HK40031254B - Use of substituted aminopropionate compounds in manufacture of a medicament for the treatment of sars-cov-2 infection - Google Patents

Description

Application of substituted amino propionate compound in preparing medicine for treating SARS-CoV-2 infection

Technical Field

The invention relates to a substituted amino propionic ester compound shown in the following formula I, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, and a pharmaceutical composition containing the compound, and application thereof in treating SARS-CoV-2 infection.

Background

Substituted aminopropionate compounds (compounds shown in formula I) have the chemical name of 2-ethylbutyl (2S) -2- [ [ (2R,3S,4R,5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazine-7-yl) -5-cyano-3, 4-dihydroxyoxacycloalkane-2-yl ] methoxyphenoxyphosphoryl ] amino ] propionate (2-ethylbutanyl (2S) -2- [ [ [ (2R,3S,4R,5R) -5- (4-aminopyrapryrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cya no-3, 4-dihydroxolan-2-yl ] methoxy-phenoxyphoryl ] amino ] propanoate), also known as Remdesivir (GS-5734), is a viral RNA polymerase inhibitor in its in vivo active form as the parent drug Nucleoside Triphosphate (NTP). The compound has already completed phase III clinical tests of clinical treatment of Ebola virus infection at present, and shows good treatment effect on the Ebola virus infection.

Remdesivir shows antiviral activity against EBOV and various variants of other filoviruses in cell-based assays. In the rhesus monkey model of ebola virus infection, even if treatment was initiated three days after virus exposure (systemic viral RNA was detected in two of six treated animals), intravenous administration of 10mg/kg of GS-5734 daily for 12 days significantly inhibited EBOV replication, protected 100% of EBOV infected animals from death, and improved clinical signs of disease and pathophysiological markers. In addition to ebola virus, GS-5734 also has broad-spectrum antiviral activity, such as Nipah virus (Nipah virus), middle east respiratory syndrome coronavirus (MERS-CoV), Marburg virus (Marburg virus), and the like.

2019 the novel coronavirus (2019-nCoV) is a new strain of coronavirus that has not previously been found in humans. On day 11/2/2020, the international committee for viral classification (ICTV) announced that the formal classification of the 2019 novel coronavirus (2019-nCoV) was named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). On the same day, the World Health Organization (WHO) announced that the formal name of the disease caused by this virus was COVID-19. The symptoms of SARS-CoV-2 virus infection are mainly pneumonia, and can be divided into simple infection, mild pneumonia, severe pneumonia, acute respiratory distress syndrome, sepsis, septic shock, etc. according to the severity of the disease. Patients with simple infections may have non-specific symptoms such as fever, cough, sore throat, nasal congestion, weakness, headache, muscle aches or discomfort, and the elderly and immunosuppressors may have atypical symptoms. Patients with mild pneumonia mainly have cough, dyspnea and tachypnea. Severe pneumonia can be seen in adolescents, adults or children, and the main symptoms are increased respiratory frequency, severe respiratory failure or dyspnea, cyanosis, lethargy, unconsciousness or convulsion, air suction and the like. The lung image of acute respiratory distress syndrome is a bilateral vitreography, but cannot be completely explained by effusion, lobular effusion or atelectasis or a lung mass image, and pulmonary edema is the main symptom. Septic patients often have fatal organ dysfunction, and septic shock is the most critical patient and has a high probability of death.

At present, aiming at SARS-CoV-2 virus infection, the support treatment is mainly used clinically, and no anti-virus medicine is available.

Disclosure of Invention

The invention aims to find a medicine with antiviral activity on SARS-CoV-2, which can be used for curing the related diseases caused by SARS-CoV-2 infection. The invention discovers that the compound of the formula I has the function of inhibiting the replication of SARS-CoV-2 through creative research, and has good potential treatment effect on the aspect of treating diseases caused by SARS-CoV-2.

The invention provides a compound shown as a formula I, a geometric isomer thereof, and a pharmaceutically acceptable salt and/or a solvate or hydrate thereof:

in certain embodiments, the pharmaceutically acceptable salts of the compounds of formula I described herein include inorganic or organic acid salts, as well as inorganic or organic base salts, and the invention relates to all forms of such salts including, but not limited to, sodium, potassium, calcium, lithium, meglumine, hydrochloride, hydrobromide, picrate, nitrate, sulfate, bisulfate, phosphate, biphosphate, acetate, propionate, butyrate, oxalate, pivalate, adipate, alginate, lactate, citrate, tartrate, succinate, maleate, fumarate, picrate, aspartate, gluconate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate, and the like.

The compound shown in the formula I can inhibit the replication of SARS-CoV-2 virus on cells and reduce the nucleic acid load of SARS-CoV-2 virus in cell culture.

After creative invention research, the inventors of the present invention found a new action characteristic of the compound shown in formula I in cells: the compound shown in the formula I can reduce the virus nucleic acid load level of SARS-CoV-2 infected cells under micromolar concentration.

The invention also relates to the use of the compound shown in the formula I, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in the preparation of medicaments for treating diseases or infections caused by SARS-CoV-2 (such as respiratory system diseases (such as simple infections, such as fever, cough, pharyngalgia and the like, pneumonia, acute respiratory tract infection, severe acute respiratory tract infection (SARI), hypoxic respiratory failure, acute respiratory distress syndrome, sepsis, septic shock and the like)),

the invention also relates to application of the compound shown in the formula I, geometric isomer thereof or pharmaceutically acceptable salt thereof and/or solvate thereof and/or hydrate thereof in preparing a medicament serving as a SARS-CoV-2 inhibitor.

The invention also relates to application of the compound shown in the formula I, geometric isomer thereof or pharmaceutically acceptable salt thereof and/or solvate thereof and/or hydrate thereof in preparing a medicament for inhibiting the replication or reproduction of SARS-CoV-2 in cells (such as mammalian cells).

The invention also relates to a pharmaceutical composition, which comprises the compound shown in the formula I, geometric isomers thereof or pharmaceutically acceptable salts thereof and/or solvates thereof and/or hydrates thereof,

preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant, and specifically, the pharmaceutical composition is a solid preparation, an injection, an external preparation, a spray, a liquid preparation, or a compound preparation.

The invention also relates to application of the pharmaceutical composition containing the compound shown in the formula I, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof or the compound shown in the formula I, the geometric isomer thereof and the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing medicines for treating respiratory diseases (such as simple infection, such as fever, cough, angina and the like, pneumonia, acute respiratory infection, Severe Acute Respiratory Infection (SARI), hypoxic respiratory failure, acute respiratory distress syndrome, sepsis, septic shock and the like) but not limited by respiratory diseases.

The invention also relates to application of the pharmaceutical composition containing the compound shown in the formula I, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing a medicament for treating diseases or infections caused by SARS-CoV-2 (such as respiratory system diseases (such as simple infections, such as fever, cough, pharyngalgia and the like, pneumonia, acute respiratory tract infection, severe acute respiratory tract infection (SARI), hypoxic respiratory failure, acute respiratory distress syndrome, sepsis, septic shock and the like)).

The invention also relates to the application of the pharmaceutical composition containing the compound of the formula I, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing the medicines serving as the SARS-CoV-2 inhibitor.

The invention also relates to the use of the pharmaceutical composition comprising a compound of formula I, a geometric isomer thereof, or a pharmaceutically acceptable salt thereof, and/or a solvate thereof and/or a hydrate thereof, for the preparation of a medicament for inhibiting the replication or propagation of SARS-CoV-2 in a cell (e.g., a mammalian cell).

The present invention also relates to a method for treating and/or preventing a disease in a mammal in need thereof or inhibiting SARS-CoV-2 replication or reproduction in a mammal in need thereof, which comprises administering to a mammal in need thereof a therapeutically and/or prophylactically effective amount of the pharmaceutical composition comprising the compound of formula I, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof or the compound of formula I, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, wherein the disease comprises a SARS-CoV-2 caused disease, for example, a viral infectious disease caused by SARS-CoV-2 (e.g., a respiratory disease, including simple infections such as fever, cough and sore throat, pneumonia, acute respiratory infection, Severe Acute Respiratory Infection (SARI), hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock, etc.).

The invention also relates to a compound shown in the formula I, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, which is used as a SARS-CoV-2 inhibitor.

The invention also relates to a compound shown in the formula I, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, which is used for inhibiting the replication or propagation of SARS-CoV-2 in cells (such as mammalian cells).

The invention also relates to a pharmaceutical composition containing the compound shown in the formula I, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, which is used as a SARS-CoV-2 inhibitor.

The invention also relates to a pharmaceutical composition containing the compound shown in the formula I, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof, which is used for inhibiting the replication or propagation of SARS-CoV-2 in cells (such as mammalian cells).

In certain embodiments, the SARS-CoV-2 caused disease described herein includes, but is not limited to, respiratory diseases, such as simple infections such as fever, cough, and angina, pneumonia, acute respiratory infections, Severe Acute Respiratory Infections (SARI), hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, and septic shock, among others.

In certain embodiments, the SARS-CoV-2 caused disease described herein is COVID-19.

In the present application, the formal classification of the term "2019 novel coronavirus (2019-nCoV)" is named Severe acid respiratory syndrome coronavirus 2 (SARS-CoV-2).

In the present application, the term "2019 novel coronavirus (2019-nCoV) -caused disease" is used under the formal name COVID-19.

In certain embodiments, the mammal comprises a bovine, equine, ovine, porcine, canine, feline, rodent, primate, wherein the preferred mammal is a human, cat, dog, or pig.

The pharmaceutical composition of the present invention can be prepared in various forms according to different administration routes.

According to the present invention, the pharmaceutical composition may be administered in any of the following ways: oral, aerosol inhalation, rectal, nasal, buccal, vaginal, topical, parenteral such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or via an external reservoir. Among them, oral, intraperitoneal or intravenous administration is preferable.

When administered orally, the compound of formula I, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof may be formulated into any orally acceptable formulation form, including but not limited to tablets, capsules, aqueous solutions or aqueous suspensions. Among them, the carriers generally used for tablets include lactose and corn starch, and additionally, a lubricant such as magnesium stearate may be added. Typical diluents used in capsule formulations include lactose and dried corn starch. Aqueous suspension formulations are generally prepared by mixing the active ingredient with suitable emulsifying and suspending agents. If desired, sweetening, flavoring or coloring agents may be added to the above oral dosage forms.

When administered rectally, the compounds of formula I, their geometric isomers or pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, are generally prepared in the form of suppositories by mixing the drug with a suitable non-irritating excipient. The excipient is in a solid state at room temperature and melts to release the drug at rectal temperature. Such excipients include cocoa butter, beeswax and polyethylene glycols.

When the compound shown in the formula I, the geometrical isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof can be prepared into different forms of topical preparations according to different affected surfaces or organs when the compound is applied topically, particularly for treating affected surfaces or organs which are easy to reach by topical application, such as eyes, skin or lower intestinal tract neurogenic diseases, and the specific description is as follows:

when topically administered to the eye, the compound of formula I, its geometric isomer, or a pharmaceutically acceptable salt thereof, and/or a solvate and/or hydrate thereof, can be formulated in the form of a micronized suspension or solution using sterile saline at a pH that is isotonic, with or without the addition of preservatives such as benzyl chloride alkoxides. In addition, for ophthalmic use, the compounds may also be formulated in the form of ointments such as vaseline.

When applied topically to the skin, the compounds of formula I, their geometric isomers or their pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, can be formulated into suitable ointment, lotion or cream formulations in which the active ingredient is suspended or dissolved in one or more carriers. The carrier that may be used in the ointment herein includes, but is not limited to: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; carriers that can be used in lotions or creams include, but are not limited to: mineral oil, sorbitan monostearate, tween 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

When topically administered to the lower intestinal tract, the compound of formula I, its geometric isomer, or a pharmaceutically acceptable salt thereof, and/or a solvate thereof and/or a hydrate thereof may be formulated in the form of a rectal suppository formulation or a suitable enema formulation as described above, and in addition, a topical transdermal patch may be used.

The compound shown in the formula I, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof can also be used for administration in the form of sterile injection preparations, including sterile injection water or oil suspensions, or sterile injection solutions. Among the carriers and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. In addition, the sterilized fixed oil may also be employed as a solvent or suspending medium, such as a monoglyceride or diglyceride.

The medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.

In the present invention, the term "therapeutically effective amount" or "prophylactically effective amount" refers to an amount sufficient to treat or prevent the disease in a patient, but low enough to avoid serious side effects (at a reasonable benefit/risk ratio), within the scope of sound medical judgment. The therapeutically effective amount of a compound will vary depending on the particular compound selected (e.g., taking into account the potency, effectiveness, and half-life of the compound), the route of administration selected, the disease being treated, the severity of the disease being treated, the age, size, weight, and physical condition of the patient being treated, the medical history of the patient being treated, the duration of the treatment, the nature of concurrent therapy, the desired therapeutic effect, and the like, but can nevertheless be routinely determined by one of ordinary skill in the art.

In addition, the specific dosage and application method of the compound of formula I, its geometric isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate for different patients depends on many factors, including the age, body weight, sex, natural health status, nutritional status, activity intensity of the drug, administration time, metabolic rate, severity of the disease and the subjective judgment of the treating physician. The preferred dosage is between 0.001 and 1000mg/kg body weight/day.

Drawings

FIG. 1 Remdesivir is effective in reducing the viral nucleic acid load on VeroE6 cells infected with SARS-CoV-2 virus. Wherein (a) Remdesivir is capable of inhibiting viral RNA load on cells 48h after the cells are infected with SARS-CoV-2 virus, wherein a drug concentration of 33 μ M reduces the nucleic acid load of the virus by one order of magnitude. The ordinate is the copy number of the viral RNA in the sample, and the abscissa is the drug concentration; (b) no cytotoxicity was observed with Remdesivir after 48h of treatment of test cells at the test concentrations. The ordinate is the percentage of cell viability relative to the negative control (cells only, no drug added) and the abscissa is the drug concentration.

Detailed Description

The following examples are illustrative of preferred embodiments of the present invention and are not to be construed as limiting the invention in any way.

Example 1: remdesivir reduction of viral nucleic acid load in SARS-CoV-2 virus infected cells

(1) Drug treatment of virus-infected cells

Vero E6 cells (purchased from ATCC under accession number 1586) were inoculated into 24-well plates and cultured for 24 h; then, virus infection was carried out, specifically, 2% cell maintenance solution (formulation: FBS (purchased from Gibco, Inc., Cat. No. 16000044) was added to MEM (purchased from Gibco, Cat. No. 10370021) in a volume ratio of 2%; namely, 2% cell maintenance solutionLiquid) SARS-CoV-2 (2019-nCoV) virus (nCoV-2019BetacoV/Wuhan/WIV04/2019 strain, supplied by Wuhan virus institute of Chinese academy of sciences) was diluted to the corresponding concentration, and then added to a 24-well plate to make each well contain 100TCID in terms of viral load₅₀. Next, Remdesivir (purchased from MedChemexpress, Cat. No. HY-104077) was diluted to the corresponding concentrations with 2% cell maintenance solution, added to the corresponding wells to give final concentrations of 100. mu.M, 33. mu.M, 11. mu.M, 3.7. mu.M, 1.23. mu.M, 0.41. mu.M, and 0.14. mu.M, and then subjected to 37 ℃ and 5% CO₂The incubator is continued for 48h, and only 2% cell maintenance solution without any test drug is added to the cell control group.

(2) RNA extraction

RNA extraction kits were purchased from Qiagen, cat # 74106. Consumables (spin columns, rnase-free 2ml collection tubes, etc.) and reagents (RLT, RW1, RPE, rnase-free water, etc.) involved in the RNA extraction steps described below were all components of the kit. The following extraction steps are all recommended by the kit instructions.

1) Taking 100 mu L of supernatant of a tested culture plate, adding the supernatant into a nuclease-free EP tube, then adding 350 mu L of Buffer RLT into each hole, uniformly blowing and sucking the mixture by using a pipette gun to ensure that the mixture is fully cracked, and centrifuging to take the supernatant;

2) adding equal volume of 70% ethanol into the supernatant obtained in the step 1), and uniformly mixing;

3) transferring the mixed solution obtained in the step 2) into a centrifugal column without RNA enzyme, centrifuging at 12000rpm for 15s, and discarding the waste liquid;

4) adding 700 mu L of Buffer RW1, centrifuging at 12000rpm for 15s, cleaning a centrifugal column, and discarding waste liquid;

5) adding 500 mu L Buffer RPE, centrifuging at 12000rpm for 15s, cleaning the centrifugal column, and discarding the waste liquid;

6) adding 500 μ L Buffer RPE, centrifuging at 12000rpm for 2min, cleaning the centrifugal column, and removing waste liquid;

7) replacing a new 2ml collecting tube without the RNase, centrifuging at 12000rpm for 1min, drying the centrifugal column, and then transferring the centrifugal column into the 1.5ml collecting tube in the step 8) integrally;

8) changing a new collection tube of 1.5ml, putting the centrifugation column dried in the step 7), adding 30. mu.l of RNase-free water into the centrifugation column, centrifuging at 12000rpm for 2min to obtain an eluate containing corresponding RNAs, adding RNase inhibitor (purchased from NEB company, product number M0314L), and detecting the concentration of each RNA by using Nano Drop (purchased from Thermo scientific, model number Nano Drop One).

(3) Reverse transcription of RNA

The reverse transcription of RNA was carried out using a reverse transcription Kit (PrimeScriptTMRT reagent Kit with gDNA Eraser, cat # RR047Q) manufactured by TaKaRa, as follows.

Removal of gDNA: RNA samples of each experimental group were collected, and 1. mu.g of each RNA sample was subjected to reverse transcription. Firstly, adding 2 μ l of 5 Xg DNA Eraser Buffer into RNA of each experimental group, complementing the reaction system to 10 μ l with RNase Free water, fully and uniformly mixing, and removing g DNA possibly existing in a sample in a water bath at 42 ℃ for 2 min;

reverse transcription: adding a proper amount of enzyme, a primer Mix and a reaction buffer solution into a sample obtained in the step one, complementing the volume to 20 mu l by RNase Free water, carrying out water bath reaction at 37 ℃ for 15min, and then putting into water at 85 ℃ for 5sec, so that cDNA can be obtained through transcription.

(4)Real-time PCR

And (3) detecting the copy number of the original virus solution contained in each milliliter by adopting fluorescent quantitative PCR.

The reaction system was mixed with TB Green Premix (Takara, Cat # RR820A) and amplification reaction and reading were performed on a StepOne Plus Real-time PCR instrument (brand: ABI). The number of copies per ml of the original virus solution was calculated. The method comprises the following steps:

firstly, establishing a standard product: the plasmid pMT-RBD (plasmid supplied by the institute of Wuhan Virus, national academy of sciences) was diluted to 5X 10⁸copies/μL,5×10⁷copies/μL,5×10⁶copies/μL, 5×10⁵copies/μL,5×10⁴copies/μL,5×10³copies/μL,5×10²copies/. mu.L. mu.L of the standard or cDNA template was used for the qPCR reaction.

② the primer sequences used in the experimental process are as follows (all shown in 5 '-3' direction):

RBD-qF:CAATGGTTTAACAGGCACAGG

RBD-qR:CTCAAGTGTCTGTGGATCACG

③ the reaction procedure is as follows:

pre-denaturation: 5 minutes at 95 ℃;

circulation parameters: 95 ℃ for 15 seconds, 54 ℃ for 15 seconds, 72 ℃ for 30 seconds, for a total of 40 cycles.

(5) Drug toxicity to cell test

The cytotoxicity of the drugs was measured using the CCK-8 kit (Beoytime). The method comprises the following specific steps:

inoculation of 1X 10 in 96-well plates⁴Vero E6(ATCC) cells were cultured at 37 ℃ for 8 hours.

② the drug was diluted with DMSO to an appropriate concentration of the stock solution, and then diluted with MEM medium (purchased from Gibco, cat. No. 10370021) containing 2% FBS (purchased from Gibco, cat. No. 16000044) to the same concentration as the drug treatment, the original medium in a 96-well plate was discarded, 100. mu.L of MEM medium containing the drug was added to the cells, and three replicate wells were made for each concentration. Negative controls (DMSO and media in well without drug) and blank controls (no cells, DMSO and media) were set. After the addition of the drug, the cells were cultured at 37 ℃ for 48 hours.

③ adding 20 mu L of CCK-8 solution (Beoytime) into the hole to be tested, mixing the solution gently without generating air bubbles, and culturing the solution for 2 hours at 37 ℃. OD readings were performed on a microplate reader (from Molecular Devices, model SpectraMax M5)₄₅₀And calculating the activity of the cells:

cell activity (%) ═ a_{(drug treatment group)}-A_{(blank control)})/(A_{(negative control)}-A_{(blank control)})×100％

Wherein A is the reading of the microplate reader.

(6) Results of the experiment

The results of the virus proliferation inhibition experiments showed that the test compounds were effective in inhibiting the replication of SARS-CoV-2 virus genome in the infection supernatant at concentrations of 100. mu.M, 33. mu.M, 11.1. mu.M and 3.7. mu.M (Table 1 and FIG. 1)

TABLE 1 in vitro antiviral assay of test Compounds (Remdesivir)

The cytotoxicity results show that treatment with the test compound (Remdesivir) did not alter cell viability at all concentrations tested, i.e. the test compound was not toxic to cells at all concentrations (table 2 and figure 1).

TABLE 2 cytotoxicity assay of test Compounds (Remdesivir)

SEQUENCE LISTING

<110> military medical research institute of military science institute of people's liberation force of China

Application of <120> substituted aminopropionic acid ester compound in treatment of SARS-CoV-2 infection

<130> IDC200202

<150> CN202010071087.7

<151> 2020-01-21

<160> 2

<170> PatentIn version 3.5

<210> 1

<211> 21

<212> DNA

<213> Artificial Sequence

<220>

<223> primer

<400> 1

caatggttta acaggcacag g 21

<210> 2

<211> 21

<212> DNA

<213> Artificial Sequence

<220>

<223> primer

<400> 2

ctcaagtgtc tgtggatcac g 21

Claims (23)

1. The application of the compound shown in the formula I or the pharmaceutically acceptable salt thereof in preparing the medicine for treating the diseases or the infections caused by SARS-CoV-2,

2. the use of claim 1, wherein the SARS-CoV-2 caused disease is a respiratory disease.

3. The use according to claim 1, wherein the SARS-CoV-2 caused disease is simple infection, pneumonia, acute respiratory infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis and/or septic shock.

4. The use of claim 3, wherein the acute respiratory infection is a severe acute respiratory infection.

5. The use of claim 3, wherein the simple infection is fever, cough, and/or sore throat.

6. Use of a pharmaceutical composition for the manufacture of a medicament for the treatment of a SARS-CoV-2 caused disease or infection, wherein the pharmaceutical composition comprises a compound of formula I or a pharmaceutically acceptable salt thereof,

7. the use of claim 6, wherein the SARS-CoV-2 caused disease is a respiratory disease.

8. The use according to claim 6, wherein the SARS-CoV-2 caused disease is simple infection, pneumonia, acute respiratory infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis and/or septic shock.

9. The use of claim 8, wherein the acute respiratory infection is a severe acute respiratory infection.

10. The use of claim 8, wherein the simple infection is fever, cough, and/or sore throat.

11. The use of claim 6, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant.

12. The use of claim 11, wherein the pharmaceutical composition is a solid formulation or a liquid formulation.

13. The use of claim 11, wherein the pharmaceutical composition is an injection or spray.

14. The use of claim 11, wherein the pharmaceutical composition is a combination formulation.

15. The use of claim 11, wherein the pharmaceutical composition is an external preparation.

16. The application of the compound shown in the formula I or the pharmaceutically acceptable salt thereof in preparing the medicine used as the SARS-CoV-2 inhibitor,

17. the application of the pharmaceutical composition in preparing the medicine used as SARS-CoV-2 inhibitor,

wherein the pharmaceutical composition comprises a compound of formula I or a pharmaceutically acceptable salt thereof.

18. The use of claim 17, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant.

19. The use of claim 18, wherein the pharmaceutical composition is a solid formulation or a liquid formulation.

20. The use of claim 18, wherein the pharmaceutical composition is an injection or spray.

21. The use of claim 18, wherein the pharmaceutical composition is a combination formulation.

22. The use of claim 18, wherein the pharmaceutical composition is an external preparation.

23. The use of any of claims 1-15, wherein the SARS-CoV-2 caused disease is COVID-19.