Animal product free media and processes for obtaining abotulinumtoxin

• US 7,148,041 B2
• Filed: 09/25/2003
• Issued: 12/12/2006
• Est. Priority Date: 09/25/2003
• Status: Active Grant

• Current Assignee: Allergan, Inc. (Canada) (Abbvie Incorporated)
• Original Assignee: Allergan, Inc. (Canada) (Abbvie Incorporated)
• Inventors: Donovan, Stephen
• Primary Examiner(s): KAM, CHIH MIN
• Application Number:
  US10/672,876
• Publication Number:
  US 20050069562A1
• Time in Patent Office: 1,174 Days
• Field of Search: 435/71.1, 435/252.7, 435/253.6, 424/842, 424/236.1, 424/239.1
• US Class Current: 435/71.1
• CPC Class Codes: A61P 1/10 Laxatives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/06 Antimigraine agents
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 9/52 derived from bacteria or Archaea
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione
• CPC Class Codes: C12Y 304/24069 Bontoxilysin (3.4.24.69), i.e. botulinum neurotoxin
• CPC Class Codes: Y10S 435/842 Clostridium

Abstract

Media and processes for the fermentation of Clostridium botulinum and obtaining a botulinum toxin for use in formulating botulinum toxin pharmaceutical compositions. The growth media can contain significantly reduced levels of meat or dairy by-products using non-animal based products to replace the animal-derived products. Preferably, the media used are substantially free of animal derived products.

Claims

1. A method for obtaining a biologically active botulinum toxin, comprising the steps of:

(a) providing a fermentation medium which is free of an animal product;

(b) culturing a Clostridium botulinum bacterium in the fermentation medium under conditions which permit production of a botulinum toxin, and;

(c) recovering a biologically active botulinum toxin from the fermentation medium,

wherein the fermentation medium comprises a protein product obtained from yeast or from a vegetable, and wherein the vegetable is selected from the group consisting of soy, malt and corn.

2. The method of claim 1, wherein in the step of culturing, the culturing is performed until at least 48 hours after initial drop in cell density due to cell lysis.

3. The method of claim 1, wherein the botlinumtoxin is selected from the group consisting of botulinum toxins types A, B, C, D, E, F and G.

4. The method of claim 1, wherein the botulinum toxin is botulinum toxin types A.

5. The method of claim 1, wherein the botulinum toxin is further purified.

6. A method for obtaining a biologically active botulinum toxin type A, the method comprising the steps of:

(a) providing a fermentation medium which is free of an animal product;

(b) culturing a Clostridium botulinum type A bacterium in the fermentation medium under conditions which permit production of a botulinum toxin type A, and;

(c) recovering a biologically active botulinum toxin type A from the fermentation medium, wherein the fermentation medium comprises a protein product obtained from yeast or from a vegetable and wherein the vegetable is selected from the group consisting of soy, malt and corn.

7. A method for making a substantially animal product free pharmaceutical composition in which the active ingredient is a botulinum toxin, the method comprising the steps of:

(a) obtaining a biologically active botulinum toxin by;

(i) providing a fermentation medium which is free of an animal product;

(ii) culturing a Clostridium botulinum bacterium in the fermentation medium under conditions which permit production of a botulinum toxin, and;

(iii) recovering a biologically active botulinum toxin from the fermentation medium;

(b) formulating the botulinum toxin with a suitable excipient, thereby making a substantially animal product free pharmaceutical composition in which the active ingredient is a botulinum toxin,

wherein the fermentation medium comprises a protein product obtained from yeast or from a vegetable, and wherein the vegetable is selected from the group consisting of soy, malt and corn.

8. The method of claim 7, wherein the botulinum toxin is selected from the group consisting of botulinum toxins types A, B, C, D, E, F and G.

9. The method of claim 7, wherein the botulinum toxin is botulinum toxin type A.

10. The method of claim 7, wherein the botulinum toxin is further purified.

11. A method for making an animal product free pharmaceutical composition in which the active ingredient is a botulinum toxin type A, the method comprising the steps of:

(a) obtaining a biologically active botulinum toxin type A by;

(i) providing a fermentation medium which is free of an animal product;

(ii) culturing a Clostridium botulinum type A bacterium in the fermentation medium under conditions which permit production of a botulinum toxin type A, and;

(iii) recovering a biologically active botulinum toxin type A from the fermentation medium;

(b) formulating the botulinum toxin type A with a suitable excipient, thereby making an animal product free pharmaceutical composition in which the active ingredient is a botulinum toxin type A,

wherein the fermentation medium comprises a protein product obtained from yeast or from a vegetable, and wherein the vegetable is selected from the group consisting of soy, malt and corn.