Compositions of activated botulinum holotoxin type B (150 KD)
The present invention relates to pharmaceutical compositions of activated botulinum holotoxin type B (150 kD). In particular, the present invention relates to botulinum toxin type B pharmaceutical compositions wherein at least 75% of said botulinum toxin type B is activated (i.e., “nicked”), and wherein at least 75% said nicked botulinum toxin type B is a 150 kD holotoxin (i.e., “stripped”). The invention also relates to a process of activating and stripping botulinum toxin type B wherein at least 75% of said botulinum toxin type B is nicked, and wherein at least 75% of said nicked botulinum toxin type B is stripped. The invention further relates to methods for the treatment of a variety of neuromuscular diseases, pain, inflammatory and cutaneous disorders comprising administering a pharmaceutical composition of activated botulinum holotoxin type B (150 kD) wherein at least 75% of said botulinum toxin type B is nicked, and wherein at least 75% of said nicked botulinum toxin type B is stripped. The invention also relates to a process of stripping botulinum toxin type B, comprising the stages of: cell growth, activation, purification, and dilution; wherein at least one dissociating reagent is administered to a volume of said botulinum toxin type B; and wherein said dissociating reagent administered increases the levels of stripped botulinum toxin type B to at least 75%.
1 . A pharmaceutical composition comprising:
(a) activated botulinum toxin type B; and
(b) at least one excipient;
wherein at least 75% of said botulinum toxin type B is nicked; and
wherein at least 75% of said nicked botulinum toxin type B is an approximately 150 kD holotoxin.
2 . The pharmaceutical composition of claim 1 , wherein greater than 75 percent of said botulinum toxin B is nicked.
3 . The pharmaceutical composition of claim 1 , wherein approximately about 75 percent to about 85 percent of said botulinum toxin type B is nicked.
4 . The pharmaceutical composition of claim 1 , wherein greater than 80 percent of said botulinum toxin B is nicked.
5 . The pharmaceutical composition of claim 1 , wherein greater than 85 percent of said botulinum toxin B is nicked.
6 . The pharmaceutical composition of claim 1 , wherein approximately about 85 percent to about 95 percent of said botulinum toxin type B is nicked.
7 . The pharmaceutical composition of claim 1 , wherein greater than 90 percent of said botulinum toxin B is nicked.
8 . The pharmaceutical composition of claim 1 , wherein approximately about 95 percent to about 100 percent of said botulinum toxin type B is nicked.
9 . The pharmaceutical composition of claim 1 , wherein greater than 95 percent of said botulinum toxin B is nicked.
10 . The pharmaceutical composition of claim 1 , wherein greater than 99 percent of said botulinum toxin B is nicked.
11 . The pharmaceutical composition of claim 1 , wherein greater than 75 percent of said botulinum toxin B is stripped.
12 . The pharmaceutical composition of claim 1 , wherein approximately about 75 percent to about 85 percent of said botulinum toxin type B is stripped.
13 . The pharmaceutical composition of claim 1 , wherein greater than 80 percent of said botulinum toxin B is stripped.
14 . The pharmaceutical composition of claim 1 , wherein greater than 85 percent of said botulinum toxin B is stripped.
15 . The pharmaceutical composition of claim 1 , wherein approximately about 85 percent to about 95 percent of said botulinum toxin type B is stripped.
16 . The pharmaceutical composition of claim 1 , wherein greater than 90 percent of said botulinum toxin B is stripped.
17 . The pharmaceutical composition of claim 1 , wherein approximately about 95 percent to about 100 percent of said botulinum toxin type B is stripped.
18 . The pharmaceutical composition of claim 1 , wherein greater than 95 percent of said botulinum toxin B is stripped.
19 . The pharmaceutical composition of claim 1 , wherein greater than 99 percent of said botulinum toxin B is stripped.
20 . The pharmaceutical composition of claim 1 , wherein said at least one excipient is selected from the group consisting of: buffers, carriers, stabilizers, preservatives, diluents, vehicles, bulking agents, albumins, gelatins, collagens, proteins, polysaccharides, metals, non-oxidizing amino acid derivatives, and sodium chloride.
21 . The pharmaceutical composition of claim 1 , wherein said at least one excipient is albumin.
22 . The pharmaceutical composition of claim 1 , wherein said at least one excipient is a buffer.
23 . The pharmaceutical composition of claim 7 , wherein said buffer is a succinate buffer.
24 . A process of activating and stripping botulinum toxin type B, comprising the stages of:
cell growth, activation, purification, and dilution; wherein at least one protease is administered to a volume of said botulinum toxin type B; wherein said protease administered increases the levels of nicked botulinum toxin type B to at least 75%;
wherein at least one dissociating reagent is administered to a volume of said nicked botulinum toxin type B; and wherein said dissociating reagent administered increases the levels of stripped botulinum toxin type B to at least 75%.
25 . The process of claim 24 , wherein said at least one protease administered is selected from the group consisting of: trypsin, immobilized TPCK-trypsin, metalloproteases, endogenous proteases, bacterial proteases, and gastric proteases.
26 . The process of claim 25 , wherein said protease administered is trypsin.
27 . The process of claim 24 , wherein said at least one dissociating reagent administered is a succinate buffer.
28 . The process of claim 24 , wherein greater than 75 percent of said botulinum toxin B is nicked.
29 . The process of claim 24 , wherein approximately about 75 percent to about 85 percent of said botulinum toxin type B is nicked.
30 . The process of claim 24 , wherein greater than 80 percent of said botulinum toxin B is nicked.
31 . The process of claim 24 , wherein greater than 85 percent of said botulinum toxin B is nicked.
32 . The process of claim 24 , wherein approximately about 85 percent to about 95 percent of said botulinum toxin type B is nicked.
33 . The process of claim 24 , wherein greater than 90 percent of said botulinum toxin B is nicked.
34 . The process of claim 24 , wherein approximately about 95 percent to about 100 percent of said botulinum toxin type B is nicked.
35 . The process of claim 24 , wherein greater than 95 percent of said botulinum toxin B is nicked.
36 . The process of claim 24 , wherein greater than 99 percent of said botulinum toxin B is nicked.
37 . The process of claim 24 , wherein greater than 75 percent of said botulinum toxin B is stripped.
38 . The process of claim 24 , wherein approximately about 75 percent to about 85 percent of said botulinum toxin type B is stripped.
39 . The process of claim 24 , wherein greater than 80 percent of said botulinum toxin B is stripped.
40 . The process of claim 24 , wherein greater than 85 percent of said botulinum toxin B is stripped.
41 . The process of claim 24 , wherein approximately about 85 percent to about 95 percent of said botulinum toxin type B is stripped.
42 . The process of claim 24 , wherein greater than 90 percent of said botulinum toxin B is stripped.
43 . The process of claim 24 , wherein approximately about 95 percent to about 100 percent of said botulinum toxin type B is stripped.
44 . The process of claim 24 , wherein greater than 95 percent of said botulinum toxin B is stripped.
45 . The process of claim 24 , wherein greater than 99 percent of said botulinum toxin B is stripped.
46 . A method of treating a disorder, comprising administering to a patient in need thereof, a pharmaceutical composition comprising:
(a) activated botulinum toxin type B; and
(b) at least one excipient;
wherein at least 75% of said botulinum toxin type B is nicked; and
wherein at least 75% of said nicked botulinum toxin type B is stripped.
47 . The method of claim 46 , wherein greater than 75 percent of said botulinum toxin B in said pharmaceutical composition is nicked.
48 . The method of claim 46 , wherein approximately about 75 percent to about 85 percent of said botulinum toxin type B in said pharmaceutical composition is nicked.
49 . The method of claim 46 , wherein greater than 80 percent of said botulinum toxin B in said pharmaceutical composition is nicked.
50 . The method of claim 46 , wherein greater than 85 percent of said botulinum toxin B in said pharmaceutical composition is nicked.
51 . The method of claim 46 , wherein approximately about 85 percent to about 95 percent of said botulinum toxin type B in said pharmaceutical composition is nicked.
52 . The method of claim 46 , wherein greater than 90 percent of said botulinum toxin B in said pharmaceutical composition is nicked.
53 . The method of claim 46 , wherein approximately about 95 percent to about 100 percent of said botulinum toxin type B in said pharmaceutical composition is nicked.
54 . The method of claim 46 , wherein greater than 95 percent of said botulinum toxin B in said pharmaceutical composition is nicked.
55 . The method of claim 46 , wherein greater than 99 percent of said botulinum toxin B in said pharmaceutical composition is nicked.
56 . The method of claim 46 , wherein greater than 75 percent of said botulinum toxin B in said pharmaceutical composition is stripped.
57 . The method of claim 46 , wherein approximately about 75 percent to about 85 percent of said botulinum toxin type B in said pharmaceutical composition is stripped.
58 . The method of claim 46 , wherein greater than 80 percent of said botulinum toxin B in said pharmaceutical composition is stripped.
59 . The method of claim 46 , wherein greater than 85 percent of said botulinum toxin B in said pharmaceutical composition is stripped.
60 . The method of claim 46 , wherein approximately about 85 percent to about 95 percent of said botulinum toxin type B in said pharmaceutical composition is stripped.
61 . The method of claim 46 , wherein greater than 90 percent of said botulinum toxin B in said pharmaceutical. composition is stripped.
62 . The method of claim 46 , wherein approximately about 95 percent to about 100 percent of said botulinum toxin type B in said pharmaceutical composition is stripped.
63 . The method of claim 46 , wherein greater than 95 percent of said botulinum toxin B in said pharmaceutical composition is stripped.
64 . The method of claim 46 , wherein greater than 99 percent of said botulinum toxin B in said pharmaceutical composition is stripped.
65 . The method of claim 46 , wherein said at least one excipient of said pharmaceutical composition is selected from the group consisting of: buffers, carriers, stabilizers, preservatives, diluents, vehicles, bulking agents, albumins, gelatins, collagens, proteins, polysaccharides, metals, non-oxidizing amino acid derivatives, and sodium chloride.
66 . The method of claim 65 , wherein said at least one excipient of said pharmaceutical composition is albumin.
67 . The method of claim 65 , wherein said at least one excipient of said pharmaceutical composition is a buffer.
68 . The method of claim 67 , wherein said buffer is a succinate buffer.
69 . The method of claim 46 , wherein said disorder is selected from the group consisting of:
ophthalmologic disorders, neuromuscular diseases, otorhinolaryngological disorders, urogenital disorders, dermatological disorders, pain disorders, inflammatory disorders, secretory disorders, and cutaneous disorders or cosmetic treatment.
70 . A process of stripping botulinum toxin type B, comprising the stages of: cell growth, activation, purification, and dilution; wherein at least one dissociating reagent is administered to a volume of said botulinum toxin type B; and wherein said dissociating reagent administered increases the levels of stripped botulinum toxin type B to at least 75%.
71 . The process of claim 70 , wherein greater than 75 percent of said botulinum toxin B is stripped.
72 . The process of claim 70 , wherein approximately about 75 percent to about 85 percent of said botulinum toxin type B is stripped.
73 . The process of claim 70 , wherein greater than 80 percent of said botulinum toxin B is stripped.
74 . The process of claim 70 , wherein greater than 85 percent of said botulinum toxin B is stripped.
75 . The process of claim 70 , wherein approximately about 85 percent to about 95 percent of said botulinum toxin type B is stripped.
76 . The process of claim 70 , wherein greater than 90 percent of said botulinum toxin B is stripped.
77 . The process of claim 70 , wherein approximately about 95 percent to about 100 percent of said botulinum toxin type B is stripped.
78 . The process of claim 70 , wherein greater than 95 percent of said botulinum toxin B is stripped.
79 . The process of claim 70 , wherein greater than 99 percent of said botulinum toxin B is stripped.