药学学报

Process of virus infection	Potential target	Potential effect of drug
Virus adsorption	ACE2, S-ACE2	Interference the recognition for ACE2 receptor and suppress virus invasion, therefore they can be used for COVID prevention
Membrane fusion	TMPRSS2	Inhibit the fusion between membrane protein of virus and cell membrane, protect cells against SARS-CoV-2 induced COVID-19
Non-structural protein	NSP1, NSP3, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12 (RNA dependent RNA polymerase, RdRp), NSP13 (RNA helicase), NSP15, NSP16	Inhibit the activity of non-structural functional proteins and interference the procedure of virus replication. Used for COVID-19 to inhibit virus replication
Virus replication	PLpro, 3CLpro, Furin	Inhibit the activity of different enzymes in virus replication process
Pathogenic factors	IL-6, IL-1β, TNF-α	Anti-inflammation, immunity regulation

Process of virus infection	Potential target	Potential effect of drug
Virus adsorption	ACE2, S-ACE2	Interference the recognition for ACE2 receptor and suppress virus invasion, therefore they can be used for COVID prevention
Membrane fusion	TMPRSS2	Inhibit the fusion between membrane protein of virus and cell membrane, protect cells against SARS-CoV-2 induced COVID-19
Non-structural protein	NSP1, NSP3, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12 (RNA dependent RNA polymerase, RdRp), NSP13 (RNA helicase), NSP15, NSP16	Inhibit the activity of non-structural functional proteins and interference the procedure of virus replication. Used for COVID-19 to inhibit virus replication
Virus replication	PLpro, 3CLpro, Furin	Inhibit the activity of different enzymes in virus replication process
Pathogenic factors	IL-6, IL-1β, TNF-α	Anti-inflammation, immunity regulation

English name	CAS	Source
Hesperidin	520-26-3	Galium aparine, Citrus medica var. sarcodactylis, Citrus chachiensis, Poncirus trifoliata, Capsella bursa-pastoris, Citrus tankan, Schizonepeta tenuifolia [Syn. Nepeta tenuifolia], Citrus reticulata, Citrus medica, Citrus limonia, Microtoena prainiana, Citrus limon, Mentha longifolia, Galium mollugo, Citrus unshiu, Citrus aurantium
Isosakuranetin-7-rutinoside		Citrus sinensis
Narirutin	14259-46-2	Citrus sinensis, Citrus sp.
Diosmin	520-27-4	Galium aparine, Sophora viciifolia, Toddalia asiatica [Syn. Toddalia aculeata; Paullinia asiatica], Citrus medica var. sarcodactylis, Scabiosa caucasica, Capsella bursa-pastoris, Scrophularia nodosa, Rosmarinus officinalis, Citrus limon, Urtica cannabina, Zanthoxylum nitidum, Zanthoxylum avicennae
Neohesperidin	13241-33-3	Poncirus trifoliata, Citrus limon, Citrus unshiu, Citrus aurantium
Chrysoeriol 7-O-rutinoside		Saussurea medusa
Acaciin	480-36-4	Chrysanthemum boreale, Robinia pseudoacacia, Trapa bispinosa, Agastache rugosus, Buddleja officinalis, Chrysanthemum indicum
Ombuoside	20188-85-6	Phytolacca dioica, Gynostemma pentaphyllum, Flyriella parryi
Uralsaponin A	103000-77-7	Glycyrrhiza uralensis, Glycyrrhiza kansuensis
Licoricesaponin H2	135815-61-1	Glycyrrhiza uralensis, Glycyrrhiza inflata
Glycyrrhizic acid	1405-86-3	Glycyrrhiza aspera, Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Abrus precatorius, Glycyrrhiza inflata
Licoricesaponin G2	118441-84-2	Glycyrrhiza uralensis, Glycyrrhiza inflata
Glyeurysaponin	137476-70-1	Glycyrrhiza kansuensis
Araboglycyrrhizin	121687-83-0	Glycyrrhiza inflata
Uralsaponin B	105038-43-5	Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Glycyrrhiza inflata
Licoricesaponin J2	134250-13-8	Glycyrrhiza uralensis
Baicalin	21967-41-9	Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Salvia miltiorrhiza, Scutellaria amoena, Scutellaria rehderiana, Scutellaria baical ensis, Oroxylum indicum, Scutellaria viscidula
Baicalein-7-O-beta-D- glucopyranoside	57396-78-8	Scutellaria baicalensis, Oroxylum indicum
Chrysin 7-O-beta-galactopyranuronoside		Centaurea pseudoscabiosa ssp. pseudoscabiosa
Baicalein-6-glucuronide	35990-03-5	Oroxylum indicum
5, 6, 4'-Trihydroxyflavone-7- O-beta-D-galactonic acid		Erigeron breviscapus
Breviscapine	116122-36-2	Erigeron breviscapus
Scutellarin	27740-01-8	Scutellaria barbata [Syn. Scutellaria rivularis], Clerodendron fragrans, Plantago major, Erigeron breviscapus, Sorbaria arborea, Scutellaria altissima, Scutellaria baicalensis, Perilla frutescens var. acuta [Syn. Perilla frutescens var. purpurascens], Oroxylum indicum, Sorbaria sorbifolia
Oroxylin A 7-O-glucuronide	36948-76-2	Scutellaria baicalensis
Baicalein	491-67-8	Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Scutellaria amoena, Scutellaria rehderiana, Scutellaria baicalensis, Scutellaria likiangensis, Oroxylum indicum, Scutellaria viscidula

English name	CAS	Source
Hesperidin	520-26-3	Galium aparine, Citrus medica var. sarcodactylis, Citrus chachiensis, Poncirus trifoliata, Capsella bursa-pastoris, Citrus tankan, Schizonepeta tenuifolia [Syn. Nepeta tenuifolia], Citrus reticulata, Citrus medica, Citrus limonia, Microtoena prainiana, Citrus limon, Mentha longifolia, Galium mollugo, Citrus unshiu, Citrus aurantium
Isosakuranetin-7-rutinoside		Citrus sinensis
Narirutin	14259-46-2	Citrus sinensis, Citrus sp.
Diosmin	520-27-4	Galium aparine, Sophora viciifolia, Toddalia asiatica [Syn. Toddalia aculeata; Paullinia asiatica], Citrus medica var. sarcodactylis, Scabiosa caucasica, Capsella bursa-pastoris, Scrophularia nodosa, Rosmarinus officinalis, Citrus limon, Urtica cannabina, Zanthoxylum nitidum, Zanthoxylum avicennae
Neohesperidin	13241-33-3	Poncirus trifoliata, Citrus limon, Citrus unshiu, Citrus aurantium
Chrysoeriol 7-O-rutinoside		Saussurea medusa
Acaciin	480-36-4	Chrysanthemum boreale, Robinia pseudoacacia, Trapa bispinosa, Agastache rugosus, Buddleja officinalis, Chrysanthemum indicum
Ombuoside	20188-85-6	Phytolacca dioica, Gynostemma pentaphyllum, Flyriella parryi
Uralsaponin A	103000-77-7	Glycyrrhiza uralensis, Glycyrrhiza kansuensis
Licoricesaponin H2	135815-61-1	Glycyrrhiza uralensis, Glycyrrhiza inflata
Glycyrrhizic acid	1405-86-3	Glycyrrhiza aspera, Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Abrus precatorius, Glycyrrhiza inflata
Licoricesaponin G2	118441-84-2	Glycyrrhiza uralensis, Glycyrrhiza inflata
Glyeurysaponin	137476-70-1	Glycyrrhiza kansuensis
Araboglycyrrhizin	121687-83-0	Glycyrrhiza inflata
Uralsaponin B	105038-43-5	Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Glycyrrhiza inflata
Licoricesaponin J2	134250-13-8	Glycyrrhiza uralensis
Baicalin	21967-41-9	Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Salvia miltiorrhiza, Scutellaria amoena, Scutellaria rehderiana, Scutellaria baical ensis, Oroxylum indicum, Scutellaria viscidula
Baicalein-7-O-beta-D- glucopyranoside	57396-78-8	Scutellaria baicalensis, Oroxylum indicum
Chrysin 7-O-beta-galactopyranuronoside		Centaurea pseudoscabiosa ssp. pseudoscabiosa
Baicalein-6-glucuronide	35990-03-5	Oroxylum indicum
5, 6, 4'-Trihydroxyflavone-7- O-beta-D-galactonic acid		Erigeron breviscapus
Breviscapine	116122-36-2	Erigeron breviscapus
Scutellarin	27740-01-8	Scutellaria barbata [Syn. Scutellaria rivularis], Clerodendron fragrans, Plantago major, Erigeron breviscapus, Sorbaria arborea, Scutellaria altissima, Scutellaria baicalensis, Perilla frutescens var. acuta [Syn. Perilla frutescens var. purpurascens], Oroxylum indicum, Sorbaria sorbifolia
Oroxylin A 7-O-glucuronide	36948-76-2	Scutellaria baicalensis
Baicalein	491-67-8	Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Scutellaria amoena, Scutellaria rehderiana, Scutellaria baicalensis, Scutellaria likiangensis, Oroxylum indicum, Scutellaria viscidula

Interaction between ACE2 (E) and S (A)	Interaction between atoms
E:LYS417:NZ--A:ASP30:OD2	E:NH2^s---A:O^s
A:LYS31:NZ--E:GLU484:OE1	A:NH2^s---E:O^s
E:ARG403:NH1--A:GLU37:OE2	E:NH---A:O^s
A:TYR41:OH--E:THR500:OG1	A:OH---E:O^s
A:GLN42:NE2--E:GLY446:O	A::NH2^s--E:O
A:GLN42:NE2--E:TYR449:OH	A::N^s---E:OH
A:TYR83:OH--E:ASN487:OD1	A:OH---E:O^s
E:TYR449:OH--A:ASP38:OD2	E:OH---A:O^s
E:ASN487:ND2--A:GLN24:OE1	E:NH2^s---A:O^s
E:GLY502:N--A:LYS353:O	E:NH---A:O
E:TYR505:OH--A:GLU37:OE1	E:OH---A:O
A:HIS34:CD2--E:TYR453:OH	A:C^s--E:OH
A:TYR83:OH--E:PHE486	A:OH--E:N
E:LEU455:CD1--A:HIS34	E:C--A:NH
A:LYS353:C, O; GLY354:N--E:TYR505	A:C, O; N---E:NH
E:PHE486--A:MET82	E:O--A:NH
E:TYR489--A:LYS31	E:O--A:NH
E:TYR505-A:LYS353	E:O--A:NH

Interaction between ACE2 (E) and S (A)	Interaction between atoms
E:LYS417:NZ--A:ASP30:OD2	E:NH2^s---A:O^s
A:LYS31:NZ--E:GLU484:OE1	A:NH2^s---E:O^s
E:ARG403:NH1--A:GLU37:OE2	E:NH---A:O^s
A:TYR41:OH--E:THR500:OG1	A:OH---E:O^s
A:GLN42:NE2--E:GLY446:O	A::NH2^s--E:O
A:GLN42:NE2--E:TYR449:OH	A::N^s---E:OH
A:TYR83:OH--E:ASN487:OD1	A:OH---E:O^s
E:TYR449:OH--A:ASP38:OD2	E:OH---A:O^s
E:ASN487:ND2--A:GLN24:OE1	E:NH2^s---A:O^s
E:GLY502:N--A:LYS353:O	E:NH---A:O
E:TYR505:OH--A:GLU37:OE1	E:OH---A:O
A:HIS34:CD2--E:TYR453:OH	A:C^s--E:OH
A:TYR83:OH--E:PHE486	A:OH--E:N
E:LEU455:CD1--A:HIS34	E:C--A:NH
A:LYS353:C, O; GLY354:N--E:TYR505	A:C, O; N---E:NH
E:PHE486--A:MET82	E:O--A:NH
E:TYR489--A:LYS31	E:O--A:NH
E:TYR505-A:LYS353	E:O--A:NH

Name	Activity	Reference
Camostat mesylate	Oral active protease inhibitor. It can inhibit trypsin, various inflammatory proteases, TMPRSS2, and partially block SARS-CoV-2 (COVID-19) from entering lung cells	[24]
BDBM50324475	TMPRSS2 inhibitor	[25]
BDBM50349344	TMPRSS2 inhibitor	[25]
BDBM50349345	TMPRSS2 inhibitor	[25]

Name	Activity	Reference
Camostat mesylate	Oral active protease inhibitor. It can inhibit trypsin, various inflammatory proteases, TMPRSS2, and partially block SARS-CoV-2 (COVID-19) from entering lung cells	[24]
BDBM50324475	TMPRSS2 inhibitor	[25]
BDBM50349344	TMPRSS2 inhibitor	[25]
BDBM50349345	TMPRSS2 inhibitor	[25]

基于靶点的抗新型冠状病毒病COVID-19药物发现

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刘艾林 ^* , 杜冠华 ^*

药学学报 | 聚焦抗新冠病毒药物研究 2020,55(6): 1073-1080

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药学学报 | 聚焦抗新冠病毒药物研究 2020, 55(6): 1073-1080

基于靶点的抗新型冠状病毒病COVID-19药物发现

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刘艾林^*, 杜冠华^*

作者信息

中国医学科学院、北京协和医学院药物研究所, 北京 100050

通讯作者:

*刘艾林, Tel:86-10-83150885, E-mail:liuailin@imm.ac.cn;

杜冠华, E-mail:dugh@imm.ac.cn

Drug discovery for COVID-19 treatment based on drug targets

Ai-lin LIU^*, Guan-hua DU^*

Affiliations

Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China

出版时间: 2020-06-12 doi: 10.16438/j.0513-4870.2020-0319

文章导航

摘要

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新型冠状病毒（SARS-CoV-2）引发的新冠病毒病（COVID-19），采取对症治疗不失为可行有效的治疗方案，但治疗药物大多缺乏针对性。基于病毒复制过程中的关键蛋白和病毒引发的病理机制，研制有针对性的治疗药物，将为临床提供更加有效的治疗方案。此外，由于新型冠状病毒是RNA病毒，而RNA病毒基因易于变异，因此针对新冠病毒病的新药研发将是一项长期而艰巨的任务。本文基于新型冠状病毒从吸附、进入宿主细胞到病毒复制过程中的关键蛋白及病毒感染引发的致病因素等多个环节的潜在靶点，利用分子模拟和机器学习等算法，探讨防治COVID-19新药发现的研究思路，并简述本课题组所开展的相关工作，为促进不同作用机制的新药发现提供可行性研究方法和策略。

关键词

新型冠状病毒 / 新型冠状病毒病 / 药物靶点 / 药物发现 / 分子模拟 / 机器学习

Abstract

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Coronavirus disease 2019 (COVID-19), caused by the novel coronavirus (SARS-CoV-2) is treated in accordance with symptoms, which is feasible and effective. However, current therapeutic drugs are ineffective against this virus. The development of targeted therapeutic drugs that are based on key proteins in SARS-CoV-2 replication and pathogenesis will provide a more effective means for clinical treatment. In addition, because SARS-CoV-2 is an RNA virus, which typically mutate readily, new drug development against COVID-19 will be a long-term and arduous task. New approaches to drug discovery for COVID-19 treatment using molecular simulation and machine learning algorithms, and based on the key proteins in the process of SARS-CoV-2 adsorption, entry into the host cell and viral replication are discussed herein, and we briefly introduce related work in our laboratory that can provide strategies to promote the discovery of drugs with different mechanisms of action.

Key words

novel coronavirus / novel coronavirus disease / drug target / drug discovery / molecular simulation / machine learning

引用本文

刘艾林, 杜冠华. 基于靶点的抗新型冠状病毒病COVID-19药物发现. 药学学报, 2020 , 55 (6) : 1073 -1080 . DOI: 10.16438/j.0513-4870.2020-0319

Ai-lin LIU, Guan-hua DU. Drug discovery for COVID-19 treatment based on drug targets[J]. Acta Pharmaceutica Sinica, 2020 , 55 (6) : 1073 -1080 . DOI: 10.16438/j.0513-4870.2020-0319

正文

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目前, 全球大流行的呼吸系统疾病, 其病原经分离鉴定为新型冠状病毒, 被世界卫生组织(WHO)命名为SARS-CoV-2, 由该病毒引发的疾病被命名为COVID-19^{[1, 2]}。基因测序结果表明, SARS-CoV-2与SARS-CoV和MERS-CoV一样, 同属于冠状病毒β属, 与SARS-CoV的基因序列同源性为75%~80%^[3], 与蝙蝠SARS样冠状病毒(bat-sl-coVZC45)的同源性超过85%^[4]。由于SARS-CoV-2首次在人际之间传播, 每个人对此病毒都没有免疫力, 均属易感人群。一些国家由于没有采取有效措施, 使得疫情迅速蔓延, 近乎失控。

世界各国为抗击疫情制定了各种防控措施, 其中抗新型冠状病毒病的新药发现, 是防控措施的重要组成部分, 新药发现将为临床防治新型冠状病毒病提供重要治疗方案。针对性地开展抗新型冠状病毒病的新药研发, 首先需要在了解病毒的结构组成、病毒吸附、进入宿主细胞及复制过程中的关键因素, 以及病毒感染机体后的病理特征等信息的基础上, 有针对性地开展新药发现研究, 为深入研发有发展前景的药物奠定理论基础, 提供信息依据。

1 抗新型冠状病毒的潜在药物靶点及其生物学特点

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新型冠状病毒SARS-CoV-2为单正链RNA病毒, 大约3万个碱基。病毒表面有包膜, 颗粒呈圆形、椭圆形或轻度多形性, 直径100~120 nm。冠状病毒膜表面包含刺突糖蛋白(S蛋白)、小包膜糖蛋白(E蛋白)、膜蛋白(M蛋白)^[5-7], 病毒颗粒内部还有核衣壳蛋白(N蛋白), 它们均为病毒的结构蛋白。其中, S蛋白具有受体识别和膜融合的功能, 决定病毒是否能入侵靶细胞, 是开发疫苗的重要靶点^[8]。

新型冠状病毒SARS-CoV-2同SARS-CoV和MERS-CoV一样, 均可引发人呼吸道感染, SARS-CoV主要感染纤毛支气管上皮细胞和Ⅱ型肺泡细胞, MERS-CoV主要感染未分化的支气管上皮细胞和Ⅱ型肺泡细胞。两者感染细胞类型存在差异, 与病毒受体不同有关。SARS-CoV以血管紧张素转化酶2 (angiotensin converting enzyme 2, ACE2)作为受体, 而MERS-CoV以二肽基肽酶(DPPIV)为受体。研究表明, SARS-CoV-2与SARS-CoV相同, 均以ACE2为受体感染上皮细胞^[9]。Ⅱ型肺泡细胞同时表达ACE2和跨膜丝氨酸蛋白酶(transmembrane protease serines, TMPRSS2), 其中病毒S蛋白与ACE2受体识别与吸附有关, TMPRSS2在膜融合中扮演重要角色^[10]。因此, ACE2和TMPRSS2均为潜在的抗病毒药物靶点。

病毒进入宿主细胞后, 病毒的核酸在酸化的内涵体中释放, 从而产生有效感染。病毒的正链RNA作为模板, 直接附着于宿主细胞的核糖体上翻译早期蛋白, 首先翻译出大分子多聚蛋白前体, 在细胞或病毒编码的蛋白酶作用下, 多聚蛋白被切割成功能蛋白(如RNA聚合酶)及早期结构蛋白。然后在RNA聚合酶的作用下, 以病毒正链RNA为模板, 合成互补的负链RNA, 然后以负链RNA为模板复制子代病毒RNA; 以亚基因组mRNA翻译病毒蛋白, 如衣壳蛋白、膜蛋白、非结构蛋白等, 最后进行装配。再经转运, 分泌到细胞表面, 通过胞吐作用释放子代病毒。

ORF1ab是第一个被制造出来的病毒蛋白, 它由16个蛋白(NSP1~NSP16)接合在一起。通过研究, 大部分蛋白的功能已得到初步认识, 但少数蛋白如NSP2和NSP15的功能有待于认识。其中, NSP1专门破坏宿主细胞, NSP3具有去标签和切割蛋白功能, NSP4是囊泡制造者, 用于保存病毒的RNA新拷贝, NSP5是蛋白质剪刀, 剪断大部分的NSP蛋白, NSP6是囊泡制造工厂, 与NSP3和NSP4合作制造囊泡, NSP7和NSP8辅助NSP12复制病毒, NSP10和NSP16联手将病毒基因伪装起来以逃避宿主蛋白的捕获, NSP12 (RNA polymerase)是病毒复制机, 将遗传物质组装成病毒基因组, NSP13 (RNA helicase)为解旋RNA, NSP14为病毒基因校对蛋白, NSP15具有粉碎病毒残余RNA功能^{[11, 12]}。这些非结构蛋白均具有生物学功能, 阻断其功能作用可以达到抑制病毒复制的目的, 均为潜在的药物靶点, 其中RNA聚合酶(RNA polymerase)为公认的抗病毒靶点, RNA解螺旋酶(RNA helicase)为广谱抗病毒药物的理想靶点。

此外, 病毒复制过程中, 还有很多蛋白酶参与了病毒的复制, 如3C-like蛋白酶(3-chymotrypsin-like protease, 3CLpro)负责切割病毒基因组翻译的蛋白前体, 得到多个非结构蛋白, 用于组装形成新病毒; 而木瓜样蛋白酶(papain-like protease, PLpro)是一种多功能病毒蛋白酶, 是SARS冠状病毒复制酶复合体形成的重要调节蛋白分子, 是一种病毒编码的去泛素化酶, 对细胞蛋白具有明显的去泛素化作用, 且对抗原抗病毒天然免疫反应具有负调节作用, 是冠状病毒的一种重要干扰素抑制剂; 弗林蛋白酶(Furin protease)是真核生物细胞中一种重要的内切蛋白酶, 有研究表明, SARS-CoV-2可能存在Furin蛋白酶切割位点, SARS-CoV-2在组装时, S蛋白被Furin蛋白酶切割为S1 (受体结合结构域)和S2 (膜融合结构域), 不同亚基被分泌出去, 从而侵染其他细胞^{[13, 14]}。因此, 3CLpro、PLpro和Furin均为潜在药物靶点, 可用于新药发现。

呼吸道是SARS-CoV-2感染的主要部位。当COVID-19感染呼吸道时, 可引起轻度或高度急性呼吸综合征, 随后释放促炎细胞因子, 如IL-1β和IL-6等。抑制促炎性IL-1家族成员和IL-6已被证明在许多炎症疾病, 包括病毒感染中具有治疗作用^[15]。遗体解剖报告提示, 重症患者的气道被黏液严重堵塞, 肺泡功能受损以及多器官衰竭等病理状态。实验室检查结果提示, 在发病早期, 外周血白细胞总数正常或减少, 淋巴细胞计数减少, 严重者D-二聚体升高且外周血淋巴细胞进行性减少, 提示免疫力逐渐下降。针对患者炎症损伤和免疫力下降以及黏液堵塞气道等病理特点的新药, 将为临床对症治疗提供有益的方案。病毒感染过程及其涉及的潜在靶点如表 1所示。

虽然目前临床上已有一些药物用于新冠肺炎的治疗, 但它们对于新型冠状病毒病的治疗仍缺乏针对性, 且是否有效尚无定论。因此, 有针对性地研发抗新型冠状病毒以及抗新型冠状病毒病的药物仍然迫在眉睫。本文将基于新型冠状病毒从吸附、进入宿主细胞到病毒复制过程的关键蛋白以及病毒感染引发的致病因素, 利用分子模拟、机器学习等算法, 探讨新药发现的研究策略, 并简述本实验室所开展的相关工作, 为研究发现具有不同作用机制的新药提供可行性方案。

2 靶向ACE2的药物发现

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研究发现, 与SARS-CoV相似, SARS-CoV-2也是通过ACE2进入宿主细胞的, 尤其是Ⅱ型肺泡上皮细胞。病毒的刺突蛋白S与ACE2接触后, ACE2的完整分子结构或跨膜区域与病毒一起, 通过胞吞作用进入细胞(内化)。在丝氨酸蛋白酶TMPRSS2的作用下, 病毒上的刺突蛋白进一步被激活, 发生膜融合, 病毒RNA释放入细胞质, 完成病毒对细胞的感染^{[1, 16-19]}, 因此, ACE2是预防冠状病毒感染的重要靶点, 也是预防新型冠状病毒病进一步恶化的重要靶点。

在应急用药的情况下, 通过基因序列比对, 发现同源性较高的冠状疾毒, 从而借鉴已有病毒病的临床用药和用药方案。通过与SARS-CoV和MERS-CoV基因序列比对, 发现SARS-CoV-2与SARS-CoV的同源性比MERS-CoV的更高。通过S蛋白的结构建模, 发现人类细胞上ACE2受体与SARS-CoV-2的S蛋白有更强的相互作用^[20], 这一预测结果得到两个冷冻电镜技术应用成果的验证^{[21, 22]}。

据报道, 黄芩苷、橙皮苷和甘草酸苷等天然化合物是ACE2潜在抑制剂, 可用于预防SARS-CoV感染^[23]。由于SARS-CoV-2与SARS-CoV的基因序列同源性高, 且ACE2是它们共同的受体, 因此推测黄芩苷、橙皮苷和甘草酸苷等天然化合物对SARS-CoV-2的感染也有潜在的预防作用。根据相似性的结构具有相似性活性的原则, 本课题组利用分子相似性分析方法, 从天然产物数据库中预测发现了230个与黄芩苷、橙皮苷和甘草酸苷结构相似度大于0.75的天然化合物。表 2列出相似度较高的前25个天然化合物及其植物来源等信息, 由于参考文献太多, 这里不作赘述。这些天然化合物抑制ACE2的活性, 还有待于后续实验评价。表中天然化合物的中药或植物来源信息, 将为中药的临床用药提供有益的信息依据。

3 靶向病毒刺突糖蛋白S与ACE2受体蛋白相互作用的药物发现

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S蛋白, 全称spike glycoprotein (刺突糖蛋白), 位于新冠病毒最外层, 是一种三聚体Ⅰ类融合蛋白。作为新型冠状病毒附着并感染人类的核心蛋白, S蛋白是开发疫苗、治疗性抗体和药物的关键靶点。

目前, S蛋白结构以及S蛋白与ACE2受体蛋白相互作用复合物结构已有报道^[20-23]。Wrapp等^[20]利用冷冻电镜(Cryo-EM), 成功解析了新型冠状病毒S蛋白结构。Yan等^[21]利用冷冻电镜测定了全长人ACE2与中性氨基酸转运体B0AT1的复合物结构, 报道了在B0AT1存在下, SARS-CoV-2的RBD结合全长人ACE2的冷冻电镜结构, 分析了RBD与受体之间的相互作用。Lan等^[23]利用X射线衍射技术, 解析了新型冠状病毒表面刺突糖蛋白受体结合区与人受体ACE2蛋白复合物的晶体结构, 并准确定位出新型冠状病毒RBD和受体ACE2的相互作用位点, 阐明了新型冠状病毒刺突糖蛋白介导细胞侵染的结构基础及分子机制, 为疫苗设计、治疗性抗体药物开发以及小分子药物发现奠定了坚实的基础。研究发现, 新型冠状病毒的RBD与ACE2胞外域结合的亲和力要比SARS冠状病毒高出10~20倍。新型冠状病毒与ACE2之间的高亲和力, 推动其在人际之间快速传播, 这也为新冠病毒的超强传染性提供了合理的解释。

基于X射线衍射技术得到的病毒S蛋白-人受体ACE2蛋白复合物晶体结构, 利用分子模拟软件分析蛋白表面的相互作用, 可以观察到S蛋白受体结合区与ACE2之间的非键相互作用的氨基酸残基, 详细信息见表 3。从表中可以看出, S蛋白与ACE2蛋白之间的识别来自于氨基酸残基侧链原子之间的非键相互作用, 以及两个蛋白主链原子之间的非键相互作用。如果化合物能够与ACE2(E)上这些关键氨基酸相互作用, 就可以有效阻止S蛋白与ACE2的相互作用, 从而阻止病毒的有效识别和感染。

为了寻找发现抑制S-ACE2结合的有效药物, 可将S-ACE2的结合部位设为活性腔, 通过分子对接计算和对接后分析, 发现与ACE2亲和力较强的化合物, 在此基础上分析化合物是否与ACE2(E)上的这些关键氨基酸相互作用, 相互作用的氨基酸越多, 视为抑制S-ACE2结合的潜力越大, 越有利于阻止病毒的侵入。

4 靶向TMPRSS2蛋白酶的药物发现

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最新研究发现, SARS-CoV-2病毒侵入细胞不仅依赖于ACE2受体, 还需要细胞丝氨酸蛋白酶TMPRSS2, 并发现临床使用的TMPRSS2蛋白酶抑制剂可以阻断SARS-CoV-2病毒侵入细胞, 为新型冠状病毒病的治疗带来新思路^[24]。

实验结果表明, 丝氨酸蛋白酶TMPRSS2抑制剂甲磺酸卡莫司他(camostat mesylate), 能够部分阻断SARS-CoV-2的S蛋白驱动的对Caco-2和Vero-TMPRSS2细胞的感染。Camostat mesylate可以显著降低SARS-CoV-2对人肺细胞的感染。

研究表明, 冠状病毒刺突蛋白S促进了病毒对宿主细胞的吸附, 且在病毒膜蛋白与细胞膜融合过程中, TMPRSS2促进S蛋白打开入侵细胞的大门, 对于病毒的有效感染发挥了重要作用, 丝氨酸蛋白酶的活性对于病毒的传播和宿主的感染是必要条件。

总之, SARS-CoV-2需要利用TMPRSS2蛋白酶的活性, 使病毒包膜与细胞膜融合, 而TMPRSS2抑制剂camostat mesylate可以阻断SARS-CoV-2对肺细胞的感染, 提示TMPRSS2是抗新型冠状病毒病的重要靶点, 可用于抗新型冠状病毒病药物的发现, 为防治新型冠状病毒病提供新的治疗方案。

有关TMPRSS2抑制剂的研究报道较少, 表 4^{[24, 25]}中列举了4个抑制剂的信息, 可为TMPRSS2抑制剂设计、药效团的构建与新药发现提供信息参考。

5 靶向病毒复制酶及复制相关蛋白酶的药物发现

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RNA依赖的RNA聚合酶(RNA dependent polymerase, RdRp)主要负责病毒RNA的复制, 是研究广谱抗病毒药物靶点。病毒复制相关的蛋白酶如PLpro、3CLpro、弗林蛋白酶(Furin protease)等蛋白酶在病毒蛋白的加工和多聚蛋白的裂解中发挥重要作用, 因此均可作为潜在药物靶点, 用于抗病毒的药物发现^[26-28]。

病毒复制及复制相关蛋白酶抑制剂属于广谱抗病毒药物。目前临床上用于治疗新型冠状病毒病的药物中, 瑞德西韦和利巴韦林均为RNA聚合酶抑制剂, 而匹洛那韦和利托那韦均为3CLpro抑制剂^[29]。体外细胞水平的抗新型冠状病毒实验结果提示, 在10 μg·mL^-1检测浓度为下, 瑞德西韦和利巴韦林使病毒载量分别降低3个数量级和1个数量级, 说明它们均可有效抑制病毒的复制, 瑞德西韦的活性比利巴韦林强2个数量级。洛匹那韦/利托那韦(克力芝)临床具有一定疗效, 也证明3CLpro抑制剂对新冠病毒病有一定的治疗作用。然而, 由于RNA病毒易于变异的特性, 新药发现的任务仍然紧迫而艰巨。

经RCSB PDB、BindingDB等数据库检索和文献调研, 已收集了冠状病毒相关的不同蛋白酶的晶体结构, 可用于分子对接计算。RdRp、PLpro、3CLpro和Furin蛋白酶的抑制剂数量分别为48、71、174和461个。

对于靶点结构已知、配体分子较多的这些靶点, 可基于配体结构, 利用机器学习算法, 构建化合物-靶点相互作用预测模型, 模型验证之后, 可用于化合物数据库的大规模虚拟筛选, 以发现新的潜在活性化合物, 工作流程见图 1^{[30, 31]}。目前本课题组已针对病毒RdRp、PLpro、3Clpro和Furin等靶点抑制剂, 建立了一系列的化合物-靶点相互作用预测模型, 已用于化合物数据库的虚拟筛选, 预测得到的潜在活性化合物, 正在进行细胞水平的抗新型冠状病毒实验验证。

6 靶向宿主的药物发现

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病毒侵入宿主细胞后, 在细胞内大量繁殖, 从而产生大量炎症因子和氧自由基, 损伤靶器官, 如引起肺部呼吸衰竭, 出现过度免疫并产生过多炎症因子。而对于重症患者的某些检测指标, 如淋巴细胞进行性减少, 说明免疫力渐进性下降。在没有特效药的情况下, 依据患者的临床症状进行对症治疗是临床治疗的重要策略。

中药在对症治疗的临床应用中表现尤为突出, 本文不作详细阐述。中药有效成分的组合“甘草酸二铵+维生素C+芦丁”的鸡尾酒疗法也具有其合理性, 其中甘草酸二铵具有抗炎症瀑布的作用, 但没有皮质激素的不良反应, 维生素C作为抗氧化剂, 结合甘草酸的抗炎作用, 对于组织器官将发挥双重保护作用。而芦丁具有抗炎、维持血管抵抗力、降低通透性等作用, 可有效减少炎性物质向组织器官的渗透^[32]。

病毒感染引发的炎症因子有很多, 主要包括IL-6、IL-1β和TNF-α等^[33]。本课题组针对病毒感染引发的炎症因子, 利用机器学习算法, 已系统地建立了化合物-靶点相互作用预测模型, 并已用于活性化合物的预测^[34]。预测模型的应用, 一方面可以发现新的抗炎药物, 另一方面也适用于一些抗炎药物的作用机制的研究。

干扰素在机体的抗病毒免疫中具有重要作用, 在抗新型冠状病毒的临床治疗方案中已经纳入α-干扰素, 其疗效是肯定的。本课题组与中国疾控中心病毒病预防控制所通力合作, 对致力于防治病毒性疾病药物开发的北京某知名企业提供的不同类型干扰素, 进行了抗新型冠状病毒SARS-CoV-2的体外活性评价, 发现其中一些干扰素具有明显的体外抗病毒活性, 在10 μg·mL^-1的细胞实验浓度下, 病毒载量可降低数百倍, 提示其具有很好的应用前景。

7 结语与展望

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基于病毒复制过程各个环节的关键蛋白及宿主来源的关键蛋白, 研发单靶点药物, 将为联合用药提供新的治疗方案, 而同时靶向多个病毒靶点和宿主靶点的药物则是理想的多靶点抗病毒药物, 也是抗病毒新药研发的热点。中药方剂多成分、多靶点、多通路的物质基础和网络机制具有天然的治疗优势, 将为中医药的发展注入新的活力^[35]。基于不同药物靶点和基于不同药物来源的新药研发将为未来防治COVID-19提供更多可行的有针对性的有效治疗方案。

自新型冠状病毒病暴发以来, 世界各国科学家、医学工作者、政府机构、企事业单位等各部门各行业人员, 通力合作, 对于新型冠状病毒的基因测序、临床检测、靶点研究、药物开发、疫苗研制、疾病治疗等各方面均取得阶段性成果^[36-38]。目前对于SARS-CoV-2及COVID-19虽然已有一定的认识, 但仍然有很多未知。随着各项工作的不断推进, 特别是对SARS-CoV-2病毒蛋白结构及功能的不断阐明, 以及对COVID-19疾病认识的不断深入, 必将有力地促进抗COVID-19的新药发现, 为COVID-19提供更多有效的治疗方案。

基金

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“十三五”重大新药创制专项(2018ZX09711001-003-002)
“十三五”重大新药创制专项(2018ZX09711001-012)
国家自然科学基金资助项目(81673480)
协和创新工程项目(2016-I2M-3-007)

参考文献

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文献

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2020年第55卷第6期

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文章信息

doi: 10.16438/j.0513-4870.2020-0319

接收时间：2020-03-13
首发时间：2026-01-21
出版时间：2020-06-12

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文章信息

作者

出版历史

收稿日期：2020-03-13
修回日期：2020-04-17

基金

“十三五”重大新药创制专项(2018ZX09711001-003-002)

“十三五”重大新药创制专项(2018ZX09711001-012)

国家自然科学基金资助项目(81673480)

协和创新工程项目(2016-I2M-3-007)

作者信息

中国医学科学院、北京协和医学院药物研究所, 北京 100050

通讯作者:

*刘艾林, Tel:86-10-83150885, E-mail:liuailin@imm.ac.cn;

杜冠华, E-mail:dugh@imm.ac.cn

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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TxT

Process of virus infection	Potential target	Potential effect of drug
Virus adsorption	ACE2, S-ACE2	Interference the recognition for ACE2 receptor and suppress virus invasion, therefore they can be used for COVID prevention
Membrane fusion	TMPRSS2	Inhibit the fusion between membrane protein of virus and cell membrane, protect cells against SARS-CoV-2 induced COVID-19
Non-structural protein	NSP1, NSP3, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12 (RNA dependent RNA polymerase, RdRp), NSP13 (RNA helicase), NSP15, NSP16	Inhibit the activity of non-structural functional proteins and interference the procedure of virus replication. Used for COVID-19 to inhibit virus replication
Virus replication	PLpro, 3CLpro, Furin	Inhibit the activity of different enzymes in virus replication process
Pathogenic factors	IL-6, IL-1β, TNF-α	Anti-inflammation, immunity regulation

Process of
virus infection

Potential target

Potential effect of drug

Virus adsorption

ACE2, S-ACE2

Interference the recognition for ACE2 receptor and suppress virus
invasion, therefore they can be used for COVID prevention

Membrane fusion

TMPRSS2

Inhibit the fusion between membrane protein of virus and cell
membrane, protect cells against SARS-CoV-2 induced COVID-19

Non-structural
protein

NSP1, NSP3, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12 (RNA dependent RNA polymerase, RdRp), NSP13 (RNA helicase), NSP15, NSP16

Inhibit the activity of non-structural functional proteins and
interference the procedure of virus replication. Used for COVID-19 to inhibit virus replication

Virus replication

PLpro, 3CLpro, Furin

Inhibit the activity of different enzymes in virus replication process

Pathogenic factors

IL-6, IL-1β, TNF-α

Anti-inflammation, immunity regulation

English name	CAS	Source
Hesperidin	520-26-3	Galium aparine, Citrus medica var. sarcodactylis, Citrus chachiensis, Poncirus trifoliata, Capsella bursa-pastoris, Citrus tankan, Schizonepeta tenuifolia [Syn. Nepeta tenuifolia], Citrus reticulata, Citrus medica, Citrus limonia, Microtoena prainiana, Citrus limon, Mentha longifolia, Galium mollugo, Citrus unshiu, Citrus aurantium
Isosakuranetin-7-rutinoside		Citrus sinensis
Narirutin	14259-46-2	Citrus sinensis, Citrus sp.
Diosmin	520-27-4	Galium aparine, Sophora viciifolia, Toddalia asiatica [Syn. Toddalia aculeata; Paullinia asiatica], Citrus medica var. sarcodactylis, Scabiosa caucasica, Capsella bursa-pastoris, Scrophularia nodosa, Rosmarinus officinalis, Citrus limon, Urtica cannabina, Zanthoxylum nitidum, Zanthoxylum avicennae
Neohesperidin	13241-33-3	Poncirus trifoliata, Citrus limon, Citrus unshiu, Citrus aurantium
Chrysoeriol 7-O-rutinoside		Saussurea medusa
Acaciin	480-36-4	Chrysanthemum boreale, Robinia pseudoacacia, Trapa bispinosa, Agastache rugosus, Buddleja officinalis, Chrysanthemum indicum
Ombuoside	20188-85-6	Phytolacca dioica, Gynostemma pentaphyllum, Flyriella parryi
Uralsaponin A	103000-77-7	Glycyrrhiza uralensis, Glycyrrhiza kansuensis
Licoricesaponin H2	135815-61-1	Glycyrrhiza uralensis, Glycyrrhiza inflata
Glycyrrhizic acid	1405-86-3	Glycyrrhiza aspera, Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Abrus precatorius, Glycyrrhiza inflata
Licoricesaponin G2	118441-84-2	Glycyrrhiza uralensis, Glycyrrhiza inflata
Glyeurysaponin	137476-70-1	Glycyrrhiza kansuensis
Araboglycyrrhizin	121687-83-0	Glycyrrhiza inflata
Uralsaponin B	105038-43-5	Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Glycyrrhiza inflata
Licoricesaponin J2	134250-13-8	Glycyrrhiza uralensis
Baicalin	21967-41-9	Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Salvia miltiorrhiza, Scutellaria amoena, Scutellaria rehderiana, Scutellaria baical ensis, Oroxylum indicum, Scutellaria viscidula
Baicalein-7-O-beta-D- glucopyranoside	57396-78-8	Scutellaria baicalensis, Oroxylum indicum
Chrysin 7-O-beta-galactopyranuronoside		Centaurea pseudoscabiosa ssp. pseudoscabiosa
Baicalein-6-glucuronide	35990-03-5	Oroxylum indicum
5, 6, 4'-Trihydroxyflavone-7- O-beta-D-galactonic acid		Erigeron breviscapus
Breviscapine	116122-36-2	Erigeron breviscapus
Scutellarin	27740-01-8	Scutellaria barbata [Syn. Scutellaria rivularis], Clerodendron fragrans, Plantago major, Erigeron breviscapus, Sorbaria arborea, Scutellaria altissima, Scutellaria baicalensis, Perilla frutescens var. acuta [Syn. Perilla frutescens var. purpurascens], Oroxylum indicum, Sorbaria sorbifolia
Oroxylin A 7-O-glucuronide	36948-76-2	Scutellaria baicalensis
Baicalein	491-67-8	Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Scutellaria amoena, Scutellaria rehderiana, Scutellaria baicalensis, Scutellaria likiangensis, Oroxylum indicum, Scutellaria viscidula

English name

CAS

Source

Hesperidin

520-26-3

Galium aparine, Citrus medica var. sarcodactylis, Citrus chachiensis, Poncirus trifoliata, Capsella
bursa-pastoris, Citrus tankan, Schizonepeta tenuifolia [Syn. Nepeta tenuifolia], Citrus reticulata,
Citrus medica, Citrus limonia, Microtoena prainiana, Citrus limon, Mentha longifolia, Galium mollugo, Citrus unshiu, Citrus aurantium

Isosakuranetin-7-rutinoside

Citrus sinensis

Narirutin

14259-46-2

Citrus sinensis, Citrus sp.

Diosmin

520-27-4

Galium aparine, Sophora viciifolia, Toddalia asiatica [Syn. Toddalia aculeata; Paullinia asiatica],
Citrus medica var. sarcodactylis, Scabiosa caucasica, Capsella bursa-pastoris, Scrophularia nodosa, Rosmarinus officinalis, Citrus limon, Urtica cannabina, Zanthoxylum nitidum, Zanthoxylum avicennae

Neohesperidin

13241-33-3

Poncirus trifoliata, Citrus limon, Citrus unshiu, Citrus aurantium

Chrysoeriol 7-O-rutinoside

Saussurea medusa

Acaciin

480-36-4

Chrysanthemum boreale, Robinia pseudoacacia, Trapa bispinosa, Agastache rugosus, Buddleja
officinalis, Chrysanthemum indicum

Ombuoside

20188-85-6

Phytolacca dioica, Gynostemma pentaphyllum, Flyriella parryi

Uralsaponin A

103000-77-7

Glycyrrhiza uralensis, Glycyrrhiza kansuensis

Licoricesaponin H2

135815-61-1

Glycyrrhiza uralensis, Glycyrrhiza inflata

Glycyrrhizic acid

1405-86-3

Glycyrrhiza aspera, Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Abrus
precatorius, Glycyrrhiza inflata

Licoricesaponin G2

118441-84-2

Glycyrrhiza uralensis, Glycyrrhiza inflata

Glyeurysaponin

137476-70-1

Glycyrrhiza kansuensis

Araboglycyrrhizin

121687-83-0

Glycyrrhiza inflata

Uralsaponin B

105038-43-5

Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza kansuensis, Glycyrrhiza inflata

Licoricesaponin J2

134250-13-8

Glycyrrhiza uralensis

Baicalin

21967-41-9

Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Salvia miltiorrhiza, Scutellaria
amoena, Scutellaria rehderiana, Scutellaria baical ensis, Oroxylum indicum, Scutellaria viscidula

Baicalein-7-O-beta-D-
glucopyranoside

57396-78-8

Scutellaria baicalensis, Oroxylum indicum

Chrysin 7-O-beta-galactopyranuronoside

Centaurea pseudoscabiosa ssp. pseudoscabiosa

Baicalein-6-glucuronide

35990-03-5

Oroxylum indicum

5, 6, 4'-Trihydroxyflavone-7-
O-beta-D-galactonic acid

Erigeron breviscapus

Breviscapine

116122-36-2

Erigeron breviscapus

Scutellarin

27740-01-8

Scutellaria barbata [Syn. Scutellaria rivularis], Clerodendron fragrans, Plantago major, Erigeron
breviscapus, Sorbaria arborea, Scutellaria altissima, Scutellaria baicalensis, Perilla frutescens var.
acuta [Syn. Perilla frutescens var. purpurascens], Oroxylum indicum, Sorbaria sorbifolia

Oroxylin A 7-O-glucuronide

36948-76-2

Scutellaria baicalensis

Baicalein

491-67-8

Scutellaria scordifolia, Scutellaria hypericifolia, Plantago major, Scutellaria amoena, Scutellaria
rehderiana, Scutellaria baicalensis, Scutellaria likiangensis, Oroxylum indicum, Scutellaria viscidula

Interaction between ACE2 (E) and S (A)	Interaction between atoms
E:LYS417:NZ--A:ASP30:OD2	E:NH2^s---A:O^s
A:LYS31:NZ--E:GLU484:OE1	A:NH2^s---E:O^s
E:ARG403:NH1--A:GLU37:OE2	E:NH---A:O^s
A:TYR41:OH--E:THR500:OG1	A:OH---E:O^s
A:GLN42:NE2--E:GLY446:O	A::NH2^s--E:O
A:GLN42:NE2--E:TYR449:OH	A::N^s---E:OH
A:TYR83:OH--E:ASN487:OD1	A:OH---E:O^s
E:TYR449:OH--A:ASP38:OD2	E:OH---A:O^s
E:ASN487:ND2--A:GLN24:OE1	E:NH2^s---A:O^s
E:GLY502:N--A:LYS353:O	E:NH---A:O
E:TYR505:OH--A:GLU37:OE1	E:OH---A:O
A:HIS34:CD2--E:TYR453:OH	A:C^s--E:OH
A:TYR83:OH--E:PHE486	A:OH--E:N
E:LEU455:CD1--A:HIS34	E:C--A:NH
A:LYS353:C, O; GLY354:N--E:TYR505	A:C, O; N---E:NH
E:PHE486--A:MET82	E:O--A:NH
E:TYR489--A:LYS31	E:O--A:NH
E:TYR505-A:LYS353	E:O--A:NH

Interaction between
ACE2 (E) and S (A)

Interaction between
atoms

E:LYS417:NZ--A:ASP30:OD2

E:NH2^s---A:O^s

A:LYS31:NZ--E:GLU484:OE1

A:NH2^s---E:O^s

E:ARG403:NH1--A:GLU37:OE2

E:NH---A:O^s

A:TYR41:OH--E:THR500:OG1

A:OH---E:O^s

A:GLN42:NE2--E:GLY446:O

A::NH2^s--E:O

A:GLN42:NE2--E:TYR449:OH

A::N^s---E:OH

A:TYR83:OH--E:ASN487:OD1

A:OH---E:O^s

E:TYR449:OH--A:ASP38:OD2

E:OH---A:O^s

E:ASN487:ND2--A:GLN24:OE1

E:NH2^s---A:O^s

E:GLY502:N--A:LYS353:O

E:NH---A:O

E:TYR505:OH--A:GLU37:OE1

E:OH---A:O

A:HIS34:CD2--E:TYR453:OH

A:C^s--E:OH

A:TYR83:OH--E:PHE486

A:OH--E:N

E:LEU455:CD1--A:HIS34

E:C--A:NH

A:LYS353:C, O; GLY354:N--E:TYR505

A:C, O; N---E:NH

E:PHE486--A:MET82

E:O--A:NH

E:TYR489--A:LYS31

E:O--A:NH

E:TYR505-A:LYS353

E:O--A:NH

Name	Activity	Reference
Camostat mesylate	Oral active protease inhibitor. It can inhibit trypsin, various inflammatory proteases, TMPRSS2, and partially block SARS-CoV-2 (COVID-19) from entering lung cells	[24]
BDBM50324475	TMPRSS2 inhibitor	[25]
BDBM50349344	TMPRSS2 inhibitor	[25]
BDBM50349345	TMPRSS2 inhibitor	[25]

Name

Structure

Activity

Reference

Camostat mesylate

Oral active protease inhibitor. It can inhibit trypsin, various
inflammatory proteases, TMPRSS2, and partially block SARS-CoV-2 (COVID-19) from entering lung cells

[24]

BDBM50324475

TMPRSS2 inhibitor

[25]

BDBM50349344

TMPRSS2 inhibitor

[25]

BDBM50349345

TMPRSS2 inhibitor

[25]