WO2021174741A1 - 二甲基小檗胺类化合物在抑制SARS-CoV-2中的用途 - Google Patents

二甲基小檗胺类化合物在抑制SARS-CoV-2中的用途 Download PDF

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WO2021174741A1
WO2021174741A1 PCT/CN2020/101581 CN2020101581W WO2021174741A1 WO 2021174741 A1 WO2021174741 A1 WO 2021174741A1 CN 2020101581 W CN2020101581 W CN 2020101581W WO 2021174741 A1 WO2021174741 A1 WO 2021174741A1
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pharmaceutically acceptable
general formula
compound represented
hydrate
solvate
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PCT/CN2020/101581
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English (en)
French (fr)
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钟武
秦成峰
周辛波
邓永强
胡季强
金庆平
余斌
曹瑞源
王曼丽
李晓峰
张娜娜
樊士勇
胡志红
李松
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中国人民解放军军事科学院军事医学研究院
浙江金华康恩贝生物制药有限公司
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Priority to JP2022552854A priority Critical patent/JP2023516706A/ja
Priority to EP20923195.0A priority patent/EP4124337A4/en
Priority to US17/908,617 priority patent/US20230119764A1/en
Publication of WO2021174741A1 publication Critical patent/WO2021174741A1/zh

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    • 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
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4748—Quinolines; Isoquinolines forming part of bridged ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives

Definitions

  • This application belongs to the field of chemical medicine, and specifically relates to dimethylberberamine compounds represented by the following formula I, their geometric isomers or their pharmaceutically acceptable salts and/or both solvates and/or both hydrates , And a pharmaceutical composition containing the above-mentioned compound for the treatment of SARS-CoV-2 infection.
  • the dimethyl berbamine compound represented by formula I is derived from the tetrandrane family plant tetrandrine ( Stephania tetrandra) bibenzyl isoquinoline alkaloids.
  • the compound of formula (I) is also called Tetrandrine, also known as tetrandrine and tetrandrine. It is an antihypertensive drug. It is mainly used clinically to treat mild and moderate hypertension, and can also be used for hypertensive crisis. . In addition, it can also be used for rheumatism, arthralgia, and neuralgia. Combined with low-dose radiation for lung cancer; also used for simple silicosis stage I, II, III and various stages of coal silicosis.
  • Coronavirus is a single-stranded, unsegmented, positive-stranded RNA virus that has an envelope and has a wide range of animal hosts.
  • SARS coronavirus and MERS coronavirus which are derived from animal infectious diseases, can cause human deaths, and they broke out in 2002 and 2012 respectively (Zaki et al., 2012; Zhong et al., 2003; Cui et al., 2019) .
  • the 2019 Novel Coronavirus (2019-nCoV) is a new strain of coronavirus that has never been found in humans before.
  • COVID-19 the official classification of the 2019 novel coronavirus (2019-nCoV) is called severe acute respiratory syndrome coronavirus 2, SARS-CoV- 2).
  • SARS-CoV- 2 severe acute respiratory syndrome coronavirus 2
  • WHO World Health Organization
  • the clinical manifestations of COVID-19 patients are that more than 90% of patients have fever, 80% of patients have dry cough, 20% of patients have shortness of breath, 15% of patients have dyspnea, and the most important manifestation is With the reduction of white blood cells and lymphocytes, the lungs of patients have varying degrees of organ damage, which has the potential risk of developing pulmonary fibrosis.
  • the virus is highly contagious.
  • tetrandrine can effectively inhibit SARS-CoV-2 virus, prevent or treat pulmonary fibrosis, thereby providing effective prevention and/or treatment of lung diseases or symptoms caused by SARS-CoV-2 virus.
  • New ways and choices. This application is now completed based on the above research.
  • This application relates to the compound represented by the general formula I for the prevention and/or treatment of SARS-CoV-2 related lung diseases or symptoms or asymptomatic or symptomatic SARS-CoV-2 infection, and its geometric isomers Or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate, or containing the compound represented by general formula I, its geometric isomer or its pharmaceutically acceptable salt and/or its solvate And/or a pharmaceutical composition of its hydrate,
  • This application also relates to the prevention and/or treatment of lung diseases associated with SARS-CoV-2 and associated with hypertension, or symptoms or asymptomatic or symptomatic SARS-CoV-2 infections associated with hypertension.
  • the compound represented by formula I, its geometric isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate, or the compound represented by formula I, its geometric isomer or its hydrate Pharmaceutical compositions of pharmaceutically acceptable salts and/or solvates and/or hydrates thereof,
  • This application also relates to the prevention and/or treatment of COVID-19 or COVID-19 with high blood pressure, or as a SARS-CoV-2 inhibitor, or for inhibiting SARS-CoV-2 in cells (such as mammalian cells).
  • the compound represented by general formula I replicated or reproduced in ), its geometric isomers or pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, or containing the compound represented by general formula I, Pharmaceutical compositions of geometric isomers or pharmaceutically acceptable salts and/or solvates and/or hydrates thereof,
  • This application further relates to the compound represented by general formula I, its geometric isomers or pharmaceutically acceptable salts and/or solvates thereof and/or its solvates and/or Hydrate, or a pharmaceutical composition containing the compound represented by the general formula I, its geometric isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate.
  • the present invention further relates to a compound represented by general formula I, its geometric isomers or a pharmaceutically acceptable salt thereof, and/or its geometrical isomers or pharmaceutically acceptable salts thereof for preventing and/or treating diseases or infections caused by coronavirus and accompanied by hypertension Solvates and/or hydrates thereof, or pharmaceutical compositions containing compounds represented by formula I, geometric isomers thereof, or pharmaceutically acceptable salts thereof, and/or solvates and/or hydrates thereof.
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate, or the compound represented by the general formula I, its geometric isomer
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is used to inhibit the replication or propagation of coronavirus in cells (for example, mammalian cells).
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is used as a coronavirus inhibitor.
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is used in the preparation of products for the prevention and/or treatment of diseases or infections caused by coronaviruses.
  • the present invention further relates to a compound represented by general formula I, its geometric isomers or pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, or a compound represented by general formula I, its geometrical isomers
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is used for the prevention and/or treatment of diseases or infections caused by coronavirus and accompanied by high blood pressure. Use in the product.
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is prepared for the prevention and/or treatment of lung diseases associated with coronavirus and accompanied by hypertension or Symptoms of product use.
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is used in the preparation of a product for inhibiting the replication or reproduction of coronavirus in cells (e.g., mammalian cells) use.
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric Use of the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate in the preparation of a product as a coronavirus inhibitor.
  • This application further relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salt and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is being prepared for the prevention and/or treatment of lung diseases or symptoms related to SARS-CoV-2, Or use in products with asymptomatic or symptomatic SARS-CoV-2 infection.
  • This application also relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salts and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric isomers
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is used to prevent and/or treat the lungs associated with SARS-CoV-2 and accompanied by hypertension. It can be used in products that cause diseases or symptoms, or asymptomatic or symptomatic SARS-CoV-2 infection with high blood pressure.
  • This application also relates to the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salts and/or its solvates and/or its hydrates, or the compound represented by the general formula I, its geometric isomers
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is used in the preparation of products for the prevention and/or treatment of COVID-19 or COVID-19 accompanied by high blood pressure Or in the preparation of products as SARS-CoV-2 inhibitors, or in the preparation of products for inhibiting SARS-CoV-2 from replicating or multiplying in cells (such as mammalian cells).
  • This application further relates to a method for preventing and/or treating a disease or infection caused by a coronavirus or a disease or infection caused by a coronavirus accompanied with high blood pressure, which includes administering to a host in need a preventive and/or effective amount of
  • This application further relates to methods for preventing and/or treating lung diseases or symptoms related to coronavirus or lung diseases or symptoms related to coronavirus and accompanied by high blood pressure, which include administering to a host in need of preventing and/ Or an effective amount of the compound represented by general formula I, its geometric isomers or its pharmaceutically acceptable salts and/or its solvates and/or its hydrates, or containing the compound represented by general formula I, its geometrical isomers
  • the pharmaceutical composition of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate are examples of the isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate.
  • This application further relates to a method for inhibiting the replication or reproduction of coronavirus in a host in need, which includes administering to the host in need a preventive and/or effective amount of a compound represented by general formula I, its geometric isomers or its A pharmaceutically acceptable salt and/or a solvate and/or a hydrate thereof, or a compound represented by formula I, a geometric isomer thereof, or a pharmaceutically acceptable salt and/or a solvate thereof Or the pharmaceutical composition of its hydrate.
  • This application also relates to a method for preventing and/or treating SARS-CoV-2 related pulmonary diseases or symptoms or asymptomatic or symptomatic SARS-CoV-2 infection, which includes a preventive and/or effective amount of general formula
  • the compound represented by I, its geometrical isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate, or the compound represented by the general formula I, its geometrical isomer or its pharmacy The pharmaceutical composition of the above-acceptable salt and/or its solvate and/or its hydrate is administered to patients with SARS-CoV-2 related lung diseases or symptoms or suffering from asymptomatic or symptomatic SARS-CoV- 2 Infected host.
  • This application also relates to methods for preventing and/or treating SARS-CoV-2 related lung diseases or symptoms with hypertension or asymptomatic or symptomatic SARS-CoV-2 infection with hypertension, which includes Preventive and/or effective amounts of the compound represented by general formula I, its geometric isomers or pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, or containing compounds represented by general formula I
  • the pharmaceutical composition of the compound, its geometrical isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate is administered to a lung disease or symptom related to SARS-CoV-2 and suffering from hypertension Or an asymptomatic or symptomatic SARS-CoV-2 infected host with hypertension.
  • This application also relates to a method for treating and/or preventing COVID-19 or COVID-19 with hypertension in a host in need, or a method for inhibiting SARS-CoV-2 replication or reproduction in a host in need, the method
  • This includes administering to a host in need a therapeutically and/or prophylactically effective amount of the compound represented by formula I, its geometric isomers or pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, or A pharmaceutical composition containing the compound represented by the general formula I, its geometric isomers or its pharmaceutically acceptable salts and/or its solvates and/or its hydrates.
  • the coronavirus described in this application is SARS-CoV-2.
  • the coronavirus described in this application is SARS-CoV-2.
  • the disease caused by the coronavirus described in this application is COVID-19.
  • the disease associated with high blood pressure caused by the coronavirus described in the present application is COVID-19 with high blood pressure.
  • the lung disease or symptom related to the coronavirus described in the present application is a lung disease or symptom related to SARS-CoV-2.
  • the pulmonary disease or symptom associated with the coronavirus and associated with hypertension described in this application is a lung disease or symptom associated with SARS-CoV-2 and associated with hypertension.
  • the infection caused by the coronavirus described in this application is asymptomatic or symptomatic SARS-CoV-2 infection.
  • the infection caused by the coronavirus described in the present application accompanied by hypertension is asymptomatic or symptomatic SARS-CoV-2 infection accompanied by hypertension.
  • the product described in this application is a human drug or a veterinary drug for animals.
  • the product described in this application is a solid preparation, an injection, a spray, a liquid preparation, an inhalation preparation, or a compound preparation.
  • the host described in this application is a mammal.
  • the mammals described in this application include bovines, equines, ovines, swines, canines, felines, rodents, and primates, of which the preferred Mammals are humans, dogs or pigs.
  • the COVID-19 described in this application is a disease caused by SARS-CoV-2.
  • the pharmaceutical composition described in this application further comprises a pharmaceutically acceptable carrier or excipient.
  • the pharmaceutical composition may be a solid preparation, an injection, an inhalation preparation, a spray, a liquid preparation, or a compound preparation.
  • composition described in this application can be prepared into various forms according to different administration routes, such as oral tablets, capsules, powders, oral liquids, injections and transdermal preparations.
  • pharmaceutically acceptable carriers include diluents, fillers, disintegrants, wetting agents, lubricants, coloring agents, flavoring agents or other conventional additives.
  • Typical pharmaceutically acceptable carriers include, for example, microcrystalline cellulose, starch, crospovidone, povidone, polyvinylpyrrolidone, maltitol, citric acid, sodium lauryl sulfonate or magnesium stearate, etc. .
  • the pharmaceutical composition can be administered in any of the following ways: oral, spray inhalation, rectal, nasal, buccal, vaginal, topical, parenteral, such as subcutaneous, intravenous, intramuscular, Intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or medication with the aid of an explanted reservoir.
  • oral, intraperitoneal or intravenous administration is preferred.
  • the compound represented by the general formula I or its geometric isomer, its pharmaceutically acceptable salt, its solvate and/or its hydrate can be made into any orally acceptable preparation form, including But not limited to tablets, capsules, aqueous solutions or suspensions.
  • the carriers commonly used in tablets include lactose and corn starch, and lubricants such as magnesium stearate can also be added.
  • Diluents commonly used in capsule preparations include lactose and dried corn starch.
  • Aqueous suspension formulations usually mix the active ingredients with suitable emulsifiers and suspending agents. If necessary, some sweeteners, fragrances or coloring agents can be added to the above oral preparations.
  • the compound represented by the general formula I or its geometric isomer, its pharmaceutically acceptable salt, its solvate and/or its hydrate can generally be made into the form of suppositories, which can be used to form a suppository. It is prepared by mixing with a suitable non-irritating excipient.
  • the excipient presents a solid state at room temperature, but melts at the rectal temperature to release the drug.
  • excipients include cocoa butter, beeswax and polyethylene glycol.
  • topical medication especially for the treatment of affected surfaces or organs that are easily reached by topical application, such as eye, skin or lower intestinal neurological diseases
  • the compound represented by the general formula I or its geometric isomers, and its pharmaceuticals Acceptable salts, their solvates and/or their hydrates can be made into different topical formulations according to different affected surfaces or organs, as detailed below:
  • the compound represented by the general formula I or its geometric isomer, its pharmaceutically acceptable salt, its solvate and/or its hydrate can be formulated into a micronized suspension or solution
  • the carrier used is isotonic sterile saline with a certain pH, which may or may not be added with preservatives such as chlorinated benzyl alkanolate.
  • the compound can also be made into an ointment form such as petrolatum ointment.
  • the compound represented by the general formula I or its geometric isomer, its pharmaceutically acceptable salt, its solvate and/or its hydrate can be made into an appropriate ointment, lotion or cream A dosage form in which the active ingredient is suspended or dissolved in one or more carriers.
  • the carriers that can be used for the ointment here include, but are not limited to: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; the carriers that can be used for lotions or creams include but are not limited to: Mineral oil, sorbitan monostearate, Tween 60, cetyl ester wax, hexadecenyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
  • the compound represented by the general formula I or its geometric isomer, its pharmaceutically acceptable salt, its solvate and/or its hydrate can be made into the rectal suppository formulation as described above Or a suitable enema preparation form, and a topical transdermal patch can also be used.
  • the compound represented by the general formula I or its geometric isomer, its pharmaceutically acceptable salt, its solvate and/or its hydrate can also be administered in the form of a sterile injection preparation, including sterile injection water or oil suspension Solution, or sterile injection solution.
  • a sterile injection preparation including sterile injection water or oil suspension Solution, or sterile injection solution.
  • usable carriers and solvents include water, Ringer's solution and isotonic sodium chloride solution.
  • sterilized non-volatile oils can also be used as solvents or suspension media, such as monoglycerides or diglycerides.
  • therapeutically effective amount refers to an amount that is sufficient to treat or prevent the patient's disease but is low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of reasonable medical judgment .
  • the therapeutically effective amount of the compound will depend on the specific compound selected (for example, considering the potency, effectiveness and half-life of the compound), the route of administration selected, the disease to be treated, the severity of the disease to be treated, and the patient's condition to be treated. Factors such as age, size, weight, and physical disease, the medical history of the patient being treated, the duration of treatment, the nature of concurrent therapy, the desired therapeutic effect and other factors have changed, but they can still be routinely determined by those skilled in the art.
  • the specific dosage and method of use of the compound represented by the general formula I or its geometric isomers, its pharmaceutically acceptable salts, its solvates and/or its hydrates for different patients are determined by many Factors include the patient’s age, weight, gender, natural health status, nutritional status, active strength of the drug, taking time, metabolic rate, severity of the disease, and the subjective judgment of the physician.
  • the preferred dosage here is between 0.001-1000 mg/kg body weight/day.
  • Figure 3 shows the quantitative RT-PCR detection results of tetrandrine inhibiting SARS-CoV-2 virus infection on Vero cells.
  • Tetrandrine is from Zhejiang Jinhua Kangenbei Pharmaceutical Co., Ltd., batch number YG1910005, and the content is 99.3%.
  • the reference drug, chloroquine phosphate was purchased from Sigma Company, article number C6628; Redecivir was purchased from MedChemExpresss Company, article number HY-104077.
  • Tetrandrine was prepared with DMSO to prepare a 10 mM mother solution, Redecivir was prepared with DMSO to prepare a 100 mM mother solution, and chloroquine phosphate was prepared with PBS to prepare a 100 mM mother solution.
  • the MEM culture medium containing 2% fetal bovine serum was diluted to the concentration required for the experiment.
  • Vero-E6 cells purchased from ATCC, catalog number 1586.
  • MEM dry powder purchased from GIBCO in the United States, item number 10370021
  • Fetal Bovine Serum (FBS) purchased from GIBCO in the U.S., item number 16000044
  • sodium bicarbonate purchased from Sinopharm Group
  • penicillin, streptomycin and kana Mycin All were purchased from North China Pharmaceutical Factory
  • PBS was purchased from Gibco Company, catalog number C10010500BT.
  • the cell culture medium is: MEM culture medium, containing 10% fetal bovine serum per 100ml, 100U/ml each of penicillin, streptomycin and kanamycin, and NaHCO 3 5%.
  • Cell digestion solution 0.25% pancreatin, prepared with Hanks solution, 0.02% EDTA.
  • Vero E6 cell culture add 0.1ml 0.25% trypsin, 5ml 0.02% EDTA, digest at 37°C for 5 minutes in a culture flask full of cells, discard the digestion solution, add cell culture solution and pipette, passaging 1:3, 3
  • the sky is prepared to make 100,000 cells per milliliter, inoculate a 96-well cell culture plate, 0.1ml per well, 37°C, 5% CO 2 culture for 24 hours, the cells grow into a monolayer and then conduct the experiment.
  • Cell viability (%) (A (drug treatment group) -A (blank control) )/(A (negative control) -A (blank control) ) ⁇ 100%
  • MEM culture medium containing 2% FBS
  • tetrandrine, remdesivir and chloroquine phosphate of corresponding concentrations to make the drug in the hole final
  • concentrations are as shown in Table 4 to Table 6, and then placed in a 37°C, 5% CO 2 incubator to continue culturing for 48 hours.
  • the cell control group is added with a final concentration of 0.3% DMSO containing 2% FBS MEM medium, or 0.5% PBS containing 2% FBS MEM medium.
  • the kit (TaKaRa MiniBEST Viral RNA/DNA Extraction Kit, item number 9766) produced by TaKaRa was used for RNA extraction. Unless otherwise specified, the consumables and reagents involved in the following RNA extraction steps are part of the kit. The following extraction steps are recommended steps in the kit instructions.
  • nuclease-free EP tube purchased from Axygen, item number mct-150-c
  • 321 ⁇ L of lysate 100 ⁇ L PBS, 200 ⁇ L buffer VGB, 2 ⁇ L proteinase to each well K, 1 ⁇ L carrier RNA
  • mix well and digest at 56°C for 15min;
  • the reverse transcription kit (PrimeScript TM RT reagent Kit with gDNA Eraser, article number RR047Q) produced by TaKaRa was used for RNA reverse transcription. Proceed as follows:
  • RNA samples of each experimental group and take 3 ⁇ L of RNA for reverse transcription.
  • Fluorescence quantitative PCR was used to detect the number of copies per milliliter of the original virus solution.
  • RBD-qR CTCAAGTGTCTGTGGATCACG
  • Cycle parameters 95°C for 15 seconds, 54°C for 15 seconds, 72°C for 30 seconds. A total of 40 cycles.
  • test compound can effectively inhibit the replication of the viral genome in the supernatant of SARS-CoV-2 infection at different concentrations (Table 4-6 and Figure 1).
  • Tetrandrine (reference substances chloroquine phosphate and remdesivir) has good safety to test cells in vitro at different test concentrations. Tetrandrine, remdesivir and chloroquine phosphate have a significant inhibitory effect on the SARS-CoV-2 virus isolate nCoV-2019BetaCoV/Wuhan/WIV04/2019.
  • MOI of the virus infection When the MOI of the virus infection is 0.01, the EC 50 values are respectively 1.71 ⁇ M, 0.30 ⁇ M and 1.54 ⁇ M, the corresponding therapeutic index is 14.33, 534.33, 60.34, respectively.
  • the MOI of virus infection is 0.05, the EC 50 values are 2.88 ⁇ M, 0.59 ⁇ M, and 3.77 ⁇ M, and the corresponding therapeutic indexes are 8.51, 271.69, and 24.49, respectively.
  • Tetrandrine, chloroquine phosphate and remdesivir have inhibitory activity against SARS-CoV-2 virus in vitro.
  • Example 1 The experimental materials and methods are the same as in Example 1. The specific difference is that in the antiviral experiment, the drug treatment step in Example 1 is to first add 100 ⁇ L of MEM culture medium (containing 2% FBS) containing the corresponding concentration of the drug to the cell plate to pretreat the cells 1 hour, and the drug treatment step in this embodiment does not require drug pretreatment of the cells.
  • MEM culture medium containing 2% FBS
  • the specific steps are as follows:
  • the virus culture medium was discarded, the uninfected residual virus was washed with PBS, and the MEM culture medium (containing 2% FBS) containing the corresponding concentration of tetrandrine, remdesivir and chloroquine phosphate was added to make the medicine in the well
  • the final concentration is the same as the drug concentration shown in Tables 4, 5, and 6 of Example 1, and then placed in a 37°C, 5% CO 2 incubator for 48 hours.
  • the cell control group is added with a final concentration of 0.3% DMSO containing 2% FBS.
  • RNA extraction, RNA reverse transcription and Real-time PCR methods are the same as those in Example 1.
  • Tetrandrine, chloroquine phosphate and remdesivir have inhibitory activity against SARS-Cov-2 virus after the virus infects cells.
  • Example 3 tetrandrine prime half of the maximal effective concentration of the SARS-CoV-2 (EC 50)
  • a nucleic acid quantification method was used to determine the EC 50 of the drug. The details are as follows: one day in advance, Vero cells were seeded into a 48-well plate at a concentration of about 10,000 cells/well. The drug tetrandrine was formulated into the final concentration of 20, 10, 5, 2.5, 1.25, 0.625 and 0.3125 ⁇ M in DMEM medium containing 2% FBS (purchased from Gibco, article number 16000044), added to the cells, and placed 37 °C 5% CO 2 incubator pretreatment for 1 hour.
  • the SARS-CoV-2 virus (nCoV-2019BetaCoV/Wuhan/WIV04/2019 isolate, GISAID accession number EPI_ISL_402124, isolated and subcultured by Wuhan Institute of Virology, Chinese Academy of Sciences) was diluted with DMEM medium containing 2% FBS, and then Add to the corresponding wells, make the viral load 100 TCID 50 , absorb and incubate at 37°C for 2 hours, discard the virus solution, and add different concentrations of hexetidine to each well (200 ⁇ l/well), so that each well The final concentration of A is 20, 10, 5, 2.5, 1.25, 0.625, and 0.3125 ⁇ M, 3 replicate wells are set for each drug concentration, virus control and normal cell control are set up, and placed in a 37°C 5% CO 2 incubator Culture and observe cytopathic (CPE) every day. Two days after infection, 50 ⁇ l of cell supernatant was taken from each well to extract nucleic acid, and quantitative RT-PCR was
  • RNA extraction RNA reverse transcription and RT-PCR are the same as in Example 1.
  • infection rate (%) drug group RNA copy number/virus control RNA copy number ⁇ 100%) to calculate the virus infection rate (%) under different concentrations of drug treatment, and use Graphpad Prism 7 software to simulate the data Combined analysis, the fitting result is shown in Figure 3, and the EC 50 is calculated.

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Abstract

用于预防和/或治疗与SARS-CoV-2有关的肺部疾病或症状或无症状或有症状的SARS-CoV-2感染的通式I所示的化合物、其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，及通式I所示的化合物、其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物在制备用于预防和/或治疗与SARS-CoV-2有关的肺部疾病或症状，或无症状或有症状的SARS-CoV-2感染的产品中用途。

Description

二甲基小檗胺类化合物在抑制SARS-CoV-2中的用途
本申请是以CN申请号为202010140356.0，申请日为2020年3月3日的申请为基础，并主张其优先权，该CN申请的公开内容在此作为整体引入本申请中。
技术领域
本申请属于化学药物领域，具体涉及下面式I所示二甲基小檗胺类化合物，其几何异构体或其药学上可接受的盐和/或两者溶剂化物和/或两者水合物，及含上述化合物的药物组合物，用于治疗SARS-CoV-2感染方面的用途。
Figure PCTCN2020101581-appb-000001
背景技术
式I所示二甲基小檗胺类化合物，化学名称为6,6',7,12-四甲氧基-2,2'-二甲基小檗胺，是从防己科植物粉防己(Stephania tetrandra)中提取的双苄基异喹啉类生物碱。式(I)化合物又称汉防己甲素(Tetrandrine)，别名粉防己碱、汉防己碱，为抗高血压药，临床上主要用于治疗轻、中度高血压，亦可用于高血压危象。此外，也可用于风湿痛、关节痛、神经痛。与小剂量放射合并用于肺癌；亦用于单纯硅肺I、II、III期及各期煤硅肺。
冠状病毒(Coronavirus)是有包膜的不分节段的单股正链RNA病毒，具有广泛的动物宿主。来源于动物传染病的SARS冠状病毒和MERS冠状病毒可引起人类的死亡，并分别在2002年和2012年爆发(Zaki et al.,2012；Zhong et al.,2003；Cui et al.,2019)。2019新型冠状病毒(2019-nCoV)是以前从未在人类中发现的冠状病毒新毒株。2020年2月11日，国际病毒分类委员会(ICTV)宣布，2019新型冠状病毒(2019-nCoV)的正式分类名为严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2，SARS-CoV-2)。同日，世界卫生组织(WHO)宣布，由这一病毒导致的疾病的正式名称为COVID-19。COVID-19患者的临床表现 为，90％以上的患者表现为发热，80％的患者表现为干咳，20％的患者表现为呼吸急促，15％的患者表现为呼吸困难，而最重要的表现是白细胞和淋巴细胞的减少，患者肺部均有不同程度的器官损伤，具有发展成为肺纤维化的潜在风险。该病毒具有很强的传染性，全球范围内的多个国家，包括日本、韩国、意大利和美国等，都出现了SARS-CoV-2感染事件，并有不断扩大的趋势。前期的研究发现，瑞德西韦(remdesivir)和磷酸氯喹(chloroquine)等药物可以很好的抑制SARS-CoV-2病毒的复制(Wang et al.,2020)，也已经在临床上开展了相关研究，相关的疫苗也加紧进行研制。然而，目前尚无一种药物或疫苗获得批准广泛应用。
因此，研发能够有效抑制SARS-CoV-2的产品仍是十分紧迫和必要的。
发明内容
本发明人在研究中发现汉防己甲素能够有效抑制SARS-CoV-2病毒，预防或治疗肺纤维化，从而为有效预防和/或治疗SARS-CoV-2病毒引发的肺部疾病或症状提供了新的途径和选择。本申请基于以上研究现已完成。
本申请涉及用于预防和/或治疗与SARS-CoV-2有关的肺部疾病或症状或无症状或有症状的SARS-CoV-2感染的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，
Figure PCTCN2020101581-appb-000002
本申请还涉及用于预防和/或治疗与SARS-CoV-2有关且伴有高血压的肺部疾病，或症状或伴有高血压的无症状或有症状的SARS-CoV-2感染的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，
Figure PCTCN2020101581-appb-000003
本申请还涉及用于预防和/或治疗COVID-19或伴有高血压的COVID-19，或作为SARS-CoV-2抑制剂，或用于抑制SARS-CoV-2在细胞(例如哺乳动物细胞)中复制或繁殖的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，
Figure PCTCN2020101581-appb-000004
本申请进一步涉及用于预防和/或治疗冠状病毒引起的疾病或感染的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
本发明进一步涉及用于预防和/或治疗冠状病毒引起的并伴有高血压的疾病或感染的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于预防和/或治疗冠状病毒引起的疾病或感染或冠状病毒引起的并伴有高血压的疾病或感染。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/ 或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于预防和/或治疗与冠状病毒有关且伴有高血压的肺部疾病或症状。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于抑制冠状病毒在细胞(例如哺乳动物细胞)中复制或繁殖。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其作为冠状病毒抑制剂。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗冠状病毒引起的疾病或感染的产品中用途。
本发明进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗冠状病毒引起的并伴有高血压的疾病或感染的产品中用途。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗与冠状病毒有关且伴有高血压的肺部疾病或症状的产品中用途。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于抑制冠状病毒在细胞(例如哺乳动物细胞)中复制或繁殖的产品中的用途。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备作为冠状病毒抑制剂的产品中的用途。
本申请进一步涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗与SARS-CoV-2有关的肺部疾病或症状，或无症状或有症状的SARS-CoV-2感染的产品中用途。
本申请还涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗与SARS-CoV-2有关且伴有高血压的肺部疾病或症状，或伴有高血压的无症状或有症状的SARS-CoV-2感染的产品中用途。
本申请还涉及通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗COVID-19或伴有高血压的COVID-19的产品中的用途，或在制备作为SARS-CoV-2抑制剂的产品中的用途，或在制备用于抑制SARS-CoV-2在细胞(例如哺乳动物细胞)中复制或繁殖的产品中的用途。
本申请进一步涉及预防和/或治疗冠状病毒引起的疾病或感染或冠状病毒引起的并伴有高血压的疾病或感染的方法，其包括给有需要的宿主施用将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
本申请进一步涉及预防和/或治疗与冠状病毒有关的肺部疾病或症状或与冠状病毒有关且伴有高血压的肺部疾病或症状的方法，其包括给有需要的宿主施用将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
本申请进一步涉及在有需要的宿主中抑制冠状病毒复制或繁殖的方法，其包括给有需要的宿主施用将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
本申请还涉及预防和/或治疗与SARS-CoV-2有关的肺部疾病或症状或无症状或有症状的SARS-CoV-2感染的方法，其包括将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物给予与患有SARS-CoV-2有关的肺部疾病或症状或患有无症状或有症状的SARS-CoV-2感染的宿主。
本申请还涉及预防和/或治疗与SARS-CoV-2有关且伴有高血压的肺部疾病或症状或伴有高血压的无症状或有症状的SARS-CoV-2感染的方法，其包括将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物给予与SARS-CoV-2有关且患有高血压的肺部疾病或症状或患有高血压的无症状或有症状的SARS-CoV-2感染的宿主。
本申请还涉及在有需要的宿主中治疗和/或预防COVID-19或伴有高血压的COVID-19的方法或者在有需要的宿主中抑制SARS-CoV-2复制或繁殖的方法，该方法包括给有需要的宿主施用治疗和/或预防有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
在某些实施方案中，本申请所述冠状病毒为SARS-CoV-2。
在某些实施方案中，本申请所述冠状病毒为SARS-CoV-2。
在某些实施方案中，本申请所述的冠状病毒引起的疾病为COVID-19。
在某些实施方案中，本申请所述的冠状病毒引起的并伴有高血压的疾病为伴有高血压的COVID-19。
在某些实施方案中，本申请所述的与冠状病毒有关的肺部疾病或症状为SARS-CoV-2有关的肺部疾病或症状。
在某些实施方案中，本申请所述与冠状病毒有关且伴有高血压的肺部疾病或症状为SARS-CoV-2有关且伴有高血压的肺部疾病或症状。
在某些实施方案中，本申请所述的冠状病毒引起的感染为无症状或有症状的SARS-CoV-2感染。
在某些实施方案中，本申请所述的冠状病毒引起的并伴有高血压的感染为伴有高血压的无症状或有症状的SARS-CoV-2感染。
在某些实施方案中，本申请所述产品为人用药品或动物用兽药。
在某些实施方案中，本申请所述产品为固体制剂、注射剂、喷剂、液体制剂、吸入制剂或复方制剂。
在某些实施方案中，本申请所述宿主为哺乳动物。
在某些实施方案中，本申请所述哺乳动物包括牛科动物、马科动物、羊科动物、猪科动物、犬科动物、猫科动物、啮齿类动物、灵长类动物，其中优选的哺乳动物是人，狗或猪。
在某些实施方案中，本申请所述COVID-19是由SARS-CoV-2导致的疾病。
在某些实施方案中，本申请所述的药物组合物还包含药学上可接受的载体或辅料。具体地，所述药物组合物可以为固体制剂、注射剂、吸入制剂、喷剂、液体制剂、或复方制剂。
本申请所述药物组合物可以根据不同给药途径而制备成各种形式，例如口服片剂，胶囊，粉剂，口服液，注射剂和透皮制剂。根据常规的药物上的惯例，药学上可接受的载体包括稀释剂、填充剂、崩解剂、润湿剂、润滑剂、着色剂、调味剂或其它常规添加剂。典型的药学上可接受的载体包括例如微晶纤维素、淀粉、交连聚维酮、聚维酮、聚乙烯吡咯烷酮、麦芽糖醇，柠檬酸，十二烷基磺酸钠或硬脂酸镁等。
根据申请，所述的药物组合物可以以下面的任意方式施用：口服、喷雾吸入、直肠用药、鼻腔用药、颊部用药、阴道用药、局部用药、非肠道用药如皮下、静脉、肌内、腹膜内、鞘内、心室内、胸骨内和颅内注射或输入、或借助一种外植储器用药。其中优选口服、腹膜内或静脉内用药方式。
当口服用药时，所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物可制成任意口服可接受的制剂形式，包括但不限于片剂、胶囊、水溶液或水悬浮液。其中，片剂一般使用的载体包括乳糖和玉米淀粉，另外也可加入润滑剂如硬质酸镁。胶囊制剂一般使用的稀释剂包括乳糖和干燥玉米淀粉。水悬浮液制剂则通常是将活性成分与适宜的乳化剂和悬浮剂混合使用。如果需要，以上口服制剂形式中还可加入一些甜味剂、芳香剂或着色剂。
当直肠用药时，所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物一般可制成栓剂的形式，其通过将药物与一种适宜的非刺激性赋形剂混合而制得。该赋形剂在室温下呈现固体状态，而在直肠温度下熔化释出药物。该类赋形剂包括可可脂、蜂蜡和聚乙二醇。
当局部用药时，特别是治疗局部外敷容易达到的患面或器官，如眼睛、皮肤或下肠道神经性疾病时，所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物可根据不同的患面或器官制成不同的局部用药制剂形式，具体说明如下：
当眼部局部施用时，所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物可配制成一种微粉化悬浮液或溶液的制剂形式，所使用载体为等渗的一定pH的无菌盐水，其中可加入也可不加防腐剂如氯化苄基烷醇盐。此外对于眼用，也可将化合物制成膏剂形式如凡士林膏。
当皮肤局部施用时，所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物可制成适当的软膏、洗剂或霜剂制剂形式，其中活性成分悬浮或溶解于一种或多种载体中。这里软膏即可使用的载体包括但不限于：矿物油、液体凡士林、白凡士林、丙二醇、聚氧化乙烯、聚氧化丙烯、乳化蜡和水；洗剂或霜剂可使用的载体包括但不限于：矿物油、脱水山梨糖醇单硬脂酸酯、吐温60、十六烷酯蜡、十六碳烯芳醇、2-辛基十二烷醇、苄醇和水。
当下肠道局部施用时，所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物可制成如上所述的直肠栓剂制剂或适宜的灌肠制剂形式，另外也可使用局部透皮贴剂。
所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物还可以无菌注射制剂形式用药，包括无菌注射水或油悬浮液，或无菌注射溶液。其中，可使用的载体和溶剂包括水，林格氏溶液和等渗氯化钠溶液。另外，灭菌的非挥发油也可用作溶剂或悬浮介质，如单甘油酯或二甘油酯。
上述各种剂型的药物均可以按照药学领域的常规方法制备。
如本文所述的，“治疗有效量”或“预防有效量”是指在合理的医学判断范围内，足以治疗或预防患者疾病但足够低地避免严重副作用(在合理的利益/风险比)的量。化合物的治疗有效量将根据所选择的具体化合物(例如考虑化合物的效力、有效性和半衰期)、所选择的给药途径、所治疗的疾病、所治疗的疾病的严重性、所治疗的患者的年龄、大小、体重和身体疾病、所治疗的患者的医疗史、治疗持续时间、并行疗法的性质、所需的治疗效果等因素发生变化，但仍可以由本领域技术人员常规确定。
另外需要指出，所述通式I所示的化合物或其几何异构体、其药物上可接受的盐、其溶剂化物和/或其水合物针对不同患者的特定使用剂量和使用方法决定于诸多因素， 包括患者的年龄，体重，性别，自然健康状况，营养状况，药物的活性强度，服用时间，代谢速率，病症的严重程度以及诊治医师的主观判断。这里优选使用剂量介于0.001-1000mg/kg体重/天。
附图说明
图1为实施例1中，汉防己甲素、瑞得西韦和磷酸氯喹抑制SARS-CoV-2的量效曲线，其中蓝色点代表药物在不同浓度下的细胞毒性百分率；红色曲线为药物在不同浓度下抑制病毒产量百分率，上图病毒感染复数MOI＝0.01；下图病毒感染复数为MOI＝0.05。
图2为实施例2中，汉防己甲素、瑞得西韦和磷酸氯喹抑制SARS-CoV-2的量效曲线，其中蓝色点代表药物在不同浓度下的细胞毒性百分率；红色曲线为药物在不同浓度下抑制病毒产量百分率，上图病毒感染复数MOI＝0.01；下图病毒感染复数为MOI＝0.05。
图3为汉防己甲素在Vero细胞上抑制SARS-CoV-2病毒感染的定量RT-PCR检测结果。
具体实施方式
下面结合本申请的具体实施例来进一步说明本申请的实质性内容，应理解，以下实施例仅用于说明本申请，但并不以此来限定本申请的保护范围。下面实施例中未注明具体条件者，按照常规条件或制造商建议的进行。所用药品或试剂未注明生产厂商者，均为可以通过市购获得的常规产品。
虽然以下实施例中所使用的许多材料和操作方法是本领域公知的，但是本申请仍然在此作尽可能详细描述。本领域技术人员清楚，如果未特别说明，下面实施例中所用的材料和操作方法是本领域公知的。
实施例1汉防己甲素对SARS-CoV-2的药效学研究1
1.材料和方法
1.1药物：汉防己甲素来源于浙江金华康恩贝制药有限公司，批号YG1910005，含量为99.3％。对照药品磷酸氯喹购置于Sigma公司，货号C6628；瑞得西韦购自MedChemExpresss公司，货号HY-104077。汉防己甲素用DMSO配成10mM母液，瑞得西韦用DMSO配成100mM母液，磷酸氯喹用PBS配成100mM母液。实验时用 含2％胎牛血清的MEM培养液稀释成实验所需的浓度。
1.2细胞：Vero-E6细胞，购自ATCC，货号1586。
1.3病毒：SARS-CoV-2病毒
(nCoV-2019BetaCoV/Wuhan/WIV04/2019分离株，GISAID登录号EPI_ISL_402124)由中国科学院武汉病毒研究所分离传代培养。实验时，用含2％胎牛血清的MEM培养液将病毒稀释至实验所需浓度。
1.4.试剂、实验用品及仪器：
1.4.1试剂：MEM干粉，购自美国GIBCO公司，货号10370021；胎牛血清(FBS)，购自美国GIBCO公司，货号16000044；碳酸氢钠，购自国药集团；青霉素、链霉素和卡那霉素：均购自华北制药厂；PBS购自Gibco公司，货号C10010500BT。
1.4.2实验用品及仪器：培养瓶，购自美国Corning公司；培养板96孔板，购自美国Corning公司；二氧化碳孵箱，购自美国Thermo公司；
1.4.3细胞培养液及试剂配制：
细胞培养液为：MEM培养液，每100ml含胎牛血清10％，青霉素，链霉素和卡那霉素各100U/ml，NaHCO 3 5％。
细胞消化液：0.25％胰酶，用Hanks液配制，0.02％EDTA。
1.5.实验方法：
1.5.1Vero E6细胞培养：在长满细胞的培养瓶内加0.25％胰酶0.1ml，0.02％EDTA 5ml，37℃消化5分钟，废弃消化液，加细胞培养液吹打，1:3传代，3天长满，配制成每毫升10万个细胞，接种96孔细胞培养板，每孔0.1ml，37℃，5％CO 2培养24小时，细胞长成单层后进行实验。
1.5.2药物对细胞毒性实验：
药物对细胞毒性的检测利用CellTiter-Glo试剂盒(Promega)测定。具体步骤如下：
①96孔板中接种1×10 4个Vero-E6细胞，37℃培养8小时。
②将药物的母液用含2％FBS的MEM培养液稀释到所需浓度，弃96孔板中原培养液，取100μL含药物的MEM培养液加入到对应的孔中，使孔中药物最终浓度分别如表1至表3中所示，每个浓度做三个复孔。同时设置阴性对照(细胞孔中加DMSO或PBS，和含2％FBS的MEM培养液，而不加药物)和空白对照(不含细胞，加DMSO和含2％FBS的MEM培养液)。加药完毕，将细胞置于37℃培养24小时。
③向待测孔中加入100μL CellTiter-Glo溶液(Promega)，震荡混匀2min使细胞充分裂解，室温孵育10min后，在酶标仪(购自Molecular Devices公司，型号SpectraMax M5)上读取化学发光信号OD450，将读数带入以下公式，计算细胞活性：
细胞活性(％)＝(A (药物处理组)-A (空白对照))/(A (阴性对照)-A (空白对照))×100％
其中A为酶标仪读数。
1.5.3抗病毒实验：
①药物处理
将Vero E6细胞接种到48孔板中，每孔约1×10 5个细胞，待第二天实验。先将100μL含相应浓度药物的MEM培养液(含有2％FBS)加入细胞板中，预处理细胞1小时，然后加入20μL稀释的病毒(病毒量为1000TCID 50,即MOI＝0.01，以及病毒量为5000TCID 50,即MOI＝0.05)，置于培养箱孵育1小时。然后弃病毒培养液，用PBS洗去未感染的残留病毒，再加入含相应浓度的汉防己甲素、瑞得西韦和磷酸氯喹的MEM培养液(含有2％FBS)，使孔中药物最终浓度分别如表4至表6中所示，然后放入37℃、5％CO 2孵箱继续培养48h，细胞对照组加入终浓度0.3％DMSO的含有2％FBS的MEM培养液，或0.5％PBS的含有2％FBS的MEM培养液。
②RNA提取
采用TaKaRa公司生产的试剂盒(TaKaRa MiniBEST Viral RNA/DNA Extraction Kit，货号9766)进行RNA提取。如果没有特别指出，下述RNA提取步骤中所涉及的耗材及试剂均为试剂盒的组成部分。下述提取步骤均为试剂盒说明书所推荐的步骤。
1)取受试培养板的上清液100μL，加入无核酸酶EP管(购自Axygen，货号mct-150-c)中，然后每孔加入321μL裂解液(100μL PBS,200μL buffer VGB,2μL proteinase K,1μL carrier RNA)，混匀后置于56℃消化15min；
2)向1)中所得混合液加200μL无水乙醇，混匀；
3)将上述2)中所得混合液转入无RNA酶的离心柱中，12000rpm离心15s，弃废液；
4)加入500μL Buffer RW1，12000rpm离心15s清洗离心柱，弃废液；
5)加入650μL Buffer RW2，12000rpm离心15s清洗离心柱，弃废液；
6)加入650μL Buffer RW2，12000rpm离心2min清洗离心柱，弃废液，然后将离心柱整体转移至7)中的新的无RNA酶的2ml收集管中；
7)换新的无RNA酶的2ml收集管，12000rpm离心1min，干燥离心柱，然后将 离心柱整体转移至8)中的1.5ml收集管中；
8)换上新的1.5ml收集管，放入7)中干燥的离心柱，向每个离心柱中加入30μl不含RNA酶的水，12000rpm离心2min，洗脱液即含有相应的RNA。
③RNA反转录
实验采用TaKaRa公司生产的反转录试剂盒(PrimeScript TM RT reagent Kit with gDNA Eraser，货号RR047Q)进行RNA反转录。步骤如下：
1)进行gDNA去除：收集各实验组RNA样品，分别取3μL RNA进行反转录。首先，向各实验组RNA中加入2μl 5×gDNA Eraser Buffer，用RNase Free水补足反应体系至10μl，充分混匀，42℃水浴2min去除样品中可能存在的g DNA；
2)进行逆转录：向1)中所得样品中加入适量的酶和引物Mix及反应缓冲液，用RNase Free水补足体积至20μl，37℃水浴反应15min，之后投入85℃水中5sec，既可转录得到cDNA。
④Real-time PCR。
采用荧光定量PCR检测原病毒液每毫升所含拷贝数。
采用TB Green Premix(Takara，Cat#RR820A)混好反应体系，在StepOne Plus Real-time PCR仪(品牌：ABI)进行扩增反应和读数。计算原病毒液每毫升所含拷贝数。步骤如下：
1)首先建立标准品：将质粒pMT-RBD稀释成5×10 8copies/μL,5×10 7copies/μL,5×10 6copies/μL,5×10 5copies/μL,5×10 4copies/μL,5×10 3copies/μL,5×10 2copies/μL。取2μL标准品或cDNA模板用于qPCR反应。；
2)实验过程中所用引物序列如下(均为5’-3’方向表示)：
RBD-qF:CAATGGTTTAACAGGCACAGG
RBD-qR:CTCAAGTGTCTGTGGATCACG
3)反应程序如下：
预变性：95℃ 5分钟；
循环参数：95℃ 15秒，54℃ 15秒，72℃ 30秒。共40个循环。
2.结果
2.1.汉防己甲素、磷酸氯喹和瑞得西韦对Vero E6细胞的毒性
细胞毒性结果显示，在所有受试浓度下，受试化合物的处理均未改变细胞活力，即受试化合物在所有浓度下对细胞均无毒性作用(表1-3)。
表1.受试化合物汉防己甲素的细胞毒性实验
Figure PCTCN2020101581-appb-000005
表2.受试化合物瑞得西韦的细胞毒性实验
Figure PCTCN2020101581-appb-000006
表3.受试化合物磷酸氯喹的细胞毒性实验
Figure PCTCN2020101581-appb-000007
2.2.汉防己甲素、磷酸氯喹和瑞得西韦的抗病毒活性
病毒增殖抑制实验的结果显示，受试化合物在不同的浓度下，均能够有效抑制SARS-CoV-2感染上清中病毒基因组的复制(表4-6和图1)。
表4.受试化合物汉防己甲素的抗病毒实验
Figure PCTCN2020101581-appb-000008
表5.受试化合物瑞得西韦的抗病毒实验
Figure PCTCN2020101581-appb-000009
表6.受试化合物磷酸氯喹的抗病毒实验
Figure PCTCN2020101581-appb-000010
经计算，在病毒感染复数MOI＝0.01的条件下：
汉防己甲素EC 50＝1.71μM，CC 50＝24.51μM，SI＝14.33
瑞得西韦EC 50＝0.30μM，CC 50＝160.30μM，SI＝534.33
磷酸氯喹EC 50＝1.54μM，CC 50＝92.93μM，SI＝60.34
在病毒感染复数MOI＝0.05的条件下：
汉防己甲素EC 50＝2.88μM，CC 50＝24.51μM，SI＝8.51
瑞得西韦EC 50＝0.59μM，CC 50＝160.30μM，SI＝271.69
磷酸氯喹EC 50＝3.77μM，CC 50＝92.93μM，SI＝24.49
3.结论
汉防己甲素(对照品磷酸氯喹和瑞得西韦)在不同受试浓度下在体外水平对受试细胞具有良好的安全性。汉防己甲素、瑞得西韦和磷酸氯喹对SARS-CoV-2病毒分离株nCoV-2019BetaCoV/Wuhan/WIV04/2019有明显的抑制作用，在病毒感染的MOI为0.01时，EC 50值分别为1.71μM，0.30μM和1.54μM，相应的治疗指数分别为14.33，534.33，60.34。在病毒感染的MOI为0.05时，EC 50值分别为2.88μM，0.59μM和3.77μM，相应的治疗指数分别为8.51，271.69，24.49。
汉防己甲素、磷酸氯喹和瑞得西韦在体外水平对SARS-CoV-2病毒具有抑制活性。
实施例2汉防己甲素对SARS-CoV-2的药效学研究2
实验材料、方法同实施例1，具体区别在于抗病毒实验中，实施例1中药物处理步骤是先将100μL含相应浓度药物的MEM培养液(含2％FBS)加入细胞板中，预处理细胞1小时，而本实施例中的药物处理步骤不用药物预处理细胞，具体步骤如下：
将Vero E6细胞接种到48孔板中，每孔约1×10 5个细胞，然后加入20μL稀释 的病毒(病毒量为1000TCID 50,即MOI＝0.01，以及病毒量为5000TCID 50,即MOI＝0.05)，置于培养箱孵育1小时。1h后弃病毒培养液，用PBS洗去未感染的残留病毒，再加入含相应浓度的汉防己甲素、瑞得西韦和磷酸氯喹的MEM培养液(含2％FBS)，使孔中药物最终浓度同实施例1表4、5、6所示的各个药物浓度，然后放入37℃、5％CO 2孵箱继续培养48h，细胞对照组加入终浓度0.3％DMSO的含有2％FBS的细胞培养液，或0.5％PBS的含有2％FBS的细胞培养液。
后续的RNA提取、RNA反转录和Real-time PCR方法均与实施例1相同。
经计算，在病毒感染复数MOI＝0.01的条件下：
汉防己甲素EC 50＝1.61μM，CC 50＝24.51μM，SI＝15.22
瑞得西韦EC 50＝0.23μM，CC 50＝160.30μM，SI＝696.96
磷酸氯喹EC 50＝4.52μM，CC 50＝92.93μM，SI＝20.56
在病毒感染复数MOI＝0.05的条件下：
汉防己甲素EC 50＝1.95μM，CC 50＝24.51μM，SI＝12.51
瑞得西韦EC 50＝0.62μM，CC 50＝160.30μM，SI＝258.55
磷酸氯喹EC 50＝3.63μM，CC 50＝92.93μM，SI＝25.60
结论：
抗病毒实验结果表明，汉防己甲素、瑞得西韦和磷酸氯喹对SARS-Cov-2病毒分离株nCoV-2019BetaCoV/Wuhan/WIV04/2019有明显的抑制作用，在病毒感染的MOI为0.01时，EC 50值分别为1.61μM，0.23μM和4.52μM，相应的治疗指数分别为15.22，696.96，20.56。在病毒感染的MOI为0.05时，EC 50值分别为1.95μM，0.62μM和3.63μM，相应的治疗指数分别为12.51，258.55，25.60。(见图2)
汉防己甲素、磷酸氯喹和瑞得西韦在病毒感染细胞后对SARS-Cov-2病毒具有抑制活性。
实施例3汉防己甲素对SARS-CoV-2的半数最大有效浓度(EC 50)
采取核酸定量法测定药物的EC 50。具体如下，提前一天将Vero细胞以约10000个/孔浓度接种至48孔板。用含有2％FBS(购自Gibco公司，货号16000044)的DMEM培养液将药物汉防己甲素配制成终浓度为20、10、5、2.5、1.25、0.625和0.3125μM，加入细胞中，放置37℃ 5％CO 2孵箱预处理1小时。然后，用含有2％FBS的DMEM培养液稀释SARS-CoV-2病毒(nCoV-2019BetaCoV/Wuhan/WIV04/2019分离株，GISAID登录号EPI_ISL_402124，由中国科学院武汉病毒研究所分离传代培养)，将 其加入到相应的孔中，使病毒载量为100TCID 50，37℃吸附培养2小时，弃病毒液，每孔加入不同浓度的汉方己甲素(200μl/孔)，使每孔中汉方己甲素的终浓度为20、10、5、2.5、1.25、0.625和0.3125μM，每个药物浓度设置3个复孔，并设立病毒对照和正常细胞对照组，放置37℃ 5％CO 2孵箱培养，每天观察细胞病变(CPE)。在感染后2天，每孔取50μl细胞上清提取核酸，用定量RT-PCR检测病毒载量。
RNA提取、RNA反转录和RT-PCR的方法均与实施例1相同。
依据病毒RNA拷贝数和CT值的换算公式：RNA Copies/mL＝CT*(-0.3)+13.17，计算病毒载量。采用公式(感染率(％)＝药物组RNA拷贝数/病毒对照组RNA拷贝数×100％)计算病毒在不同浓度药物处理下的感染率(％)，利用Graphpad Prism 7软件对数据进行S拟合分析，拟合结果如图3所示，并计算EC 50。
结果显示，汉防己甲素在细胞水平对SARS-CoV-2病毒具有抑制作用，其EC 50为8.99μM(图3)。
尽管本申请的具体实施方式已经得到详细的描述，本领域技术人员将会理解，根据已经公开的所有教导，可以对那些细节进行各种修改和替换，这些改变均在本申请的保护范围之内。本申请的全部范围由所附权利要求及其任何等同物给出。
参考文献：
Zaki AM,van Boheemen S,Bestebroer T M,et al.Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia[J].N Engl J Med,2012,367(19):1814-1820.
Zhong NS,Zheng BJ,Li YM,et al.Epidemiology and cause of severe acute respiratory syndrome(SARS)in Guangdong,People's Republic of China,in February,2003[J].The Lancet,2003,362(9393):1353-1358.
Cui J,Li F,Shi ZL.Origin and evolution of pathogenic coronaviruses[J].Nat Rev Microbiol,2019,17(3):181-192.
Wang M,Cao R,Zhang L,et al.Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus(2019-nCoV)in vitro[J].Cell Res,2020,0:1-3.

Claims (33)

  1. 通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，
    Figure PCTCN2020101581-appb-100001
    或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗冠状病毒引起的疾病或感染或冠状病毒引起的并伴有高血压的疾病或感染的产品中用途。
  2. 通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，
    Figure PCTCN2020101581-appb-100002
    或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗与冠状病毒有关且伴有高血压的肺部疾病或症状的产品中用途。
  3. 通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，
    Figure PCTCN2020101581-appb-100003
    或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于抑制冠状病毒在细胞(例如哺乳动物细胞)中复制或繁殖的产品中的用途，或在制备作为冠状病毒抑制剂的产品中的用途。
  4. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100004
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗与SARS-CoV-2有关的肺部疾病或症状，或无症状或有症状的SARS-CoV-2感染的产品中用途。
  5. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100005
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗与SARS-CoV-2有关且伴有高血压的肺部疾病或症状，或伴有高血压的无症状或有症状的SARS-CoV-2感染的产品中用途。
  6. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100006
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物在制备用于预防和/或治疗COVID-19或伴有高血压的COVID-19的产品中的用途，或在制备作为SARS-CoV-2抑制剂的产品中的用途，或在制备用于抑制SARS-CoV-2在细胞(例如哺乳动物细胞)中复制或繁殖的产品中的用途。
  7. 权利要求1-6任一项的用途，其中所述产品为人用药品或动物用兽药。
  8. 权利要求7的用途，其中所述产品为固体制剂、注射剂、喷剂、液体制剂、吸入制剂或复方制剂。
  9. 权利要求3或6的用途，其中所述哺乳动物包括牛科动物、马科动物、羊科动物、猪科动物、犬科动物、猫科动物、啮齿类动物、灵长类动物，例如是人，狗或猪。
  10. 权利要求1-6任一项的用途，其中所述的药物组合物还包含药学上可接受的载体或辅料，具体地，所述药物组合物为固体制剂、注射剂、吸入制剂、喷剂、液体制剂、或复方制剂。
  11. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100007
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于预防和/或治疗冠状病毒引起的疾病或感染。
  12. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100008
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于预防和/或治疗与冠状病毒有关且伴有高血压的肺部疾病或症状。
  13. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100009
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合 物的药物组合物，其用于抑制冠状病毒在细胞(例如哺乳动物细胞)中复制或繁殖。
  14. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100010
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其作为冠状病毒抑制剂。
  15. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100011
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于预防和/或治疗与SARS-CoV-2有关的肺部疾病或症状或无症状或有症状的SARS-CoV-2感染。
  16. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100012
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于预防和/或治疗与SARS-CoV-2有关且伴有高血压的肺部疾病或症状，或伴有高血压的无症状或有症状的SARS-CoV-2感染。
  17. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100013
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于预防和/或治疗COVID-19或伴有高血压的COVID-19。
  18. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100014
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其作为SARS-CoV-2抑制剂。
  19. 通式I所示的化合物，
    Figure PCTCN2020101581-appb-100015
    其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物，其用于抑制SARS-CoV-2在细胞(例如哺乳动物细胞)中复制或繁殖。
  20. 一种预防和/或治疗冠状病毒引起的疾病或感染或冠状病毒引起的并伴有高血压的疾病或感染的方法，其包括给有需要的宿主施用将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
  21. 一种预防和/或治疗与冠状病毒有关的肺部疾病或症状或与冠状病毒有关且伴有高血压的肺部疾病或症状的方法，其包括给有需要的宿主施用将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物的药物组合物。
  22. 一种有需要的宿主中抑制冠状病毒复制或繁殖的方法，其包括给有需要的宿主施用将预防和/或有效量的通式I所示的化合物，其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，或含有通式I所示的化合物，其几何异构体或其药学上可接 受的盐和/或其溶剂化物和/或其水合物的药物组合物。
  23. 权利要求20至22中任一项所述的方法，其中所述的冠状病毒为SARS-CoV-2。
  24. 权利要求20所述的方法，其中所述的冠状病毒引起的疾病为COVID-19。
  25. 权利要求20所述的方法，其中所述的冠状病毒引起的并伴有高血压的疾病为伴有高血压的COVID-19。
  26. 权利要求21所述的方法，其中所述的与冠状病毒有关的肺部疾病或症状为SARS-CoV-2有关的肺部疾病或症状。
  27. 权利要求21所述的方法，其中所述与冠状病毒有关且伴有高血压的肺部疾病或症状为SARS-CoV-2有关且伴有高血压的肺部疾病或症状。
  28. 权利要求20所述的方法，其中所述的冠状病毒引起的感染为无症状或有症状的SARS-CoV-2感染。
  29. 权利要求20所述的方法，其中所述的冠状病毒引起的并伴有高血压的感染为伴有高血压的无症状或有症状的SARS-CoV-2感染。
  30. 权利要求20至22中任一项所述的方法，其中所述宿主为哺乳动物。
  31. 权利要求20至22中任一项所述的方法，其中所述哺乳动物包括牛科动物、马科动物、羊科动物、猪科动物、犬科动物、猫科动物、啮齿类动物、灵长类动物，其中优选的哺乳动物是人，狗或猪。
  32. 权利要求13或19所述的化合物、其几何异构体或其药学上可接受的盐和/或其溶剂化物和/或其水合物，其中所述哺乳动物包括牛科动物、马科动物、羊科动物、猪科动物、犬科动物、猫科动物、啮齿类动物、灵长类动物，例如是人，狗或猪。
  33. 权利要求11-19任一项所述的药物组合物或权利要求20-22任一项所述的方法，其中所述的药物组合物还包含药学上可接受的载体或辅料，具体地，所述药物组合物为固体制剂、注射剂、吸入制剂、喷剂、液体制剂、或复方制剂。
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Non-Patent Citations (13)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "Tetrandrine Tablets Used in the Treatment of COVID-19 (TT-NPC)", CLINICALTRIALS.GOV, 16 March 2020 (2020-03-16), XP055842542, Retrieved from the Internet <URL:https://clinicaltrials.gov/ct2/show/NCT04308317> *
CUI JLI FSHI ZL: "Origin and evolution of pathogenic coronaviruses[J", NAT REV MICROBIOL, vol. 17, no. 3, 2019, pages 181 - 192, XP037065686, DOI: 10.1038/s41579-018-0118-9
DAN LI, JIANQIN JIANG: "Advances in Pharmacological Activities of Benzylisoquinoline Alkaloids", GUANGDONG CHEMICAL INDUSTRY, vol. 44, no. 9, 31 December 2017 (2017-12-31), pages 141 - 142, XP055842544 *
DONG KIM, JUNG MIN, MIN JANG, JUN LEE, YOUNG SHIN, CHUL PARK, JONG SONG, HYOUNG KIM, SEUNGTAEK KIM, YOUNG-HEE JIN, SUNOH KWON: "Natural Bis-Benzylisoquinoline Alkaloids-Tetrandrine, Fangchinoline, and Cepharanthine, Inhibit Human Coronavirus OC43 Infection of MRC-5 Human Lung Cells", BIOMOLECULES, vol. 9, no. 11, pages 696, XP055737811, DOI: 10.3390/biom9110696 *
JEON SANGEUN, KO MEEHYUN, LEE JIHYE, CHOI INHEE, BYUN SOO YOUNG, PARK SOONJU, SHUM DAVID, KIM SEUNGTAEK: "Identification of Antiviral Drug Candidates against SARS-CoV-2 from FDA-Approved Drugs", ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 64, no. 7, 23 June 2020 (2020-06-23), US, pages e00819 - 20, XP055825491, ISSN: 0066-4804, DOI: 10.1128/AAC.00819-20 *
LECHOWICZ KACPER, DROŻDŻAL SYLWESTER, MACHAJ FILIP, ROSIK JAKUB, SZOSTAK BARTOSZ, ZEGAN-BARAŃSKA MAŁGORZATA, BIERNAWSKA JOWITA, DA: "COVID-19: The Potential Treatment of Pulmonary Fibrosis Associated with SARS-CoV-2 Infection", JOURNAL OF CLINICAL MEDICINE, vol. 9, no. 6, 19 June 2020 (2020-06-19), XP055842538, DOI: 10.3390/jcm9061917 *
RUAN ZIJING, LIU CHAO, GUO YUTING, HE ZHENQING, HUANG XINHE, JIA XU, YANG TAI: "Potential Inhibitors Targeting RNA-Dependent RNA Polymerase Activity (NSP12) of SARS-CoV-2", 1 March 2020 (2020-03-01), pages 1 - 16, XP055803847, [retrieved on 20210511], DOI: 10.20944/preprints202003.0024.v1 *
See also references of EP4124337A4
SHEN LIANG, JUNWEI NIU, CHUNHUA WANG, BAOYING HUANG, WENLING WANG, NA ZHU, YAO DENG, HUIJUAN WANG, FEI YE, SHAN CEN, WENJIE TAN: "High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses", JOURNAL OF VIROLOGY, AMERICAN SOCIETY FOR MICROBIOLOGY, vol. 93, no. 12, 29 May 2019 (2019-05-29), XP055842533, DOI: 10.1128/JVI.00023-19 *
WANG MCAO RZHANG L ET AL.: "Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro[J", CELL RES, vol. 0, 2020, pages 1 - 3
XIUYUAN OU, LIU YAN, LEI XIAOBO, LI PEI, MI DAN, REN LILI, GUO LI, GUO RUIXUAN, CHEN TING, HU JIAXIN, XIANG ZICHUN, MU ZHIXIA, CHE: "Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV", NATURE COMMUNICATIONS, vol. 11, no. 1, XP055724997, DOI: 10.1038/s41467-020-15562-9 *
ZAKI AMVAN BOHEEMEN SBESTEBROER T M ET AL.: "Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia[J", N ENGL J MED, vol. 367, no. 19, 2012, pages 1814 - 1820, XP055274765, DOI: 10.1056/NEJMoa1211721
ZHONG NSZHENG BJLI YM ET AL.: "Epidemiology and cause of severe acute respiratory syndrome (SARS) in Guangdong, People's Republic of China, in February, 2003[J", THE LANCET, vol. 362, no. 9393, 2003, pages 1353 - 1358, XP004779115, DOI: 10.1016/S0140-6736(03)14630-2

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