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Botulinum toxin cell binding domain polypeptides and methods of use for skin rejuvenation

A botulinum toxin, binding domain technology for cosmetic applications

Pending Publication Date: 2021-02-09
ALLERGAN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Existing drug formulations contain full-length Clostridial toxin

Method used

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  • Botulinum toxin cell binding domain polypeptides and methods of use for skin rejuvenation
  • Botulinum toxin cell binding domain polypeptides and methods of use for skin rejuvenation
  • Botulinum toxin cell binding domain polypeptides and methods of use for skin rejuvenation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0319] Effects of Exemplary Polypeptides Provided According to Aspects of the Disclosure on Cellular Gene Expression in Normal Human Primary Fibroblasts

[0320] With a binding domain with BoNT / A (H C Treatment of normal human primary fibroblasts for 1, 2 or 3 days with polypeptides having substantially identical amino acid sequences to A) resulted in significant time-dependent changes in the expression of 10 fibroblast genes based on qPCR. Briefly, adult human dermal fibroblasts (HDFa) (cellResearchCorp PteLtd) were cultured in MEM medium containing 2% FBS for 2 weeks, and then treated with 1 μM of a polypeptide having SEQ ID NO: 19 for 1, 2 or 3 days. The time-dependent expression of genes known to be expressed in fibroblasts and involved in extracellular matrix (ECM) organization or epidermal self-renewal (keratinocyte stem cell factor) was evaluated. Total RNA was isolated using the RNAqueous kit from Ambion and cDNA was generated using Qiagen reagents following the manu...

Embodiment 2

[0322] Effects of the Exemplary Polypeptide of SEQ ID NO: 19 on Cellular Gene Expression in Normal Human Primary Fibroblasts

[0323] With 10nM, 100nM or 1μM with the binding domain of BoNT / A (H C / A) Treatment of normal human primary fibroblasts with polypeptides having substantially the same amino acid sequence for 1 day (24 hours) resulted in significant dose-dependent changes in the expression of 16 fibroblast-related genes based on qPCR. Briefly, adult human dermal fibroblasts (HDFa) (ThermoFisher Scientific, catalog number C0135C) were cultured in MEM medium containing 2% FBS for 2 weeks, and then treated with 10 nM, 100 nM or 1 μM of SEQ ID NO :19 peptide treatment continued for 1 day (24 hours). The expression of genes known to be expressed in fibroblasts and related to ECM organization, inflammation or epidermal self-renewal was evaluated. cDNA was generated by reverse transcription using the SUPERSCRIPT VILO cDNA Synthesis Kit (Thermo Scientific #11754050), then fu...

Embodiment 3

[0325] Effects of Exemplary Polypeptides Provided According to Aspects of the Disclosure on Cellular Gene Expression in Normal Human Primary Fibroblasts

[0326] With 1μM with the binding domain of BoNT / A (H C / A) a polypeptide having substantially the same amino acid sequence as the amino acid sequence or having the N-terminal half of the binding domain to BoNT (H CN / A) Treatment of normal human primary fibroblasts with polypeptides of substantially the same amino acid sequence resulted in similar significant changes in the expression of 16 fibroblast-related genes based on qPCR. Briefly, adult human dermal fibroblasts (HDFa) (ThermoFisher Scientific, catalog number C0135C) were cultured in MEM medium containing 2% FBS for 2 weeks, and then treated with 1 μM of the polypeptide having SEQ ID NO: 19 Or treatment with a polypeptide having SEQ ID NO: 21 for 1 day (24 hours). Expression of fibroblast-related genes known to be involved in ECM organization, inflammation, or epid...

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Abstract

A polypeptide having an amino acid sequence corresponding to a binding domain of a botulinum toxin is described. The polypeptide modulates expression of genes involved in, for example, collagen production and extra cellular matrix organization, and finds use, therefore in modulating skin quality attributes such as elasticity, firmness etc. Moreover, the polypeptide inhibits lipogenesis in specialized cells and finds use, therefore, in reducing skin oiliness, which is frequently observed in skin tissue afflicted with large pores and acne. Nucleic acids encoding the polypeptide, as well as vectors, host cells, and systems comprising the nucleic acids, are further described.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Application No. 62 / 608,119, filed December 20, 2017, and U.S. Provisional Application No. 62 / 727,640, filed September 6, 2018, the entire contents of which are incorporated herein by reference. [0003] sequence listing [0004] This application contains a Sequence Listing that has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy was created on December 17, 2018, named 19980-US-NTB_SL19980PROV2(NTB)_SL.txt, and was 71,994 bytes in size. technical field [0005] The present disclosure relates generally to polypeptides from the cell-binding domain of botulinum toxin and the use of said polypeptides in cosmetic applications related to skin regeneration. Background technique [0006] The anaerobic gram-positive bacterium Clostridium botulinum produces a potent polypeptide neurotoxin, botulinum toxin (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/33A61K38/48
CPCC12N9/52A61K38/4893C12Y304/24069A61P35/00A61P13/10A61P13/08A61P1/18A61P17/02A61P17/10A61P37/00A61P17/06A61P17/08A61P29/00C07K14/33Y02A50/30A61Q19/007A61Q19/08A61Q19/008A61K9/0014A61K9/0019C07K2319/00C07K14/315A61P43/00
Inventor B·P·杰克伊A·布里多安德森H·尤S·Z·马里克D·E·弗雷尔M·F·布林
Owner ALLERGAN INC
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