Compositions and methods for treating collagen-mediated diseases
AssigneeAuxilium US Holdings, LLC
InventorsGregory L. Sabatino, Benjamin J. Del Tito, Jr., Phillip J. Bassett, Hazel A. Tharia, Antony G. Hitchcock
Life of this patent
Estimating the term…
Abstract
A drug product comprising a combination of highly purified collagenase I and collagenase II from Colostridium histolyticum is disclosed. The drug product includes collagenase I and collagenase II in a ratio of about 1 to 1, with a purity of greater than at least 95%. The invention further disclosed improved fermentation and purification processes for preparing the said drug product.
Claims29
A drug product consisting of isolated and purified collagenase I and collagenase II having the sequence of Clostridium histolyticum collagenase I and collagenase II, respectively, wherein the collagenase I and collagenase II have a mass ratio of about 1 to 1 and the drug product is at least 97% by area pure as determined by reverse phase high performance liquid chromatography.
A drug product consisting of isolated and purified collagenase I and collagenase II having the sequence of Clostridium histolyticum collagenase I and collagenase II, respectively, wherein the collagenase I and collagenase II have a mass ratio of about 1 to 1 and the drug product is at least 97% by area pure as determined by reverse phase high performance liquid chromatography, wherein the preparation of the drug product comprises the steps of:
- a) fermenting Clostridium histolyticum;
- b) harvesting a crude fermentation comprising collagenase I and collagenase II;
- c) purifying collagenase I and collagenase II from the crude harvest via filtration and column chromatography comprising the steps of:
- i) filtering the crude harvest through an anion exchange filter;
- ii) adding ammonium sulphate;
- iii) subjecting the harvest through a HIC column;
- iv) adding leupeptin to the filtrate;
- v) removing the ammonium sulfate;
- vi) filtering the mixture of step (v);
- and
- vii) separating collagenase I and collagenase II using ion-exchange;
- d) combining the collagenase I and collagenase II purified from step (c) at a ratio of about 1 to 1.
A process for producing a drug product consisting of isolated and purified collagenase I and collagenase II having the sequence of Clostridium histolyticum collagenase I and collagenase II, respectively, wherein the collagenase I and collagenase II have a mass ratio of about 1 to 1 and the drug product is at least 97% by area pure as determined by reverse phase high performance liquid chromatography, comprising the steps of:
- a) fermenting Clostridium histolyticum;
- b) harvesting a crude fermentation comprising collagenase I and collagenase II;
- c) purifying collagenase I and collagenase II from the crude harvest via filtration and column chromatography comprising the steps of:
- i) filtering the crude harvest through an anion exchange filter;
- ii) adding ammonium sulphate;
- iii) subjecting the harvest through a HIC column;
- iv) adding leupeptin to the filtrate;
- v) removing the ammonium sulfate;
- vi) filtering the mixture of step (v);
- and
- vii) separating collagenase I and collagenase II using ion-exchange;
- d) combining the collagenase I and collagenase II purified from step (c) at a ratio of about 1 to 1.
A pharmaceutical formulation comprising a pharmaceutically acceptable excipient and a drug product consisting of isolated and purified collagenase I and collagenase II having the sequence of Clostridium histolyticum collagenase I and collagenase II, respectively, wherein the collagenase I and collagenase II have a mass ratio of about 1 to 1 and the drug product is at least 97% by area pure as determined by reverse phase high performance liquid chromatography.
A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a drug product consisting of isolated and purified collagenase I and collagenase II having the sequence of Clostridium histolyticum collagenase I and collagenase II, respectively, wherein the collagenase I and collagenase II have a mass ratio of about 1 to 1 and the drug product is at least 97% by area pure as determined by reverse phase high performance liquid chromatography, wherein the preparation of the drug product comprises the steps of:
- a) fermenting Clostridium histolyticum;
- b) harvesting a crude fermentation comprising collagenase I and collagenase II;
- c) purifying collagenase I and collagenase II from the crude harvest via filtration and column chromatography comprising the steps of:
- i) filtering the crude harvest through an anion exchange filter;
- ii) adding ammonium sulphate;
- iii) subjecting the harvest through a HIC column;
- iv) adding leupeptin to the filtrate;
- v) removing the ammonium sulfate;
- vi) filtering the mixture of step (v);
- and
- vii) separating collagenase I and collagenase II using ion-exchange;
- d) combining the collagenase I and collagenase II purified from step (c) at a ratio of about 1 to 1.
A drug product consisting of collagenase I and collagenase II, wherein the collagenase I and collagenase II are isolated and purified from Clostridium histolyticum and wherein the collagenase I and collagenase II have a mass ratio of about 1 to 1 and the drug product is at least 97% by area pure as determined by reverse phase high performance liquid chromatography.
Dependencies are inferred from the claim text.