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Stabilization of clostridium botulinum neurotoxin complex

a neurotoxin and complex technology, applied in the field of stabilization of clostridium botulinum neurotoxin, can solve the problems of food-borne botulism, short shelf life of the composition, improper storage, etc., and achieve the effects of improving stability properties, and reducing the risk of foodborne botulism

Inactive Publication Date: 2010-09-02
B B SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The cyclodextrin-botulinum neurotoxin complex maintains potency for at least 23 weeks, with less than 7% degradation at 4°C and less than 65% at 25°C, significantly increasing the stability and shelf life of the neurotoxin.

Problems solved by technology

Food-borne botulism results from the consumption of improperly stored foods in which anaerobic C. botulinum grows and releases the toxin.
One major drawback to using the commercially available botulinum toxin preparations is the very short shelf life of the composition.
In this regard, the actual usage of the pharmaceutical composition should be administered within about four hours after reconstitution because the botulinum toxin is very susceptible to denaturation due to surface denaturaion, heat, and alkaline conditions.
In fact, it has been thought that cyclic polymers including cyclodextrins can not be used to preserve or stabilize a botulinum neurotoxin.

Method used

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  • Stabilization of clostridium botulinum neurotoxin complex
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  • Stabilization of clostridium botulinum neurotoxin complex

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Experimental program
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example

Preparation of C. botulinum Type A

[0059]The C. botulinum Type A (strain Hall) complex was prepared by the method described in Cai et al. Enhancement of the Endopeptidase Activity of Botulinum Neurotoxin by Its Associated Proteins and Dithiothreitol, Biochemistry, 1999, 38, 6903-6910, the entire contents of which are incorporated herein by reference. The purified Type A complex was subjected to a buffer exchange using a 5 mL Sephadex G-25 column equilibrated with 10 column volumes of 10 mM sodium phosphate, pH 7.4. The Type A complex was determined to have a typical subunit makeup by SDS-PAGE analysis. The Type A complex, at physiological pH of 7.4, was diluted to a 0.75 mg / mL concentration and 1 mL aliquots were placed into eight 1.5 mL microcentrifuge tubes.

[0060]1.5 mg mL−1 solutions of the Type A Botulinum neurotoxin complex at both 25° C. and 4° C. were analyzed by high performance liquid chromatography using a gel filtration (size exclusion) column (HPLC-GF) both in the presenc...

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Abstract

A stable composition including a non-covalent complex of a botulinum neurotoxin and a cyclodextrin and a method of preserving botulinum neurotoxin and for producing a botulinum neurotoxin composition with improved stability properties in an efficient and economically advantageous manner. The invention seeks to alleviate the problems associated with rapid degradation or denaturation of botulinum neurotoxin by providing a novel composition that exhibits improved stability properties. The botulinum neurotoxin is preferably stabilized by forming a cyclodextrin complex.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a divisional application of Ser. No. 11 / 525,566 filed Sep. 21, 2006, which claims the benefit of provisional patent application No. 60 / 720,854 filed Sep. 26, 2005, the entire content of each of which is expressly incorporated herein by reference thereto.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The present invention relates to a pharmaceutical composition comprising Clostridium botulinum neurotoxin and to a method of stabilizing the same. Particularly, the present invention is directed to a stabilized pharmaceutical composition including C. botulinum Type A and a cyclodextrin.[0004]2. Description of Related Art[0005]Botulinum neurotoxins are produced from anaerobic bacillus Clostridium botulinum. Seven related protein neurotoxins, known as serotypes A through G, are produced by different strains of the bacillus. Each of the seven serotypes of the botulinum neurotoxins is a large protein having a m...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/16C07K14/33A61P21/00
CPCC12Y304/24069C12N9/6416A61P21/00
Inventor SINGH, BAL RAMRESS, ANDREW M.
Owner B B SCI
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