Claims for Patent: 10,119,131

Summary for Patent: 10,119,131

Title:	Compositions and methods for producing clostridial collagenases
Abstract:	The present invention provides a method for producing a drug product comprising a combination of highly purified collagenase I and collagenase II from Clostridium histolyticum. The method utilizes an improved medium for the cultivation of Clostridium histolyticum which includes a non-meat-derived (i.e., non-mammalian) peptone or vegetable peptone. The method includes one or more of: (1) reducing glucose content in the meat-free or vegetable-derived media; and (2) increasing the salt concentration in the meat-free or vegetable-derived media. Also provided is a drug product which includes collagenase I and collagenase II at an optimized fixed mass ratio, and which has a purity of greater than at least 95%.
Inventor(s):	Wegman; Thomas L. (N. Merrick, NY), Yu; Bo (Fresh Meadows, NY)
Assignee:	BIOSPECIFICS TECHNOLOGIES CORP. (Lynbrook, NY)
Application Number:	13/422,939
Patent Litigation and PTAB cases:	See patent lawsuits and PTAB cases for patent 10,119,131
Patent Claims:	1. A method for fermenting a clostripain-producing Clostridium histolyticum to obtain a mixture of collagenase I and collagenase II, comprising fermenting said clostripain-producing Clostridium histolyticum in medium comprising peptone, wherein the peptone is Oxoid VG100 Vegetable Peptone made from pea, or Oxoid VG200 Vegetable Peptone phosphate broth, wherein said mixture of collagenase I and collagenase II is separately purified in the absence of added protease inhibitor and recombined to produce a drug product at least 95% pure as determined by reverse phase high performance liquid chromatography and contains no detectable clostripain activity using an enzymatic assay with N-a-Benzoyl-L-Arginine Ethyl Ester (BAEE) as substrate. 2. The method of claim 1, wherein the medium comprises greater than about 5 g/L total salt. 3. The method of claim 2, wherein the salt content in the medium is greater than about 7.5 g/L. 4. The method of claim 1, wherein the collagenase I and collagenase II are recombined at an optimized fixed mass ratio of 0.5 to 1.5. 5. The method of claim 4, wherein the optimized fixed mass ratio of collagenase I to collagenase II is 1. 6. The method of claim 1, wherein the collagenases are 97% pure. 7. The method of claim 1, wherein the collagenases are 98% pure. 8. The method of claim 1, wherein the purifying comprises: i. ammonium sulfate precipitation; ii. dialysis; iii. hydroxylapatite chromatography; iv. gel filtration; and v. anion exchange. 9. The method of claim 8, wherein steps (i), (ii), (iii), (iv) and (v) are performed in order. 10. The method of claim 1, wherein the clostripain-producing Clostridium histolyticum is ATCC 21000. 11. A method for fermenting a clostripain-producing Clostridium histolyticum to obtain a mixture of collagenase I and collagenase II, comprising fermenting said clostripain-producing Clostridium histolyticum in medium comprising peptone and lacking glucose, wherein the peptone is BBL Phytone Peptone, or BD Difco Select Phytone, wherein said mixture of collagenase I and collagenase II is separately purified in the absence of added protease inhibitor and recombined to produce a drug product at least 95% pure as determined by reverse phase high performance liquid chromatography and contains no detectable clostripain activity using an enzymatic assay with N-a-Benzoyl-L-Arginine Ethyl Ester (BAEE) as substrate. 12. The method of claim 11, wherein the collagenase I and collagenase II are recombined at an optimized fixed mass ratio of 0.5 to 1.5. 13. The method of claim 12, wherein the optimized fixed mass ratio of collagenase I to collagenase II is 1. 14. The method of claim 11, wherein the collagenases are 97% pure. 15. The method of claim 14, wherein the collagenases are 98% pure. 16. The method of claim 11, wherein the purifying comprises: i. ammonium sulfate precipitation; ii. dialysis; iii. hydroxylapatite chromatography; iv. gel filtration; and v. anion exchange. 17. The method of claim 16, wherein steps (i), (ii), (iii), (iv) and (v) are performed in order. 18. The method of claim 11, wherein the salt content in the medium is greater than about 7.5 g/L. 19. The method of claim 11, wherein the clostripain-producing Clostridium histolyticum is ATCC 21000.

Details for Patent 10,119,131

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2031-03-16