CN113952457A

CN113952457A - Application of termitimide or pharmaceutical composition containing termitimide in resisting coronavirus

Info

Publication number: CN113952457A
Application number: CN202010698299.8A
Authority: CN
Inventors: 肖飞; 王敏骏; 刘元; 仇波
Original assignee: Cinkate Pharmaceutical Intermediates Co ltd
Current assignee: Cinkate Pharmaceutical Intermediates Co ltd
Priority date: 2020-07-20
Filing date: 2020-07-20
Publication date: 2022-01-21

Abstract

The invention relates to application of tiramimide in resisting coronavirus. In particular, the present invention relates to teramitraz and pharmaceutical compositions thereof as (a) inhibitors of coronavirus; and/or (b) treating and/or preventing, relieving the related diseases caused by coronavirus infection. The tiramimide can treat and/or prevent and relieve related diseases such as respiratory tract infection, pneumonia (COVID-19) and the like caused by 2019 novel coronavirus infection.

Description

Application of termitimide or pharmaceutical composition containing termitimide in resisting coronavirus

Technical Field

The invention relates to the field of medicines, in particular to application of tiramimide or a pharmaceutical composition containing the same in resisting coronavirus.

Background

In acute infectious diseases, most of the infectious diseases are viral infectious diseases, the incidence rate of the viral infectious diseases is high, and the death rate is high. Because the detection and diagnosis means are limited, the outbreak of new epidemic caused by new viruses often has the characteristics of paroxysmal, random, unpredictable and the like, once the outbreak occurs, if no effective prevention and treatment means exists, the large-scale epidemic is very easy to cause, and the health and life safety of people is seriously threatened.

Coronaviruses (Coronaviruses) are single-stranded positive-strand RNA viruses belonging to the order Nidovirales (Nidovirales) Coronaviridae (Coronaviridae) orthocoronaviridae (orthocoronaviridae) and can infect various species such as humans, bats, pigs, mice, cows, horses, goats, monkeys, etc. There are 6 known human-infecting coronaviruses (HCoV), including the middle east respiratory syndrome-associated coronavirus (MERSR-CoV) and the severe acute respiratory syndrome-associated coronavirus (SARSr-CoV).

The novel coronavirus 2019-nCoV (SARS-CoV-2) is the 7 th coronavirus infecting human, and can cause severe pneumonia. The SARS-CoV-2 virus transmission pathway is not completely understood, and is known to be transmitted by droplets and contact, and there are human-borne and medical staff infections, certain community transmission risks, and the possibility of virus variation. There is currently no specific preventive or therapeutic approach for diseases caused by the novel coronavirus.

SARS-CoV-2 coronavirus belongs to the genus coronavirus of the family Coronaviridae, and is a single-stranded positive-sense RNA virus with an envelope. Similar to other known coronaviruses, the 2019-nCoV coronaviruses also complete the propagation of the offspring viruses through the processes of adsorption, penetration, uncoating, biosynthesis, assembly and release of the offspring viruses and the like. SARS-CoV-2 coronavirus infection of host cell begins with the binding of spike glycoprotein on the surface of virus envelope with receptor on the surface of host cell, and then membrane fusion occurs, the virus enters the host cell, under the action of cell organelles such as cell lysosome, the single-chain positive sense RNA of virus genetic material is released, and protein synthesis elements such as mitochondria and ribosome and essential elements of host cellThe polyprotein is produced by translation under the action of raw materials of (A) and the like, and then two essential cysteine proteases of 2019-nCoV coronavirus: papain (PL)^pro) And 3C-like protease (3C-like protease,3 CL)^pro) Cleavage at specific sites processes the polyprotein precursors, producing a number of non-structural proteins that are important to the viral life cycle. Under the action of these nonstructural proteins, the viral RNA replicates the progeny viral nucleic acid material and translates into the desired structural proteins in large quantities, completing assembly and release of the progeny virus. Any link or key enzyme in the life cycle of SARS-CoV-2 coronavirus infected cell can be used as research target of antiviral medicine, such as cysteine protease PL for hydrolytic cleavage of polyprotein precursor^proAnd 3CL^proRNA polymerase responsible for completing replication of progeny viral genetic material, and the like.

At present, no specific vaccine or antiviral drug exists for severe pneumonia diseases caused by SARS-CoV-2 coronavirus. These infectious diseases seriously affect the life health of people, and the development of antiviral drugs with good effect is imminent. Aiming at SARS-CoV-2 coronavirus, a low-toxicity and high-efficiency antiviral drug is developed to meet the clinical requirements of SARS-CoV-2 coronavirus infected patients at home and abroad, and has great social significance.

In view of the above, there is an urgent need in the art to develop inhibitors against SARS-CoV-2 coronavirus for the treatment of pneumonia caused by infection with the novel coronavirus.

Disclosure of Invention

The invention aims to provide an active ingredient capable of effectively inhibiting virus replication and a new application thereof in related diseases caused by virus infection.

In particular, the invention provides the use of pharmaceutical compositions containing tiramistat against coronaviruses, in particular against novel coronaviruses (SARS-CoV-2).

In a first aspect of the invention, there is provided the use of a pharmaceutical composition comprising:

(a1) a first active ingredient which is tiramimide or a pharmaceutically acceptable salt thereof or a crystalline hydrate thereof or a solvate thereof or a prodrug thereof:

(a2) a second active ingredient selected from the group consisting of: remdesivir (Ruidesacvir or GS-5734), Favipiravir (favipiravir), Galidesivir, GS-441524, NHC (EIDD-1931), EIDD-2801, GC-376, Lopinavir (Lopinavir), Ritonavir (Ritonavir), Nelfinavir (Nelfinavir); chloroquine (Chloroquin), hydroxychloroquine (hydroxychloroquine), cyclosporin (cyclosporine), clarithromycin (Carrimycin), baicalin (baicain), baicalein (baicalein), phillyrin (forsythoside), chlorogenic acid (chlorogenic acid), emodin (emodin), mycophenolic acid (mycophenolic acid), Mycophenolate mofetil (Mycophenolate mofetil), Naphthoquine (Naphthoquin), Ciclesonide (Ciclesonide), Ribavirin (Ribavirin), Penciclovir (Penticlovir), Leflunomide (Leflunomide), Teriflunomide (Teriflumonide), nafamostat (nafamostat), nitrozonide (nitrozanixonide), Darunavir (Darunavir), Abiravir (Salicovir), Salicornib (Ribovir), Ribovir (Ribovir), Ribovinifer (Ribovir), Ribovir (Ribovir), Ribovinil), Ribovir (Ribovir), Ribovir (Ribovinil), Ribovir (Ribovir), Ribovinil), Ribovir (Ribovir), Ribovir (Ribovinil), Ribovir (Ribovir), Ribovinil), Ribovir (Ribovinil), Ribovir (Ribovir), Ribovir (Ribovinil), Ribovir (Ribovir), Ribovinil), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir (Ribovir), Ribovir

MEDI-557, A-60444(RSV-604), MDT-637, BMS-433771, or a pharmaceutically acceptable salt thereof, or a combination thereof;

and (b) a pharmaceutically acceptable carrier.

And, the pharmaceutical composition is used for preparing (a) an inhibitor for inhibiting replication of coronavirus; and/or (b) a medicament for the treatment and/or prevention, amelioration of a related disease caused by a coronavirus infection.

In another preferred embodiment, the first active ingredient is sodium teriumide.

In another preferred embodiment, the second active ingredient is selected from: reed-civir, leflunomide, or a combination thereof.

In another preferred embodiment, the pharmaceutical composition is used for the preparation of (a) an inhibitor that inhibits 2019 novel coronavirus replication; and/or (b) a medicament for treating and/or preventing, ameliorating, or preventing a related disease caused by the 2019 novel coronavirus infection.

In another preferred embodiment, the coronavirus is selected from the group consisting of: (1) human infecting coronavirus: SARS-CoV (Severe acute respiratory syndrome coronavirus), 2019 novel coronavirus (2019-nCoV or SARS-CoV-2), MERS-CoV (Middle East respiratory syndrome coronavirus)

(2) Common cold causing coronavirus: the common cold-causing coronavirus is preferably selected from the group consisting of: human coronavirus OC43(Human coronavirus OC43), Human coronavirus229E (Human coronavirus229E), Human coronavirus NL63(Human coronavirus NL63), Human coronavirus HKUl (Human coronavirus HKUl);

in another preferred embodiment, the coronavirus-induced related disease is selected from the group consisting of:

(D1) common cold, high risk symptom infection, respiratory tract infection, pneumonia and complications thereof caused by human coronavirus infection;

(D2) SARS-CoV-2-induced novel coronavirus pneumonia (Corona Virus Disease 2019, COVID-19);

(D3) any combination of the above.

In another preferred embodiment, the administration mode of the pharmaceutical composition comprises: oral and non-oral.

In another preferred embodiment, the pharmaceutical composition comprises: powder, granule, capsule, injection, tincture, oral liquid, tablet, buccal tablet, or dripping pill.

In a second aspect of the present invention, there is provided a use of an active ingredient which is tiramimide or a pharmaceutically acceptable salt thereof or a crystalline hydrate thereof or a solvate thereof or a prodrug thereof, or a formulation containing the active ingredient;

and, said formulation is used for the preparation of (a) an inhibitor for inhibiting coronavirus replication; and/or (b) a medicament for the treatment and/or prevention, amelioration of a related disease caused by a coronavirus infection.

In another preferred embodiment, the formulation is used to prepare (a) an inhibitor that inhibits 2019 novel coronavirus replication; and/or (b) a medicament for treating and/or preventing, ameliorating, or preventing a related disease caused by the 2019 novel coronavirus infection.

In another preferred embodiment, the formulation comprises: oral and non-oral formulations.

In another preferred embodiment, the formulation comprises: powder, granule, capsule, injection, tincture, oral liquid, tablet, buccal tablet, or dripping pill.

In a third aspect of the invention, there is provided a method of inhibiting viral replication comprising the steps of:

contacting a first pharmaceutical composition with the coronavirus, thereby inhibiting replication of the coronavirus;

wherein the pharmaceutical composition comprises: (a1) a first active ingredient which is tiramimide or a pharmaceutically acceptable salt thereof or a crystalline hydrate thereof or a solvate thereof or a prodrug thereof; and/or

(a2) A second active ingredient, preferably Remdesivir (Reed-West or GS-5734), Leflunomide (Leflunomide).

In another preferred embodiment, the second active ingredient is selected from: remdesivir (Ruidesacvir or GS-5734), Favipiravir (favipiravir), Galidesivir, GS-441524, NHC (EIDD-1931), EIDD-2801, GC-376, Lopinavir (Lopinavir), Ritonavir (Ritonavir), Nelfinavir (Nelfinavir); chloroquine (Chloroquine), hydroxychloroquine (hydroxychloroquine), cyclosporine (cyclosporine), clarithromycin (Carrimycin), baicalin (baicain), baicalein (baicalein), forsythoside (forsythoside), chlorogenic acid (chlorogenic acid), emodin (emodin), mycophenolic acid (mycophenolic acid), Mycophenolate mofetil (Mycophenolate mofetil), Naphthoquine (Naphthoquine), Ciclesonide (Ciclesonide), Ribavirin (Ribavirin), Penciclovir (Penticlovir), Leflunomide (Leflunomide), Teriflunomide (Terifluoride), nafamostat (nafamostat), nitrozonide (nitroxanide),Darunavir (Darunavir), Abidol (Arbidol), Camostat (Camostat), Niclosamide (Niclosamide), Baritinib (Baricitinib), Ruxolitinib (Ruxolitinib), Dasatinib (Dasatinib), Saquinavir (Saquinavir), Beclaabuvir, Semetivir (Simeprevir), palivizumab, Movizumab (Motavizumab), RSV-IGIV

in another preferred embodiment, the method is an in vitro method and is non-therapeutic and non-diagnostic.

In a fourth aspect of the present invention, there is provided a method of treating, preventing, and/or ameliorating a related disease caused by a 2019 novel coronavirus, comprising the steps of: administering to a subject in need thereof a safe and effective amount of a first pharmaceutical composition; wherein the first pharmaceutical composition is as described above.

Drawings

FIG. 1 is a graph showing the inhibition rate of sodium teriumide against SARS-CoV-2, and its cytotoxicity.

FIG. 2 is a bar graph of inhibition of SARS-CoV-2 replication with Teramit sodium alone and in combination with Retzivir.

Figure 3 is a bar graph of cytotoxicity of toremifide sodium alone and in combination with resisitev.

Detailed Description

The inventor of the present invention has conducted extensive and intensive studies and, through a large number of screenings, has unexpectedly developed for the first time the use of a pharmaceutical composition comprising tiramite for combating coronaviruses. The present invention has been completed based on this finding.

Specifically, the invention discloses a pharmaceutical composition containing teriflunomide, redciclovir, leflunomide or a combination thereof, which can effectively inhibit the replication and activity of SARS-CoV-2 virus, and therefore can be used for preparing an inhibitor for inhibiting 2019 novel coronavirus replication; and/or preventing, relieving and treating related diseases caused by SARS-CoV-2. Has good clinical application prospect.

Term(s) for

As used herein, "an active compound of the present invention", "an active ingredient of the present invention", "tiramimide of the present invention", "an active compound of the present invention inhibiting coronavirus replication" are used interchangeably and refer to tiramimide having an excellent activity of inhibiting coronavirus replication, including tiramimide, or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a prodrug thereof, or a combination thereof.

As used herein, "formulation of the invention" refers to a formulation containing an active compound of the invention.

As used herein, the term "comprising" or variations thereof, such as "comprises" or "comprising," etc., are understood to imply the inclusion of stated elements or components but not the exclusion of any other elements or components.

As used herein, the terms "novel coronavirus", "2019-nCov" or "SARS-CoV-2" are used interchangeably, the 2019 novel coronavirus being the 7 th coronavirus known to infect humans and causing new coronary pneumonia (COVID-19), one of the serious infectious diseases threatening global human health.

Coronavirus (coronavirus)

Coronaviruses (CoV) belong to the family of the Nidovirales (Nidovirales) Coronaviridae (Coronaviridae), a enveloped positive-strand RNA virus, a subfamily of which contains four genera, alpha, beta, delta and gamma.

Among the coronaviruses currently known to infect humans, HCoV-229E and HCoV-NL63 belong to the genus alpha coronavirus, and HCoV-OC43, SARS-CoV, HCoV-HKU1, MERS-CoV and SARS-CoV-2 are all the genus beta coronavirus. SARS-CoV-2 is also known as 2019-nCoV.

Highly pathogenic coronaviruses SARS-CoV and MERS-CoV, which outbreak in 2003 and 2012, respectively, both belong to the genus beta coronavirus. The novel coronavirus (SARS-CoV-2) which is outbreak in 2019 and has 80% similarity with SARS-CoV and 40% similarity with MERS-CoV, and also belongs to the beta genus coronavirus.

The genome of the virus is a single-strand positive-strand RNA, is one of RNA viruses with the largest genome, and codes comprise replicase, spike protein, envelope protein, nucleocapsid protein and the like. In the initial stage of viral replication, the genome is translated into two peptide chains of up to several thousand amino acids, the precursor Polyprotein (Polyprotein), which is subsequently cleaved by proteases to yield nonstructural proteins (e.g., RNA polymerase and helicase) and structural proteins (e.g., spike protein) and accessory proteins.

Active Compounds and active ingredients of the invention

In the present invention, a pharmaceutical composition effective for inhibiting replication of coronaviruses, particularly 2019 novel coronaviruses (SARS-CoV-2) is provided.

In the present invention, the pharmaceutical composition comprises: (a1) a first active ingredient which is tiramimide or a pharmaceutically acceptable salt thereof or a crystalline hydrate thereof or a solvate thereof or a prodrug thereof:

(a2) a second active ingredient selected from the group consisting of: remdesivir (Ruidesacvir or GS-5734), Favipiravir (favipiravir), Galidesivir, GS-441524, NHC (EIDD-1931), EIDD-2801, GC-376, Lopinavir (Lopinavir), Ritonavir (Ritonavir), Nelfinavir (Nelfinavir); chloroquine (Chloroquine), hydroxychloroquine (hydroxychloroquine), cyclosporin (cyclosporine), clarithromycin (Carrimycin), baicalin (baicain), baicalein (baicalein), phillyrin (forsythoside), chlorogenic acid (chlorogenic acid), emodin (emodin), mycophenolic acid (mycophenolic acid), Mycophenolate mofetil (Mycophenolate mofetil), Naphthoquine (Naphthoquine), Ciclesonide (Ciclesonide), Ribavirin (Ribavirin), Penciclovir (Penticlovir), Leflunomide (Leflunomide), Teriflunomide (Teriflumonide), nafamostat (nafamostat), nitrozonide (nitronide), Darunavir (Darunavir), abacavir (dabir), salicylic acid (Ribovir), quinavir (Ribovir), Ribovir (Ribovir), Ribovinil (Ribovir), Ribovir (Ribovinil), Ribovir (Ribovir), Ribovinil), Ribovir (Ribovir), Ribovir (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinil (Ribovinil), Ribovinabuvir, Simeprevir, palivizumab, Motavizumab, RSV-IGIV

and (b) a pharmaceutically acceptable carrier.

Experiments show that the pharmaceutical composition can effectively inhibit 2019 novel coronavirus (SARS-CoV-2) replication, thereby preventing, treating and/or relieving SARS-CoV-2 related diseases.

As used herein, "the first active compound of the present invention", "the active compound of the present invention inhibiting viral replication" are used interchangeably and refer to a compound having an excellent activity of inhibiting viral replication, including terremimide or a pharmaceutically acceptable salt thereof, or a crystal thereof, or a solvate thereof. The terms "second active compound", "second active ingredient" and "second active ingredient" are used interchangeably herein and refer to other antiviral drugs that are preferably either reed-solomon or leflunomide.

The chemical formulas of a part of the first and second active ingredients of the present invention are shown in the following table 1:

table 1 chemical structural formula of the first active ingredient or the second active ingredient:

it is to be understood that the first active ingredient of the present invention comprises tiramimide, or a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof, or a prodrug thereof. It is to be understood that the active ingredients of the present invention also include crystalline, amorphous, and deuterated forms of the active compounds of the present invention.

The "pharmaceutically acceptable salts" are conventional non-toxic salts formed by the reaction of the active compounds of the present invention with inorganic or organic acids. For example, conventional non-toxic salts can be prepared by reacting the active compounds of the present invention with inorganic acids including hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, sulfamic acid, phosphoric acid and the like, or organic acids including citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, isethionic acid and the like; or sodium, zinc, potassium, calcium, aluminum or ammonium salts of the active compounds of the invention formed with propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, aspartic acid or glutamic acid as esters and then with inorganic bases; or the corresponding inorganic acid salt formed by the active compound of the invention and lysine, arginine and ornithine and then hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid or phosphoric acid, or the corresponding organic acid salt formed by the active compound of the invention and formic acid, acetic acid, picric acid, methanesulfonic acid or ethanesulfonic acid; or 1-4 phosphate groups in the molecule of the active compound of the present invention form salts with 1-4 molecules of organic amines, including but not limited to trimethylamine monophosphate, triethylamine monophosphate, tri-n-butylamine monophosphate, trimethylamine triphosphate, triethylamine triphosphate, tri-n-butylamine triphosphate. .

In addition, the active ingredients of the invention are particularly suitable for use in combination with other antiviral or anti-coronavirus drugs. Representative other antiviral or anti-coronavirus agents include (but are not limited to): interferons, RNA-dependent RNA polymerase inhibitors (e.g., Remdesivir (Ruidevir or GS-5734), Favipiravir (favipiravir), Galidesivir, GS-441524, NHC, EIDD-2801); 3CL protease inhibitors (e.g. protease inhibitorsGC-376), Lopinavir (Lopinavir), Ritonavir (Ritonavir), Nelfinavir (Nelfinavir); chloroquine (Chloroquin, Sigma-C6628), hydroxychloroquine, cyclosporin (cyclosporine), clarithromycin (Carrimycin), baicalin (baicain), baicalein (baicalein), phillyrin (forsythoside), chlorogenic acid (chlorogenic acid), emodin (emodin), mycophenolic acid (mycophenolic acid), Mycophenolate mofetil (Mycophenolate mofetil), Naphthoquine (Naphthoquinone), Ciclesonide (Ciclesonide), Ribavirin (Ribavirin), Penciclovir (Penciclovir), Leflunomide (Leflunomide), Teriflunomide (Teriflumominide), nafamostat (nafamostat), nitrozonamide (Nitazoxanide), Darunavir (Darunavir), Arbidol (Arbidol), Sativib (Sapridinil), Salacitinib (Sapridinavir), Nipavinib (Savinifer), Savinifer (Ribax), Ribavirucin (Rivasil), Rivastigrinil (Rivasil), Rivasil (Rivasil), Rivastigrinil (Rivasicilin), Rivastigrinil (Rivasil), Rivastigrinil (Rivasil), Rivasil (Rivastigrinil), Rivasil (Rivasil), Rivasil (R (Rivasil), Rivasil (Rivasil), Rivascil (Rivastigrinil), Rivasil (Rivascil (Rivasil), Rivasil (R (Rivasil), Rivascil (Rivasil), Rivasil (Rivasil), Rivascil (Rivasil (R (Rivasil), Rivascil (R (Rivasil), Rivasil (R (Rivasil), Rivasil (Rivasil), Rivasil (Rivascil (Rivasil), Rivasil (R (Rivasil), Rivasil (Rivasil), Rivasil (R (Rivasil), Rivasil (R (Rivasil), Rivasil (R (Rivasil), Rivasil (R (Rivasil), Rivastati (Rivasil (Rivastati (R (Rivasil), Rivastati (R (Rivastati (R), Rivasil (R), Rivasil), Ri

MEDI-557, A-60444(RSV-604), MDT-637, BMS-433771, or a pharmaceutically acceptable salt thereof, or a combination thereof. The interferon comprises one or more of interferon alpha-2 a, interferon alpha-2 b, interferon alpha-n 1, interferon alpha-n 3, interferon beta-1 a and interferon beta-1 b.

Additional active therapeutic agents for the treatment of respiratory symptoms and sequelae of infection may also be used in combination with tiramimide. The other agents are preferably administered orally or by direct inhalation. For example, other preferred additional therapeutic agents for use in combination with teremide in the treatment of respiratory tract infections include, but are not limited to, bronchodilators and corticosteroids.

Glucocorticoids, originally introduced as asthma therapy in 1950 (Carryer, Journal of Allergy,21, 282-287,1950), remained the most effective and consistently effective therapy for this disease, but their mechanism of action was not fully understood (Morris, J.allergy Clin. Immunol.,75(1Pt)1-13,1985). Unfortunately, oral glucocorticoid therapy is associated with profound undesirable side effects such as central obesity, hypertension, glaucoma, glucose intolerance, accelerated cataract formation, bone mineral loss, and psychological effects, all of which limit their use as long-term therapeutics (Goodman and Gilman, 10 th edition, 2001). The solution to systemic side effects is the delivery of steroid drugs directly to the site of inflammation. Inhaled Corticosteroids (ICS) have been developed to alleviate the serious side effects of oral steroids. Non-limiting examples of corticosteroids that may be used in combination with tiramimide are dexamethasone, dexamethasone sodium phosphate, fluoromethalone acetate, loteprednol etabonate, hydrocortisone, prednisolone, fludrocortisone, triamcinolone acetonide, betamethasone, beclomethasone dipropionate, methylprednisolone, fluocinolone acetonide, flunisolide, fluocorten-21-butyl ester (fluocortin-21-butylate), flumethasone pivalate, budesonide, halobetasol propionate, mometasone furoate, fluticasone propionate, ciclesonide; or a pharmaceutically acceptable salt thereof.

Other anti-inflammatory agents that act through an anti-inflammatory cascade mechanism may also be used as additional therapeutic agents for the treatment of viral respiratory infections in combination with tiramite. The use of "anti-inflammatory signal transduction modulators" (referred to herein as AISTM) such as phosphodiesterase inhibitors (e.g. PDE-4, PDE-5 or PDE-7 specific), transcription factor inhibitors (e.g. blocking NF κ B by IKK inhibition) or kinase inhibitors (e.g. blocking P38MAP, JNK, PI3K, EGFR or Syk) is a logical approach to cut inflammation as these small molecules target a limited number of common intracellular pathways-those signal transduction pathways that are key points for anti-inflammatory therapeutic intervention (see reviewed by p.j. barnes, 2006). These non-limiting additional therapeutic agents include: 5- (2, 4-difluoro-phenoxy) -1-isobutyl-1H-indazole-6-carboxylic acid (2-dimethylamino-ethyl) -amide (P38Map kinase inhibitor ARRY-797); 3-cyclopropylmethoxy-N- (3, 5-dichloro-pyridin-4-yl) -4-difluoromethoxy-benzamide (PDE-4 inhibitor roflumilast); 4- [2- (3-cyclopentyloxy-4-methoxyphenyl) -2-phenyl-ethyl ] -pyridine (PDE-4 inhibitor CDP-840); n- (3, 5-dichloro-4-pyridinyl) -4- (difluoromethoxy) -8- [ (methylsulfonyl) amino ] -1-dibenzofurancarboxamide (omister, a PDE-4 inhibitor); n- (3, 5-dichloro-pyridin-4-yl) -2- [1- (4-fluorobenzyl) -5-hydroxy-1H-indol-3-yl ] -2-oxo-acetamide (PDE-4 inhibitor AWD 12-281); 8-methoxy-2-trifluoromethyl-quinoline-5-carboxylic acid (3, 5-dichloro-1-oxy-pyridin-4-yl) -amide (PDE-4 inhibitor Sch 351591); 4- [5- (4-fluorophenyl) -2- (4-methanesulfinyl-phenyl) -1H-imidazol-4-yl ] -pyridine (P38 inhibitor SB-203850); 4- [4- (4-fluoro-phenyl) -1- (3-phenyl-propyl) -5-pyridin-4-yl-1H-imidazol-2-yl ] -but-3-yn-1-ol (P38 inhibitor RWJ-67657); 4-cyano-4- (3-cyclopentyloxy-4-methoxy-phenyl) -cyclohexanecarboxylic acid 2-diethylamino-ethyl ester (2-diethyl-ethyl ester prodrug of cilomilast, PDE-4 inhibitor); (3-chloro-4-fluorophenyl) - [ 7-methoxy-6- (3-morpholin-4-yl-propoxy) -quinazolin-4-yl ] -amine (gefitinib, EGFR inhibitor); and 4- (4-methyl-piperazin-1-ylmethyl) -N- [ 4-methyl-3- (4-pyridin-3-yl-pyrimidin-2-ylamino) -phenyl ] -benzamide (imatinib, EGFR inhibitor).

Combinations that include inhaled beta 2-adrenoceptor agonist bronchodilators such as formoterol, salbutamol or salmeterol and teromit are also suitable but non-limiting combinations that may be used to treat viral infections of the respiratory tract.

Inhaled β 2-adrenoceptor agonist bronchodilators such as formoterol or salmeterol in combination with ICS are also used to treat both bronchoconstriction and inflammation. Combinations comprising these combinations of ICS and β 2-adrenoceptor agonists, together with tiramimide, are also suitable, but non-limiting combinations useful for the treatment of viral infections of the respiratory tract.

Anticholinergic agents have potential uses for the treatment or prevention of pulmonary bronchoconstriction and are therefore useful as additional therapeutic agents for the treatment of viral respiratory infections in combination with tiramite. These anticholinergics include, but are not limited to, muscarinic receptor (particularly the M3 subtype) antagonists that have shown therapeutic efficacy in humans for the control of cholinergic properties in COPD (Witek, 1999); 1- { 4-hydroxy-1- [3,3, 3-tris- (4-fluoro-phenyl) -propionyl ] -pyrrolidine-2-carbonyl } -pyrrolidine-2-carboxylic acid (1-methyl-piperidin-4-ylmethyl) -amide; 3- [3- (2-diethylamino-acetoxy) -2-phenyl-propionyloxy ] -8-isopropyl-8-methyl-8-azonia (azonia) -bicyclo [3.2.1] octane (ipratropium-N, N-diethylglycinate); 1-cyclohexyl-3, 4-dihydro-1H-isoquinoline-2-carboxylic acid 1-aza-bicyclo [2.2.2] oct-3-yl ester (solifenacin); 2-hydroxymethyl-4-methanesulfinyl-2-phenyl-butyric acid 1-aza-bicyclo [2.2.2] oct-3-yl ester (revatonyl); 2- {1- [2- (2, 3-dihydro-benzofuran-5-yl) -ethyl ] -pyrrolidin-3-yl } -2, 2-diphenyl-acetamide (darifenacin); 4-azepan-1-yl-2, 2-diphenyl-butanamide (mebendazole); 7- [3- (2-diethylamino-acetoxy) -2-phenyl-propionyloxy ] -9-ethyl-9-methyl-3-oxa-9-azonian-tricyclo [3.3.1.02,4] nonane (oxytropin-N, N-diethylglycinate); 7- [2- (2-diethylamino-acetoxy) -2, 2-di-thiophen-2-yl-acetoxy ] -9, 9-dimethyl-3-oxa-9-azonian-tricyclo [3.3.1.02,4] nonane (tiotropium bromide-N, N-diethylglycinate); dimethylamino-acetic acid 2- (3-diisopropylamino-1-phenyl-propyl) -4-methyl-phenyl ester (tolterodine-N, N-dimethylglycine ester or salt); 3- [4, 4-bis- (4-fluoro-phenyl) -2-oxo-imidazolin-1-yl ] -1-methyl-1- (2-oxo-2-pyridin-2-yl-ethyl) -pyrrolidinium; 1- [1- (3-fluoro-benzyl) -piperidin-4-yl ] -4, 4-bis- (4-fluoro-phenyl) -imidazolidin-2-one; 1-cyclooctyl-3- (3-methoxy-1-aza-bicyclo [2.2.2] oct-3-yl) -1-phenyl-prop-2-yn-1-ol; 3- [2- (2-diethylamino-acetoxy) -2, 2-di-thiophen-2-yl-acetoxy ] -1- (3-phenoxy-propyl) -1-azoniabicyclo [2.2.2] octane (aclidinium bromide-N, N-diethylglycine ester or salt); or (2-diethylamino-acetoxy) -di-thiophen-2-yl-acetic acid 1-methyl-1- (2-phenoxy-ethyl) -piperidin-4-yl ester.

In addition, since SARS-CoV-2 infection can cause acute lung injury, the active ingredient of the present invention is particularly suitable for use in combination with a drug having an effect of ameliorating acute lung injury. Representative drugs include, but are not limited to, Zinc (Zinc), Fingolimod (Fingolimod), vitamin c (vitamin c), Olmesartan Medoxomil (olmestanan Medoxomil), valsartan (valsartan), Losartan (Losartan), Thalidomide (Thalidomide), glycyrrhizic acid (glycyrrhetic acid), Artemisinin (Artemisinin), dihydroartemisinin (dihydroartemisinin), Artesunate (Artesunate), Artemisone (artemisinine), Azithromycin (Azithromycin), Escin (Escin), Naproxen (Naproxen). The Zinc (Zinc) includes but is not limited to Zinc sulfate, Zinc glycyrrhetate, and Zinc gluconate.

The active ingredient of the invention can inhibit the infection activity of novel coronavirus such as SARS-CoV-2. Therefore, when the active ingredient of the present invention is therapeutically administered or administered, the infection with the novel coronavirus of 2019 (SARS-CoV-2) can be inhibited, thereby achieving an antiviral effect.

Tiramimide may also be combined with mucolytics to treat both infection and respiratory infection symptoms. A non-limiting example of a mucolytic is ambroxol. Likewise, teremide can be combined with an expectorant to treat both infection and respiratory infection symptoms. A non-limiting example of an expectorant is guaifenesin.

Nebulized hypertonic saline is used to improve the immediate and long-term clearance of small airways in patients with pulmonary diseases (Kuzik, j. pediatrics2007, 266). Tiramite may also be combined with nebulized hypertonic saline, particularly when the paramyxoviridae virus infects bronchiolitis. The combination of tiramistat and hypertonic saline may also include any of the additional agents discussed above. In a preferred aspect, about 3% nebulized hypertonic saline is used.

It is also possible to combine any of the compounds of the invention with one or more additional active therapeutic agents in a unit dosage form for simultaneous or sequential administration to a patient. The combination therapy may be administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more administrations.

Co-administration of a compound of the invention with one or more additional active therapeutic agents generally refers to the simultaneous or sequential administration of a compound of the invention and one or more additional active therapeutic agents such that a therapeutically effective amount of both the compound of the invention and the one or more additional active therapeutic agents are present in the patient.

Co-administration includes administering a unit dose of a compound of the invention before or after administering a unit dose of one or more additional active therapeutic agents, e.g., within seconds, minutes, or hours of administering one or more additional active therapeutic agents. For example, a unit dose of a compound of the invention may be administered first, followed by a unit dose of one or more additional active therapeutic agents within seconds or minutes. Alternatively, a unit dose of one or more other therapeutic agents may be administered first, followed by a unit dose of a compound of the invention within seconds or minutes. In some instances, it may be desirable to first administer a unit dose of a compound of the invention, and after a period of hours (e.g., 1-12 hours), a unit dose of one or more additional active therapeutic agents. In other cases, it may be desirable to first administer a unit dose of one or more additional active therapeutic agents, followed by a unit dose of a compound of the invention after a period of hours (e.g., 1-12 hours).

Combination therapy can provide "synergy" and "synergy", i.e., the effect obtained when the active ingredients are used together is greater than the sum of the effects obtained when the compounds are used separately. When the active ingredients are: (1) are co-formulated and administered or delivered simultaneously in a combined formulation; (2) alternatively as separate formulations or delivered in parallel; or (3) delivered by some other administration regimen, a synergistic effect may be obtained. When delivered in alternating treatments, synergy can be achieved when the compounds are administered or delivered sequentially, e.g., in separate tablets, pills or capsules, or by different injections with separate syringes. Typically, during alternating treatments, an effective dose of each active ingredient is administered sequentially, i.e. consecutively, whereas in combination treatments, an effective dose of two or more active ingredients are administered together. Synergistic antiviral effects mean antiviral effects greater than the predicted net additive effects of the individual compounds in the combination.

In yet another embodiment, a method of inhibiting viral RNA polymerase in a cell is provided, comprising contacting a cell infected with a virus with an effective amount of tiramit, or a pharmaceutically acceptable salt, solvate, and/or prodrug (e.g., ester) thereof, thereby inhibiting viral RNA polymerase.

In yet another embodiment, a method of inhibiting viral RNA polymerase in a cell is provided, comprising contacting a cell infected with a virus with an effective amount of tiramit, or a pharmaceutically acceptable salt, solvate, and/or prodrug (e.g., ester) thereof, and at least one additional active therapeutic agent, thereby inhibiting viral RNA polymerase.

In yet another embodiment, a method of inhibiting viral RNA polymerase in a cell is provided, comprising contacting a cell infected with a virus with an effective amount of tiramimide, or a pharmaceutically acceptable salt, solvate, and/or prodrug (e.g., ester) thereof, and at least one additional active therapeutic agent of choice.

In yet another embodiment, a method of treating a respiratory viral infection in a human or other mammal is provided, comprising administering to the human or other mammal a therapeutically effective amount of tiramistat, or a pharmaceutically acceptable salt, solvate, and/or prodrug (e.g., ester) thereof.

In yet another embodiment, there is provided a method of treating a respiratory viral infection in a human or other mammal comprising administering to the human or other mammal a therapeutically effective amount of tiramistat, or a pharmaceutically acceptable salt, solvate, and/or prodrug (e.g., ester) thereof, and at least one additional active therapeutic agent, thereby inhibiting respiratory viral RNA polymerase.

In yet another embodiment, a method of treating a respiratory viral infection in a human or other mammal is provided, comprising administering to the human or other mammal a therapeutically effective amount of tiramimide, or a pharmaceutically acceptable salt, solvate, and/or prodrug (e.g., ester) thereof, and at least one additional active therapeutic agent.

Pharmaceutical composition and application

The invention also provides application of the active compound for inhibiting virus replication, or one or more mixtures of pharmaceutically acceptable salts or prodrugs thereof as an active ingredient in preparing medicaments for treating and/or preventing and relieving respiratory tract infection, pneumonia and other related diseases caused by infection of viruses (especially 2019 novel coronavirus) such as coronavirus, influenza virus, respiratory syncytial virus and/or Porcine Epidemic Diarrhea Virus (PEDV).

The pharmaceutical composition provided by the present invention preferably contains 0.001-99 wt% of active ingredient, preferably 0.1-90 wt% or 1-50 wt% of active compound of the present invention as active ingredient, the rest being pharmaceutically acceptable carrier, diluent or solution or salt solution.

If necessary, one or more pharmaceutically acceptable carriers can be added into the medicine. The carrier comprises diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants and the like which are conventional in the pharmaceutical field.

The compounds and pharmaceutical compositions provided herein may be in a variety of forms such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, and the like, and may be presented in suitable solid or liquid carriers or diluents and in suitable sterile devices for injection or instillation.

Various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional preparation methods in the pharmaceutical field. The unit dose of the formulation generally comprises 0.05 to 1000mg of active compound according to the invention, preferably 1 to 500mg of active compound according to the invention.

The compounds and pharmaceutical compositions of the present invention may be administered to mammals in the clinical setting, including humans and animals, by oral, nasal, dermal, pulmonary or gastrointestinal routes of administration. Most preferably oral. Most preferably, the daily dose is 0.01-400mg/kg body weight, and is administered once or in portions of 0.01-200mg/kg body weight. Regardless of the method of administration, the optimal dosage for an individual will depend on the particular treatment. Usually starting with a small dose and gradually increasing the dose until the most suitable dose is found.

The drug or inhibitor of the present invention can be administered by a variety of different means, e.g., by injection, spray, nasal drop, eye drop, osmotic, absorption, physical or chemical mediated methods, into the body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue; or mixed with other materials or encapsulated and introduced into body.

Typically, the active ingredient of the invention or a pharmaceutical composition containing it is administered in unit dosage form, either enterally or parenterally, such as orally, intravenously, intramuscularly, subcutaneously, nasally, oromucosally, ocularly, pulmonary and respiratory, dermally, vaginally, rectally, and the like.

The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including O/W type, W/O type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion and liniment; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.

The active ingredients of the invention can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle delivery systems.

In order to formulate the active ingredients of the present invention into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agent can be water, ethanol, isopropanol, etc.; the adhesive can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, and the like.

The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.

To prepare the administration units into capsules, the active ingredients of the invention can be mixed with diluents, glidants and the mixture placed directly into hard or soft capsules. Or mixing the effective components with diluent, binder, and disintegrating agent, making into granule or pellet, and placing into hard capsule or soft capsule. The diluents, binders, wetting agents, disintegrants, glidants used to prepare the tablets of the invention may also be used to prepare the capsules of the invention.

In order to prepare the active ingredients of the invention into injection, water, ethanol, isopropanol, propylene glycol or the mixture of the water, the ethanol, the isopropanol and the propylene glycol can be used as a solvent, and a proper amount of solubilizer, cosolvent, pH regulator and osmotic pressure regulator which are commonly used in the field can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, mannitol and glucose can be added as proppant for preparing lyophilized powder for injection.

In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired.

The active ingredient or composition of the present invention can be administered alone or in combination with other therapeutic or symptomatic agents.

When the active ingredient of the present invention acts synergistically with other therapeutic agents, its dosage should be adjusted according to the actual circumstances.

The main advantages of the invention include:

(a) the active compound of the invention can effectively inhibit SARS-CoV-2 replication of the viruses EC₅₀The value is small, and can reach the level of 27.07 mu M.

(b) The active compounds according to the invention have low toxic side effects (cytotoxicity CC)₅₀271.6 mu M), the medicine property is good. This suggests that the tiramimide of the invention has good medicinal prospect in the field of resisting new coronary pneumonia.

(c) The active compound of the invention is combined with the Reidesvir, and can more effectively inhibit the replication of SARS-CoV-2 and other viruses under the condition of low dosage level (20 mu M +1 mu M).

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally followed by conventional conditions, such as Sambrook et al, molecular cloning: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989), or according to the manufacturer's recommendations. Unless otherwise indicated, percentages and parts are by weight.

Experimental viral strains and cell strains

The SARS-CoV-2 virus used for detection is a clinical isolate which is separated and preserved by Wuhan virus institute of Chinese academy of sciences. Viral infection experiments were all performed in the BSL-3 laboratory.

The cell line used for detection is monkey kidney cell line Vero E6, and is subcultured in the laboratory. Cell culture conditions: culturing in DMEM medium supplemented with 10% fetal calf serum, 100U/mL penicillin and 100. mu.g/mL streptomycin at 37 ℃ and 5% CO₂Culturing in an incubator.

Experimental reagent and instrument

The powder of the tested sample CK8 (sodium tallowmitate, purity 94.1%, batch number: A2170303R, production date: 2017.03.16) is provided by pharmaceutical chemical intermediate (Shanghai) of Xinkai. CK8 powder 16mg plus DMSO 485. mu.L was dissolved in stock solution of 100mM concentration, dispensed and stored at-20 ℃. The control drug was Remdesivir (MCE Corp.).

The reagents and consumables used in the experiment mainly comprise DMEM medium (Gibco), Fetal Bovine Serum (FBS), pancreatin, PBS buffer solution, CellTiter Glo cell viability testA Kit, a Viral Nucleic Acid Extraction Kit (Viral Nucleic Acid Extraction Kit II, Geneaid), a novel coronavirus Nucleic Acid fluorescence PCR detection Kit (United medicine), and a 96-well cell culture plate. The instrument used for the experiment mainly comprises CO₂Incubator, biological safety cabinet, fluorescent quantitative PCR instrument (BioRad), Synergy LX multifunctional microplate detector (BioTek).

Test example 1: cytotoxicity experiments:

after 24h of cell culture, 100. mu.L of culture supernatant was gently aspirated, 50. mu.L of CellTiter-Glo reagent was added, and the cells were incubated at room temperature for 10min with 3 shaking cycles. And then transferring all cell lysates into a totally black 96-well plate, and detecting Lum luminescent signals on a Synergy LX multifunctional microplate detector. And calculating the toxicity of the compound to the cells according to the luminescence value.

The experimental results are as follows: the results of the cytotoxicity experiments are shown in table 2.

TABLE 2 Telomide sodium, and its cytotoxicity in combination with Reidesvir

Test example 2: study of inhibitory Effect of Compounds on replication of novel Coronaviridae

After the cells are cultured for 24h, 100 mu L of culture supernatant is taken from each hole, 400 mu L of VB Lysis buffer solution in the virus nucleic acid extraction kit is added, the incubation is carried out for 10min at room temperature, and the virus is inactivated and lysed. Viral RNA was extracted according to the kit instructions. Then, a reaction system was prepared according to the instructions of the novel coronavirus nucleic acid detection kit (Table 3). The fluorescent quantitative PCR reaction was set up in the fluorescent quantitative PCR instrument according to the procedure of Table 4 to detect ORF1ab gene of the virus.

Table 3: RT-PCR reaction system

RT-PCR reaction solution	19μL
		PCR MIX	1μL
RNA	5μL

Table 4: RT-PCR reaction procedure

The antiviral activity and cytotoxicity of the compound are represented by the inhibition rate (%) and cytotoxicity (%) of the compound against viral replication. The calculation formula is as follows:

cytotoxicity (%) 100-sample reading/cell control mean 100 [% ]

Inhibition (%) 100-

And (3) data analysis: EC of the Compound₅₀Value sum CC₅₀Values were calculated for fit using GraphPad Prism software.

Mode (2): log (inhibitor) vs. normalized response- -Variable slope

The experimental results are as follows: as shown in table 5:

TABLE 5 Teramit sodium, and its use in combination with Redsivir for the inhibition of replication of the novel coronavirus (SARS-CoV-2)

In addition, by combining the data of the example 1 and the example 2, the cytotoxicity effect of the tested sample is obtained by testing the activity of the Vero E6 cells after the drug treatment; the replication level of Vero E6 cells infected by the virus after the drug treatment is detected by a qRT-PCR method to obtain the virus replication of a detected sampleInhibiting effect. All data are fitted by GraphPad Prism software to calculate inhibition rate and cytotoxicity curve, and EC of the compound is obtained₅₀Value sum CC₅₀The results are shown in FIG. 1.

As can be seen from FIG. 1, the EC of sodium teriumide for inhibiting SARS-Co-2 replication₅₀The value was 27.07. mu.M, CC₅₀The value is 271.6 mu M, and the sodium teriummate can inhibit the replication of SARS-CoV-2 under the condition of extremely small concentration, and the generated cytotoxicity is lower. Only when the concentration is close to 300 μm, significant toxicity to cells is caused. Therefore, the compound has good clinical medicinal prospect.

FIG. 2 is a bar graph of inhibition of SARS-CoV-2 replication in Redsivir alone and in combination with TAIROMETER sodium. As can be seen from FIG. 2, when the concentration of Reidsevir was the same, the addition of the combination of Teramit sodium also increased the replication inhibition of SARS-CoV-2, and the combination was more effective in inhibiting the replication of SARS-CoV-2.

Figure 3 is a bar graph of cytotoxicity of rescixivir alone and in combination with termitimide sodium. As can be seen from fig. 3, when the concentration of the ridciclovir is 1, 2 and 4 μm, the cytotoxicity is increased along with the increase of the administration concentration, but the influence on the cells is not large, and the total dose is not more than 30%; when the concentration of the Reineckr is the same, the cytotoxicity is not obviously increased and is not more than 30 percent in total when the Tairemomide sodium is added for combined use.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. Use of a pharmaceutical composition comprising:

and (b) a pharmaceutically acceptable carrier.

2. The use according to claim 1, wherein the pharmaceutical composition is used for the preparation of (a) an inhibitor that inhibits 2019 novel coronavirus replication; and/or (b) a medicament for treating and/or preventing, ameliorating, or preventing a related disease caused by the 2019 novel coronavirus infection.

3. The use according to claim 1, wherein the coronavirus is selected from the group consisting of: (1) human infecting coronavirus: SARS-CoV (Severe acute respiratory syndrome coronavirus), 2019 novel coronavirus (2019-nCoV or SARS-CoV-2), MERS-CoV (Middle East respiratory syndrome coronavirus)

(2) Common cold causing coronavirus: the common cold-causing coronavirus is preferably selected from the group consisting of: human coronavirus OC43(Human coronavirus OC43), Human coronavirus229E (Human coronavirus229E), Human coronavirus NL63(Human coronavirus NL63), and Human coronavirus HKUl (Human coronavirus HKUL).

4. The use of claim 1, wherein the disease associated with coronavirus is selected from the group consisting of:

(D3) any combination of the above.

5. The use of claim 1, wherein the pharmaceutical composition is administered by a method comprising: oral and non-oral.

6. Use of an active ingredient or a formulation containing the active ingredient, wherein the active ingredient is tiramimide or a pharmaceutically acceptable salt thereof or a crystalline hydrate thereof or a solvate thereof or a prodrug thereof;

7. The use of claim 6, wherein the formulation is used to prepare (a) an inhibitor that inhibits 2019 novel coronavirus replication; and/or (b) a medicament for treating and/or preventing, ameliorating, or preventing a related disease caused by the 2019 novel coronavirus infection.

8. The use of claim 6, wherein said formulation comprises: oral and non-oral formulations.

9. A method of inhibiting viral replication comprising the steps of:

10. A method of treating, preventing, and/or ameliorating a related disorder caused by a 2019 novel coronavirus, comprising the steps of: administering to a subject in need thereof a safe and effective amount of a first pharmaceutical composition; wherein the first pharmaceutical composition is as described above.