Claims for Patent: 5,756,454

Summary for Patent: 5,756,454

Title:	Collagen-induced platelet aggregation inhibitor
Abstract:	The invention provides a protein which inhibits collagen-induced platelet aggregation, derived from Triatoma pallidipennis, and derivatives thereof. The protein is useful for inhibition of collagen-induced human platelet aggregation or of cancer with metastatic tumor cells.
Inventor(s):	Noeske-Jungblut; Christiane (Berlin, MX), Haendler; Bernard (Berlin, MX), Kraetzschmar; Joern Reiner (Berlin, MX), Schleuning; Wolf-Dieter (Berlin, MX), Alagon; Alejandro (Cuernavaca, MX), Possani; Lourival (Cuernavaca, MX), Cuevas-Aguirre; Delia (Cuernavaca, MX)
Assignee:	Schering Aktiengesellschaft (Berlin, DE)
Application Number:	08/206,185
Patent Claims:	1. An isolated protein having the N-terminal amino acid sequence: ##STR9## wherein the protein is encoded by a DNA molecule having a sequence which hybridizes with a DNA molecule of SEQ ID NO: 14, 15 or 16 under conditions wherein hybridization is conducted at 50.degree. C. in the presence of 5 .times. SSC, 5 .times. Denhardt's solution, and 100 .mu.g/ml salmon sperm DNA, and washing is conducted at 50.degree. C. in 2 .times. SSC and 0.2% SDS, and wherein said protein inhibits collagen-induced aggregation of mammalian platelets. 2. An isolated protein which inhibits collagen-induced aggregation of mammalian platelets having the following amino acid sequence: a) the sequence in i) SEQ ID NO:11, ii) SEQ ID NO:12 or iii) SEQ ID NO;13, or b) an amino acid sequence encoded by a DNA molecule having a sequence which hybridizes with a DNA molecule of SEQ ID NO: 14, 15 or 16 under conditions wherein hybridization is conducted at 50.degree. C. in the presence of 5 .times. SSC, 5 .times. Denhardt's solution, and 100 .mu.g/ml salmon sperm DNA, and washing is conducted at 50.degree. C. in 2 .times. SSC and 0.2% SDS, or c) a protein comprising a sequence of SEQ ID NO:11, 12 or 13 or a sequence as in b), and further comprising a posttranslational modification which does not substantially reduce the activity of the mature protein. 3. A protein of claim 1 which inhibits tumor cell-collagen interaction. 4. A protein of claim 2, which is recombinantly produced. 5. A protein of claim 4, which is unglycosylated. 6. A protein of claim 1, wherein said protein (a) is not fibrinogen-receptor antagonist; (b) is not a thromboxane-antagonist; (c) inhibits release-reaction of collagen-treated platelets; (d) does not inhibit platelet aggregation induced by thrombin or ADP; (e) does not react with collagen under in vivo conditions; (f) has its platelet aggregation inhibition reversed by a large amount of collagen under in vitro conditions; (g) has an inhibitory activity which is not influenced by protease-inhibitors; (h) has an inhibitory activity which increases in the presence of Mg.sup.2+ ions; and (i) is not cleavable by collagenase. 7. A protein of claim 6 which is a collagen receptor antagonist. 8. A protein of claim 1 which does not react with or bind to collagen in vivo. 9. A pharmaceutical composition comprising a protein of claim 1 and a pharmaceutically acceptable carrier. 10. A method of inhibiting collagen-induced human platelet aggregation comprising administering an effective amount of protein of claim 1. 11. A method of claim 10 for treating an atherosclerotic or thrombotic disease lesions or for preventing reocclusion after treatment of myocardial infarction. 12. A method of inhibiting tumor cell-collagen interaction comprising administering an effective amount of protein of claim 1. 13. A method of claim 12 for treating or preventing tumor cell metastasis. 14. A protein of claim 1 having an IC.sub.50 for collagen-induced aggregation of at least 2.5 .mu.g. 15. A collagen-induced platelet aggregation inhibitor protein having the amino acid sequence of SEQ ID NO:11, 12 or 13. 16. A protein of claim 15, produced by expression of a recombinant DNA sequence. 17. A protein of claim 16, wherein the DNA has the sequence of SEQ ID NO:14, 15 or 16. 18. A protein of claim 16, further comprising a precursor sequence, and said preprotein having the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:10. 19. A preprotein of claim 18, produced by expression of a recombinant DNA sequence. 20. A preprotein of claim 19, wherein the DNA has the sequence SEQ ID NO: 17, 18 or 19. 21. A protein of claim 15, produced by cleavage of a preprotein. 22. A method of purifying a protein of claim 1, comprising separating the protein on a gel filtration column according to molecular size filtration, using no salt gradient; and chromatographing the protein on a high performance electrophoresis chromatography system. 23. A protein of claim 1, which is recombinantly produced. 24. A method of purifying a protein of claim 23, comprising separating the protein on a gel filtration column according to molecular size filtration, using no salt gradient; and chromatographing the protein on a high performance electrophoresis chromatography system. 25. A protein of claim 1, wherein said protein (a) is not fibrinogen-receptor antagonist; (b) is not a thromboxane-antagonist; (c) inhibits release-reaction of collagen-treated platelets; (d) does not inhibit platelet aggregation induced by thrombin or ADP; (e) does not react with collagen under in vivo conditions; (f) has its platelet aggregation inhibition reversed by a large amount of collagen under in vitro conditions; (g) has an inhibitory activity which is not influenced by protease-inhibitors; (h) has an inhibitory activity which increases in the presence of Mg.sup.2+ ions; or (i) is not cleavable by collagenase. 26. An isolated protein of claim 1, wherein the DNA molecule encodes a naturally occurring protein. 27. An isolated protein of claim 2, wherein the DNA molecule encodes a naturally occurring protein. 28. A pharmaceutical composition of claim 9, wherein the DNA molecule encodes a naturally occurring protein. 29. A method of claim 10, wherein the DNA molecule encodes a naturally occurring protein. 30. A method of claim 12, wherein the DNA molecule encodes a naturally occurring protein.

Details for Patent 5,756,454

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2015-05-26