Conotoxin polypeptide, preparation method and application thereof

A conotoxin and amino acid technology, applied in the preparation method of peptides, chemical instruments and methods, peptides, etc., can solve problems such as diplopia, unsuitable for wide use, unnatural expressions, etc., achieve inhibition of acetylcholine receptors, good economical and social effects, effects that apply to a wide range of

Inactive Publication Date: 2016-05-11
INFINITUS (CHINA) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, botulinum toxin is a neurotoxin protein produced by botulinum bacteria during reproduction, and the price is relatively high
Moreover, the injection of botulinum toxin has obvious side effects, such as headache, allergies, diplopia, and unnatural facial expressions, so it is not suitable for widespread use.

Method used

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  • Conotoxin polypeptide, preparation method and application thereof
  • Conotoxin polypeptide, preparation method and application thereof
  • Conotoxin polypeptide, preparation method and application thereof

Examples

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preparation example Construction

[0036] The present invention also provides a preparation method of the conotoxin polypeptide, comprising the following steps:

[0037] 1) Solid phase synthesis of HPA-amino resin;

[0038] 2) Cleavage of HPA-amino resin to obtain HPA crude peptide;

[0039] 3) The crude HPA peptide was dissolved in 1% acetic acid solution, separated by gel column chromatography, and purified by reverse-phase high-performance liquid chromatography to obtain the refined HPA peptide.

[0040] Wherein, step 1) of the preparation method of the present invention adopts the Fmoc / tBu synthesis strategy, uses the amino resin with a suitable degree of substitution as a carrier, and couples amino acids with Fmoc protecting groups one by one in solid phase synthesis to obtain HPA-amino resin.

[0041] Preferably, the solid-phase synthesis is Fmoc-Arg (Pbf)-OH and amino resin to react to synthesize Fmoc-Arg (Pbf)-amino resin, and then adopt other amino acids of coupling Fmoc protecting group to obtain HPA...

Embodiment 1

[0065] Example 1: Peptide Screening

[0066] Using bioinformatics methods, select the sequence of small peptides in the cysteine ​​ring of α3 / 5 conotoxin and perform homologous sequence alignment in blastp in NCBI (http: / / www.ncbi.nlm.nih.gov) , for one-to-one matching between proteins with the same sequence and the International Cosmetic Ingredient List (INCI), both of them can match the sequence as a candidate small peptide. Screen out NPA, HPA, WNPA, GKYYS, GKHFN.

Embodiment 2

[0067] Example 2: Synthesis of polypeptide HPA.

[0068] 1. Preparation of Fmoc-Arg(pbf)-amino resin

[0069] Add 10.03g of amino resin Fmoc-RinkAmide with a degree of substitution of 0.84mmol / g into a solid-phase reactor, add DMF to soak, make it fully swell for 30min, drain it, and add 20% piperidine / DMF solution to remove the Fmoc protecting group 2 The first reaction was 5 minutes, the second reaction was 15 minutes, and washed with DMF, DCM and methanol. Dissolve 11.1g of Fmoc-Arg(Pbf)-OH, 3.12g of HOBt, and 8.25g of HBTU in DMF and add to the reactor to balance for 5 minutes, add 7.0mL of DIPEA to react at room temperature for 1 hour, drain, and wash with DMF, DCM, and methanol for 3 times ,, 18.13g of Fmoc-Arg(pbf)-RinkAmide resin was obtained by dry weighing, and the detection degree of substitution was 0.68mmol / g.

[0070] 2. Preparation of HPA-amino resin

[0071] Put the Fmoc-Arg(pbf)-RinkAmide resin in the peptide bottle, add 20% piperidine / DMF solution to remov...

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Abstract

The invention relates to the field of polypeptide drugs, in particular to a conotoxin polypeptide, a preparation method and an application thereof, and especially relates to the conotoxin polypeptide acting on muscle-type nicotinic acetylcholine receptors. The amino acid sequence of the conotoxin polypeptide is HPA. The experimental results indicates that the conotoxin polypeptide HPA can act on the muscle-type nicotinic acetylcholine receptors to effectively inhibit the acetylcholine receptors, and can serve as antagonists of the muscle-type nicotinic acetylcholine receptors. The conotoxin polypeptide HPA can be used for preparing anti-wrinkle cosmetics, is broadly used and has good economic and social effects.

Description

technical field [0001] The invention relates to the field of polypeptide medicines, in particular to a conotoxin polypeptide and its preparation method and application, especially a conotoxin polypeptide acting on muscle nicotinic acetylcholine receptors. Background technique [0002] Muscle nicotinic acetylcholine receptors exist in the post-synaptic membrane at the neuromuscular junction and consist of five subunits, α2βγδ in fetal muscle and α2βεδ in adult muscle. When the nerve impulse is transmitted to the motor terminal, the presynaptic membrane releases acetylcholine, which acts on the α subunit of the post-synaptic membrane acetylcholine receptor, opens ion channels, and causes muscle contraction. [0003] BOTOX (Botox), which is widely used in the beauty industry, is botulinum toxin type A for injection. Its mechanism of action is to block the secretion of acetylcholine from nerve endings that can make muscles contract, so as to achieve the effect of paralyzing musc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/097C07K1/06C07K1/04A61K8/64A61Q19/00
CPCA61K8/64A61Q19/00C07K5/0821
Inventor 华洋林梁东李莉楠唐健
Owner INFINITUS (CHINA) CO LTD
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