Calcium channel blocking polypeptides from filistata hibernalis

Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai

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530324, A61K 3800, C07K 500, C07K 700

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058045546

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BRIEF SUMMARY
This is a national stage application, filed pursuant to 35 U.S.C. .sctn.371, of PCT international application Ser. No. PCT/US93/03921, filed Apr. 30, 1993, published as WO93/23428 Nov. 25, 1993.


BACKGROUND OF THE INVENTION

This invention relates to polypeptides found in the venom of the Filistata hibernalis spider and to polypeptides. having substantially the same amino acid sequence and substantially the same activity as said polypeptides. The polypeptides and the pharmaceutically acceptable salts thereof block calcium channels in cells including neuronal and muscle cells of various organisms including invertebrates and vertebrates. This invention also relates to the use of said polypeptides and their salts in blocking calcium channels in cells such as cells in the nervous and muscular system of an organism, per se, and in the treatment of calcium channel mediated diseases and conditions in a mammal. Further, this invention relates to compositions comprising said polypeptides and salts thereof.
Compounds which are calcium antagonists have a variety of utilities. Calcium antagonists can find clinical application in the treatment of such conditions as angina, hypertension, cardiomyopathies, supraventricular arrhythmias, aesophogeal achalasia, premature labor and Raynaud's disease among others. See W. G. Nayler, Calcium Antagonists, Academic Press, Harcourt Brace Jovanovich Publishers, New York, N.Y. 1988, the teachings of which are incorporated herein by reference. Further, such compounds are useful in the study of the physiology of cells such as neuronal and muscle cells.


SUMMARY OF THE INVENTION

This invention concerns polypeptides found to be present in the venom of the Filistata hibernalis spider. The polypeptides of this invention and the fractions in which they are present according to this invention are as follows.
I. Filistata peptide 10 has the following N-terminal amino acid sequence, SEQ ID NO:1. N-Ala-Glu-Cys-Val-Asn-lle-Tyr-Gln-Pro-Cys-Ser-Thr-lle-Gly-Leu-Arg-Cys-Cys- Tyr-Gly- Ala-Arg-Cys-Tyr-Cys-Lys-Glu-Lys-Leu-Asn-Cys-Arg-Tyr-Asn-Arg-Ser-Thr-Arg-Ly s-Arg- Asp-Cys-Gly-Trp-Ser-Ser-Tyr-Asp-Cys-Lys-Cys-Asp-Tyr-Thr-Trp-Met-His-Arg-ll e-Asp- Asp-Trp-Arg-Glu-Gly-Tyr-Ser-Cys-Tyr-Cys-Lys-Glu-CO.sub.2 H
II. Filistata peptide 12 has the following N-terminal amino acid sequence, SEQ ID NO:2. N-Ala-Glu-Cys-Leu-Met-Val-Gly-Asp-Thr-Ser-Cys-Val-Pro-Arg-Leu-Gly-Arg-Arg- Cys- Cys-Tyr-Gly-Ala-Trp-Cys-Tyr-Cys-Asp-Gln-Gln-Leu-Ser-Cys-Arg-Arg-Val-Gly-Ar g-Lys- Gln-Gln-Cys-Gly-Trp-Arg-Glu-Val-Asn-Cys-Lys-Cys-Asp-Trp-Ser-Trp-Ser-Gln-Ar g-lle- Asp-Asp-Trp-Arg-Ala-Asp-Tyr-Ser-Cys-Lys-Cys-Pro-Glu-Asp-Gln-CO.sub.2 H
III. Filistata peptide 13-1 has the following N-terminal amino acid sequence, SEQ ID NO:3. N-Ala-Glu-Cys-Leu-Met-Val-Gly-Asp-Thr-Ser-Cys-Val-Pro-Arg-Leu-Gly-Arg-Arg- Cys Cys-Tyr-Gly-Ala-Trp-Cys-Tyr-Cys-Asp-Gln-Gln-Leu-Ser-Cys-Arg-Arg-Val-Gly-Ar g-Lys- Arg-Glu-Cys-Gly-Trp-Val-Glu-Val-Asn-Cys-Lys-Cys-Gly-Trp-Ser-Trp-Ser-Gln-Ar g-lle- Asp-Asp-Trp-Arg-Ala-Asp-Tyr-Ser-Cys-Lys-Cys-Pro-Glu-Asp-Gin-CO.sub.2 H
IV. Filistata peptide 13-2 has the following N-terminal amino acid sequence, SEQ ID NO:4. N-Ala-Glu-Cys-Leu-Met-Val-Gly-Asp-Thr-Ser-Cys-Val-Pro-Arg-Leu-Gly-Arg-Arg- Cys- Cys-Tyr-Gly-Ala-Trp-Cys-Tyr-Cys-Asp-Gln-Gln-Leu-Ser-Cys-Arg-Arg-Val-Gly-Ar g-Lys- Arg-Glu-Cys-Gly-Trp-Val-Glu-Val-Asn-Cys-Lys-Cys-Gly-Trp-Ser-Trp-Ser-Gln-Ar g-lle- Asp-Asp-Trp-Arg-Ala-Asp-Tyr-Asn-Cys-Lys-Cys-Pro-Glu-Asp-Gln-CO.sub.2 H
V. Filistata peptide 13-3 has the following N-terminal amino acid sequence, SEQ ID NO:5. N-Ala-Glu-Cys-Leu-Met-lie-Gly-Asp-Thr-Ser-Cys-Val-Pro-Arg-Leu-Gly-Arg-Arg- Cys- Cys-Tyr-Gly-Ala-Trp-Cys-Tyr-Cys-Asp-Gln-Gln-Leu-Ser-Cys-Arg-Arg-Val-Gly-Ar g-Lys- Arg-Glu-Cys-Gly-Trp-Val-Glu-Val-Asn-Cys-Lys-Cys-Gly-Trp-Ser-Trp-Ser-Gln-Ar g-lle- Asp-Asp-Trp-Arg-Ala-Asp-Tyr-Ser-Cys-Lys-Cys-Pro-Glu-Asp-Gln-CO.sub.2 H
VI. Filistata peptide 13-4 has the following N-terminal amino acid sequence, SEQ ID NO:6. N-Ala-Glu-Cys-Val-Asn-lle-Tyr-Gln-Pro-Cys-Ser-Asn-lle-Gly-Leu-Arg-Cys-Cys- Tyr- Gly-Ala-Arg-Cys-Tyr-Cys-Lys-Glu-Lys-Leu-S

REFERENCES:
Adams et al., J of Biol. Chem., vol. 265, No. 2, pp. 861-867, Jan. 15, 1990 .

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