Novel microproteins

• US 5,089,474 A
• Filed: 05/24/1990
• Issued: 02/18/1992
• Est. Priority Date: 05/31/1989
• Status: Expired due to Fees

• Current Assignee: Roussel Uclaf
• Original Assignee: Roussel Uclaf
• Inventors: Castro, Bertrand, Lenguyen, Dung, Previero, Maria A., Favel, Anne
• Primary Examiner(s): Russel, Jeffrey E.
• Assistant Examiner(s): WALSH, STEPHEN G
• Application Number:
  US07/528,834
• Time in Patent Office: 635 Days
• Field of Search: 530/324, 514/12
• US Class Current: 514/20.4
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/81 Protease inhibitors

Abstract

A microprotein containing 28 amino acids of the formula ##STR1## wherein the X in position 4 is a residue of a member of the group consisting of valine, leucine, isoleucine, norleucine, alanine, cyclohexylalanine, o-methylthreonine, phenylglycine or α-amino butyric acid, Arginine, Y in position 5 is a residue of isoleucine or serine, Z in position 7 is a residue of methionine or norleucine, with the proviso that Z is a norleucine residue when X is an Arginine residue, having anti-elastase activity.

Claims

1. A polypeptide containing 28 amino acids of the formula ##STR4## wherein the X in position 4 is a residue of a member of the group consisting of valine, leucine, isoleucine, norleucine, alanine, cyclohexylalanine, o- methylthreonine, phenylglycine or α

-amino butyric acid, Arginine, Y in position 5 is a residue of isoleucine or serine, Z in position 7 is a residue of methionine or norleucine with the proviso that Z is a norleucine residue when X is an Arginine residue.

2. A polypeptide of claim 1 wherein Y in position 5 is an isoleucine residue.

3. A polypeptide of claim 1 wherein X in position 4 is selected from a residue of the group consisting of valine, alanine and norleucine.

4. A polypeptide of claim 1 wherein Z in position 7 is a norleucine residue.

5. A polypeptide of claim 1 wherein X in position 4 is a valine residue, Y in position 5 is an isoleucine residue and Z in position 7 is a norleucine residue.

6. A polypeptide of claim 1 wherein X in position 4 is an alanine residue, Y in position 5 is an isoleucine residue and Z in position 7 is a methionine residue.

7. A polypeptide of claim 1 wherein X in position 4 is a norleucine residue, Y in position 5 is a serine residue and Z in position 7 is a methionine residue.

8. A process for the preparation of a polypeptide of claim 1 comprising squentially introducing in a solid phase synthesis protected amine acids on a cross-linked oxymethyl polystyrene resin with a coupling agent, releasing the peptide chain formed from the resin, deprotecting the amino acids and cyclizing the 3 disulfide bridges to obtain the microprotein of claim 1.

9. The process of claim 8 wherein the cross-linked oxymethyl polystyrene resin is Boc-Gly-CM resin wherein Boc is a tert-butoxy carbonyl, Gly is glycine and CM is the cross-linked oxymethyl polystyrene resin.

10. The process of claim 8 wherein the coupling agent is BOP or (benzotriazol-1-yloxy) tris(dimethylamino) phosphonium hexafluorophosphate.

11. The process of claim 8 wherein the release from the resin and the deprotection of the amino acids are effected with hydrofluoric acid at about 0°

C.

12. An anti-elastase composition comprising an anti-elastase effective amount of at least one polypeptide of claim 1.

13. A composition of claim 12 wherein Y in the 5 position is an isoleucine residue.

14. A method of inducing anti-elastase activity in warm-blooded animals comprising administering to warm-blooded animals an anti-elastase effective amount of at least one polypeptide of claim 1.

15. A method of claim 14 wherein Y in position 5 is an isoleucine residue.

16. A method of claim 14 wherein X in position 4 is selected from a residue of the group consisting of valine, alanine and norleucine.

17. A method of claim 14 wherein Z in position 7 is a norleucine residue.

18. A method of claim 14 wherein X in position 4 is a valine residue, Y in position 5 is an isoleucine residue and Z in position 7 is a norleucine residue.

19. A method of claim 14 wherein X in position 4 is an alanine, residue, Y in position 5 is an isoleucine residue and Z in position 7 is a methionine residue.

20. A method of claim 14 wherein X in position 4 is a norleucine residue, Y in position 5 is a serine residue and Z in position 7 is a methionine residue.