You’re using a public version of DrugPatentWatch with 5 free searches available | Register to unlock more free searches. CREATE FREE ACCOUNT

Last Updated: March 29, 2024

Claims for Patent: 9,738,883


✉ Email this page to a colleague

« Back to Dashboard


Summary for Patent: 9,738,883
Title:Process for the production and purification of the collagenase enzyme from vibrio alginolyticus
Abstract: The present invention claims a novel process for the production and purification of microbial collagenase (Microbial Collagenase EC 3.4.24.3) produced by the non-pathogenic aerobic bacterium Vibrio alginolyticus chemovar. iophagus (NCIMB Number: 1 1038, synonym LMG 3418, hereinafter called Vibrio alginolyticus), which said process provides high production levels of collagenase with a stable, reproducible, cheap fermentation process. The collagenase produced from Vibrio alginolyticus according to the process described herein also presents a specific activity superior to that of other microbial collagenases, is stable in aqueous solution, and can be frozen without significant damage. A further subject of the present invention is pharmaceutical compositions containing collagenase obtained according to the production and purification process described, for the purpose of therapeutic treatment of disorders characterized by collagen accumulation or for the treatment of blemishes/imperfections that benefit from reducing local collagen accumulations.
Inventor(s): Vaccaro; Susanna (Abano Terme, IT), Caputo; Michele (Abano Terme, IT), Cuppari; Christian (Abano Terme, IT), Gennari; Giovanni (Abano Terme, IT)
Assignee: FIDIA FARMACEUTICALS S.P.A. (Abano Terme, IT)
Application Number:14/394,481
Patent Claims:1. A process for the production and purification of collagenase from Vibrio alginolyticus chemovar. lophagus, comprising the following stages: Stage A: Inoculation of Vibrio Alginolyticus chemovar. lophagus into an Erlenmeyer flask and fermentation with culture broth of non-bovine animal origin; Stage B: Clarification of the fermented broth thus obtained by tangential flow ultrafiltration with 100-500 kD Molecular Weight Cut-Off (MWCO) cassettes; Stage C: Dialysis and concentration of the clarified medium obtained in stage B, by tangential flow ultrafiltration with 5-30 kD MWCO cassettes; Stage D: Purification of the solution containing collagenase obtained in Stage C, by anion-exchange resin carrying weak basic groups, at a pH of between 6.9 and 7.4; Stage E: Dialysis and concentration of the fractions with collagenolytic activity collected in Stage D, by tangential flow ultrafiltration with 10-50 kD MWCO cassettes; Stage F: Purification of the solution thus obtained, by anion-exchange resin carrying strong basic groups, at a pH of between 6.9 and 7.4; Stage G: Diafiltration and concentration of the fractions with collagenolytic activity .gtoreq.95% originating from stage F, by tangential flow ultrafiltration with 10-50 kD MWCO cassettes; Stage H: Filtration of the solution containing collagenase thus obtained, through an 0.2 .mu.m absolute filter, and storage at a temperature of between -20.degree. and -80.degree. C.; wherein the thus obtained collagenase from Vibrio alginolyticus chemovar. lophagus being characterized by: molecular weight 82 Kda; specific activity between 1000 and 1800 nkat/mg; purity between 98.0 and 100%; no microbial or protein contaminants; stability at a pH of between 5.5 and 11; stability in aqueous solution at a temperature ranging between 4.degree. and 40.degree. C.; stability in aqueous solution at 4.degree. C. for 30 days; stability in aqueous solution at a temperature ranging between -20.degree. C. and -80.degree. C. for 24-48 months; lyophilisability to obtain a stable lyophilic powder; and caseinase activity is less than 1.0 U/ml.

2. The process according to claim 1, wherein the culture broth is of porcine animal origin or is a mixture of porcine and plant origin.

3. The process according to claim 1 or 2, wherein the anion-exchange resin carrying the weak basic groups carries diethylaminoalkyl groups and the anion-exchange resin carrying strong basic groups carries quaternary ammonium groups.

Make Better Decisions: Try a trial or see plans & pricing

Drugs may be covered by multiple patents or regulatory protections. All trademarks and applicant names are the property of their respective owners or licensors. Although great care is taken in the proper and correct provision of this service, thinkBiotech LLC does not accept any responsibility for possible consequences of errors or omissions in the provided data. The data presented herein is for information purposes only. There is no warranty that the data contained herein is error free. thinkBiotech performs no independent verification of facts as provided by public sources nor are attempts made to provide legal or investing advice. Any reliance on data provided herein is done solely at the discretion of the user. Users of this service are advised to seek professional advice and independent confirmation before considering acting on any of the provided information. thinkBiotech LLC reserves the right to amend, extend or withdraw any part or all of the offered service without notice.