Stabilization of clostridium botulinum neurotoxin complex

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...

AI Technical Summary

Benefits of technology

[0016]The present 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. In particular, the present invention seeks to provide a method for producing a botulinum neurotoxin composition with improved stability properties in an efficient and economically advantageous manner.

[0022]In yet another embodiment, the composition comprises botulinum neurotoxin Type A and gamma-cyclodextrin. In this embodiment, the degradation of the neurotoxin at 4° C. was less than 2%, and the degradation of the neurotoxin at 25° C. was less than 55%, both over a 23-week period. Accordingly, the composition has an increased stability over time.

[0025]In accordance with another aspect of the invention, a method is provided for increasing the stability of botulinum neurotoxin comprising subjecting the botulinum neurotoxin to cyclodextrin to form the complex. Preferably, the molar ratio of the cyclodextrin to botulinum neurotoxin is at least 25:1 to 50:1, and most preferably 39:1. As used herein, the term “cyclodextrin” refers to a compound having a plurality of cyclopyranose units and include alpha, beta, gamma-cyclodextrins, any derivative or salt thereof and any combination thereof. The method of the invention provides a cyclodextrin-botulinum neurotoxin complex having markedly improved stability. Consequently, this improved stability serves to provide a botulinum neurotoxin with increased shelf-life. Preferably, the botulinum neurotoxin is subjected to the cyclodextrin in an aqueous system. The aqueous system has a pH of about 6.8 to 7.6 and can contain between 3 and 25 mM cyclodextrin, between 5 and 25 nM phosphate buffer. Most preferably, the aqueous system comprises 10 mM cyclodextrin, 10 mM sodium phosphate buffer, and has a pH 7.4.

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.

Images