Claims for Patent: 5,686,262

Summary for Patent: 5,686,262

Title:	Recycle process for the production of low-cost soluble collagen
Abstract:	The invention includes a method that provides a low cost aqueous solution of solubilized collagen by the steps of: (a) providing an aqueous ground slurry of insoluble collagen and adjusting the pH of said slurry to obtain activity for a proteolytic enzyme added in Step b; (b) adding said proteolytic enzyme to said pH adjusted slurry; (c) reacting the slurry and enzyme of Step b and/or recycled insoluble collagen and enzyme from Step e at a temperature, T, and for a time, t, effective for forming a solution increased in solubilized collagen; (d) adding additional water and insoluble collagen to said solution of Step c and mixing; (e) separating at least some of the solution of Step d containing solubilized collagen from insoluble collagen, whereby at least a portion of said insoluble collagen and proteolytic enzyme is recycled to Step c, and the separated solution containing solubilized collagen is withdrawn as product; an alternative embodiment provides for the direct production of solubilized collagen without the recycle step.
Inventor(s):	Fink; David J. (Shaker Heights, OH), Brody; Richard S. (Worthington, OH)
Assignee:	Ranpak Corporation (Concord, OH)
Application Number:	08/488,368
Patent Claims:	1. A method for producing an aqueous solution of solubilized collagen comprising: a. providing an aqueous ground slurry of insoluble collagen and adjusting the pH of said slurry to obtain activity for a proteolytic enzyme added in Step b; b. adding said proteolytic enzyme to said pH adjusted slurry; c. reacting said slurry and enzyme of Step b and.backslash.or recycled insoluble collagen and enzyme of Step e at a temperature, T, and for a time, t, effective for forming a solution increased in solubilized collagen; d. adding insoluble collagen with or without additional water to said solution of Step c and mixing; e. separating at least some of the solution of Step d containing solubilized collagen from said insoluble collagen, whereby at least a portion of said insoluble collagen and proteolytic enzyme is recycled to Step c, and a separated solution containing solubilized collagen is withdrawn as product. 2. The method of claim 1, further comprising repeating Step c, Step d and Step e two or more times. 3. The method of claim 1, comprising a continuous process. 4. The method of claim 1, further comprising adding additional enzyme to the recycled insoluble collagen from Step e that substantially replaces enzyme removed with the withdrawal of product. 5. The method of claim 1, further comprising adding additional enzyme to the recycled insoluble collagen from Step e when the rate of reaction on recycling decreases below a predetermined level. 6. The method of claim 1, comprising stopping said reaction by (1) adjusting said pH to that where said proteolytic enzyme is substantially inactive; and/or (2) reducing the temperature to that where said proteolytic enzyme is substantially inactive. 7. The method of claim 1, further comprising in Step a, adjusting the liquid or solids content of said wet ground slurry so that said solids are adjusted to a concentration of about 0.1 to about 1.0 wt %. 8. The method of claim 1, whereby said temperature, T, comprises about 5.degree. C. to about 30.degree. C. 9. The method of claim 1, whereby said temperature, T, comprises about 15.degree. C. to about 28.degree. C. 10. The method of claim 7, whereby said solids concentration is between about 0.3 to 0.35 wt % and said reaction of Step c is at a temperature of about 10.degree. to about 30.degree. C., and for a time of 12 to 72 hours. 11. The method of claim 10, whereby said temperature is between 15.degree. C. and 28.degree. C. 12. The method of claim 1, further comprising: selecting said proteolytic enzyme from the group consisting of porcine mucosal pepsin, bromelain, chymopapain, chymotrypsin, collagenase, ficin, papain, peptidase, proteinase A, proteinase K, trypsin, microbial proteases and combinations of such enzymes. 13. The method of claim 1, further comprising: wherein said proteolytic is enzyme a microbial acid proteases. 14. The method of claim 12, further comprising: adjusting said pH to about 1.5-3.0, and said temperature to between about 18.degree. C. to about 28.degree. C., when porcine mucosal pepsin is selected. 15. The method of claim 1, further comprising: whereby at least 80 wt % of said insoluble collagen is converted to soluble collagen and the number average molecular weight is above 300,000 daltons. 16. The method of claim 1, further comprising: whereby at least 90 wt % of said insoluble collagen is converted to soluble collagen. 17. A method for producing an aqueous solution of solubilized collagen comprising: a. providing an aqueous ground slurry of insoluble collagen; b. adjusting the water or solid content of said wet ground slurry whereby said insoluble collagen is at a concentration that promotes substantially maximum solubilized collagen concentration and molecular weight in a final product; c. adjusting the pH of said slurry from Step b to obtain activity for a proteolytic enzyme added in Step d; d. adding and mixing said proteolytic enzyme with said pH adjusted slurry; e. reacting the slurry of Step d and.backslash.or said recycled insoluble collagen of Step g at a temperature, T, and for a time, t, effective for forming a solution comprising solubilized collagen derived from said insoluble collagen particles; f. adding insoluble collagen with or without additional water to said solution containing solubilized collagen in Step e and mixing; g. separating at least some of said solution of Step f containing solubilized collagen from said insoluble collagen and returning said insoluble collagen to Step e, whereby at least a portion of said proteolytic enzyme is recycled, and a separated solution containing solubilized collagen is withdrawn as product. 18. The method of claim 17, further comprising repeating Step e through Step f two or more times. 19. The method of claim 17, comprising stopping said reaction by (1) adjusting said pH to that where said proteolytic enzyme is substantially inactive; and/or (2) reducing the temperature to that where said proteolytic enzyme is substantially inactive. 20. The method of claim 17, further comprising adding additional enzyme to said insoluble collagen from Step e that substantially replaces enzyme removed with the withdrawal of product. 21. The method of claim 1, further comprising adding additional enzyme to said insoluble collagen from Step e when the rate of reaction on recycling decreases below a predetermined level. 22. The method of claim 17, further comprising in Step b, adjusting said liquid or solids content of said wet ground slurry so that said solids are adjusted to a concentration of about 0.1 to about 1.0 wt %. 23. The method of claim 17, whereby in Step e said temperature, T, comprises about 5.degree. C. to about 30.degree. C. 24. The method of claim 17, whereby in Step e said temperature, T, comprises about 15.degree. C. to about 28.degree. C. 25. The method of claim 17, whereby, in Step b, said solids concentration is between about 0.3 to 0.35 wt % and in Step e said reaction is at a temperature of about 10.degree. to about 30.degree. C., and for a time of 12 to 72 hours. 26. The method of claim 25, whereby said temperature is between 15.degree. C. and 28.degree. C. 27. The method of claim 17, further comprising: selecting said proteolytic enzyme from the group consisting of porcine mucosal pepsin, bromelain, chymopapain, chymotrypsin, collagenase, ficin, papain, peptidase, proteinase A, proteinase K, trypsin, microbial proteases, and combinations of such enzymes. 28. The method of claim 17, further comprising: wherein said proteolytic is enzyme a microbial acid protease. 29. The method of claim 27, further comprising in Step c adjusting said pH to about 1.5-3.0 and in Step e said temperature to between about 18.degree. C. to about 28.degree. C., when porcine mucosal pepsin is selected. 30. The method of claim 17, further comprising: whereby at least 80 wt % of said insoluble collagen is converted to soluble collagen and the number average molecular weight is above 300,000 daltons. 31. The method of claim 17, further comprising: whereby at least 90 wt % of said insoluble collagen is converted to soluble collagen. 32. A method for producing a solution of soluble collagen, said method comprising the steps of: a. reacting insoluble collagen with a proteolytic enzyme to produce a solution including soluble collagen; b. separating at least some of the soluble collagen from the insoluble collagen; c. withdrawing the soluble collagen as product; and d. returning the separated insoluble collagen to Step a, whereby at least a portion of the proteolytic enzyme is recycled. 33. A method as set forth in claim 32, whereby said insoluble collagen is provided as an aqueous ground slurry. 34. A method as set forth in claim 33, whereby the pH of said ground slurry is adjusted to provide activity for said proteolytic enzyme. 35. A method as set forth in claim 32, further comprising repeating Steps b, c and d two or more times. 36. A method as set forth in claim 32, further comprising adding additional insoluble collagen, with or without water or additional proteolytic enzyme, to the reacted solution of Step a, whereby said proteolytic enzyme reacts with said additional insoluble collagen. 37. A method as set forth in claim 32, further comprising adding additional water and proteolytic enzyme to Step a to the recycled insoluble collagen from Step d, so that enzyme removed with the withdrawal of said soluble collagen product is replaced. 38. A method as set forth in claim 32, further comprising adding additional proteolytic enzyme to the recycled insoluble collagen from Step d, when the rate of reaction on recycling decreases below a predetermined level. 39. A method as set forth in claim 32, wherein the insoluble collagen and proteolytic enzyme are reacted at a temperature, T, and for a time, t, effective for forming a solution increased in soluble collagen. 40. A method as set forth in claim 32, further comprising stopping said reaction by (a) adjusting the pH of said solution to that where said proteolytic enzyme is substantially inactive; and/or (b) reducing the temperature of said reaction to that where said proteolytic enzyme is substantially inactive. 41. A method as set forth in claim 33, further comprising adjusting the liquid or solids content of said aqueous ground slurry so that the solids concentration is between about 0.1 to about 1.0 wt %. 42. A method as set forth in claim 33, further comprising adjusting the liquids or solids content of said aqueous ground slurry so that the solids concentration is between about 0.3 to about 0.35 wt %. 43. A method as set forth in claim 41, wherein said temperature, T, is between about 5.degree. C. and about 30.degree. C. 44. A method as set forth in claim 41, wherein said temperature, T, is between about 15.degree. C. and about 28.degree. C. 45. A method as set forth in claim 44, wherein said temperature, T, is between about 10.degree. and about 30.degree. C., and said time, t, is about 12 to about 72 hours. 46. A method as set forth in claim 45, wherein said temperature, T, is between about 15.degree. C. and about 28.degree. C. 47. A method as set forth in claim 32, further comprising: selecting said proteolytic enzyme from the group consisting of porcine mucosal pepsin, bromelain, chymopapain, chymotrypsin, collagenase, ficin, papain, peptidase, proteinase A, proteinase K, trypsin, microbial proteases and combinations of such enzymes. 48. A method as set forth in claim 47, wherein said proteolytic enzyme is a microbial acid protease. 49. A method as set forth in claim 47, further comprising: adjusting pH to about 1.5 to about 3.0, and temperature to between about 18.degree. C. and about 28.degree. C., when porcine mucosal pepsin is selected. 50. A method as set forth in claim 32, whereby at least about 80 wt % of said insoluble collagen is converted to soluble collagen and the number average molecular weight of said soluble collagen is above about 300,000 daltons. 51. The method as set forth in claim 32, whereby at least about 90 wt % of said insoluble collagen is converted to soluble collagen. 52. The method as set forth in claim 33, further comprising adjusting the liquid or solids content of said aqueous ground slurry, whereby said insoluble collagen is at a concentration that promotes substantially maximum concentration and maximum molecular weight in said soluble collagen product.

Details for Patent 5,686,262

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Discure Medical, Llc	CHYMODIACTIN	chymopapain	For Injection	018663	11/10/1982		2014-11-11
Discure Medical, Llc	CHYMODIACTIN	chymopapain	For Injection	018663	08/21/1984		2014-11-11
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2014-11-11