Thrombin inhibitors based on the amino acid sequence of hirudin

• US 6,060,451 A
• Filed: 03/20/1995
• Issued: 05/09/2000
• Est. Priority Date: 06/15/1990
• Status: Expired due to Fees

• Current Assignee: National Research Council Canada
• Original Assignee: National Research Council Canada
• Inventors: Steinmetzer, Torsten, Konishi, Yasuo, Ni, Feng, DiMaio, John
• Primary Examiner(s): Tsang, Cecilia J.
• Assistant Examiner(s): Delacroix-Muirheid, C.
• Application Number:
  US08/406,142
• Time in Patent Office: 1,877 Days
• Field of Search: 514/13-17, 530/325, 530/329, 530/332
• US Class Current: 514/14.7
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: C07K 14/815 from leeches, e.g. hirudin, eglin

Abstract

A thrombin inhibitor comprising a first bulky hydrophobic portion interacting with the catalytic site of thrombin responsible for proteolysis and a second portion at least maintaining the hydrophobic and acidic character of amino acids 55 to 60 of native hirudin at the C-terminal non-catalytic region of N-acetyl-hirudin45-65. Between the first and second portions is a divalent linker moiety having a chain length of at least 10 carbon atoms. Connecting the first bulky hydrophobic portion and the linker is a peptidomimetic bond. Preferably, the bulky hydrophobic portion comprises at least one amino acid of D-configuration. The compounds are useful in the treatment of thrombotic disorders.

Claims

1. A thrombin inhibitor of formula (I)

View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 40, 47, 48, 49, 50, 51, 52, 53)

2. A thrombin inhibitor according to claim 1, wherein Y is carbonyl.

3. A thrombin inhibitor according to claim 2, wherein Z is a divalent linker moiety composed of a carbon chain that is interrupted by one or more O, S, or NH, and can be substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, alkoxyalkyl, aryl, and aralkyl groups that can in turn be substituted by amide, hydroxy, imidazol, carboxy, or halogen groups, the terminal atom of the chain being a carbon atom that is part of a carbonyl group that forms a peptide linkage with said acidic portion A and with the proviso that the initial atom of the chain is not a nitrogen atom forming a peptide bond with said Y.

4. A thrombin inhibitor according to claim 3, wherein Z is

5. A thrombin inhibitor according to claim 4, wherein E is the native hirudin^49-54 sequence or analogs thereof.

6. A thrombin inhibitor according to claim 4, wherein E is selected from the group consisting of -Gln-Ser-His-Asn-Asp-Gly-, -Gly-Ser-His-Asn-Asp-Gly-, -[5-aminovaleryl]_1-2, and (Gly)₄.

7. A thrombin inhibitor according to claim 3, wherein Z is (CH₂)_2-4 (CO)-Gln-Ser-His-Asn-Asp-Gly-.

8. A thrombin inhibitor according to claim 7, wherein Z is (CH₂)₂ (CO)-Gln-Ser-His-Asn-Asp-Gly-.

9. A thrombin inhibitor according to claim 7, wherein Z is (CH₂)₃ (CO)-Gln-Ser-His-Asn-Asp-Gly-.

10. A thrombin inhibitor according to claim 7, wherein Z is selected from the group consisting of (CH₂)₄ (CO)-Gln-Ser-His-Asn-Asp-Gly-.

11. A thrombin inhibitor according to claim 4, wherein E is a synthetic spanner of the general formula (III):

space="preserve" listing-type="equation">--[NH--L--CO].sub.m -- (III)

wherein m is an integer ranging from 1 to 4, and L is a hexapeptide, or saturated or unsaturated alkyl chain corresponding to 18 atoms or less.

12. A thrombin inhibitor according to claim 11, wherein L is --(CH_0-2)_n --, wherein n is an integer ranging from 1 to 4.

13. A thrombin inhibitor according to claim 12, wherein L is --(CH_0-2)₄ --.

14. A thrombin inhibitor according to claim 13, wherein L is --CH₂ --CH₂ --CH₂ --CH₂ --.

15. A thrombin inhibitor according to claim 13, wherein L is --CH₂ --CH═

CH--CH₂ --.

16. A thrombin inhibitor according to claim 4, wherein Z is (CH₂)₄ CO--[NH--CH₂ --CH═

CH--CH₂ --CO]_1-3.

17. A thrombin inhibitor according to claim 4, wherein Z is (CH₂)₄ CO--[NH--CH₂ --CH═

CH--CH₂ --CO]₁.

18. A thrombin inhibitor according to claim 4, wherein Z is (CH₂)₄ CO--[NH--CH₂ --CH═

CH--CH₂ --CO]₂.

19. A thrombin inhibitor according to claim 4, wherein Z is (CH₂)₄ CO--[NH--CH₂ --CH═

CH--CH₂ --CO]₃.

20. A thrombin inhibitor according to claim 4, wherein E is a carbon chain interrupted by O, NH, or S, and can be substituted by one or more substituents selected from the group consisting of alkyl, alkoxy, and alkoxyalkyl groups that can be substituted in turn by amide, hydroxy, imidazol, carboxy, or halogen groups, wherein said alkyl, alkoxy, and alkoxyalkyl groups can join the carbon chain at two points to form an aromatic or non-aromatic ring within the carbon chain.

21. A thrombin inhibitor according to claim 20, wherein E is pyridylacetyl-R¹⁵ wherein R¹⁵ is 1-4 amino acids.

22. A thrombin inhibitor according to claim 21, wherein E is 4-pyridylacetyl-(R²⁰)_0-1 -R³⁰ wherein R²⁰, if present, is a linker of formula (III), --[NH--L--CO]_m --, wherein m is 1-6 and R³⁰ is (Gly)_1-4.

23. A thrombin inhibitor according to claim 22, wherein R²⁰ is not present, and R³⁰ is (Gly)_2-4.

24. A thrombin inhibitor according to claim 1 or 2 wherein said AS portion is:

##STR32## wherein X is a hydrophobic group;

B is a residue of a hydrophobic amino acid;

W is H or a branched or straight chained alkyl, aryl, or aralkyl radical;

D is ρ

-phenylmethyl, ρ

-phenylethyl, or a linear carbon chain having 2 to 4 carbon atoms which can be substituted by lower alkyl.

25. A thrombin inhibitor according to claim 24, wherein X is a hydrophobic α

-amino acid in the D-configuration, attached by a peptide linkage to B.

26. A thrombin inhibitor according to claim 25, wherein X is D-Phe, D-4FPhe or D-4ClPhe wherein the α

-amino group is neutralized by acetylation or benzoylation.

27. A thrombin inhibitor according to claim 24, wherein B is a residue of a hydrophobic α

-amino acid of the L-configuration or a cyclic imino acid which can bear one or more alkyl substituents attached to the ring, wherein said substituents may bridge to form a cyclic structure;

D is ρ

-phenylmethyl, ρ

-phenylethyl, ethylene, butylene, or propylene;

W is hydrogen or a lower alkyl, aryl, or aralkyl substituent.

28. A thrombin inhibitor according to claim 27 wherein B contains a piperidine or pyrrolidine ring;

and

W is H or a lower alkyl, aryl, or aralkyl substituent on the 3, 4 or 5 position of the piperidine or pyrrolidine ring of B.

29. A thrombin inhibitor according to claim 1, wherein

X is D-Phe, D-4FPhe or D-4ClPhe;

B is valine, pipecolic acid, or proline;

W is hydrogen or a lower alkyl, aryl, or aralkyl substituent and may be substituted on the 3, 4, or 5 position of the ring when B is proline or pipecolic acid;

D is propylene or ρ

-phenylmethyl.

30. A thrombin inhibitor according to claim 28, wherein

X is D-Phe,

B is Pro, and

D is propylene.

31. A thrombin inhibitor according to claim 1 wherein G, G'"'"', X'"'"', and Q are present, Q¹ is Ile and Q² is Pro.

32. A thrombin inhibitor according to claim 1, wherein G and G'"'"' are independently Asp, Glu, ##STR33## wherein n is 0 to 2,

X'"'"' is L-Phe, L-4FPhe, or L-4ClPhe, Phe, Glu, or Tyr;

Q is proline, pipecolic acid, sarcosine, or glutamic acid;

R is selected from the group consisting of ##STR34## wherein J is H, or OH, substituted at the para position of the phenyl ring;

R'"'"' is hydroxyl radical of the COOH terminus of the preceding amino acid, a group comprising 1 to 2 amino acid, or an amine group having the following formula;

##STR35## wherein R₂ and R₃ are each independently hydrogen, lower alkyl, aryl, or alkoxyalkyl, or may be joined to form a ring of 5-6 members, which ring can optionally contain an additional heteroatom selected from O, S, and NH, or pharmaceutically acceptable salts thereof.

33. A thrombin inhibitor according to claim 32, wherein

G and G'"'"' are independently Asp, or Glu;

X'"'"' is Phe;

Q is proline, pipecolic acid, or Glu;

R is selected from the group consisting of ##STR36## wherein J is H, or OH, substituted at the para position of the phenyl ring;

R'"'"' and R" are hydroxyl radical of the COOH terminus of the preceding amino acid, any amino acid, or an amine group having the following formula;

##STR37## wherein R₂ and R₃ are each independently hydrogen, lower alkyl, aryl, or alkoxyalkyl, or may be joined to form a ring of 5-6 members, which ring can optionally contain an additional heteroatom selected from the group consisting of O, S, and NH, or pharmaceutically acceptable salts thereof.

34. A thrombin inhibitor according to claim 1, wherein

G is Asp,

G'"'"' is Glu,

X'"'"' is Phe,

Q is Glu or Pro,

W'"'"' is H, n-butyl, or methyl, and

R is Leu-R'"'"', ##STR38## wherein J is H, or OH, substituted at the para position of the phenyl ring, and

R'"'"' and R" are a hydroxyl radical of the COOH terminus of the preceding amino acid, any amino acid, or an amine group or pharmaceutically acceptable salts thereof.

35. A thrombin inhibitor according to claim 34, wherein R is Glu-Glu-Tyr-Leu-Gln-OH, or Leu-OH, or Glu-Glu-Tyr-Leu-OH.

36. A thrombin inhibitor according to claim 34, wherein R is Glu-Glu-Tyr-Leu-Gln-OH, or Leu-OH.

40. A compound according to claim 3 selected from the group consisting of:

P79 (SEQ ID NO;

1) (BCH-1709);

Ac(D-Phe)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P102 (SEQ ID NO;

2);

Ac(D-Phe)-Pro-Arg-(CH₂)₃ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P103 (SEQ ID NO;

3);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P109 (SEQ ID NO;

4);

Ac(D-Phe)-Pro-Arg-CH₂ (CO)-GSHNDGDFEEIPEEYLQ-OH;

P183 (SEQ ID NO;

5);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]-DFEPIPL-OH;

P184 (SEQ ID NO;

6) (BCH-1710);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]₂ -DFEPIPL-OH;

P185 (SEQ ID NO;

7);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]₃ -DFEPIPL-OH;

P290 (SEQ ID NO;

8) (BCH-1719);

Ac(D-Phe)-thioPro-Arg-(CH₂)₂ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P311 (SEQ ID NO;

9) (BCH-1721);

Ac(D-Cha)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P314 (SEQ ID NO;

10) (BCH-1726);

Ac(D-Cha)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEPIPEEY-(Cha)-Q-OH;

P536 (SEQ ID NO;

11);

(D-Cha)-Pro-Arg-CH₂ N(COCH₃)CH₂ (CO)-(Gly)₄ -DYEPIPEEY-(Cha)-D-OH;

P574 (SEQ ID NO;

12);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(Gly)₄ -DFEEIPEEYLQ-OH;

P596 (SEQ ID NO;

13) (BCH-2773);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(GLY)₄ -DYEPIPEEACha-(D)Glu-OH;

P597 (SEQ ID NO;

14);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(Gly)₄ -DFEEIPEEYLQ-OH;

P603 (SEQ ID NO;

15);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Abu-Gly-DYEPIPEEA-(Cha)-)D-Glu)-OH;

P604 (SEQ ID NO;

16);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Ava-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P605 (SEQ ID NO;

17);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Aca-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P606 (SEQ ID NO;

18) (BCH-2774);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Aha-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P609 (SEQ ID NO;

19);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(Gly)₄ -DYEPIPEEA-(Cha)-(D-Glu)-OH;

P617 (SEQ ID NO;

20) (BCH-2772);

H-(D-Phe)-Pro-Arg-CH₂ SCH₂ (CO)-(Gly)₄ -DFEEPIEEYLQ-OH;

P618 (SEQ ID NO;

21);

Ac(D-Phe)-Pro-Arg-CH₂ SCH₂ (CO)-(Gly)₄ -DFEEPIPEEYLQ-OH;

P658 (SEQ ID NO;

22);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-NH(CH₂)₂ CO-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P659 (SEQ ID NO;

23);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Gly-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P660 (SEQ ID NO;

24);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-NH(CH₂)₃ CO-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P661 (SEQ ID NO;

25);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-NH(CH₂)₂ CO-DYEPIPEEA-(Cha)-(D-Glu)-OH;

BCH-2408 (SEQ ID NO;

26);

Ac(D-Phe)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEPIPEEY-(Cha)-Q-OH;

BCH-2414 (SEQ ID NO;

27);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]₂ -DEFPIPY-OH;

BCH-2423 (SEQ ID NO;

28);

succinyl(D-Phe)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEEIPEEYLQ-OH;

BCH-2739 (SEQ ID NO;

29);

AC(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DFEPIPL-OH;

BCH-2757 (SEQ ID NO;

30);

alpha-N-(Ac)(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]-NGDFEPIPL-TFA salt;

BCH-2758 (SEQ ID NO;

31);

(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DF-(4-thiahomoGlu)-PIPL-TFA salt;

BCH-2763 (SEQ ID NO;

32);

(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DFEPIPL-TFA salt; and

BCH-2767 (SEQ ID NO;

33);

(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -FEPIPL-TFA salt.

47. A pharmaceutical composition containing a compound according to claim 3 wherein said compound is present in admixture with a pharmaceutically acceptable carrier.

48. A pharmaceutical composition containing a compound according to claim 40 wherein said compound is present in admixture with a pharmaceutically acceptable carrier.

49. A pharmaceutical composition containing a compound according to claim 3 wherein said compound is present in admixture with another therapeutically active agent.

50. A pharmaceutical composition containing a compound according to claim 40 wherein said compound is present in admixture with another therapeutically active agent.

51. A method for the treatment or prevention of vascular diseases of a mammal, including human, comprising the administration of an effective amount of a compound according to claim 3.

52. A method for the treatment or prevention of vascular diseases of a mammal, including human, comprising the administration of an effective amount of a compound according to claim 40.

53. A method for the treatment or prevention of vascular diseases of a mammal, including human, comprising the administration of an effective amount of a composition according to anyone of claims 47 to 52.

37. A peptide derivative having the following formula VI:

##STR39## wherein X is a hydrophobic group;

B is a residue of a hydrophobic amino acid;

W is H, or a branched or straight chain alkyl, aryl or aralkyl radical;

D is a linear carbon chain having 2 to 4 carbon atoms which can be substituted by lower alkyl, or D is a p-phenylmethyl or a p-phenylethyl group;

Y is carbonyl, hydroxymethyl of either D or L configuration or --CH₂ --;

Z is a divalent straight chained link moiety that having a chain length of at least about 10 atoms, in the case where Y is carbonyl or hydroxymethyl, the atom adjacent to the carbon atom of Y may be unsubstituted or mono- or di-fluoro-substituted, and Z can be composed of a carbon chain that is interrupted by one or more O, S, NH, carbonyl, ester or amide groups and can be substituted by one or more substituents selected from alkyl, alkoxy, alkoxyalkyl, aryl and aralkyl groups that can in turn be substituted by amide, hydroxy, imidazol, carboxy or halogen groups, the terminal atom of the chain being a carbon atom that is part of a carbonyl group that forms a peptide linkage with the L-α

-amino acid G.

G and G'"'"' are the same or different and are an L-α

-amino acid having a pk value of about 5 or below;

X'"'"' is a hydrophobic L-α

-amino acid;

Q is a residue of a L-α

-amino acid or a cyclic L-imino acid;

W'"'"' is H, or a branched or straight chain alkyl, aryl or aralkyl radical;

R is a hydrophobic group comprising 1 to 5 amino acids or an alkyl, aryl, or aralkyl radical which can be substituted by a carboxyl or amide function or, ##STR40## substituted at the para position of the phenyl ring;

or R and R'"'"' are the same or different and are an amino acid or an amine group having the following formula;

##STR41## wherein R₂ and R₃ are each independently hydrogen, lower alkyl, aryl or alkoxyalkyl, and may be joined to form a ring of 5-6 members, which ring can optionally contain an additional heteroatom selected from O, S, and NH, and pharmaceutically acceptable salts thereof, or

R'"'"' is a hydroxyl group attached to Leu to form a carboxyl group, or a therapeutically acceptable salt thereof.

38. A peptide according to claim 37, wherein

X is a hydrophobic α

-amino acid in the D-configuration, attached by a peptide linkage to substituent B or a hydrophobic group attached to the nitrogen atom of substituent B;

B is a residue of a hydrophobic α

-amino acid or the L-configuration or a cyclic imino acid, which can bear one or more alkyl substituents attached to the ring, which substituents may bridge to form a cyclic structure;

W is H or a lower alkyl, aryl or aralkyl substituent on the 3, 4, or 5 position of the piperidine or pyrrolidine ring;

D is p-phenylmethyl, p-phenylethyl, ethylene, butylene, or propylene;

Y is carbonyl;

Z has a chain length between 12 and 40 and preferably 20 atoms;

G and G'"'"' are the same or different and are Asp, Glu, ##STR42## wherein n is 1 or 2, X'"'"' is L-phe, L4FPhe or L-4ClPhe;

Q is proline, pipecolic acid, sarcosine or Glu;

W'"'"' is H or a lower alkyl, aryl or aralkyl substituent on the 3, 4 or 5 position of the piperidine or pyrrolidine ring;

W'"'"' is H or a lower alkyl, aryl or aralkyl substituent on the 3, 4 or 5 position of the piperidine or pyrrolidine ring;

##STR43## J is H, OH, ##STR44## substituted in the para position and R'"'"' is a hydroxyl group or ##STR45## wherein R₂ and R₃ are straight chain or branched alkyl chains having 1 to 6 carbon atoms and may be joined to form a ring of 5-6 members.

39. A peptide according to claim 37, wherein

X is D-Phe, D-4FPhe or D-4ClPhe wherein the α

-amino group is neutralized by acetylation or benzoylation, or X is naphthalenesulfonyl, benzenesulfonyl, toluenesulfonyl;

B is Val, pipecolic acid or Pro;

W is hydrogen or a lower alkyl, aryl or aralkyl substituent on the 3,4 or 5 position of the piperidine or pyrrolidine ring;

D is propylene of the ring when B is Pro or pipecolic acid or phenylmethyl;

Y is carbonyl;

Z is ##STR46## wherein n is an integer ranging from 1 to 4,

the native hirudin48-54 sequence, or

a synthetic spanner of the general formula ##STR47## wherein n is an integer ranging from 1 to 4; and

L is a hexapeptide or saturated or unsaturated alkyl chain corresponding to 18 atoms or less of a hexapeptide;

R is Glu-Glu-Tyr-Leu-Gln-OH, Leu-OH or Glu-Glu-Tyr-Leu-R'"'"' wherein R'"'"' is a hydroxyl group or a group having the following formula;

##STR48## wherein R₂ is --CH₃ or phenylethyl, R₃ is H or --CH₃ or R₂ and R₃ may be joined together to form --CH₂ --CH₂ --CH₂ --CH₂ --, --CH₂ --CH₂ --CH₂ --CH₂ --CH₂ -- or --CH₂ --CH₂ --O--CH₂ --CH₂ --.

41. A thrombin inhibitor selected from the group consisting of:

P536 (SEQ ID NO;

11);

(D-Cha)-Pro-Arg-CH₂ N(COCH₃)CH₂ (CO)-(Gly)₄ -DYEPIPEEY-(Cha)-D-OH;

P617 (SEQ ID NO;

20) (BCH-2772);

H-(D-Phe)-Pro-Arg-CH₂ SCH₂ (CO)-(Gly)₄ -DFEEPIEEYLQ-OH; and

P618 (SEQ ID NO;

21);

Ac(D-Phe)-Pro-Arg-CH₂ SCH₂ (CO)-(Gly)₄ -DFEEPIPEEYLQ-OH.

42. A thrombin inhibitor selected from the group consisting of:

P574 (SEQ ID NO;

12);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(Gly)₄ -DFEEIPEEYLQ-OH;

P596 (SEQ ID NO;

13) (BCH-2773);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(GLY)₄ -DYEPIPEEACha-(D)Glu-OH;

P597 (SEQ ID NO;

14);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(Gly)₄ -DFEEIPEEYLQ-OH;

P603 (SEQ ID NO;

15);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Abu-Gly-DYEPIPEEA-(Cha)-)D-Glu)-OH;

P604 (SEQ ID NO;

16);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Ava-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P605 (SEQ ID NO;

17);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Aca-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P606 (SEQ ID NO;

18) (BCH-2774);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Aha-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P609 (SEQ ID NO;

19);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(Gly)₄ -DYEPIPEEA-(Cha)-(D-Glu)-OH;

P658 (SEQ ID NO;

22);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-NH(CH₂)₂ CO-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P659 (SEQ ID NO;

23);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Gly-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH;

P660 (SEQ ID NO;

24);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-NH(CH₂)₃ CO-DYEPIPEEA-(Cha)-(D-Glu)-OH; and

P661 (SEQ ID NO;

25);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-NH(CH₂)₂ CO-DYEPIPEEA-(Cha)-(D-Glu)-OH.

43. A thrombin inhibitor selected from the group consisting of:

P183 (SEQ ID NO;

5);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]-DFEPIPL-OH;

P184 (SEQ ID NO;

6) (BCH-1710);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]₂ -DFEPIPL-OH;

P185 (SEQ ID NO;

7);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]₃ -DFEPIPL-OH;

BCH-2414 (SEQ ID NO;

27);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]₂ -DEFPIPY-OH;

BCH-2739 (SEQ ID NO;

29);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DFEPIPL-OH;

BCH-2757 (SEQ ID NO;

30);

alpha-N-(Ac)(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]-NGDFEPIPL-TFA salt;

BCH-2758 (SEQ ID NO;

31);

(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DF-(4-thiahomoGlu)-PIPL-TFA salt; and

BCH-2763 (SEQ ID NO;

32);

(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DFEPIPL-TFA salt.

44. A thrombin inhibitor selected from the group consisting of:

P79 (SEQ ID NO;

1) (BCH-1709);

Ac(D-Phe)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P103 (SEQ ID NO;

3);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P311 (SEQ ID NO;

9) (BCH-1721);

Ac(D-Cha)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEEIPEEYLQ-OH;

P314 (SEQ ID NO;

10) (BCH-1726);

Ac(D-Cha)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEPIPEEY-(Cha)-Q-OH;

BCH-2408 (SEQ ID NO;

26);

Ac(D-Phe)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEPIPEEY-(Cha)-Q-OH;

BCH-2414 (SEQ ID NO;

27);

Ac(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--CH₂ --CH═

CH--CH₂ --(CO)]₂ -DEFPIPY-OH;

BCH-2423 (SEQ ID NO;

28);

succinyl(D-Phe)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEEIPEEYLQ-OH;

BCH-2763 (SEQ ID NO;

32);

(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DFEPIPL-TFA salt;

P596 (SEQ ID NO;

13) (BCH-2773);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(GLY)₄ -DYEPIPEEACha-(D)Glu-OH; and

P606 (SEQ ID NO;

18) (BCH-2774);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Aha-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH.

45. A thrombin inhibitor selected from the group consisting of:

P314 (SEQ ID NO;

10) (BCH-1726);

Ac(D-Cha)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEPIPEEY-(Cha)-Q-OH;

BCH-2408 (SEQ ID NO;

26);

Ac(D-Phe)-Pro-Arg-(CH₂)₂ (CO)-QSHNDGDFEPIPEEY-(Cha)-Q-OH;

BCH-2763 (SEQ ID NO;

32);

(D-Phe)-Pro-Arg-(CH₂)₄ (CO)--[NH--(CH₂)₄ (CO)]₂ -DFEPIPL-TFA salt;

P596 (SEQ ID NO;

13) (BCH-2773);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-(GLY)₄ -DYEPIPEEACha-(D)Glu-OH; and

P606 (SEQ ID NO;

18) (BCH-2774);

(D-Cha)-Pro-Arg-CH₂ (4-pyridylacetyl)-Aha-Gly-DYEPIPEEA-(Cha)-(D-Glu)-OH.

46. A thrombin inhibitor:

BCH-2763 (SEQ ID NO;

32);

(D)Phe-Pro-Arg-(CH₂)₄ (CO)--[NH(CH₂)₄ CO]₂ -DFEPIPL-TFA salt.