WO2021243756A1 - 杨梅素抑制新型冠状病毒的药物应用 - Google Patents

杨梅素抑制新型冠状病毒的药物应用 Download PDF

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WO2021243756A1
WO2021243756A1 PCT/CN2020/096824 CN2020096824W WO2021243756A1 WO 2021243756 A1 WO2021243756 A1 WO 2021243756A1 CN 2020096824 W CN2020096824 W CN 2020096824W WO 2021243756 A1 WO2021243756 A1 WO 2021243756A1
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myricetin
ncov
novel coronavirus
inhibition
protease
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PCT/CN2020/096824
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French (fr)
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宋昆元
陈伟伟
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上海爱启医药技术有限公司
宋昆元
陈伟伟
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Priority to US17/287,417 priority Critical patent/US20220040142A1/en
Publication of WO2021243756A1 publication Critical patent/WO2021243756A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 

Definitions

  • the invention belongs to the technical field of medicine, and specifically relates to a drug application of myricetin for inhibiting a new type of coronavirus.
  • New coronary pneumonia has broken out worldwide since 2020 and may coexist with humans for a long time. There is currently no approved specific medicine available.
  • 3-chymotrypsin-like protease (3-chymotrypsin-like protease), the main protease (M pro , also known as 3CLpro), is encoded by ORF1 (located in nsp5), located in the central region of the replicase gene, and is a new type of coronavirus A key protein in RNA replication. The mechanism of action is: after the new coronavirus invades the cell, it will use the host cell to synthesize two ultra-long replicase polypeptides (pp1a and pp1ab) necessary for self-replication.
  • ORF1 located in nsp5
  • pp1a and pp1ab two ultra-long replicase polypeptides
  • the replicase polypeptide needs to be further cut into multiple proteins (such as RdRp, helicase, etc.), and then assembled into the replication and transcription machinery required for the virus to initiate the replication of its own genetic material.
  • M pro has at least 11 cleavage sites on the replicase polypeptide. Only when these sites on the replicase polypeptide are normally cut, they are assembled into a replication transcription machine to initiate virus replication. Given that the M pro protease is very important in the virus replication process, and there is no similar protein in the human body, the main protease M pro has become a potential key drug target against the new coronavirus.
  • the purpose of the present invention is to provide a medicine capable of inhibiting the novel coronavirus (2019-nCoV): myricetin.
  • the present invention provides an application of myricetin in inhibiting the novel coronavirus (2019-nCoV).
  • it may also have the characteristic that myricetin can bind to the 2019-nCoV-M pro protease, thereby inhibiting the 2019-nCoV virus.
  • Fig. 1 is a calculation diagram of the IC50 value of myricetin against the target M pro of the novel coronavirus (2019-nCoV) in the experimental example of the present invention.
  • the myricetin involved in the present invention can bind to the 2019-nCoV-Mpro protease, and its IC50 value for the target M pro of the new coronavirus (2019-nCoV) is 0.582 ⁇ 0.0912 ⁇ M, which has a significant effect and can inhibit the 2019-nCoV virus .
  • Myricetin English name: Myricetin; Chinese chemical name: 3,5,7-trihydroxy-2-(3,4,5-trihydroxyphenyl)-4H-1-benzofuran-4-one; English chemistry Name: 3,3',4',5,5',7-Hexahydroxyflavone; CAS Number: 529-44-2; Molecular Formula: C 15 H 10 O 8 ; Molecular Weight: 318.24; Chemical Structure:
  • Myricetin has the effect of inhibiting the new coronavirus (2019-nCoV).
  • This example is the detection of M pro protease activity inhibition of the targeted 2019-nCoV virus.
  • the fluorescence resonance energy transfer method was used to evaluate and determine the inhibitory activity of myricetin on 2019-nCoV-Mpro protease.
  • the volume of the entire enzymatic reaction system is 120 ⁇ L, the final concentration of protease is 30nM, and the final concentration of substrate is 20 ⁇ M.
  • the buffer of the reaction system includes 50mM Tris pH7.3, 1mM EDTA.
  • the excitation light and emission light are 340nm and 405nm, respectively.
  • the test time is 10min, and the fluorescence value is read every 30s.
  • the final result takes the reading of the first 2 minutes to fit the reaction rate, and compares it with the control group (DMSO) to calculate the inhibition rate.
  • DMSO control group
  • myricetin has an IC50 value of 0.582 ⁇ 0.0912 ⁇ M for the target M pro of the new coronavirus (2019-nCoV), which has a significant effect.

Abstract

一种杨梅素抑制新型冠状病毒的药物应用，研究证明杨梅素能结合2019-nCoV-Mpro蛋白酶，其针对新型冠状病毒(2019-nCoV)的靶点M pro的IC50值为0.582±0.0912μM，具有显著效果，能够抑制2019-nCoV病毒。

Description

杨梅素抑制新型冠状病毒的药物应用 技术领域
本发明属于医药技术领域，具体涉及一种杨梅素抑制新型冠状病毒的药物应用。
背景技术
新冠肺炎自2020年世界范围内爆发，并且可能和人类长期共存，目前无批准的特效药可用。
3-胰凝乳蛋白酶样蛋白酶(3-chymotrypsin-like protease)，即主要蛋白酶(M pro，也称为3CLpro)，由ORF1编码(定位于nsp5)，位于复制酶基因中心区域，是新型冠状病毒RNA复制时的一个关键蛋白质。其作用机制为：新冠病毒入侵细胞后，会利用宿主细胞合成自身复制必需的两条超长复制酶多肽(pp1a和pp1ab)。复制酶多肽需进一步被剪切成多个蛋白(如RdRp、helicase等)，进一步组装成用于病毒启动自身遗传物质复制所需的复制转录机器。M pro在复制酶多肽上存在至少11个切割位点，只有当复制酶多肽上这些位点被正常切割后，组装成复制转录机器，启动病毒复制。鉴于M pro蛋白酶在病毒复制过程中至关重要，且人体中并无类似的蛋白质，因此主蛋白酶M pro成为一个抗新冠病毒的潜在关键药靶。
发明内容
本发明目的是提供一种能够抑制新型冠状病毒(2019-nCoV)的药物：杨梅素。
本发明提供了一种杨梅素在抑制新型冠状病毒(2019-nCoV)方 面的应用。
进一步，在本发明提供的应用中，还可以具有这样的特征：杨梅素能结合2019-nCoV-M pro蛋白酶，从而抑制2019-nCoV病毒。
附图说明
图1是本发明的实验例中杨梅素对新型冠状病毒(2019-nCoV)的靶点M pro的的IC50值的计算图。
本发明提供了如下优点：
本发明所涉及的杨梅素能结合2019-nCoV-Mpro蛋白酶，其针对新型冠状病毒(2019-nCoV)的靶点M pro的IC50值为0.582±0.0912μM，具有显著效果，能够抑制2019-nCoV病毒。
具体实施方式
为了使本发明实现的技术手段、创作特征、达成目的与功效易于明白了解，以下结合实施例对本发明的杨梅素在抑制新型冠状病毒方面的药物应用作具体阐述。
杨梅素：英文名称：Myricetin；中文化学名：3,5,7-三羟基-2-(3,4,5-三羟基苯基)-4H-1-苯并呋喃-4-酮；英文化学名：3,3',4',5,5',7-Hexahydroxyflavone；CAS号：529-44-2；分子式：C 15H 10O 8；分子量：318.24；化学结构：
Figure PCTCN2020096824-appb-000001
杨梅素具有抑制新型冠状病毒(2019-nCoV)的效果。
<实验实施例>
本实施例为靶向2019-nCoV病毒M pro蛋白酶活性抑制检测。
利用荧光共振能量转移方法评价测定杨梅素对2019-nCoV-Mpro蛋白酶的抑制活性。
检测方法：整个酶促反应体系的体积为120μL，蛋白酶的终浓度为30nM，底物终浓度为20μM。反应体系的缓冲液包括50mM Tris pH7.3、1mM EDTA。在96孔板中加入2019-nCoV-Mpro蛋白酶和不同浓度的杨梅素，30℃孵育10min，加入底物并迅速放入酶标仪中读数。激发光和发射光分别为340nm和405nm。测试时间为10min，每隔30s读一次荧光值。最终结果取前2min的读值拟合反应速率，并与对照组(DMSO)比较，计算抑制率。以Graghpad-prism5.0作图，如图1，计算杨梅素对应时间点IC50值。
检测结果如表1所示：
表1.杨梅素靶向2019-nCoV-Mpro蛋白酶活性测试结果
样品 IC50(μg/ml) IC50(μM)
杨梅素 0.185±0.029μg/ml 0.582±0.0912μM
从表1中可以看出杨梅素针对新型冠状病毒(2019-nCoV)的靶点M pro的IC50值为0.582±0.0912μM，具有显著效果。
本发明所涉及的杨梅素抑制新型冠状病毒的药物应用并不限于具体实施例的范围。以上内容仅为本发明的基本说明，而依据本发明 的技术方案所作的任何等效变换，均应属于本发明的保护范围。

Claims (2)

  1. 杨梅素在抑制新型冠状病毒(2019-nCoV)方面的应用。
  2. 根据权利要求1所述的应用，其特征在于：杨梅素能结合2019-nCoV-M pro蛋白酶，从而抑制2019-nCoV病毒。
PCT/CN2020/096824 2020-06-03 2020-06-18 杨梅素抑制新型冠状病毒的药物应用 WO2021243756A1 (zh)

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CN112168899B (zh) * 2020-09-30 2022-07-01 上海中医药大学 抑制冠状病毒3cl蛋白水解酶的藤茶提取物及其用途
CN112546038A (zh) * 2020-11-19 2021-03-26 澳门科技大学 杨梅素在制备预防或治疗冠状病毒、流感病毒的药物中的应用
EP4011367A1 (en) * 2020-12-09 2022-06-15 Dompe' Farmaceutici S.P.A. Compounds for use in the treatment of covid-19
CN114983993B (zh) * 2021-03-02 2023-11-17 中国科学院上海药物研究所 杨梅素和二氢杨梅素磷酸酯类化合物在防治新冠肺炎药物中的应用

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