Harvard College

Harvard Corporation

Zhongshan University

Lisheng Peng

Min Dong

Pål Erik Gustav Stenmark

Ronnie Per Arne Berntsson

Liang Tao

Wenyan Wu

Wenli Yang

Anlong Xu

Xiaofen Zhong

Jianwen Wei

Ronnie Per-Arne Berntsson

Disclosed herein is a botulinum neurotoxin (BoNT) polypeptide with a modified receptor binding domain (HC) having one or more amino acid mutations that modify the binding of the BoNT to the receptor. Specific mutations and combinations of mutations are also disclosed. Isolated modified HC, polypeptides comprising the modified HC, chimeric molecules, pharmaceutical compositions, and methods of making and using the same are also disclosed. Methods of identifying additional such modified receptor binding domains, are further disclosed.

Engineered botulinum neurotoxin

Disclosed herein are botulinum neurotoxin (BoNT) polypeptides with a modified receptor binding domain ofserotype B (B-H), comprising one or more substitution mutations corresponding to substitution mutations in serotype B, strain 1, V1118M; Y1183M; E1191M; E1191I; E1191Q; E1191T; S1199Y; S1199F; S1199L; S1201V; or combinations thereof. Specific combination mutations include E1191M and S1199L, E1191M and S1199Y, E1191M and S1199F, E1191Q and S1199L, E1191Q and S1199Y, or E1191Q and S1199F. Other substitution mutations are also disclosed. Isolated modified receptor binding domains, chimeric molecules, pharmaceutical compositions, and methods of using the same are also disclosed.

Disclosed herein areneurotoxin (BoNT) polypeptides with a modified receptor binding domain ofserotype B (B-H), comprising one or more substitution mutations corresponding to substitution mutations in serotype B, strain 1, V1118M; Y1183M; E1191M; E1191I; E1191Q; E1191T; S1199Y; S1199F; S1199L; SI 20 IV; or combinations thereof. Specific combination mutations include E1 191M and S1199L, E1191M and S1199Y, E1191M and S1199F, E1191Q and S1199L, E1191Q and S 1199Y, or E 1191 Q and S 1199F. Other substitution mutations are also disclosed. Isolated modified receptor binding domains, chimeric molecules, pharmaceutical compositions, and methods of using the same are also disclosed.

In this invention, three genes encoding short chain neurotoxin are obtained by sea snake&#x2014;venom gland cDNA library construction and DNA sequencing, named sn12, sn36, and sn160, respectively. The three genes are modified and amplified by PCR method, then cloned into expression vector PETTRX. The three neurotoxin genes are all highly expressed in soluble fusion protein instain BL21-DE3 which is used as a expression host when induced with IPTG. Studies on the analgesic effects of the three recombinant short chain neurotoxins showed that three proteins, SN12, SN36 and SN160 all can significantly increase the pain threshold value of mouse.

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Growing community of inventors

Guangzhou, China

Lisheng Peng