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WO2021203701A1 - Application of sofosbuvir in preparation of drug for preventing and treating coronavirus - Google Patents

Application of sofosbuvir in preparation of drug for preventing and treating coronavirus Download PDF

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WO2021203701A1
WO2021203701A1 PCT/CN2020/129175 CN2020129175W WO2021203701A1 WO 2021203701 A1 WO2021203701 A1 WO 2021203701A1 CN 2020129175 W CN2020129175 W CN 2020129175W WO 2021203701 A1 WO2021203701 A1 WO 2021203701A1
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coronavirus
cov
sofosbuvir
preparation
sars
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Chinese (zh)
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袁曙光
邹荣峰
罗木鹏
陈显翀
崔文强
赵金存
孙静
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中国科学院深圳先进技术研究院
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A61K31/7072Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00Drugs for disorders of the respiratory system
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12Antivirals
    • A61P31/14Antivirals for RNA viruses

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  • the African green monkey kidney cells (VeroE6) were inoculated into 96-well plates at 3 ⁇ 10 5 cells/well, and placed in the minimum Eagle's medium (MEM; GibcoInvitrogen) containing 10% fetal bovine serum (FBS; GibcoInvitrogen). Incubate at 37°C with 5% CO 2 until the monolayer grows up. A 100-fold dilution of the novel coronavirus clinical isolates of the novel coronavirus pneumonia was inoculated into a 96-well plate full of monolayer cells, and cultured at 37°C and 5% CO 2 for two days (including the normal control group).
  • MEM minimum Eagle's medium
  • FBS fetal bovine serum

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  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
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  • General Chemical & Material Sciences (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
  • Pulmonology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

An application of sofosbuvir in preparation of a drug for preventing and treating coronavirus. Particularly, disclosed is an application of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomer, ester, amide or prodrug thereof in preparation of a drug for preventing and/or treating diseases caused by coronavirus. The coronaviruses are novel coronaviruses SARS-Cov-2, SARS-CoV, HCoV 229E, NL63, OC43, HKU1 and MERS-CoV. The median effective concentration of sofosbuvir on novel coronavirus of Coronavirus Disease 2019 is 1.12 μM, the toxicity is low, and a good treatment window is provided.

Description

索非布韦在制备预防和治疗冠状病毒的药物中的应用Application of Sofosbuvir in the Preparation of Drugs for Prevention and Treatment of Coronavirus 技术领域Technical field
本发明属于抗病毒药物领域,具体涉及索非布韦在制备预防和治疗抗冠状病毒的药物中的应用。The invention belongs to the field of antiviral drugs, and specifically relates to the application of sofosbuvir in the preparation of drugs for the prevention and treatment of anti-coronaviruses.
背景技术Background technique
新冠肺炎(COVID-19)是由新型冠状病毒(SARS-Cov-2)感染人体而引起的传染性疾病,其症状主要包括发热、乏力、干咳、呼吸困难和肾衰竭等[The Lancet,2020,395(10223):507-513;The Lancet,2020,395(10223):497-506]。新冠肺炎于2019年12月开始有报道病例,随后全球多个国家,包括韩国、日本、泰国、伊朗、新加坡、德国、法国和美国等也陆续出现病例。冠状病毒在系统分类上属冠状病毒科(Coronaviridae)冠状病毒属(Corona virus)。冠状病毒属的病毒是具外套膜(envelope)的正链单股RNA病毒,直径约80~120nm,其遗传物质是所有RNA病毒中最大的,一般只会感染人、鼠、猪、猫、犬、禽类脊椎动物。冠状病毒于1937年被首次从鸡身上分离出来。冠状病毒粒子形状并不规则,直径约60~220nm。病毒具有包膜结构,上面有三种蛋白:刺突糖蛋白(S,Spike Protein)、小包膜糖蛋白(E,Envelope Protein)和膜糖蛋白(M,Membrane Protein),少数种类还有血凝素糖蛋白(HE蛋白,Haemaglutinin-esterase)[Nederlands Tijdschrift Voor Geneeskunde,2014,158(158):A8119-A8119]。New coronary pneumonia (COVID-19) is an infectious disease caused by a new type of coronavirus (SARS-Cov-2) infecting the human body. Its symptoms mainly include fever, fatigue, dry cough, dyspnea, and kidney failure. [The Lancet, 2020, 395(10223):507-513; The Lancet, 2020,395(10223):497-506]. New crown pneumonia began to report cases in December 2019, and then many countries around the world, including South Korea, Japan, Thailand, Iran, Singapore, Germany, France, and the United States, have also seen cases one after another. Coronaviruses belong to the genus Coronavirus in the family Coronaviridae (Coronaviridae). The virus of the genus Coronavirus is a positive-stranded single-stranded RNA virus with an envelope, with a diameter of about 80-120nm. Its genetic material is the largest among all RNA viruses, and generally only infects humans, mice, pigs, cats, and dogs. , Poultry vertebrates. The coronavirus was first isolated from chickens in 1937. Coronavirus particles are irregular in shape, about 60-220nm in diameter. The virus has an envelope structure with three proteins: Spike Protein (S, Spike Protein), Small Envelope Protein (E, Envelope Protein) and Membrane Protein (M, Membrane Protein), and a few types also have hemagglutination. Glycoprotein (HE protein, Haemaglutinin-esterase) [Nederlands Tijdschrift Voor Geneeskunde, 2014, 158(158): A8119-A8119].
SARS-Cov-2病毒颗粒直径在60~140nm之间,包膜外有9~12nm的尖刺,形似花冠。基因组测序表明,SARS-Cov-2是一种单链RNA冠状病毒。通过与其他病毒样品基因序列的比较,发现SARS-Cov-2与SARS-CoV(79.5%)[Nature,2020] 和蝙蝠冠状病毒(96%)相似[bioRxiv,2020,2020.01.22.914952],并推测该病毒可能起源于蝙蝠[bioRxiv,2020,2020.01.24.915157;Nature,2020]。SARS-Cov-2病毒属于βCoV,是区别于SARS-CoV和MERS-CoV的HCoV家族的第7个成员[New England Journal of Medicine,2020],其余6个成员包括HCoV 229E、NL63、OC43、HKU1、SARS-CoV和MERS-CoV。The diameter of SARS-Cov-2 virus particles is between 60nm and 140nm, and there are 9-12nm spikes outside the envelope, resembling a corolla. Genome sequencing showed that SARS-Cov-2 is a single-stranded RNA coronavirus. By comparison with the gene sequences of other virus samples, it is found that SARS-Cov-2 is similar to SARS-CoV (79.5%) [Nature,2020] and bat coronavirus (96%) [bioRxiv,2020,2020.01.22.914952], and speculates The virus may originate from bats [bioRxiv, 2020, 2020.01.24.915157; Nature, 2020]. SARS-Cov-2 virus belongs to βCoV, which is the seventh member of the HCoV family that is different from SARS-CoV and MERS-CoV [New England Journal of Medicine, 2020]. The remaining 6 members include HCoV 229E, NL63, OC43, and HKU1 , SARS-CoV and MERS-CoV.
引起新型冠状病毒肺炎的是一种新型冠状病毒,它与人们熟知的引起非典型性肺炎的SARS-CoV同属冠状病毒,但类型不同,其致死率虽低于SARS-CoV但传染性远远高于SARS-CoV。全球传播表明新型冠状病毒肺炎疫情仍存在扩散的可能。开发有效的抗病毒药物和疫苗成为当下最为紧迫的任务。目前关于COVID-19的治疗主要依靠对症辅助治疗,尚缺乏有效的特效药物和疫苗。然而,新药开发和疫苗的制备是一项耗时的过程,不仅研发周期长,上市批准后的批量生产亦是一项耗时的工程。从上市药物中寻找具有抗SARS-Cov-2病毒感染作用的“老药”,是治疗和预防COVID-19这种爆发性感染疾病的一种有效策略。目前正在研究的抗SARS-Cov-2病毒药物主要包括RNA聚合酶抑制剂、β干扰素、单克隆抗体以及疫苗等。然而,到目前为止,尚无确切有效的抗病毒药物和疫苗。从对这些药物的体外抗病毒活性数据的分析来看,大多数药物的抗病毒IC 50处于低微摩尔至中微摩尔之间,体内疗效尚在观察之中。 The new type of coronavirus pneumonia is caused by a new type of coronavirus. It is the same type of coronavirus as SARS-CoV, which is known to cause atypical pneumonia, but is of a different type. Although its fatality rate is lower than SARS-CoV, it is far more infectious. In SARS-CoV. The global spread shows that the new coronavirus pneumonia epidemic still has the possibility of spreading. The development of effective antiviral drugs and vaccines has become the most urgent task at the moment. At present, the treatment of COVID-19 mainly relies on symptomatic adjuvant treatment, and there is still a lack of effective specific drugs and vaccines. However, the development of new drugs and the preparation of vaccines is a time-consuming process. Not only is the research and development cycle long, mass production after market approval is also a time-consuming project. Searching for "old drugs" with anti-SARS-Cov-2 virus infection effects from marketed drugs is an effective strategy for the treatment and prevention of COVID-19, an explosive infectious disease. The anti-SARS-Cov-2 virus drugs currently under study mainly include RNA polymerase inhibitors, beta interferon, monoclonal antibodies, and vaccines. However, so far, there are no definite and effective antiviral drugs and vaccines. Data from the analysis of antiviral activity of these drugs in vitro, the majority of IC 50 antiviral drugs is in between the low micromolar to micromolar, still observed in vivo efficacy.
索非布韦由吉利德(Gilead)研发,于2013年12月6日获美国食品药品管理局(FDA)批准上市,之后于2014年1月16日获得欧洲药物管理局(EMA)批准上市,后又于2015年3月26日获得日本医药品医疗器械综合机构(PMDA)批准上市,由吉利德上市销售,商品名为
Figure PCTCN2020129175-appb-000001
索非布韦是一种HCV NS5B RNA依赖型RNA聚合酶抑制剂,该聚合酶对病毒复制有重要作用。鉴于该药物的作用机 制,并且在药物虚拟筛选结果的基础上我们开展了索非布韦在新冠肺炎新型冠状病毒抗病毒药效的应用性研究。
Sofosbuvir was developed by Gilead and was approved for marketing by the U.S. Food and Drug Administration (FDA) on December 6, 2013, and then approved by the European Medicines Agency (EMA) on January 16, 2014. Later, it was approved for listing by the Japan Pharmaceuticals and Medical Devices Agency (PMDA) on March 26, 2015, and was marketed by Gilead under the trade name
Figure PCTCN2020129175-appb-000001
Sofosbuvir is an inhibitor of HCV NS5B RNA-dependent RNA polymerase, which plays an important role in virus replication. In view of the drug's mechanism of action, and on the basis of the virtual screening results of the drug, we have carried out an applied research on the antiviral efficacy of sofosbuvir in the new coronavirus of new coronary pneumonia.
开发有效的冠状病毒肺炎治疗特效药物成为了当下一个迫在眉睫需要解决的课题。在针对新型冠状病毒(SARS-Cov-2)病毒药物研发方面,本发明研究了索非布韦抗SARS-Cov-2病毒作用。The development of effective drugs for the treatment of coronavirus pneumonia has become an urgent issue that needs to be resolved. In the research and development of new coronavirus (SARS-Cov-2) virus drugs, the present invention studies the anti-SARS-Cov-2 virus effect of Sofosbuvir.
发明内容Summary of the invention
本发明的目的在于提供索非布韦在制备预防或治疗冠状病毒所致疾病的药物中的应用。The purpose of the present invention is to provide the application of sofosbuvir in the preparation of drugs for preventing or treating diseases caused by coronavirus.
具体而言,为解决本发明的技术问题,采用如下技术方案:Specifically, in order to solve the technical problems of the present invention, the following technical solutions are adopted:
本发明提供了索非布韦或其可药用的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药在制备预防和/或治疗冠状病毒所致疾病的药物中的应用。The present invention provides sofosbuvir or its pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs in the preparation of prevention and/or treatment of coronary Application in medicines for diseases caused by viruses.
在本发明的技术方案中,所述的冠状病毒为新型冠状病毒SARS-Cov-2、SARS-CoV、HCoV 229E、NL63、OC43、HKU1和MERS-CoV。In the technical solution of the present invention, the coronaviruses are the new coronaviruses SARS-Cov-2, SARS-CoV, HCoV 229E, NL63, OC43, HKU1 and MERS-CoV.
在本发明的技术方案中,冠状病毒所致疾病为SARS-Cov-2、SARS-CoV、HCoV 229E、NL63、HCoV-OC43、HKU1或MERS-CoV任一病毒引起的感染性疾病或其并发症;优选为呼吸道感染疾病,例如,严重急性呼吸综合征、严重急性呼吸道综合征冠状病毒2型、中东呼吸综合症。In the technical solution of the present invention, the disease caused by the coronavirus is an infectious disease or its complications caused by any virus of SARS-Cov-2, SARS-CoV, HCoV 229E, NL63, HCoV-OC43, HKU1 or MERS-CoV ; Preferably it is a respiratory infection disease, for example, severe acute respiratory syndrome, severe acute respiratory syndrome coronavirus type 2, and Middle East respiratory syndrome.
在本发明的技术方案中,索非布韦如结构式(1)所示的In the technical scheme of the present invention, sofosbuvir is shown in structural formula (1)
Figure PCTCN2020129175-appb-000002
Figure PCTCN2020129175-appb-000002
在本发明的技术方案中,索非布韦或其可药用的盐、同位素、立体异构体、 立体异构体的混合物、互变异构体、酯、酰胺或前药作为唯一活性成分在制备预防和/或治疗冠状病毒所致疾病的药物中的应用。In the technical solution of the present invention, sofosbuvir or its pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs are used as the sole active ingredient Application in the preparation of medicines for preventing and/or treating diseases caused by coronaviruses.
在本发明的技术方案中,索非布韦或其可药用的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药,与其他抗病毒药物制备的组合物作为活性成分在制备预防和/或治疗冠状病毒所致疾病的药物中的应用。In the technical solution of the present invention, sofosbuvir or its pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs, and other anti- The application of the composition prepared by the virus medicine as an active ingredient in the preparation of a medicine for preventing and/or treating diseases caused by coronavirus.
在本发明的技术方案中,其他抗病毒药物选自其他抗病毒药物选自更昔洛韦、阿昔洛韦、金刚烷胺、金刚乙胺、奥司他韦、阿巴卡韦、醋孟南、阿昔洛韦钠、阿德福韦、阿洛夫定、阿韦舒托、盐酸三环癸胺、阿拉诺丁、阿立酮、阿替韦啶甲磺酸酯、阿夫立定、西多福韦、西潘茶碱、恩曲他滨、盐酸阿糖胞苷、甲磺酸地拉韦啶、地昔洛韦、去羟肌苷、二噁沙利、依度尿苷、乙米韦林、依曲西他平、恩韦拉登、恩韦肟、贺普丁、泛昔洛韦、盐酸氯苯氢异喹、非西他滨、非阿尿苷、磷利酯、膦乙酸钠、甘西洛维钠、碘苷、茚地那韦、乙氧丁酮醛、拉米夫定、洛布卡韦、洛德腺苷、洛匹那韦、盐酸美莫汀、甲红硫脲、那非那韦、奈韦拉平、喷昔洛韦、吡罗达韦、利巴韦林、甲磺酸沙奎那韦、利托那韦、盐酸索金刚胺、索立夫定、匍枝青霉菌素、司他夫定、替诺福韦、盐酸梯络龙、曲氟尿苷、盐酸伐昔洛韦、阿糖腺苷、磷酸阿糖腺苷、阿糖腺苷磷酸钠、替拉那韦、韦罗肟、扎西他滨、齐多夫定、净韦肟。In the technical scheme of the present invention, other antiviral drugs are selected from other antiviral drugs selected from ganciclovir, acyclovir, amantadine, rimantadine, oseltamivir, abacavir, acetomeng South, Acyclovir Sodium, Adefovir, Alovudine, Avirsuto, Tricyclodecylamine Hydrochloride, Aranodine, Aridone, Ateviridine Mesylate, Afridine, Cidofovir, Cipanphylline, Emtricitabine, Cytarabine Hydrochloride, Delavirdine Mesylate, Deciclovir, Didanosine, Dioxali, Edouridine, Beta Mivirin, Etracitabine, Enviraden, Enviroxime, Hepdin, Famciclovir, Chlorphenhydrazine Hydrochloride, Fecitabine, Fe-Auridine, Phospritide, Phosphonoacetate Sodium, Glycine Silovir sodium, iodoside, indinavir, ethoxybutone aldehyde, lamivudine, lobukavir, lordadenosine, lopinavir, memotine hydrochloride, methyl red thiourea, that Finavir, nevirapine, penciclovir, pirodavir, ribavirin, saquinavir mesylate, ritonavir, somantine hydrochloride, solivudine, penicillin, sclerotium Tavudine, Tenofovir, Tirolone Hydrochloride, Trifluridine, Valaciclovir Hydrochloride, Vidarabine, Vidarabine Phosphate, Vidarabine Sodium Phosphate, Tiranavir, Vero Oxime, zalcitabine, zidovudine, virgin oxime.
本发明另一个方面提供了一种治疗或预防冠状病毒科病毒所致疾病的药物组合物,所述药物组合物中包含索非布韦或其可药用的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药。Another aspect of the present invention provides a pharmaceutical composition for the treatment or prevention of diseases caused by coronaviruses of the coronavirus family, the pharmaceutical composition comprising sofosbuvir or its pharmaceutically acceptable salts, isotopes, stereoisomers, Mixtures of stereoisomers, tautomers, esters, amides or prodrugs.
在本发明的技术方案中,药物组合物的还包括药学上可接受的辅料。In the technical scheme of the present invention, the pharmaceutical composition also includes pharmaceutically acceptable excipients.
在本发明的技术方案中,药物组合物的剂型为口服制剂、肺部吸入制剂、粘 膜给药制剂、眼用制剂或注射剂。In the technical scheme of the present invention, the dosage form of the pharmaceutical composition is oral preparation, pulmonary inhalation preparation, mucosal administration preparation, ophthalmic preparation or injection.
在本发明的技术方案中,口服制剂选自颗粒剂、粉末剂、丸剂、片剂、胶囊或口服液。In the technical scheme of the present invention, the oral preparation is selected from granules, powders, pills, tablets, capsules or oral liquids.
本发明另一个方面提供了索非布韦作为抗冠状病毒科病毒的消毒剂的用途。Another aspect of the present invention provides the use of sofosbuvir as a disinfectant against coronaviruses of the coronavirus family.
本发明另一个方面提供了一种消除冠状病毒科病毒污染的消毒剂,所述消毒剂包括索非布韦。Another aspect of the present invention provides a disinfectant for eliminating coronavirus pollution, the disinfectant includes sofosbuvir.
本发明另一个方面提供了一种用于治疗冠状病毒科病毒所致疾病的方法,包括将治疗有效量的索非布韦或其药物学上可接受的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药给药于受试者。Another aspect of the present invention provides a method for the treatment of diseases caused by coronaviruses, comprising adding a therapeutically effective amount of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, stereo A mixture of isomers, tautomers, esters, amides or prodrugs is administered to the subject.
本发明另一个方面提供了一种用于预防受试者感染冠状病毒科病毒的方法,包括将治疗有效量的索非布韦或其药物学上可接受的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯或前药给药在感染前给予受试者。Another aspect of the present invention provides a method for preventing a subject from being infected with a coronavirus of the coronavirus family, which comprises adding a therapeutically effective amount of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, Administration of mixtures of stereoisomers, tautomers, esters or prodrugs is administered to the subject prior to infection.
在本发明的技术方案中,所述药物或药物组合物中不包含糖皮质激素或盐皮质激素。In the technical solution of the present invention, the drug or pharmaceutical composition does not contain glucocorticoid or mineralocorticoid.
有益效果Beneficial effect
本发明首次发现索非布韦对新型冠状病毒有抗病毒作用,可阻断新型冠状病毒感染宿主细胞,可用作治疗抗冠肺病毒感染方面的疾病治疗。The present invention finds for the first time that sofosbuvir has an antiviral effect on the novel coronavirus, can block the infection of host cells by the novel coronavirus, and can be used as a disease treatment for the treatment of anti-coronavirus infection.
索非布韦对新冠肺炎新型冠状病毒(SARS-Cov-2)的半数有效浓度(EC 50)为1.12μM,半数有效浓度低,毒性低,具有一个良好的治疗窗口。 Sofosbuvir has a median effective concentration (EC 50 ) of 1.12 μM for SARS-Cov-2, a low half effective concentration, low toxicity, and a good therapeutic window.
具体实施方式Detailed ways
为了使本发明的上述目的、特征和优点能够更加明显易懂,下面结合附图对本发明的具体实施方式做详细的说明,但不能理解为对本发明的可实施范围的限 定。In order to make the above objectives, features and advantages of the present invention more obvious and understandable, the specific embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings, but they should not be understood as limiting the scope of implementation of the present invention.
实施例1病毒扩增Example 1 Virus amplification
将非洲绿猴肾细胞(VeroE6)按3×10 5个/孔,接种至96孔板中,在含有10%胎牛血清(FBS;GibcoInvitrogen)的minimum Eagle’smedium(MEM;GibcoInvitrogen)中,置37℃,5%CO 2培养,待长满单层。将新冠肺炎新型冠状病毒临床分离株100倍稀释接种到长满单层细胞的96孔板中,置37℃,5%CO 2培养两天(含正常对照组)。 The African green monkey kidney cells (VeroE6) were inoculated into 96-well plates at 3×10 5 cells/well, and placed in the minimum Eagle's medium (MEM; GibcoInvitrogen) containing 10% fetal bovine serum (FBS; GibcoInvitrogen). Incubate at 37°C with 5% CO 2 until the monolayer grows up. A 100-fold dilution of the novel coronavirus clinical isolates of the novel coronavirus pneumonia was inoculated into a 96-well plate full of monolayer cells, and cultured at 37°C and 5% CO 2 for two days (including the normal control group).
两天后病变程度高达75%以上,置于-80℃超低温冰箱,反复冻融一次,收集细胞扩增的病毒液,3000r/min离心30分钟,去沉淀物,分装小管置-80℃超低温冰箱长期保存。Two days later, the degree of disease was as high as 75%, placed in a -80℃ ultra-low temperature refrigerator, repeated freezing and thawing once, collecting the amplified virus solution, centrifuged at 3000r/min for 30 minutes, removing the sediment, and placing the small tubes in the -80℃ ultra-low temperature refrigerator Long-term preservation.
实施例2索非布韦药物毒性评价Example 2 Evaluation of Sofosbuvir Drug Toxicity
索非布韦粉末用DMSO溶解后,加入培养液稀释至20mg/mL,DMSO终浓度为1%,经0.22μm滤膜过滤后置4℃保存;过滤后置4℃保存。按每孔约2.5×10 4细胞接种到96孔板,24~48h后待细胞长成单层后,弃去培养液,加入不同稀释度的药物100μL/孔,正常细胞对照孔加入100μL/孔MEM,37℃ 5%CO 2继续培养2~5天,每孔加CCK8法溶液(5mg/mL)20μL,置37℃ 5%CO 2保温箱中继续孵育4小时。吸弃培养上清液,每孔加100μL二甲基亚砜(DMSO),低速振荡10分钟,使结晶物充分融解。选择490nm波长,在酶联免疫监测仪上测定各孔光吸收值。通过毒性评价可知药物添加浓度达到400μM时,依然未见明显细胞毒性,说明药物的细胞毒性低,具有良好的治疗窗口。 After sofossbuvir powder is dissolved in DMSO, add culture broth to dilute to 20mg/mL, the final concentration of DMSO is 1%, filter through 0.22μm membrane and store at 4℃; after filtration, store at 4℃. Inoculate about 2.5×10 4 cells per well into a 96-well plate. After 24 to 48 hours, when the cells grow into a monolayer, discard the culture medium, add 100μL/well of drugs of different dilutions, and 100μL/well for normal cell control wells. MEM, 37°C and 5% CO 2 continue to incubate for 2 to 5 days, add 20 μL of CCK8 solution (5 mg/mL) to each well, and place in a 37°C 5% CO 2 incubator for further incubation for 4 hours. Aspirate and discard the culture supernatant, add 100 μL of dimethyl sulfoxide (DMSO) to each well, and shake at low speed for 10 minutes to fully melt the crystals. Choose the wavelength of 490nm, and measure the light absorption value of each hole on the enzyme-linked immunosorbent monitor. Through the toxicity evaluation, it can be seen that when the concentration of the drug added reaches 400 μM, there is still no obvious cytotoxicity, indicating that the cytotoxicity of the drug is low and it has a good therapeutic window.
实施例3索非布韦抗新冠肺炎新型冠状病毒药效评价Example 3 Evaluation of Sofosbuvir's Anti-New Coronary Pneumonia New Coronavirus Drug Effect
为了评估药物的抗病毒效力,将VeroE6细胞在密度为5×10 4细胞/孔的48孔 细胞培养皿中培养过夜。添加病毒(MOI为0.05)使其感染2小时。然后加入2倍梯度稀释的药物,每个浓度设置4个复孔,以200μM为药物起始浓度,在34℃、5%CO 2培养箱中孵育2天。记录细胞病变(Cytopathogenic Effect,CPE。细胞出现CPE按6级标准记录。记录CPE后,用CCK8法染色,进行OD值测定。用Reed—Muench法计算药物半数有效浓度(EC 50)为1.12μM。通过实验结果可知膦甲酸钠抗病毒的半数有效浓度低,抗病毒效果好。 In order to evaluate the antiviral efficacy of the drug, VeroE6 cells were cultured overnight in a 48-well cell culture dish with a density of 5×10 4 cells/well. Virus (MOI of 0.05) was added to make it infected for 2 hours. Then add 2 times the gradient dilution of the drug, each concentration is set with 4 replicate wells, with 200 μM as the initial concentration of the drug, incubate for 2 days in a 34°C, 5% CO 2 incubator. Cytopathogenic Effect (CPE) was recorded. The occurrence of CPE in cells was recorded according to the 6-level standard. After recording CPE, it was stained with the CCK8 method and the OD value was determined. The half effective concentration (EC 50 ) of the drug was calculated by the Reed-Muench method to be 1.12 μM. The experimental results show that foscarnet sodium has a low antiviral half effective concentration and good antiviral effect.
实施例4免疫荧光检测Example 4 Immunofluorescence detection
免疫荧光显微镜:为了检测VeroE6细胞中病毒蛋白的表达,将细胞用4%多聚甲醛固定并用0.5%TritonX-100透化。然后在室温下用5%牛血清白蛋白(BSA)封闭细胞2小时。将细胞与一抗(蝙蝠SARS相关冠状病毒的病毒核衣壳蛋白多克隆抗体,anti-NP,以1:1000稀释度)一起孵育2小时,然后与二抗孵育抗体(488AffiniPure Donkey Anti-Rabbit IgG(H+L)。细胞核用Hoechst33258染料(Beyotime,China)染色。通过荧光显微镜观察可知索非布韦能有效杀灭细胞中的病毒,并且对细胞影响较小,具有良好的治疗窗口。Immunofluorescence microscopy: In order to detect the expression of viral proteins in VeroE6 cells, the cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% TritonX-100. The cells were then blocked with 5% bovine serum albumin (BSA) for 2 hours at room temperature. Incubate the cells with the primary antibody (polyclonal antibody against the nucleocapsid protein of the bat SARS-related coronavirus, anti-NP, at a dilution of 1:1000) for 2 hours, and then incubate the antibody with the secondary antibody (488AffiniPure Donkey Anti-Rabbit IgG) (H+L). The nucleus is stained with Hoechst33258 dye (Beyotime, China). Observation by fluorescence microscope shows that Sofosbuvir can effectively kill the virus in the cell, and has little effect on the cell, and has a good treatment window.

Claims (10)

  1. 索非布韦或其可药用的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药在制备预防和/或治疗冠状病毒所致疾病的药物中的应用。Sofosbuvir or its pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs are used in the preparation of prevention and/or treatment of diseases caused by coronavirus The application of drugs.
  2. 根据权利要求1所述的用途,所述的冠状病毒为新型冠状病毒SARS-Cov-2、SARS-CoV、HCoV 229E、NL63、OC43、HKU1和MERS-CoV。The use according to claim 1, wherein the coronaviruses are new coronaviruses SARS-Cov-2, SARS-CoV, HCoV 229E, NL63, OC43, HKU1 and MERS-CoV.
  3. 根据权利要求1所述的用途,冠状病毒所致疾病为SARS-Cov-2、SARS-CoV、HCoV 229E、NL63、HCoV-OC43、HKU1或MERS-CoV任一病毒引起的感染性疾病或其并发症;感染性疾病优选为呼吸道感染疾病。The use according to claim 1, the disease caused by the coronavirus is an infectious disease caused by any virus of SARS-Cov-2, SARS-CoV, HCoV 229E, NL63, HCoV-OC43, HKU1, or MERS-CoV or its complications The infectious disease is preferably a respiratory tract infection disease.
  4. 根据权利要求1所述的用途,索非布韦或其可药用的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药作为唯一活性成分在制备预防和/或治疗冠状病毒所致疾病的药物中的应用;或者The use according to claim 1, sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture of stereoisomers, tautomers, esters, amides or prodrugs thereof as the sole active The application of the ingredients in the preparation of drugs for the prevention and/or treatment of diseases caused by coronavirus; or
    索非布韦或其可药用的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药,与其他抗病毒药物的组合物作为活性成分在制备预防和/或治疗冠状病毒所致疾病的药物中的应用;Sofosbuvir or its pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs, in combination with other antiviral drugs as active ingredients Application in the preparation of medicines for the prevention and/or treatment of diseases caused by coronavirus;
    优选地,其他抗病毒药物选自更昔洛韦、阿昔洛韦、金刚烷胺、金刚乙胺、奥司他韦、阿巴卡韦、醋孟南、阿昔洛韦钠、阿德福韦、阿洛夫定、阿韦舒托、盐酸三环癸胺、阿拉诺丁、阿立酮、阿替韦啶甲磺酸酯、阿夫立定、西多福韦、西潘茶碱、恩曲他滨、盐酸阿糖胞苷、甲磺酸地拉韦啶、地昔洛韦、去羟肌苷、二噁沙利、依度尿苷、乙米韦林、依曲西他平、恩韦拉登、恩韦肟、贺普丁、泛昔洛韦、盐酸氯苯氢异喹、非西他滨、非阿尿苷、磷利酯、膦甲酸钠、膦乙酸钠、甘西洛维钠、碘苷、茚地那韦、乙氧丁酮醛、拉米夫定、洛布卡韦、洛德腺苷、洛匹那韦、盐酸美莫汀、甲红硫脲、那非那韦、奈韦拉平、喷昔洛韦、吡罗达韦、 利巴韦林、甲磺酸沙奎那韦、利托那韦、盐酸索金刚胺、索立夫定、匍枝青霉菌素、司他夫定、替诺福韦、盐酸梯络龙、曲氟尿苷、盐酸伐昔洛韦、阿糖腺苷、磷酸阿糖腺苷、阿糖腺苷磷酸钠、替拉那韦、韦罗肟、扎西他滨、齐多夫定、净韦肟。Preferably, the other antiviral drugs are selected from ganciclovir, acyclovir, amantadine, rimantadine, oseltamivir, abacavir, acemonam, acyclovir sodium, adefo Vir, alovudine, avirsuto, tricyclodecylamine hydrochloride, alanordine, aridone, ateviridine mesylate, afridine, cidofovir, sipanphylline, en Tritabine, Cytarabine Hydrochloride, Delaviridine Mesylate, Deciclovir, Didanosine, Dioxali, Edouridine, Emivirin, Etracitapine, Enemy Veraden, Enviroxime, Hepudin, Famciclovir, Chlorophenhydroisoquine Hydrochloride, Fecitabine, Feiridine, Phosphatide, Sodium Foscarnet, Phosphonoacetate Sodium, Ganciclovir Sodium, Iodoside, Indinavir, ethoxybutanone, lamivudine, lobukavir, lordadenosine, lopinavir, memotine hydrochloride, methyl red thiourea, narfinavir, nevirapine, penxix Lovir, Pirotavir, Ribavirin, Saquinavir Mesylate, Ritonavir, Somantine Hydrochloride, Sorivudine, Penicillin, Stavudine, Tenofovir , Tirolone Hydrochloride, Trifluridine, Valacyclovir Hydrochloride, Vidarabine, Vidarabine Phosphate, Vidarabine Sodium Phosphate, Tiranavir, Verooxime, Zalcitabine, Qi Dovudine, Jingwei oxime.
  5. 一种治疗或预防冠状病毒所致疾病的药物组合物,所述药物组合物中包含索非布韦或其可药用的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药。A pharmaceutical composition for treating or preventing diseases caused by coronavirus, said pharmaceutical composition comprising sofosbuvir or its pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, and interconversions Isomer, ester, amide or prodrug.
  6. 根据权利要求5所述的药物组合物,药物组合物的还包括药学上可接受的辅料。The pharmaceutical composition according to claim 5, further comprising pharmaceutically acceptable excipients.
  7. 根据权利要求5所述的药物组合物,药物组合物的剂型为口服制剂、肺部吸入制剂、粘膜给药制剂、眼用制剂或注射剂;优选地,口服制剂选自颗粒剂、粉末剂、丸剂、片剂、胶囊或口服液。The pharmaceutical composition according to claim 5, wherein the dosage form of the pharmaceutical composition is an oral preparation, a pulmonary inhalation preparation, a mucosal administration preparation, an ophthalmic preparation or an injection; preferably, the oral preparation is selected from granules, powders, and pills , Tablets, capsules or oral liquid.
  8. 一种消除冠状病毒科病毒污染的消毒剂,所述消毒剂包括索非布韦。A disinfectant for eliminating the pollution of coronaviruses of the coronavirus family, and the disinfectant includes sofosbuvir.
  9. 一种用于治疗或预防冠状病毒科病毒所致疾病的方法,包括将治疗有效量的索非布韦或其药物学上可接受的盐、同位素、立体异构体、立体异构体的混合物、互变异构体、酯、酰胺或前药给药于受试者。A method for treating or preventing diseases caused by coronaviruses of the coronavirus family, comprising adding a therapeutically effective amount of sofosbuvir or a pharmaceutically acceptable salt, isotope, stereoisomer, or mixture of stereoisomers , Tautomers, esters, amides or prodrugs are administered to the subject.
  10. 根据权利要求5-7任一项所述的药物组合物或权利要求8所述的消毒剂或权利要求9所述的方法,其中冠状病毒为新型冠状病毒SARS-Cov-2、SARS-CoV、HCoV 229E、NL63、OC43、HKU1和MERS-CoV。The pharmaceutical composition according to any one of claims 5-7 or the disinfectant according to claim 8 or the method according to claim 9, wherein the coronavirus is a new coronavirus SARS-Cov-2, SARS-CoV, HCoV 229E, NL63, OC43, HKU1 and MERS-CoV.
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Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021155119A2 (en) * 2020-01-29 2021-08-05 The Trustees Of Columbia University In The City Of New York Therapeutics for covid-19
CN111467363A (en) * 2020-04-07 2020-07-31 中国科学院深圳先进技术研究院 Application of sofosbuvir in preparation of medicine for preventing and treating coronavirus
CN112237584B (en) * 2020-08-21 2021-10-01 中国医学科学院医药生物技术研究所 Use of compounds for the prophylaxis and/or treatment of diseases caused by coronavirus infection
WO2022053164A1 (en) * 2020-09-14 2022-03-17 Hc Pharma Ag Antiviral combination for the treatment of covid-19 infection
WO2022088037A1 (en) * 2020-10-30 2022-05-05 中国科学院深圳先进技术研究院 Application of sirtinol in preparation of drug for preventing and treating coronavirus
WO2022088038A1 (en) * 2020-10-30 2022-05-05 中国科学院深圳先进技术研究院 Application of cay10603 in preparation of drugs for preventing and treating coronavirus-related diseases
CN112661801A (en) * 2020-12-18 2021-04-16 成都阿奇生物医药科技有限公司 Nucleoside analogue and deuteron thereof, and preparation method and application thereof

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101918425A (en) * 2007-03-30 2010-12-15 法莫赛特股份有限公司 Nucleoside phosphoramidate prodrugs
CN104327137A (en) * 2014-11-07 2015-02-04 王彩琴 Deuterated Sofosbuvir and application thereof
CN105705511A (en) * 2013-04-12 2016-06-22 艾其林医药公司 Deuterated nucleoside prodrugs useful for treating HCV
CN109843900A (en) * 2016-08-19 2019-06-04 桑多斯股份公司 Sofosbuvir derivatives for the treatment of hepatitis c
CN109862884A (en) * 2016-08-12 2019-06-07 桑多斯股份公司 Solid composite medicament comprising amorphous Suo Feibuwei
CN111467363A (en) * 2020-04-07 2020-07-31 中国科学院深圳先进技术研究院 Application of sofosbuvir in preparation of medicine for preventing and treating coronavirus

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101918425A (en) * 2007-03-30 2010-12-15 法莫赛特股份有限公司 Nucleoside phosphoramidate prodrugs
CN105705511A (en) * 2013-04-12 2016-06-22 艾其林医药公司 Deuterated nucleoside prodrugs useful for treating HCV
CN104327137A (en) * 2014-11-07 2015-02-04 王彩琴 Deuterated Sofosbuvir and application thereof
CN109862884A (en) * 2016-08-12 2019-06-07 桑多斯股份公司 Solid composite medicament comprising amorphous Suo Feibuwei
CN109843900A (en) * 2016-08-19 2019-06-04 桑多斯股份公司 Sofosbuvir derivatives for the treatment of hepatitis c
CN111467363A (en) * 2020-04-07 2020-07-31 中国科学院深圳先进技术研究院 Application of sofosbuvir in preparation of medicine for preventing and treating coronavirus

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHIEN MINCHEN, ANDERSON THOMAS K., JOCKUSCH STEFFEN, TAO CHUANJUAN, KUMAR SHIV, LI XIAOXU, RUSSO JAMES J., KIRCHDOERFER ROBERT N.,: "Nucleotide Analogues as Inhibitors of SARS-CoV-2 Polymerase", BIORXIV, 20 March 2020 (2020-03-20), XP055841078, Retrieved from the Internet <URL:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7239050/pdf/nihpp-2020.03.18.997585.pdf> [retrieved on 20210914], DOI: 10.1101/2020.03.18.997585 *
ELFIKY ABDO A.: "Anti-HCV, Nucleotide Inhibitors, Repurposing against COVID-19", LIFE SCIENCES, vol. 248, 28 February 2020 (2020-02-28), XP086095279, DOI: 10.1016/j.lfs.2020.117477 *
MIN JUNG SUN, KIM GEON-WOO, KWON SUNOH, JIN YOUNG-HEE: "A Cell-Based Reporter Assay for Screening Inhibitors of MERS Coronavirus RNA-Dependent RNA Polymerase Activity", JOURNAL OF CLINICAL MEDICINE, vol. 9, no. 8, pages 2399, XP055857169, DOI: 10.3390/jcm9082399 *

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