acid salt 4-(1,2,3 ,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)butanol-glycyl-,(3-alanine 5 t-butyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)butanoyl-glycyl-,B-alanine 10 4-(1 ,2 ,3 ,4-Tetrahydro- 1 ,X -naphthyridin -7 -yl)butanoyl -glycyl -3 (S )- pyridin-~-yl-,~-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,g-naphthyridin-7-yl)butanoyl-glycyl-3(S)pyridin-3-yl-~(3-alnine Ethyl N-pyridin-4-yli~onipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-alanine N-Pyridin-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-20 alanine Ethyl N-pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~3-alanine 25 N-Pyridin-4-ylnipecotyl -N -cyclopropylglycyl-3 (S )-ethynyl -~13 -alanine 4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3(S)ethynyl-,B-alanine ethyl ester 4-(1 ,2,3,4-Tetrahydro- 1 ,~s-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3 (S)-ethynyl-,B-alanine .
W O 97/26250 PCTrUS97/00572 One test which is used to evaluate fibrinogen receptor antagonist activity i,s based on evaluation of inhibition of ADP-stimulated platelets. Aggregation requires that fibrinogen bind to and occupy the platelet fibrinogen receptor site. Inhibitors of fibrinogen 5 binding inhibit aggregation. In the ADP-stimulated platelet aggregation assa~ u,~ed to determine inhibition associated with the compounds claimed in the instant invention, human platelets are isolated from fresh blood, collected into acid citrate/dextrose by differential centrifugation follo~wed by gel filtration on Sepharose 2B in divalent ion-free Tyrode's 1() buffer (pH 7.4) containing 2% bovine serum albumin.
Platelet aggregation is measured at 37~C in a Chronolog aggregometer. The reaction mixture contains gel-filtered human platelets (2 x 10P~ per ml~, fibrinogen ~100 micrograms per ml (ug/ml)), Ca2t (1 mM), and the compound to be tested. The aggregation is 15 initiated by adding 10 mM ADP 1 minute after the other components are added. The reaction is then allowed to proceed for at least 2 minutes. The extent of inhibition of aggregation is expressed as the percentage of the rate of aggregation observed in the absence of inhibitor. The I~50 is the dose of a particular compound inhibiting 20 aggregation by 50% relative to a control lacking the compound.
The terrn "pharmaceutically acceptable salts" shall mean non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid. Representative salts include the following salts:
25 acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clav~lanate, citrate, dihydrochloride, edetate, edisylate, estolate, e,sylate, fumarate, gluceptate, gluconate, glllt~m~te, glycollylarsanilate, hexyLresorcinate, hydrabamine, hydrobromide, hydrochloride, 30 hydroxynapthoate, iodide, isothionate, lactate7 lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote, palmitate, panthothenate, phosphate/diphosphate, polygalacturonate, W O 97/26250 PCTrUS97100572 salicylate, stearate, ,~ubacetate, ~uccinate, tannate, tartrate, teoclate, tosylate, triethiodide, trifluoroacetate, ;md valerate.
Compounds of the present invention are chiral; included within the ,~cope of the pre~ent invention are racemic mixtures and separated enantiomers of the general formula. Furthermore, all dia~tereomers, including E, Z isomers, of the general formula are included in the pre~ent scope. Furthermore, hydrates as well as anhydrous compositions and polymorphs of the general formula are within the present inventioIl.
Prodrugs, ,~;uch a,~; ester derivatives of described compounds, are compound derivatives which, when absorbed into the bloodstream of a warrn-blooded ~nim;~l, cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.
lS The term "pharmaceutically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the biological or medical respon,se of a tissue, system or animal that is being ,sought by a researcher or clinician. The term "anti-coagulant" shall include heparin, and warfarin. The term "thrombolytic agent" shall 20 include agents such as streptokina,se and tissue pla,sminogen activator.
The term "platelet anti-aggregation agent" ~hall include agents such as aspirin and dipyridamole.
The term "alkyl" means straight or branched alkane containing 1 to about 10 carbon atoms, e.g., methyl, ethyl, n-propyl, 25 isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexy, octyl radicals and the like, straight or branched alkene containing 2 to about 10 carbon atoms, e.g., propylenyl, buten-l-yl, isobutenyl, pentenylen- I -yl, 2~2-methylbuten- 1 -yl, 3-methylbuten- 1 -yl, hexen- I -yl, hepten-l-yl, and octen-1-yl radicals and the like, or straight or branched 30 alkyne cont~ining 2 to about 10 carbon atoms, e.g.~ ethynyl, propynyl, butyn-l-yl, butyn-2-yl, pentyn-l-yl, pentyn-2-yl, 3-methylbutyn-1-yl, hexyn-l-yl, hexyn-2-yl, hexyn-3-yl, 3,3-dimethylbutyn-1-yl radical,~; and the like.
The term "aryl" means a 5- or 6-membered aromatic ring c conta-;ning 0, 1, or 2 heteroatoms selected from O, N, and S. Examples of aryl include phenyl, pyridine, pyrimidine, imidazole, thiophene, oxazole, isoxazole, thiazole, and amino- and halogen- substituted derivatives thereof.
The te~n~ "alkyloxy" or "alkoxy" include an alkyl portion where alkyl is as defined above. Examples of alkoxy include methyloxy, propyloxy, and butyloxy.
The term.s "arylalkyl" and "alkylaryl" include an alkyl portion where alkyl is as defined above and to include an aryl portion whe~ aryl i~ as defined above. The Co-n or (~l-n designation where n may be an integer from 1-10 or 2-10 respectively refer~ to the alkyl component of the arylalkyl or alkylaryl unit. Examples of arylalkyl include benzyl, fluorobenzyl, chlorobenzyl, phenylethyl, phenylpropyl, fluorophenylethyl, chlorophenylethyl, thienylmethyl, thienylethyl, and thienylpropyl. Examples of alkylaryl include toluene, ethylbenzene, propylbenzene, methylpyridine, ethylpyridine, propylpyridine, hutylpyridine, butenylpy~idine, and pentenylpyridine.
The term "halogen" includes fluorine, chlorine, iodine and bromine.
The term "oxy" means an oxygen (O) atom. The term "thio" means a sulfur (S) atom. Under standard nonmenclature used throughout this disclosure, the terminal portion of the designated side chain is described first followed by the adjacent functionality toward the point of attachment. For example, a C1 6 alkyl substituted with C1 5 alkyl-carbonylamino i.s e4uivalent to HO
C1 6-alkyl-N-C-C1 s-alkyl Compounds of the invention where X i~s a 5-membered monocyclic aromatic ring system, e.g., a thiazole system, can be prepared by forming an alkyl ester stituted derivative of the ring, 35 e.g., methyl 4-(2-aminothiazol-4-yl)butanoate, forming the corresponding acid with HCl, and reacting with an amine to form the final product.
Compounds of the invention where X is a 6-membered monocyclic aromatic ring system, e.g., a pyridine Isystem, can be 5 prepared using 2-aminopyridine, 2-aminopicoline, 4-vinyl pyridine, etc., as described in Schemes 3, 4, and 10.
Compounds of the invention where X is a 9-membered polycyclic aromatic fused ring system can be prepared by reacting a substituted 5-membered ring starting material such as 2-amino-3-bromo 10 thiophene, 2-nitro-3-bromo thiophene, 2-amino-3-bromo pyrrole, and 2-amino-3-bromo furan, with an appropriate compound under suitable ring clo,sure conditions to effect formation of the 9-membered fused ring system.
Compounds of the invention where X is a 10-membered 15 polycyclic aromatic ring sy,stem can be prepared using a starting material such a~s naphthyridin (Hamada, Y. et al., Chem. Pha7 m. Bull.
Soc., 1971, 19(9), 1857-1862), or by reacting an aminoaldehyde pyridine with a .suitable ketone under suitable ring closure conditions to effect formation of the 10-membered fused ring system.
The examples illustrate procedures for preparing compounds of the invention where Y is -(CH2)0 4, -O-~ and -N(R~)-.
To make compounds where Y is -N(R~)C(O)-, an acid such as compound 1-4 can be subjected to a Curtius reaction to form the amine, and subsequent condensation to give the final product.
In the schemes and examples below, various reagent symbols have the following meanings:
BOC
(or Boc): t-butyloxycarbonyl Pd-C: Palladium on activated carbon catalyst DMF: Dimethylformamide DMSO: Dimethyl,sulfoxide CBZ: Carbobenzyloxy CH2Cl2: Methylene chloride W 097/2625(~ PCT~US97/00572 CHC13: chloroform r EtOH: ethanol MeOH: methanol EtOAc: ethyl acetate S HOAc: acetic acid BOP. Benzotriazol- l -yloxytris(dimethylamino)phosphonium, hexafluorophosphate EDC: 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride Oxone: potassium peroxymonosulfate LDA: Lithium diisopropylamide PYC]LU: Chloro-N,N,N',N'-bis(pentamethylene)formamidinium hexafluorophosphate l~ The compounds of the present invention can be ~1mimistered in such oral forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granule~., elixirs, tinctures, suspensions, syrups, and emulsions.
Like~ise, they may be ~(lmini~tered in intravenous (bolus or infusion), intra~)eritoneal, subcutaneous, or intramusculsar form, all using forms well Iknown to those of ordinary skill in the pharmaceutical arts. An effec~ive but non-toxic amount of the compound desired can be employed as an anti-aggregation agent.
Compounds of the invention may be administered to patients where prevention of thrombosis by inhibiting binding of fibrinogen to the platelet membrane glycoprotein complex IIb/IIIa receptor is desired. They are useful in surgery on peripheral arteries (arteIial grafts, carotid endarterectomy) and in cardiovascular surgery where manipulation of arteries and organs, and/or the interaction of r 3Q plate]ets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelets may form thrombi and thromboemboli. Compounds of this invention may be ~lmini~tered to these surgical patients to prevent the formation of thrombi and thromboemboli.
W O 97/26250 PCT~US97100572 Extracorporeal circulation is routinely used for cardiovascular surgery in order to o~ygenate blood. Platelets adhere to surfaces of the extracorporeal circuit. Adhesion is dependent on the interaction between gp rIb/IIIa on the platelet membrane,~ and 5 fibrinogen adsorbed to the surface of the circuit. (Gluszko et al., Ame7 .
~1. Physiol., 252(H), 615-621 (1987)). Platelets released from artificial surfaces show impaired hemostatic function. Compounds of the invention may be admini,stered to prevent adhesion.
Other applications of these compounds include prevention 10 of platelet thrombosis, thromboembolism and reocclusion during and after thrombolytic therapy and prevention of platelet thrombosis, thromboembolism and reocclusion after angioplasty or coronary artery bypass procedures. They may also be used to prevent myocardial infarction.
The dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of a-lministration; the renal and hepatic function of the patient; and the particular compound 20 or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug re~luired to prevent, counter, or arrest the progress of the condition.
Oral dosages of the present invention, when used for the 2~ indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 100 mg/kg/day and preferably 0.01-SQ mg/kg/day and more preferably 0.01-20 mg/kg/day, e.g. 0.1 mg/kg/day, 1.0 mg/kg/day, 5.0 mg/kg/day, or 10 mg/lcg/day.
Intravenously, the most preferred doses will range from about 1 to 30 about 10 mg/kg/minute during a constant rate infusion.
Advantageously, compounds of the present invention may be administered in divided doses of two, three, or four times daily.
Furthermore, preferred compounds for the present invention can be ~ministered in intranasal form via topical use of suitable intranasal W O 97/26250 PCTrUS97/00572 vehicles, or via transdermal routes, using those forms of transderrnal skin patches well known to those of ordinary skill in that art. To be a~lmini~stered in the form of a transdermal delivery system, the dosage ~lministration will, or course, be continuous rather that intermittent 5 throughout the dosage regime.
In the methods of the present invention, the compounds hereiIl described in detail can form the active ingredient, and are typically a(lmini~tered in admixture with suitable pharrnaceutical diluents, e~cipients or carriers (collectively referred to herein as 10 "carrier" materials) suitably selected with respect to the intended form of ~lministration, that is, oral tablets, capsules, elixirs, syrups and the like, cmd consistent with convention pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, 15 non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert 20 carrier such as ethanol, glycerol, water and the like. ~oreover, when desired or necessary, suitable binder,s, lubricants, distintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lacto~e, corn-sweeteners, natural and synthetic gums such as acacia, 25 tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms inclucle sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators inclucle, without limitation, starch methyl cellulose, agar, bentonite, 30 xanthan gum and the like.
., The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar ve,sicles, large unilamellar vesicles and multilamellar W 097126250 PCTrUS97/00572 vesicles. Liposomes can be forrned from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
Compounds of the present invention may also be delivered by the u,se of monoclonal antibodies as individual carriers to which the ;S compound molecule~ are coupled. The compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copo}ymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxy-ethyl-aspartamide-phenol, or polyethyleneoxide-polylysine sub,stituted 10 with palmitoyl residue~s. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled relea,se of a drug, for example, polylactic acid~
polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, 1~ polyacetals, polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block copolymers of hydrogels.
The compounds of the present invention can also be co-administered with suitable anticoagulation agents or thrombolytic agents such a~ plasminogen activators or streptokinase to achieve synergistic 20 effect.~ in the treatment of various vascular pathologie.s. They may also be combined with heparin, aspirin, or warfarin.
The novel compounds of the pre.sent invention were prepared according to the procedure of the following examples. The most preferred compounds of the invention are any or all of those 25 specifically set forth in these examples. These compounds are not, however, to be construed as forming the only genus that is considered as the invention, and any combination of the compounds or their moieties may itself form a genus. The following examples further illustrate detail,s for the preparation of the compounds of the present invention.
3(~ Tho.se skilled in the art will readily understand that known variations of the condition.s and processes of the following preparative procedures can be used to prepare these compounds. All temperatures are degrees Cel.siu.s unless otherwise noted.
W O 97/26250 PCT~US97/00572 SCHEME I
~ ~i CO2H HCI-H2N~H SO
Br2, MeOH
0~C to RT Boc-Gly-OH
r NMM, isobutyl chloroformate, Br~~/~CO2Me O EtOAc, -t5~Cto RT
0 1-2 BocHN~J~ N~CO2-t-Bu ,, EtOH, ~ H H NH--S~2 H~N~<N~ CO2Me TFA, CH2ci2 1-3 -1 5~C
r TFA H2N~ll~N ~C02-t-Bu H H ' NH--S~2 HCi~ i l2N y,N~ c02H 1-7 EDC, HOBT, NMM
DM F, -1 5~C to RT
WO 97/26250 PCTrUS97/00572 - 4~ -H2N y/ ~ N ~J~ N ~CO~-t-Bu~
1 -~
TFA, CH2CI2 H2Ny/~ ~H 2 Br~~CO2CH3 ~ 1-2 Methyl 6-bromo-5-oxohexanoate (1-2) 5-Oxohexanoic acid (1-1, 5 mL, 42 mmol) was dissolved in ~S4 mL MeOH and cooled to 0~C. Br2 (2.2 mL, 43 mmol) was added 10 dropwise, and the reaction was stirred at RT overnight. After removing the MeOH by rotary evaporation, the residue was dissolved in ether, washed with water, sat. NaHCO3 and brine, dried (Na2SO4), filtered and concentrated. Flash chromatography (silica, 10% EtOAc/hexane) provided the bromide-ester 1 2 as a yellow oil.
1~ TLC Rf 0.09 (silica, 15% EtOAc/hexane) lH-NMR (300 MHz, CDC13): ~ 3.8~s (s, 2H), 3.67 (s, 3H), 2.75 (t, J =
7Hz, 2H), 2.37 (t, J = 7 Hz, 2H), 1.94 (qn, J = 7 Hz, 2H).
W 0971262~ PCT~US97100572 y ~ co2CH3 Meth~l 4-(2-aminothiazol-4-yl)butanoate ( 1-3) Bromide 1-2 (3.45 g, 15.5 mmol) and thiourea (1.4 g, 18 mmol) were combined in 77 mL EtOH and heated to reflux. After disappearance of 1 2, the EtOH was removed by rot~ry evaporation and the residue was diluted with EtOAc, washed with water and brine, then dried (MgSO4), filtered and concentrated. The pH of the aqueous phase wa~ adjusted to 7, and the solution was re-extracted with EtOAc (2x).
These organic extract~s were washed with brine, dried (MgSO4), filteled and concentrated, combined with the first organic residues, then purified by flash chromatography (silica, EtOAc) providing aminothiazole 1 3 as a white solid.
Tl,C Rf 0.5 (silica, EtOAc) lH-NMR (400 MHz, CDCl3): ~ 6.09 (s, lH), 5.19 (br s, 2H), 3.66 (s, 3H~, 2.55 (t, J = 7 Hz, 2H), 2.34 (t, J = 7 Hz, 2H), 1.96 (qn, J = 7 Hz, 2H).
HCi H2N Y'~' --CO2H
4-(2-,~minothiazol-4-yl)butanoic acid hydrochloride (1-4~
Ester 1 3 (1.3 g, 6.5 mmol) was dissolved in 32 mL 6 N
H~l. After stirring overnight, the resulting suspension was concentrated, providing acid 1 4 as a white solid.
1H-NMR (400 MHz, d6-DMSO): o 9.12 (br s, lH), 6.51 (s, lH), 3.50 (br s), 2.51 (t, J = 7 Hz, 2H), 2.24 (t, J = 7 Hz, 2H), 1.77 (cln, J = 7 Hz, 2H).
W O 97126250 PCTrUS97/00572 o BOCNH NH ~CO2-t-BU
H NHSO2Ph N-Boc-glycyl-2(S)-phenyllsul~onamido-,B-alanine t-butyl ester (1-6) N-Boc-glycine (255 m~, 1.5 mmol) wa~ dissolved in 7.4 mL EtOAc, cooled to -15~C, then NMM (179 ~L, 1.6 mmol) and S isobutyl chloroformate (211 ~lL, 1.6 mmol) were added. After 20 min, amine 1 5 (500 mgt 1.5 mmol) and additional NMM (422 ,uLt 3.2 mmol) were added and the reaction was wa~ed to RT overnight.
l~ollowing dilution with EtOAc, the mixture was washed with water, sat.
NaHCO3, 10% KHSO4, and brine, dried (MgSO4), filtered and concentrated, providing amide 1 6 as a white solid.
TLC Rf 0.73 (silica, EtOAc) lH-NMR (300 MHz, CDC13): o 7.g4 (d, J = 7 Hz, 2H), 7.59 (ABX t, J =
DM F, -1 5~C to RT
WO 97/26250 PCTrUS97/00572 - 4~ -H2N y/ ~ N ~J~ N ~CO~-t-Bu~
1 -~
TFA, CH2CI2 H2Ny/~ ~H 2 Br~~CO2CH3 ~ 1-2 Methyl 6-bromo-5-oxohexanoate (1-2) 5-Oxohexanoic acid (1-1, 5 mL, 42 mmol) was dissolved in ~S4 mL MeOH and cooled to 0~C. Br2 (2.2 mL, 43 mmol) was added 10 dropwise, and the reaction was stirred at RT overnight. After removing the MeOH by rotary evaporation, the residue was dissolved in ether, washed with water, sat. NaHCO3 and brine, dried (Na2SO4), filtered and concentrated. Flash chromatography (silica, 10% EtOAc/hexane) provided the bromide-ester 1 2 as a yellow oil.
1~ TLC Rf 0.09 (silica, 15% EtOAc/hexane) lH-NMR (300 MHz, CDC13): ~ 3.8~s (s, 2H), 3.67 (s, 3H), 2.75 (t, J =
7Hz, 2H), 2.37 (t, J = 7 Hz, 2H), 1.94 (qn, J = 7 Hz, 2H).
W 0971262~ PCT~US97100572 y ~ co2CH3 Meth~l 4-(2-aminothiazol-4-yl)butanoate ( 1-3) Bromide 1-2 (3.45 g, 15.5 mmol) and thiourea (1.4 g, 18 mmol) were combined in 77 mL EtOH and heated to reflux. After disappearance of 1 2, the EtOH was removed by rot~ry evaporation and the residue was diluted with EtOAc, washed with water and brine, then dried (MgSO4), filtered and concentrated. The pH of the aqueous phase wa~ adjusted to 7, and the solution was re-extracted with EtOAc (2x).
These organic extract~s were washed with brine, dried (MgSO4), filteled and concentrated, combined with the first organic residues, then purified by flash chromatography (silica, EtOAc) providing aminothiazole 1 3 as a white solid.
Tl,C Rf 0.5 (silica, EtOAc) lH-NMR (400 MHz, CDCl3): ~ 6.09 (s, lH), 5.19 (br s, 2H), 3.66 (s, 3H~, 2.55 (t, J = 7 Hz, 2H), 2.34 (t, J = 7 Hz, 2H), 1.96 (qn, J = 7 Hz, 2H).
HCi H2N Y'~' --CO2H
4-(2-,~minothiazol-4-yl)butanoic acid hydrochloride (1-4~
Ester 1 3 (1.3 g, 6.5 mmol) was dissolved in 32 mL 6 N
H~l. After stirring overnight, the resulting suspension was concentrated, providing acid 1 4 as a white solid.
1H-NMR (400 MHz, d6-DMSO): o 9.12 (br s, lH), 6.51 (s, lH), 3.50 (br s), 2.51 (t, J = 7 Hz, 2H), 2.24 (t, J = 7 Hz, 2H), 1.77 (cln, J = 7 Hz, 2H).
W O 97126250 PCTrUS97/00572 o BOCNH NH ~CO2-t-BU
H NHSO2Ph N-Boc-glycyl-2(S)-phenyllsul~onamido-,B-alanine t-butyl ester (1-6) N-Boc-glycine (255 m~, 1.5 mmol) wa~ dissolved in 7.4 mL EtOAc, cooled to -15~C, then NMM (179 ~L, 1.6 mmol) and S isobutyl chloroformate (211 ~lL, 1.6 mmol) were added. After 20 min, amine 1 5 (500 mgt 1.5 mmol) and additional NMM (422 ,uLt 3.2 mmol) were added and the reaction was wa~ed to RT overnight.
l~ollowing dilution with EtOAc, the mixture was washed with water, sat.
NaHCO3, 10% KHSO4, and brine, dried (MgSO4), filtered and concentrated, providing amide 1 6 as a white solid.
TLC Rf 0.73 (silica, EtOAc) lH-NMR (300 MHz, CDC13): o 7.g4 (d, J = 7 Hz, 2H), 7.59 (ABX t, J =
7 Hz, IH), 7.51 (ABX t, J = 7 Hz, 2H), 6.58 (br m, lH), 5.5~ (d, J = 8 Hz, lH), 5.11 (br s, lH), 3.90-3.7fs (m, 3H), 3.72 (m, llI), 3.40 (m, lH), 1.48 (.~, 9H), 1.2~ (s, 9H).
TFA ~H2N ~,1 1-7 NHSO2Ph Glycyl-2(S)-phenyl~ulfonamido-,B-alanine t-butyl ester trifluoroacetate 20 salt (1-7) Protected amide 1-6 (576 mg, 1.26 mmol) was dissolved in 6.3 mL CH2C12, cooled to -15~C, and TFA (6.3 mL) was added. After 25 min the reaction was concentrated, providin~ amine 1 7.
TLC Rf 0.36 (silica, 9:1:1 CH2C12/MeOH/HOAc).
W O 97/2625~ - - PCTr~S97/00572 H2N~<,N~NJ~ N ~CO2-t-Bu~
4-(2-Aminothiazol -4-yl)butanoyl-glycyl-2(S)-phenyl.sulfonamido-,B-alanine t-butyl ester (1 -g~
Acid ~-4 (300 mg, 1.35 mmol), amine 1 7 (600 mg, 1.37 5 mmol), HOBT (219 mg, 1.14 mmol) and NMM (445 ,uL, 4.04 mmol) were combined in 13 mL DMF, cooled to -15~C, and EDC (310 mg, 1.61 mmol) was added. The reaction was warmed to RT, stirred ovemight, then diluted with LtOAc, washed with water, sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash 1~ chromatography (silica, 20% MeOH/EtOAc) provided 1 ~ as yellow solid.
TLC Rf 0.55 (silica, 20% MeOH/EtOAc) 1H-r~MR (400 MHz, CDC13): ~ 7.80 (d, J = 7 Hz, 2H), 7.55 (ABX t, J =
7 Hz, lH), 7.47 (ABX t, J = 8 Hz, 2H), 7.35 (br s, lH), 7.04 (br m, lH), 6.12 (s, lH), 5.41 (br s, 2H), 4.05-3.95 (m, 3H), 3.69 (m, IH), 3.39 (ddd, lH), 2.70-2.55 (m, 2H), 2.33 (m, 2H), 2.01 (qn, J = 7 Hz, 2E~), 1.27 (~;, 9~) H2N y~N~ N N ~~2H
S C) H H ~ S0 4-(2-Aminothiazol-4-yl)butanoyl-glycyl -2(S)-phenylsulfonamido-,B-alaninç (1 -9) ~- Ester 1 ~s ~365 mg, 0.69 mmol) was dissolved in CH2cl2 (3.5 mL), then TFA (3.5 mL) was added. After 5 h the reaction 25 mixture was concentrated, azeotroped with toluene, then purified by sequential flash chromatography (silica, 22:20: 1: 1 W O 97/26250 PCT~US97/00572 EtOAc/EtOH/H20/NH40H, then silica, 4:1:1 CH2(~12/MeOH/HOAc, then 7:1:1 CH2cl2lMeoHlHoAc)~ providing 1 9 as a white solid.
TLC Rf 0.33 (silica, 7:1:1 CHzC12/MeOH/HOAc) 1 H-NMR (400 MHz, CD30D): ~ 7.~s6 (d, J - 7 Hz, 2H), 7.5~ (ABX t, J
= 7 Hz, lH), 7.52 (ABX t, J = 8 Hz, 2H), 6.27 (s, lH), 3.~,9 (AB d, J =
17 Hz, lH), 3.77 (AB d, J = 17 Hz, lH), 3.64 (t, J = 6 Hz, lH), 3.53 (AB dd, lH), 3.41 (AB dd, lH), 2.57 (t, J = 7 Hz, 2H), 2.35-2.25 ~m, 2H~, 1.95 (m, 2H).
L-asparagine NaOH, H20, PhSO2CI
H2NC--~
H NHSO2Ph NaOH, dioxane, Br2 0~C to 90~C
-.. H2 \1 ~,CO2H
H N HSO2Ph - 1-2a a. isobutylene, H2SO4, dioxane / SOCI2 \b. 1N HCI/ether / ethanol HC~ ~ H2N ~6 2 ~~
H NHSO2Ph H NHSO2Ph N-Phenylsulfony~-L-a~paragine ( 1 - l a) To a stirred solution of L-asparagine (Aldrich) (lO g, 76 mmol), NaOH (3.4 g, 85 mmol), H20 (50 mL), and dioxane (50 mL) at 0~C was added PhS02CI (10.6 mL, ~4 mrnol). After 1 min, NaOH (3.4 g) in ~2~ (50 mL) was added and the reaction mixfure stirred for 30 min. The reaction mixture was then concentrated to remove the dioxame then washed with EtOAc. ~he aqueous phase was then cooled to 0~C and acidified to pH 5.0 with conc. HCI to effect product precipitation. The re,sulting solid was collected by filtration, washed with II20 (20 mL) and dried at 50~C under vacuum to give N-phenylsulfonyl-L-asparagine (I-la) as a white solid.
~f 0.40 (silica, l O: 1:1 ethanol/H20/NH40H). I H NMR (300 MHz, D20) ~ 7.59 (m, 2H), 7.26 (m, 3H), 3.92 (m, lH), 3.02 (m, lH), 2.35 (m, l]H).
W 097/26250 PCT~US97100572 H2N ~6 C~2H
H N HSO2P h 3-Amino-2(S)-phenylsulfon~laminopropionic acid (1-2b) To stirred solution of NaOH (15.6 g, 0.4 mol) in H2O (70 mL)1 cooled with an icebath, was added bromine (3.6 mL, 0.07 mol) dropwise. After 5 min, a cold .solution of N-phenylsulfonyl-L-asparagine, l-la (14.6 g,54 mmol) and NaOH (4.3 g, 0.1 mol) in H2O
(50 mL) was added in one portion. The solution was stirred for 20 min at 0~C then 30 min at 90~C. The reaction mixture was recooled to 0~C, and the pH adjusted to 7 through dropwise addition of conc. HCl. The 10 white precipitate forrned was collected by filtration and then dried to give ~) as a white solid. lEI NMR (300 MHz, D2O) ~ 8.00, 7.50 (m, 5H), 3.8~s (m, lH), 3.37 (m, lH), 3.12 (m, lH).
2 ~6 H NHSOZPh 1~ Ethyl 3-Amino-2(S)-phenylsulfonylaminopropionate hydrochloride (3-4) Arnino acid 1 -2a ( 1.0 g, 4.1 mmol) was suspended in 20 mL EtOH, cooled to 0~C, and SOC12 (1.5 mL, 21 rnrnol) was added dropwise. After stirring at RT overnight the mixture was concentrated, 20 triturated with Et2O (2x), and dried, providing 3-4 (1.26 g) as a hygroscopic yellow solid.
lH-NMR (300 MHz, d6-DMSO): ~ ~.30 (br s), 7.79 (d, J = ~ Hz, 2H), 7.70-7.60 (m, 3H), 4.21 (t, J Hz, lH), 3.90-3.~0 (m, 2H), 3.09 ~ABX
dd, J = 13, 6 Hz, lH), 2.90 (ABX dd, J = 13, ~ Hz, 2H), 0.97 (t, J = 7 2~ Hz, 3H).
W O 971262S0 PCT~US97/00~72 HCI- H2N~
H NHSO2Ph tert-Butyl 3-Amino-2(S)-phenylsulfonylaminopropionate hydro-chloride f 1-~) In a Fischer-Porter tube, a mixture of 1-2a (10.2 g, 42 mmol) and DME~ (150 mL) was sequentially treated with ~2S04 (6.4 mL, ~.12 mol), cooled to -7~~C, and then condensed isobutylene (75 mL). The cooling bath was removed. After 24 h, ice/water (250 mL) was added ~ollowed by washing with ether (2x). The a~ueous phase was basified with aq 6N NaOH, then saturated with NaCI, followed by extraction with EtOAc (3x). The combined extracts were washed with brine, dried (MgS04), and concentrated to give a white .solid. This was dissolved in CH2C12 and treated with lN HCl/ether (22 mL), and then concentrated to give 1 5 as a glassy yellow solid.
lH NMR (400 MHz, DM~O) o 8.25-8.00 (m, 4H), 7.85-7.5fs (m, 5H), 4.0~ (m, lH), 3.10 (m, IH), 2.73 (m, lH), 1.17 (s, 9H).
W O 97126250 PCT~U~97/00572 ~, HCI-H2Ny~ ~CO2H O J
1-4 HCI-H2N ~J~ ~ CO2Me ~ / 2-1 EDC, HOBT, NMM
DMF, -1 5~C to RT ~
y ~ N ~J~ N ~CO2Me 1 NNaOH
MeOH ¢~
TFA H2Ny~N~ ~ H H ~
S O
~ A
W O 97/2625~ PCTMS97/00572 . - 57 -~ H (CH2)2Ph NHJlNH~ C02CH3 S O
4-(2-Aminothiazol -4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl)-~3-alanine methyl ester (2-2) Acid 1 4 (300 mg~ 1.35 mmol), amine 2-1 (405 mg, 1.35 mmol) (prepared as described in Duggan et al., U.s. Patent 5,264,~20) HOBT (219 mg, 1.62 mmol) and NMM (445 ~L, 4.04 mmol) were combined in 7 mL DMF, cooled to -15~C, and EDC (310 mg, 1.61 mmol) was added. The reaction was warmed to RT, stirred overnight, 10 then cliluted with EtOAc, washed with water, ~sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography ~ilic.l, 10% MeOH/EtOAc) provided 2-2 as a yellow oil.
TLC Rf 0.32 (silica, 10 % MeOH/EtOAc) 1 H-NMR (400 MHz, CDC13): o 7.82 (d, J = 7 Hz, 1 H), 7.65 (d, ~ = ~
1~ Hz, lH), 7.40-7.10 (m, 5H), 6.93 (d, J = 8 Hz, lH), 6.10 (s, lH), 4.31 ~m, IH), 3.96 (ABX dd, J = 17, 6 Hz, lH), 3.89 (ABX dd, J = 17, 5 Hz, lH), 3.64 (s, 3H), 2.68-2.54 (m), 2.32-2.17 (m, 2H), 2.25~ 0 (m).
~ H (CH2)2Ph ~NHJlNH~CO2H
S O
~ 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-~-alanine trifluoroacetate salt ~2-3) Ester 2-2 (220 mg, 0.51 mmol) and 1 N NaOH (1.3 mL, 1.3 mmol) were combined in 5 mL MeOH. After 3 d the reaction 25 mixture was concentrated, purified by flash chromatography (.~iilica, 9: 1: 1 CH2C12/MeOH/HOAc), then preparative HPLC (Cl g, 0.1 % TFA
W O 97/26250 PCTAUS97/00~72 - 5~ -in CH3CN/H20), providing, after Iyophilization, acid 2-3 a~ a white ~olid.
TLC Rf 0.54 (silica, 4:1:1 CH2C12/MeOH/HOAc) lH-NMR (400 MHz, CD30D): ~ 7.26-7.10 ~m, 5H), 6.52 (~, lH), 4.23 5 (m, lH), 3.~¢ (AB d, J = 17 Hz, IH), 3.79 (AB d, J = 17 Hz, lH), 2.72-2.5~ (m, 4H), 2.51 (d, J = 7 Hz, 2H), 2.34 (t, J = 7 Hz, 2H), 1.99-1.75 (m, 4H).
W O 97/262~,0 PCTAUS97/00572 ~, NH~ ~N SO~
Cbz-G Iy-OH
NaH, DMF, 0~C to RT; isobutyl chlorofor,nate, Ethyl 5-bromopentanoate NMM
~ RT to 75~C EtOAc, -15~C to RT
¢~N CO2E~ ~H
1 N NaOH, EtOH H2, 10 % Pd/C
EtOH
~, N~ N ~ CO2H ~
~J H2N J~ N ~CO2Et 3 3 \/ H H NH-SO
BOP, NMM
DMF
r W O 97/26250 PCTrUS97/00572 - 60 - v H H (~
N~ N ~ N~ N ~CO2Et oH H N - S0 LiOH, THF
TFA~ N'J~N ~C02H
NI~ ~ co2Et Ethyl 5-~2-pyridylamino)pentanoate (3-2) 2-Aminopyridine (3-1, 1.97 ~, 20.9 mmol) in 10 mL DMF
was added to a suspension of NaH (60 % in oil, 1.00 g, 25 mmol) in 80 mL DMF cooled to 0~C. After warming to RT for 45 min, ethyl 5-10 bromopentanoate (4.2 mL, 25 rnmol) wa~; added dropwi,se. Thismixture was heated at 75~C overnight, then cooled to RT, diluted with EtOAc, washed with water (2x), sat. NaHCO3, and ~rine, drie~
~MgSO4), filtered and concentrated. Flash chromatography (silica, 50%
then 70 % EtOAc/hexane) provided 3-2 as a yellow oil.
15 TLC R~ 0.55 (silica, 70 % EtOAc/hexane) H-NMR (400 MHz, CDC13): ~ ~.07 (dd, J = 5, I Hz, lH), 7.40 (m, lH), 6.55 (m, lH), 6.37 (d, J = ~ Hz, lH), 4.4~ (br s, lH), 4.13 (4, J = 7 Hz, 2H), 3.29 (q, J = 7 Hz, 2H), 2.35 (t~ J = 7 Hz, 2H), l.g0-1.55 (m 4H), 1.25 (t, J = 7 Hz, 3H).
W O 97126250 PCT~US97100572 ~,NH'--~--C02H
5-(2-Pyridylamino)pentanoic acid (3-3) Ester 3-2 (0.41 g, 1.~4 mmol) was dissolved in 18 mL
5 EtOH, 1 N NaOH (4.6 mL, 4.6 mmol) was added, and the reaction was stirrecl overnight. The pH of the solution was adjusted to 7 with I N
HCl, and concentration provided a white solid containing acid 3-3 and NaCI.
TLC Rf 0.06 (silica, l 9:1:1 CH2Cl2/MeOH/HOAc) lH-NMR (400 MHz, D2O): ~ 7.~1 (m, lH), 7.77 (d, J = 6 Hz, lH), 6.96 (d, J -- 9 Hz, lH), 6.~2 (t, J - 7 Hz, lH), 3.36 (t, J = 7 Hz, 2H), 2.24 (m, 2H), ~.72-1.50(m, 4H).
CbZNH~ NH~6co2Et !L5 N-Cb%-~lycyl-2(S~-phenylsulfonamido-,B-alanine ethvl ester (3-5) N-Cbz-glycine (339 mg, 1.62 mrnol) was dissolved in 8 mL
EtOAc, cooled to -15~C, then NMM (196 }IL, 1.~ mmol) and isobutyl chloroformate (230 ~lL, 1.8 mmol) were added. After 20 min, the 20 mixed anhydride solution was added to amine 3-4 (0.50 mg, 1.6 mmol) suspended in 5 mL EtOAc and the reaction was warrned to RT for 90 min. Following dilution with EtOAc, the mixture was washed with water, sat. NaHCO3, 5% KHSO4, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 75 % EtOAc/hexane) 25 provided amide 3-5 as a colorless oil.
TL~ ]R~ 0.29 (silica, 75 % EtOAc/hexane) W O 97/26250 PCTrUS97/00572 lH-NMR (300 MHz, CDC13): ~ 7.65-7.45 (m, 3H~, 7.40-7.25 (m, 5H), 6.6~ (t, J = 6 Hz, lH), 5.~s3 (d, J = ~ Hz, lH), 5.49 (t, J = 6 Hz, lH), 5.15 (.s, 2H), 4.04-3.95 (m, 3H), 3.~9-3.~5 (m, 2H), 3.73 (m, IH), 3.46 (m, lH), 1.11 (t, J = 7 Hz, 3H).
s O
H2N~NH ~CO2Et H- NHSO2Ph Glycyl-2(S)-phenyl.sulfonamido-~-alanine ethyl ester (3-6) Protected amine 3-5 (0.47 g, 1.01 mmol) was dissolved in 10 mL EtOH, 10 % Pd/C (94 mg) was added, and the reaction was stirred under an H2 balloon. After 4 h, additional 10 % Pd/C was added (94 mg), and the reaction was continued for 3 d. The mixture was filtered through Celite, concentrated, and azeotroped with CHC13, providing amine 3-6 as a gum.
l~-NMR (300 MHz, CDC13): ~ 7.95 (m), 7.~6 (d, J = 7 Hz, 2H), 7.60-7.45 (m, 3H), 4.05 (dd, J = 5, 6 Hz lH), 3.96 (q, J = 7 Hz, 2~), 3.g0-3.55 (m), 1.07 (t, J = 7 Hz, 3H).
[~,N H ~ ~6N HSO2Ph ~3-7 5 -(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine ethyl ester (3-7) Acid 3-3 (1~6 mg, 0.55 mmol), amine 3-6 (150 mg, 0.46 mmol), NMM (0.20 mL, 1.~¢ mmol) and BOP (302 mg, 0.6~ mmol) WO 97/2625a PCTrUS97/00572 were combined in 3 mL DMF. After 5 d the DMF was removed on a rotar~ evaporator, the residue was diluted with EtOAc, then wa,shed with water, .sat. NaHCO3, arld brine, dried ~MgSO4), filtered and concentrated. Flash chromatography (,silica, 25 % i-PrOH/EtOAc) 5 provided 3-7 a~ a colorless oil.
TLC Rf 0.30 (silica, 25 % i-PrOH/l~tOAc) 1 H-NMR (400 MHz, CDC13): ~ g.05 (d, J = 4 Hz, lH), 7.85 (d, J = 7 Hz, 2H), 7.57 (t, J = 7 Hz, lH), 7.55-7.45 (m, 2H), 7.42 (m, lH), 6.80 (brt, IH),6.54(dd,J=6,4Hz, lH),6.45(m, lH),6.39(d,J=8Hz, lH), :~.19 (m, lH), 4.16 (ABX dd, J = 17, 7 Hz, lH), 4.08-3.95 (m), 3.~5-3.75 (m, 2H), 3.29 (q, J = 6 Hz, 2H), 2.40-2.32 (m, 2H), 1.85 (m, J
- 7 Hz, 2H), 1.75 (m, 2H), 1.10 (t, J = 7 Hz, 3H).
o ~3,NH NH~l ~6CO2H
5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-,B-alanine trifluoroacetate salt (3-8) Ester 3-7 (59 mg, 0.12 rnrnol) was dissolved in 1 mL THF, then ] N LiOH (0.29 mL, 0.29 mmol) was added. After stirring overnight the reaction was concentrated, the mi~ture was concentrated.
Flash chromatography (silica, 22:20: 1: 1 EtOAc/EtOH/H2O/NH4OH), follo~,~ed by prep. HPLC ((:~-18, 0.1 % TFA/CH3CN/H20) and Iyophilization provided 3-8 as a white solid.
TLC Rf 0.26 (silica, 22:20: 1: 1 EtOAc/EtOH/H2O/NH4OH) lH-NMR (400 MHz, D2O): o 7.83-7.75 (m, 3H), 7.70 (d, J = 6 Hz, lH), 7.67 (d, J = 7 Hz, lH), 7.58 (t, J = 7 Hz, 2H), 6.96 (d, J = 9 Hz, lH), 6.80 (t, J = 7 Hz, lH), 3.86-3.80 (m, 3H), 3.55 (dd, J = 14, 4 Hz, lH), 3.36 (m, 2H), 3.29 (dd, J = 14, 8 Hz, IH), 2.39 (m, 2H), 1.72 (m, 4H).
WO 97/26250 PCT~US97/00572 BocNH ~' CO2H
N~
H-Sar-OEt-HCI
BOP, NMM
DMF
Me BocNH ~ ~N~,CO2Et N~ O
NaOH, EtOH ~N
BocNH ~ N~,CO2H ~~
N O H, 4-3 \ / H2N ~CO2Et \/ 4-4 BOP, NMM ,~ H
Me ~ H-BocNH N~JI' H CO2Et N~ O
W O 97/26250 PCTrUS97/00572 1) NaOH, EtOH /~N
2) TFA, CH2C1 Me O H-H2N ~ ~ N~Ll N ~CO2H
N O
Boc NH~CH3 1. n-BuLi/BrCH2CH2CO2Et N~ 2. NaOH, EtOH
Boc NH ~'~'~ CO2H
N
~:L
4-(2-N-Boc-aminopyridin-4-yl)butanoic acid (4-1~
The protected picoline ~90 g, 0.43 mol) was dissolved in 3 L THF under N2, cooled to -7~~C, and n-BuLi (1.6 M, 675 mL, 1.02 10 mol) was added during 30 min. The mixture was allowed to warm to RT for 1 h, then the resulting orange suspension was cooled to -78~C.
Methyl 3-bromopropionate (79 g, 0.47 mol) was added during 2 min.
After 15 min the cooling bath was removed and the mixture was allowed to wann to -20~C at which point it was quenched with 60 mL
15 HOAc in 250 mL THF. The solution was diluted with 2L EtOAc, washed with water, sat. NaHCO3, and brine, dried (MgSO4). The aqueou~ layers were re-extracted with EtOAc (2x), and these organic layers were filtered, concentrated, and dissolved in 1.5 L EtOH ~nd 1.5 L 1 N NaOH (1.5 mol). After I h the reaction was concentrated by 1/3, r diluted with 4 L EtOAc, the aqueous layer was removed. The pH of the 5 aqueous layer was adjusted to 4-5 with 10% KHSO4, then extracted with EtOAc (2 x 3L). The EtOAc layer~s were washed with brine, dried (MgSO4), filtered and concentrated7 providing 4-1 as a yellow oil.
Me BOCN H ~5~ ~ N~,CO2Et N O
4-(2-Boc-amino-pyridin-6-yl)butanoyl-.sarco~sine ethyl ester (4-2) Acid 4-1 (200 mg, 0.71 mmol), H-Sar-OEt-HCl (130 mg, 0.~S4 mmol), NMM (314 ,uL, 2.9 mmol) and BOP (37P, mg, 0.86 mmol) were combined in 5 mL DMF. After stirring overnight the reaction mixture was diluted with EtOAc, washed with water (5x), sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 50-70 % EtOAc/hexane) provided 4-2 as a colorless oil.
TLC Rf 0.54 (silica, 80% EtOAc/hexane) IH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major rotomer ~i 8.12 (d, J = 5 Hz, lH), 7.79 (s, lH), 7.48 (br s, lH), 6.83 (d, J = 6 Hz, lH), 4.19 (q, J = 7 Hz, 2H), 4.11 (s, 2H), 3.03 (s, 3H), 2.68 (t, J = 7 Hz, 2H), 2.39 (t, J = 7 Hz, 2H), 2.02 (qn, J = 7 Hz, 2H), 1.53 (s, 9H) 1.26 (t, J = 7 Hz, 3H).
W O 97126250 PCTrUS97/00572 = - 67 -Me BocNH~,~ ~~,N~,CO2H
N~ O
4-(2-Boc-amino-pvridin-4-vl)butanoyl-sarcosine (4-3) Ester 4-2 (91 mg, 0.24 mmol) was dissolved in 2.4 mL
EtOH, and 1 N NaOH (0.60 mL, 0.60 mmol) was added. After 45 min 5 the mixture was concentrated, then diluted with EtOAc, washes with 10 % KH[SO4 and brine, dried (MgSO4) filtered and reconcentrated, providing acid 4-3 as a gla,ss.
TLC Rf0.1~ (silica, 18:1:1, CH2C12/MeOH/HOAc) lH-NMR (400 MHz, CDC13): 1:1 mixture of amide rotomers ~ ~.03-7.~2 (m, 3H), 6.86 (br s, lH), 4.15/3.96 (s, 2H), 3.06/3.02 (s, 3H), 2.75-2.65 (m, 2H), 2.40 (m, 2H), 2.22-2.00 (m, 2~I), 1.53 (s, 9H).
/~\ H
~\~
Me O H~-BocNH ~ ~ N'JI' N ~,CO2Et N O
4-(2-E~oc-amino-pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-15 yl)ethyll-~-alanine ethyl ester (4-5) ~ Acid 4-3 (~4 mg, 0.24 mmol), amine 4-4 (see Duggan et al., U~S. Patent 5,264,420) (6~S mg, 0.26 mmol), NMM (104 ,uL, 0.95 ~ mmol) and BOP (127 mg, 0.29 mmol) were combined in 2.4 mL
CH3CN. After stirring overnight the mixture was diluted with EtOAc, 20 washed with water (4x), sat. NaHCO3, and brine, dried (MgSO4) and W O 97/26250 PCTrUS97/00572 concentrated. Flash chromatography (silica, EtOAc) provided 4-5 as a colorless oil TLC Rf 0.66 (silica, 20 % MeOH/EtOAc) lH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major S rotomer ~ 8.12 (s, lH), ~.09 (d, J = 5 Hz, lH), 7.7~s (s, lH), 7.55 (d, J =~S Hz, lH), 7.39 (s, lH), 7.34 (d, J = ~ Hz, IH), 7.17 (t, J = 8 Hz, lH), 7.08 (t, J = 7 Hz, lH), 7.03 (d, J = 2 Hz, lH), 6.79 (dd, J = 5, 1 Hz, lH), 6.71 (d, J = 9 Hz, lH), 4.32 (m, lH), 4.16-4.05 (m, 3H), 4.00 (AB d, J
= 15 Hz, lH), 3.94 (AB d, J = 15 Hz, lH), 3.04 (s, 3H), 2.77 (m, 2H), 2.63 (t, J = ~ Hz, 2H), 2.53 (m, 2H), 2.36 (m, 2H), 2.02-1.90 (m, 4H), 1.53 ~s, 9H), 1.22 (t, J = 7 Hz, 3H).
H2N ~ H
4-(2-Aminopyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl] -~3-alanine (4-6) Ester 4-5 (20 mg, 34 ~mol) was dissolved in 350 ,uL EtOH, then l N NaOH (P~5 }IL, g5 ,umol) was added. After 2 h the reaction was diluted with EtOAc, washed with 10 % KHSO4 and brine, dried (MgSO4), filtered and concentrated. The residue was dissolved in 1 mL
CH2Cl2, treated with 1 mL TPA for l h, then concentrated and azeotroped with toluene. Flash chromatography (silica, 50:1:1, EtOH/H20/NH40H) provided 4 6 as an off-white ~olid.
TLC Rf 0.55 (silica, 20: 1: l EtOH/H2O/NH4OH) lH-NMR (400 MHz, CD30D): 2:1 mixture of amide rotomers 7.72/7.67 (d, J = 6 Hz, lH), 7.52 (t, J = ~ Hz, lH), 7.30 (t, J = 8 Hz, lH), 7.07-6.90 (m, 4H), 6.55/6.54 (s, lH), 6.49 (s, lH), 4.36-4.25 (m, W O 97/26250 PCT~US97/00572 lH), ~.14-3.93 (m, 2H), 3.06/2.93 (s, 3H), 2.60 (t, J = R Hz, 2H), 2.55-2.45 ~m, 4H), 2.34 (t, J = 7 Hz, lH), 2.05-1.84 (m).
W 097/26250 PCT~US97/00572 H2N ~CH3 Boc20 ~ CICH2CH2CI, BocN H '[~,C H3 2 ~N - SO
1 ) LDA, THr, -23~C
2) allyl bromide, -78~C Cbz-Gly-OH
BOP, NMM
BocNH ~ N~ DMF
~ O
CbzNH ~ CO2-t-Bu 1) 9-BBN, THF ' N~ /=\
~ , 2) H202, NaOH H H HN-S0 BocNH~ N~ ,OH 5-6 ~1 1 H2, 10 % Pd/C
5-4 EtOAc Jones Reagent ~ acetone O
BocNH '¢~~ CO2H H2N~ N~CO2-t-B~
5-5 \~/ 5-7 BOP, NMM
, DMF
W O 97;26250 PCTnUS97./00572 SCHEME S CONTINUED
H O
BocN H N~ /~ N ~ N ~CO2-t-Bu H H N- SO
TFA, CH2C12 H2N ~ ~ /~, N J~ N ~C02H
BocNH~,CH3 2-(Boc-amino)-6-methylpyridine (5-2) 2-Amino-6-picoline (5.0 g, 46.2 mmol) and Boc2O (11.1 g, 50.g mmol) were combined in 150 mL dichloroethane. After heating at reflux for 6 h, additional Boc20 (2.0 g, 9.2 mmol) was added, and the 10 reaction was heated overnight. After concentration, the reaction mixture was flash filtered (silica, CH2C12), providing 5-2 as a waxy solid.
TLC Rf 0.21 (~ilica, CH2cl2) lH-NMR (400 MHz, CDC13): ~ 7.70 (d, J = 8 Hz, IH), 7.54 (t, J = 8 Hz, lH), 7.19 (br s, lH), 6.X0 (d, J = 7 Hz, lH), 2.42 ~s, 3H), 1.51 (.s, 9H).
W 097/26250 PCTrUS97/00572 BocNH ~ N~
2-Boc-amino-6-(4-butenyl)pyridine (5-3) Methylpyridine 5-2 (4.0 g, 19.2 mmol) was dissolved in 40 mL THF, cooled to -23~C, and LDA (2 M, 24 mL, 4~s mmol) wa,~ added S dropwise. After 30 min the mixture was cooled to -78~C and allyl bromide (2.49 mL, 2.g~ mmol) wa~ added dropwise. After 15 min more, the reaction was quenched with sat. NH4CI, walTned to RT, diluted with EtOAc, and the organic layer wa,s washed with brine.
After drying (MgSO4), filtration and concentration, flash 10 chromatography provided 5-3 as a yellow oil.
TLC Rf 0.40 (silica, 75 % CH2C12/hexane) I H-NMR (300 MHz, CDC13): ~ 7.72 (d, J = 8 Hz, I H), 7.56 (t, J = ~ Hz, lH), 7.16 (br s, lH), 6.80 (d, J = 7 Hz, lH), 5.~5 (m, lH), 5.03 (dm, J =
17 Hz, lH), 4.97 (dm, J = 10 Hz, lH), 2.74 (t, J = 7 Hz, 2H), 2.42 (~lm, 1~ ~ = 7 Hz, 2H), 1.52 (~, 9H).
BocNH ~¢~ ~ OH
2-(Boc-amino)-6-(4-hydroxybutyl)pyridine (5-4) A ,~;olution of aUcene 5-3 (55~s mg, 2.25 mmol) in 2 mL
20 THF was added dropwise to a ,solution of 9-BBN (0.5 M in THF, 4.95 ~nL, 2.4~s mrnol). After stirring overnight, and additional portion of 9-BBN (0.5 M, 1.1 mL, 0.55 mrnol) wa~ added and the reaction wa.s continued I h more. The reaction was quenched by the successive addition of EtOH (1.5 mL), 6 N NaOH (0.5 mL), and 30 % H2~2 (1.0 25 mL, exothermic), and heating to 50~C for lh. The cooled mixture wa~
~aturated with K2C03, then partitioned between EtOAc and water. The aqueous phase wa~ reextracted with EtOAc, the combined organic W O 97f26250 PCTAUS97/00572 phases were washed with brine, dried (MgSO4), filtered and - concentrated. Flash chromatography (silica, 40 % EtOAc/hexane) provided alcohol 5-4 as a colorless oil.
TLC Rf 0.26 (silica, 40 % EtOAc/hexane) IH-NMR (400 MHz, CDC13): ~ 7.73 (d, J = 8 Hz, lH), 7.56 (t, J = 8 Hz, lH), 7.20 (br s, lH), 6.~S0 (d, J = 7 Hz, lH), 3.67 (t, J = 7 Hz, 2H), 2.70 (t, J -= 7 Hz, 2H), 1.77 (qn, J = 7 Hz, 2H), 1.61 (m, 2H), 1.51 (s, 9H).
BocNH ~~ CO2H
4-(2-Boc-aminopvridin-6-yl)butanoic acid (5-5) A solution of alcohol 5-4 (247 mg, 0.93 mmol) in 5 mL
acetone was cooled to 0~C and a solution of Jones Reagent was added drop~ise. As the color of l:he reaction changed from brown to green, additional Jones ~eagent was added, until the alcohol wa~ no longer detected b~y TLC (3.5 h). After quenching with i-PrOH the mixture was diluted with EtOAc, washed with 5 % KHSO4 and brine, dried (MgSO4), filtered and concentrated, providing 5-5 as an of~-white waxy solid.
IH-NMR (400 MHz, CDC13): ~i 9.13 (br s, lH), 7.90 (d, J = ~s Hz, lH), 7.64 (t, J = 8 Hz, lH), 6.85 (d, J = 8 Hz, lH), 2.80 (t, J = 8 Hz, 2H), 2.46 I(t~ J = 7 Hz, 2H), 2.01 (qn, J = 7 Hz, 2H), 1.54 (s, 9H).
CbzNtl ~,1~ N~CO2-t-Bu H H NH-N-C~bz- Iycyl-2(S)-phenylsulfonamido-~3-alanine t-butyl ester ~5-6) Amine 1 5 (0.42 g, 1.25 mmol), Cbz-Gly-OH (288 mg, 1.38 mmol), NMM (0.55 mL, 5.0 mmol) and BOP (829 mg, 1.8~$
W 097/26250 PCT~US97/00572 mmol) were combined in 6 mL DMF. ~fter stirring overnight the solvent was evaporated, the residue wa~ taken up in EtOAc, the organic solution was washed with water (2x), 5 % KHS04, sat. NaHC03 and brine, dried (MgS04), fi}tered and concentrated. Flash chromatography S (.~ilica, 60 % EtOAc~exane) provided 5-6 a~ a white gla,ss.
TLC Rf 0.27 (silica, 60 % EtOAc/hexane) H-NMR (400 MHz, CDC13): ~ 7.~3 (d, J = 7 Hz, 2H), 7.58 (t, J = 7 Hz, lH), 7.50 (t, J = 8 Hz, 2H), 7.42-7.30 (m, SH), 6.55 (br s, lH), 5.59 (d, 3 = 7 Hz, lH), 5.40 (br s, lH), 5.16 (~, 2H), 3.95-3.70 (m, 4H), 3.34 (m, l O l H), l .27 (.~, 9H).
H2N ~ N ~CO2-t-Bu H H N-S02~
Glycyl-2(S)-phenylsulfonamido-,B-alanine t-butyl e,ster (5-7) A solution of 5-6 (0.54 g, l.10 mmol) in l 1 mL EtOAc was treated with lO % Pd/C (108 mg) and stirred under a H2 balloon overnight. After addition of more 10% Pd/C (lOO mg) and hydrogenation for 5 d the mixture wa~ filtered through Celite and concentrated, providing 5-6 as a white glas.~i.
lH-NMR (400 MHz, CD30D): ~ 7.~4 (dm, J = ~ Hz, 2H), 7.61 (tm, J =
TFA ~H2N ~,1 1-7 NHSO2Ph Glycyl-2(S)-phenyl~ulfonamido-,B-alanine t-butyl ester trifluoroacetate 20 salt (1-7) Protected amide 1-6 (576 mg, 1.26 mmol) was dissolved in 6.3 mL CH2C12, cooled to -15~C, and TFA (6.3 mL) was added. After 25 min the reaction was concentrated, providin~ amine 1 7.
TLC Rf 0.36 (silica, 9:1:1 CH2C12/MeOH/HOAc).
W O 97/2625~ - - PCTr~S97/00572 H2N~<,N~NJ~ N ~CO2-t-Bu~
4-(2-Aminothiazol -4-yl)butanoyl-glycyl-2(S)-phenyl.sulfonamido-,B-alanine t-butyl ester (1 -g~
Acid ~-4 (300 mg, 1.35 mmol), amine 1 7 (600 mg, 1.37 5 mmol), HOBT (219 mg, 1.14 mmol) and NMM (445 ,uL, 4.04 mmol) were combined in 13 mL DMF, cooled to -15~C, and EDC (310 mg, 1.61 mmol) was added. The reaction was warmed to RT, stirred ovemight, then diluted with LtOAc, washed with water, sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash 1~ chromatography (silica, 20% MeOH/EtOAc) provided 1 ~ as yellow solid.
TLC Rf 0.55 (silica, 20% MeOH/EtOAc) 1H-r~MR (400 MHz, CDC13): ~ 7.80 (d, J = 7 Hz, 2H), 7.55 (ABX t, J =
7 Hz, lH), 7.47 (ABX t, J = 8 Hz, 2H), 7.35 (br s, lH), 7.04 (br m, lH), 6.12 (s, lH), 5.41 (br s, 2H), 4.05-3.95 (m, 3H), 3.69 (m, IH), 3.39 (ddd, lH), 2.70-2.55 (m, 2H), 2.33 (m, 2H), 2.01 (qn, J = 7 Hz, 2E~), 1.27 (~;, 9~) H2N y~N~ N N ~~2H
S C) H H ~ S0 4-(2-Aminothiazol-4-yl)butanoyl-glycyl -2(S)-phenylsulfonamido-,B-alaninç (1 -9) ~- Ester 1 ~s ~365 mg, 0.69 mmol) was dissolved in CH2cl2 (3.5 mL), then TFA (3.5 mL) was added. After 5 h the reaction 25 mixture was concentrated, azeotroped with toluene, then purified by sequential flash chromatography (silica, 22:20: 1: 1 W O 97/26250 PCT~US97/00572 EtOAc/EtOH/H20/NH40H, then silica, 4:1:1 CH2(~12/MeOH/HOAc, then 7:1:1 CH2cl2lMeoHlHoAc)~ providing 1 9 as a white solid.
TLC Rf 0.33 (silica, 7:1:1 CHzC12/MeOH/HOAc) 1 H-NMR (400 MHz, CD30D): ~ 7.~s6 (d, J - 7 Hz, 2H), 7.5~ (ABX t, J
= 7 Hz, lH), 7.52 (ABX t, J = 8 Hz, 2H), 6.27 (s, lH), 3.~,9 (AB d, J =
17 Hz, lH), 3.77 (AB d, J = 17 Hz, lH), 3.64 (t, J = 6 Hz, lH), 3.53 (AB dd, lH), 3.41 (AB dd, lH), 2.57 (t, J = 7 Hz, 2H), 2.35-2.25 ~m, 2H~, 1.95 (m, 2H).
L-asparagine NaOH, H20, PhSO2CI
H2NC--~
H NHSO2Ph NaOH, dioxane, Br2 0~C to 90~C
-.. H2 \1 ~,CO2H
H N HSO2Ph - 1-2a a. isobutylene, H2SO4, dioxane / SOCI2 \b. 1N HCI/ether / ethanol HC~ ~ H2N ~6 2 ~~
H NHSO2Ph H NHSO2Ph N-Phenylsulfony~-L-a~paragine ( 1 - l a) To a stirred solution of L-asparagine (Aldrich) (lO g, 76 mmol), NaOH (3.4 g, 85 mmol), H20 (50 mL), and dioxane (50 mL) at 0~C was added PhS02CI (10.6 mL, ~4 mrnol). After 1 min, NaOH (3.4 g) in ~2~ (50 mL) was added and the reaction mixfure stirred for 30 min. The reaction mixture was then concentrated to remove the dioxame then washed with EtOAc. ~he aqueous phase was then cooled to 0~C and acidified to pH 5.0 with conc. HCI to effect product precipitation. The re,sulting solid was collected by filtration, washed with II20 (20 mL) and dried at 50~C under vacuum to give N-phenylsulfonyl-L-asparagine (I-la) as a white solid.
~f 0.40 (silica, l O: 1:1 ethanol/H20/NH40H). I H NMR (300 MHz, D20) ~ 7.59 (m, 2H), 7.26 (m, 3H), 3.92 (m, lH), 3.02 (m, lH), 2.35 (m, l]H).
W 097/26250 PCT~US97100572 H2N ~6 C~2H
H N HSO2P h 3-Amino-2(S)-phenylsulfon~laminopropionic acid (1-2b) To stirred solution of NaOH (15.6 g, 0.4 mol) in H2O (70 mL)1 cooled with an icebath, was added bromine (3.6 mL, 0.07 mol) dropwise. After 5 min, a cold .solution of N-phenylsulfonyl-L-asparagine, l-la (14.6 g,54 mmol) and NaOH (4.3 g, 0.1 mol) in H2O
(50 mL) was added in one portion. The solution was stirred for 20 min at 0~C then 30 min at 90~C. The reaction mixture was recooled to 0~C, and the pH adjusted to 7 through dropwise addition of conc. HCl. The 10 white precipitate forrned was collected by filtration and then dried to give ~) as a white solid. lEI NMR (300 MHz, D2O) ~ 8.00, 7.50 (m, 5H), 3.8~s (m, lH), 3.37 (m, lH), 3.12 (m, lH).
2 ~6 H NHSOZPh 1~ Ethyl 3-Amino-2(S)-phenylsulfonylaminopropionate hydrochloride (3-4) Arnino acid 1 -2a ( 1.0 g, 4.1 mmol) was suspended in 20 mL EtOH, cooled to 0~C, and SOC12 (1.5 mL, 21 rnrnol) was added dropwise. After stirring at RT overnight the mixture was concentrated, 20 triturated with Et2O (2x), and dried, providing 3-4 (1.26 g) as a hygroscopic yellow solid.
lH-NMR (300 MHz, d6-DMSO): ~ ~.30 (br s), 7.79 (d, J = ~ Hz, 2H), 7.70-7.60 (m, 3H), 4.21 (t, J Hz, lH), 3.90-3.~0 (m, 2H), 3.09 ~ABX
dd, J = 13, 6 Hz, lH), 2.90 (ABX dd, J = 13, ~ Hz, 2H), 0.97 (t, J = 7 2~ Hz, 3H).
W O 971262S0 PCT~US97/00~72 HCI- H2N~
H NHSO2Ph tert-Butyl 3-Amino-2(S)-phenylsulfonylaminopropionate hydro-chloride f 1-~) In a Fischer-Porter tube, a mixture of 1-2a (10.2 g, 42 mmol) and DME~ (150 mL) was sequentially treated with ~2S04 (6.4 mL, ~.12 mol), cooled to -7~~C, and then condensed isobutylene (75 mL). The cooling bath was removed. After 24 h, ice/water (250 mL) was added ~ollowed by washing with ether (2x). The a~ueous phase was basified with aq 6N NaOH, then saturated with NaCI, followed by extraction with EtOAc (3x). The combined extracts were washed with brine, dried (MgS04), and concentrated to give a white .solid. This was dissolved in CH2C12 and treated with lN HCl/ether (22 mL), and then concentrated to give 1 5 as a glassy yellow solid.
lH NMR (400 MHz, DM~O) o 8.25-8.00 (m, 4H), 7.85-7.5fs (m, 5H), 4.0~ (m, lH), 3.10 (m, IH), 2.73 (m, lH), 1.17 (s, 9H).
W O 97126250 PCT~U~97/00572 ~, HCI-H2Ny~ ~CO2H O J
1-4 HCI-H2N ~J~ ~ CO2Me ~ / 2-1 EDC, HOBT, NMM
DMF, -1 5~C to RT ~
y ~ N ~J~ N ~CO2Me 1 NNaOH
MeOH ¢~
TFA H2Ny~N~ ~ H H ~
S O
~ A
W O 97/2625~ PCTMS97/00572 . - 57 -~ H (CH2)2Ph NHJlNH~ C02CH3 S O
4-(2-Aminothiazol -4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl)-~3-alanine methyl ester (2-2) Acid 1 4 (300 mg~ 1.35 mmol), amine 2-1 (405 mg, 1.35 mmol) (prepared as described in Duggan et al., U.s. Patent 5,264,~20) HOBT (219 mg, 1.62 mmol) and NMM (445 ~L, 4.04 mmol) were combined in 7 mL DMF, cooled to -15~C, and EDC (310 mg, 1.61 mmol) was added. The reaction was warmed to RT, stirred overnight, 10 then cliluted with EtOAc, washed with water, ~sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography ~ilic.l, 10% MeOH/EtOAc) provided 2-2 as a yellow oil.
TLC Rf 0.32 (silica, 10 % MeOH/EtOAc) 1 H-NMR (400 MHz, CDC13): o 7.82 (d, J = 7 Hz, 1 H), 7.65 (d, ~ = ~
1~ Hz, lH), 7.40-7.10 (m, 5H), 6.93 (d, J = 8 Hz, lH), 6.10 (s, lH), 4.31 ~m, IH), 3.96 (ABX dd, J = 17, 6 Hz, lH), 3.89 (ABX dd, J = 17, 5 Hz, lH), 3.64 (s, 3H), 2.68-2.54 (m), 2.32-2.17 (m, 2H), 2.25~ 0 (m).
~ H (CH2)2Ph ~NHJlNH~CO2H
S O
~ 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-~-alanine trifluoroacetate salt ~2-3) Ester 2-2 (220 mg, 0.51 mmol) and 1 N NaOH (1.3 mL, 1.3 mmol) were combined in 5 mL MeOH. After 3 d the reaction 25 mixture was concentrated, purified by flash chromatography (.~iilica, 9: 1: 1 CH2C12/MeOH/HOAc), then preparative HPLC (Cl g, 0.1 % TFA
W O 97/26250 PCTAUS97/00~72 - 5~ -in CH3CN/H20), providing, after Iyophilization, acid 2-3 a~ a white ~olid.
TLC Rf 0.54 (silica, 4:1:1 CH2C12/MeOH/HOAc) lH-NMR (400 MHz, CD30D): ~ 7.26-7.10 ~m, 5H), 6.52 (~, lH), 4.23 5 (m, lH), 3.~¢ (AB d, J = 17 Hz, IH), 3.79 (AB d, J = 17 Hz, lH), 2.72-2.5~ (m, 4H), 2.51 (d, J = 7 Hz, 2H), 2.34 (t, J = 7 Hz, 2H), 1.99-1.75 (m, 4H).
W O 97/262~,0 PCTAUS97/00572 ~, NH~ ~N SO~
Cbz-G Iy-OH
NaH, DMF, 0~C to RT; isobutyl chlorofor,nate, Ethyl 5-bromopentanoate NMM
~ RT to 75~C EtOAc, -15~C to RT
¢~N CO2E~ ~H
1 N NaOH, EtOH H2, 10 % Pd/C
EtOH
~, N~ N ~ CO2H ~
~J H2N J~ N ~CO2Et 3 3 \/ H H NH-SO
BOP, NMM
DMF
r W O 97/26250 PCTrUS97/00572 - 60 - v H H (~
N~ N ~ N~ N ~CO2Et oH H N - S0 LiOH, THF
TFA~ N'J~N ~C02H
NI~ ~ co2Et Ethyl 5-~2-pyridylamino)pentanoate (3-2) 2-Aminopyridine (3-1, 1.97 ~, 20.9 mmol) in 10 mL DMF
was added to a suspension of NaH (60 % in oil, 1.00 g, 25 mmol) in 80 mL DMF cooled to 0~C. After warming to RT for 45 min, ethyl 5-10 bromopentanoate (4.2 mL, 25 rnmol) wa~; added dropwi,se. Thismixture was heated at 75~C overnight, then cooled to RT, diluted with EtOAc, washed with water (2x), sat. NaHCO3, and ~rine, drie~
~MgSO4), filtered and concentrated. Flash chromatography (silica, 50%
then 70 % EtOAc/hexane) provided 3-2 as a yellow oil.
15 TLC R~ 0.55 (silica, 70 % EtOAc/hexane) H-NMR (400 MHz, CDC13): ~ ~.07 (dd, J = 5, I Hz, lH), 7.40 (m, lH), 6.55 (m, lH), 6.37 (d, J = ~ Hz, lH), 4.4~ (br s, lH), 4.13 (4, J = 7 Hz, 2H), 3.29 (q, J = 7 Hz, 2H), 2.35 (t~ J = 7 Hz, 2H), l.g0-1.55 (m 4H), 1.25 (t, J = 7 Hz, 3H).
W O 97126250 PCT~US97100572 ~,NH'--~--C02H
5-(2-Pyridylamino)pentanoic acid (3-3) Ester 3-2 (0.41 g, 1.~4 mmol) was dissolved in 18 mL
5 EtOH, 1 N NaOH (4.6 mL, 4.6 mmol) was added, and the reaction was stirrecl overnight. The pH of the solution was adjusted to 7 with I N
HCl, and concentration provided a white solid containing acid 3-3 and NaCI.
TLC Rf 0.06 (silica, l 9:1:1 CH2Cl2/MeOH/HOAc) lH-NMR (400 MHz, D2O): ~ 7.~1 (m, lH), 7.77 (d, J = 6 Hz, lH), 6.96 (d, J -- 9 Hz, lH), 6.~2 (t, J - 7 Hz, lH), 3.36 (t, J = 7 Hz, 2H), 2.24 (m, 2H), ~.72-1.50(m, 4H).
CbZNH~ NH~6co2Et !L5 N-Cb%-~lycyl-2(S~-phenylsulfonamido-,B-alanine ethvl ester (3-5) N-Cbz-glycine (339 mg, 1.62 mrnol) was dissolved in 8 mL
EtOAc, cooled to -15~C, then NMM (196 }IL, 1.~ mmol) and isobutyl chloroformate (230 ~lL, 1.8 mmol) were added. After 20 min, the 20 mixed anhydride solution was added to amine 3-4 (0.50 mg, 1.6 mmol) suspended in 5 mL EtOAc and the reaction was warrned to RT for 90 min. Following dilution with EtOAc, the mixture was washed with water, sat. NaHCO3, 5% KHSO4, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 75 % EtOAc/hexane) 25 provided amide 3-5 as a colorless oil.
TL~ ]R~ 0.29 (silica, 75 % EtOAc/hexane) W O 97/26250 PCTrUS97/00572 lH-NMR (300 MHz, CDC13): ~ 7.65-7.45 (m, 3H~, 7.40-7.25 (m, 5H), 6.6~ (t, J = 6 Hz, lH), 5.~s3 (d, J = ~ Hz, lH), 5.49 (t, J = 6 Hz, lH), 5.15 (.s, 2H), 4.04-3.95 (m, 3H), 3.~9-3.~5 (m, 2H), 3.73 (m, IH), 3.46 (m, lH), 1.11 (t, J = 7 Hz, 3H).
s O
H2N~NH ~CO2Et H- NHSO2Ph Glycyl-2(S)-phenyl.sulfonamido-~-alanine ethyl ester (3-6) Protected amine 3-5 (0.47 g, 1.01 mmol) was dissolved in 10 mL EtOH, 10 % Pd/C (94 mg) was added, and the reaction was stirred under an H2 balloon. After 4 h, additional 10 % Pd/C was added (94 mg), and the reaction was continued for 3 d. The mixture was filtered through Celite, concentrated, and azeotroped with CHC13, providing amine 3-6 as a gum.
l~-NMR (300 MHz, CDC13): ~ 7.95 (m), 7.~6 (d, J = 7 Hz, 2H), 7.60-7.45 (m, 3H), 4.05 (dd, J = 5, 6 Hz lH), 3.96 (q, J = 7 Hz, 2~), 3.g0-3.55 (m), 1.07 (t, J = 7 Hz, 3H).
[~,N H ~ ~6N HSO2Ph ~3-7 5 -(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine ethyl ester (3-7) Acid 3-3 (1~6 mg, 0.55 mmol), amine 3-6 (150 mg, 0.46 mmol), NMM (0.20 mL, 1.~¢ mmol) and BOP (302 mg, 0.6~ mmol) WO 97/2625a PCTrUS97/00572 were combined in 3 mL DMF. After 5 d the DMF was removed on a rotar~ evaporator, the residue was diluted with EtOAc, then wa,shed with water, .sat. NaHCO3, arld brine, dried ~MgSO4), filtered and concentrated. Flash chromatography (,silica, 25 % i-PrOH/EtOAc) 5 provided 3-7 a~ a colorless oil.
TLC Rf 0.30 (silica, 25 % i-PrOH/l~tOAc) 1 H-NMR (400 MHz, CDC13): ~ g.05 (d, J = 4 Hz, lH), 7.85 (d, J = 7 Hz, 2H), 7.57 (t, J = 7 Hz, lH), 7.55-7.45 (m, 2H), 7.42 (m, lH), 6.80 (brt, IH),6.54(dd,J=6,4Hz, lH),6.45(m, lH),6.39(d,J=8Hz, lH), :~.19 (m, lH), 4.16 (ABX dd, J = 17, 7 Hz, lH), 4.08-3.95 (m), 3.~5-3.75 (m, 2H), 3.29 (q, J = 6 Hz, 2H), 2.40-2.32 (m, 2H), 1.85 (m, J
- 7 Hz, 2H), 1.75 (m, 2H), 1.10 (t, J = 7 Hz, 3H).
o ~3,NH NH~l ~6CO2H
5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-,B-alanine trifluoroacetate salt (3-8) Ester 3-7 (59 mg, 0.12 rnrnol) was dissolved in 1 mL THF, then ] N LiOH (0.29 mL, 0.29 mmol) was added. After stirring overnight the reaction was concentrated, the mi~ture was concentrated.
Flash chromatography (silica, 22:20: 1: 1 EtOAc/EtOH/H2O/NH4OH), follo~,~ed by prep. HPLC ((:~-18, 0.1 % TFA/CH3CN/H20) and Iyophilization provided 3-8 as a white solid.
TLC Rf 0.26 (silica, 22:20: 1: 1 EtOAc/EtOH/H2O/NH4OH) lH-NMR (400 MHz, D2O): o 7.83-7.75 (m, 3H), 7.70 (d, J = 6 Hz, lH), 7.67 (d, J = 7 Hz, lH), 7.58 (t, J = 7 Hz, 2H), 6.96 (d, J = 9 Hz, lH), 6.80 (t, J = 7 Hz, lH), 3.86-3.80 (m, 3H), 3.55 (dd, J = 14, 4 Hz, lH), 3.36 (m, 2H), 3.29 (dd, J = 14, 8 Hz, IH), 2.39 (m, 2H), 1.72 (m, 4H).
WO 97/26250 PCT~US97/00572 BocNH ~' CO2H
N~
H-Sar-OEt-HCI
BOP, NMM
DMF
Me BocNH ~ ~N~,CO2Et N~ O
NaOH, EtOH ~N
BocNH ~ N~,CO2H ~~
N O H, 4-3 \ / H2N ~CO2Et \/ 4-4 BOP, NMM ,~ H
Me ~ H-BocNH N~JI' H CO2Et N~ O
W O 97/26250 PCTrUS97/00572 1) NaOH, EtOH /~N
2) TFA, CH2C1 Me O H-H2N ~ ~ N~Ll N ~CO2H
N O
Boc NH~CH3 1. n-BuLi/BrCH2CH2CO2Et N~ 2. NaOH, EtOH
Boc NH ~'~'~ CO2H
N
~:L
4-(2-N-Boc-aminopyridin-4-yl)butanoic acid (4-1~
The protected picoline ~90 g, 0.43 mol) was dissolved in 3 L THF under N2, cooled to -7~~C, and n-BuLi (1.6 M, 675 mL, 1.02 10 mol) was added during 30 min. The mixture was allowed to warm to RT for 1 h, then the resulting orange suspension was cooled to -78~C.
Methyl 3-bromopropionate (79 g, 0.47 mol) was added during 2 min.
After 15 min the cooling bath was removed and the mixture was allowed to wann to -20~C at which point it was quenched with 60 mL
15 HOAc in 250 mL THF. The solution was diluted with 2L EtOAc, washed with water, sat. NaHCO3, and brine, dried (MgSO4). The aqueou~ layers were re-extracted with EtOAc (2x), and these organic layers were filtered, concentrated, and dissolved in 1.5 L EtOH ~nd 1.5 L 1 N NaOH (1.5 mol). After I h the reaction was concentrated by 1/3, r diluted with 4 L EtOAc, the aqueous layer was removed. The pH of the 5 aqueous layer was adjusted to 4-5 with 10% KHSO4, then extracted with EtOAc (2 x 3L). The EtOAc layer~s were washed with brine, dried (MgSO4), filtered and concentrated7 providing 4-1 as a yellow oil.
Me BOCN H ~5~ ~ N~,CO2Et N O
4-(2-Boc-amino-pyridin-6-yl)butanoyl-.sarco~sine ethyl ester (4-2) Acid 4-1 (200 mg, 0.71 mmol), H-Sar-OEt-HCl (130 mg, 0.~S4 mmol), NMM (314 ,uL, 2.9 mmol) and BOP (37P, mg, 0.86 mmol) were combined in 5 mL DMF. After stirring overnight the reaction mixture was diluted with EtOAc, washed with water (5x), sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 50-70 % EtOAc/hexane) provided 4-2 as a colorless oil.
TLC Rf 0.54 (silica, 80% EtOAc/hexane) IH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major rotomer ~i 8.12 (d, J = 5 Hz, lH), 7.79 (s, lH), 7.48 (br s, lH), 6.83 (d, J = 6 Hz, lH), 4.19 (q, J = 7 Hz, 2H), 4.11 (s, 2H), 3.03 (s, 3H), 2.68 (t, J = 7 Hz, 2H), 2.39 (t, J = 7 Hz, 2H), 2.02 (qn, J = 7 Hz, 2H), 1.53 (s, 9H) 1.26 (t, J = 7 Hz, 3H).
W O 97126250 PCTrUS97/00572 = - 67 -Me BocNH~,~ ~~,N~,CO2H
N~ O
4-(2-Boc-amino-pvridin-4-vl)butanoyl-sarcosine (4-3) Ester 4-2 (91 mg, 0.24 mmol) was dissolved in 2.4 mL
EtOH, and 1 N NaOH (0.60 mL, 0.60 mmol) was added. After 45 min 5 the mixture was concentrated, then diluted with EtOAc, washes with 10 % KH[SO4 and brine, dried (MgSO4) filtered and reconcentrated, providing acid 4-3 as a gla,ss.
TLC Rf0.1~ (silica, 18:1:1, CH2C12/MeOH/HOAc) lH-NMR (400 MHz, CDC13): 1:1 mixture of amide rotomers ~ ~.03-7.~2 (m, 3H), 6.86 (br s, lH), 4.15/3.96 (s, 2H), 3.06/3.02 (s, 3H), 2.75-2.65 (m, 2H), 2.40 (m, 2H), 2.22-2.00 (m, 2~I), 1.53 (s, 9H).
/~\ H
~\~
Me O H~-BocNH ~ ~ N'JI' N ~,CO2Et N O
4-(2-E~oc-amino-pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-15 yl)ethyll-~-alanine ethyl ester (4-5) ~ Acid 4-3 (~4 mg, 0.24 mmol), amine 4-4 (see Duggan et al., U~S. Patent 5,264,420) (6~S mg, 0.26 mmol), NMM (104 ,uL, 0.95 ~ mmol) and BOP (127 mg, 0.29 mmol) were combined in 2.4 mL
CH3CN. After stirring overnight the mixture was diluted with EtOAc, 20 washed with water (4x), sat. NaHCO3, and brine, dried (MgSO4) and W O 97/26250 PCTrUS97/00572 concentrated. Flash chromatography (silica, EtOAc) provided 4-5 as a colorless oil TLC Rf 0.66 (silica, 20 % MeOH/EtOAc) lH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major S rotomer ~ 8.12 (s, lH), ~.09 (d, J = 5 Hz, lH), 7.7~s (s, lH), 7.55 (d, J =~S Hz, lH), 7.39 (s, lH), 7.34 (d, J = ~ Hz, IH), 7.17 (t, J = 8 Hz, lH), 7.08 (t, J = 7 Hz, lH), 7.03 (d, J = 2 Hz, lH), 6.79 (dd, J = 5, 1 Hz, lH), 6.71 (d, J = 9 Hz, lH), 4.32 (m, lH), 4.16-4.05 (m, 3H), 4.00 (AB d, J
= 15 Hz, lH), 3.94 (AB d, J = 15 Hz, lH), 3.04 (s, 3H), 2.77 (m, 2H), 2.63 (t, J = ~ Hz, 2H), 2.53 (m, 2H), 2.36 (m, 2H), 2.02-1.90 (m, 4H), 1.53 ~s, 9H), 1.22 (t, J = 7 Hz, 3H).
H2N ~ H
4-(2-Aminopyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl] -~3-alanine (4-6) Ester 4-5 (20 mg, 34 ~mol) was dissolved in 350 ,uL EtOH, then l N NaOH (P~5 }IL, g5 ,umol) was added. After 2 h the reaction was diluted with EtOAc, washed with 10 % KHSO4 and brine, dried (MgSO4), filtered and concentrated. The residue was dissolved in 1 mL
CH2Cl2, treated with 1 mL TPA for l h, then concentrated and azeotroped with toluene. Flash chromatography (silica, 50:1:1, EtOH/H20/NH40H) provided 4 6 as an off-white ~olid.
TLC Rf 0.55 (silica, 20: 1: l EtOH/H2O/NH4OH) lH-NMR (400 MHz, CD30D): 2:1 mixture of amide rotomers 7.72/7.67 (d, J = 6 Hz, lH), 7.52 (t, J = ~ Hz, lH), 7.30 (t, J = 8 Hz, lH), 7.07-6.90 (m, 4H), 6.55/6.54 (s, lH), 6.49 (s, lH), 4.36-4.25 (m, W O 97/26250 PCT~US97/00572 lH), ~.14-3.93 (m, 2H), 3.06/2.93 (s, 3H), 2.60 (t, J = R Hz, 2H), 2.55-2.45 ~m, 4H), 2.34 (t, J = 7 Hz, lH), 2.05-1.84 (m).
W 097/26250 PCT~US97/00572 H2N ~CH3 Boc20 ~ CICH2CH2CI, BocN H '[~,C H3 2 ~N - SO
1 ) LDA, THr, -23~C
2) allyl bromide, -78~C Cbz-Gly-OH
BOP, NMM
BocNH ~ N~ DMF
~ O
CbzNH ~ CO2-t-Bu 1) 9-BBN, THF ' N~ /=\
~ , 2) H202, NaOH H H HN-S0 BocNH~ N~ ,OH 5-6 ~1 1 H2, 10 % Pd/C
5-4 EtOAc Jones Reagent ~ acetone O
BocNH '¢~~ CO2H H2N~ N~CO2-t-B~
5-5 \~/ 5-7 BOP, NMM
, DMF
W O 97;26250 PCTnUS97./00572 SCHEME S CONTINUED
H O
BocN H N~ /~ N ~ N ~CO2-t-Bu H H N- SO
TFA, CH2C12 H2N ~ ~ /~, N J~ N ~C02H
BocNH~,CH3 2-(Boc-amino)-6-methylpyridine (5-2) 2-Amino-6-picoline (5.0 g, 46.2 mmol) and Boc2O (11.1 g, 50.g mmol) were combined in 150 mL dichloroethane. After heating at reflux for 6 h, additional Boc20 (2.0 g, 9.2 mmol) was added, and the 10 reaction was heated overnight. After concentration, the reaction mixture was flash filtered (silica, CH2C12), providing 5-2 as a waxy solid.
TLC Rf 0.21 (~ilica, CH2cl2) lH-NMR (400 MHz, CDC13): ~ 7.70 (d, J = 8 Hz, IH), 7.54 (t, J = 8 Hz, lH), 7.19 (br s, lH), 6.X0 (d, J = 7 Hz, lH), 2.42 ~s, 3H), 1.51 (.s, 9H).
W 097/26250 PCTrUS97/00572 BocNH ~ N~
2-Boc-amino-6-(4-butenyl)pyridine (5-3) Methylpyridine 5-2 (4.0 g, 19.2 mmol) was dissolved in 40 mL THF, cooled to -23~C, and LDA (2 M, 24 mL, 4~s mmol) wa,~ added S dropwise. After 30 min the mixture was cooled to -78~C and allyl bromide (2.49 mL, 2.g~ mmol) wa~ added dropwise. After 15 min more, the reaction was quenched with sat. NH4CI, walTned to RT, diluted with EtOAc, and the organic layer wa,s washed with brine.
After drying (MgSO4), filtration and concentration, flash 10 chromatography provided 5-3 as a yellow oil.
TLC Rf 0.40 (silica, 75 % CH2C12/hexane) I H-NMR (300 MHz, CDC13): ~ 7.72 (d, J = 8 Hz, I H), 7.56 (t, J = ~ Hz, lH), 7.16 (br s, lH), 6.80 (d, J = 7 Hz, lH), 5.~5 (m, lH), 5.03 (dm, J =
17 Hz, lH), 4.97 (dm, J = 10 Hz, lH), 2.74 (t, J = 7 Hz, 2H), 2.42 (~lm, 1~ ~ = 7 Hz, 2H), 1.52 (~, 9H).
BocNH ~¢~ ~ OH
2-(Boc-amino)-6-(4-hydroxybutyl)pyridine (5-4) A ,~;olution of aUcene 5-3 (55~s mg, 2.25 mmol) in 2 mL
20 THF was added dropwise to a ,solution of 9-BBN (0.5 M in THF, 4.95 ~nL, 2.4~s mrnol). After stirring overnight, and additional portion of 9-BBN (0.5 M, 1.1 mL, 0.55 mrnol) wa~ added and the reaction wa.s continued I h more. The reaction was quenched by the successive addition of EtOH (1.5 mL), 6 N NaOH (0.5 mL), and 30 % H2~2 (1.0 25 mL, exothermic), and heating to 50~C for lh. The cooled mixture wa~
~aturated with K2C03, then partitioned between EtOAc and water. The aqueous phase wa~ reextracted with EtOAc, the combined organic W O 97f26250 PCTAUS97/00572 phases were washed with brine, dried (MgSO4), filtered and - concentrated. Flash chromatography (silica, 40 % EtOAc/hexane) provided alcohol 5-4 as a colorless oil.
TLC Rf 0.26 (silica, 40 % EtOAc/hexane) IH-NMR (400 MHz, CDC13): ~ 7.73 (d, J = 8 Hz, lH), 7.56 (t, J = 8 Hz, lH), 7.20 (br s, lH), 6.~S0 (d, J = 7 Hz, lH), 3.67 (t, J = 7 Hz, 2H), 2.70 (t, J -= 7 Hz, 2H), 1.77 (qn, J = 7 Hz, 2H), 1.61 (m, 2H), 1.51 (s, 9H).
BocNH ~~ CO2H
4-(2-Boc-aminopvridin-6-yl)butanoic acid (5-5) A solution of alcohol 5-4 (247 mg, 0.93 mmol) in 5 mL
acetone was cooled to 0~C and a solution of Jones Reagent was added drop~ise. As the color of l:he reaction changed from brown to green, additional Jones ~eagent was added, until the alcohol wa~ no longer detected b~y TLC (3.5 h). After quenching with i-PrOH the mixture was diluted with EtOAc, washed with 5 % KHSO4 and brine, dried (MgSO4), filtered and concentrated, providing 5-5 as an of~-white waxy solid.
IH-NMR (400 MHz, CDC13): ~i 9.13 (br s, lH), 7.90 (d, J = ~s Hz, lH), 7.64 (t, J = 8 Hz, lH), 6.85 (d, J = 8 Hz, lH), 2.80 (t, J = 8 Hz, 2H), 2.46 I(t~ J = 7 Hz, 2H), 2.01 (qn, J = 7 Hz, 2H), 1.54 (s, 9H).
CbzNtl ~,1~ N~CO2-t-Bu H H NH-N-C~bz- Iycyl-2(S)-phenylsulfonamido-~3-alanine t-butyl ester ~5-6) Amine 1 5 (0.42 g, 1.25 mmol), Cbz-Gly-OH (288 mg, 1.38 mmol), NMM (0.55 mL, 5.0 mmol) and BOP (829 mg, 1.8~$
W 097/26250 PCT~US97/00572 mmol) were combined in 6 mL DMF. ~fter stirring overnight the solvent was evaporated, the residue wa~ taken up in EtOAc, the organic solution was washed with water (2x), 5 % KHS04, sat. NaHC03 and brine, dried (MgS04), fi}tered and concentrated. Flash chromatography S (.~ilica, 60 % EtOAc~exane) provided 5-6 a~ a white gla,ss.
TLC Rf 0.27 (silica, 60 % EtOAc/hexane) H-NMR (400 MHz, CDC13): ~ 7.~3 (d, J = 7 Hz, 2H), 7.58 (t, J = 7 Hz, lH), 7.50 (t, J = 8 Hz, 2H), 7.42-7.30 (m, SH), 6.55 (br s, lH), 5.59 (d, 3 = 7 Hz, lH), 5.40 (br s, lH), 5.16 (~, 2H), 3.95-3.70 (m, 4H), 3.34 (m, l O l H), l .27 (.~, 9H).
H2N ~ N ~CO2-t-Bu H H N-S02~
Glycyl-2(S)-phenylsulfonamido-,B-alanine t-butyl e,ster (5-7) A solution of 5-6 (0.54 g, l.10 mmol) in l 1 mL EtOAc was treated with lO % Pd/C (108 mg) and stirred under a H2 balloon overnight. After addition of more 10% Pd/C (lOO mg) and hydrogenation for 5 d the mixture wa~ filtered through Celite and concentrated, providing 5-6 as a white glas.~i.
lH-NMR (400 MHz, CD30D): ~ 7.~4 (dm, J = ~ Hz, 2H), 7.61 (tm, J =
8 Hz, l H), 7.54 (tm, J = ~ Hz, 2H), 4.00 (dd, J = 8, 5 Hz, 1 H), 3.59 (dd, J = 14, 5 Hz, lH), 3.37 (s, 2H), 1.25 (s, 9H).
BocN H N~ ~N ~ N ~CO2-t-Bu ~ o H H N-SO2~
4-(2-]3Oc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~3-alanirle t-butyl e~ster (5-Ps) Acid 5-5 (144 mg, 0.51 mmol), amine 5-7 (202 mg, 0.56 mmol), NMM (226 ,uL, 2.1 mmol) and BOP (241 mg, 0.77 mmol) were combined in 2.6 mL DMF. After stirring overnight the ~;olvent wa.s evaporated, the residue wa,s dissolved in EtOAc, washed with water, sat.
NaHCO3, 5 % KHSO4, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 10 % CHC13/EtOAc) provided S-g as an off-white glass.
TLC :Rf 0.22 (silica, 10 % CHC13/EtOAc) }H-NMR (400 MHz, d6-DMSO): ~ 9.53 (~, lH), 8.23 (br d, J = 7 Hz, lH), 7.92 (t, J = 6 Hz, lH), 7.91 (t, J = 6 Hz, lH), 7.76 (d, J = 7 Hz, 2H), 7.65-7.53 (m, 5H), 6.g7 (d, J = 7 Hz, lH), 3.~5 (br s, lH), 3.60 (t, J = 5 Hz, 2H), 3.20-3.10 (m, 2H), 2.60 (t, J = 7 Hz, 2H), 2.15 (t, J = 7 Hz, 2:H), l .85 (qn, J = 7 Hz, 2H), 1.46 (s, 9H), 1.1~ (s, 9H).
H2N N ' NJ~ N~CO2H
~'~ '~'~~ ~ H H H-S~2~
4-(2~ minopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-alanine (5-9) A solution of 5-8 (138 mg, 0.22 mrnol) in 1 mL CH2C12 was cooled to 0~C, treated with I mL TFA, and wa}med to RT for 5 h.
After concentration and azeotroping with toluene the residue was purified by fla.sh chromatography (.silica, 12:20: 1: 1, EtOAc/EtOH/H20/NH40H), providing 5-9 as a colorle~s glass.
W O 97/26250 PCT~US97/00572 TLC Rf 0.34 (silica,12:20: 1: 1, EtOAc/EtOH~H20~H40H) lH-NMR (400 MHz, D20): ~ 7.76 (dm, J = 7 Hz,2H),7.55-7.48 (m, 4H),6.69 (d, J - 7 Hz, lH),6.57 (d, J = ~ Hz, lH),3.72-3.62 (m,2H), 3.55 (dd, J = 8,5 Hz, lH),3.37 (dd, J = 13,8 Hz, lH),3.13 (dd, J = 13, 8 Hz, lH),2.63 (t, J = 7 Hz,2H),2.34 (t, J = 7 Hz,2H),1.96 ~qn, J = 7 Hz,2H).
W 09712625~ rCT~US97/00572 N~
1 0-5 H-Sar-OEt-HCI
BOP, NMM
DMF
Me "~ ~ N~,CO2Et N~ O
1 N NaOH ~,~
~tOH
Me H-,~ ~N~,CO2HH2N ~--C~2Et N~ O \
6-2 \/
BOP, NMM ~ H
DMF
Me ~ H-N~--U~ N ~CO2Et N O
-W O 97126250 PCTrUS97100572 - 7~S -/~\ H
/~ N
1 N NaOH
EtOH ~ J
Me ~ H-,~f ~N~JI' ~ CO2H
Me ~ ~ N~, CO2Et N~ O
4-(Pyridin-4-yl)butanoyl-sarcosine ethyl ester (6-1) 4-(4-Pyridyl)butanoic acid 10-5 (100 mg, 1.8 mmol), H-Sar-OEt-HCI (300 mg, 2.0 mmol), BOP (965 mg, 2.2 mmol~ and NMM
(700 IlL, 6.4 mmol) were combined in 9 mL DMF. After stirring 10 overnight the mixture wa.s diluted with EtOAc, washed with water (4x), sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated.
Flash chromatography (silica, 80% to 100 % Et~Ac/hexane) provided 6-1 as a cololless oil.
TLC Rf 0.44 (silica, 20 % MeOH/EtOAc) 1~ lH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major rotomer ~i 8.50 (d, J = 5 Hz, 2H), 7.14 (d, J = 5 Hz, 2H), 4.20 (~1, J = 7 Hz, 2H), 4.12 (s, 2H), 3.03 (s, 3H), 2.70 (t, J = 8 Hz, 2H), 2.39 (t, J = 7 Hz, 2H), 2.02 (~In, J = 7 Hz, 2H), 1.23 (t, J = 7 Hz, 3~I).
W O 97126250 PCTrUS97100~72 Me N~,CO2H
N ~ O
4-(Pyridin-4-yl)butanoyl-sarcosine (6-2) Ester 6-1 (324 mg, 1.22 mmol) wa~ dissolved in 6 mL
EtOH, then 1 N NaOH (2.4 mL, 2.4 mmol) wa.s added. After stirring overnight the mixture was concentrated, rediluted with EtOAc, extracted into 10 % KHSO4, then concentrated, providing acid 6-2, along with inorganic salts.
TLC l~f 0.16 (silica, 4: 1: 1 CH2C12/MeOH/HOAc) lH-NMR (400 MHz, CD30D): 1:1 mixture of amide rotomers, ~ g.41 10 (br s, 2H), 7.33 (m, 2H), 4.00/3.90 (s, 2~), 3.05/2.95 (s, 3H), 2.77-2.67 (m, 2]iI), 2.4~/2.37 (t, J = 7 Hz, 2H), 2.00-1.90 (m, 2H).
~=~
Me O H-~
N~ ~~ N'J~ N ~CO2Et 4-(Pyridin-4-yl)butanoyl-sarcosine-3 (R)-~2-(indol-3 -yl)ethyl]-,13-alanine 15 ethyl ester ~6-3) Acid 6-2 (2g~s mg, 1.22 mmol), amine 4-4 (318 mg, 1.22 mmol), BOP (647 mg, 1.5 mmol), and NMM (462 ~lL, 4.2 mrnol) were comb-ined in 6 mL DMF. After stirring overnight the mixture was diluted with EtOAc, washed with water (4x), sat. NaHCO3, and brine, 20 dried (MgSO4), filtered and concentrated. Flash chromatography (silica, EtOAc then 5 % MeOH/EtOAc) provided 6-3 as an orange oil.
TLC Rf 0.4 (20 % MeOH/EtOAc) W 097/26250 PCT~US97/OOS72 - ~0 -I H-NMR (400 MHz, CDC13): 4: 1 mixture of amide rotomers, major rotomer ~ 8.47 (br d, J = 5 H~, 2H), ~s.02 (br d, J = 6 Hz, 2H), 7.58 (dd, J = 16, g Hz, lH), 7.34 (dd, J = ~, 4 Hz, lH), 7.20-7.03 (m, 3H), 7.01 (s, IH), 6.72 (d, J = 9 Hz, lH), 4.33 (m, lH), 4.1 (t, J = 7 Hz, 3H), 3.9P, S (s, 2H), 3.05 (s, 3H), 2.90-2.45 (m), 2.39 (t, J = 7 Hz, 2H), 2.02-1.70 (m), 1.23 (t, J = 7 Hz, 3H)-/~\ H
/~
~ ~N--J~N~co2H
N O
4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-~2-(indol-3-yl)ethyl]-~-1 Q alanine (6-4) Ester 6-3 (400 mg, 0.~4 mmol) was dissolved in 4 mL
l~tOH, then 1 N NaOH (1.7 mL, 1.7 mmol) was added. After 90 min the reaction was neutralized with 1 N HCI (1.7 mL, 1.7 mmol) and concentrated to ~n oil. Flash chromatography (silica, 50: 1: 1 EtOH/H20/NH40H, then again with 12:10:1:1 EtOAc/EtOH/H20/NH40H) provided 6-4.
TLC Rf 0.17 (silica, 12:10:1:1 EtOAc/EtOH/H20/NE~40H) H-NMR (400 MH~, CD30D): 2:1 mixture of amide rotomers, ~ 8.3~S-~.2~ (m, 2H), 7.54-7.4~¢ (m, lH), 7.30-7.25 (m, 2H), 7.21-7.19 (m, lH), 2Q 7.07-6.92 (m, 3H), 4.36-4.27 (m, lH), 4.03-3.9~ (m, 2H), 3.06/2.93 (s, 3H), 2.~s6-2.60 (m, 4H), 2.52-2.32 (m), 2.05-l.g5 (m).
W O 97/26250 PCT~US97/OOS72 y Ethyl bromoacetate, TEA
CH2CI2, 0~C to RT
BocN H ~ C02H ~7 HN ~CO2Et BOP
NMM
DMF
BocNH~ ,N~,CO2Et N~ O
W O 97/262~0 PCT~US97/00572 SCHEME 7 (CONT'D) 1 N NaOH
MeOH
I~q BocNH ~N~,CO2H HCI-H2N ~CO2Et :Z~ \/ 7-S
BOP
NMM
DMF
~! ~
BocNH H~ CO2Et TFA, CH2CI2 W 097126250 PCTrUS97/00572 - ~3 -SCHEME 7 ~CONT'D) ,.
.
HCI-H2N ~ N~J~ N ~CO2Et LiOH, THF ¦
~7 ~ H -H2N~ N~J~ N~co2H
N ~7-8 HN ~,CO2Et N-Cyclopropyl~lvcine ethyl ester (7-2) Cyclopropylamine (12.1 mL, 175 mmol) and TEA (42 mL, 3~5 mmol) were combined at 0~C in 3~0 mL CH2C12, then ethyl 10 bromoacetate (19.4 mL, 175 mmol) was added dropwise. The reaction ~- wa~, warmed to RT for 3 h, then diluted with additional CH2C12, washed with water, .sat. NaHC03, and brine, dried (Na2S04), filtered and concentrated. Flash filtration (silica, 30 % EtOActhexane) provided 7-2 as a light yellow oil.
W O 97/26250 PCTnJS97/00572 - ~4 -TLC Rf 0.70 (~ilica, EtOAc) lH-NMR (400 MHz, CDC13): ~ 4.20 (q, J = 7 Hz, 2H), 3.45 (s, 2H), 2.23 (tt7 J = 6, 3 Hz, lH), 1.29 (t, J = 7 Hz, 3H), 0.43 (m, 2H), 0.36 (m, 2H).
y BocNH ~ N~CO2Et N~ O
4-(2-Boc-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycine ethyl ester (7-3) Acid 4 1 (0.86 g, 3.1 mmol), amine 7-2 (0.4~s g, 3.4 10 mmol), NMM (1.35 mL, 12.3 mrnol) and BOP (2.04 g, 4.61 mmol) were combined in 15 mL DMF. After stirring ovemight the mixture was concent~ated, redissolved in EtOAc, washed with water, 5 %
KHSO4, sat. NaHCO3 and brine, dried over MgSO4, filtered and concentrated. Flash chromatography (silica, 50 % EtC~Ac/hexane) 1~ provided 7-3 as a colorless oil.
TLC Rf 0.29 (silica, 50 % EtOAc~exane) lH-NMR (400 MHz, CDC13): ~ X.14 (d, J = S Hz, lH), 7.81 (.s, lH), 7.77 (br s, lH), 6.~4 (dd, J = 5, 1 Hz, lH), 4.18 (4, J = 7 Hz, 2H), 4.08 (s, 2H), 2.80 (tt, J = 7, 4 Hz, lH), 2.69 (t, J = 7 Hz, 2H), 2.60 (t, J=7 Hz, 20 2H)~ 2.02 (qn, J = 7 Hz, 2H), 1.53 (s, 9H), 1.27 (t, J = 7 Hz, 3H), 0.g3 (m, 2H), 0.72 (m, 2H).
W O 97126250 PCT~US97/00572 BocNH~ N~CO2H
N ~ O
4-(2-E~oc-amino-pyridin-4-yl~butanoyl-N-cyclopropyl~lycine (7-4) S Ester 7-3 (1.07 g, 2.64 mmol) wa~; dissolved in 26 mL
MeO~I, then treated with 1 N NaOH (6.6 m~, 6.6 rnrnol). After stirring overnight the reaction was concentrated, redissolved in water, the pH
was adjusted to 1 with 10 % KHSO4, then extracted with EtOAc (5x).
The a(~ueous pha.se was adjusted to pH 3 with a(l. NaOH, then reextr~cted with EtOAc (2x). The combined organic phases were washed with brine, dried (MgSO4), filtered and concentrated, providing 7-4 as a white foam.
TLC Rf 0.24 (silica, 19:1:1, CH2C12/MeOH/HOAc) lH-NMR (300 MHz, CDC13): ~ 9.15 (~r s, lH), 7.97 (d, J = 5 Hz, lH), 7.94 (:s, lH), 6.89 (dd, J = 5, 1 Hz, lH), 4.14 (,~, 2H), 2.81 (tt, J = 7, 3 Hz, lH),2.73(t,J=7Hz,2H),2.61 (t,J=7Hz,2H),2.04(qn,J=7 Hz, 2H), 1.51 (s, 9H), 0.85 (m, 2H), 0.76 (m, 2H).
W O 97i26250 PCTrUS97/00572 BocN H ~ N J~ N ~,CO2Et N O
4-(2-Boc-~mino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-13-alanine ethyl ester (7-6) Acid 7-4 (415 mg, 1.1 mmol), amine hydrochloride 7-5 (see procedure in EP 478 362 utilizing Boc-Gly(OEt) as starting material) (2~4 mg, 1.1 mmol), NMM (0.48 mL, 4.4 mmol) and BOP
(729 mg, 1.65 mmol) were combined in 5 mL DMF. After stirring overnight the reaction was concentrated, redissolved in EtOAc, washed with water, 5 % KHSO4, sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, EtOAc) provided 7-6 as a colorless waxy solid.
TLC Rf 0.39 (silica, EtOAc) lH-NMR (400 MHz, d6-DMSO). ~ 9.66 (,~, IH), ~s.l r (d, J = S Hz, IH), 1~ 7.76 (d, J = 9 Hz, lH), 7.6~ (s, lH), 7.25 (t, J = 7 Hz, 2H), 7.20-7.12 ~m, 3H), 6.88 (dd, J = 5, 1 Hz, lH), 4.01 (q, J = 7 Hz, 2H), 3.91 (AB d, J = 16 Hz, lH), 3.~S3 (AB d, J = 16 Hz, lH), 3.32 (s, 2H), 2.7~ (m, lH), 2.65-2.40 (m), l.g6-1.77 (m), 1.46 (s, 9H), 1.14 (t, J = 7 Hz, 3H), 0.77-0.70 (m, 4H).
2~) W ~97/26250 PCT~US97/00572 - ~7 :
HCI-H2N~ N~Jl~N~CO2Et N O
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-~-alanine ethyl ester hydrochloride (7-7) A solution of 7-6 (530 mg, 0.91 mmol) irl 4.6 rnL CH2C12 5 wa.s cooled to 0~C, 4.6 mL TFA was added, and after 1 h the reaction was warrned to RT for 90 min. After concentration and azeotroping with toluene the residue was purified by flash chromatography (silica, 10:1, EtOAc:NH3-saturated EtOH). The re~idue was dissolved in EtOAc, treated with I N HCI in ether, concentrated, then Iyophilized 10 from aq. acetonitrile, providing 7-7 as a gla~.sy solid.
TLC ]~f 0.25 (10:1, EtOAc:NH3-saturated EtOH) 1H-NMR (400 MHz, d6-DMSO): ~ 7.94 (br s, lH), 7.~6-7.~2 (m, 2H), 7.25 (t, J = 7 Hz, 2H), 7.20-7.13 (m, 3H), 6.g0-6.75 (m, 2H), 4.05 (m), 4.02 (~1, J = 7 Hz, 2H), 3.93 (AB d, J = 16 Hz, lH), 3.85 (AB d, J = 16 15 Hz, l]H), 2.78 (~In, lH), 2.65 (t, J = 7 Hz, 2H), 2.59 (t, J = 7 Hz, 2H), 2.55- .40 (m)~ 1.82 ((ln, J = 7 Hz, 2H), 1.80-1.70 (m, 2H), l.lS (t, J = 7 Hz, 3]H), 0.80-0.70 (m, 4H).
,.
W O 97/26250 PCTrUS97/00572 - 8~S -~ o H
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-~3-alanine (7-~) Ester 7-7 (100 mg, 0.1~ mmol) was di~solved in 4 mL
5 THF, then treated with 1 N LiOH (0.9 mL, 0.9 mmol). After stirring overnight the mixture was concentrated and purified by flash chromatography (silica, 15:20:1:1 EtOAc/EtOH/H2O/NH4OH) to provide 7-~ a~ a white solid.
TLC Rf 0.36 (silica, 15:20:1:1 EtOAc/EtOH/H2O/NH4OH) IH-NMR (400 MHz, d6-DMSO): ~ 7.gl (d, J = 9 Hz, IH), 7.77 (d, J = 5 Hz, lH), 7.27 (t, J = 7 Hz, 2H), 7.20-7.12 (m, 3H), 6.34 (dd, J = 5, 1 Hz, IH), 6.2~ (s, lH), 5.76 (br s, 2H), 4.03 (m, lH), 3.91 (AB d, J = 16 Hz, lH), 3.~s5 (AB d, J - 16 Hz, lH), 2.76 (m, lH), 2.65-2.50 (m), 2.45 (t, J = 7 Hz, 2H), 2.37 (d, J = 7 Hz, 2H), 1.82-1.60 (m), 0.77-0.6P~ (m, 1 5 4H).
W O 97/26250 PCT~US97/00572 ~=~
~7 H-BocNH~f~ ~N~CO2H H2N C4042Et N~ O
BOP, NMM
DMF
~ H-BocNH ~ N~J~ N ~CO2Et 1 N NaOH
EtOH
W O 97/26250 PCTrUS97/00572 ~ H
13ocNH~/~ ~ O H
TFA
Anisole ~H2CI2 H2N ~7 W O 97/26250 PCT~US97/OOS72 BocNH ~ ~ H -N O
4-(2-]30c-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-~2-(indol-3-yl)ethyll-,B-alanine ethyl e~ter (8-1) Acid 7-4 (lgO mg, 0.4~ mmol), amine 4-4 (130 mg, 0.50 mmol), NMM (1~3 ~L, 1.7 mmol) and BOP (253 mg, 0.57 mmol) were combined in 5 mL DMF. After stirring overnight the reaction wa.s concentrated, redissolved in EtOAc, washed with water ~3x), 10 %
KHS(~4, sat. NaHC03, and brine, dried (MgS04), filtered and concentrated. Flash chromatography (~ilica, gO % EtOAc/hexane) 10 provided g- 1 as a glas~;y solid.
TLC Rf 0.34 (silica, E~tOAc) lH-NMR (400 MHz, CDC13): ~ ~.10-~.00 (m, 2H), 7.79 (~, lH), 7.56 (d, J - 8 Hz, lH), 7.35-7.30 (m, 2H), 7.16 (t, J = ~ Hz, lH), 7.08 (t, J =
8 Hz, lH), 7.04 (s, lH), 6.gO (d, J = 5 Hz, lH), 6.71 (d, J - 9 Hz, lH), 4.29 (m, lH), 4.09 (q, J = 7 Hz, 2H), 3.99 (~, 2H), 2.85-2.70 (m, 4H), 2.66 (t, J = 7 Hz, 2H), 2.61 (t, J = 7 Hz, 2H), 2.51 (m), 2.05-1.87 (m, 4H), 1.53 (s, 9H), 1.21 (t, J = 7 Hz, 3H), 0.90-0.75 (m, 4H).
W O 97/26250 PCTrUS97/00572 ~1 BocNH ~ ~ ~,CO2H
4-(2-Boc-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-f2-(indol-3-yl)ethyll-~3-alanine (~s-2) Ester ~5-1 (223 mg, 0.36 mmol) wa~ dissolved in 4 mL
EtOH, then l N NaOH (0.90 mL, 0.90 mmol) was added. After a few hours the reaction was diluted with EtOAc, extracted with water and the pH of the aq. phase was adjusted to I with lO % KH$04. The a4ueous layer was extracted with EtOAc (2x), the combined organic layers were washed with brine, dried (MgS04), filtered and concentrated, providing 8-2asanoil.
TLC Rf 0.64 (silica, 9:1:1 CH2C12/MeOH/HOAc) /~ ~
H2N~ H~
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-[2-(indol-3-yl~ethyll-,B-alanine (8-3) Acid ~-2 (144 mg, 0.24 mmol) wa,s dissolved in 3 mL
CH2C12, then anisole (120 11L, 0.96 mmol) and TFA (3 mL) were added. After ca 1 h the reaction was concentrated. Flash chrornatography (silica, I 8: 10:1: 1 EtOAc/EtOH/H2O/NH40H, twice) - provided X-3 a~ a white ~iolid.
TLC Rf 0.29 (.~ilica, 1~:10:1:1 EtOAc/EtOH/H2O/NH4OH) lH-NMR (400 MHz, D20): ~ 7.~s8 (m, IH), 7.70 (m, lH), 7.53 (m, lH), 5 7.30-7.10 (m, 3H), 6.69 (m, lH), 6.58~(m, lH), 4.23 (m, IH), 3.99 (m, 2H), 2.~4 (m, 3H), 2.70 (m, 2H), 2.62 (m, 2H), 2.44 (m), 2.10-1.82 (m), 0.88-0.72 (m, 4H).
W 097126250 PCTrUS97/00572 ~7 H~, BocNH ~N~,CO2H HCI-H2N CO2Et N~ ~ / 9-1 BOP, NMM
DMF
~7 ~ H~, BocNH~ ~N~J~N CO2Et N O ~
TFA. CH2C12 0~C to RT
NH
LiOH
THF
H2N~ ~N~ C~2H
_ 95 _ BocNtl~ HCH3 4-(2-lBoc-amino-pyridin-4-yl)butanoyl -N-cyclopropylglycyl-3 (R)-methyl-,B-alanine ethyl ester (9-2) Acid 7-4 (100 mg, 0.26 mmol), amine hydrochloride 9-1 (see IJnited States Patent 5,2~1,5g5) (49 mg, 0.29 mmol), NMM (117 ,uL, 1.1 mmol) and BOP (176 mg, 0.40 mrnol) were combined in 1.3 mL DMF. After 3 d the mixture was concentrated, redissolved in EtOAc, washed with water (2x), 5 % KHSO4, sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography 10 (silica, EtOAc) provided 9-2 as a colorless oil.
TLC Rf 0.27 (silica, EtOAc) lH-NMR (400 MHz, CDC13): o 8.10 (d, J = 5 Hz, IH), ~.00 (br s, lH), 7.g5 ~.s, IH), 6.90 (dd,J = 5, 1 Hz, lH), 6.63 (d, J = ~ Hz, lH), 4.30 (m, lH), ~.12 ~q, J = 7 Hz, 2H~, 4.02 (AB d, J = 15 Hz, lH), 3.93 (AB d, J =
15 ~Iz, lH), 2.77 (m, lH), 2.73 (t, J = 7 Hz, 2H), 2.62 (t, J = 7 Hz, 2H), 2.47 (m, 2H), 2.04 (m, 2H), 1.53 (s, 9H), 1.25 (t, J=7 Hz, 3H), 1.20 (d, J = 7 Hz, 3H), 0.~X-0.7~ (m, 4H).
y ~ H~,, H2N~ H
BocN H N~ ~N ~ N ~CO2-t-Bu ~ o H H N-SO2~
4-(2-]3Oc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~3-alanirle t-butyl e~ster (5-Ps) Acid 5-5 (144 mg, 0.51 mmol), amine 5-7 (202 mg, 0.56 mmol), NMM (226 ,uL, 2.1 mmol) and BOP (241 mg, 0.77 mmol) were combined in 2.6 mL DMF. After stirring overnight the ~;olvent wa.s evaporated, the residue wa,s dissolved in EtOAc, washed with water, sat.
NaHCO3, 5 % KHSO4, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 10 % CHC13/EtOAc) provided S-g as an off-white glass.
TLC :Rf 0.22 (silica, 10 % CHC13/EtOAc) }H-NMR (400 MHz, d6-DMSO): ~ 9.53 (~, lH), 8.23 (br d, J = 7 Hz, lH), 7.92 (t, J = 6 Hz, lH), 7.91 (t, J = 6 Hz, lH), 7.76 (d, J = 7 Hz, 2H), 7.65-7.53 (m, 5H), 6.g7 (d, J = 7 Hz, lH), 3.~5 (br s, lH), 3.60 (t, J = 5 Hz, 2H), 3.20-3.10 (m, 2H), 2.60 (t, J = 7 Hz, 2H), 2.15 (t, J = 7 Hz, 2:H), l .85 (qn, J = 7 Hz, 2H), 1.46 (s, 9H), 1.1~ (s, 9H).
H2N N ' NJ~ N~CO2H
~'~ '~'~~ ~ H H H-S~2~
4-(2~ minopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-alanine (5-9) A solution of 5-8 (138 mg, 0.22 mrnol) in 1 mL CH2C12 was cooled to 0~C, treated with I mL TFA, and wa}med to RT for 5 h.
After concentration and azeotroping with toluene the residue was purified by fla.sh chromatography (.silica, 12:20: 1: 1, EtOAc/EtOH/H20/NH40H), providing 5-9 as a colorle~s glass.
W O 97/26250 PCT~US97/00572 TLC Rf 0.34 (silica,12:20: 1: 1, EtOAc/EtOH~H20~H40H) lH-NMR (400 MHz, D20): ~ 7.76 (dm, J = 7 Hz,2H),7.55-7.48 (m, 4H),6.69 (d, J - 7 Hz, lH),6.57 (d, J = ~ Hz, lH),3.72-3.62 (m,2H), 3.55 (dd, J = 8,5 Hz, lH),3.37 (dd, J = 13,8 Hz, lH),3.13 (dd, J = 13, 8 Hz, lH),2.63 (t, J = 7 Hz,2H),2.34 (t, J = 7 Hz,2H),1.96 ~qn, J = 7 Hz,2H).
W 09712625~ rCT~US97/00572 N~
1 0-5 H-Sar-OEt-HCI
BOP, NMM
DMF
Me "~ ~ N~,CO2Et N~ O
1 N NaOH ~,~
~tOH
Me H-,~ ~N~,CO2HH2N ~--C~2Et N~ O \
6-2 \/
BOP, NMM ~ H
DMF
Me ~ H-N~--U~ N ~CO2Et N O
-W O 97126250 PCTrUS97100572 - 7~S -/~\ H
/~ N
1 N NaOH
EtOH ~ J
Me ~ H-,~f ~N~JI' ~ CO2H
Me ~ ~ N~, CO2Et N~ O
4-(Pyridin-4-yl)butanoyl-sarcosine ethyl ester (6-1) 4-(4-Pyridyl)butanoic acid 10-5 (100 mg, 1.8 mmol), H-Sar-OEt-HCI (300 mg, 2.0 mmol), BOP (965 mg, 2.2 mmol~ and NMM
(700 IlL, 6.4 mmol) were combined in 9 mL DMF. After stirring 10 overnight the mixture wa.s diluted with EtOAc, washed with water (4x), sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated.
Flash chromatography (silica, 80% to 100 % Et~Ac/hexane) provided 6-1 as a cololless oil.
TLC Rf 0.44 (silica, 20 % MeOH/EtOAc) 1~ lH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major rotomer ~i 8.50 (d, J = 5 Hz, 2H), 7.14 (d, J = 5 Hz, 2H), 4.20 (~1, J = 7 Hz, 2H), 4.12 (s, 2H), 3.03 (s, 3H), 2.70 (t, J = 8 Hz, 2H), 2.39 (t, J = 7 Hz, 2H), 2.02 (~In, J = 7 Hz, 2H), 1.23 (t, J = 7 Hz, 3~I).
W O 97126250 PCTrUS97100~72 Me N~,CO2H
N ~ O
4-(Pyridin-4-yl)butanoyl-sarcosine (6-2) Ester 6-1 (324 mg, 1.22 mmol) wa~ dissolved in 6 mL
EtOH, then 1 N NaOH (2.4 mL, 2.4 mmol) wa.s added. After stirring overnight the mixture was concentrated, rediluted with EtOAc, extracted into 10 % KHSO4, then concentrated, providing acid 6-2, along with inorganic salts.
TLC l~f 0.16 (silica, 4: 1: 1 CH2C12/MeOH/HOAc) lH-NMR (400 MHz, CD30D): 1:1 mixture of amide rotomers, ~ g.41 10 (br s, 2H), 7.33 (m, 2H), 4.00/3.90 (s, 2~), 3.05/2.95 (s, 3H), 2.77-2.67 (m, 2]iI), 2.4~/2.37 (t, J = 7 Hz, 2H), 2.00-1.90 (m, 2H).
~=~
Me O H-~
N~ ~~ N'J~ N ~CO2Et 4-(Pyridin-4-yl)butanoyl-sarcosine-3 (R)-~2-(indol-3 -yl)ethyl]-,13-alanine 15 ethyl ester ~6-3) Acid 6-2 (2g~s mg, 1.22 mmol), amine 4-4 (318 mg, 1.22 mmol), BOP (647 mg, 1.5 mmol), and NMM (462 ~lL, 4.2 mrnol) were comb-ined in 6 mL DMF. After stirring overnight the mixture was diluted with EtOAc, washed with water (4x), sat. NaHCO3, and brine, 20 dried (MgSO4), filtered and concentrated. Flash chromatography (silica, EtOAc then 5 % MeOH/EtOAc) provided 6-3 as an orange oil.
TLC Rf 0.4 (20 % MeOH/EtOAc) W 097/26250 PCT~US97/OOS72 - ~0 -I H-NMR (400 MHz, CDC13): 4: 1 mixture of amide rotomers, major rotomer ~ 8.47 (br d, J = 5 H~, 2H), ~s.02 (br d, J = 6 Hz, 2H), 7.58 (dd, J = 16, g Hz, lH), 7.34 (dd, J = ~, 4 Hz, lH), 7.20-7.03 (m, 3H), 7.01 (s, IH), 6.72 (d, J = 9 Hz, lH), 4.33 (m, lH), 4.1 (t, J = 7 Hz, 3H), 3.9P, S (s, 2H), 3.05 (s, 3H), 2.90-2.45 (m), 2.39 (t, J = 7 Hz, 2H), 2.02-1.70 (m), 1.23 (t, J = 7 Hz, 3H)-/~\ H
/~
~ ~N--J~N~co2H
N O
4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-~2-(indol-3-yl)ethyl]-~-1 Q alanine (6-4) Ester 6-3 (400 mg, 0.~4 mmol) was dissolved in 4 mL
l~tOH, then 1 N NaOH (1.7 mL, 1.7 mmol) was added. After 90 min the reaction was neutralized with 1 N HCI (1.7 mL, 1.7 mmol) and concentrated to ~n oil. Flash chromatography (silica, 50: 1: 1 EtOH/H20/NH40H, then again with 12:10:1:1 EtOAc/EtOH/H20/NH40H) provided 6-4.
TLC Rf 0.17 (silica, 12:10:1:1 EtOAc/EtOH/H20/NE~40H) H-NMR (400 MH~, CD30D): 2:1 mixture of amide rotomers, ~ 8.3~S-~.2~ (m, 2H), 7.54-7.4~¢ (m, lH), 7.30-7.25 (m, 2H), 7.21-7.19 (m, lH), 2Q 7.07-6.92 (m, 3H), 4.36-4.27 (m, lH), 4.03-3.9~ (m, 2H), 3.06/2.93 (s, 3H), 2.~s6-2.60 (m, 4H), 2.52-2.32 (m), 2.05-l.g5 (m).
W O 97/26250 PCT~US97/OOS72 y Ethyl bromoacetate, TEA
CH2CI2, 0~C to RT
BocN H ~ C02H ~7 HN ~CO2Et BOP
NMM
DMF
BocNH~ ,N~,CO2Et N~ O
W O 97/262~0 PCT~US97/00572 SCHEME 7 (CONT'D) 1 N NaOH
MeOH
I~q BocNH ~N~,CO2H HCI-H2N ~CO2Et :Z~ \/ 7-S
BOP
NMM
DMF
~! ~
BocNH H~ CO2Et TFA, CH2CI2 W 097126250 PCTrUS97/00572 - ~3 -SCHEME 7 ~CONT'D) ,.
.
HCI-H2N ~ N~J~ N ~CO2Et LiOH, THF ¦
~7 ~ H -H2N~ N~J~ N~co2H
N ~7-8 HN ~,CO2Et N-Cyclopropyl~lvcine ethyl ester (7-2) Cyclopropylamine (12.1 mL, 175 mmol) and TEA (42 mL, 3~5 mmol) were combined at 0~C in 3~0 mL CH2C12, then ethyl 10 bromoacetate (19.4 mL, 175 mmol) was added dropwise. The reaction ~- wa~, warmed to RT for 3 h, then diluted with additional CH2C12, washed with water, .sat. NaHC03, and brine, dried (Na2S04), filtered and concentrated. Flash filtration (silica, 30 % EtOActhexane) provided 7-2 as a light yellow oil.
W O 97/26250 PCTnJS97/00572 - ~4 -TLC Rf 0.70 (~ilica, EtOAc) lH-NMR (400 MHz, CDC13): ~ 4.20 (q, J = 7 Hz, 2H), 3.45 (s, 2H), 2.23 (tt7 J = 6, 3 Hz, lH), 1.29 (t, J = 7 Hz, 3H), 0.43 (m, 2H), 0.36 (m, 2H).
y BocNH ~ N~CO2Et N~ O
4-(2-Boc-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycine ethyl ester (7-3) Acid 4 1 (0.86 g, 3.1 mmol), amine 7-2 (0.4~s g, 3.4 10 mmol), NMM (1.35 mL, 12.3 mrnol) and BOP (2.04 g, 4.61 mmol) were combined in 15 mL DMF. After stirring ovemight the mixture was concent~ated, redissolved in EtOAc, washed with water, 5 %
KHSO4, sat. NaHCO3 and brine, dried over MgSO4, filtered and concentrated. Flash chromatography (silica, 50 % EtC~Ac/hexane) 1~ provided 7-3 as a colorless oil.
TLC Rf 0.29 (silica, 50 % EtOAc~exane) lH-NMR (400 MHz, CDC13): ~ X.14 (d, J = S Hz, lH), 7.81 (.s, lH), 7.77 (br s, lH), 6.~4 (dd, J = 5, 1 Hz, lH), 4.18 (4, J = 7 Hz, 2H), 4.08 (s, 2H), 2.80 (tt, J = 7, 4 Hz, lH), 2.69 (t, J = 7 Hz, 2H), 2.60 (t, J=7 Hz, 20 2H)~ 2.02 (qn, J = 7 Hz, 2H), 1.53 (s, 9H), 1.27 (t, J = 7 Hz, 3H), 0.g3 (m, 2H), 0.72 (m, 2H).
W O 97126250 PCT~US97/00572 BocNH~ N~CO2H
N ~ O
4-(2-E~oc-amino-pyridin-4-yl~butanoyl-N-cyclopropyl~lycine (7-4) S Ester 7-3 (1.07 g, 2.64 mmol) wa~; dissolved in 26 mL
MeO~I, then treated with 1 N NaOH (6.6 m~, 6.6 rnrnol). After stirring overnight the reaction was concentrated, redissolved in water, the pH
was adjusted to 1 with 10 % KHSO4, then extracted with EtOAc (5x).
The a(~ueous pha.se was adjusted to pH 3 with a(l. NaOH, then reextr~cted with EtOAc (2x). The combined organic phases were washed with brine, dried (MgSO4), filtered and concentrated, providing 7-4 as a white foam.
TLC Rf 0.24 (silica, 19:1:1, CH2C12/MeOH/HOAc) lH-NMR (300 MHz, CDC13): ~ 9.15 (~r s, lH), 7.97 (d, J = 5 Hz, lH), 7.94 (:s, lH), 6.89 (dd, J = 5, 1 Hz, lH), 4.14 (,~, 2H), 2.81 (tt, J = 7, 3 Hz, lH),2.73(t,J=7Hz,2H),2.61 (t,J=7Hz,2H),2.04(qn,J=7 Hz, 2H), 1.51 (s, 9H), 0.85 (m, 2H), 0.76 (m, 2H).
W O 97i26250 PCTrUS97/00572 BocN H ~ N J~ N ~,CO2Et N O
4-(2-Boc-~mino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-13-alanine ethyl ester (7-6) Acid 7-4 (415 mg, 1.1 mmol), amine hydrochloride 7-5 (see procedure in EP 478 362 utilizing Boc-Gly(OEt) as starting material) (2~4 mg, 1.1 mmol), NMM (0.48 mL, 4.4 mmol) and BOP
(729 mg, 1.65 mmol) were combined in 5 mL DMF. After stirring overnight the reaction was concentrated, redissolved in EtOAc, washed with water, 5 % KHSO4, sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, EtOAc) provided 7-6 as a colorless waxy solid.
TLC Rf 0.39 (silica, EtOAc) lH-NMR (400 MHz, d6-DMSO). ~ 9.66 (,~, IH), ~s.l r (d, J = S Hz, IH), 1~ 7.76 (d, J = 9 Hz, lH), 7.6~ (s, lH), 7.25 (t, J = 7 Hz, 2H), 7.20-7.12 ~m, 3H), 6.88 (dd, J = 5, 1 Hz, lH), 4.01 (q, J = 7 Hz, 2H), 3.91 (AB d, J = 16 Hz, lH), 3.~S3 (AB d, J = 16 Hz, lH), 3.32 (s, 2H), 2.7~ (m, lH), 2.65-2.40 (m), l.g6-1.77 (m), 1.46 (s, 9H), 1.14 (t, J = 7 Hz, 3H), 0.77-0.70 (m, 4H).
2~) W ~97/26250 PCT~US97/00572 - ~7 :
HCI-H2N~ N~Jl~N~CO2Et N O
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-~-alanine ethyl ester hydrochloride (7-7) A solution of 7-6 (530 mg, 0.91 mmol) irl 4.6 rnL CH2C12 5 wa.s cooled to 0~C, 4.6 mL TFA was added, and after 1 h the reaction was warrned to RT for 90 min. After concentration and azeotroping with toluene the residue was purified by flash chromatography (silica, 10:1, EtOAc:NH3-saturated EtOH). The re~idue was dissolved in EtOAc, treated with I N HCI in ether, concentrated, then Iyophilized 10 from aq. acetonitrile, providing 7-7 as a gla~.sy solid.
TLC ]~f 0.25 (10:1, EtOAc:NH3-saturated EtOH) 1H-NMR (400 MHz, d6-DMSO): ~ 7.94 (br s, lH), 7.~6-7.~2 (m, 2H), 7.25 (t, J = 7 Hz, 2H), 7.20-7.13 (m, 3H), 6.g0-6.75 (m, 2H), 4.05 (m), 4.02 (~1, J = 7 Hz, 2H), 3.93 (AB d, J = 16 Hz, lH), 3.85 (AB d, J = 16 15 Hz, l]H), 2.78 (~In, lH), 2.65 (t, J = 7 Hz, 2H), 2.59 (t, J = 7 Hz, 2H), 2.55- .40 (m)~ 1.82 ((ln, J = 7 Hz, 2H), 1.80-1.70 (m, 2H), l.lS (t, J = 7 Hz, 3]H), 0.80-0.70 (m, 4H).
,.
W O 97/26250 PCTrUS97/00572 - 8~S -~ o H
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-~3-alanine (7-~) Ester 7-7 (100 mg, 0.1~ mmol) was di~solved in 4 mL
5 THF, then treated with 1 N LiOH (0.9 mL, 0.9 mmol). After stirring overnight the mixture was concentrated and purified by flash chromatography (silica, 15:20:1:1 EtOAc/EtOH/H2O/NH4OH) to provide 7-~ a~ a white solid.
TLC Rf 0.36 (silica, 15:20:1:1 EtOAc/EtOH/H2O/NH4OH) IH-NMR (400 MHz, d6-DMSO): ~ 7.gl (d, J = 9 Hz, IH), 7.77 (d, J = 5 Hz, lH), 7.27 (t, J = 7 Hz, 2H), 7.20-7.12 (m, 3H), 6.34 (dd, J = 5, 1 Hz, IH), 6.2~ (s, lH), 5.76 (br s, 2H), 4.03 (m, lH), 3.91 (AB d, J = 16 Hz, lH), 3.~s5 (AB d, J - 16 Hz, lH), 2.76 (m, lH), 2.65-2.50 (m), 2.45 (t, J = 7 Hz, 2H), 2.37 (d, J = 7 Hz, 2H), 1.82-1.60 (m), 0.77-0.6P~ (m, 1 5 4H).
W O 97/26250 PCT~US97/00572 ~=~
~7 H-BocNH~f~ ~N~CO2H H2N C4042Et N~ O
BOP, NMM
DMF
~ H-BocNH ~ N~J~ N ~CO2Et 1 N NaOH
EtOH
W O 97/26250 PCTrUS97/00572 ~ H
13ocNH~/~ ~ O H
TFA
Anisole ~H2CI2 H2N ~7 W O 97/26250 PCT~US97/OOS72 BocNH ~ ~ H -N O
4-(2-]30c-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-~2-(indol-3-yl)ethyll-,B-alanine ethyl e~ter (8-1) Acid 7-4 (lgO mg, 0.4~ mmol), amine 4-4 (130 mg, 0.50 mmol), NMM (1~3 ~L, 1.7 mmol) and BOP (253 mg, 0.57 mmol) were combined in 5 mL DMF. After stirring overnight the reaction wa.s concentrated, redissolved in EtOAc, washed with water ~3x), 10 %
KHS(~4, sat. NaHC03, and brine, dried (MgS04), filtered and concentrated. Flash chromatography (~ilica, gO % EtOAc/hexane) 10 provided g- 1 as a glas~;y solid.
TLC Rf 0.34 (silica, E~tOAc) lH-NMR (400 MHz, CDC13): ~ ~.10-~.00 (m, 2H), 7.79 (~, lH), 7.56 (d, J - 8 Hz, lH), 7.35-7.30 (m, 2H), 7.16 (t, J = ~ Hz, lH), 7.08 (t, J =
8 Hz, lH), 7.04 (s, lH), 6.gO (d, J = 5 Hz, lH), 6.71 (d, J - 9 Hz, lH), 4.29 (m, lH), 4.09 (q, J = 7 Hz, 2H), 3.99 (~, 2H), 2.85-2.70 (m, 4H), 2.66 (t, J = 7 Hz, 2H), 2.61 (t, J = 7 Hz, 2H), 2.51 (m), 2.05-1.87 (m, 4H), 1.53 (s, 9H), 1.21 (t, J = 7 Hz, 3H), 0.90-0.75 (m, 4H).
W O 97/26250 PCTrUS97/00572 ~1 BocNH ~ ~ ~,CO2H
4-(2-Boc-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-f2-(indol-3-yl)ethyll-~3-alanine (~s-2) Ester ~5-1 (223 mg, 0.36 mmol) wa~ dissolved in 4 mL
EtOH, then l N NaOH (0.90 mL, 0.90 mmol) was added. After a few hours the reaction was diluted with EtOAc, extracted with water and the pH of the aq. phase was adjusted to I with lO % KH$04. The a4ueous layer was extracted with EtOAc (2x), the combined organic layers were washed with brine, dried (MgS04), filtered and concentrated, providing 8-2asanoil.
TLC Rf 0.64 (silica, 9:1:1 CH2C12/MeOH/HOAc) /~ ~
H2N~ H~
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-[2-(indol-3-yl~ethyll-,B-alanine (8-3) Acid ~-2 (144 mg, 0.24 mmol) wa,s dissolved in 3 mL
CH2C12, then anisole (120 11L, 0.96 mmol) and TFA (3 mL) were added. After ca 1 h the reaction was concentrated. Flash chrornatography (silica, I 8: 10:1: 1 EtOAc/EtOH/H2O/NH40H, twice) - provided X-3 a~ a white ~iolid.
TLC Rf 0.29 (.~ilica, 1~:10:1:1 EtOAc/EtOH/H2O/NH4OH) lH-NMR (400 MHz, D20): ~ 7.~s8 (m, IH), 7.70 (m, lH), 7.53 (m, lH), 5 7.30-7.10 (m, 3H), 6.69 (m, lH), 6.58~(m, lH), 4.23 (m, IH), 3.99 (m, 2H), 2.~4 (m, 3H), 2.70 (m, 2H), 2.62 (m, 2H), 2.44 (m), 2.10-1.82 (m), 0.88-0.72 (m, 4H).
W 097126250 PCTrUS97/00572 ~7 H~, BocNH ~N~,CO2H HCI-H2N CO2Et N~ ~ / 9-1 BOP, NMM
DMF
~7 ~ H~, BocNH~ ~N~J~N CO2Et N O ~
TFA. CH2C12 0~C to RT
NH
LiOH
THF
H2N~ ~N~ C~2H
_ 95 _ BocNtl~ HCH3 4-(2-lBoc-amino-pyridin-4-yl)butanoyl -N-cyclopropylglycyl-3 (R)-methyl-,B-alanine ethyl ester (9-2) Acid 7-4 (100 mg, 0.26 mmol), amine hydrochloride 9-1 (see IJnited States Patent 5,2~1,5g5) (49 mg, 0.29 mmol), NMM (117 ,uL, 1.1 mmol) and BOP (176 mg, 0.40 mrnol) were combined in 1.3 mL DMF. After 3 d the mixture was concentrated, redissolved in EtOAc, washed with water (2x), 5 % KHSO4, sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated. Flash chromatography 10 (silica, EtOAc) provided 9-2 as a colorless oil.
TLC Rf 0.27 (silica, EtOAc) lH-NMR (400 MHz, CDC13): o 8.10 (d, J = 5 Hz, IH), ~.00 (br s, lH), 7.g5 ~.s, IH), 6.90 (dd,J = 5, 1 Hz, lH), 6.63 (d, J = ~ Hz, lH), 4.30 (m, lH), ~.12 ~q, J = 7 Hz, 2H~, 4.02 (AB d, J = 15 Hz, lH), 3.93 (AB d, J =
15 ~Iz, lH), 2.77 (m, lH), 2.73 (t, J = 7 Hz, 2H), 2.62 (t, J = 7 Hz, 2H), 2.47 (m, 2H), 2.04 (m, 2H), 1.53 (s, 9H), 1.25 (t, J=7 Hz, 3H), 1.20 (d, J = 7 Hz, 3H), 0.~X-0.7~ (m, 4H).
y ~ H~,, H2N~ H
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~3-a}anine ethyl ester (9-3) A solution of 9-2 (84 mg, 0.17 mmol) in 1 mL CH2C12 at - 0~C was treated with 1 mL TFA. After 3 h the mixture was warmed to RT fc~r 1 h, then concentrated and azeotroped with toluene. Flash W O 97/2G250 PCT~US97/00572 chromatography (~ilica, lS % NH3 satd. i-PrOH/EtOAc) and Iyophilization from aq. acetonitrile provided 9-3 as a semi-solid.
TLC Rf 0.19 (silica, lS % NH3 ~atd. i-PrOH/EtOAc) 1H-NMR (400 MHz, d6-DMSO): ~ 7.78 (d, J = 5 Hz, lH), 7.76 (d, J = 8 Hz, lH), 6.37 (dd, J = 5, 1 Hz, lH), 6.30 (.~, lH), 5.90 (br s, lH), 4.09 (m, J = 7 Hz, lH), 4.04 (4, J = 7 Hz, 2H), 3.~6 (AB d, J = 16 Hz, lH), 3.79 (AB d, J = 16 Hz, lH), 2.73 (m, lH), 2.45 (t, J = 7 Hz, 2H), 2.35 (ABX dd, J = 15, 7 Hz, lH), 1.77 (qn, J - 7 Hz, 2H), 1.17 (t, J = 7 Hz, 3H), 1.07 (d, J = 7 Hz, 3H), 0.76-0.67 (m, 4H).
H2N N ~CO2H
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~-alanine (9-4) Ester 9-3 (44 mg, 0.11 mrnol) was di~olved in 1.1 mL
THF, then 1 N LiOH (0.2~s mL, 0.2~ mmol) was added. After stirring ovemight the reaction mixture was loaded directly onto a fla~h chromatography column (silica, eluting with 7:20:1:1 EtOAc/EtOH/H20/NH40H) providing 9-4 as a white solid.
TLC Rf 0.62 (silica, 7:20:1:1 EtOAc/EtOH/H2O/NH4OH) lH-NMR (300 MHz, D20): ~i 7.72 (d, J = 7 Hz, lH), 6.90-6.~0 (m, 2H), 4.16 (hex, J = 7 IIz, lH), 4.04 (s, 2H), 2.~6 (tt, J = 7, 4 Hz, lH), 2.~S0-2.65 ~m, 4H), 2.41 (ABX dd, J = 14, 6 Hz, lH), 2.31 (ABX dd, J = 14, 7 Hz, lH), 1.9~ (qn, J = 7 Hz, 2H), 1.16 (d, J = 7 Hz, 3H), 0.~s9 (m, 2H), O.~sO (m, 2H).
W O 97126250 PCTrUS97/00572 CO2CH3 1 ) Na, MeOH CO2CH3 CO2CH3 N~ 50~C ~/~c02CH3 NaCI, H2O
TLC Rf 0.19 (silica, lS % NH3 ~atd. i-PrOH/EtOAc) 1H-NMR (400 MHz, d6-DMSO): ~ 7.78 (d, J = 5 Hz, lH), 7.76 (d, J = 8 Hz, lH), 6.37 (dd, J = 5, 1 Hz, lH), 6.30 (.~, lH), 5.90 (br s, lH), 4.09 (m, J = 7 Hz, lH), 4.04 (4, J = 7 Hz, 2H), 3.~6 (AB d, J = 16 Hz, lH), 3.79 (AB d, J = 16 Hz, lH), 2.73 (m, lH), 2.45 (t, J = 7 Hz, 2H), 2.35 (ABX dd, J = 15, 7 Hz, lH), 1.77 (qn, J - 7 Hz, 2H), 1.17 (t, J = 7 Hz, 3H), 1.07 (d, J = 7 Hz, 3H), 0.76-0.67 (m, 4H).
H2N N ~CO2H
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~-alanine (9-4) Ester 9-3 (44 mg, 0.11 mrnol) was di~olved in 1.1 mL
THF, then 1 N LiOH (0.2~s mL, 0.2~ mmol) was added. After stirring ovemight the reaction mixture was loaded directly onto a fla~h chromatography column (silica, eluting with 7:20:1:1 EtOAc/EtOH/H20/NH40H) providing 9-4 as a white solid.
TLC Rf 0.62 (silica, 7:20:1:1 EtOAc/EtOH/H2O/NH4OH) lH-NMR (300 MHz, D20): ~i 7.72 (d, J = 7 Hz, lH), 6.90-6.~0 (m, 2H), 4.16 (hex, J = 7 IIz, lH), 4.04 (s, 2H), 2.~6 (tt, J = 7, 4 Hz, lH), 2.~S0-2.65 ~m, 4H), 2.41 (ABX dd, J = 14, 6 Hz, lH), 2.31 (ABX dd, J = 14, 7 Hz, lH), 1.9~ (qn, J = 7 Hz, 2H), 1.16 (d, J = 7 Hz, 3H), 0.~s9 (m, 2H), O.~sO (m, 2H).
W O 97126250 PCTrUS97/00572 CO2CH3 1 ) Na, MeOH CO2CH3 CO2CH3 N~ 50~C ~/~c02CH3 NaCI, H2O
10-2 DMF, 170~C
N Q ~ s . 3 ~V~co2CH3 ~ 1~1 HC~ H2N~f~CH3 Ph~Br .10-6 60~C HN ~OCH3 10-5 + 10-8CH3CN ,Q \J~OCH3 W O 97/2625U PCTrUS97/00572 SCHEME 10 (~ONl INUED
1) 1N NaOH
2) HCI
H CH3CN ~ N~J~'OH
HCI H2N~--CO2Et N O
,~N~J~ NH~CO2Et N O
_ 99 _ 1) 1N NaOH
~ 2) HCI
N~ J~ NH ~CO2H
co2CH3 Methyl 2-(methoxycarbonyl)-4-(pyridin-4-yl)butyrate (10-3) To a stirred solution of elemental sodium (20 g, 840 mmol) and CH30H (600 ml) wa.~ added dimethyl malonate 10-l (135 ml, 1120 mmol). After 5 minutes, 4-vinyl pyridine 10-2 (15.3 ml, 140 mmoles) was added and the solution was heated to 50~C for 18 h. The reaction was cliluted with LtOAc and then washed with sat NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatoraphy (silica, 60%
~tOAc/hexanes) furnished the diether 10-3 (19.1 g) as a yellow oil.
TLC Rf = 0.43 (silica, EtOAc) lH NMR (400 MHz, CDC13) ~ 8.52 (d, J = 6 Hz, 2H), 7.12 (d, J = 6 Hz, 2H), 3.75 (s, 6H), 3.38 (t, J = ~ Hz, lH), 2.64 (t, J = ~s Hz, 2H), 2.24 (m, 2H).
W O 97/26250 PCTrUS97/00572 N~ CO2CH3 Methyl 4-(Pyridin-4-yl)butyrate (10-4) A solution of diester 10-3 (19.0 g, ~0.1 mrnol), H20 (1.45 S ml, 80.1 mmol), NaCI (10.5 g, 160.2 mmol) and DMF was heated to 170~C ~or 1~ h. The reaction was diluted with EtOAc and then washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Fla~sh chromatography (~ilica, 60% EtOAc/hexanes) afforded ester 10-4 as a brown oil.
10 TLC Rf 0.32 (silica, EtOAc) lH NMR (400 MHz, CD30D) ~ 8.40 (d, J = 6 Hz, 2H), 7.28 (d, J = 6 Hz, 2H), 3.64 (~, 3H), 2.67 (t, J = 8 Hz, 2H), 2.36 (t, J = 8 Hz, 2H), 1.94 (m, 2H).
4-(Pyridin-4-yl)butanoic acid (10-5) A solution of ester 10-4 (10.0 g, 56 mmol), lN NaOH (84 ml, 84 mrnole) and CH30H (200 ml) was stirred at ambient temperature 20 for 1.0 h. Concentrated HCl (7.0 ml, 84 mmol) was added followed by concentration. The residue wa.s dissolved in CHC13, dried (MgSO4) and concentrated to give acid 10-5 as a yellow solid.
TLCRf0.41 ~silica 10:1:1 CH2C12/MeOH/AcOH) lH NMR (400 MHz, CD30D) o 8.40 (d, J = 6 Hz, 2H), 7.30 (d, J = 6 25Hz, 2H), 2.71 (t, J = 8 Hz, 2H), 2.32 (t, J = 7 Hz, 2H), 1.93 (m, 2H).
W O 971262~0 PCTrUS97/00572 ,.
[~
N-(2-Phenethvl)~lycine methyl ester (10-8~
A solution of amine methyl ester 10-6 (1.0 g7 7.96 mmol), bromide 10-7 (1.09 ml7 7.96 mmole), NEt3 (3.33 ml2 23.9 mmol) and DMSO (25 ml) was heated to 60~C for 16 h. The reaction mixture was diluted with EtOAc and then washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Pla,sh chromatography (silica, 80%
EtOAc/hexanes) furnished ester 10-~S a,s a yellow oil.
TLC ~f 0.29 (silica, EtOAc) lH NMR (400 MH~;, CDC13) ~ 7.29 (m, 2H), 7.22 (m, 3H), 3.71 (s, 3H), 3.43 (s, 2H), 2.89 (m, 2H), 2.82 (m, 2H).
N--J~ocH3 N O
4-(Pyridin-4-yl)butanovl-N-(2-phenethyl)~lycine methyl e~ter (10-9) To a stirred .~;olution of acid 10-5 (342 mg, 2.07 mmol), NMM (910 ,ul, ~S.28 mmol) and CH3CN (15 ml) was added BOP reagent (1.01 g, 2.28 mmol). After 30 minutes, amine 10-8 (400 mg, 2.07 mmol) was added and stirring continued for an additional 18 h. The W 097/26250 PCT~US97/00572 reaction was diluted with EtOAc and then washed with ~at. NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (~ilica, EtOAc) furni.shed e,ster 10-9 as a yellow oil.
TLC Rf = 0.23 (silica, EtOAc) IH NMR (CD30D) 8.39 (d, J = 8 Hz, 2H), 7.14-7.29 (m, 7H), 4.~4 (s, 2H), 3.70 (s, 3H), 3.58 (t, J = 7 Hz, 2H), 2.~2 (t, J = 7 Hz, 2H), 2.66 (t, J = 8 Hz, 0.~6 H), 2.55 (t, J = ~ Hz, 1.44 H), 2.2~ (t, J = 7 Hz, 0.56 H), 2.10 (t, J = 7Hz, 1.44 H).
~ N ~J~OH
N~ O
4-(Pyridin-4-yl)butanoyl-N-(2-phenethyl) lycyl (10-10) A solution of ester 10-9 (500 mg, 1.47 mmole), lN NaOH
(2 ml, 2 mrnol) and CH30H (5 ml) was stirred at ambient temperature 15 for 2.0 h. Concentrated HCl (167 }Il, 2.0 mmol) was added followed by concentration. The residue was dissolved in CHC13, dried (MgSO4) and concentrated to give acid 10-10 as a white solid.
lH NMR (400 MHz, CD30D) ~ ~.47 (d, J = S Hz, 2H), 7.44 (m, 2H), 7.25 (m, 5H), 4.02 (s, 1.44 H), 3.96 (s, 0.56 H), 3.58 (m, 2H), 2.~4 (m, 20 2H), 2.74 (t, J = 8 Hz, 0.56 H), 2.63 (t, J = 8 Hz, 1.44 H), 2.33 (t, 3 =
7Hz, 0.56 H), 2.14 (t, J = 7 Hz, 1.44 H), 1.94 (m, 0.56 H), 1.79 (m, 1.44 H).
W O 9712625~ PCT~US97/00572 ~ NH ~~ Z
N O
4-(Pyridin-4-yl~butanoyl-N-(2-phenylethyl)glycyl-3 (R)-(2-phenethyl)-~-alanine ethyl ester (10- 11) S A solution of acid 10-10 (160 mg, 0.4903 mmol) amine 7-5 (164 mg, 0.49 mmol), NMM (216 ,ul, 1.96 mmol), BOP reagent (239 mg, 0.539 mmol) and CH3CN (S ml) was stirred at ambient temperature for 1 ~s h. The reaction wa~ diluted with EtOAc and then wa.shed with sat. I~aHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, EtOAc) furnished ester 10-11 (220 mg) as a colorless oil.
TLC Rf = 0.21 (silica, l~tOAc) lH l~IMR (400 MHz, CD30D) ~ 8.37 (m, 2H), 7.24 (m, 12H), 4.23 (m, lH), 4.0G (m, 2H), 3.95 (m, 2H), 2.84 (m, 2H), 2.56 (m, 6H), 2.34 (t, J
= 7 Hz, 0.56H), 2.16 (t, J = 7 Hz, 1.44 H), 1.83 (m, 0.56 H), 1.~¢1 (m, 1.44 H), 1.91 (m, 3H).
W 097/26250 PCTrUS97/00572 ~ H ~ CO2H
N O
4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-,B-alanine (10-12) S ~ solution of ester 10-11 (200 mg, 0.377g mmole), lN
NaOH (0.5 ml, 0.5 mmol) and CH30H was stirred at ambient temperature for l.S h followed by concentratioll. The crude acid was dissolved in H20, acidified with conc. HCI, concentrated and then azeotroped with toluene. Flash chromatography (silica, 20:20:1:1 EtOAc/EtOH/NH40H/H20) fumished acid 10-12 (100 mg) as a white solid.
TLC Rf 0.1 ~ (20:20: 1: 1 EtOAc/EtOH/NH40H/H20) 1 H NMR (400 MHz, D20) ~ ~.47 (d, J = 6 Hz, 1.36 H), g.43 (d, J = 6 Hz, 0.64 H), 7.71 (d, J = 6 Hz, 0.64 H), 7.66 (d, J = 6 Hz, 1.36 H), 7.20 (m, 10 H), 4.07 (m, lH), 3.~1 (s, 1.36 H), 3.73 (d, J = 6 Hz, 0.64 H), 3.51 (bt, 1.36 H), 3.43 (m, 0.64 H), 2.73 (m, 3H), 2.60 (t, J = 7 Hz, 1.36 H), 2.53 (m, 3.64 H), 2.18 (t, J = 7 Hz, 0.64 H), 1.7~s (m, 5.36 H).
W O 97/26250 PCT~US97/00572 , SCHEME 1 1 Ph Br~CO2Et 1 1-2 ~J
NEt3. CH2C12 HN C02Et 1 t-3 BOC-N ~ OH
N O
BOP, NMM, CH3CN
1N NaOH BOC-N~ a N~CO2Et ;~ 11-4 H O
BOC-N ~ ~,N J~OH
N~ O
N Q ~ s . 3 ~V~co2CH3 ~ 1~1 HC~ H2N~f~CH3 Ph~Br .10-6 60~C HN ~OCH3 10-5 + 10-8CH3CN ,Q \J~OCH3 W O 97/2625U PCTrUS97/00572 SCHEME 10 (~ONl INUED
1) 1N NaOH
2) HCI
H CH3CN ~ N~J~'OH
HCI H2N~--CO2Et N O
,~N~J~ NH~CO2Et N O
_ 99 _ 1) 1N NaOH
~ 2) HCI
N~ J~ NH ~CO2H
co2CH3 Methyl 2-(methoxycarbonyl)-4-(pyridin-4-yl)butyrate (10-3) To a stirred solution of elemental sodium (20 g, 840 mmol) and CH30H (600 ml) wa.~ added dimethyl malonate 10-l (135 ml, 1120 mmol). After 5 minutes, 4-vinyl pyridine 10-2 (15.3 ml, 140 mmoles) was added and the solution was heated to 50~C for 18 h. The reaction was cliluted with LtOAc and then washed with sat NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatoraphy (silica, 60%
~tOAc/hexanes) furnished the diether 10-3 (19.1 g) as a yellow oil.
TLC Rf = 0.43 (silica, EtOAc) lH NMR (400 MHz, CDC13) ~ 8.52 (d, J = 6 Hz, 2H), 7.12 (d, J = 6 Hz, 2H), 3.75 (s, 6H), 3.38 (t, J = ~ Hz, lH), 2.64 (t, J = ~s Hz, 2H), 2.24 (m, 2H).
W O 97/26250 PCTrUS97/00572 N~ CO2CH3 Methyl 4-(Pyridin-4-yl)butyrate (10-4) A solution of diester 10-3 (19.0 g, ~0.1 mrnol), H20 (1.45 S ml, 80.1 mmol), NaCI (10.5 g, 160.2 mmol) and DMF was heated to 170~C ~or 1~ h. The reaction was diluted with EtOAc and then washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Fla~sh chromatography (~ilica, 60% EtOAc/hexanes) afforded ester 10-4 as a brown oil.
10 TLC Rf 0.32 (silica, EtOAc) lH NMR (400 MHz, CD30D) ~ 8.40 (d, J = 6 Hz, 2H), 7.28 (d, J = 6 Hz, 2H), 3.64 (~, 3H), 2.67 (t, J = 8 Hz, 2H), 2.36 (t, J = 8 Hz, 2H), 1.94 (m, 2H).
4-(Pyridin-4-yl)butanoic acid (10-5) A solution of ester 10-4 (10.0 g, 56 mmol), lN NaOH (84 ml, 84 mrnole) and CH30H (200 ml) was stirred at ambient temperature 20 for 1.0 h. Concentrated HCl (7.0 ml, 84 mmol) was added followed by concentration. The residue wa.s dissolved in CHC13, dried (MgSO4) and concentrated to give acid 10-5 as a yellow solid.
TLCRf0.41 ~silica 10:1:1 CH2C12/MeOH/AcOH) lH NMR (400 MHz, CD30D) o 8.40 (d, J = 6 Hz, 2H), 7.30 (d, J = 6 25Hz, 2H), 2.71 (t, J = 8 Hz, 2H), 2.32 (t, J = 7 Hz, 2H), 1.93 (m, 2H).
W O 971262~0 PCTrUS97/00572 ,.
[~
N-(2-Phenethvl)~lycine methyl ester (10-8~
A solution of amine methyl ester 10-6 (1.0 g7 7.96 mmol), bromide 10-7 (1.09 ml7 7.96 mmole), NEt3 (3.33 ml2 23.9 mmol) and DMSO (25 ml) was heated to 60~C for 16 h. The reaction mixture was diluted with EtOAc and then washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Pla,sh chromatography (silica, 80%
EtOAc/hexanes) furnished ester 10-~S a,s a yellow oil.
TLC ~f 0.29 (silica, EtOAc) lH NMR (400 MH~;, CDC13) ~ 7.29 (m, 2H), 7.22 (m, 3H), 3.71 (s, 3H), 3.43 (s, 2H), 2.89 (m, 2H), 2.82 (m, 2H).
N--J~ocH3 N O
4-(Pyridin-4-yl)butanovl-N-(2-phenethyl)~lycine methyl e~ter (10-9) To a stirred .~;olution of acid 10-5 (342 mg, 2.07 mmol), NMM (910 ,ul, ~S.28 mmol) and CH3CN (15 ml) was added BOP reagent (1.01 g, 2.28 mmol). After 30 minutes, amine 10-8 (400 mg, 2.07 mmol) was added and stirring continued for an additional 18 h. The W 097/26250 PCT~US97/00572 reaction was diluted with EtOAc and then washed with ~at. NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (~ilica, EtOAc) furni.shed e,ster 10-9 as a yellow oil.
TLC Rf = 0.23 (silica, EtOAc) IH NMR (CD30D) 8.39 (d, J = 8 Hz, 2H), 7.14-7.29 (m, 7H), 4.~4 (s, 2H), 3.70 (s, 3H), 3.58 (t, J = 7 Hz, 2H), 2.~2 (t, J = 7 Hz, 2H), 2.66 (t, J = 8 Hz, 0.~6 H), 2.55 (t, J = ~ Hz, 1.44 H), 2.2~ (t, J = 7 Hz, 0.56 H), 2.10 (t, J = 7Hz, 1.44 H).
~ N ~J~OH
N~ O
4-(Pyridin-4-yl)butanoyl-N-(2-phenethyl) lycyl (10-10) A solution of ester 10-9 (500 mg, 1.47 mmole), lN NaOH
(2 ml, 2 mrnol) and CH30H (5 ml) was stirred at ambient temperature 15 for 2.0 h. Concentrated HCl (167 }Il, 2.0 mmol) was added followed by concentration. The residue was dissolved in CHC13, dried (MgSO4) and concentrated to give acid 10-10 as a white solid.
lH NMR (400 MHz, CD30D) ~ ~.47 (d, J = S Hz, 2H), 7.44 (m, 2H), 7.25 (m, 5H), 4.02 (s, 1.44 H), 3.96 (s, 0.56 H), 3.58 (m, 2H), 2.~4 (m, 20 2H), 2.74 (t, J = 8 Hz, 0.56 H), 2.63 (t, J = 8 Hz, 1.44 H), 2.33 (t, 3 =
7Hz, 0.56 H), 2.14 (t, J = 7 Hz, 1.44 H), 1.94 (m, 0.56 H), 1.79 (m, 1.44 H).
W O 9712625~ PCT~US97/00572 ~ NH ~~ Z
N O
4-(Pyridin-4-yl~butanoyl-N-(2-phenylethyl)glycyl-3 (R)-(2-phenethyl)-~-alanine ethyl ester (10- 11) S A solution of acid 10-10 (160 mg, 0.4903 mmol) amine 7-5 (164 mg, 0.49 mmol), NMM (216 ,ul, 1.96 mmol), BOP reagent (239 mg, 0.539 mmol) and CH3CN (S ml) was stirred at ambient temperature for 1 ~s h. The reaction wa~ diluted with EtOAc and then wa.shed with sat. I~aHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, EtOAc) furnished ester 10-11 (220 mg) as a colorless oil.
TLC Rf = 0.21 (silica, l~tOAc) lH l~IMR (400 MHz, CD30D) ~ 8.37 (m, 2H), 7.24 (m, 12H), 4.23 (m, lH), 4.0G (m, 2H), 3.95 (m, 2H), 2.84 (m, 2H), 2.56 (m, 6H), 2.34 (t, J
= 7 Hz, 0.56H), 2.16 (t, J = 7 Hz, 1.44 H), 1.83 (m, 0.56 H), 1.~¢1 (m, 1.44 H), 1.91 (m, 3H).
W 097/26250 PCTrUS97/00572 ~ H ~ CO2H
N O
4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-,B-alanine (10-12) S ~ solution of ester 10-11 (200 mg, 0.377g mmole), lN
NaOH (0.5 ml, 0.5 mmol) and CH30H was stirred at ambient temperature for l.S h followed by concentratioll. The crude acid was dissolved in H20, acidified with conc. HCI, concentrated and then azeotroped with toluene. Flash chromatography (silica, 20:20:1:1 EtOAc/EtOH/NH40H/H20) fumished acid 10-12 (100 mg) as a white solid.
TLC Rf 0.1 ~ (20:20: 1: 1 EtOAc/EtOH/NH40H/H20) 1 H NMR (400 MHz, D20) ~ ~.47 (d, J = 6 Hz, 1.36 H), g.43 (d, J = 6 Hz, 0.64 H), 7.71 (d, J = 6 Hz, 0.64 H), 7.66 (d, J = 6 Hz, 1.36 H), 7.20 (m, 10 H), 4.07 (m, lH), 3.~1 (s, 1.36 H), 3.73 (d, J = 6 Hz, 0.64 H), 3.51 (bt, 1.36 H), 3.43 (m, 0.64 H), 2.73 (m, 3H), 2.60 (t, J = 7 Hz, 1.36 H), 2.53 (m, 3.64 H), 2.18 (t, J = 7 Hz, 0.64 H), 1.7~s (m, 5.36 H).
W O 97/26250 PCT~US97/00572 , SCHEME 1 1 Ph Br~CO2Et 1 1-2 ~J
NEt3. CH2C12 HN C02Et 1 t-3 BOC-N ~ OH
N O
BOP, NMM, CH3CN
1N NaOH BOC-N~ a N~CO2Et ;~ 11-4 H O
BOC-N ~ ~,N J~OH
N~ O
PCT~US97/00572 SCHEME l l CONTINUED
1/2 H2SO4 H2N~CO2CH2Ph BOP, NMM CH3CN
40~C ~
BO ~ ~ H CH3 1N NaOH
C-N,~ N ,IlNH~--co2cH2ph 1 1-7 ~
~ ~ H CH3 BOC-N~ ~ N~JlNH~CO2H
N O
H2N~ ~N~I' NH ~C02H
W 097126250 PCT~US97/00572 ,. ¢~
HN ~,C02Et N-(2 Phenethyl)~lvcine ethyl ester ( 11 -3) A solution of amine 11-1 {20.0 g, 16~ mmol), NEt3 (47 ml, 330 mmol) in CH2C12 at 0~C was treated with bromide 11 2 (22.4 ml, 182 rnmol) followed by the removal of the cooling bath. After 1.0 h, the solution was washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Fla,~h chromatography (silica, 50% ~tO~c/hexanes) affor~ed ester 11-3 as a yellow oil.
TLC Rf 0.25 (silica, 50% EtOAc/~exanes) lH NMR (400 MHz, CD30D) 7.25 (m, 5H), 4.15 (~1, J = 7 Hz, 2El)1 3.37 (s, 2H), 2.~1 (m, 4H), 1.23 (t, J - 7 ~z, 3H).
BOC-N ~ ~ N~,CO2Et N~ O
[4-(2 -Boc-aminopyridin-4-yl)butanoyl]-N-(2-phenethyl)glycine ethyl ester (1 1-4) A solution of acid 4-1 (1.5 g, 5.35 mmol), amine 11-3 (1.66 g, ~.03 mmol), BOP reagent (2.61 g, 5.~9 mmol), NMM (3.0 ml, 21.4 rnmol~ and CH3CN {30 ml) was stirred at ambient temperature for W 097/26250 PCTrUS97/00572 - 10~ -l ~S h. The solution w~s diluted with EtOAc and then washed with H2O, .sat. NaHCO3, 10% KHSO4, brine, dried (MgSO4) and concentrated.
Fla~h chromatography (silica, 50% EtOAc/hexanes 80%
EtOAc/hexanes) furnished ester 11-4 as a yellow solid.
5 TLC Rf 0.35 (silica, 50% EtOAc/he~anes) lH NMR (400 MHz, CDC13) ~ 8.12 (d, J = 5 Hz, lH), 7.77 (m, 2H), 7.21 (m, 4H), 7.10 (d, J = 7 Hz, lH), 6.79 (m, lH), 4.18 (q, J = 7 Hz, 2H), 4.02 (s, 2H), 3.5~ (m, 2H), 2.82 (m, 2H), 2.62 (t, J = 7 Hz, 0.64 H), 2.57 (t, J = 7 Hz, 1.36 H), 2.15 (m, 2H), 1.91 (m, 2H), 1.52 (~, 9H), 1.27 (m, 3H).
BOC-N ~ ,N~J~OH
N~ O
~4-(2-BOC-Aminopyridin-4-yl)butanoyll-N-(2-phenethyl)~lycine (1 1-5) A solution of ester 11 4 (1.~ g, 3.~4 mmol), lN NaOH (6 ml, 6 mnlol) and EtOH (10 ml) was stirred at ambient temperature for 30 minutes. The solution was acidified with 10% KHSO4 and then extracted with EtOAc. The EtOAc phase was wa~shed with brine, dried (MgSO4) and concentrated to fumish acid 11-5 as a yellow solid.
20 TLC Rf 0.~0 (silica, 20:1:1 CH2C12/MeOH/AcOH) lH NMR (400 MHz, CD30D) ~ 8.12 ~m, lH), 7.17-7.29 (m, 7H), 4.06 (m, 2H), 3.61 (t, J = 7 Hz, 2H), 2.85 (t, J = 7 Hz, 2H), 2.~1 (m, 0.64 H), 2.63 (t,J=~Hz, 1.36H),2.35(t,J=7Hz,0.64H),2.14(t,J=7Hz, 1.36 H), 1.79 (m, 2H), 1.57 (s, 9H).
W O 97126250 PCTrUS97/00572 ., H ~ H CH3 BOC-N ~ ,N~JlNH~ co2cH2Ph N~ O
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methvl-,13-alanine benzyl ester (11-7) A solution of acid 11-5 (400 mg, 0.91 mmol), amine 11 -6 (available from Celgene) (285 mg, 1.09 mmol), BOP reagent (440 mg, 0.99 J mmol), NMM (502 ,ul, 3.63 mmol) and CH3CN (20 ml) was ~tirred at ambient temperature for 18 h. The solution was diluted with E~tOAc and then washed with H20, sat. NaHCO3, 10% KHSO4, brine, dried (MgSO4) and then concentrated. Flash chromatography (silica?
80% EtOAc/hexanes) fumished benzyl ester 11-7 as a yellow oil.
TLC Rf 0.49 (silica, E~tOAc) lH NMR (400 MHz, CD3OD) o 8.06 (d, J = S Hz, lH), 7.7 (s, 0.32 H), 7.68 (~, 0.68 H), 7.09-7.36 (m, 10 H), 6.83 (m, lH), 5.16 (s, 1.36 H), 5.08 (s, 0.64 H), 4.29 (m, lH), 3.93 (m, 2H), 3.51 (t, J = 7 Hz, 2H), 2.79 (4, J - 7 Hz, 2H), 2.50-2.61 (m, 4H), 2.25 (t, J = 8 Hz, 0.64 H), 2.09 (t, J = 7 Hz, 1.36 H), 1.73-1.84 (m, 4H), 1.51 (s, 9H), 1.25 (d, J =
7 Hz, 3H).
W 097/26250 PCT~US97/00572 J ..
H ~ ~ H CH3 BOC-N ~ N~ N~ co2H
N~ O
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-~-alanine (1 1-8) A solution of benzyl ester 11-7 (380 mg, 0.597 mmol), lN
NaOH (1 ml, 1.0 mmol) and EtOH (5 ml) was stirred at ambient temperature for 1.0 h. The solution was acidified with 10% KHSO4 and then extracted with EtOAc. The EtOAc phase was washed with brine, dried (MgSO4) and concentrated to furnish acid 11-8 as a yellow oil.
lH NMR (400 MHz, CD30D) ~ 8.13 (m, lH), 7.15-7.35 (m, 7H), 4.24 (m, lH), 3.91 (m, 2H), 3.58 (m, 2H), 2.~1 (m, 2.64 H), 2.62 (t, J = 8 Hz, 1.36 H), 2.36 (t, J = 7 Hz, 0.64 H), 2.14 (t, J = 7 Hz, 1.36 H), 1.79 (m, 2H), 1.57 (.~, 9H), 1.19 (m, 3H).
~ H CH3 H2N~f~ N'Jl' NH ~C02H
N~ O
4-(2-Aminopyridin-4-yl)butanoyl -N -(2-phenethyl)g Iycyl -3 (R)-methy 1-~-alanine (1 1-9) A solution of acid 11-~ (320 mg, 0.59 mmol) in CH2cl2 (5 ml) was treated with TFA (5 ml). After 1.0 h, the solution was concentrated and then azeotroped with toluene. Flash chromatography (silic.l, 10:10:1:1 EtOAc/EtOH/NH40H/H20) furnished amine I 1-10 (210 mg) as a white solid.
TLC Rf = 0.2~ (silica, 5:5:.5:.5 EtOAc/EtOH/NH40EI/H20) lH NMR (400 MHz, CD30DD) ~ 7.71 (d, J = 6 Hz, lH), 7.15-7.32 (m, 5H), I5.62-6.69 (m, 2H), 4.25 (m, lH), 3.99 (m, 2H), 3.58 (t, J = 7 Hz, 2H), 2.PS1 (m, 2H), 2.61 (t, J = 7 Hz, 0.64 H), 2.31-2.51 (m, 3.36 H), 2.12 (td, J = 3 Hz, 7 Hz, 1.36 H), l.P~9 (t, J = ~ Hz, 0.64 H), 1.79 (m, 2H), 1.19 (m, 3H).
PCTrUS97/00572 SC~DEME 12 OH B ~OCH2CH3 N~, 12-1 0 r ~0~ \--~f QC H2CH3 N CS2CO3, DMF 12-2 O
KOH, THF, H20 ~o ~O K+
12-3 ~
EDC, HOBT,.11-~
DMF
Ph ~0/\~ OCH2CH3 12-4 ~
KOH, THF, H20 Ph ~o~~~~f ~O K+
W O 97)262'tO
SCHEME 12 ~ONTINUED
-Ph -~\,CO2CH2CH3 EDC, HOBT, DMF HCI~H2N 7 5 Ph ~h ~o ~ J ~ -~ CO~CH~CH~
KOH, H20, H+
Ph Ph CO H
~o~ - l~ocH2cH3 12-1 ~
W O 97/26250 PCT~US97/00572 Ethyl-4-(4-Pyridvloxy)butyrate (12-2) A mixture of 4-hydroxypyridine (10 g, lûS mmol), ethyl 4-bromobutyrate 12-1 (15.0 ml, 105 mmol) arld Cs2CO3 (34.2 g, 105 mmol) in DMF (100 ml) was stirred at room temperature for 24 h. The 5 reaction was filtered and the filtrate diluted with ethyl acetate (30Q ml) and washed with water (4 x 100 ml) and brine (100 ml) then dried (Na2 SO4), filtered, and evaporated. The resulting oil was purified by chromatography on silica gel (3% CH3OH/CH2C12) to give 12-2 as a colorless glass.
10 TLC Rf 0.45 (silica, 5% CH30H/CH2C12) lH NMR (300 MHz, CDC13) ~ P~.41 (d, J=6.~Hz, 2H), 6.~s3 (d, J=6Hz, 2H), 4.16 (q, J--7Hz, 2H), 4.07 (t, J=7Hz, 2~), 2.52 (t, J-7Hz, 2H), 2.~S1 (t, J=7Hz, 2H), 1.23 (t, J=7.0Hz, 3H).
N~
~o~C02K
.1 2-3 Potassium 4-(4-pyridyloxy)butyrate ( 12-3) 'rhe e,ster 12-1 (2.5 g, 12.0 mmol) was dissolved in 10 ml THF and treated with 0.5 N KOH (24 ml, 12.0 mmol) and H20 (10 ml).
20 The resulting solution was stirred at room temperature for 78 h then evaporated at reduced pressure to give 12 2 as a white solid.
lH NMR (300 MHz, D20) â 8.19 (d, J = 6.~ Hz, 2H), 6.~3 (d, J = 6.g Hz, 2H), 6.~3 (d, J = 6.~ Hz, 2H), 3.96 (t, J = 7.1 Hz, 2H), 2.1~ (t, J =
7.1 Hz, 2H), 1.93 (m, 2H).
W 097t2625~ PCT~US97/00572 - l l 5 -,.
Ph --0~ ~OCH2CH3 12-4 o 4-(4-Pyridyloxy)butyrate-N-(2-phenethyl)glycine el:hyl e.ster ~12-4) The alkoxy pyridine 12-3 (29~ mg, 1.36 mmol) and amine S 1 1-3 ~450 mg, 1.36 mmol) were combined with EDC (260 mg, 136 mmol), HOBT (20~ mg, 136 mmol), in DMF (30 ml) and stirred at room temperature for 16 h. The solution was then diluted with ethyl acetate (200 ml) and washed with sa~. NaHCO3 (2 x 100 ml) and brine (100 ml). The organic layer was dried (Na2S04), filtered, and evaporated and the residue chromatographed on silica gel (3~o CH3OH/CH2C12) to give 12-4 as a colorless glas.s.
lH NMR (300 MHz, CDC13) ~ 8.41 (d, J = 6.5 Hz, 2H), 7.25 (m, SH), 6.7~ (d, J = 6.5 Hz, 2H), 61.23 (m, 2H), 4.02 (s, 2H), 4.00 (m, 2H), 3.63 (m, 2H), 3.41 (m, 2H), 2.15 (m, 2H), 1.31 (m, 3H).
Ph ~0 f ~0 K+
4-(4-Pvridyloxy)butyrate-N-(2-phenethyl)glycine potassium salt (12-5) Ester 12-4 (360 mg, 0.97 mmol) was hydrolyzed in 0.5 N
KOH (1.94 ml, 0.97 mmol) to give the potassium salt 12-5 as a white ~solid.
lH NMR (300 MHz, DMSO-d6) ~ ~.3~ (d, J = 6.5 Hz, 2H), 7.25 (m, 5H), 6.93 (d, J = 6.5 Hz, 2H), 4.016 m, 2H), 3.45 (m, 2H), 3.25 (s~ 2H), 2.6~ (m, 2H), 2.21 (m, 2H), 1.~6 (m, 2H).
W O 97/26250 PCTrUS97/00572 Ph Ph ~O N~ N ~CO2CH2CH3 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanine ethyl e,ster (12-6) Acid salt 12-5 (352 mg, 0.93 mmol) and amino ester 7-S
(240 mg, 193 mmol), HOBT (142 mg, 0.93 mmol), EDC (19~ mg, 0.93 mmol), and triethylamine (130 ~11, 0.93 mmol) was dissol~ed in DMF
(15 ml) and stirred at room temperature for 18s h. The solution was diluted with ethyl acetate (200 ml) washed with sat. NaHCO3, water and brine (100 ml each), dried (Na2SO4) and concentrated to give a colorless oil. Chromatography on silica gel afforded 12-6 a.s a colorless gla~s.
TLC Rf 0.50 (silica, 3% CH3OH/CH2C12) lH NMR (300 MHz, CDC13) ~ 8.40 (d, J = 6.6 Hz, 2H), 7.25 (m, 10H), 6.85 (t, J = 7.4 Hz, lH), 7.25 (d, J = 6.6 Hz, 2H), 4.25 (m, lH), 4.1~
(m, 2H), 4.00 (m, 2H), 3.60 (m, 2H), 2.95 (m, 2H), 2.63 (m, 2H), 2.58s (m, 2H), 2.40 (m, 2H), 2.0P~ (m, 2H), 1.85 (m, 2H), 1.16 (m, 3H).
Ph Ph ~o N~J~ N~CO~H
4-(4-:~Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanille (12-7) Ester 12 6 (123 mg, 0.23 mmol) was hydrolyzed with 0.5 N KOH and the acid was isolated as its TFA salt following preparative reverse phase chromatography.
lH NMR (300 MHz, CD30D) ~ P~.63 (d, J = 6.5 Hz, 2H), 7.52 (d, J =
6.5 Hz, 2H), 7.20 (m, 10H), 4.41 (m, lH), 4.32 (m, 2H), 4.01 (m, 2H), 3.gl l(m, 2H), 2.~5 ~m, 2H), 2.63 (m, 2H), 2.30 ~m, 2H), 2.41 (m, 2H), 2.20 ~m, 2H), 1.~S5 (m, 2H).
W O 97/26250 PCTrUS97/00572 SCHE~ME 13 CH3 ~, CO2tBu 3 2 ~ HN ~CO2tBu HCI
NMP, NMM
HCI . N~ ,N~ CO2H HCI, EtOAc ~N~ CO2tBu .~CH3 CH3 13-2 CH3NH CO2Et HCI
NMM, EDC, HOBT, DMF
N~ O N~ O
CH3 CH3 O . CH3 CH3 O
1/2 H2SO4 H2N~CO2CH2Ph BOP, NMM CH3CN
40~C ~
BO ~ ~ H CH3 1N NaOH
C-N,~ N ,IlNH~--co2cH2ph 1 1-7 ~
~ ~ H CH3 BOC-N~ ~ N~JlNH~CO2H
N O
H2N~ ~N~I' NH ~C02H
W 097126250 PCT~US97/00572 ,. ¢~
HN ~,C02Et N-(2 Phenethyl)~lvcine ethyl ester ( 11 -3) A solution of amine 11-1 {20.0 g, 16~ mmol), NEt3 (47 ml, 330 mmol) in CH2C12 at 0~C was treated with bromide 11 2 (22.4 ml, 182 rnmol) followed by the removal of the cooling bath. After 1.0 h, the solution was washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Fla,~h chromatography (silica, 50% ~tO~c/hexanes) affor~ed ester 11-3 as a yellow oil.
TLC Rf 0.25 (silica, 50% EtOAc/~exanes) lH NMR (400 MHz, CD30D) 7.25 (m, 5H), 4.15 (~1, J = 7 Hz, 2El)1 3.37 (s, 2H), 2.~1 (m, 4H), 1.23 (t, J - 7 ~z, 3H).
BOC-N ~ ~ N~,CO2Et N~ O
[4-(2 -Boc-aminopyridin-4-yl)butanoyl]-N-(2-phenethyl)glycine ethyl ester (1 1-4) A solution of acid 4-1 (1.5 g, 5.35 mmol), amine 11-3 (1.66 g, ~.03 mmol), BOP reagent (2.61 g, 5.~9 mmol), NMM (3.0 ml, 21.4 rnmol~ and CH3CN {30 ml) was stirred at ambient temperature for W 097/26250 PCTrUS97/00572 - 10~ -l ~S h. The solution w~s diluted with EtOAc and then washed with H2O, .sat. NaHCO3, 10% KHSO4, brine, dried (MgSO4) and concentrated.
Fla~h chromatography (silica, 50% EtOAc/hexanes 80%
EtOAc/hexanes) furnished ester 11-4 as a yellow solid.
5 TLC Rf 0.35 (silica, 50% EtOAc/he~anes) lH NMR (400 MHz, CDC13) ~ 8.12 (d, J = 5 Hz, lH), 7.77 (m, 2H), 7.21 (m, 4H), 7.10 (d, J = 7 Hz, lH), 6.79 (m, lH), 4.18 (q, J = 7 Hz, 2H), 4.02 (s, 2H), 3.5~ (m, 2H), 2.82 (m, 2H), 2.62 (t, J = 7 Hz, 0.64 H), 2.57 (t, J = 7 Hz, 1.36 H), 2.15 (m, 2H), 1.91 (m, 2H), 1.52 (~, 9H), 1.27 (m, 3H).
BOC-N ~ ,N~J~OH
N~ O
~4-(2-BOC-Aminopyridin-4-yl)butanoyll-N-(2-phenethyl)~lycine (1 1-5) A solution of ester 11 4 (1.~ g, 3.~4 mmol), lN NaOH (6 ml, 6 mnlol) and EtOH (10 ml) was stirred at ambient temperature for 30 minutes. The solution was acidified with 10% KHSO4 and then extracted with EtOAc. The EtOAc phase was wa~shed with brine, dried (MgSO4) and concentrated to fumish acid 11-5 as a yellow solid.
20 TLC Rf 0.~0 (silica, 20:1:1 CH2C12/MeOH/AcOH) lH NMR (400 MHz, CD30D) ~ 8.12 ~m, lH), 7.17-7.29 (m, 7H), 4.06 (m, 2H), 3.61 (t, J = 7 Hz, 2H), 2.85 (t, J = 7 Hz, 2H), 2.~1 (m, 0.64 H), 2.63 (t,J=~Hz, 1.36H),2.35(t,J=7Hz,0.64H),2.14(t,J=7Hz, 1.36 H), 1.79 (m, 2H), 1.57 (s, 9H).
W O 97126250 PCTrUS97/00572 ., H ~ H CH3 BOC-N ~ ,N~JlNH~ co2cH2Ph N~ O
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methvl-,13-alanine benzyl ester (11-7) A solution of acid 11-5 (400 mg, 0.91 mmol), amine 11 -6 (available from Celgene) (285 mg, 1.09 mmol), BOP reagent (440 mg, 0.99 J mmol), NMM (502 ,ul, 3.63 mmol) and CH3CN (20 ml) was ~tirred at ambient temperature for 18 h. The solution was diluted with E~tOAc and then washed with H20, sat. NaHCO3, 10% KHSO4, brine, dried (MgSO4) and then concentrated. Flash chromatography (silica?
80% EtOAc/hexanes) fumished benzyl ester 11-7 as a yellow oil.
TLC Rf 0.49 (silica, E~tOAc) lH NMR (400 MHz, CD3OD) o 8.06 (d, J = S Hz, lH), 7.7 (s, 0.32 H), 7.68 (~, 0.68 H), 7.09-7.36 (m, 10 H), 6.83 (m, lH), 5.16 (s, 1.36 H), 5.08 (s, 0.64 H), 4.29 (m, lH), 3.93 (m, 2H), 3.51 (t, J = 7 Hz, 2H), 2.79 (4, J - 7 Hz, 2H), 2.50-2.61 (m, 4H), 2.25 (t, J = 8 Hz, 0.64 H), 2.09 (t, J = 7 Hz, 1.36 H), 1.73-1.84 (m, 4H), 1.51 (s, 9H), 1.25 (d, J =
7 Hz, 3H).
W 097/26250 PCT~US97/00572 J ..
H ~ ~ H CH3 BOC-N ~ N~ N~ co2H
N~ O
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-~-alanine (1 1-8) A solution of benzyl ester 11-7 (380 mg, 0.597 mmol), lN
NaOH (1 ml, 1.0 mmol) and EtOH (5 ml) was stirred at ambient temperature for 1.0 h. The solution was acidified with 10% KHSO4 and then extracted with EtOAc. The EtOAc phase was washed with brine, dried (MgSO4) and concentrated to furnish acid 11-8 as a yellow oil.
lH NMR (400 MHz, CD30D) ~ 8.13 (m, lH), 7.15-7.35 (m, 7H), 4.24 (m, lH), 3.91 (m, 2H), 3.58 (m, 2H), 2.~1 (m, 2.64 H), 2.62 (t, J = 8 Hz, 1.36 H), 2.36 (t, J = 7 Hz, 0.64 H), 2.14 (t, J = 7 Hz, 1.36 H), 1.79 (m, 2H), 1.57 (.~, 9H), 1.19 (m, 3H).
~ H CH3 H2N~f~ N'Jl' NH ~C02H
N~ O
4-(2-Aminopyridin-4-yl)butanoyl -N -(2-phenethyl)g Iycyl -3 (R)-methy 1-~-alanine (1 1-9) A solution of acid 11-~ (320 mg, 0.59 mmol) in CH2cl2 (5 ml) was treated with TFA (5 ml). After 1.0 h, the solution was concentrated and then azeotroped with toluene. Flash chromatography (silic.l, 10:10:1:1 EtOAc/EtOH/NH40H/H20) furnished amine I 1-10 (210 mg) as a white solid.
TLC Rf = 0.2~ (silica, 5:5:.5:.5 EtOAc/EtOH/NH40EI/H20) lH NMR (400 MHz, CD30DD) ~ 7.71 (d, J = 6 Hz, lH), 7.15-7.32 (m, 5H), I5.62-6.69 (m, 2H), 4.25 (m, lH), 3.99 (m, 2H), 3.58 (t, J = 7 Hz, 2H), 2.PS1 (m, 2H), 2.61 (t, J = 7 Hz, 0.64 H), 2.31-2.51 (m, 3.36 H), 2.12 (td, J = 3 Hz, 7 Hz, 1.36 H), l.P~9 (t, J = ~ Hz, 0.64 H), 1.79 (m, 2H), 1.19 (m, 3H).
PCTrUS97/00572 SC~DEME 12 OH B ~OCH2CH3 N~, 12-1 0 r ~0~ \--~f QC H2CH3 N CS2CO3, DMF 12-2 O
KOH, THF, H20 ~o ~O K+
12-3 ~
EDC, HOBT,.11-~
DMF
Ph ~0/\~ OCH2CH3 12-4 ~
KOH, THF, H20 Ph ~o~~~~f ~O K+
W O 97)262'tO
SCHEME 12 ~ONTINUED
-Ph -~\,CO2CH2CH3 EDC, HOBT, DMF HCI~H2N 7 5 Ph ~h ~o ~ J ~ -~ CO~CH~CH~
KOH, H20, H+
Ph Ph CO H
~o~ - l~ocH2cH3 12-1 ~
W O 97/26250 PCT~US97/00572 Ethyl-4-(4-Pyridvloxy)butyrate (12-2) A mixture of 4-hydroxypyridine (10 g, lûS mmol), ethyl 4-bromobutyrate 12-1 (15.0 ml, 105 mmol) arld Cs2CO3 (34.2 g, 105 mmol) in DMF (100 ml) was stirred at room temperature for 24 h. The 5 reaction was filtered and the filtrate diluted with ethyl acetate (30Q ml) and washed with water (4 x 100 ml) and brine (100 ml) then dried (Na2 SO4), filtered, and evaporated. The resulting oil was purified by chromatography on silica gel (3% CH3OH/CH2C12) to give 12-2 as a colorless glass.
10 TLC Rf 0.45 (silica, 5% CH30H/CH2C12) lH NMR (300 MHz, CDC13) ~ P~.41 (d, J=6.~Hz, 2H), 6.~s3 (d, J=6Hz, 2H), 4.16 (q, J--7Hz, 2H), 4.07 (t, J=7Hz, 2~), 2.52 (t, J-7Hz, 2H), 2.~S1 (t, J=7Hz, 2H), 1.23 (t, J=7.0Hz, 3H).
N~
~o~C02K
.1 2-3 Potassium 4-(4-pyridyloxy)butyrate ( 12-3) 'rhe e,ster 12-1 (2.5 g, 12.0 mmol) was dissolved in 10 ml THF and treated with 0.5 N KOH (24 ml, 12.0 mmol) and H20 (10 ml).
20 The resulting solution was stirred at room temperature for 78 h then evaporated at reduced pressure to give 12 2 as a white solid.
lH NMR (300 MHz, D20) â 8.19 (d, J = 6.~ Hz, 2H), 6.~3 (d, J = 6.g Hz, 2H), 6.~3 (d, J = 6.~ Hz, 2H), 3.96 (t, J = 7.1 Hz, 2H), 2.1~ (t, J =
7.1 Hz, 2H), 1.93 (m, 2H).
W 097t2625~ PCT~US97/00572 - l l 5 -,.
Ph --0~ ~OCH2CH3 12-4 o 4-(4-Pyridyloxy)butyrate-N-(2-phenethyl)glycine el:hyl e.ster ~12-4) The alkoxy pyridine 12-3 (29~ mg, 1.36 mmol) and amine S 1 1-3 ~450 mg, 1.36 mmol) were combined with EDC (260 mg, 136 mmol), HOBT (20~ mg, 136 mmol), in DMF (30 ml) and stirred at room temperature for 16 h. The solution was then diluted with ethyl acetate (200 ml) and washed with sa~. NaHCO3 (2 x 100 ml) and brine (100 ml). The organic layer was dried (Na2S04), filtered, and evaporated and the residue chromatographed on silica gel (3~o CH3OH/CH2C12) to give 12-4 as a colorless glas.s.
lH NMR (300 MHz, CDC13) ~ 8.41 (d, J = 6.5 Hz, 2H), 7.25 (m, SH), 6.7~ (d, J = 6.5 Hz, 2H), 61.23 (m, 2H), 4.02 (s, 2H), 4.00 (m, 2H), 3.63 (m, 2H), 3.41 (m, 2H), 2.15 (m, 2H), 1.31 (m, 3H).
Ph ~0 f ~0 K+
4-(4-Pvridyloxy)butyrate-N-(2-phenethyl)glycine potassium salt (12-5) Ester 12-4 (360 mg, 0.97 mmol) was hydrolyzed in 0.5 N
KOH (1.94 ml, 0.97 mmol) to give the potassium salt 12-5 as a white ~solid.
lH NMR (300 MHz, DMSO-d6) ~ ~.3~ (d, J = 6.5 Hz, 2H), 7.25 (m, 5H), 6.93 (d, J = 6.5 Hz, 2H), 4.016 m, 2H), 3.45 (m, 2H), 3.25 (s~ 2H), 2.6~ (m, 2H), 2.21 (m, 2H), 1.~6 (m, 2H).
W O 97/26250 PCTrUS97/00572 Ph Ph ~O N~ N ~CO2CH2CH3 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanine ethyl e,ster (12-6) Acid salt 12-5 (352 mg, 0.93 mmol) and amino ester 7-S
(240 mg, 193 mmol), HOBT (142 mg, 0.93 mmol), EDC (19~ mg, 0.93 mmol), and triethylamine (130 ~11, 0.93 mmol) was dissol~ed in DMF
(15 ml) and stirred at room temperature for 18s h. The solution was diluted with ethyl acetate (200 ml) washed with sat. NaHCO3, water and brine (100 ml each), dried (Na2SO4) and concentrated to give a colorless oil. Chromatography on silica gel afforded 12-6 a.s a colorless gla~s.
TLC Rf 0.50 (silica, 3% CH3OH/CH2C12) lH NMR (300 MHz, CDC13) ~ 8.40 (d, J = 6.6 Hz, 2H), 7.25 (m, 10H), 6.85 (t, J = 7.4 Hz, lH), 7.25 (d, J = 6.6 Hz, 2H), 4.25 (m, lH), 4.1~
(m, 2H), 4.00 (m, 2H), 3.60 (m, 2H), 2.95 (m, 2H), 2.63 (m, 2H), 2.58s (m, 2H), 2.40 (m, 2H), 2.0P~ (m, 2H), 1.85 (m, 2H), 1.16 (m, 3H).
Ph Ph ~o N~J~ N~CO~H
4-(4-:~Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-alanille (12-7) Ester 12 6 (123 mg, 0.23 mmol) was hydrolyzed with 0.5 N KOH and the acid was isolated as its TFA salt following preparative reverse phase chromatography.
lH NMR (300 MHz, CD30D) ~ P~.63 (d, J = 6.5 Hz, 2H), 7.52 (d, J =
6.5 Hz, 2H), 7.20 (m, 10H), 4.41 (m, lH), 4.32 (m, 2H), 4.01 (m, 2H), 3.gl l(m, 2H), 2.~5 ~m, 2H), 2.63 (m, 2H), 2.30 ~m, 2H), 2.41 (m, 2H), 2.20 ~m, 2H), 1.~S5 (m, 2H).
W O 97/26250 PCTrUS97/00572 SCHE~ME 13 CH3 ~, CO2tBu 3 2 ~ HN ~CO2tBu HCI
NMP, NMM
HCI . N~ ,N~ CO2H HCI, EtOAc ~N~ CO2tBu .~CH3 CH3 13-2 CH3NH CO2Et HCI
NMM, EDC, HOBT, DMF
N~ O N~ O
CH3 CH3 O . CH3 CH3 O
EDC, HOBT, DMF
j, - 1 1 9 -SCHEME 13 CONTINUEr) ~.
Ph Nl~'N, ~~N~H - CO2Et KOH
N~ O H - CO
HN--,CO2tBu tert-13utyl 3-N-methylaminopropionate ( 13-1 ) Tert-butyl acrylate (15 g, 1 17 mmol) was added to a solution or methanol saturated with CH3NH2 (300 ml) and stirred at 10 room temperature for 16 h. The solution was evaporated to afford 13-1 as a colorle~s liquid.
W 097/26250 PCT~US97/00572 H NMR (300 MHz, CDC13) ~ 2.81 (t, J = 7.2 Hz, 2H), 2.43 (t, J = 7.2 ~z, 2~), 1.45 (.s, 9~
N~N~ CO2tBu tert-Butyl 3-~(N-methyl-N-(4-pyridyl)laminopropionate (13-2) A mixture of 4-chloropyrid~ne hydrochloride (10 g, 75 mmol), 13-1 (12 g, 75 mmol) and N-methylmorpholine (9.1 ml, ~2.5 mmol) in N-methyl pyrrolidinone (100 ml) was heated at 120~C forl6 10 h. The solvent was removed at reduced pressure and the residue partitioned between EtOAc (100 ml) and water (50 ml). The organic layer was washed with water and brine (50 ml each) then dried (Na2SO4), filtered and evaporated. The ester 13-2 was isolated as a colorless glass following flash chromatography on silica gel (5%
(~H30H/CH2C12).
I H NMR (300 MHz, CDC13) ~ ~.30 (d, J = 6.8 Hz, 2H), 6.91 (d, J = 6.~s Hz, 2H), 3.~s1 (t, J = 7.1 Hz, 3H), 3.22 (s, 3H), 2.65 (t, J = 7.1 Hz, 2H), 1.41 (s, 9H).
HCI . N~N~ CO2H
3-~(N-Methyl-N'-f4-pyridyl)laminopropionate hydrochloride (13-3) A solution of 13-2 (2.2 g, 9.3 mmol) in 75 ml anhydrous EtOAc was cooled to 0~ and treated with H~l gas for 10 min. The 25 solution was warmed to room temperature and stirred. For 16 h the re.sulting solid was filtered to give 13-3 as a hygroscopic yellow solid.
W 097126250 PCTrUS97/00572 1H NMR (300 mHz, DMSO-d6) ~ 8.26 (d, J = 6.~ Hz, 2H), 7.0 (br d, 2H), 3.~2 (t, J = 7.1 Hz, 2~1), 3.21 ~s, 3H), 2.60 (t, J = 7.1 Hz, 2H).
..
Ni~ O
~N--~N~OEt s 3-~(N-rnethyl-N-(4-pvridyl)laminopropionyl-sarcosine ethyl ester (13-4) Acid 13 3 (383 mg, 1.5 mmol) was coupled with ~arcosine ethyl ester hydrochloride (253 mg, 1.65 mmol) following the EDC/HOBT procedure previously described to give 13-4 as a colorles~
10 glass.
TLC Rf 0.45 (silica, 3~o CH30H/CH2C12) lH NMR (300 MHz, CD~13) ~ ~.21 (d, J = 6.~ Hz, 2H), 6.51 (d, J = 6.g Hz, 2H), 4.20 (q, J - 7.0 Hz, 2H), 4.1~ (,'i, 2H), 3.75 (t, J = 7.0 Hz, 2H), 3.09 (s~ 3H), 3.04 (s, 3H), 2.65 (t, J = 7.0 Hz, 2H), 1.31 (t, J = 7.0 Hz, 15 3H).
Ni~l O
~N ~~ N--b' CH3 CH3 ~
y 3-[(N-Methyl)-N'-(4-pyridyl)~aminopropionyl-sarcosine potassium salt ( l 3-5) A solution of 13 4 (353 mg, 1.26 mmol) in THF (5 ml) wa~;
treated with 0.5 N KOH (2.52 ml, 1.21 mmol) and H20 (5 ml) and stirred at room temperature for 18 h. The solvent was removed i vacuo to afford the potassium salt 13-5 as a white ~olid.
W 097/26250 PCT~US97/00572 1 H NMR (300 MHz, DMSO-d6) ~ ~S.02 (d, J = 6.7 Hz, 2H), 6.57 (d, J =
6.7 Hz, 2H), 3.61 (t, J = 7 Hz, 2H), 3.51 (s, 2H), 2.86 (~, 3H), 2.75 (s, 3H), 2.42 (t, J = 7.0 Hz, 2H).
N~N ~ N~H~ CO2Et 3-[(N-Methyl)-N'-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-,B-alanine ethyl e~ter (13-6) The acid 13-5 wa~ coupled with 7-5 (229 mg, 0.88 mmol) 10 under the E~DC/HOBT procedure to afford 13-6 following chromatography (CH2C12/CH30H/NH2/OH, 90:~:2) lH NMR (300 MHz, CDC13) ~ 8.21 (d, J = 6.Ps Hz, 2H) 7.25 (m, 2H), 6.73 (d, J = 7.0 Hz, lH), 6.51 (d, J = 6.8s Hz, 2H), 4.36 ~m, lH), 61.18 (m, 2H), 3.89 (m, 2H), 3.81 (t, J = 7.0 Hz, 2H), 3.06 (s, 3H), 2.98 (s, 15 3H), 2.85 (m, 2H), 2.65 (m, 2H), 2.51 (m, 2H), 1.83 (m, 2H), 1.20 (m, 3H).
W O 97/26250 PCTrUS97tOO572 N~N~~N~H~,CO2H
CH3 CH3 ~
3-[(N -Methyl)-N'-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethvl)-,13-alanine (13-7) S A solution of the ester 13-6 (75 mg, 0.16 mmol) in THF (S
ml) was treated with 0.5N KOH (320 ml, 0.16 mrnol) and H20 (S ml).
The resulting solution was stirred at room temperature for 7.5 h then evaporated at reduced pressure. The resulting residue was purified by reverse phase a white powder.
1H NMR (300 MHz, DMSO-d6) o 8.61 (d, J = 6.8 Hz, 2H), 7.25 (m, SH), 7.18 (d, J = 7.0 Hz, IH), 6.83 (d, J = 6.8 Hz, 2H), 4.35 (m, lH), 3.~S3 (m, 2H), 3.81 (t, 2H), 3.13 (s, 2H), 2.95 (s, 3H), 2.85 ~m, 2H), 2.65 (m, 2H), 2.56 (m, 2H), 1.86 (m, 2H).
W O 97/26250 PCT~US97/00572 -SC~ME 14 Ph R
11 3 (BOC)20 ~ LiOH BOCN ~,CO2H
BOCN~,CO2EtMeOH/H20 14-2 R = (CH2)2Ph 14-1 14-3 R=H
Ph Ph J 14-4 IR o J
HCI-H2N ~CO2Me BOcN Jl~ N ~CO2Me H
EDC/HOBT/DMF
14-5 R = (CH2)2Ph 1 4-6 R = H
Ph HCI/EtOAc H
HCI-~ - CO2Me R H
14-7 R = (CH2)2Ph 14-3 R=H
WO 97/2625a PCT~US97/00572 ~h BOCN ~,CO2Et N-(t-Butoxycarbonyl)-N-(2-phenylethyl)~lycine ethyl ester (14-1~
The amine 11-3 (1.11 g, ~.36 mmol) and (BOC)2O (1.2g g, 5 5.9 mrnol) in 10 ml T~F were stirred for 48 h under argon. Removal of the solvent in vacuo gave a yellow oil which was purified by chromatography (silica, hexane/EtOAc 9:1) to af~ord 14-1 as a colorless oil.
Rf (silica, hexane/EtOAc 9:1) 0.41.
~h BOC N ~,CO2H
N-(t-Butoxycarbonyl)-N-(2-phenylethyl)glycine (14-2) A solution of the ester 14-1 (1.7 g, 5.5 mmol), 11.1 mL lN
j, - 1 1 9 -SCHEME 13 CONTINUEr) ~.
Ph Nl~'N, ~~N~H - CO2Et KOH
N~ O H - CO
HN--,CO2tBu tert-13utyl 3-N-methylaminopropionate ( 13-1 ) Tert-butyl acrylate (15 g, 1 17 mmol) was added to a solution or methanol saturated with CH3NH2 (300 ml) and stirred at 10 room temperature for 16 h. The solution was evaporated to afford 13-1 as a colorle~s liquid.
W 097/26250 PCT~US97/00572 H NMR (300 MHz, CDC13) ~ 2.81 (t, J = 7.2 Hz, 2H), 2.43 (t, J = 7.2 ~z, 2~), 1.45 (.s, 9~
N~N~ CO2tBu tert-Butyl 3-~(N-methyl-N-(4-pyridyl)laminopropionate (13-2) A mixture of 4-chloropyrid~ne hydrochloride (10 g, 75 mmol), 13-1 (12 g, 75 mmol) and N-methylmorpholine (9.1 ml, ~2.5 mmol) in N-methyl pyrrolidinone (100 ml) was heated at 120~C forl6 10 h. The solvent was removed at reduced pressure and the residue partitioned between EtOAc (100 ml) and water (50 ml). The organic layer was washed with water and brine (50 ml each) then dried (Na2SO4), filtered and evaporated. The ester 13-2 was isolated as a colorless glass following flash chromatography on silica gel (5%
(~H30H/CH2C12).
I H NMR (300 MHz, CDC13) ~ ~.30 (d, J = 6.8 Hz, 2H), 6.91 (d, J = 6.~s Hz, 2H), 3.~s1 (t, J = 7.1 Hz, 3H), 3.22 (s, 3H), 2.65 (t, J = 7.1 Hz, 2H), 1.41 (s, 9H).
HCI . N~N~ CO2H
3-~(N-Methyl-N'-f4-pyridyl)laminopropionate hydrochloride (13-3) A solution of 13-2 (2.2 g, 9.3 mmol) in 75 ml anhydrous EtOAc was cooled to 0~ and treated with H~l gas for 10 min. The 25 solution was warmed to room temperature and stirred. For 16 h the re.sulting solid was filtered to give 13-3 as a hygroscopic yellow solid.
W 097126250 PCTrUS97/00572 1H NMR (300 mHz, DMSO-d6) ~ 8.26 (d, J = 6.~ Hz, 2H), 7.0 (br d, 2H), 3.~2 (t, J = 7.1 Hz, 2~1), 3.21 ~s, 3H), 2.60 (t, J = 7.1 Hz, 2H).
..
Ni~ O
~N--~N~OEt s 3-~(N-rnethyl-N-(4-pvridyl)laminopropionyl-sarcosine ethyl ester (13-4) Acid 13 3 (383 mg, 1.5 mmol) was coupled with ~arcosine ethyl ester hydrochloride (253 mg, 1.65 mmol) following the EDC/HOBT procedure previously described to give 13-4 as a colorles~
10 glass.
TLC Rf 0.45 (silica, 3~o CH30H/CH2C12) lH NMR (300 MHz, CD~13) ~ ~.21 (d, J = 6.~ Hz, 2H), 6.51 (d, J = 6.g Hz, 2H), 4.20 (q, J - 7.0 Hz, 2H), 4.1~ (,'i, 2H), 3.75 (t, J = 7.0 Hz, 2H), 3.09 (s~ 3H), 3.04 (s, 3H), 2.65 (t, J = 7.0 Hz, 2H), 1.31 (t, J = 7.0 Hz, 15 3H).
Ni~l O
~N ~~ N--b' CH3 CH3 ~
y 3-[(N-Methyl)-N'-(4-pyridyl)~aminopropionyl-sarcosine potassium salt ( l 3-5) A solution of 13 4 (353 mg, 1.26 mmol) in THF (5 ml) wa~;
treated with 0.5 N KOH (2.52 ml, 1.21 mmol) and H20 (5 ml) and stirred at room temperature for 18 h. The solvent was removed i vacuo to afford the potassium salt 13-5 as a white ~olid.
W 097/26250 PCT~US97/00572 1 H NMR (300 MHz, DMSO-d6) ~ ~S.02 (d, J = 6.7 Hz, 2H), 6.57 (d, J =
6.7 Hz, 2H), 3.61 (t, J = 7 Hz, 2H), 3.51 (s, 2H), 2.86 (~, 3H), 2.75 (s, 3H), 2.42 (t, J = 7.0 Hz, 2H).
N~N ~ N~H~ CO2Et 3-[(N-Methyl)-N'-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-,B-alanine ethyl e~ter (13-6) The acid 13-5 wa~ coupled with 7-5 (229 mg, 0.88 mmol) 10 under the E~DC/HOBT procedure to afford 13-6 following chromatography (CH2C12/CH30H/NH2/OH, 90:~:2) lH NMR (300 MHz, CDC13) ~ 8.21 (d, J = 6.Ps Hz, 2H) 7.25 (m, 2H), 6.73 (d, J = 7.0 Hz, lH), 6.51 (d, J = 6.8s Hz, 2H), 4.36 ~m, lH), 61.18 (m, 2H), 3.89 (m, 2H), 3.81 (t, J = 7.0 Hz, 2H), 3.06 (s, 3H), 2.98 (s, 15 3H), 2.85 (m, 2H), 2.65 (m, 2H), 2.51 (m, 2H), 1.83 (m, 2H), 1.20 (m, 3H).
W O 97/26250 PCTrUS97tOO572 N~N~~N~H~,CO2H
CH3 CH3 ~
3-[(N -Methyl)-N'-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethvl)-,13-alanine (13-7) S A solution of the ester 13-6 (75 mg, 0.16 mmol) in THF (S
ml) was treated with 0.5N KOH (320 ml, 0.16 mrnol) and H20 (S ml).
The resulting solution was stirred at room temperature for 7.5 h then evaporated at reduced pressure. The resulting residue was purified by reverse phase a white powder.
1H NMR (300 MHz, DMSO-d6) o 8.61 (d, J = 6.8 Hz, 2H), 7.25 (m, SH), 7.18 (d, J = 7.0 Hz, IH), 6.83 (d, J = 6.8 Hz, 2H), 4.35 (m, lH), 3.~S3 (m, 2H), 3.81 (t, 2H), 3.13 (s, 2H), 2.95 (s, 3H), 2.85 ~m, 2H), 2.65 (m, 2H), 2.56 (m, 2H), 1.86 (m, 2H).
W O 97/26250 PCT~US97/00572 -SC~ME 14 Ph R
11 3 (BOC)20 ~ LiOH BOCN ~,CO2H
BOCN~,CO2EtMeOH/H20 14-2 R = (CH2)2Ph 14-1 14-3 R=H
Ph Ph J 14-4 IR o J
HCI-H2N ~CO2Me BOcN Jl~ N ~CO2Me H
EDC/HOBT/DMF
14-5 R = (CH2)2Ph 1 4-6 R = H
Ph HCI/EtOAc H
HCI-~ - CO2Me R H
14-7 R = (CH2)2Ph 14-3 R=H
WO 97/2625a PCT~US97/00572 ~h BOCN ~,CO2Et N-(t-Butoxycarbonyl)-N-(2-phenylethyl)~lycine ethyl ester (14-1~
The amine 11-3 (1.11 g, ~.36 mmol) and (BOC)2O (1.2g g, 5 5.9 mrnol) in 10 ml T~F were stirred for 48 h under argon. Removal of the solvent in vacuo gave a yellow oil which was purified by chromatography (silica, hexane/EtOAc 9:1) to af~ord 14-1 as a colorless oil.
Rf (silica, hexane/EtOAc 9:1) 0.41.
~h BOC N ~,CO2H
N-(t-Butoxycarbonyl)-N-(2-phenylethyl)glycine (14-2) A solution of the ester 14-1 (1.7 g, 5.5 mmol), 11.1 mL lN
15 LiO~I and 11 mL MeOH was stirred at room temperature for 16 h. The mixture was poured into water/EtOAc and acidified with lN HCl to pH
~3. After extraction with EtOAc (2x), the organic layers were washed with brine, dried (MgSO4) and evaporated to give 14-2 as a foam which was u~sed as such in the next step.
W O 97/26250 PCT~US97/00572 Ph Ph ~ O
BOCN 'J~ H ~CO2Me N-[N-(t-Butoxycarbonyl)-N'-(2-phenylethyl)glycyl-3 (R)-(2-phenylethyl)-,B-alanine methyl ester (14-5) The acid 14-5 (502 mg, 1.8 mmol), 3(R)-(2-phenylethyl)-~3-alanine methyl ester hydrochloride 14-4 (see U.S. Patent 5,281,585) (4~2 mg, 2.0 mmol), HOBT (267 mg, 2.0 mmol), EDC hydrochloride (515 mg, 2.7 mmol) and N-methylmorpholine (0.22 ml, 2.0 mmol) were stirred in 10 ml DMF for 16 h under argon. After pouring the 10 solution into EtOAc/10% citric acid (aqueou,~i solution) the mixture was extracted twice with EtOAc, washed with water then brine, dried (MgSO4) and the solvent removed i~l vacuo. The residual yellow oil wa.~; subjected to column chromatography (silica, hexane/EtOAc 1: 1) to give 14-5 as a colorless oil.
Rf (silica, hexane/EtOAc 1:1) 0.44.
Ph o J
HCI-HN ~,J~' N ~CO2Me Ph 14-7 N-[N'-(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-,13-alanine rnethyl ester 20 hydrochloride (14-7) A solution of 14-5 (719 mg, 1.5 mmol) in 40 mL of EtOAc was treated with HCl (g) until saturated. After 30 min the solvent was W O 971262~0 PCT~US97/00572 removed in vacuo and the residue wa,s triturated with ether from which 14-lO crystallized as a white solid.
l H NMR (CD30D) ~ 1.87 (2H, m), 2.5-2.~ (4H, m), 3.05 (2H, m), 3.28 (2H, m), 3.62 (3H, s), 3,7~s (lH, d), 3.~S4 (lH, d), 4.26 (lH, m), 7.1-7.4 5 (lOH, m).
O
BOCN J~ H CO2Me N-[N'-(t-Butoxycarbonyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl 10 ester ~1 4-6) N-(t-butoxycarbonyl)glycine 14-3 (Aldrich) wals coupled with 14-4 according to the procedure described for the preparation of 14-5. The title compound 14 6 was purified by column chromatography (silica, hexane/EtOAc l:l).
15 Rf (silica, hexane/EtOAc l:l) 0.22.
~h O
HCI-H2N ~JI~ N ~ CO2Me N-Gl~cyl-3(R)-(2-phenethyl)-,B-alanine methyl ester hydrochloride 20 (14-~s~
Following the procedure described for the preparation of 14-7 compound 14 6 was converted into 3L4-8.
W O 97/26250 PCTrUS97/00572 IH NMR (CD30D) ~ g (2H, m), 2.5-2.~ (4H, m), 3.64 ~5H, s), 4.25 (lH, m), 7.1-7.3 (SH, m).
W O 97126250 PCTrUS97/OOS72 - 12g-H2N~CO2CH2Ph-0.5 H2SO4 H Me H 11-7 BOCN ~,CO2H
1 4-~ EDC/HOBT/DMF
H ~ MeH HCI/EtOAc BOCN J'N~CO2CH2Ph O Me HCI-H2N ~J~ N ~CO2CH2Ph O Me BOCN~JI'N~CO2CH2Ph s N-[N'-(t-Butoxycarbonyl)glycyl]-3(R)-methyl-~-alanine benzyl e.ster (15-1) N-(t-Butoxycarbonyl)glycine 14-3 (Aldrich) was coupled with 3(R)-methyl-,B-alanine benzyl ester 0.5 ~2SO4 11-7 (Celgene) 10 according to the procedure described for the preparation of 14 5. The title product 15-1 was then obtained by chromatography (silica, hexane/l~tOAc 2:3).
Rf (silica, hexane/EtOAc 1:1) 0.3.
W 097/26250 PCTrUS97/00572 0 Me HCI-H2N ~' H C02cH2Ph l~-(Glycyl-3(R)-methyl-~3-alanine benzyl ester hydrochloride (15-2) S Following the procedure for the preparation of 14-7, compound 15-1 was converted into 15-2 which was isolated as a white solid.
lH NMR (CD30D) o 1.22 (3H, d), 2.5~ (2H, m), 3.53 (lH, d), 3.63 (lH, d), 6.3 (2H, s), 7.35 (SH, m).
W O 97/26250 PCTrUS97/00572 ~ S C~nE~EE l6 ~ ~ \
H2N ~ C 02Et 1)H N ~ C~2 H Cl(16-~) J n-butanol ~~' (Aldrich) 16-1 2) (B O C)20, Et3N, C H2C12 3) LiO H/EtO H/H20 H Cl H2N ~ ~ C 02R2 B O C N ~ ~ C 02H
R CH EDC/HOBT/DMF
15-2 R =Me, 2= 2Ph . R1=(CH2)2Ph, R2=Me ~0~/ H ~ ~ 2 2 16-4 R1=Me, R2=CH2Ph 16-5 R1=(CH2)2Ph, R2=Me 1) LiO H/M e O H/H20 /---\ ~~ ~ N
2) 1 N H Cl \~~-~/ ~ O
16-6 R=Me .16-7 R=(C H2)2Ph ~, .
W O 97/26250 PCT~US97/00572 O ~\H
Ph Ph BOCN~N~ ~ H 2 2) 1 N HCI
PhPh HN N~' N~ ~ CO2H
PhPh BOCN~N~"C02H
~6-3 3-(4-t-Butoxycarbonyl-l-piperizinyl)benzoic acid (16-3) Ethyl 3-aminobenzoate 16-1 (Aldrich, 24.3 g, 0.147 mol) and bi~ (2-chloroethyl)amine hydrochloride 16-2 (Aldrich, 26.3 g, W O 97/26250 PCT~US97/00572 0.147 mol) were heated at reflux in 500 mL n-butanol for 24 h. The solution was concentrated in vacuo, the residue was taken up in EtOAc and washed successively with saturated aqueous NaHCO3 then brine.
After clrying (MgSO4), the solvent was removed and the re,sulting black 5 oil chromatographed (silica, EtOAc then EtOAc/MeOH 1:1 therl MeOH) to give the corresponding piperizine derivative as a mixture of ethyl and butyl esters.
This piperazine (17.8 g, 76 mmol) was dissolved in 500 mL
dry CH2C12 and Et3N (13.3 ml, 95.6 mmol) was then added. To this cooled -5~C solution was added (BOC)2O (17.~ g, 79.9 mmol) in 45 ml dry CH2C12 and stirring was continued until the reaction was complete (as monitored by TLC). The solution was poured into 10% citric acid solution then the organic layer was washed with water, saturated aqueous NaHCO3 and brine. After drying over MgSO4, the solvent was removed in vacuo to give a brown oil. Silica gel chromatography (hexane/EtOAc 1:1) gave the BOC-protected piperazine as a mixture of ethyl cmd butyl esters.
The BOC-protected piperazine (22.1 g) was dissolved in 150 ml 1 N LiOH and 600 ml absolute ethanol and this solution was heated at reflux for 16 h. After removal of the ethanol, EtOAc andlO%
citric acid .solution were added. The organic layer was wa~shed with 1 N
NaOH, the aqueous layer was then re-acidified with lN HCl and extracted with EtOAc. This EtOAc extract was washed with brine, dried (MgSO4) and concentrated to give 16-3 as a white solid.
Rf (silica, hexane/EtOAc 1:1) 0.22.
lH NMR (CDC13) ~ 1.49 (9H, s), 3.21 (4H, br t), 3.61 (4H, br t), 7.16 (lH, dd), 7.36 (lH, t), 7.64 (2H, m).
W O 97/26250 PCTrUS97/00572 BOCN N H
\ ~ H~ ~CO2CH2Ph N- { N'-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl } -3(R)-methyl-~3-alanine benzyl ester (16-4) The acid 16 3 was coupled with I S-2 according to the 5 procedure described for the preparation of 14-5 to yield 16-4.
Rf (silica, EtOAc) 0.45.
HN N~'N~ ~CO2H
10 N- [N'-[3-(1 -Piperazinyl)benzoyl] glycyl] -3 (R)-methyl-~3-alanine trifluoroacetic acid ~alt (16-6) The ester 16 4 (452 mg, O.g4 mmol) was dissolved in 4 ml MeOH, treated with lN LiOH (2.5 ml, 2.5 mmol) and stirred for 48 h.
The solvent was removed under reduced pressure and to the residue was 15 added 10 ml lN HCl. After 10 min, the solution was concentrated and the residue purified !oy preparative HPLC (H20/CH3CN witb 0.1 %
TFA, gradient) to give 16-6.
FAB mass spectrum m/z = 349 (m + 1) lH NMR (CD30D) ~ 1.22 (3H, d), 2.43 (lH, dd), 2.57 (lH, dd), 3.3 20 (4H, m), 3.46 (4H, m), 3.96 (lH, d), 4.04 (lH, d), 4.30 (lH, sextet), 7.25 (IH, m), 7.4 (2H, m), 7.5 (lH, m).
W O 97/26250 PCT~US97/00572 BOCN~N~ H ~ ~CO2Me 1 6-~; Ph N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl~glycyl]-3(R)-(2-phenethyl)-~B-alanine methvl ester (16-5) The acid 16-3 was coupled with 14-P~ according to procedure described for the preparation 14-5 to yield 16-5.
1H NMR (CDCI~) ~ 1.49 (9H, s),1.90 (2H, m), 2.5g (2H, d), 2.63 (2H, m), 3.15 (6H, m), 3.55 (4H, m), 3.62 (3H, s), 4.10 (2H, d), 4.32 (lH, m), 7.0-7.5 (9H, m).
HN N~ N '~' CO2H
1 6-7 P h N-[N'-[3-(1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-~-alanine trifluoroacetic acid salt (16-7) Following the procedure described for the preparation of 16-6,16-5 was converted into 16-7.
FAB mass spectrum m/z = 439 (m+1) Anal. calcd. for C24 H30 N4 O4 ~ 1.35 TFA ~ 1.0 H2O
C, 52.53; H, 5.51; N, 9.18 t 20 found: C, 52.57; H, 5.44; N, 9.26 i' CO2Me 16-8 Ph Ph N-~N'-~3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl] -N'-(2-phenethyl)~lycyll-3(R)-(2-phenethvl)-~-alanine methyl ester (16-2) The acid 16-3 was coupled with 14-7 according to the procedure described for the preparation of 14-5 to yield 16-~s.
Rf (silcia, EtOAc/hexane 2:1) 0.37.
HN N~ N~ CO2H
1 6-g Ph Ph N-[N'-[3-( 1 -Piperazinyl)benzoyl] -N'-(2-phenethyl)glycyl] -3(R)-(2-phenethyl)-~-alanine trifluoroacetic acid salt (16-9) Following the procedure described for the preparation of 16-6, 16-~ was converted into 16-9.
lS FAB ma~ss spectrum m/z = 543 (m + 1) Anal. calcd. for C32 H3~ N4 O4 ~ l.g TFA ~ 0.~ H20 C, 56.09; H, 5.47; N, 7.35 found: C, 56.09; H, 5.41; N, 7.74 W O 971262S0 PCT~US97/00572 BOCN N'[~,CO~H BOCN ~N~lJ NH--c02H
CI-H2N/ C02E,~
~3. After extraction with EtOAc (2x), the organic layers were washed with brine, dried (MgSO4) and evaporated to give 14-2 as a foam which was u~sed as such in the next step.
W O 97/26250 PCT~US97/00572 Ph Ph ~ O
BOCN 'J~ H ~CO2Me N-[N-(t-Butoxycarbonyl)-N'-(2-phenylethyl)glycyl-3 (R)-(2-phenylethyl)-,B-alanine methyl ester (14-5) The acid 14-5 (502 mg, 1.8 mmol), 3(R)-(2-phenylethyl)-~3-alanine methyl ester hydrochloride 14-4 (see U.S. Patent 5,281,585) (4~2 mg, 2.0 mmol), HOBT (267 mg, 2.0 mmol), EDC hydrochloride (515 mg, 2.7 mmol) and N-methylmorpholine (0.22 ml, 2.0 mmol) were stirred in 10 ml DMF for 16 h under argon. After pouring the 10 solution into EtOAc/10% citric acid (aqueou,~i solution) the mixture was extracted twice with EtOAc, washed with water then brine, dried (MgSO4) and the solvent removed i~l vacuo. The residual yellow oil wa.~; subjected to column chromatography (silica, hexane/EtOAc 1: 1) to give 14-5 as a colorless oil.
Rf (silica, hexane/EtOAc 1:1) 0.44.
Ph o J
HCI-HN ~,J~' N ~CO2Me Ph 14-7 N-[N'-(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-,13-alanine rnethyl ester 20 hydrochloride (14-7) A solution of 14-5 (719 mg, 1.5 mmol) in 40 mL of EtOAc was treated with HCl (g) until saturated. After 30 min the solvent was W O 971262~0 PCT~US97/00572 removed in vacuo and the residue wa,s triturated with ether from which 14-lO crystallized as a white solid.
l H NMR (CD30D) ~ 1.87 (2H, m), 2.5-2.~ (4H, m), 3.05 (2H, m), 3.28 (2H, m), 3.62 (3H, s), 3,7~s (lH, d), 3.~S4 (lH, d), 4.26 (lH, m), 7.1-7.4 5 (lOH, m).
O
BOCN J~ H CO2Me N-[N'-(t-Butoxycarbonyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl 10 ester ~1 4-6) N-(t-butoxycarbonyl)glycine 14-3 (Aldrich) wals coupled with 14-4 according to the procedure described for the preparation of 14-5. The title compound 14 6 was purified by column chromatography (silica, hexane/EtOAc l:l).
15 Rf (silica, hexane/EtOAc l:l) 0.22.
~h O
HCI-H2N ~JI~ N ~ CO2Me N-Gl~cyl-3(R)-(2-phenethyl)-,B-alanine methyl ester hydrochloride 20 (14-~s~
Following the procedure described for the preparation of 14-7 compound 14 6 was converted into 3L4-8.
W O 97/26250 PCTrUS97/00572 IH NMR (CD30D) ~ g (2H, m), 2.5-2.~ (4H, m), 3.64 ~5H, s), 4.25 (lH, m), 7.1-7.3 (SH, m).
W O 97126250 PCTrUS97/OOS72 - 12g-H2N~CO2CH2Ph-0.5 H2SO4 H Me H 11-7 BOCN ~,CO2H
1 4-~ EDC/HOBT/DMF
H ~ MeH HCI/EtOAc BOCN J'N~CO2CH2Ph O Me HCI-H2N ~J~ N ~CO2CH2Ph O Me BOCN~JI'N~CO2CH2Ph s N-[N'-(t-Butoxycarbonyl)glycyl]-3(R)-methyl-~-alanine benzyl e.ster (15-1) N-(t-Butoxycarbonyl)glycine 14-3 (Aldrich) was coupled with 3(R)-methyl-,B-alanine benzyl ester 0.5 ~2SO4 11-7 (Celgene) 10 according to the procedure described for the preparation of 14 5. The title product 15-1 was then obtained by chromatography (silica, hexane/l~tOAc 2:3).
Rf (silica, hexane/EtOAc 1:1) 0.3.
W 097/26250 PCTrUS97/00572 0 Me HCI-H2N ~' H C02cH2Ph l~-(Glycyl-3(R)-methyl-~3-alanine benzyl ester hydrochloride (15-2) S Following the procedure for the preparation of 14-7, compound 15-1 was converted into 15-2 which was isolated as a white solid.
lH NMR (CD30D) o 1.22 (3H, d), 2.5~ (2H, m), 3.53 (lH, d), 3.63 (lH, d), 6.3 (2H, s), 7.35 (SH, m).
W O 97/26250 PCTrUS97/00572 ~ S C~nE~EE l6 ~ ~ \
H2N ~ C 02Et 1)H N ~ C~2 H Cl(16-~) J n-butanol ~~' (Aldrich) 16-1 2) (B O C)20, Et3N, C H2C12 3) LiO H/EtO H/H20 H Cl H2N ~ ~ C 02R2 B O C N ~ ~ C 02H
R CH EDC/HOBT/DMF
15-2 R =Me, 2= 2Ph . R1=(CH2)2Ph, R2=Me ~0~/ H ~ ~ 2 2 16-4 R1=Me, R2=CH2Ph 16-5 R1=(CH2)2Ph, R2=Me 1) LiO H/M e O H/H20 /---\ ~~ ~ N
2) 1 N H Cl \~~-~/ ~ O
16-6 R=Me .16-7 R=(C H2)2Ph ~, .
W O 97/26250 PCT~US97/00572 O ~\H
Ph Ph BOCN~N~ ~ H 2 2) 1 N HCI
PhPh HN N~' N~ ~ CO2H
PhPh BOCN~N~"C02H
~6-3 3-(4-t-Butoxycarbonyl-l-piperizinyl)benzoic acid (16-3) Ethyl 3-aminobenzoate 16-1 (Aldrich, 24.3 g, 0.147 mol) and bi~ (2-chloroethyl)amine hydrochloride 16-2 (Aldrich, 26.3 g, W O 97/26250 PCT~US97/00572 0.147 mol) were heated at reflux in 500 mL n-butanol for 24 h. The solution was concentrated in vacuo, the residue was taken up in EtOAc and washed successively with saturated aqueous NaHCO3 then brine.
After clrying (MgSO4), the solvent was removed and the re,sulting black 5 oil chromatographed (silica, EtOAc then EtOAc/MeOH 1:1 therl MeOH) to give the corresponding piperizine derivative as a mixture of ethyl and butyl esters.
This piperazine (17.8 g, 76 mmol) was dissolved in 500 mL
dry CH2C12 and Et3N (13.3 ml, 95.6 mmol) was then added. To this cooled -5~C solution was added (BOC)2O (17.~ g, 79.9 mmol) in 45 ml dry CH2C12 and stirring was continued until the reaction was complete (as monitored by TLC). The solution was poured into 10% citric acid solution then the organic layer was washed with water, saturated aqueous NaHCO3 and brine. After drying over MgSO4, the solvent was removed in vacuo to give a brown oil. Silica gel chromatography (hexane/EtOAc 1:1) gave the BOC-protected piperazine as a mixture of ethyl cmd butyl esters.
The BOC-protected piperazine (22.1 g) was dissolved in 150 ml 1 N LiOH and 600 ml absolute ethanol and this solution was heated at reflux for 16 h. After removal of the ethanol, EtOAc andlO%
citric acid .solution were added. The organic layer was wa~shed with 1 N
NaOH, the aqueous layer was then re-acidified with lN HCl and extracted with EtOAc. This EtOAc extract was washed with brine, dried (MgSO4) and concentrated to give 16-3 as a white solid.
Rf (silica, hexane/EtOAc 1:1) 0.22.
lH NMR (CDC13) ~ 1.49 (9H, s), 3.21 (4H, br t), 3.61 (4H, br t), 7.16 (lH, dd), 7.36 (lH, t), 7.64 (2H, m).
W O 97/26250 PCTrUS97/00572 BOCN N H
\ ~ H~ ~CO2CH2Ph N- { N'-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl } -3(R)-methyl-~3-alanine benzyl ester (16-4) The acid 16 3 was coupled with I S-2 according to the 5 procedure described for the preparation of 14-5 to yield 16-4.
Rf (silica, EtOAc) 0.45.
HN N~'N~ ~CO2H
10 N- [N'-[3-(1 -Piperazinyl)benzoyl] glycyl] -3 (R)-methyl-~3-alanine trifluoroacetic acid ~alt (16-6) The ester 16 4 (452 mg, O.g4 mmol) was dissolved in 4 ml MeOH, treated with lN LiOH (2.5 ml, 2.5 mmol) and stirred for 48 h.
The solvent was removed under reduced pressure and to the residue was 15 added 10 ml lN HCl. After 10 min, the solution was concentrated and the residue purified !oy preparative HPLC (H20/CH3CN witb 0.1 %
TFA, gradient) to give 16-6.
FAB mass spectrum m/z = 349 (m + 1) lH NMR (CD30D) ~ 1.22 (3H, d), 2.43 (lH, dd), 2.57 (lH, dd), 3.3 20 (4H, m), 3.46 (4H, m), 3.96 (lH, d), 4.04 (lH, d), 4.30 (lH, sextet), 7.25 (IH, m), 7.4 (2H, m), 7.5 (lH, m).
W O 97/26250 PCT~US97/00572 BOCN~N~ H ~ ~CO2Me 1 6-~; Ph N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl~glycyl]-3(R)-(2-phenethyl)-~B-alanine methvl ester (16-5) The acid 16-3 was coupled with 14-P~ according to procedure described for the preparation 14-5 to yield 16-5.
1H NMR (CDCI~) ~ 1.49 (9H, s),1.90 (2H, m), 2.5g (2H, d), 2.63 (2H, m), 3.15 (6H, m), 3.55 (4H, m), 3.62 (3H, s), 4.10 (2H, d), 4.32 (lH, m), 7.0-7.5 (9H, m).
HN N~ N '~' CO2H
1 6-7 P h N-[N'-[3-(1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-~-alanine trifluoroacetic acid salt (16-7) Following the procedure described for the preparation of 16-6,16-5 was converted into 16-7.
FAB mass spectrum m/z = 439 (m+1) Anal. calcd. for C24 H30 N4 O4 ~ 1.35 TFA ~ 1.0 H2O
C, 52.53; H, 5.51; N, 9.18 t 20 found: C, 52.57; H, 5.44; N, 9.26 i' CO2Me 16-8 Ph Ph N-~N'-~3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl] -N'-(2-phenethyl)~lycyll-3(R)-(2-phenethvl)-~-alanine methyl ester (16-2) The acid 16-3 was coupled with 14-7 according to the procedure described for the preparation of 14-5 to yield 16-~s.
Rf (silcia, EtOAc/hexane 2:1) 0.37.
HN N~ N~ CO2H
1 6-g Ph Ph N-[N'-[3-( 1 -Piperazinyl)benzoyl] -N'-(2-phenethyl)glycyl] -3(R)-(2-phenethyl)-~-alanine trifluoroacetic acid salt (16-9) Following the procedure described for the preparation of 16-6, 16-~ was converted into 16-9.
lS FAB ma~ss spectrum m/z = 543 (m + 1) Anal. calcd. for C32 H3~ N4 O4 ~ l.g TFA ~ 0.~ H20 C, 56.09; H, 5.47; N, 7.35 found: C, 56.09; H, 5.41; N, 7.74 W O 971262S0 PCT~US97/00572 BOCN N'[~,CO~H BOCN ~N~lJ NH--c02H
CI-H2N/ C02E,~
17-2 /2. deprotection HN N'~ ~ CO2H
1 7-4 ~3N
1, HCI-H2N ~co 2. deprotection r~N
HN N~l \IH~NH ~CO2H
W O 97/26250 PCTrUS97/00572 - 13~
BOCN ~N ~,~ N ~C02H
3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl ~Iycine (17- 1) The acid 16-3 was coupled with glycine ethyl ester followed 5 by hydrolysis of the resulting ester using previously described chemistry to yield 17- 1.
lH NMR (300 MHz, CD30D) o 1.4~ (9H, .s), 3.~ (4H, m), 3.59 (4H, m), 4.08 (2H, s), 7.22 (lH, m), 7.40 (2H, m), 7.55 (lH, s).
A O
HN N~ N ~ N CO2H
N-[N'-~3-(1 -Piperazinyl)benzoyl]glycyl] -3 (S)-ethynyl-13-alanine trifluoroacetic acid salt (17-4) The acid 17 1 was coupled with 3(S)-ethynyl-,13-alanine ethyl ester hydrochloride (Zablocki et al., J. Med. Chem., 1995, 38, 15 2378-2394) using standard peptide coupling conditions. The product was then fully deprotected using previously de~cribed methodology to give, after reverse phase chromotography, 17-4 as the trifluoroacetate salt.
FAB mass spectrum m/z - 359 (M+1) 20 Anal. calculated for C 1 gH22N4O4- 1.10 TFA-0.30 H20 C, 49.59; H, 4.~; N, 11.45 Found: C, 49.~8; H, 4.~0, N, 11.57 W O 97/26250 PCT~US97/00572 ~ O
HN N~ N~N CO2H
N-{ N''-[3-( 1 -Piperazinyl)benzoyl~glycyl ~-3(S)-(3-pyridyl)-~B-alanine tri~luoroacetic acid salt (17-5) The acid,l7-1 was coupled with 3(S)-(3-pyridyl)-~-al~nine 5 ethyl ester hydrochloride (Rico et al., J. Org Chem., 1993, vol. 58, p.
794g) using standard peptide coupling condition,~i. The product was then fully deprotected using previously described methodology to give, after rever~e phase chromatography, 17-5 a.s the trifluoroacetate salt.
PAB mass spectrum m/z = 412 (M+l) Anal. calculated for C21H2sNsO4-2.55 TFA-0.75 H2O
C, 43.gO; H, 4.09; N, 9.79 Found: C, 43.76; H, 3.9~; N, 10.15 NO2~CO2H H2 NH2~CO2H
10% Pd-C MeOH ~F
1 7-4 ~3N
1, HCI-H2N ~co 2. deprotection r~N
HN N~l \IH~NH ~CO2H
W O 97/26250 PCTrUS97/00572 - 13~
BOCN ~N ~,~ N ~C02H
3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl ~Iycine (17- 1) The acid 16-3 was coupled with glycine ethyl ester followed 5 by hydrolysis of the resulting ester using previously described chemistry to yield 17- 1.
lH NMR (300 MHz, CD30D) o 1.4~ (9H, .s), 3.~ (4H, m), 3.59 (4H, m), 4.08 (2H, s), 7.22 (lH, m), 7.40 (2H, m), 7.55 (lH, s).
A O
HN N~ N ~ N CO2H
N-[N'-~3-(1 -Piperazinyl)benzoyl]glycyl] -3 (S)-ethynyl-13-alanine trifluoroacetic acid salt (17-4) The acid 17 1 was coupled with 3(S)-ethynyl-,13-alanine ethyl ester hydrochloride (Zablocki et al., J. Med. Chem., 1995, 38, 15 2378-2394) using standard peptide coupling conditions. The product was then fully deprotected using previously de~cribed methodology to give, after reverse phase chromotography, 17-4 as the trifluoroacetate salt.
FAB mass spectrum m/z - 359 (M+1) 20 Anal. calculated for C 1 gH22N4O4- 1.10 TFA-0.30 H20 C, 49.59; H, 4.~; N, 11.45 Found: C, 49.~8; H, 4.~0, N, 11.57 W O 97/26250 PCT~US97/00572 ~ O
HN N~ N~N CO2H
N-{ N''-[3-( 1 -Piperazinyl)benzoyl~glycyl ~-3(S)-(3-pyridyl)-~B-alanine tri~luoroacetic acid salt (17-5) The acid,l7-1 was coupled with 3(S)-(3-pyridyl)-~-al~nine 5 ethyl ester hydrochloride (Rico et al., J. Org Chem., 1993, vol. 58, p.
794g) using standard peptide coupling condition,~i. The product was then fully deprotected using previously described methodology to give, after rever~e phase chromatography, 17-5 a.s the trifluoroacetate salt.
PAB mass spectrum m/z = 412 (M+l) Anal. calculated for C21H2sNsO4-2.55 TFA-0.75 H2O
C, 43.gO; H, 4.09; N, 9.79 Found: C, 43.76; H, 3.9~; N, 10.15 NO2~CO2H H2 NH2~CO2H
10% Pd-C MeOH ~F
NH2~ 3 steps . ( W O 97/26250 PCTrUS97/00572 SCHEME l ~s (CONT'D) 11 / \
1. HCI-H2N~ ~ \
2. deprotection 1,1 HN ~N~ NH~NH~~-- 2 ~N
1 . H2N CO2Et 2. deprotection ~N
HN N,~, O C02H
t8-6 W 097126250 PCT~US97/00572 H2N ~C02H
5-Amino-2-fluorobenzoic acid 1~-2 2-Fluoro-5-nitrobenzoic acid 1~-1 (Aldrich) was reduced using 10% Pd-C catalyst in MeOH under an atmosphere of H2 to give, 5 after filtration of the catalyst and removal of the solvent, 1~-2 as a ~iolid. R~= 0.54 ~silica, 10-1-1 EtOH-NH40H-H20) H2N ~NH CO2Me N-(5-Amino-2-Fluorobenzoyl)~lycine methvl ester lg-3 The acid 18-2 was coupled with glycine methyl e~ster using standard peptide coupling conditions to give 1 g-3.
Rf = 0.65 (silica; EtOAc/MeOH 9:1) BOCN~N~ H CO2H
15 5-~4-t-Butoxycarbonyl-l-piperizinyl)-2-fluoro benzoyl glycine lg-4 Following the procedure described for the preparation of 16-3, the aniline 1~-3 was converted into the piperazine-acid 1~s-4 lH NMR (300 MHz, CD30D) ~ 1.46 (9H, s), 3.1 1 (4H, m), 3.5~ (4H, m), 4.12 (2H, s), 7.05-7.21 (2H, m), 7.40 (lH, m).
W O 97/26250 PCTrUS97/00572 CF3COz (~) H2N~N,~C,~ N /~ N--CO ~H
~8-5 H
N- { N'-~2-Fluoro-5-(1 -piperazinyl)benzoyl]glycyl ~ -3(S)-ethynyl-,13-alanine trifluoro acetic acicl salt 1~-5 Following the procedure described for the preparation of 17-4, compound 18-4 was converted into l ~-5.
FAB mas~s spectrum m/z = 377 (M+l) Anal. calculated for C1 ~¢H21 N404F- 1.30TFA-0.50 H20 C, 46.37; H, 4.40; N, 10.50 Found: C, 46.34; H, 4.37; N, 10.58 C F3C02 (~) H2N N ~F ~
N- { N'-[2-fluoro-5-(1 -piperazinyl)benzoyl]glycyl ~ -3 (S)-(3 -pyridyl)~
alanine trifluoroacetic acid salt 1~-6 Following the procedure descri7Oed for the preparation of 17-5, compound 1~-4 was converted into 1 g-6.
FAB mass spectrum m/z = 430 (M+l) Analysis calculated for C21H24N504F-2.65 TFA-0.9OH20 C, 42.24; H, 3.83; N, 9.37 Found: C, ~2.25; H, 3.81; N, 9.71 W O 97/262~a PCTAUS97/00572 N~NH2 CHO ~ CO2Et HCI-H2N~CO2t-Bu L-Proline, EtOH, ~
~! Cbz-Glycine ,N~ CO2Et BOP, NMM
O
H2, Pd/C, EtOAc CbzN~Jl~H CO2t-Bu H
~N CO2Et 19-7 19-4 H2, Pd/C, EtOAc;
HCI, Et2O
6 N HCI, 50~C
H O
HCI~ C02H HCI-H2N J~ NH ~ CO2t-Bu BOP, NMI\/I, CH3CN
I
W O 97/26250 PCT~US97/00~72 SCHEME 19 CONT'D
H H
N ~J~ N ~\,CO2t-Bu TFA, CH2CI2 ~N R N ,CO2H
N~ ~ CO2Et s Ethyl 4-(l~X-naphthyridin-2-yl)butanoate (19-3) Aminoaldehyde 19 1 (2.02 g, 16.6 mmol, prepared according to Het. 1993, 36, 2513), ketone 19-2 (5.3 mL, 33.1 mmol) and L-proline (0.48 g, 4.17 mmol) were combined in 75 mL EtOH.
10 After heating at reflux overnight the reaction wa,s concentrated. Flash chromatography (silica, EtOAc) provided 19-3 ~s an off-white crystalline ,solid.
TLC Rf 0.23 (silica, EtOAc) lH NMR (300 MHz, CDC13): ~ 9.09 (dd, J=4, 2Hz, lH), ~S.17 (dd, J=8, 2Hz, lH), 8.12 (d, J=8Hz, lH), 7.46 (dd, J=8, 4Hz, lH), 7.42 (d, J=8Hz, w o 97/2625a~ PCTrUS97/00572 lH), 4.12 (~1, J=7Hz, 2H), 3.11 (t, J=~sHz, 2H), 2.44 (t, J=7Hz, lH), 2.26 (~ln, J=~Hz, 2H), 1.25 (t, J=7Hz, 3H).
CO2Et Ethyl 4-( l .2~3~4-tetrahydro- 1 .8-naphthyridin-7-vl)butanoate ( 19-4) A solution of 19-3 (2.3 g, 9.4 mmol) in 50 mL EtOAc wa,~;
treated with 10% Pd/C (230 mg) and a hydrogen balloon. After 4 d the reaction filtered through celite, concentrated, and purified by flash chromatography (silica, 70% EtOAc/hexane), providing 19-4 as a yellow oil.
TLC Rf 0.40 (silica, EtOAc) H NMR (300 MHz, CDC13): ~ 7.05 (d, J=7Hz, IH), 6.35 (d, J=7Hz, lH), ~.73 (br s, lH), 4.12 (4, J=7Hz, 2H), 2.69 (t, J=6Hz, 2H), 2.57 (t, J=~sHz, 2H), 2.33 (t, J=7Hz, 2H), l.9~s (m, 2H), 1.90 (m, 2H), 1.25 (t, J=7H:z, 3H).
HCI~ ~'CO2H
4-(1,".,3,4-Tetrahydro-l,~¢-naphthyridin-7-yl)butanoic acid hydrochloride (19-5) 4 Ester 19-4 (1.8 g, 7.25 mmol) in 36 mL 6 N HCl was heated at 50~C for 4 h, then concentrated, providing 19-5 as a yellow solid.
W 097/26250 PCT~US97/00572 lH NMR (300 MHz, CD30D): ~ 7.59 (d, J=7Hz, IH), 6.63 (d, J=7Hz, lH), 3.50 (t, J=5Hz, 2H), 2.82 (t, J=6Hz, 2H), 2.74 (t, J=8Hz, 2H), 2.38 (t, J=7Hz, 2H), 2.02-1.90 (m, 4H).
IH o CbzN~JI~ H CO2t-Bu N-Cbz-Glycyl-~-alanine t-butyl e~ster (19-7) N-CBz-Glycine (1.0 g, 4.7~s mmol), amine 19-6 (0.91 g, 5.02 mrnol), NMM (2.1 mL, 19.1 mmol) and BOP ~3.17 g, 7.17 mmol) were combined in 15 mL DMF. After stirring overnight the mixture wa,s concentrated, diluted with EtOAc, washed with water, sat.
NaHCO3, water, 5% KHSO4 and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 60% EtOAc/hexane) provided 19-7 as a colorless oil.
TLC Rf 0.24 (silica, 60% EtOAc/hexane) 1 H NMR (400 MHz, d6-DMSO): ~ 7.~s9 (br t, J=5Hz, lH), 7.44 (br t, J=6Hz, lH), 7.40-7.30 (m, 5H), 5.02 (.s, 2H), 3.56 (d, J=6Hz, 2E~), 3.25 (q, J=6Hz, 2H), 2.35 (t, J=7Hz, 2H), 1.40 (~;, 9H).
HCI-H2N ~Jl' H CO2t-Bu Glycyl-~3-alanine t-butyl ester hydrochloride (19-~
A solution of 19-7 (1.51 g, 4.49 mmol) in 40 mL EtOAc wa.s treated with ~0% Pd/C (0.30 g), and a H2 balloon. After stirring overnight under a hydrogen atmosphere, an additional 200 mg of 10%
W 097/2625~ rcTrusg7/oo572 Pd/C wa~ added and hydrogenation was continued for 4 h before filtering through Celite and concentrating, providing the free amine as a colorle~ oil. The amine was dissolved in Et2O and an exces.~i of 1 M
HCI in Et2O was added. ~oncentration provided 19-~ as a waxy solid.
lH NMR (free am~ne, 400 MHz, d6-DMSO): ~ 8.31 (br s, lH), 5.30 (br s, 2H), 3.29 (~, J=6Hz, 2H), 3.25 (s, 2H), 2.38 (t, J=7Hz, 2H), 1.41 (s, 9H).
H H
~NJI~N~CO2t-Bu t9-9 4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)bu~anoyl-glycyl-~-alanine t-but~l ester (19-9) A mixture of 19-5 (62 mg, 0.24 mmol), 19-~ (69 mg, 0.29 mmoll), NMM (130 !uL, 1.2 mmol) and BOP (160 mg, 0.36 mmol) in 2 mL CH3CN was stirred overnight. After diluting with EtOAc the mixture was washed with sat. NaHCO3, water (5x) and brine, dried (MgSO4), filtered and concentrated, providing 19 9.
TLC Rf 0.79 (silica, 25% NH3-sat. EtOH/EtOAc) lH NMR (300 MHz, CDC13): ~ 8.50 (br t, lH), 7.08 (d, J=7Hz, lH), 6.64 (br t, lH), 6.33 (d, J=7Hz, lH), 5.69 (br s, lH), 3.99 (d, J=7Hz, 2H), 3.53 (q, J=6Hz, 2H), 3.43 (m, 2H), 2.69 (t, J=6Hz, 2H), 2.60 (t, J=7Hz, 2H), 2.46 (t, J=6Hz, 2H), 2.25 (t, J=7Hz, 2H), 2.05-1.90 (m, 4H), 1.45 (s, 9H).
W O 97/26250 PCT~US97/OOS72 - 14~s -H H
N Jl' N ~\,CO2H
1~-10 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-~-alanine (19-10) Ester 19-9 (69 mg, 0.17 mmol) was di.~solved in 1 mL
5 CH2C12 at 0~C, 1 mL TFA was added, and the reaction wa~ warrned to ambient temperature for 6 hr. After concentrating and azeotroping with toluene, fla~h chromatography (silica, 7:20:1:1 EtO~c/EtOH/H2O/NH4OlI) provided 1 9-10 as a white solid.
TLC Rf 0.38 (silica, 7:20:1:1 EtOAc/EtOH/H2O/NH4OH) lH NMR (400 MHz, D20): ~ 7.53 (d, J=7Hz, lH), 6.59 (d, J=7Hz, lH), 3.~5 (s, 2H), 3.46 (t, J=6Hz, 2H), 3.42 (t, J=7Hz, 2H), 2.78 (t, J=6Hz, 2H), 2.72 (t, J=8Hz, 2H), 2.40 (apparent q, J=7Hz, 4H), 2.00 (qn, J=6Hz, 2H), 1.92 (qn, J=6Hz, 2H).
W ~ 971Z625~ PCT~US97/00572 SCHE~ME 20 HCI~ " ~ H
1. HCI-H2N~ ~ \
2. deprotection 1,1 HN ~N~ NH~NH~~-- 2 ~N
1 . H2N CO2Et 2. deprotection ~N
HN N,~, O C02H
t8-6 W 097126250 PCT~US97/00572 H2N ~C02H
5-Amino-2-fluorobenzoic acid 1~-2 2-Fluoro-5-nitrobenzoic acid 1~-1 (Aldrich) was reduced using 10% Pd-C catalyst in MeOH under an atmosphere of H2 to give, 5 after filtration of the catalyst and removal of the solvent, 1~-2 as a ~iolid. R~= 0.54 ~silica, 10-1-1 EtOH-NH40H-H20) H2N ~NH CO2Me N-(5-Amino-2-Fluorobenzoyl)~lycine methvl ester lg-3 The acid 18-2 was coupled with glycine methyl e~ster using standard peptide coupling conditions to give 1 g-3.
Rf = 0.65 (silica; EtOAc/MeOH 9:1) BOCN~N~ H CO2H
15 5-~4-t-Butoxycarbonyl-l-piperizinyl)-2-fluoro benzoyl glycine lg-4 Following the procedure described for the preparation of 16-3, the aniline 1~-3 was converted into the piperazine-acid 1~s-4 lH NMR (300 MHz, CD30D) ~ 1.46 (9H, s), 3.1 1 (4H, m), 3.5~ (4H, m), 4.12 (2H, s), 7.05-7.21 (2H, m), 7.40 (lH, m).
W O 97/26250 PCTrUS97/00572 CF3COz (~) H2N~N,~C,~ N /~ N--CO ~H
~8-5 H
N- { N'-~2-Fluoro-5-(1 -piperazinyl)benzoyl]glycyl ~ -3(S)-ethynyl-,13-alanine trifluoro acetic acicl salt 1~-5 Following the procedure described for the preparation of 17-4, compound 18-4 was converted into l ~-5.
FAB mas~s spectrum m/z = 377 (M+l) Anal. calculated for C1 ~¢H21 N404F- 1.30TFA-0.50 H20 C, 46.37; H, 4.40; N, 10.50 Found: C, 46.34; H, 4.37; N, 10.58 C F3C02 (~) H2N N ~F ~
N- { N'-[2-fluoro-5-(1 -piperazinyl)benzoyl]glycyl ~ -3 (S)-(3 -pyridyl)~
alanine trifluoroacetic acid salt 1~-6 Following the procedure descri7Oed for the preparation of 17-5, compound 1~-4 was converted into 1 g-6.
FAB mass spectrum m/z = 430 (M+l) Analysis calculated for C21H24N504F-2.65 TFA-0.9OH20 C, 42.24; H, 3.83; N, 9.37 Found: C, ~2.25; H, 3.81; N, 9.71 W O 97/262~a PCTAUS97/00572 N~NH2 CHO ~ CO2Et HCI-H2N~CO2t-Bu L-Proline, EtOH, ~
~! Cbz-Glycine ,N~ CO2Et BOP, NMM
O
H2, Pd/C, EtOAc CbzN~Jl~H CO2t-Bu H
~N CO2Et 19-7 19-4 H2, Pd/C, EtOAc;
HCI, Et2O
6 N HCI, 50~C
H O
HCI~ C02H HCI-H2N J~ NH ~ CO2t-Bu BOP, NMI\/I, CH3CN
I
W O 97/26250 PCT~US97/00~72 SCHEME 19 CONT'D
H H
N ~J~ N ~\,CO2t-Bu TFA, CH2CI2 ~N R N ,CO2H
N~ ~ CO2Et s Ethyl 4-(l~X-naphthyridin-2-yl)butanoate (19-3) Aminoaldehyde 19 1 (2.02 g, 16.6 mmol, prepared according to Het. 1993, 36, 2513), ketone 19-2 (5.3 mL, 33.1 mmol) and L-proline (0.48 g, 4.17 mmol) were combined in 75 mL EtOH.
10 After heating at reflux overnight the reaction wa,s concentrated. Flash chromatography (silica, EtOAc) provided 19-3 ~s an off-white crystalline ,solid.
TLC Rf 0.23 (silica, EtOAc) lH NMR (300 MHz, CDC13): ~ 9.09 (dd, J=4, 2Hz, lH), ~S.17 (dd, J=8, 2Hz, lH), 8.12 (d, J=8Hz, lH), 7.46 (dd, J=8, 4Hz, lH), 7.42 (d, J=8Hz, w o 97/2625a~ PCTrUS97/00572 lH), 4.12 (~1, J=7Hz, 2H), 3.11 (t, J=~sHz, 2H), 2.44 (t, J=7Hz, lH), 2.26 (~ln, J=~Hz, 2H), 1.25 (t, J=7Hz, 3H).
CO2Et Ethyl 4-( l .2~3~4-tetrahydro- 1 .8-naphthyridin-7-vl)butanoate ( 19-4) A solution of 19-3 (2.3 g, 9.4 mmol) in 50 mL EtOAc wa,~;
treated with 10% Pd/C (230 mg) and a hydrogen balloon. After 4 d the reaction filtered through celite, concentrated, and purified by flash chromatography (silica, 70% EtOAc/hexane), providing 19-4 as a yellow oil.
TLC Rf 0.40 (silica, EtOAc) H NMR (300 MHz, CDC13): ~ 7.05 (d, J=7Hz, IH), 6.35 (d, J=7Hz, lH), ~.73 (br s, lH), 4.12 (4, J=7Hz, 2H), 2.69 (t, J=6Hz, 2H), 2.57 (t, J=~sHz, 2H), 2.33 (t, J=7Hz, 2H), l.9~s (m, 2H), 1.90 (m, 2H), 1.25 (t, J=7H:z, 3H).
HCI~ ~'CO2H
4-(1,".,3,4-Tetrahydro-l,~¢-naphthyridin-7-yl)butanoic acid hydrochloride (19-5) 4 Ester 19-4 (1.8 g, 7.25 mmol) in 36 mL 6 N HCl was heated at 50~C for 4 h, then concentrated, providing 19-5 as a yellow solid.
W 097/26250 PCT~US97/00572 lH NMR (300 MHz, CD30D): ~ 7.59 (d, J=7Hz, IH), 6.63 (d, J=7Hz, lH), 3.50 (t, J=5Hz, 2H), 2.82 (t, J=6Hz, 2H), 2.74 (t, J=8Hz, 2H), 2.38 (t, J=7Hz, 2H), 2.02-1.90 (m, 4H).
IH o CbzN~JI~ H CO2t-Bu N-Cbz-Glycyl-~-alanine t-butyl e~ster (19-7) N-CBz-Glycine (1.0 g, 4.7~s mmol), amine 19-6 (0.91 g, 5.02 mrnol), NMM (2.1 mL, 19.1 mmol) and BOP ~3.17 g, 7.17 mmol) were combined in 15 mL DMF. After stirring overnight the mixture wa,s concentrated, diluted with EtOAc, washed with water, sat.
NaHCO3, water, 5% KHSO4 and brine, dried (MgSO4), filtered and concentrated. Flash chromatography (silica, 60% EtOAc/hexane) provided 19-7 as a colorless oil.
TLC Rf 0.24 (silica, 60% EtOAc/hexane) 1 H NMR (400 MHz, d6-DMSO): ~ 7.~s9 (br t, J=5Hz, lH), 7.44 (br t, J=6Hz, lH), 7.40-7.30 (m, 5H), 5.02 (.s, 2H), 3.56 (d, J=6Hz, 2E~), 3.25 (q, J=6Hz, 2H), 2.35 (t, J=7Hz, 2H), 1.40 (~;, 9H).
HCI-H2N ~Jl' H CO2t-Bu Glycyl-~3-alanine t-butyl ester hydrochloride (19-~
A solution of 19-7 (1.51 g, 4.49 mmol) in 40 mL EtOAc wa.s treated with ~0% Pd/C (0.30 g), and a H2 balloon. After stirring overnight under a hydrogen atmosphere, an additional 200 mg of 10%
W 097/2625~ rcTrusg7/oo572 Pd/C wa~ added and hydrogenation was continued for 4 h before filtering through Celite and concentrating, providing the free amine as a colorle~ oil. The amine was dissolved in Et2O and an exces.~i of 1 M
HCI in Et2O was added. ~oncentration provided 19-~ as a waxy solid.
lH NMR (free am~ne, 400 MHz, d6-DMSO): ~ 8.31 (br s, lH), 5.30 (br s, 2H), 3.29 (~, J=6Hz, 2H), 3.25 (s, 2H), 2.38 (t, J=7Hz, 2H), 1.41 (s, 9H).
H H
~NJI~N~CO2t-Bu t9-9 4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)bu~anoyl-glycyl-~-alanine t-but~l ester (19-9) A mixture of 19-5 (62 mg, 0.24 mmol), 19-~ (69 mg, 0.29 mmoll), NMM (130 !uL, 1.2 mmol) and BOP (160 mg, 0.36 mmol) in 2 mL CH3CN was stirred overnight. After diluting with EtOAc the mixture was washed with sat. NaHCO3, water (5x) and brine, dried (MgSO4), filtered and concentrated, providing 19 9.
TLC Rf 0.79 (silica, 25% NH3-sat. EtOH/EtOAc) lH NMR (300 MHz, CDC13): ~ 8.50 (br t, lH), 7.08 (d, J=7Hz, lH), 6.64 (br t, lH), 6.33 (d, J=7Hz, lH), 5.69 (br s, lH), 3.99 (d, J=7Hz, 2H), 3.53 (q, J=6Hz, 2H), 3.43 (m, 2H), 2.69 (t, J=6Hz, 2H), 2.60 (t, J=7Hz, 2H), 2.46 (t, J=6Hz, 2H), 2.25 (t, J=7Hz, 2H), 2.05-1.90 (m, 4H), 1.45 (s, 9H).
W O 97/26250 PCT~US97/OOS72 - 14~s -H H
N Jl' N ~\,CO2H
1~-10 4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-~-alanine (19-10) Ester 19-9 (69 mg, 0.17 mmol) was di.~solved in 1 mL
5 CH2C12 at 0~C, 1 mL TFA was added, and the reaction wa~ warrned to ambient temperature for 6 hr. After concentrating and azeotroping with toluene, fla~h chromatography (silica, 7:20:1:1 EtO~c/EtOH/H2O/NH4OlI) provided 1 9-10 as a white solid.
TLC Rf 0.38 (silica, 7:20:1:1 EtOAc/EtOH/H2O/NH4OH) lH NMR (400 MHz, D20): ~ 7.53 (d, J=7Hz, lH), 6.59 (d, J=7Hz, lH), 3.~5 (s, 2H), 3.46 (t, J=6Hz, 2H), 3.42 (t, J=7Hz, 2H), 2.78 (t, J=6Hz, 2H), 2.72 (t, J=8Hz, 2H), 2.40 (apparent q, J=7Hz, 4H), 2.00 (qn, J=6Hz, 2H), 1.92 (qn, J=6Hz, 2H).
W ~ 971Z625~ PCT~US97/00572 SCHE~ME 20 HCI~ " ~ H
20-1a HCI ~ ~OH + H I O
BOP
NMM
N ~ N OEt MeOH
1 N NaOH
W 097/26250 PCT~US97/00572 N~ N OH
1) H2NCH2CO2Bn, BOP, NMM, C H3CN
2) H2, 10% Pd/C, ~tOH
~I N~J~ N/~OH
20-la 4~ 2.3.4-Tetrahydro-1.~-naphthyridin-7-yl)butanoyl glycine (20-1a) A mixture of l9 5 (1.02 g, 4.0 mmol), glycine benzyl ester (0.8 g, 4.0 mmol), NMM (1.76 ml, 16 mmol) and BOP ~2.03 g, 4.6 mmol) in CH3CN (100 ml) was stirred overnight. The reaction was concentrated and the residue was partitioned between EtOAc and H2O.
10 The organic layer wa~ washed with sat. NaHCO3 solution, brine, dried (MgSO4), filtered and concentrated to a yellow gum which was purified by flash chromatography (~ilica, 1:1, acetone/CH2C12) to provide the ester as a colorless gum.
~ solution of the ester (1.3 g, 3.5 mmol) in EtOH (100 ml) 15 was hydrogenated at 1 atm for lg hr. The reaction was diluted with EtOAc (200 ml) to dissolve the product, filtered and concentrated to a solid which was sonicated with ether ( 100 ml) to provide 20-la as a colorless solid.
W O 97J262~0 PCTrUS97/00572 TLC lRf 0.35 (~ilica, EtOH/NH3) lH NMR (300 MHz, CD30D): o 7.50 (d, J=7Hz, 2H), 6.59 (d, J=7Hz, 2H), 3.80 (s, 2H), 3.47 (t, J=6Hz, 2H), 2.79 (t, J=6Hz, 2H), 2.72 (t, J=7~:z, 2H), 2.26 (t, J=7Hz, 2H), 2.02 (m, 2H), 1.94 (m, 2H).
4-(1 " ,3,4-Tetrahydro- 1,~-naphthyridin-7 -yl)butanoyl-glycyl-3 (S)-pyridin-3-yl-,B-alanine ethyl ester (20-2~
The CH3CN solution (300 mL) of 20-la (164 mg, 0.59 mmol), 20-1 (Rico et al., J. Org. Chem., 1993, 58, 7948) (15~ mg, 0.58 10 mmol), NMM (260 ,ul, 2.36 mlnol) and BOP (300 mg, 0.6~ mmol) was stirred under ambient conditions for 48 h. The reaction was concentrated to a yellow gum which was purified by flash chromatography (silica, 9:1 CH2C12/EtOH~NH3) to provide 20-2 .lS a colorless gum.
Rf 0.71 (.~ilica, 9: 1 CH2C12/EtOH~NH3) lH NMR (300 MHz, CD30D): ~ 8.53 (bs, lH), 8.42 (d, J=5Hz, lH), 7.82 (d, J=gHz, lH), 7.39 (dd, J=8Hz, 5Hz, lH), 7.10 (d, J=8Hz, lH), 6.36 (d, J=7Hz, lH), 5.40 (t, J=7Hz, lH), 4.07 (q, d=7Hz, 2H), 3.g5 (~
2El), 3.36 (t, J=6Hz, 2H), 2.91 (m, 2H), 2.6g (t, J=6Hz, 2H), 2.51 (t, 20 J=7H:z, 2H), 2.23 (t, J=7Hz, 2H), 1.89 (m, 4H), 1.16 (t, J=7Hz, 3H).
4-( l ,r' ,3,4-Tetrahydro- 1,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-pyridin-3-yl-~-alanine (20-3) A methanol ~solution (10 mL) of 20-2 (190 mg, 0.42 mmol) 25 and lN NaOH (2.1 mL, 2.1 mmol) was stirred under ambient conditions for 1~ h. The reaction was concentrated to dryness and the residue neutralized with lN HCI and the resultant solution concentrated to a gum which was chromatographed (silica, 38/1/1 EtOH/NH40H/H20) to provide a solid which was purified by HPLC using a VyOAC C18 semi 30 prep ~olumn with gradient elution [95:5~99.9:0.1 H2O/TFA)/(99.9:0.1 CH3CN/TFA) ~ 50:50 (99.9:0.1 H2O/TFA)/(99.9:0.1 CH3CN/TFA)80 min] to provide 20-3 as a hygroscopic solid ditrifluoroacetate salt.
Rf 0.36 (silica 38: 1: 1 EtOH/NH40H/EI20) lH NMR (300 MHz, ~D30D): ~ ~.79 (bs, lH), ~s.65 (d, J=SHz, IH), 8.7 (d, J=8Hz, IH), 7.84 (m, lH), 7.57 (d, J=7Hz, lH), 6.61 (d, J=7Hz, lH), 5.44 (t, J=7Hz, 4H), 3.88 (m, 2H), 3.48 (t, J-5Hz, 2H), 2.98 (d, J=7Hz, 2H), 2.81 (t, J=6Hz, 2H), 2.70 (m, 2H), 2.31 (m, 2H), 1.96 (m, 4H).
s W 097126250 PCTrUS97/00572 ,, N~CI
HN~ }CO2~t NMP, NMM
1 10~ ~NH2 N /~N ~CO2Et BrCH2CO2tBu, Et3N, CH3CN
BOP
NMM
N ~ N OEt MeOH
1 N NaOH
W 097/26250 PCT~US97/00572 N~ N OH
1) H2NCH2CO2Bn, BOP, NMM, C H3CN
2) H2, 10% Pd/C, ~tOH
~I N~J~ N/~OH
20-la 4~ 2.3.4-Tetrahydro-1.~-naphthyridin-7-yl)butanoyl glycine (20-1a) A mixture of l9 5 (1.02 g, 4.0 mmol), glycine benzyl ester (0.8 g, 4.0 mmol), NMM (1.76 ml, 16 mmol) and BOP ~2.03 g, 4.6 mmol) in CH3CN (100 ml) was stirred overnight. The reaction was concentrated and the residue was partitioned between EtOAc and H2O.
10 The organic layer wa~ washed with sat. NaHCO3 solution, brine, dried (MgSO4), filtered and concentrated to a yellow gum which was purified by flash chromatography (~ilica, 1:1, acetone/CH2C12) to provide the ester as a colorless gum.
~ solution of the ester (1.3 g, 3.5 mmol) in EtOH (100 ml) 15 was hydrogenated at 1 atm for lg hr. The reaction was diluted with EtOAc (200 ml) to dissolve the product, filtered and concentrated to a solid which was sonicated with ether ( 100 ml) to provide 20-la as a colorless solid.
W O 97J262~0 PCTrUS97/00572 TLC lRf 0.35 (~ilica, EtOH/NH3) lH NMR (300 MHz, CD30D): o 7.50 (d, J=7Hz, 2H), 6.59 (d, J=7Hz, 2H), 3.80 (s, 2H), 3.47 (t, J=6Hz, 2H), 2.79 (t, J=6Hz, 2H), 2.72 (t, J=7~:z, 2H), 2.26 (t, J=7Hz, 2H), 2.02 (m, 2H), 1.94 (m, 2H).
4-(1 " ,3,4-Tetrahydro- 1,~-naphthyridin-7 -yl)butanoyl-glycyl-3 (S)-pyridin-3-yl-,B-alanine ethyl ester (20-2~
The CH3CN solution (300 mL) of 20-la (164 mg, 0.59 mmol), 20-1 (Rico et al., J. Org. Chem., 1993, 58, 7948) (15~ mg, 0.58 10 mmol), NMM (260 ,ul, 2.36 mlnol) and BOP (300 mg, 0.6~ mmol) was stirred under ambient conditions for 48 h. The reaction was concentrated to a yellow gum which was purified by flash chromatography (silica, 9:1 CH2C12/EtOH~NH3) to provide 20-2 .lS a colorless gum.
Rf 0.71 (.~ilica, 9: 1 CH2C12/EtOH~NH3) lH NMR (300 MHz, CD30D): ~ 8.53 (bs, lH), 8.42 (d, J=5Hz, lH), 7.82 (d, J=gHz, lH), 7.39 (dd, J=8Hz, 5Hz, lH), 7.10 (d, J=8Hz, lH), 6.36 (d, J=7Hz, lH), 5.40 (t, J=7Hz, lH), 4.07 (q, d=7Hz, 2H), 3.g5 (~
2El), 3.36 (t, J=6Hz, 2H), 2.91 (m, 2H), 2.6g (t, J=6Hz, 2H), 2.51 (t, 20 J=7H:z, 2H), 2.23 (t, J=7Hz, 2H), 1.89 (m, 4H), 1.16 (t, J=7Hz, 3H).
4-( l ,r' ,3,4-Tetrahydro- 1,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-pyridin-3-yl-~-alanine (20-3) A methanol ~solution (10 mL) of 20-2 (190 mg, 0.42 mmol) 25 and lN NaOH (2.1 mL, 2.1 mmol) was stirred under ambient conditions for 1~ h. The reaction was concentrated to dryness and the residue neutralized with lN HCI and the resultant solution concentrated to a gum which was chromatographed (silica, 38/1/1 EtOH/NH40H/H20) to provide a solid which was purified by HPLC using a VyOAC C18 semi 30 prep ~olumn with gradient elution [95:5~99.9:0.1 H2O/TFA)/(99.9:0.1 CH3CN/TFA) ~ 50:50 (99.9:0.1 H2O/TFA)/(99.9:0.1 CH3CN/TFA)80 min] to provide 20-3 as a hygroscopic solid ditrifluoroacetate salt.
Rf 0.36 (silica 38: 1: 1 EtOH/NH40H/EI20) lH NMR (300 MHz, ~D30D): ~ ~.79 (bs, lH), ~s.65 (d, J=SHz, IH), 8.7 (d, J=8Hz, IH), 7.84 (m, lH), 7.57 (d, J=7Hz, lH), 6.61 (d, J=7Hz, lH), 5.44 (t, J=7Hz, 4H), 3.88 (m, 2H), 3.48 (t, J-5Hz, 2H), 2.98 (d, J=7Hz, 2H), 2.81 (t, J=6Hz, 2H), 2.70 (m, 2H), 2.31 (m, 2H), 1.96 (m, 4H).
s W 097126250 PCTrUS97/00572 ,, N~CI
HN~ }CO2~t NMP, NMM
1 10~ ~NH2 N /~N ~CO2Et BrCH2CO2tBu, Et3N, CH3CN
LiOH, THF/H20 ~7 HN ~,CO2tBu N~ N ~ CO2H
' ~' PYCLU, iPr2NEt, DMF
SC~IEME 21 (CONT'D) HCI, EtOAc N~ N~ N~, CO2tBu o N~ N~_~ N~,CO2H
~HCI 21-5 HCI H2N~CO2Et PYCLU, iPr2NEt, DMF
..
~ o 1,1 N~N/ > ~N~N~CO2Et Y ~ H -W O 97/2625~1 PCTrUS97/00572 N ~ N ~ C O2Et Ethyl N-pyridin-4-yli~onipecotate (21- l ) Ethyl isonipecotate (6.0 g, 38.66 mmol), 4-chloropyridine hydrochloride (5.9 g, 38.66 mrnol) and N-methylmorpholine (9.3 mL, 5 85.0 rnmol) were dissolved in N-methylpyrrolidinone (50 mL) and the resulting solution heated at 100~ for 4~s h. The ~olution was concentrated in vacuo and t~e residue dissolved in ethyl acetate (200 mL), washed with water and brine (2 x 100 mL), then dried (Na2SO4) and evaporated. The resulting residue was purified by flash chromatography (5%MeOH/CH2C12) to afford 21-1 as a crystalline ~solid.
lH NMR (300 MHz, CDCl3) ~i 8.21 (d, J=6.8Hz, 2H), 6.78 (d, J=6.gHz, 2H), 4.18 (q, J-7.0Hz, 2H), 3.85 (m, 2H), 3.10 (m, 2H), 2.61 (m, lH) 2.05 (m, 2H), 1.~5 (m, 2H), 1.23 (t, J=7.0Hz, 3H).
N~ N~ C02H
N-P~ridin-4-ylisonipecotic acid (21-2) A solution of ester 21-1 (10 g, 42.7 mmol) in THF (50 mL) was ~reated with lN LiOH (47 mL, 47.0 mmol) and H2O (50 mL). The 20 resulting solution was concentrated and the aqueous residue cooled to 0~C, then adjusted to pH 6 with lN H~l and the re~ulting solid 21-2, collected by filtration.
lH NMR (300 MHz, D20) ~ 7.95 (d, 6.8Hz, 2H), 6.73 (d, 6.~Hz, 2H), 3.76 (d, J=12.PsHz, 2H), 2.~1 (m, 2H), 2.20 (m, lH), 1.~5 (d, J=12.gHz, - 25 2H), 1.55 (m, 2H).
W O 97t262~0 PCTrUS97/OOS72 HN ~CO2tBu tert-Butyl-N-cvclopropyl Iycine (21-3) A mixture of cyclopropylamine (10.0 g, 175.1 mmol) and triethylamine (4.9 ml, 35.5 mmol) in 100 ml CH2Cl2 was cooled to 0~
5 and treated with tert-butyl bromoacetate (5.25 ml, 35.0 mmol). The resulting mixture was stirred at 0~C for 2 h, refluxed for 1.5 h, then cooled and washed with .sat. NaHC03, and brine (50 ml each) then dried (Na2S04) and evaporated to afford 21-3 a colorless oil.
lH NMR (300 MHz, CDC13) ~ 3.35 (s, 2H), 2.19 (m, lH), 2.0~ (br s, lH), 1.4g (s, 9H), 0.47 (m, 2H), 0.3~ (m, 2H).
N~N~N~CO2tBu tert-Butvl N-pyridin-4-vlisonipecotyl-N-cyclopropyl~lycine (21-4) A solution of acid 21-2 (500 mg, 2.36 mmol), ester 21-4 (404 mg, 2.36 mmol), chloro-N,N,N',N'-bis(pentamethylene)-folmamidinium hexafluorophosphate (PYCLU) (g51 mg, 2.36 mmol), and diisopropylethyl amine (305 mg, 2.36 mmol) in anhydrous DMF
(50 mL) was stirred at room temperature for 18 h then concentrated in vacuo to afford a yellow residue. Chromatography on silica gel (l: l 2{) MeOH/EtOAc) afforded 21-4 as a crystalline solid.
lH NMR (300 MHz, CD30D) o 8.12 (d, J=6.X Hz, 2H), 6.75 (d, 3=6.~Hz, 2H), 3.94 (d, J=12.gHz, 2H), 3.~5 (.s, 2H), 2.gl (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 1.55 (m, 2H), 1.42 (s, 9H), 0.47 (m, 2H), 0.3g (m, 2H).
W O 97/262~0 PCTAUS97/00572 ., N~N ~ N~,C02H
~HCI 21-5 N-Pyridin-4-ylisonipecotyl-N-cyclopropylglvcyl (21-5~
Ester 21-5 (250 mg, 0.70 mmol) was su.spended in EtOAc (25 mL), cooled to 0~ and treated with HCI gas for 15 min. The 5 resulting solution was stirred at 0~ for 3.5 h then evaporated to give 21-5 ;3s a yellow glass.
lH NMR (300 MHz, CD30D) o 8.18 (d, J=6.8Hz, 2H), 7.1~ (d, J-6.8Hz, 2H), 4.24 (d, J=12.8Hz, 2H), 3.95 (s, 2H), 3.21 (m, 2H), 1.95 (m, 2H), l.g5 ~m, 2H), 1.62 (m, 2H), 0.g7 (m, 2H), 0.75 (m, 2H).
N~N~ ~I'N~' 2 Ethyl N-pyridin-4-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-,B-alalline (21-7) A solution of acid 21 -5 (232 mg, 0.68 mrnol), ester 21 -6 (121 ~ng, 0.68 mmol) (21-6 prepared as described in U.S. patent 5,272,162), PYCLU (245 mg, 0.68 mmol), and diisopropylethyl amine (176 mg, 0.68 mmol) in anhydrous DMF ~50 ml) was stirred at room temperature for 18 h then concentrated i~l vacuo to afford a yellow re.sidue. Preparative reverse phase chromatographic purification 20 affordied ester 21-7 as its TFA salt.
lH NMR (300 MlIz, CD30D) ~ X.48 (d, J=6.8Hz, lH), 8.08 (d, J=6.81Hz, 2H), 7.18 (d, J=6.8Hz, 2H), 5.01 (m, H), 4.24 (d, J=12.~sHz, 2H), ~.12 (~1, J=7Hz, 2H), 3.99 (s, 2H), 3.72 (m, lH), 3.31 (m, 2H), 2.95 W O 97/26250 PCTrUS97/00572 - 15~¢ -(m, lH), 2.73 (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 1.21 (t, J=7.0Hz, 2H), 0.87 (m, 2H), 0.75 (m, 2H).
~ 0 11 N~ N~}~ N~J~ N~ CO2H
o S N-Pyridin-4-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl~
alanine ~21-8) A solution of ester 21-7 (180 mg, 0.422 mmol), in THF (10 mL) was treated with lN LiOH (O.P~4 mL, O.g4 mmol) and stirred at room temperature for 16 h. The mixture was concentrated and the 10 residue purified by preparative reverse phase chromatography to afford 19-~S as its TFA salt.
lH NMR (300 MHz, CD30D) ~ 8.40 (d, J=6.XHz, lH), 8.21 (d, J=6.~Hz, 2H), 7.21 (d, J=6.~sHz, 2H), 4.81 (m, lH), 4.22 (d, J=12.8Hz, 2H), 3.99 (m, 2H), 3.72 (m, lH), 3.31 (m, 2H), 2.95 (m, lH), 2.76 (m, lH), 2.71 (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 0.87 (m, 2H), 0.75 (m, 2H).
W O 97n6250 PCT~US97/00~72 <CO2Et N~GI NMP, NMM ~ ~CO2Et ~ 22-1 LiOH, THF/H20 <CO2H y N~N ~> HN ~CO2tBu ~2~2 21-3 PYCLU, iPr2NEt, DMF
W 097/26250 PCT~US97/00572 SCHEME 22 (CONT'D) N~N/ > ~ HCI, EtOAc \~,N~CO2tBu ~-3 ~N~,C02H
~HCI
' ~' PYCLU, iPr2NEt, DMF
SC~IEME 21 (CONT'D) HCI, EtOAc N~ N~ N~, CO2tBu o N~ N~_~ N~,CO2H
~HCI 21-5 HCI H2N~CO2Et PYCLU, iPr2NEt, DMF
..
~ o 1,1 N~N/ > ~N~N~CO2Et Y ~ H -W O 97/2625~1 PCTrUS97/00572 N ~ N ~ C O2Et Ethyl N-pyridin-4-yli~onipecotate (21- l ) Ethyl isonipecotate (6.0 g, 38.66 mmol), 4-chloropyridine hydrochloride (5.9 g, 38.66 mrnol) and N-methylmorpholine (9.3 mL, 5 85.0 rnmol) were dissolved in N-methylpyrrolidinone (50 mL) and the resulting solution heated at 100~ for 4~s h. The ~olution was concentrated in vacuo and t~e residue dissolved in ethyl acetate (200 mL), washed with water and brine (2 x 100 mL), then dried (Na2SO4) and evaporated. The resulting residue was purified by flash chromatography (5%MeOH/CH2C12) to afford 21-1 as a crystalline ~solid.
lH NMR (300 MHz, CDCl3) ~i 8.21 (d, J=6.8Hz, 2H), 6.78 (d, J=6.gHz, 2H), 4.18 (q, J-7.0Hz, 2H), 3.85 (m, 2H), 3.10 (m, 2H), 2.61 (m, lH) 2.05 (m, 2H), 1.~5 (m, 2H), 1.23 (t, J=7.0Hz, 3H).
N~ N~ C02H
N-P~ridin-4-ylisonipecotic acid (21-2) A solution of ester 21-1 (10 g, 42.7 mmol) in THF (50 mL) was ~reated with lN LiOH (47 mL, 47.0 mmol) and H2O (50 mL). The 20 resulting solution was concentrated and the aqueous residue cooled to 0~C, then adjusted to pH 6 with lN H~l and the re~ulting solid 21-2, collected by filtration.
lH NMR (300 MHz, D20) ~ 7.95 (d, 6.8Hz, 2H), 6.73 (d, 6.~Hz, 2H), 3.76 (d, J=12.PsHz, 2H), 2.~1 (m, 2H), 2.20 (m, lH), 1.~5 (d, J=12.gHz, - 25 2H), 1.55 (m, 2H).
W O 97t262~0 PCTrUS97/OOS72 HN ~CO2tBu tert-Butyl-N-cvclopropyl Iycine (21-3) A mixture of cyclopropylamine (10.0 g, 175.1 mmol) and triethylamine (4.9 ml, 35.5 mmol) in 100 ml CH2Cl2 was cooled to 0~
5 and treated with tert-butyl bromoacetate (5.25 ml, 35.0 mmol). The resulting mixture was stirred at 0~C for 2 h, refluxed for 1.5 h, then cooled and washed with .sat. NaHC03, and brine (50 ml each) then dried (Na2S04) and evaporated to afford 21-3 a colorless oil.
lH NMR (300 MHz, CDC13) ~ 3.35 (s, 2H), 2.19 (m, lH), 2.0~ (br s, lH), 1.4g (s, 9H), 0.47 (m, 2H), 0.3~ (m, 2H).
N~N~N~CO2tBu tert-Butvl N-pyridin-4-vlisonipecotyl-N-cyclopropyl~lycine (21-4) A solution of acid 21-2 (500 mg, 2.36 mmol), ester 21-4 (404 mg, 2.36 mmol), chloro-N,N,N',N'-bis(pentamethylene)-folmamidinium hexafluorophosphate (PYCLU) (g51 mg, 2.36 mmol), and diisopropylethyl amine (305 mg, 2.36 mmol) in anhydrous DMF
(50 mL) was stirred at room temperature for 18 h then concentrated in vacuo to afford a yellow residue. Chromatography on silica gel (l: l 2{) MeOH/EtOAc) afforded 21-4 as a crystalline solid.
lH NMR (300 MHz, CD30D) o 8.12 (d, J=6.X Hz, 2H), 6.75 (d, 3=6.~Hz, 2H), 3.94 (d, J=12.gHz, 2H), 3.~5 (.s, 2H), 2.gl (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 1.55 (m, 2H), 1.42 (s, 9H), 0.47 (m, 2H), 0.3g (m, 2H).
W O 97/262~0 PCTAUS97/00572 ., N~N ~ N~,C02H
~HCI 21-5 N-Pyridin-4-ylisonipecotyl-N-cyclopropylglvcyl (21-5~
Ester 21-5 (250 mg, 0.70 mmol) was su.spended in EtOAc (25 mL), cooled to 0~ and treated with HCI gas for 15 min. The 5 resulting solution was stirred at 0~ for 3.5 h then evaporated to give 21-5 ;3s a yellow glass.
lH NMR (300 MHz, CD30D) o 8.18 (d, J=6.8Hz, 2H), 7.1~ (d, J-6.8Hz, 2H), 4.24 (d, J=12.8Hz, 2H), 3.95 (s, 2H), 3.21 (m, 2H), 1.95 (m, 2H), l.g5 ~m, 2H), 1.62 (m, 2H), 0.g7 (m, 2H), 0.75 (m, 2H).
N~N~ ~I'N~' 2 Ethyl N-pyridin-4-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-,B-alalline (21-7) A solution of acid 21 -5 (232 mg, 0.68 mrnol), ester 21 -6 (121 ~ng, 0.68 mmol) (21-6 prepared as described in U.S. patent 5,272,162), PYCLU (245 mg, 0.68 mmol), and diisopropylethyl amine (176 mg, 0.68 mmol) in anhydrous DMF ~50 ml) was stirred at room temperature for 18 h then concentrated i~l vacuo to afford a yellow re.sidue. Preparative reverse phase chromatographic purification 20 affordied ester 21-7 as its TFA salt.
lH NMR (300 MlIz, CD30D) ~ X.48 (d, J=6.8Hz, lH), 8.08 (d, J=6.81Hz, 2H), 7.18 (d, J=6.8Hz, 2H), 5.01 (m, H), 4.24 (d, J=12.~sHz, 2H), ~.12 (~1, J=7Hz, 2H), 3.99 (s, 2H), 3.72 (m, lH), 3.31 (m, 2H), 2.95 W O 97/26250 PCTrUS97/00572 - 15~¢ -(m, lH), 2.73 (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 1.21 (t, J=7.0Hz, 2H), 0.87 (m, 2H), 0.75 (m, 2H).
~ 0 11 N~ N~}~ N~J~ N~ CO2H
o S N-Pyridin-4-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl~
alanine ~21-8) A solution of ester 21-7 (180 mg, 0.422 mmol), in THF (10 mL) was treated with lN LiOH (O.P~4 mL, O.g4 mmol) and stirred at room temperature for 16 h. The mixture was concentrated and the 10 residue purified by preparative reverse phase chromatography to afford 19-~S as its TFA salt.
lH NMR (300 MHz, CD30D) ~ 8.40 (d, J=6.XHz, lH), 8.21 (d, J=6.~Hz, 2H), 7.21 (d, J=6.~sHz, 2H), 4.81 (m, lH), 4.22 (d, J=12.8Hz, 2H), 3.99 (m, 2H), 3.72 (m, lH), 3.31 (m, 2H), 2.95 (m, lH), 2.76 (m, lH), 2.71 (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 0.87 (m, 2H), 0.75 (m, 2H).
W O 97n6250 PCT~US97/00~72 <CO2Et N~GI NMP, NMM ~ ~CO2Et ~ 22-1 LiOH, THF/H20 <CO2H y N~N ~> HN ~CO2tBu ~2~2 21-3 PYCLU, iPr2NEt, DMF
W 097/26250 PCT~US97/00572 SCHEME 22 (CONT'D) N~N/ > ~ HCI, EtOAc \~,N~CO2tBu ~-3 ~N~,C02H
~HCI
HCI H2N CO2Et PYCI U, iPr2NEt, DMF
N~ ~,N~J~ N~CO2Et 2f ~N~ Z
wo 97126250 PCTAUS97/00572 C02Et N~ N~
l~thyl N-pyridin-4-vlnipecotate (22-1~
Ethyl (~) nipecotate (7.0 g, 44.53 mmol) was reacted with 4-chlorpyridine hydrochloride (6.67 g, 44.53 mmol) as described for 5 21-1 t:o give the title compound as a yellow ~olid.
lH NMR (300 MHz, CDC13) ~ ~.22 (d, J=6.~Hz, 2H), 6.68 (d, J=6.8Hz, 2H), 4.18 (4, J=7.0Hz, 2H), 3.85 (m, lH), 3.72 (m, lH), 3.21 (m, lH), 3.10 (m, lH), 2.60 (m, lH), 2.08 (m, lH), l.gl (m, 2H), 1.60 (m, lH), 1.13 (t, J=7.0Hz, 3H).
~ ~<
N~ N~
N-Pyridin-4-ylnipecotic acid ~22-2) P~epared from 22-1 (764 mg, 3.25 mmol) in a manner similar to that described f~or 21-2.
lH NMR (300 MHz, DMSO-d6) ~ 8.13 (d, J=6.8Hz, 2H), 6.74 (d, J=6.8Hz, 2H), 4.08 (d, lH), 3.78 (m, lH), 2.92 (m, 2H), 2.10 (m, lH), 1.95 (m, lH), 1.71 (m, lH), 1.42 (m, 2H).
W O 97/26250 PCT~US97100572 N3 2f N~,CO2 Bu tert-Butyl N-pyridin-4-ylnipecotyl-N-cyclopropyl~lycine (22-3) Prepared from 22-2 (320 mg, 1.51 mmol) and 21 -3 (258 mg, l.Sl mrnol) in a manner similar to that described for 21-4.
lH NMR (300 MHz, CDC13) ~ 8.12 (d, 6.8Hz, 2H), 6.62 (d, J=6.8Hz, 2H), 3.94 (s, 2H), 3.85 (m, lH), 3.12 (m, lH), 3.08 (m, lH), 251 (m, 2H), 1.95 (m, lH), 1.85 (m, 2H), 1.58 (m, 2H), 1.42 (s, 9H), 0.47 (m, 2H), 0.38 (m, 2H).
N~ N
~HCI
o N-Pyridin-4-ylnipecotyl-N-cyclopropyl~lycine hydrochloride (22-4) Ester 22-3 (250 mg, 0.70 mmol) was suspended in EtOAc (25 mL), cooled to 0~ and treated with HCI gas for 15 min. The resulting solution was stir~ed for 3.5 h then evaporated to give 22-4 as a white solid.
1H NMR (300 MHz, DMSO-d6) ~ 8 18 (d, J=6.8Hz, 2H), 7.1~ (d, J=6.8Hz, 2H), 4.24 (d, J=12.8Hz, 2H), 3.95 (m, 2H), 3.21 (m, IH), 1.94 20 (m, lH), 1.85 (m, lH), 1.72 (m, lH), 1.53 (m, lH), 0.87 (m, 2H), 0.75 (m, 2H).
~ N~ N~ CO~Et Ethy~ N-pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~B-alanine (22-5) Prepared from 22-4 (195 mg, 0.60 mmol) in a manner 5 similar to that described ~or 21-7.
lH NMR (300 MHz, CD30D) ~ 8.49 (d, J=6.~Hz, IH), ~s.17 (d, J=6.~Hz, 2H), 7.21 (d, J=6.8Hz, 2H), 5.15 (m, lH), 4.26 (m, IH), 4.21 (d, lH), 4.08 (q, 2H), 3.82 (m, lH), 3.5-3.3 (m, 3H), 2.95 (m, lH), 2.76 (m, lH), 2.71 (m, 2H), 2.1~ (m, lH), 1.95 (m, lH), 1.~1 (m, lH), 1.72 (m, lH), 1.21 (t~ 3H), O.g7 (m, 2H).
21' ~ N~-- 2 N-Pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-alanine (22-6~
Prepared from 22-4 (20 mg, 0.04 mmol) in a manner 15 simil.~r to that described for 21-8.
FAB mass spectrum m/z = 399 (M + 1).
lH NMR (300 MHz, CD30D) ~ 8.16 (d, J=6Hz, 2H), 6.91 (d, J=6.gHz, 2H), 5.05 (m, lH), 4.26 (d, Hz, lH), 4.21 (d, lH), 3.~s2 (m, lh), 3.5-3.3 (m, 3H), 2.95 (m, lH), 2.76 (m, lH), 2.71 (m, 2H), 2.15 (m, IH), 1.95 (m, lH), l.~sl (m, lH), 1.72 (m, lH), 1.21 (t, lH), 0.87 (m, 2H).
W O 97/262~0 PCTrUS97/00572 CH3 NaN(TMS)2 ~j~ TH F, -78~C N>=~
N N Br/ CO2CH3 N~/\--CO2CH3 1 N NaOH
EtO H
N~9 HCl-HN~C02Et ~==<
N ~ CO2H
BOP, NMM, DMF ~y N~ f ~OEt10 /O Pd/C
W O 97/262~,0 PCTAUS97/00572 - 1~5 -SCHEME 23 (CONT'D) HN/~ ~ 0 1N NaOH
OEt Z3 ~ ~
IJ ~~
HN/ > ~7 0 HCI-H2N~--C~2Et N~ N~J~ BOP, NMM
2q,-6 HN~> ~ H 1. 1N NaOH
N = ~N JINH~CO2Et EtOH
O 2. Prep HPLC
H~ N7~1 H'~~co H
W 097/26250 PCTrUS97/00572 N~
N~--c02CH3 Methyl 4-(l.g-naphthyridin-4-yl)butyrate (23-2) To a stirred solution of naphthyridine 23-1 (Hamada, Y. ~t al., Chenl. Pharm. Bull. Soc., 1971, 19(9), 1857-1862), (2.2 g, 15.2 S rnmol) and THF (200 ml) at -7~~C was added NaN(TMS)2 (lM/THF, l~s ml, l ~S mmol) dropwi.se over a 20 min period. After 30 minutes at -7~~C, methyl 3-bromopropionate was added in a stream. After 30 min, the reaction was (luenched with 50 ml 10% KHSO4. The mixture was extracted with Et2O. The rem~ining aqueous portion was basified with 10 sat. NaHCO3 and then extracted with EtOAc. The EtOAc portion was washed with brine, dried (MgSO4) and concentrated. Flash chromotography (silica, 2% EtOH/EtOAc) gave the ester 23-2 (1.61 g) as a yellow oil.
TLC Rf = 0.27 (silica, 2% EtOH/EtOAc) lH NMR (400 MHz, CDC13) ~ 9.14 (m, lH), 9.35 (d, J=4Hz, lH), g.50 (d, J=7Hz, lH), 7.52 (~1, J=4Hz, IH), 7.33 (d, J=4Hz, lH), 3.71 (s, 3H), 3.14 (t, J=~Hz, 2H), 2.46 (t, J=7Hz, 2H), 2.09 (m, 2H).
N ~ .
N~CO2H
20 4-(l.g-Naphthyridin-4-yl)butanoic acid (1-3) A solution of ester 23-2 (1.60 g, 6.9 mmol), lN NaOH (7 ml, 7 mmol) and EtOH (20 ml) was stirred at ambient temperature for 1.0 h. The solution was extracted with Et2O. The aqueous portion was neutralized with concentrated HCl (523 ~1, 7.0 mmol). The precipitate W O 97126250 PCT~US97/00572 was collected, washed with Et2O, and dried in vac~uo to furnish carbo~;ylic acid 23-3 as a tan solid.
TLC ~f = 0.59 (silica, 20:1 :1 CH2C12/MeOH/AcOH) lH NMR (400 MHz, CD30D) o 9.05 (q, J=2H, lH), 8s.95 (d, J=4H, lH), 8.77 (dd, J=2Hz, 8Hz, lH), 7.67 (~1, J=4H, 1~), 7.53 (d, J=4Hz, lH~, 3.22 (~[, J=~sHz, 2H), 2.46 (t, J=7Hz, 2H), 2.03 (m, 2H).
N~ ~ O
~OEt 4-(1,8-Naphthyridin-4-yl)butanoyl-N-(cyclopropyl)glycine ethyl ester (~3-4) A solution of acid 23-3 (400 mg, 1.~4 mmol), amine 21-3 (331 mg, 1.84 mmol), BOP reagent (979 mg, 2.21 mmol), NMM (1.03 ml, 7.36 mmol) and DMF (20 ml) was stirred at ambient temperature for 20 h. The solution was diluted with ethyl acetate and then washed with sat. NaHCO3, br~ne, dried (Mg~04) and concentrated. Flash chrom~atography (silica, 10:1 EtOAc/sat. NH3-EtOH) furnished ester 23-4 (600 mg) as an orange solid.
TLC Rf = 0.15 (silica, 10:1 EtOAc/sat. NH3-EtOH) lH NMR (300 MHz, CDC13) ~ 9.12 (m, lH), 9.03 (d, J=4Hz, lH), ~s.62 (dd, J-=2Hz, 8Hz, lH), 7.53 (q, J=4Hz, lH), 7.38 (d, J=4Hz, IH), 4.20 (q, J=7Hz, 2H), 4.13 (s, 2H), 3.19 (t, J=8Hz, 2H), 2.79 (m, lH), 2.70 (m, 2H), 2.13 (m, 2H?, 1.29 (t, J=~SHz, 3H), 0.85 (m, 2H), 0.74 (m, 2H).
W O 97/26250 PCTrUS97/00572 - 16~s -HN/ > ~7 o N~/ ~ OEt o ?3-5 4-(1,2,3,4-Tetrahydro- 1,~-naphthyridin-~-~l)butanoyl-N-(cyclo-propyl)~lycine ethyl e~ter (23-5) A mixture of ester 23-4 (600 mg, 1.75 mmol), 10% Pd/C
5 (300 mg) and EtOH (30 ml) was stirred under hydrogen atmosphere ( I
atm) at ambient temperature for 20 h. The catalyst was removed by filtration through celite and then the filtrate was concentrated. Flash chromatography (~silica, 50%/EtOAc/sat. NH3-EtOH) gave ester 23-5 as a colorless oil.
10 TLC Rf = 0.25 (silica, 50:1 EtOAc/sat. NH3-EtOH) H NMR (400 MHz, CD30D) ~ 7.58 (d, J=6Hz, lH), 6.4~ ~d, J=6Hz, IH), 4.15 (q, J=7Hz, 2H), 4.0~ (s, 2H), 3.36 (t, J=5Hz, 2H), 2.~¢6 (m, lH), 2.75 (t, J=6Hz, 2H), 2.6~ (t, J=7Hz, 2H), 2.60 (t, J=~Hz, 2H), 1.90 (m, 4H), 1.25 (t, J=7Hz, 3H), 0.g7 (m, 2H), 0.7~s (m, 2H).~5 r~
HN~ ~ ~7 ~
N~/~ N~J~~H
4-(1,2,3,4-Tetrahydro- 1,~s-napthyridin-5-yl)butanoyl-N-(cyclo-propyl)~lycine (23-6) A solution of ester 23-5 (200 mg, 0.5774 mmole), IN
20 NaOH (600 !11, 0.600 mmole) and CH3OH was stirred at ambient temper~ture for 1.5 h. The ~olution was concentrated. The residue was dissolved in lN HCl (600 ,ul) and then the solution was concentrated.
The residue was dissolved in CHC13, filtered and concentrated to give the carboxylic acid 23-6 (110 mg) as a white solid.
W O 97/26Z50 PCT~US97/00572 TLC ]Rf = 0.14 (~;ilica, 10:1 :1 CH2C12/MeOH/AcOH) IH NMR (300 MHz, CD30D) o 7.56 (d, J=6Hz, 1 H), 6.64 (d, J=6Hz, lH), 3.92 (s, 2H), 3.41 (t, J-6Hz, 2H), 2.g9 (m, IH), 2.gl (t, J=6Hz, 2H), 2.71 (m, 4H), 1.~ (m, 4H), 0.~2 (m, 4H).
s HN/~ ~ o 1 1 N~ N~JlNH~co2Et 4-(1 ,2',3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)~lycyl-3(S)-ethynvl-~-alanine ethyl e~ter (23-7) To a stirred solution of acid 23-6 (40 mg, 0.1256 mmol), amin~ 21-6 (33 mg, 0.18~4 rnrnol), NMM (70 ~11, 0.5024 mmol) and CH3CN (1 ml) was added BOP reagent (61 mg, 0.1382 mmol). After 20 h at arnbient temperature, the solution wa.s diluted with ethyl acetate and then washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 40:1:1 CH2C12/MeOH/AcOH) gave the ester 23-7 as a colorless oil.
TLC Rf = 0.23 (silica, 40:1:1 CH2C12/MeOH/AcOH) H NMR (300 MHz, CD30D) o 7.5g (d, J=6Hz, IH), 6.66 (d, J=6Hz, lH), S.01 (m, lH), 4.13 (q, J=7Hz, 2H), 4.02 (s, 2H), 3.42 (t, J=6Hz, 2H), 2.72 (m, 10H), 1.95 (m, 4H), 1.24 (t, J=7Hz, 3H), 0.P~5 (m, 2H), 20 0.7g (m, 2H).
W 097/262~0 PCT~US97/00572 HN/~ ~77 o l l TFA~ N~JlNH~co2H
4-(1,2,3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl~lycyl-3(S)-ethynyl-,B-alanine (23-8) A solution of ester 23-7 (32 mg, 0.0725 mmol), lN NaOH
S (100 ~1) and CH30H (500 ml) was stirred a~ ambient temperature for 1.0 h. The solution was concentrated. The residue was dis~olved in 1 N
HCI (100 ,ul) and then concentrated. Preparative HPLC purification (Cl~S, H20/CH3CN/TFA) provided acid 23-~s as a TFA salt.
TLC Rf = 0.50 (silica, 10:1:1 EtOH/NH40H/H20) lH NMR (300 MHz, CD30D) ~ 8.43 (d, J=9Hz, IH), 7.58 (d, J=7Hz, lH), 6.76 (d, J=7Hz, lH), 4.99 (m, lH), 4.03 (d, J-3Hz, 2H), 3.46 (t, 3=5Hz, 2H), 2.72 (m, lOH), 1.95 (m, 4H), 0.~7 (m, 2H), 0.79 (m, 2H).
-W ~ 97~Z6250 PCTrUS97/00572 SCHE~ME 24 HCI'~H2N ~ OC2H5 ~ \BOC-Gly ~,N \ BOC-HN ~ ~OC2H5 20-1 ~
N~ ~,N~J~ N~CO2Et 2f ~N~ Z
wo 97126250 PCTAUS97/00572 C02Et N~ N~
l~thyl N-pyridin-4-vlnipecotate (22-1~
Ethyl (~) nipecotate (7.0 g, 44.53 mmol) was reacted with 4-chlorpyridine hydrochloride (6.67 g, 44.53 mmol) as described for 5 21-1 t:o give the title compound as a yellow ~olid.
lH NMR (300 MHz, CDC13) ~ ~.22 (d, J=6.~Hz, 2H), 6.68 (d, J=6.8Hz, 2H), 4.18 (4, J=7.0Hz, 2H), 3.85 (m, lH), 3.72 (m, lH), 3.21 (m, lH), 3.10 (m, lH), 2.60 (m, lH), 2.08 (m, lH), l.gl (m, 2H), 1.60 (m, lH), 1.13 (t, J=7.0Hz, 3H).
~ ~<
N~ N~
N-Pyridin-4-ylnipecotic acid ~22-2) P~epared from 22-1 (764 mg, 3.25 mmol) in a manner similar to that described f~or 21-2.
lH NMR (300 MHz, DMSO-d6) ~ 8.13 (d, J=6.8Hz, 2H), 6.74 (d, J=6.8Hz, 2H), 4.08 (d, lH), 3.78 (m, lH), 2.92 (m, 2H), 2.10 (m, lH), 1.95 (m, lH), 1.71 (m, lH), 1.42 (m, 2H).
W O 97/26250 PCT~US97100572 N3 2f N~,CO2 Bu tert-Butyl N-pyridin-4-ylnipecotyl-N-cyclopropyl~lycine (22-3) Prepared from 22-2 (320 mg, 1.51 mmol) and 21 -3 (258 mg, l.Sl mrnol) in a manner similar to that described for 21-4.
lH NMR (300 MHz, CDC13) ~ 8.12 (d, 6.8Hz, 2H), 6.62 (d, J=6.8Hz, 2H), 3.94 (s, 2H), 3.85 (m, lH), 3.12 (m, lH), 3.08 (m, lH), 251 (m, 2H), 1.95 (m, lH), 1.85 (m, 2H), 1.58 (m, 2H), 1.42 (s, 9H), 0.47 (m, 2H), 0.38 (m, 2H).
N~ N
~HCI
o N-Pyridin-4-ylnipecotyl-N-cyclopropyl~lycine hydrochloride (22-4) Ester 22-3 (250 mg, 0.70 mmol) was suspended in EtOAc (25 mL), cooled to 0~ and treated with HCI gas for 15 min. The resulting solution was stir~ed for 3.5 h then evaporated to give 22-4 as a white solid.
1H NMR (300 MHz, DMSO-d6) ~ 8 18 (d, J=6.8Hz, 2H), 7.1~ (d, J=6.8Hz, 2H), 4.24 (d, J=12.8Hz, 2H), 3.95 (m, 2H), 3.21 (m, IH), 1.94 20 (m, lH), 1.85 (m, lH), 1.72 (m, lH), 1.53 (m, lH), 0.87 (m, 2H), 0.75 (m, 2H).
~ N~ N~ CO~Et Ethy~ N-pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~B-alanine (22-5) Prepared from 22-4 (195 mg, 0.60 mmol) in a manner 5 similar to that described ~or 21-7.
lH NMR (300 MHz, CD30D) ~ 8.49 (d, J=6.~Hz, IH), ~s.17 (d, J=6.~Hz, 2H), 7.21 (d, J=6.8Hz, 2H), 5.15 (m, lH), 4.26 (m, IH), 4.21 (d, lH), 4.08 (q, 2H), 3.82 (m, lH), 3.5-3.3 (m, 3H), 2.95 (m, lH), 2.76 (m, lH), 2.71 (m, 2H), 2.1~ (m, lH), 1.95 (m, lH), 1.~1 (m, lH), 1.72 (m, lH), 1.21 (t~ 3H), O.g7 (m, 2H).
21' ~ N~-- 2 N-Pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-alanine (22-6~
Prepared from 22-4 (20 mg, 0.04 mmol) in a manner 15 simil.~r to that described for 21-8.
FAB mass spectrum m/z = 399 (M + 1).
lH NMR (300 MHz, CD30D) ~ 8.16 (d, J=6Hz, 2H), 6.91 (d, J=6.gHz, 2H), 5.05 (m, lH), 4.26 (d, Hz, lH), 4.21 (d, lH), 3.~s2 (m, lh), 3.5-3.3 (m, 3H), 2.95 (m, lH), 2.76 (m, lH), 2.71 (m, 2H), 2.15 (m, IH), 1.95 (m, lH), l.~sl (m, lH), 1.72 (m, lH), 1.21 (t, lH), 0.87 (m, 2H).
W O 97/262~0 PCTrUS97/00572 CH3 NaN(TMS)2 ~j~ TH F, -78~C N>=~
N N Br/ CO2CH3 N~/\--CO2CH3 1 N NaOH
EtO H
N~9 HCl-HN~C02Et ~==<
N ~ CO2H
BOP, NMM, DMF ~y N~ f ~OEt10 /O Pd/C
W O 97/262~,0 PCTAUS97/00572 - 1~5 -SCHEME 23 (CONT'D) HN/~ ~ 0 1N NaOH
OEt Z3 ~ ~
IJ ~~
HN/ > ~7 0 HCI-H2N~--C~2Et N~ N~J~ BOP, NMM
2q,-6 HN~> ~ H 1. 1N NaOH
N = ~N JINH~CO2Et EtOH
O 2. Prep HPLC
H~ N7~1 H'~~co H
W 097/26250 PCTrUS97/00572 N~
N~--c02CH3 Methyl 4-(l.g-naphthyridin-4-yl)butyrate (23-2) To a stirred solution of naphthyridine 23-1 (Hamada, Y. ~t al., Chenl. Pharm. Bull. Soc., 1971, 19(9), 1857-1862), (2.2 g, 15.2 S rnmol) and THF (200 ml) at -7~~C was added NaN(TMS)2 (lM/THF, l~s ml, l ~S mmol) dropwi.se over a 20 min period. After 30 minutes at -7~~C, methyl 3-bromopropionate was added in a stream. After 30 min, the reaction was (luenched with 50 ml 10% KHSO4. The mixture was extracted with Et2O. The rem~ining aqueous portion was basified with 10 sat. NaHCO3 and then extracted with EtOAc. The EtOAc portion was washed with brine, dried (MgSO4) and concentrated. Flash chromotography (silica, 2% EtOH/EtOAc) gave the ester 23-2 (1.61 g) as a yellow oil.
TLC Rf = 0.27 (silica, 2% EtOH/EtOAc) lH NMR (400 MHz, CDC13) ~ 9.14 (m, lH), 9.35 (d, J=4Hz, lH), g.50 (d, J=7Hz, lH), 7.52 (~1, J=4Hz, IH), 7.33 (d, J=4Hz, lH), 3.71 (s, 3H), 3.14 (t, J=~Hz, 2H), 2.46 (t, J=7Hz, 2H), 2.09 (m, 2H).
N ~ .
N~CO2H
20 4-(l.g-Naphthyridin-4-yl)butanoic acid (1-3) A solution of ester 23-2 (1.60 g, 6.9 mmol), lN NaOH (7 ml, 7 mmol) and EtOH (20 ml) was stirred at ambient temperature for 1.0 h. The solution was extracted with Et2O. The aqueous portion was neutralized with concentrated HCl (523 ~1, 7.0 mmol). The precipitate W O 97126250 PCT~US97/00572 was collected, washed with Et2O, and dried in vac~uo to furnish carbo~;ylic acid 23-3 as a tan solid.
TLC ~f = 0.59 (silica, 20:1 :1 CH2C12/MeOH/AcOH) lH NMR (400 MHz, CD30D) o 9.05 (q, J=2H, lH), 8s.95 (d, J=4H, lH), 8.77 (dd, J=2Hz, 8Hz, lH), 7.67 (~1, J=4H, 1~), 7.53 (d, J=4Hz, lH~, 3.22 (~[, J=~sHz, 2H), 2.46 (t, J=7Hz, 2H), 2.03 (m, 2H).
N~ ~ O
~OEt 4-(1,8-Naphthyridin-4-yl)butanoyl-N-(cyclopropyl)glycine ethyl ester (~3-4) A solution of acid 23-3 (400 mg, 1.~4 mmol), amine 21-3 (331 mg, 1.84 mmol), BOP reagent (979 mg, 2.21 mmol), NMM (1.03 ml, 7.36 mmol) and DMF (20 ml) was stirred at ambient temperature for 20 h. The solution was diluted with ethyl acetate and then washed with sat. NaHCO3, br~ne, dried (Mg~04) and concentrated. Flash chrom~atography (silica, 10:1 EtOAc/sat. NH3-EtOH) furnished ester 23-4 (600 mg) as an orange solid.
TLC Rf = 0.15 (silica, 10:1 EtOAc/sat. NH3-EtOH) lH NMR (300 MHz, CDC13) ~ 9.12 (m, lH), 9.03 (d, J=4Hz, lH), ~s.62 (dd, J-=2Hz, 8Hz, lH), 7.53 (q, J=4Hz, lH), 7.38 (d, J=4Hz, IH), 4.20 (q, J=7Hz, 2H), 4.13 (s, 2H), 3.19 (t, J=8Hz, 2H), 2.79 (m, lH), 2.70 (m, 2H), 2.13 (m, 2H?, 1.29 (t, J=~SHz, 3H), 0.85 (m, 2H), 0.74 (m, 2H).
W O 97/26250 PCTrUS97/00572 - 16~s -HN/ > ~7 o N~/ ~ OEt o ?3-5 4-(1,2,3,4-Tetrahydro- 1,~-naphthyridin-~-~l)butanoyl-N-(cyclo-propyl)~lycine ethyl e~ter (23-5) A mixture of ester 23-4 (600 mg, 1.75 mmol), 10% Pd/C
5 (300 mg) and EtOH (30 ml) was stirred under hydrogen atmosphere ( I
atm) at ambient temperature for 20 h. The catalyst was removed by filtration through celite and then the filtrate was concentrated. Flash chromatography (~silica, 50%/EtOAc/sat. NH3-EtOH) gave ester 23-5 as a colorless oil.
10 TLC Rf = 0.25 (silica, 50:1 EtOAc/sat. NH3-EtOH) H NMR (400 MHz, CD30D) ~ 7.58 (d, J=6Hz, lH), 6.4~ ~d, J=6Hz, IH), 4.15 (q, J=7Hz, 2H), 4.0~ (s, 2H), 3.36 (t, J=5Hz, 2H), 2.~¢6 (m, lH), 2.75 (t, J=6Hz, 2H), 2.6~ (t, J=7Hz, 2H), 2.60 (t, J=~Hz, 2H), 1.90 (m, 4H), 1.25 (t, J=7Hz, 3H), 0.g7 (m, 2H), 0.7~s (m, 2H).~5 r~
HN~ ~ ~7 ~
N~/~ N~J~~H
4-(1,2,3,4-Tetrahydro- 1,~s-napthyridin-5-yl)butanoyl-N-(cyclo-propyl)~lycine (23-6) A solution of ester 23-5 (200 mg, 0.5774 mmole), IN
20 NaOH (600 !11, 0.600 mmole) and CH3OH was stirred at ambient temper~ture for 1.5 h. The ~olution was concentrated. The residue was dissolved in lN HCl (600 ,ul) and then the solution was concentrated.
The residue was dissolved in CHC13, filtered and concentrated to give the carboxylic acid 23-6 (110 mg) as a white solid.
W O 97/26Z50 PCT~US97/00572 TLC ]Rf = 0.14 (~;ilica, 10:1 :1 CH2C12/MeOH/AcOH) IH NMR (300 MHz, CD30D) o 7.56 (d, J=6Hz, 1 H), 6.64 (d, J=6Hz, lH), 3.92 (s, 2H), 3.41 (t, J-6Hz, 2H), 2.g9 (m, IH), 2.gl (t, J=6Hz, 2H), 2.71 (m, 4H), 1.~ (m, 4H), 0.~2 (m, 4H).
s HN/~ ~ o 1 1 N~ N~JlNH~co2Et 4-(1 ,2',3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)~lycyl-3(S)-ethynvl-~-alanine ethyl e~ter (23-7) To a stirred solution of acid 23-6 (40 mg, 0.1256 mmol), amin~ 21-6 (33 mg, 0.18~4 rnrnol), NMM (70 ~11, 0.5024 mmol) and CH3CN (1 ml) was added BOP reagent (61 mg, 0.1382 mmol). After 20 h at arnbient temperature, the solution wa.s diluted with ethyl acetate and then washed with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, 40:1:1 CH2C12/MeOH/AcOH) gave the ester 23-7 as a colorless oil.
TLC Rf = 0.23 (silica, 40:1:1 CH2C12/MeOH/AcOH) H NMR (300 MHz, CD30D) o 7.5g (d, J=6Hz, IH), 6.66 (d, J=6Hz, lH), S.01 (m, lH), 4.13 (q, J=7Hz, 2H), 4.02 (s, 2H), 3.42 (t, J=6Hz, 2H), 2.72 (m, 10H), 1.95 (m, 4H), 1.24 (t, J=7Hz, 3H), 0.P~5 (m, 2H), 20 0.7g (m, 2H).
W 097/262~0 PCT~US97/00572 HN/~ ~77 o l l TFA~ N~JlNH~co2H
4-(1,2,3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl~lycyl-3(S)-ethynyl-,B-alanine (23-8) A solution of ester 23-7 (32 mg, 0.0725 mmol), lN NaOH
S (100 ~1) and CH30H (500 ml) was stirred a~ ambient temperature for 1.0 h. The solution was concentrated. The residue was dis~olved in 1 N
HCI (100 ,ul) and then concentrated. Preparative HPLC purification (Cl~S, H20/CH3CN/TFA) provided acid 23-~s as a TFA salt.
TLC Rf = 0.50 (silica, 10:1:1 EtOH/NH40H/H20) lH NMR (300 MHz, CD30D) ~ 8.43 (d, J=9Hz, IH), 7.58 (d, J=7Hz, lH), 6.76 (d, J=7Hz, lH), 4.99 (m, lH), 4.03 (d, J-3Hz, 2H), 3.46 (t, 3=5Hz, 2H), 2.72 (m, lOH), 1.95 (m, 4H), 0.~7 (m, 2H), 0.79 (m, 2H).
-W ~ 97~Z6250 PCTrUS97/00572 SCHE~ME 24 HCI'~H2N ~ OC2H5 ~ \BOC-Gly ~,N \ BOC-HN ~ ~OC2H5 20-1 ~
24-1a ~HCi, EtOAc O H
BOC ~ ~ ~ OH + H2N ~ ~OC2H5 H O ~ O
~N 2 H
NMM, CH3C~OBT, EDC
o BOC~ ~~,D N ~~f N ~OC2H5 HCi/dioxane H H ~ ~b ~
~4-3 ~ N
W O 97/26250 PCTrUS97/00572 SCHEME 24 (CONT'D) H2N ~ NH ~ ~~C2Hs O ~ O
2 HC I ~ N
~( 24-5 EtOH, Et3N
~N J~ N~ NH ~ OC2H5 H H O
24-6 l~, N
Pd/C, H2 ~MeOH, NH3 ~N J~ N ~ NH ~~ OCH3 H H O O
24-7 ~ N
W 09712625~ PCTAUS97/00572 ~CHEME 24 (CONTrD) Nr N H ~n' OCH3 H H O
24-7 ~ N
HgO, S
EtOH, reflux N H--1~ OCH36N HCI
H H O
~,N
~N N ~ N OH
H H O
~N
W O 97/26250 PCT~US97/00572 Preparation of 3- { 2-[;5-(1 H-Benzoimidazol-2-yl-amino)-pentanoyl-aminol-acetylamino~-3(S)-pyridin-3-yl- propionic acid (24-9) 3-t-Butoxycarbonylaminoacetylamino-3(S)-pyridin-3-yl-propionic acid ethyl ester bis hydrochloride (24- 1 a) A stirred solution of BOC-Gly (645 mg, 3.7 mmol), NMM
(452 uL, 4.0 mmol), and ~tOAc (35 mL) at 0~C was treated with isobutyl chloroformate (534 uL, 4.0 mmol). After 20 min 20-1 (1.0 g, 3.7 mmol) and NMM (1.2 mL, 11 mmol) were added ~ollowed by removal of the cooling bath. After 20 hr, the reaction mixture was wa.shed with H2O, sat. NaHCO3, and brine, dried (MgSO4), and concentrated. Flash chromatography (silica, EtOAc to 5%
MeOH/EtOAc) gave 24-1 as a colorless oil.
TLC: Rf = 0.31 (20% MeOH/EtOAc), lH NMR (300MHz, CDCL3) ~ ~s. 5g (bs, lH), 8.51~m, lH), 7.62 (m, 1 H), 7.49 (m, 1 H), 5.4~ ~m, 1 H), 4.13 (m, 1 H), 4.08 (q, J=7Hz, 2H), 3.~S3 (m, 2H), 2.90 (m, 2H), 1.43 (s, 9H), 1.13 (t, J=7Hz, 3H).
3-Aminoacetylamino-3(S)-pyridin-3-yl-propionic acid ethyl ester bis-hydrochloride (24-2) HCI gas was passed through a solution of 24-la (0.~4 g, 2.4 mmol) in EtOAc (24 mL) at 0~C for 15 min and the reaction mixture stirred ~or an additional 15 min. The reaction mixture was concentrated and the residue triturated with ether to give 24-2 as awhite solid.
TLC: Rf - 0.29 (10: 1: 1 ethanol/H2O/NH4OH).
3-[2-(5-t-Butoxycarbonylaminopentanoylamino)acetylamino~ -3 (S)-pyridin-3-yl-pro~ionic acid ethyl ester (24-3) A CH3CN ~olution (20 mL) of 24-1 (71.7 mg, 0.33mmol), 24-2 (97 mg, 0.30 mmol), HO~3T (50.5 mg, 0.33mmol), EDC (63.3 mg, 0.33 mmol) and NMM (132 ml, 1.2 mmol) was stirred under ambient conditions for 18 hr. The reaction solution was concentrated to a yellow gum which was partitioned between EtOAc and sat. NaC~03 W O 97126250 PCT~US97J00572 solution. The EtOAc layer was washed with H20, brine, dried(MgSO4) and concentrated to provide 24-3 as a colorless gum.
TLC: Rf = 0.41 (50% CH2C12/acetone), lH N]\~R (300MHz, CD~L3) ~ ~S.56(bs, lH), 8.51(m, lH), 7.62(m, lH), 7.49(m, lH), 5.43(m, lH), 4.08(q, J=7Hz, 2H), 3.94(m, 2H), 3.12(m, 2H), ''.90(m, 2H), 2.2~(m, 2H), 1.64(m, 4H), 1.43(s, 9H), 1.13(t, J=7Hz, 3H).
3-[2-(5-Aminopentanoylamino)-acetylamino] -3(S)-pyridin-3-yl-propionic acid ethyl ester dihydrochloride (24-4) A 4M HCI/dioxane solution(10 mL) of 24-3 (101 mg, 0.24 mmol) was stirred under ambient conditions for 18 hr. The solution was concentrated to provide 24-4 as a pale yellow gum which was used in the next step without further purification.
lH N]\~R (300 MHz, CD30D) ~ 8.93(bs, lH), 8.79(m, lH), 8.69(m, lH), ~,.10(m, lH), 5.48(m, lH), 4.14(q, J=7Hz, 2H), 3.gg(m, 2H), 3.07(m, 2H), 2.~9(m, 2H), 2.33(m, 2H), 1.68(m, 4H), 1.23(t, J=7Hz, 3H).
3 -(2- { 5- [3 -(2-Nitrophenyl)-thioureido] -pentanoylamino ~ -acetyl-amino)-3(5)-pvridin-3-yl-propionic acid ethyl ester ~24-6) An ethanol solution(20 mL) of 24-5 (40 mg, 0.224 mmol) and 24-4 (95 mg, 0.224 mmol) was refluxed for 2 hr and concentrated to a yellow gum which was purified by flash chromatography (80%
EtOAc/EtOH-NH3) to provide 24-6 as a yellow gum.
TLC~: Rf = 0.41 (80% EtOAc/EtOH-NH3), lH N]\IR (300 MHz, CD3OD) ~ 8.54(m, IH), 8.42(m, lH), ~.03(m, 2H), 7.P~3(m, IH), 7.63(m, lH), 7.41(m, lH), 7.32(m, lH), 5.39(m, lH), 4.09(~}, J=7 Hz, 2H), 3.86(s, 2H~, 3.61(m, 2H), 2.91(m, 2H), 2.33(m, 2H), 1.69(m, 4H), 1.16(t, J=7Hz, 3H) .
W O 97/26250 PCTrUS97/00572 3-(2- ~ 5-[3-(2-Aminophenyl)-thio~areido]-pentanoylamino ~ -acetylamino)-3(S)-pyridin-3-yl-propionic acid methyl e~ter (24-7) 10% Pd/C (50 mg) and 24-6 (103 mg, 0.194 mmol) were added to methanol saturated with ammonia and the mixture hydrogenated at 1 atm. for lg hr. The reaction wa~s filtered and concentrated to provide 24-7 as a pale yellow gum which was u.sed in the next step without further purification.
lH NMR (300MHz, CD3OD) o 8.53(m, lH), 8.41(m, lH), 7.~S4(m, lH), 7.40(m, lH), 7.07(m, lH), 6.97(m, lH), 6.~2(m, lH), 6.67(m, lH), 10 5.38(m, lH), 3.~5(m, 2H), 3.62(s, 3H). 3.54(m, 2H), 2.93(m, 2H), 2.30(m, 2H), 1.60(m, 4H).
3- { 2-[5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino}-acetyl-amino)-3(S)-pyridin-3-yl-propionic acid methyl e~ter (24-8) An ethanol mixture(20 ml) of 24-7 (89 mg, 0.18 mmol), mercuric oxide (78.8 mg, 0.36 mmol) and sulfur (1.8 mg, 0.056 mmol) wa,s refluxed i~or 2 hr. After cooling, the mixture was filtered and the filtrate concentrated to a semi-solid which was purified by flash chromatography (20 % MeOH/CH2C12) to provide 24-8 as a ,~iolid.
20 TLC: E~f = 0.13 (20% MeOH/CH2C12), lH NMR (300 MHz, CD30D) ~ 8.52(m, lH), 8.41(m, lH), 7.~s1(m, lH), 7.3~(M, lH), 7.23(m, 2H), 7.06(m, 2H), 5.38(m, IH), 3.Ps5(s, 2H), 3.62(,~, 3H), 3.37(m, 2H), 2.95(m, 2H), 2.33(m, 2H), 1.71(m, 4H).
BOC ~ ~ ~ OH + H2N ~ ~OC2H5 H O ~ O
~N 2 H
NMM, CH3C~OBT, EDC
o BOC~ ~~,D N ~~f N ~OC2H5 HCi/dioxane H H ~ ~b ~
~4-3 ~ N
W O 97/26250 PCTrUS97/00572 SCHEME 24 (CONT'D) H2N ~ NH ~ ~~C2Hs O ~ O
2 HC I ~ N
~( 24-5 EtOH, Et3N
~N J~ N~ NH ~ OC2H5 H H O
24-6 l~, N
Pd/C, H2 ~MeOH, NH3 ~N J~ N ~ NH ~~ OCH3 H H O O
24-7 ~ N
W 09712625~ PCTAUS97/00572 ~CHEME 24 (CONTrD) Nr N H ~n' OCH3 H H O
24-7 ~ N
HgO, S
EtOH, reflux N H--1~ OCH36N HCI
H H O
~,N
~N N ~ N OH
H H O
~N
W O 97/26250 PCT~US97/00572 Preparation of 3- { 2-[;5-(1 H-Benzoimidazol-2-yl-amino)-pentanoyl-aminol-acetylamino~-3(S)-pyridin-3-yl- propionic acid (24-9) 3-t-Butoxycarbonylaminoacetylamino-3(S)-pyridin-3-yl-propionic acid ethyl ester bis hydrochloride (24- 1 a) A stirred solution of BOC-Gly (645 mg, 3.7 mmol), NMM
(452 uL, 4.0 mmol), and ~tOAc (35 mL) at 0~C was treated with isobutyl chloroformate (534 uL, 4.0 mmol). After 20 min 20-1 (1.0 g, 3.7 mmol) and NMM (1.2 mL, 11 mmol) were added ~ollowed by removal of the cooling bath. After 20 hr, the reaction mixture was wa.shed with H2O, sat. NaHCO3, and brine, dried (MgSO4), and concentrated. Flash chromatography (silica, EtOAc to 5%
MeOH/EtOAc) gave 24-1 as a colorless oil.
TLC: Rf = 0.31 (20% MeOH/EtOAc), lH NMR (300MHz, CDCL3) ~ ~s. 5g (bs, lH), 8.51~m, lH), 7.62 (m, 1 H), 7.49 (m, 1 H), 5.4~ ~m, 1 H), 4.13 (m, 1 H), 4.08 (q, J=7Hz, 2H), 3.~S3 (m, 2H), 2.90 (m, 2H), 1.43 (s, 9H), 1.13 (t, J=7Hz, 3H).
3-Aminoacetylamino-3(S)-pyridin-3-yl-propionic acid ethyl ester bis-hydrochloride (24-2) HCI gas was passed through a solution of 24-la (0.~4 g, 2.4 mmol) in EtOAc (24 mL) at 0~C for 15 min and the reaction mixture stirred ~or an additional 15 min. The reaction mixture was concentrated and the residue triturated with ether to give 24-2 as awhite solid.
TLC: Rf - 0.29 (10: 1: 1 ethanol/H2O/NH4OH).
3-[2-(5-t-Butoxycarbonylaminopentanoylamino)acetylamino~ -3 (S)-pyridin-3-yl-pro~ionic acid ethyl ester (24-3) A CH3CN ~olution (20 mL) of 24-1 (71.7 mg, 0.33mmol), 24-2 (97 mg, 0.30 mmol), HO~3T (50.5 mg, 0.33mmol), EDC (63.3 mg, 0.33 mmol) and NMM (132 ml, 1.2 mmol) was stirred under ambient conditions for 18 hr. The reaction solution was concentrated to a yellow gum which was partitioned between EtOAc and sat. NaC~03 W O 97126250 PCT~US97J00572 solution. The EtOAc layer was washed with H20, brine, dried(MgSO4) and concentrated to provide 24-3 as a colorless gum.
TLC: Rf = 0.41 (50% CH2C12/acetone), lH N]\~R (300MHz, CD~L3) ~ ~S.56(bs, lH), 8.51(m, lH), 7.62(m, lH), 7.49(m, lH), 5.43(m, lH), 4.08(q, J=7Hz, 2H), 3.94(m, 2H), 3.12(m, 2H), ''.90(m, 2H), 2.2~(m, 2H), 1.64(m, 4H), 1.43(s, 9H), 1.13(t, J=7Hz, 3H).
3-[2-(5-Aminopentanoylamino)-acetylamino] -3(S)-pyridin-3-yl-propionic acid ethyl ester dihydrochloride (24-4) A 4M HCI/dioxane solution(10 mL) of 24-3 (101 mg, 0.24 mmol) was stirred under ambient conditions for 18 hr. The solution was concentrated to provide 24-4 as a pale yellow gum which was used in the next step without further purification.
lH N]\~R (300 MHz, CD30D) ~ 8.93(bs, lH), 8.79(m, lH), 8.69(m, lH), ~,.10(m, lH), 5.48(m, lH), 4.14(q, J=7Hz, 2H), 3.gg(m, 2H), 3.07(m, 2H), 2.~9(m, 2H), 2.33(m, 2H), 1.68(m, 4H), 1.23(t, J=7Hz, 3H).
3 -(2- { 5- [3 -(2-Nitrophenyl)-thioureido] -pentanoylamino ~ -acetyl-amino)-3(5)-pvridin-3-yl-propionic acid ethyl ester ~24-6) An ethanol solution(20 mL) of 24-5 (40 mg, 0.224 mmol) and 24-4 (95 mg, 0.224 mmol) was refluxed for 2 hr and concentrated to a yellow gum which was purified by flash chromatography (80%
EtOAc/EtOH-NH3) to provide 24-6 as a yellow gum.
TLC~: Rf = 0.41 (80% EtOAc/EtOH-NH3), lH N]\IR (300 MHz, CD3OD) ~ 8.54(m, IH), 8.42(m, lH), ~.03(m, 2H), 7.P~3(m, IH), 7.63(m, lH), 7.41(m, lH), 7.32(m, lH), 5.39(m, lH), 4.09(~}, J=7 Hz, 2H), 3.86(s, 2H~, 3.61(m, 2H), 2.91(m, 2H), 2.33(m, 2H), 1.69(m, 4H), 1.16(t, J=7Hz, 3H) .
W O 97/26250 PCTrUS97/00572 3-(2- ~ 5-[3-(2-Aminophenyl)-thio~areido]-pentanoylamino ~ -acetylamino)-3(S)-pyridin-3-yl-propionic acid methyl e~ter (24-7) 10% Pd/C (50 mg) and 24-6 (103 mg, 0.194 mmol) were added to methanol saturated with ammonia and the mixture hydrogenated at 1 atm. for lg hr. The reaction wa~s filtered and concentrated to provide 24-7 as a pale yellow gum which was u.sed in the next step without further purification.
lH NMR (300MHz, CD3OD) o 8.53(m, lH), 8.41(m, lH), 7.~S4(m, lH), 7.40(m, lH), 7.07(m, lH), 6.97(m, lH), 6.~2(m, lH), 6.67(m, lH), 10 5.38(m, lH), 3.~5(m, 2H), 3.62(s, 3H). 3.54(m, 2H), 2.93(m, 2H), 2.30(m, 2H), 1.60(m, 4H).
3- { 2-[5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino}-acetyl-amino)-3(S)-pyridin-3-yl-propionic acid methyl e~ter (24-8) An ethanol mixture(20 ml) of 24-7 (89 mg, 0.18 mmol), mercuric oxide (78.8 mg, 0.36 mmol) and sulfur (1.8 mg, 0.056 mmol) wa,s refluxed i~or 2 hr. After cooling, the mixture was filtered and the filtrate concentrated to a semi-solid which was purified by flash chromatography (20 % MeOH/CH2C12) to provide 24-8 as a ,~iolid.
20 TLC: E~f = 0.13 (20% MeOH/CH2C12), lH NMR (300 MHz, CD30D) ~ 8.52(m, lH), 8.41(m, lH), 7.~s1(m, lH), 7.3~(M, lH), 7.23(m, 2H), 7.06(m, 2H), 5.38(m, IH), 3.Ps5(s, 2H), 3.62(,~, 3H), 3.37(m, 2H), 2.95(m, 2H), 2.33(m, 2H), 1.71(m, 4H).
25 3- { 2-[5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino]-acetyl-amino~-3(S)-pyridin-3-yl-propionic acid (24-9) A 6N HCI solution (5 ml) of 24-8 (33 mg, 0.073 mmol) wa~ ~tirred under ambient condition~ for 18 hr. The reaction wa~
concentrated to give a viscous gum which was purified by prep HPLC
30 (Delta-Pak CI 8, gradient elution over 40 min., 5-50% CH3CN/H20-0.1% TFA) to give 24-9.
lH NMR (300MHz, CD30D) ~ 8.78(m, lH), 8.65(m, lH), ~¢.40(m, IH), 7.g6(m, lH), 7.34(m, 2H), 7.27(m, 2H), 5.40~m, IH), 3.87(m, 2H), 3.42(m, 2H), 2.98(m, 2H), 2.34(M, 2H), 1.74(m, 4H).
W O 97/262S0 PCTrUS97/00572 .
t Tablet Preparation Tablets cont~ining 25.0, 50.0, and 100.0 mg., respectively, of the following active compounds are prepared as illustrated below:
4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl -3(R)-(2-phenethyl)-~-alanine;
4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol-3-yl)ethyl]-~3-alanine; and 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol-3-yl)ethyl]-~-alanine.
TABLE FOR DOSES CONTAIN~G
F~ROM 25-1 00MG OF THE ACTIVE COMPOUND
Amount-m~
Active Compound 25.0 50.0 100.0 Microcrystalline cellulose 37.25 100.0 200.0 Modiiïed food corn starch 37.25 4.25 8.5 Magnesium stearate 0.50 0.75 1.5 All of the active compound, cellulose, and a portion of the corn ,ctarch are mixed and granulated to 10% corn starch paste. The re.sulting granulation is sieved, dried and blended with the remainder of the corn starch and the magnesium stearate. The resulting granulation is then compressed into tablets containing 25.0, 50.0, and 100.0 mg, respectively, of active ingredient per tablet.
W O 97126250PCTrUS97/00572 - 17~ -Intravenous formulations S An intravenous dosage form of the above-indicated active compound i,s prepared as follows:
Active Compound 0.5-lO.Omg Sodium Citrate 5-50mg Citric Acid 1-15mg Sodium Chloride l-~mg Wate~ for Injection (USP) 4.. s. to 1 L
Utilizing the above quantities, the active compound is 10 dissolved at room temperature in a previously prepared solution of sodium chloride, citric acid, and sodium citrate in Water for Injection (IJSP, see page 1636 of United State,s Pharmacopeia/National Formulary for 1995, published by United States Pharmacopeial Convention, Inc., Rockville, Maryland, copyright 1994.
1~
Intravenous formulation A pharmaceutical composition was prepared at room temperature using 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~B-alanine t-butyl ester, a citrate buffer, and sodium chloride, to obtain a concentration of 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-alanine t-butyl ester of 0.25 mg/ml.
gOO grams of water wa.s introduced into a standard pharmaceutical mixing vessel. 0.25 grams of the ester was dissolved in the water. 2.7 grams sodium citrate and 0.16 grams citric acid were added to obtain a fini.~hed citrate concentration of 10 mM. ~ grams of sodium chloride was added. 200 grams of water was then aclded to W 097126250 PCTrUS97/00572 achieve the desired final concentration.s of ingredients. The resulting aque~us formulation had the following concentrations:
In~redient Amount 4-(2-1~minothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,B-alanine t-butyl ester 0.25 mg/ml citrate buffer 10 mM
sodiwn chloride ~g mg/ml The finished concentrated formulation is stored in ~
standard USP Type I borosilicate glass container at 30-40 degrees C.
Prior to compound ~lmini~tration, the concentrated follnulation is 15 diluted in a 4:1 ratio resulting in a finished concentration of 0.05 mg/ml and transfered to an in~usion bag.
Therapeutic Treatment Compounds of the invention may be ~lministered to 20 patients where inhibition of hllm~n or m~mm~lian platelet aggregation or adhesion is desired.
Compounds of the invention are useful in inhibiting platelet aggregation and thus, they may find utility in surgery on peripheral arteries (arterial grafts, carotid endaterectomy~ and in cardiovascular 25 surgery where manipulation of arteries and organs, and/or the interation of pla-telets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelet~s may form thrombi and thromboemboli. Compounds of the invention may be administered to these surgical patients to prevent the formation of thrombi and 30 thromboemboli.
Compounds of the invention are also effective inhibitors of osteoclast cellular adhesion, and can be administered to inhibit bone resorption. The dosage regimen utilizing the compounds of the present invenlion for this purpose is selected in accordance with a variety of W O 97/262S0 PCT~US97/00572 - 1~0-factors. including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of admini,stration; the renal and hepatic function of the patient; and the particu}ar compound or salt thereof employed. An ordinarily skilled S physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the condition.
Oral dosages of the compounds, when used to prevent osteoclast cellular adhesion, will range between about 0.01 mg per kg of 10 body weight per day ~mg/kg/day) to about 100 mg/kg/day and preferably 0.01-50 mg/kg/day and more preferably 0.01-20 mg/kg/day, e.g. 0.1 mg/kg/day, 1.0 mg/kg/day, 5.() mg/kg/day, or 10 mg/kg/day.
Advantageously, compounds of the present invention may be administered in divided doses of two, three, or four time.s daily.
15 Intravenously, the most preferred doses will range from about l to about 10 mg/kg/minute during a con,stant rate infusion. Furthermore, preferred compounds for the present invention can be administered in intranasal form via topical use of suitable intrana,sal vehicles, or via transdermal routes, using tho,se form,s of transderrnal skin patche,s well 20 known to tho.se of ordinary skill in that art. To be administered in the forrn of a transdermal delivery system, the dosage administration will, or course, be continuous rather that intermittent throughout the dosage regime.
25 ETB ~SSAY
Duong et al., J. Bone Min~. Res., 8:S 378, describe a system for expressing the human integrin oCV~3. It has been suggested that the integrin is involved in the attachment of osteoclasts to bone matrix, ~since antibodies against the integrin, or RGD-containing 3~ molecule,s, such as echistatin (European Publication 322 451), can effectively block bone resorption.
W 097126250 PCT~US97/00572 1~1 Reaction Mixture:
1. 175 ,ul TBS buffer (50 mM Tris-HCI pH 7.2, 150 mM
NaC~, 1% BSA, 1 mM CaC12~ 1 mM MgC12).
2. 25 ~I cell extract (di~ute with 100 mM octylglucoside buffer to give 2000 cpm/25 ,ul).
3. 125I-echi~tatin (25 ~11/50,000 cpm) (see EP 382 451).
4. 25 ,ul buffer (total binding) or unlabeled echistatin (non-specific binding).
The reaction mixture was then incubated for 1 h at room temp. The unbound and the bound OCv~3 were separated by filtration using a Skatron C~ell Harvester. The filters (prewet in 1.5% poly-ethyleneimine for 10 mins) were then washed with the wa~h buffer (50 mM Tris HCl, lmM CaC12/MgC12, pH 7.2). The filter was then 15 counted in a gamma counter.
The following compounds were tested and shown to bind to the human integrin ocvl33.
Compound EIB
2-3 < 2000 nM
3-8 < 1000 nM
5-9 < 1000 nM
19-10 < 1000 nM
20-3 < 1000 nM
24-9 < 1000 nM
concentrated to give a viscous gum which was purified by prep HPLC
30 (Delta-Pak CI 8, gradient elution over 40 min., 5-50% CH3CN/H20-0.1% TFA) to give 24-9.
lH NMR (300MHz, CD30D) ~ 8.78(m, lH), 8.65(m, lH), ~¢.40(m, IH), 7.g6(m, lH), 7.34(m, 2H), 7.27(m, 2H), 5.40~m, IH), 3.87(m, 2H), 3.42(m, 2H), 2.98(m, 2H), 2.34(M, 2H), 1.74(m, 4H).
W O 97/262S0 PCTrUS97/00572 .
t Tablet Preparation Tablets cont~ining 25.0, 50.0, and 100.0 mg., respectively, of the following active compounds are prepared as illustrated below:
4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl -3(R)-(2-phenethyl)-~-alanine;
4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol-3-yl)ethyl]-~3-alanine; and 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol-3-yl)ethyl]-~-alanine.
TABLE FOR DOSES CONTAIN~G
F~ROM 25-1 00MG OF THE ACTIVE COMPOUND
Amount-m~
Active Compound 25.0 50.0 100.0 Microcrystalline cellulose 37.25 100.0 200.0 Modiiïed food corn starch 37.25 4.25 8.5 Magnesium stearate 0.50 0.75 1.5 All of the active compound, cellulose, and a portion of the corn ,ctarch are mixed and granulated to 10% corn starch paste. The re.sulting granulation is sieved, dried and blended with the remainder of the corn starch and the magnesium stearate. The resulting granulation is then compressed into tablets containing 25.0, 50.0, and 100.0 mg, respectively, of active ingredient per tablet.
W O 97126250PCTrUS97/00572 - 17~ -Intravenous formulations S An intravenous dosage form of the above-indicated active compound i,s prepared as follows:
Active Compound 0.5-lO.Omg Sodium Citrate 5-50mg Citric Acid 1-15mg Sodium Chloride l-~mg Wate~ for Injection (USP) 4.. s. to 1 L
Utilizing the above quantities, the active compound is 10 dissolved at room temperature in a previously prepared solution of sodium chloride, citric acid, and sodium citrate in Water for Injection (IJSP, see page 1636 of United State,s Pharmacopeia/National Formulary for 1995, published by United States Pharmacopeial Convention, Inc., Rockville, Maryland, copyright 1994.
1~
Intravenous formulation A pharmaceutical composition was prepared at room temperature using 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~B-alanine t-butyl ester, a citrate buffer, and sodium chloride, to obtain a concentration of 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-alanine t-butyl ester of 0.25 mg/ml.
gOO grams of water wa.s introduced into a standard pharmaceutical mixing vessel. 0.25 grams of the ester was dissolved in the water. 2.7 grams sodium citrate and 0.16 grams citric acid were added to obtain a fini.~hed citrate concentration of 10 mM. ~ grams of sodium chloride was added. 200 grams of water was then aclded to W 097126250 PCTrUS97/00572 achieve the desired final concentration.s of ingredients. The resulting aque~us formulation had the following concentrations:
In~redient Amount 4-(2-1~minothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,B-alanine t-butyl ester 0.25 mg/ml citrate buffer 10 mM
sodiwn chloride ~g mg/ml The finished concentrated formulation is stored in ~
standard USP Type I borosilicate glass container at 30-40 degrees C.
Prior to compound ~lmini~tration, the concentrated follnulation is 15 diluted in a 4:1 ratio resulting in a finished concentration of 0.05 mg/ml and transfered to an in~usion bag.
Therapeutic Treatment Compounds of the invention may be ~lministered to 20 patients where inhibition of hllm~n or m~mm~lian platelet aggregation or adhesion is desired.
Compounds of the invention are useful in inhibiting platelet aggregation and thus, they may find utility in surgery on peripheral arteries (arterial grafts, carotid endaterectomy~ and in cardiovascular 25 surgery where manipulation of arteries and organs, and/or the interation of pla-telets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelet~s may form thrombi and thromboemboli. Compounds of the invention may be administered to these surgical patients to prevent the formation of thrombi and 30 thromboemboli.
Compounds of the invention are also effective inhibitors of osteoclast cellular adhesion, and can be administered to inhibit bone resorption. The dosage regimen utilizing the compounds of the present invenlion for this purpose is selected in accordance with a variety of W O 97/262S0 PCT~US97/00572 - 1~0-factors. including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of admini,stration; the renal and hepatic function of the patient; and the particu}ar compound or salt thereof employed. An ordinarily skilled S physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the condition.
Oral dosages of the compounds, when used to prevent osteoclast cellular adhesion, will range between about 0.01 mg per kg of 10 body weight per day ~mg/kg/day) to about 100 mg/kg/day and preferably 0.01-50 mg/kg/day and more preferably 0.01-20 mg/kg/day, e.g. 0.1 mg/kg/day, 1.0 mg/kg/day, 5.() mg/kg/day, or 10 mg/kg/day.
Advantageously, compounds of the present invention may be administered in divided doses of two, three, or four time.s daily.
15 Intravenously, the most preferred doses will range from about l to about 10 mg/kg/minute during a con,stant rate infusion. Furthermore, preferred compounds for the present invention can be administered in intranasal form via topical use of suitable intrana,sal vehicles, or via transdermal routes, using tho,se form,s of transderrnal skin patche,s well 20 known to tho.se of ordinary skill in that art. To be administered in the forrn of a transdermal delivery system, the dosage administration will, or course, be continuous rather that intermittent throughout the dosage regime.
25 ETB ~SSAY
Duong et al., J. Bone Min~. Res., 8:S 378, describe a system for expressing the human integrin oCV~3. It has been suggested that the integrin is involved in the attachment of osteoclasts to bone matrix, ~since antibodies against the integrin, or RGD-containing 3~ molecule,s, such as echistatin (European Publication 322 451), can effectively block bone resorption.
W 097126250 PCT~US97/00572 1~1 Reaction Mixture:
1. 175 ,ul TBS buffer (50 mM Tris-HCI pH 7.2, 150 mM
NaC~, 1% BSA, 1 mM CaC12~ 1 mM MgC12).
2. 25 ~I cell extract (di~ute with 100 mM octylglucoside buffer to give 2000 cpm/25 ,ul).
3. 125I-echi~tatin (25 ~11/50,000 cpm) (see EP 382 451).
4. 25 ,ul buffer (total binding) or unlabeled echistatin (non-specific binding).
The reaction mixture was then incubated for 1 h at room temp. The unbound and the bound OCv~3 were separated by filtration using a Skatron C~ell Harvester. The filters (prewet in 1.5% poly-ethyleneimine for 10 mins) were then washed with the wa~h buffer (50 mM Tris HCl, lmM CaC12/MgC12, pH 7.2). The filter was then 15 counted in a gamma counter.
The following compounds were tested and shown to bind to the human integrin ocvl33.
Compound EIB
2-3 < 2000 nM
3-8 < 1000 nM
5-9 < 1000 nM
19-10 < 1000 nM
20-3 < 1000 nM
24-9 < 1000 nM
Claims (27)
1. A compound of the formula:
and pharmaceutically acceptable salts thereof, wherein X is a 5- or 6-membered monocyclic aromatic ring system containing 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either unsubstituted or substituted with R1 or R2, or a 9- to 10-membered polycyclic ring system, wherein one or more of the rings is aromatic, containing 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either unsubstituted or substituted with R1 or R2, wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, C1, Br, 1, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl, C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy C1-6 alkyloxy and hydroxy, hydroxy C1-6 alkyl;
Y is -(CH2)0-6-, -C ~ C-, -C = C~ , , , , -O-- SO2NH-, -NHSO2-, , , -S(O)0-2~CH2-, , , , , or , where Z is O, NR8, or S; and R8 is defined as R1 above;
R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, triifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl is defined as a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, halogen, hydroxyl, C1-5alkylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-5 alkyl, C1-6alkyl, either unsubstituted or substituted, with one or more groups selected from halogen, hydroxyl, C1-5 alkylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, or hydroxycarbonyl C1-5 alkyl, provided that the carbon atom to which R3 and R4 are attached bears only one heteroatom, -(CH2)m C~CH, -(CH2)m C~C-C1-6 alkyl, -(CH2)m C~C-C3-7cycloalkyl, -(CH2)m C~C- aryl, -(CH2)m C~C-C1-6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C1-6 alkyl, -(CH2)m CH=CH-C3-7cycloalkyl, -(CH2)m CH=CH aryl, -(CH2)m CH=CH C1-6 alkyl aryl, -(CH2)m SO2C1-6 alkyl, or -(CH2)m SO2C1-6 alkylaryl;
R5 is hydrogen, fluorine, C1-8 alkyl, hydroxyl, hydroxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-6 alkyloxy.
C3-8 cycloalkyl, aryl C1-6 alkyloxy, aryl, aryl C1-6 alkyl, C1-6 alkylcarbonyloxy, amino, amino C1-6 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-6 alkyl, aryl amino, aryl amino C1-6 alkyl, aryl C1-6 alkylamino, aryl C1-6 alkylamino C1-6 alkyl, aryl carbonyloxy, aryl C1-6 alkylcarbonyloxy, C1-6 dialkylamino, C1-6 dialkylamino C1-6 alkyl, C1-6 alkylaminocarbonyloxy, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkyloxycarbonylamino, C1-8 alkyloxycarbonylamino C1-8 alkyl, aryl oxycarbonylamino C1-8 alkyl, aryl C1-8 alkyloxycarbonylamino, aryl C1-8 alkyloxycarbonylamino C1-8 alkyl, C1-8 alkylcarbonylamino, C1-8 alkylcarbonylamino C1-6 alkyl, aryl carbonylamino C1-6 alkyl, aryl C1-6 alkylcarbonylamino, aryl C1-6 alkylcarbonylamino C1-6 alkyl, aminocarbonylamino C1-6 alkyl, C1-8 alkylaminocarbonylamino, C1-8 alkylaminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino C1-6 alkyl, aryl C1-8 alkylaminocarbonylamino, aryl C1-8 alkylaminocarbonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl C1-6alkyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, C1-6 alkylcarbonyl, C1-6 alkylcarbonyl C1-6 alkyl, aryl carbonyl C1-6alkyl, aryl C1-6 alkylcarbonyl, aryl C1-6 alkylcarbonyl C1-6alkyl, C1-6 alkylthiocarbonylamino, C1-6 alkylthiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino C1-6 alkyl, aryl C1-6 alkylthiocarbonylamino, aryl C1-6 alkylthiocarbonylamino C1-6 alkyl, aminocarbonyl C1-6 alkyl, or C1-8 alkylaminocarbonyl, or C1-8 alkylaminocarbonyl C1-6 alkyl, or aryl aminocarbonyl C1-6 alkyl, aryl C1-8 alkylaminocarbonyl, aryl C1-8 alkylaminocarbonyl C1-6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2; and R6 and R7 are independently hydrogen, C1-8 alkyl, aryl, aryl C1-8 alkyl, hydroxy, C1-8 alkyloxy, aryloxy, aryl C1-6 alkyloxy, C1-8 alkylcarbonyloxy C1-4 alkyloxy, aryl C1-8 alkylcarbonyloxy C1-4 alkyloxy, C1-8 alkylaminocarbonylmethyleneoxy, or C1-8 dialkylaminocarbonylmethyleneoxy where m and n are integers 0-6.
and pharmaceutically acceptable salts thereof, wherein X is a 5- or 6-membered monocyclic aromatic ring system containing 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either unsubstituted or substituted with R1 or R2, or a 9- to 10-membered polycyclic ring system, wherein one or more of the rings is aromatic, containing 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either unsubstituted or substituted with R1 or R2, wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, C1, Br, 1, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl, C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy C1-6 alkyloxy and hydroxy, hydroxy C1-6 alkyl;
Y is -(CH2)0-6-, -C ~ C-, -C = C~ , , , , -O-- SO2NH-, -NHSO2-, , , -S(O)0-2~CH2-, , , , , or , where Z is O, NR8, or S; and R8 is defined as R1 above;
R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, triifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl is defined as a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, halogen, hydroxyl, C1-5alkylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-5 alkyl, C1-6alkyl, either unsubstituted or substituted, with one or more groups selected from halogen, hydroxyl, C1-5 alkylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, or hydroxycarbonyl C1-5 alkyl, provided that the carbon atom to which R3 and R4 are attached bears only one heteroatom, -(CH2)m C~CH, -(CH2)m C~C-C1-6 alkyl, -(CH2)m C~C-C3-7cycloalkyl, -(CH2)m C~C- aryl, -(CH2)m C~C-C1-6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C1-6 alkyl, -(CH2)m CH=CH-C3-7cycloalkyl, -(CH2)m CH=CH aryl, -(CH2)m CH=CH C1-6 alkyl aryl, -(CH2)m SO2C1-6 alkyl, or -(CH2)m SO2C1-6 alkylaryl;
R5 is hydrogen, fluorine, C1-8 alkyl, hydroxyl, hydroxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-6 alkyloxy.
C3-8 cycloalkyl, aryl C1-6 alkyloxy, aryl, aryl C1-6 alkyl, C1-6 alkylcarbonyloxy, amino, amino C1-6 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-6 alkyl, aryl amino, aryl amino C1-6 alkyl, aryl C1-6 alkylamino, aryl C1-6 alkylamino C1-6 alkyl, aryl carbonyloxy, aryl C1-6 alkylcarbonyloxy, C1-6 dialkylamino, C1-6 dialkylamino C1-6 alkyl, C1-6 alkylaminocarbonyloxy, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkyloxycarbonylamino, C1-8 alkyloxycarbonylamino C1-8 alkyl, aryl oxycarbonylamino C1-8 alkyl, aryl C1-8 alkyloxycarbonylamino, aryl C1-8 alkyloxycarbonylamino C1-8 alkyl, C1-8 alkylcarbonylamino, C1-8 alkylcarbonylamino C1-6 alkyl, aryl carbonylamino C1-6 alkyl, aryl C1-6 alkylcarbonylamino, aryl C1-6 alkylcarbonylamino C1-6 alkyl, aminocarbonylamino C1-6 alkyl, C1-8 alkylaminocarbonylamino, C1-8 alkylaminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino C1-6 alkyl, aryl C1-8 alkylaminocarbonylamino, aryl C1-8 alkylaminocarbonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl C1-6alkyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, C1-6 alkylcarbonyl, C1-6 alkylcarbonyl C1-6 alkyl, aryl carbonyl C1-6alkyl, aryl C1-6 alkylcarbonyl, aryl C1-6 alkylcarbonyl C1-6alkyl, C1-6 alkylthiocarbonylamino, C1-6 alkylthiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino C1-6 alkyl, aryl C1-6 alkylthiocarbonylamino, aryl C1-6 alkylthiocarbonylamino C1-6 alkyl, aminocarbonyl C1-6 alkyl, or C1-8 alkylaminocarbonyl, or C1-8 alkylaminocarbonyl C1-6 alkyl, or aryl aminocarbonyl C1-6 alkyl, aryl C1-8 alkylaminocarbonyl, aryl C1-8 alkylaminocarbonyl C1-6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2; and R6 and R7 are independently hydrogen, C1-8 alkyl, aryl, aryl C1-8 alkyl, hydroxy, C1-8 alkyloxy, aryloxy, aryl C1-6 alkyloxy, C1-8 alkylcarbonyloxy C1-4 alkyloxy, aryl C1-8 alkylcarbonyloxy C1-4 alkyloxy, C1-8 alkylaminocarbonylmethyleneoxy, or C1-8 dialkylaminocarbonylmethyleneoxy where m and n are integers 0-6.
2. A compound of Claim 1 having the formula:
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , , , , , or , wherein n is 2-4, and n' is 2 or 3, and wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy C1-6 alkyloxy and hydroxy, hydroxy C1-6 alkyl;
R5 is;
hydrogen, fluorine, C1-8 alkyl, hydroxyl, hydroxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-6 alkyloxy, C3-8 cycloalkyl, aryl C1-6 alkyloxy, aryl, aryl C1-6 alkyl, C1-6 alkylcarbonyloxy, amino, C 1-6 alkylamino, amino C1-6 alkyl, C1-6 alkylamino C1-6 alkyl, aryl amino, aryl amino C1-6 alkyl, aryl C1-6 alkylamino, aryl C1-6 alkylamino C1-6 alkyl, aryl carbonyloxy, aryl C1-6 alkylcarbonyloxy, C1-6 dialkylamino, C1-6 dialkylamino C1-6 alkyl, C1-6 alkylaminocarbonyloxy, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkyloxycarbonylamino, C1-8 alkyloxycarbonylamino C1-8 alkyl, aryl C1-8 alkyloxycarbonylamino, aryl oxycarbonylamino, aryl oxycarbonylamino C1-8 alkyl, aryl C1-8 alkyloxycarbonylamino C1-8 alkyl, C1-8 alkylcarbonylamino, C1-8 alkylcarbonylamino C1-6 alkyl, aryl carbonylamino C1-6 alkyl, aryl carbonylamino, aryl C1-6 alkylcarbonylamino, aryl C1-6 alkylcarbonylamino C1-6 alkyl, C1-8 alkylaminocarbonylamino, aminocarbonylamino, aminocarbonylamino C1-6 alkyl, C1-8 alkylaminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino, aryl C1-8 alkylaminocarbonylamino, aryl C1-8 alkylaminocarbonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, aminosulfonylamino, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl, aryl sulfonyl C1-6alkyl, aryl alkylsulfonyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, C1-6 alkylcarbonyl, C1-6 alkylcarbonyl C1-6 alkyl, aryl carbonyl C1-6alkyl, aryl carbonyl, aryl C1-6 alkylcarbonyl, aryl C1-6 alkylcarbonyl C1-6alkyl, C1-6 alkylthiocarbonylamino, C1-6 alkylthiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino, aryl C1-6 alkylthiocarbonylamino, aryl C1-6 alkylthiocarbonylamino C1-6 alkyl, aminocarbonyl C1-6 alkyl, aminocarbonyl, C1-8 alkylaminocarbonyl, C1-8 alkylaminocarbonyl C1-6 alkyl, aryl aminocarbonyl C1-6 alkyl, aryl aminocarbonyl, aryl C1-8 alkylaminocarbonyl, aryl C1-8 alkylaminocarbonyl C1-6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2; and R6 and R7 are independently hydrogen, C1-8 alkyl, aryl C1-8 alkyl, hydroxy, C1-8 alkyloxy, aryl, aryl C1-6 alkyloxy, C1-8 alkylcarbonyloxy C1-4 alkyloxy, aryl C1-8 alkylcarbonyloxy C1-4 alkyloxy, C1-8 alkylaminocarbonylmethyleneoxy, or C1-8 dialkylaminocarbonylmethyleneoxy, where m and n are integers 0-6.
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , , , , , or , wherein n is 2-4, and n' is 2 or 3, and wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy C1-6 alkyloxy and hydroxy, hydroxy C1-6 alkyl;
R5 is;
hydrogen, fluorine, C1-8 alkyl, hydroxyl, hydroxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-6 alkyloxy, C3-8 cycloalkyl, aryl C1-6 alkyloxy, aryl, aryl C1-6 alkyl, C1-6 alkylcarbonyloxy, amino, C 1-6 alkylamino, amino C1-6 alkyl, C1-6 alkylamino C1-6 alkyl, aryl amino, aryl amino C1-6 alkyl, aryl C1-6 alkylamino, aryl C1-6 alkylamino C1-6 alkyl, aryl carbonyloxy, aryl C1-6 alkylcarbonyloxy, C1-6 dialkylamino, C1-6 dialkylamino C1-6 alkyl, C1-6 alkylaminocarbonyloxy, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkyloxycarbonylamino, C1-8 alkyloxycarbonylamino C1-8 alkyl, aryl C1-8 alkyloxycarbonylamino, aryl oxycarbonylamino, aryl oxycarbonylamino C1-8 alkyl, aryl C1-8 alkyloxycarbonylamino C1-8 alkyl, C1-8 alkylcarbonylamino, C1-8 alkylcarbonylamino C1-6 alkyl, aryl carbonylamino C1-6 alkyl, aryl carbonylamino, aryl C1-6 alkylcarbonylamino, aryl C1-6 alkylcarbonylamino C1-6 alkyl, C1-8 alkylaminocarbonylamino, aminocarbonylamino, aminocarbonylamino C1-6 alkyl, C1-8 alkylaminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino, aryl C1-8 alkylaminocarbonylamino, aryl C1-8 alkylaminocarbonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, aminosulfonylamino, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl, aryl sulfonyl C1-6alkyl, aryl alkylsulfonyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, C1-6 alkylcarbonyl, C1-6 alkylcarbonyl C1-6 alkyl, aryl carbonyl C1-6alkyl, aryl carbonyl, aryl C1-6 alkylcarbonyl, aryl C1-6 alkylcarbonyl C1-6alkyl, C1-6 alkylthiocarbonylamino, C1-6 alkylthiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino, aryl C1-6 alkylthiocarbonylamino, aryl C1-6 alkylthiocarbonylamino C1-6 alkyl, aminocarbonyl C1-6 alkyl, aminocarbonyl, C1-8 alkylaminocarbonyl, C1-8 alkylaminocarbonyl C1-6 alkyl, aryl aminocarbonyl C1-6 alkyl, aryl aminocarbonyl, aryl C1-8 alkylaminocarbonyl, aryl C1-8 alkylaminocarbonyl C1-6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2; and R6 and R7 are independently hydrogen, C1-8 alkyl, aryl C1-8 alkyl, hydroxy, C1-8 alkyloxy, aryl, aryl C1-6 alkyloxy, C1-8 alkylcarbonyloxy C1-4 alkyloxy, aryl C1-8 alkylcarbonyloxy C1-4 alkyloxy, C1-8 alkylaminocarbonylmethyleneoxy, or C1-8 dialkylaminocarbonylmethyleneoxy, where m and n are integers 0-6.
3. A compound of Claim 2 having the formula:
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , , , , , , , wherein n' is 2 or 3, and wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy C1-6 alkyloxy and hydroxy, hydroxy C1-6 alkyl;
Y is -(CH2)0-6- , -C ~ C-, -C = C ~ , , , , -O-, - SO2NH - , -NHSO2- , , , , , , or , where Z is O, NR8, or S; and R8 is defined as R1 above;
R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups ,selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl is defined as a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, halogen, hydroxyl, C1 5alkylcarbonylamino, C1-5 alkoxylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-5 alkyl, C1-6alkyl, either unsubstituted or substituted, with one or more groups selected from halogen, hydroxyl, C1-5alkylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, or hydroxycarbonyl C1-5 alkyl, provided that the carbon atom to which R3 and R4 are attached bears only one heteroatom, -(CH2)m C~CH, -(CH2)m C~C-C1-6 alkyl, -(CH2)m C~C-C3-7cycloalkyl, -(CH2)m C~C- aryl, -(CH2)m C~C-C1-6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C1-6 alkyl, -(CH2)m CH=CH-C3-7cycloalkyl, -(CH2)m CH=CH aryl, -(CH2)m CH=CH C1-6 alkyl aryl, -(CH2)mSO2C1-6alkyl, or -(CH2)mSO2C1-6alkylaryl;
R5 is hydrogen, fluorine, C 1-8 alkyl, hydroxyl, hydroxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-6 alkyloxy.
C3-8 cycloalkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyl, C1-6 alkylcarbonyloxy, amino C1-6 alkyl, amino, C1-6 alkylamino, C1-6 alkylamino C1-6 alkyl, aryl amino C1-6 alkyl, aryl amino, aryl C1-6 alkylamino, aryl C1-6 alkylamino C1-6 alkyl, aryl, aryl C1-6 alkylcarbonyloxy, C1-6 dialkylamino, C1-6 dialkylamino C1-6 alkyl, C1-6 alkylaminocarbonyloxy, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkyloxycarbonylamino, C1-8 alkyloxycarbonylamino C1-8 alkyl, aryl oxycarbonylamino C1-8 alkyl, aryl oxycarbonylamino, aryl C1-8 alkyloxycarbonylamino, aryl C1-8 alkyloxycarbonylamino C1-8 alkyl, C1-8 alkylcarbonylamino, C1-8 alkylcarbonylamino C1-6 alkyl, aryl carbonylamino C1-6 alkyl, aryl carbonylamino, aryl C1-6 alkylcarbonylamino, aryl C1-6 alkylcarbonylamino C1-6 alkyl, aminocarbonylamino C1-6 alkyl, aminocarbonylamino, C1-8 alkylaminocarbonylamino, C1-8 alkylaminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino, aryl C1-8 alkylaminocarbonylamino, aryl C1-8 alkylaminocarbonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, aminosulfonylamino, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl C1-6alkyl, aryl sulfonyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, C1-6 alkylcarbonyl, C1-6 alkylcarbonyl C1-6 alkyl, aryl carbonyl C1-6alkyl, aryl carbonyl, aryl C1-6 alkylcarbonyl, aryl C1-6 alkylcarbonyl C1-6alkyl, C1-6 alkylthiocarbonylamino, C1-6 alkylthiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino, aryl C1-6 alkylthiocarbonylamino, aryl C1-6 alkylthiocarbonylamino C1-6 alkyl, aminocarbonyl C1-6 alkyl, aminocarbonyl, C1-8 alkylaminocarbonyl, C1-8 alkylaminocarbonyl C1-6 alkyl, aryl aminocarbonyl C1-6 alkyl, aryl aminocarbonyl, aryl C1-8 alkylaminocarbonyl, or aryl C1-8 alkylaminocarbonyl C1-6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2; and R6 and R7 are independently hydrogen, C1-8 alkyl, aryl, aryl C1-8 alkyl, hydroxy, C1-8 alkyloxy, aryl oxy, aryl C1-6 alkyloxy, C1-8 alkylcarbonyloxy C1-4 alkyloxy, aryl C1-8 alkylcarbonyloxy C1-4 alkyloxy, C1-8 alkylaminocarbonylmethyleneoxy, or C1-8 dialkylaminocarbonylmethyleneoxy, where m and n are integers 0-6.
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , , , , , , , wherein n' is 2 or 3, and wherein R1 and R2 are independently selected from the group consisting of hydrogen, F, Cl, Br, I, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy C1-6 alkyloxy and hydroxy, hydroxy C1-6 alkyl;
Y is -(CH2)0-6- , -C ~ C-, -C = C ~ , , , , -O-, - SO2NH - , -NHSO2- , , , , , , or , where Z is O, NR8, or S; and R8 is defined as R1 above;
R3 and R4 are independently hydrogen, a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups ,selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl is defined as a five or six membered mono or polycyclic aromatic ring system containing 0, 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, halogen, hydroxyl, C1 5alkylcarbonylamino, C1-5 alkoxylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-5 alkyl, C1-6alkyl, either unsubstituted or substituted, with one or more groups selected from halogen, hydroxyl, C1-5alkylcarbonylamino, arylC1-5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1-5 alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, aminoC1-3 alkyl, arylaminocarbonyl, arylC1-5alkylaminocarbonyl, aminocarbonyl-C1-4 alkyl, hydroxycarbonyl, or hydroxycarbonyl C1-5 alkyl, provided that the carbon atom to which R3 and R4 are attached bears only one heteroatom, -(CH2)m C~CH, -(CH2)m C~C-C1-6 alkyl, -(CH2)m C~C-C3-7cycloalkyl, -(CH2)m C~C- aryl, -(CH2)m C~C-C1-6 alkyl aryl, -(CH2)m CH=CH2, -(CH2)m CH=CH C1-6 alkyl, -(CH2)m CH=CH-C3-7cycloalkyl, -(CH2)m CH=CH aryl, -(CH2)m CH=CH C1-6 alkyl aryl, -(CH2)mSO2C1-6alkyl, or -(CH2)mSO2C1-6alkylaryl;
R5 is hydrogen, fluorine, C 1-8 alkyl, hydroxyl, hydroxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-6 alkyloxy.
C3-8 cycloalkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyl, C1-6 alkylcarbonyloxy, amino C1-6 alkyl, amino, C1-6 alkylamino, C1-6 alkylamino C1-6 alkyl, aryl amino C1-6 alkyl, aryl amino, aryl C1-6 alkylamino, aryl C1-6 alkylamino C1-6 alkyl, aryl, aryl C1-6 alkylcarbonyloxy, C1-6 dialkylamino, C1-6 dialkylamino C1-6 alkyl, C1-6 alkylaminocarbonyloxy, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkyloxycarbonylamino, C1-8 alkyloxycarbonylamino C1-8 alkyl, aryl oxycarbonylamino C1-8 alkyl, aryl oxycarbonylamino, aryl C1-8 alkyloxycarbonylamino, aryl C1-8 alkyloxycarbonylamino C1-8 alkyl, C1-8 alkylcarbonylamino, C1-8 alkylcarbonylamino C1-6 alkyl, aryl carbonylamino C1-6 alkyl, aryl carbonylamino, aryl C1-6 alkylcarbonylamino, aryl C1-6 alkylcarbonylamino C1-6 alkyl, aminocarbonylamino C1-6 alkyl, aminocarbonylamino, C1-8 alkylaminocarbonylamino, C1-8 alkylaminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino C1-6 alkyl, aryl aminocarbonylamino, aryl C1-8 alkylaminocarbonylamino, aryl C1-8 alkylaminocarbonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, aminosulfonylamino, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl C1-6alkyl, aryl sulfonyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, C1-6 alkylcarbonyl, C1-6 alkylcarbonyl C1-6 alkyl, aryl carbonyl C1-6alkyl, aryl carbonyl, aryl C1-6 alkylcarbonyl, aryl C1-6 alkylcarbonyl C1-6alkyl, C1-6 alkylthiocarbonylamino, C1-6 alkylthiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino C1-6 alkyl, aryl thiocarbonylamino, aryl C1-6 alkylthiocarbonylamino, aryl C1-6 alkylthiocarbonylamino C1-6 alkyl, aminocarbonyl C1-6 alkyl, aminocarbonyl, C1-8 alkylaminocarbonyl, C1-8 alkylaminocarbonyl C1-6 alkyl, aryl aminocarbonyl C1-6 alkyl, aryl aminocarbonyl, aryl C1-8 alkylaminocarbonyl, or aryl C1-8 alkylaminocarbonyl C1-6 alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2; and R6 and R7 are independently hydrogen, C1-8 alkyl, aryl, aryl C1-8 alkyl, hydroxy, C1-8 alkyloxy, aryl oxy, aryl C1-6 alkyloxy, C1-8 alkylcarbonyloxy C1-4 alkyloxy, aryl C1-8 alkylcarbonyloxy C1-4 alkyloxy, C1-8 alkylaminocarbonylmethyleneoxy, or C1-8 dialkylaminocarbonylmethyleneoxy, where m and n are integers 0-6.
4. A compound of Claim 3 having the formula:
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , , , , , , or wherein R1 and R2 are independently selected from the group consisting of hydrogen or amino, amino C1-8 alkyl;
Y is -(CH2)0-4- , , -O-, or ;
R8 is hydrogen or aryl C0-8 alkyl;
R3 is hydrogen, a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl is defined as a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, C3-8 cycloalkyl, or C1-6alkyl, either unsubstituted or substituted, with C3-8 cycloalkyl;
R4 is hydrogen, -(CH2)n-aryl, wherein n=0-4 and aryl is defined as a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonylC0-5 alkyl, or hydroxycarbonylC1-5 alkoxy, C1-6alkyl, or -(CH2)0-4 C~CH;
R5 is hydrogen, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, aminosulfonylamino, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl C1-6alkyl, aryl sulfonyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2;
R6 is hydrogen, C1-8 alkyl, or aryl, aryl C1-8 alkyl;
m is an integer selected from 0 to 6; and n is an integer selected from 0 to 6.
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , , , , , , or wherein R1 and R2 are independently selected from the group consisting of hydrogen or amino, amino C1-8 alkyl;
Y is -(CH2)0-4- , , -O-, or ;
R8 is hydrogen or aryl C0-8 alkyl;
R3 is hydrogen, a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, -(CH2)n-aryl, wherein n=1-4 and aryl is defined as a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonyl, hydroxycarbonylC1-5 alkyl, or hydroxycarbonylC1-5 alkoxy, C3-8 cycloalkyl, or C1-6alkyl, either unsubstituted or substituted, with C3-8 cycloalkyl;
R4 is hydrogen, -(CH2)n-aryl, wherein n=0-4 and aryl is defined as a six membered monocyclic aromatic ring system, either unsubstituted or substituted, with one or more groups selected from hydroxyl, halogen, cyano, trifluoromethyl, C1-3 alkoxy, C1-5 alkylcarbonyloxy, C1-5 alkoxycarbonyl, C1-5 alkyl, aminoC1-5 alkyl, hydroxycarbonylC0-5 alkyl, or hydroxycarbonylC1-5 alkoxy, C1-6alkyl, or -(CH2)0-4 C~CH;
R5 is hydrogen, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, C1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino C1-6 alkyl, aryl sulfonylamino C1-6 alkyl, aryl sulfonylamino, aryl C1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamino C1-6 alkyl, aminosulfonylamino C1-6 alkyl, aminosulfonylamino, C1-8 alkylaminosulfonylamino, C1-8 alkylaminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino C1-6 alkyl, aryl aminosulfonylamino, aryl C1-8 alkylaminosulfonylamino, aryl C1-8 alkylaminosulfonylamino C1-6 alkyl, C1-6 alkylsulfonyl, C1-6 alkylsulfonyl C1-6alkyl, aryl sulfonyl C1-6alkyl, aryl sulfonyl, aryl C1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1-6alkyl, wherein alkyl groups and aryl groups may be unsubstituted or substituted with one or more substituents selected from R1 and R2;
R6 is hydrogen, C1-8 alkyl, or aryl, aryl C1-8 alkyl;
m is an integer selected from 0 to 6; and n is an integer selected from 0 to 6.
5. A compound of Claim 4 having the formula:
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , ;
, ;
, or Y is -(CH2)0-4- , -O- , , or ;
R3 is hydrogen, methyl, , or ;
R4 is hydrogen, methyl, , , , ~ CH;
R5 is hydrogen, or ;
R6 is hydrogen, methyl, ethyl, or t-butyl;
m is an integer selected from 0 to 6; and n is an integer selected from 0 to 6.
and pharmaceutically acceptable salts thereof, wherein X is , , , , , , ;
, ;
, or Y is -(CH2)0-4- , -O- , , or ;
R3 is hydrogen, methyl, , or ;
R4 is hydrogen, methyl, , , , ~ CH;
R5 is hydrogen, or ;
R6 is hydrogen, methyl, ethyl, or t-butyl;
m is an integer selected from 0 to 6; and n is an integer selected from 0 to 6.
6. A compound of claim 5 selected from the group consisting of 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-.beta.-alanine t-butyl ester, 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-.beta.-alanine, 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-.beta.-alanine methyl ester, 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-.beta.-alanine trifluoroacetate salt, 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-.beta.-alanine ethyl ester, 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-.beta.-alanine trifluoroacetate salt, 4-(2-Bocamino-pyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-yl)ethyl]-.beta.-alanine ethyl ester, 4-(2-Aminopyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-yl)ethyl]-.beta.-alanine, 4-(2-Boc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-.beta.-alanine t-butyl ester, 4-(2-Aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-.beta.-alanine, 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-.beta.-alanine ethyl ester, 4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-.beta.-alanine, 4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-.beta.-alanine ethyl ester, 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phenethyl)-.beta.-alanine ethyl ester hydrochloride, 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-phene thyl)-.beta.-alanine, 4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol-3-yl)ethyl]-.beta.-alanine, 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol-3-yl)ethyl]-.beta.-alanine, 4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-.beta.-alanine ethyl ester, 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-.beta.-alanine ethyl ester, 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-.beta.-alanine, 4-(Pyridin-4-yl)butanoyl-N-(2-phenylethyl)glycyl-3(R)-(2-phenethyl)-.beta.-alanine ethyl ester, 4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-.beta.-alanine, 4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-.beta.-alanine benzyl ester, 4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-.beta.-alanine, 4-(2-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-.beta.-alanine, 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-.beta.-alanine ethyl ester, 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-.beta.-alanine, 3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-.beta.-alanine ethyl ester, 3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-phenethyl)-.beta.-alanine, N-{N'-3-(4-t-Butoxycarbonyl-1-piperizinyl)benzoyl)glycyl}-3(R)-methyl-.beta.-alanine benzyl ester, N-[N'-[3-(1-Piperazinyl)benzoyl]glycyl]-3(R)-methyl-.beta.-alanine trifluoroacetic acid salt, N-[N'-[3-(4-t-Butoxycarbonyl-1-piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-.beta.-alanine methyl ester, N-[N'-[3-(1-Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-.beta.-alanine trifluoroacetic acid salt, N-[N'-[3-(4-t-Butoxycarbonyl-1-piperazinyl)benzoyl]-N'-(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-.beta.-alanine methyl ester, N-[N'-[3-(1-Piperazinyl)benzoyl]-N'-(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-.beta.-alanine trifluoroacetic acid salt, 4-(1,2,3,4-Tetrahydro-1,8-naphthyridin-7-yl)butanoyl-glycyl-.beta.-alanine t-butyl ester, 4-(1,2,3,4-Tetrahydro-1,8-naphthyridin-7-yl)butanoyl-glycyl-.beta.-alaninte, 4-(1,2,3,4-Tetrahydro-1,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-pyridin-3-yl-.beta.-alanine ethyl ester, 4-(1,2,3,4-Tetrahydro-1,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-pyridin-3-yl-.beta.-alanine, Ethyl N-pyridin-4-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-.beta.-alanine, N-Pyridin-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-.beta.-alanine, Ethyl N-pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-.beta.-alanine, N-Pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-.beta.-alanine, 4-(1,2,3,4-Tetrahydro-1,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3(S)-ethynyl-.beta.-alanine ethyl ester, 4-(1,2,3,4-Tetrahydro-1,8-naphthyridin-5-yl)butanoyl-N-(cyclo-propyl)glycyl-3(S)-ethynyl-.beta.-alanine, 3-{2-[5-(1H-Benzoimidazol-2-yl-amino)-pentanoylamino]-acetylamino}-3(S)-pyridin-3-yl- propionic acid, and and pharmaceutically acceptable salts thereof.
7. A compound of Claim 1 for use in inhibiting the binding of fibrinogen to blood platelets, inhibiting the aggregation of blood platelets, treating thrombus formation or embolus formation, or preventing thrombus or embolus formation in a mammal.
8. A composition for inhibiting the binding of fibrinogen to blood platelets in a mammal, comprising an antifibrinogenic binding effective amount of a compound of Claim 1 and a pharmaceutically acceptable carrier.
9. A composition for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising an antifibrinogenic binding effective amount of a compound of Claim 1 and a pharmaceutically acceptable carrier.
10. A composition for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising an antifibrinogenic binding effective amount of a compound of Claim 1 in combination with a thrombolytic agent and a pharmaceutically acceptable carrier.
11. A composition for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising an antifibrinogenic binding effective amount of a compound of Claim 1 in combination with an anticoagulant agent and pharmaceutically acceptable carrier.
12. A method for inhibiting the binding of fibrinogen to blood platelets in a mammal, comprising administering to the mammal a composition of Claim 8.
13. A method for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising administering to the mammal a composition of Claim 9.
14. A method for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising administering to the mammal a composition of Claim 10.
15. A method for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising administering to the mammal a composition of Claim 11.
16. A composition for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising an antifibrinogenic binding effective amount of a compound of Claim 1 in combination with two or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent and a pharmaceutically acceptable carrier.
17. A method for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising administering to the mammal the composition of Claim 16.
18. A compound of Claim 5 for use in inhibiting the binding of fibrinogen to blood platelets, inhibiting the aggregation of blood platelets, treating thrombus formation or embolus formation, or preventing thrombus or embolus formation in a mammal.
19. A composition for inhibiting the binding of fibrinogen to blood platelets, in a mammal, comprising an antifibrinogenic binding effective amount of a compound of Claim 6 and a pharmaceutically acceptable carrier.
20. A composition for inhibiting the aggregation of blood platelets, in a mammal, by blocking fibrinogen from acting at its receptor site, comprising an antifibrinogenic binding effective amount of a compound of Claim 6 and a pharmaceutically acceptable carrier.
21. A method for inhibiting the binding of fibrinogen to blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising administering to the mammal a composition of Claim 19.
22. A method for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising administering to the mammal a composition of Claim 20.
23. A composition for inhibiting osteoclast cellular adhesion to mammalin bone surfaces comprising a compound of Claim 1 and a pharmaceutically acceptable carrier.
24. A composition for inhibiting the solubilization of mammalian bone minerals by osteoclast cells in a mammal comprising a compound of Claim 1 and a pharmaceutically acceptable carrier.
25. A method for inhibiting osteoclast cellular adhesion to mammalin bone surfaces comprising treating the mammal with a pharmacologically effective amount of a composition of Claim 23.
26. A method for inhibiting the solubilization of mammalian bone minerals by osteoclast cells in a mammal comprising treating the mammal with a pharmacologically effective amount of a composition of Claim 24.
27. The use of a compound of Claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for inhibiting the aggregation of blood platelets, preventing platelet thrombosis, preventing thromboembolism or preventing reocclusion, in a mammal.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US996596P | 1996-01-16 | 1996-01-16 | |
| US60/009,965 | 1996-01-16 | ||
| GB9603373.3 | 1996-02-16 | ||
| GBGB9603373.3A GB9603373D0 (en) | 1996-02-16 | 1996-02-16 | Integrin receptor antagonists |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2242877A1 true CA2242877A1 (en) | 1997-07-24 |
Family
ID=26308745
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002242877A Abandoned CA2242877A1 (en) | 1996-01-16 | 1997-01-13 | Integrin receptor antagonists |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0880511A4 (en) |
| JP (1) | JP2000516197A (en) |
| AU (1) | AU713676B2 (en) |
| CA (1) | CA2242877A1 (en) |
| WO (1) | WO1997026250A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5981546A (en) * | 1996-08-29 | 1999-11-09 | Merck & Co., Inc. | Integrin antagonists |
| US6211184B1 (en) | 1996-08-29 | 2001-04-03 | Merck & Co., Inc. | Integrin antagonists |
| US5919792A (en) * | 1996-10-30 | 1999-07-06 | Merck & Co., Inc. | Integrin antagonists |
| US5952341A (en) * | 1996-10-30 | 1999-09-14 | Merck & Co., Inc. | Integrin antagonists |
| US6017925A (en) * | 1997-01-17 | 2000-01-25 | Merck & Co., Inc. | Integrin antagonists |
| US6294549B1 (en) | 1997-07-23 | 2001-09-25 | Merck & Co., Inc. | Method for eliciting an αvβ5 or dual αvβ3/αvβ5 antagonizing effect |
| GB2327609A (en) * | 1997-07-23 | 1999-02-03 | Merck & Co Inc | A method for eliciting an avß5 or dual avß3/avß5 antagonizing effect |
| US6048861A (en) | 1997-12-17 | 2000-04-11 | Merck & Co., Inc. | Integrin receptor antagonists |
| EE200000362A (en) * | 1997-12-17 | 2001-12-17 | Merck & Co., Inc. | Integrin receptor antagonists, a pharmaceutical composition and a process for its preparation |
| US6001855A (en) * | 1998-01-02 | 1999-12-14 | Hoffman-La Roche Inc. | Thiazole derivatives |
| EP1054871A2 (en) | 1998-04-01 | 2000-11-29 | Du Pont Pharmaceuticals Company | Pyrimidines and triazines as integrin antagonists |
| EP1074543B1 (en) | 1998-04-09 | 2012-01-25 | Meiji Seika Pharma Co., Ltd. | Aminopiperidine derivatives as integrin alpha v beta 3 antagonists |
| DE19831710A1 (en) * | 1998-07-15 | 2000-01-20 | Merck Patent Gmbh | New diacyl-hydrazine derivatives, are integrin inhibitors useful for treating e.g. thrombosis, cardiac infarction, tumors, osteoporosis, inflammation or infection |
| CN1146541C (en) | 1999-02-20 | 2004-04-21 | 默克专利股份有限公司 | Beta-alanine derivatives |
| EP1181278B1 (en) * | 1999-05-14 | 2004-09-29 | Bristol-Myers Squibb Pharma Research Labs, Inc. | Cyclic amine derivatives and their uses |
| DK1187592T3 (en) | 1999-06-02 | 2007-11-05 | Merck & Co Inc | Alpha v integrin receptor antagonists |
| AU6316900A (en) | 1999-08-05 | 2001-03-05 | Meiji Seika Kaisha Ltd. | Omega-amino-alpha-hydroxycarboxylic acid derivatives having integrin alphavbeta3antagonism |
| EP1743885B1 (en) * | 2000-05-31 | 2009-11-11 | Santen Pharmaceutical Co., Ltd. | 1-[(adamantyl)alkyl]-3-[(pyridinyl)alkyl]urea compounds as TNF-.alpha inhibitors useful for autoimmune diseases |
| DE10041423A1 (en) * | 2000-08-23 | 2002-03-07 | Merck Patent Gmbh | biphenyl |
| JP5021152B2 (en) * | 2001-10-22 | 2012-09-05 | ザ スクリプス リサーチ インスティチュート | Integrin targeting compounds |
| US7317019B2 (en) | 2003-08-21 | 2008-01-08 | Bristol Myers Squibb Co. | N-alkylated diaminopropane derivatives as modulators of chemokine receptor activity |
| PL3929196T3 (en) | 2013-09-24 | 2023-12-11 | Fujifilm Corporation | Pharmaceutical composition of a nitrogen-containing compound or salt thereof, or metal complex thereof |
| EP3760202A1 (en) | 2017-02-28 | 2021-01-06 | Morphic Therapeutic, Inc. | Inhibitors of (alpha-v)(beta-6) integrin |
| RU2769702C2 (en) | 2017-02-28 | 2022-04-05 | Морфик Терапьютик, Инк. | Avb6 integrin inhibitors |
| AU2019327475A1 (en) | 2018-08-29 | 2021-03-11 | Morphic Therapeutic, Inc. | INHIBITING αV β6 INTEGRIN |
Family Cites Families (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3553227A (en) * | 1968-08-05 | 1971-01-05 | Parke Davis & Co | S-2-((2-(2-thiazolylcarbamoyl)ethyl)amino)ethyl thiosulfate, salts thereof, and methods for their production |
| US4684722A (en) * | 1986-01-06 | 1987-08-04 | E. R. Squibb & Sons, Inc. | Monosulfactams |
| CA2008116C (en) * | 1989-02-23 | 2001-11-20 | Thomas Weller | Glycine derivatives |
| US5332726A (en) * | 1989-09-29 | 1994-07-26 | Rhone-Poulenc Rorer Pharmaceuticals Inc. | Antithrombotic peptides and pseudopeptides |
| US5051405A (en) * | 1989-10-10 | 1991-09-24 | Rhone-Poulenc Rorer Pharmaceuticals Inc. | Anti-thrombotic peptides and pseudopeptides |
| HUT68769A (en) * | 1991-05-07 | 1995-07-28 | Merck & Co Inc | FIBRINOGéN RECEPTOR ANTAGONIST COMPOUNDS AND PHARMACEUTICAL COMPOSITIONS COMPRISING THEM AS EFFECTIVE SUBSTANCE |
| DE4212304A1 (en) * | 1992-04-13 | 1993-10-14 | Cassella Ag | Aspartic acid derivatives, their preparation and use |
| FR2692899B1 (en) * | 1992-06-30 | 1994-08-19 | Adir | New peptide derivatives active in cell adhesion processes, their preparation process and the pharmaceutical compositions containing them. |
| US5726192A (en) * | 1992-12-29 | 1998-03-10 | Smithkline Beecham Corporation | Platelet aggregation inhibiting compounds |
| DE4309867A1 (en) * | 1993-03-26 | 1994-09-29 | Cassella Ag | New urea derivatives, their production and use |
| CA2155307A1 (en) * | 1993-03-29 | 1994-10-13 | Michael Garth Wayne | Heterocyclic compounds as platelet aggregation inhibitors |
| AU7862794A (en) * | 1993-10-19 | 1995-05-08 | Sumitomo Pharmaceuticals Company, Limited | 2,3-diaminopropionic acid derivative |
| CN1209060A (en) * | 1995-12-29 | 1999-02-24 | 史密丝克莱恩比彻姆公司 | Vitronectin receptor antagonists |
-
1997
- 1997-01-13 JP JP09526121A patent/JP2000516197A/en active Pending
- 1997-01-13 AU AU16990/97A patent/AU713676B2/en not_active Ceased
- 1997-01-13 WO PCT/US1997/000572 patent/WO1997026250A1/en not_active Application Discontinuation
- 1997-01-13 CA CA002242877A patent/CA2242877A1/en not_active Abandoned
- 1997-01-13 EP EP97902931A patent/EP0880511A4/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| EP0880511A1 (en) | 1998-12-02 |
| AU713676B2 (en) | 1999-12-09 |
| AU1699097A (en) | 1997-08-11 |
| WO1997026250A1 (en) | 1997-07-24 |
| EP0880511A4 (en) | 1999-06-16 |
| JP2000516197A (en) | 2000-12-05 |
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