Claims for Patent: 4,374,830

Summary for Patent: 4,374,830

Title:	Platelet aggregating material from equine arterial tissue
Abstract:	Novel hemostatic agent comprises equine arterial fibrillar collagen in a carrier. The agent is useful for the aggregation of platelets for clinical diagnostic tests and for the clotting of blood, such as for controlling bleeding in warm blooded species. The fibrillar collagen is obtained by extracting homogenized equine arterial tissue with aqueous solutions followed by extensive dialysis.
Inventor(s):	Schneider; Morris D. (Knoxville, TN)
Assignee:	Research Corp. (New York, NY)
Application Number:	06/292,201
Patent Claims:	1. A method of preparing a hemostatic composition comprising the steps of: (a) contacting cleaned, homogenized equine arterial tissue with a balanced salt solution to provide an aqueous extract containing extracted collagen material and unextracted arterial tissue, and (b) separating unextracted arterial tissue from said aqueous mixture to provide an extract containing an arterial collagen material, said extract being capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: 2. The method of claim 1 wherein the arterial tissue is burro aortic tissue. 3. The method of claim 1 wherein the arterial tissue is horse aortic tissue. 4. The method of claim 1 wherein the balanced salt solution is Tyrode's solution. 5. The method of claim 2 wherein the balanced salt solution is Tyrode's solution. 6. The method of claim 3 wherein the balanced salt solution is Tyrode's solution. 7. The method of claim 1 including the further step of (c) dialyzing the extract against water or a balanced salt solution to provide a substantially salt free retentate, said retentate being capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: 8. The method of claim 7 wherein the arterial tissue is burro aortic tissue. 9. The method of claim 7 wherein the arterial tissue is horse aortic tissue. 10. The method of claim 7 wherein Tyrode's solution is used in both Step (a) and (c). 11. The method of claim 7 including the further step of (d) ultracentrifuging said retentate to separate suspended material and resuspending said separated material in fresh balanced salt solution to provide a suspension, said suspension being capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: 12. The method of claim 11 wherein the arterial tissue is burro aortic tissue. 13. The method of claim 11 wherein the arterial tissue is horse aortic tissue. 14. The method of claim 11 wherein Tyrode's solution is used in Steps (a), (c) and (d). 15. The method of claim 11 including the further step of (e) dialyzing the suspension against water or a balanced salt solution to provide a substantially salt free retentate said retentate being capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin losing its platelet aggregating activity when exposed to collagenase, at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: 16. The method of claim 15 wherein the arterial tissue is burro aortic tissue. 17. The method of claim 15 wherein the arterial tissue is horse aortic tissue. 18. The method of claim 15 wherein Tyrode's solution is used in Steps (a), (b), (c) and (d). 19. A hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions which is an equine arterial fibrillar collagen capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: 20. A hemostatic agent of claim 19 in a pharmaceutically acceptable carrier. 21. A hemostatic agent of claim 19 in an externally administerable pharmaceutically acceptable carrier. 22. A hemostatic agent of claim 19 in a balanced salt solution. 23. A hemostatic agent of claim 19 in water. 24. A method of stimulating platelet aggregation in mammalian plasma or whole blood which comprises contacting said plasma or whole blood with an amount which is effective to stimulate platelet aggregation of a hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions which is an equine arterial fibrillar collagen capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent characterized by the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: 25. A method as in claim 24 wherein the hemostatic agent is in a pharmaceutically acceptable carrier. 26. A method on controlling external bleeding from a wound or incision comprising contacting said wound or incision with an amount which is effective to control bleeding of a hemostatic agent which is capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions as an equine arterial fibrillar collagen having the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: 27. A method of detecting abnormal platelet function comprising contacting platelets which are suspected of abnormal function in plasma or whole blood under test with a hemostatic agent which is capable of stimulating the aggregation of platelets in plasma or whole blood and containing a proteinaceous hemostatic agent isolatable from equine arterial tissue by extraction with aqueous solutions characterized as an equine arterial fibrillar collagen having the ability to enhance platelet aggregating activities in mammalian blood, being stable for extended periods of time at temperatures as low as -85.degree. C., stable at 56.degree. C. for up to 60 minutes, and when exposed to .alpha.-chymotrypsin, losing its platelet aggregating activity when exposed to collagenase at 37.degree. C. or when heated at 100.degree. C. for 15 minutes; containing a product having the following average number of amino acid residues per 1000 total amino acid residue: and comparing the aggregation response to the plasma or whole blood under study to the response of normal platelets.

Details for Patent 4,374,830

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2000-02-22