Title:
RECOMBINANT BOTULINUM NEUROTOXIN OF TYPE A AND PREPARATION METHOD THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/226873
Kind Code:
A1
Abstract:
Provided are a recombinant BoNT/A and a preparation method thereof. The recombinant BoNT/A is a BoNT/A mutant, the BoNT/A mutant comprising a first peptide fragment and a second peptide fragment, which are linked through an interchain disulfide bond; wherein the first peptide fragment has a mutation at position 134 and/or position 165 compared to the light chain of a wild-type BoNT/A; and/or the second peptide fragment has at least one of the following mutation positions compared to the heavy chain of the wild-type BoNT/A: positions 791, 967, and 1060. The method includes: subjecting the first peptide fragment to a first renaturation treatment to obtain a first denatured product, subjecting the second peptide fragment to a second denaturation treatment to obtain a second denatured product, and subjecting the first denatured product and the second denatured product to a renaturation and assembly treatment to obtain the BoNT/A mutant.
Inventors:
SHEN YUBAO (CN)
BAO GUOQING (CN)
GUO LIMING (CN)
BAO GUOQING (CN)
GUO LIMING (CN)
Application Number:
PCT/CN2023/095060
Publication Date:
November 30, 2023
Filing Date:
May 18, 2023
Export Citation:
Assignee:
JHM BIOPHARMACEUTICAL HANGZHOU CO LTD (CN)
International Classes:
C07K14/33; A61K38/16; A61P17/00; A61P21/00; A61P21/02; A61P25/06; A61P27/02; A61P43/00; C12N1/21; C12N15/31; C12N15/70
Foreign References:
| CN114989271A | 2022-09-02 | |||
| CN115894641A | 2023-04-04 | |||
| US20220143157A1 | 2022-05-12 | |||
| CN103320459A | 2013-09-25 | |||
| CN113156105A | 2021-07-23 |
Other References:
SONG FU-YANG, MA CHEN-JIE, GAO SHAN, KANG LIN, WANG YU-JIONG, WANG JING-LIN: "Construction, Expression and Analysis of the Activity of Mutation of Recombinant Light Chain of Botulinum Neurotoxin Serotype A(BoNT/A LC)", LIFE SCIENCE RESEARCH, HUNAN SHIFAN DAXUE, SHENGMING KEXUE XUEYUAN,, CN, vol. 15, no. 03, 30 June 2011 (2011-06-30), CN , pages 263 - 267, XP009550627, ISSN: 1007-7847, DOI: 10.16605/j.cnki.1007-7847.2011.03.008
YAN LI, AOKI R, DOLLY J O: "Expression and characterisation of the heavy chain of tetanus toxin: Reconstitution of the fully-recombinant dichain protein in active form.", JOURNAL OF BIOCHEMISTRY, OXFORD UNIVERSITY PRESS, GB, vol. 125, no. 6, 1 June 1999 (1999-06-01), GB , pages 1200 - 1208, XP009016521, ISSN: 0021-924X
DATABASE UniProtKB 18 July 2018 (2018-07-18), ANONYMOUS : "RecName: Full=Botulinum neurotoxin type A", XP093111644, Database accession no. BXA1_CLOBH
YAN LI, AOKI R, DOLLY J O: "Expression and characterisation of the heavy chain of tetanus toxin: Reconstitution of the fully-recombinant dichain protein in active form.", JOURNAL OF BIOCHEMISTRY, OXFORD UNIVERSITY PRESS, GB, vol. 125, no. 6, 1 June 1999 (1999-06-01), GB , pages 1200 - 1208, XP009016521, ISSN: 0021-924X
DATABASE UniProtKB 18 July 2018 (2018-07-18), ANONYMOUS : "RecName: Full=Botulinum neurotoxin type A", XP093111644, Database accession no. BXA1_CLOBH
Attorney, Agent or Firm:
ZHIFAN & PARTNERS (CN)
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