| PATENT NUMBER | This data is not available for free |
| PATENT GRANT DATE | December 3, 1996 |
| PATENT TITLE |
Amylin antagonist peptides and uses therefor |
| PATENT ABSTRACT | Compounds which inhibit amylin activity are provided. These compounds may be used in the treatment of conditions where it is of benefit to reduce amylin activity, including the treatment of Type 2 diabetes mellitus, impaired glucose tolerance, obesity and insulin resistance |
| PATENT INVENTORS | This data is not available for free |
| PATENT ASSIGNEE | This data is not available for free |
| PATENT FILE DATE | November 19, 1991 |
| PATENT REFERENCES CITED |
Breimer et al. Biochem. J., 255, 377-390, 1988. Lehninger, A., Principles of Biochemistry, pp. 100-103. Leighton, B., et al., Nature, vol. 335, No. 6191, pp. 632-635 (13 Oct. 1988). Cooper, G. J. S., et al. Proc. Natl. Acad. Sci USA, vol. 85, pp. 7763-7766 (Oct. 1988). Roberts, A. N., et al., Proc. Natl. Acad. Sci USA, vol. 86, pp. 9662-9666 (Dec. 1989). Munson, P. J., et al., Analytical Biochemistry, 107, pp. 220-239 (1980). Cooper, G. J. S., et al., Biochimica et Biophysica Acta., 1014, pp. 247-258 (1989). |
| PATENT PARENT CASE TEXT | This data is not available for free |
| PATENT CLAIMS |
We claim: 1. An amylin antagonist peptide which is an N-terminal deletion peptide of Peptide A, modified Peptide A, Peptide B, modified Peptide B, Peptide C, or modified Peptide C, wherein at least the first two to seven N-terminal amino acid residues of Peptide A or modified Peptide A have been deleted, at least the first eight N-terminal amino acid residues of Peptide B or modified Peptide B have been deleted, and at least the first two to seven N-terminal amino acid residues of Peptide C or modified Peptide C have been deleted, each such amylin antagonist peptide optionally having an acetylated N-terminal amino acid, a carboxy-amidated C-terminal amino acid, or both. 2. An amylin antagonist peptide according to claim 1 having an IC.sub.50 in an amylin receptor assay of less than about 5 nM and an IC.sub.50 in a soleus muscle antagonist assay of less than about 1 .mu.M. 3. An amylin antagonist peptide according to claim 1 which is a deletion peptide based on Peptide C or modified Peptide C. 4. An amylin antagonist peptide according to claim 3 wherein said deletion peptide based on Peptide C or modified Peptide C is .sup.8-32 Peptide C or modified .sup.8-32 Peptide C or a peptide having successive N-terminal deletions thereof up to and including .sup.24-32 Peptide C or modified .sup.24-32 Peptide C. 5. An amylin antagonist peptide according to claim 4 wherein modified Peptide C is a peptide having at least one amino acid substitution selected from the group .sup.26 Ala, .sup.26 Asp, .sup.26 Gln, .sup.26 Glu, .sup.27 Val, .sup.29 Ala, .sup.29 Gly, .sup.30 Asn, .sup.30 Lys, .sup.30 Arg, .sup.30 Ala, .sup.30 Phe, .sup.32 Tyr, .sup.32 Hyp, .sup.32 Thr, .sup.32 Phe or .sup.32 Hydroxyproline. 6. An amylin antagonist peptide according to claim 5 which is modified .sup.8-32 Peptide C, modified .sup.9-32 Peptide C, modified .sup.22-32 Peptide C or modified .sup.24-32 Peptide C. 7. An amylin antagonist peptide according to claim 5 which is modified .sup.8-32 Peptide C or modified .sup.9-32 Peptide C, said modified Peptides not having a C-terminal NH.sub.2 group. 8. An amylin antagonist peptide according to claim 5 which is modified .sup.22-32 Peptide C or modified .sup.24-32 Peptide C, said modified Peptides not having a C-terminal NH.sub.2 group. 9. An amylin antagonist peptide according to claim 3 which is .sup.9-32 Peptide C or modified .sup.9-32 Peptide C. 10. An amylin antagonist peptide according to claim 9 wherein modified Peptide C is a peptide having at least one amino acid substitution selected from the group consisting of .sup.11 Arg, .sup.15 Leu, .sup.18 Arg, .sup.30 Asn, and .sup.32 Tyr. 11. An amylin antagonist peptide according to claim 3 which is from about 9 to about 11 amino acids long. 12. An amylin antagonist peptide according to claim 11 which is .sup.22-32 Peptide C, modified .sup.23-32 Peptide C, .sup.24-32 Peptide C or modified .sup.26-32 Peptide C. 13. An amylin antagonist peptide according to claim 12 having at least one amino acid substitution selected from the group consisting of .sup.22 D-Nal, .sup.22 L-Nal, .sup.26 D-Asp, .sup.26 L-Asp, .sup.27 Val, .sup.30 Asn, .sup.30 Phe, .sup.30 Lys, .sup.30 Arg, .sup.32 Tyr and .sup.32 Hydroxyproline. 14. An amylin antagonist peptide which is an N-terminal deletion peptide of Peptide C having at least amino acids 1 to 7 deleted and optionally including substitutions in amino acid sequence selected from: at residue 11, replacing Lys with Arg; at residue 15, replacing Glu with Phe; at residue 18, replacing Lys with Arg; at residue 26, replacing Asn with Ala, Asp, Gln or Glu; at residue 27, replacing Thr with Val; at residue 29, replacing Ser with Ala or Gly; at residue 30, replacing Glu with Asn, Lys, Arg or Ala; and at residue 32, replacing Pro with Tyr, Phe or Hydroxyproline; and an optionally having an acetylated N-terminal amino acid, a carboxy-amidated C-terminal amine acid, or both. 15. An amylin antagonist peptide according to claim 14 having the amino acid sequence Ac-.sup.11 Arg.sup.15 Leu.sup.18 Arg.sup.30 Asn.sup.32 Tyr.sup.9-32 Peptide C. 16. An amylin antagonist peptide according to claim 14 having the amino acid sequence Ac-.sup.11 Arg.sup.18 Arg.sup.30 Asn.sup.32 Tyr.sup.9-32 Peptide C. 17. An amylin antagonist peptide according to claim 14. 18. An amylin antagonist peptide according to claim 14 having the amino acid sequence Ac-.sup.30 Asn.sup.32 Tyr.sup.8-32 Peptide C. 19. An amylin antagonist peptide of claim 1 having the amino acid sequence X-Xaa.sub.1 -Leu-Gly-Xaa.sub.2 -Leu-Ser-Gln-Xaa.sub.3 -Leu-His-Xaa.sub.4 -Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Xaa.sub.5 -Gly-Xaa.sub.6 -Xaa.sub.7 -Thr-Xaa.sub.8 -Y wherein X is lower acyl or hydrogen; Xaa.sub.1 is Val, Ala or absent; Xaa.sub.2 is Lys, Arg or Orn; Xaa.sub.3 is Glu, Asp, Leu or Phe; Xaa.sub.4 is Lys, Arg or Orn; Xaa.sub.5 is Thr or Val; Xaa.sub.6 is Ser or Ala; Xaa.sub.7 is Gly, Asn, Lys, Arg or Orn; Xaa.sub.8 is Pro, Tyr or Phe; and Y is --OH or --NH.sub.2. 20. An amylin antagonist peptide of claim 19 wherein Xaa.sub.5 is Thr and Xaa.sub.6 is Ser. 21. An amylin antagonist peptide of claim 20 wherein Xaa.sub.7 is Asn and Xaa.sub.8 is Tyr. 22. An amylin antagonist peptide of claim 21 wherein Xaa.sub.3 is Glu. 23. An amylin antagonist peptide of claim 22 wherein Xaa.sub.1 is Val. 24. An amylin antagonist peptide of claim 22 wherein Xaa.sub.1 is absent. -------------------------------------------------------------------------------- |
| PATENT DESCRIPTION |
DESCRIPTION OF THE INVENTION This invention is directed to compounds which inhibit amylin activity. These compounds may be used in the treatment of Type 2 diabetes mellitus and other disorders, including obesity, insulin resistance, impaired glucose tolerance, disorders involving excess amylin action, and other disorders where amylin activity is benefically reduced. DESCRIPTION OF RELATED ART AND INTRODUCTION TO THE INVENTION The present invention is directed to compounds which inhibit amylin activity and their use as therapeutic agents for type 2 diabetes and other disorders. Amylin is a newly discovered peptide which has marked effects on carbohydrate metabolism in vitro and in vivo including the ability to inhibit the uptake of glucose into glycogen, and the promotion of glycogenolysis in isolated skeletal muscle. Cooper, G. J. S., et al., Proc. Natl. Acad. Sci. USA 85: 7763-7766 (1988). A defect in amylin homeostasis is believed to contribute to insulin resistance and the development of type 2 diabetes, Cooper, G. J. S., et al., Biochim. Biophys. Acta. 1014: 247-258 (1989), as well as other metabolic disorders. Amylin is 37 amino acids in length (see FIG. 1), and requires both an intact intramolecular disulfide bond and a C-terminal amide to exert its full biological activity on glycogen synthesis in skeletal muscle. E.g., Roberts, A. N., et al., Proc. Natl. Acad. Sci. USA 86: 9662-9666 (1989). SUMMARY OF THE INVENTION The present invention is directed to novel compounds which regulate the effects of amylin and amylin-like compounds (the latter compounds are also referred to as "amylin agonists"). These compounds inhibit the effects of amylin and its agonists, and are referred to as "amylin inhibitors" or amylin "antagonists." Several assays have been developed to measure amylin activity and to evaluate new compounds. A receptor binding assay as described herein may be used to screen for or to identify compounds which bind to amylin receptors and which may, therefore, be candidate amylin agonists and antagonists. The rat soleus muscle assay may be used as a secondary procedure to screen for or to evaluate and differentiate between amylin agonists and amylin antagonists. In the soleus muscle assay, amylin and amylin agonists inhibit insulin-stimulated glycogenesis. Amylin antagonists counteract this amylin inhibition which results in the recovery of insulin-stimulated glycogenesis. The amylin inhibitors of this invention may be subdivided into three major categories: (i) truncated peptides, (ii) structurally constrained peptides, and (iii) peptides having amino acid substitutions including the substitution of unusual or unnatural amino acids for one or more of the amino acids in the naturally occurring peptide sequence, as well as peptides having a combination of the modifications (i), (ii) and/or (iii). Three different peptides were used in the preparation of compounds within these categories and are hereforth referred to as Peptides A, B and C. The sequences of these peptides are depicted in FIG. 2. |
| PATENT PHOTOCOPY | available on request |
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