Application of betrixaban or pharmaceutically acceptable salt thereof

Abstract

The invention discloses an application of betrixaban or a pharmaceutically acceptable salt thereof. Drug screening and in-vitro drug experiments find that betrixaban or pharmaceutically acceptable salts thereof can inhibit virus replication and infection ability, and can block virus-induced syncytial bodies, so that cell death caused by the syncytial bodies is reduced. In addition, the betrixaban or the pharmaceutically acceptable salt thereof provided by the invention has broad-spectrum antiviral activity, and also has the characteristics of safety and high efficiency.

Description

[0001] Cross-reference to related applications This application is a divisional application filed on April 15, 2025, with application number CN202510468474.7, entitled “Use of betrixaban or a pharmaceutically acceptable salt thereof in the preparation of an antiviral drug”, which is incorporated herein by reference in its entirety. Technical Field

[0002] This invention belongs to the field of biomedicine, and in particular relates to the use of betrixaban or a pharmaceutically acceptable salt thereof in the preparation of antiviral drugs. Background Technology

[0003] In recent years, due to changes in the natural environment and the expansion of human activities, viral diseases have occurred frequently, posing a significant threat to human public health and safety. The spread of viral diseases is a major public health and safety issue threatening human health. Therefore, there is an urgent need to develop a safe, broad-spectrum, and highly effective antiviral drug. Summary of the Invention

[0004] To address at least some of the technical problems in the prior art described above, the present invention provides the use of betroxaban or a pharmaceutically acceptable salt thereof. Specifically, the present invention includes the following.

[0005] A first aspect of the invention provides the use of betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative or metabolite thereof in the preparation of an antiviral medicament.

[0006] In some embodiments, according to the application described in the present invention, the virus comprises a DNA virus and / or an RNA virus.

[0007] In some embodiments, according to the application described in the present invention, the DNA virus includes at least one of herpes simplex virus type 1, herpes simplex virus type 2, varicella-zoster virus, cytomegalovirus, smallpox virus, monkeypox virus, adenovirus, human papillomavirus, parvovirus, bocavirus, and hepatitis B virus.

[0008] In some embodiments, according to the application described in the present invention, the RNA virus includes at least one of vesicular stomatitis virus, encephalomyocarditis virus, mouse hepatitis virus, influenza A virus, influenza B virus, influenza C virus, dengue virus, Zika virus, Ebola virus, Marburg virus, Nipah virus, coronavirus, hepatitis A virus, hepatitis C virus, rotavirus, measles virus, human immunodeficiency virus, respiratory syncytial virus, and rabies virus.

[0009] In some embodiments, the use according to the present application, wherein the anti-viral comprises at least one of the following: (1) inhibiting viral entry into a host cell; (2) inhibiting viral genome replication; (3) inhibiting viral protein processing or assembly; (4) inhibiting viral release; (5) reducing the amount or activity of a virus.

[0010] In some embodiments, the use according to the present application, wherein the anti-viral is achieved by administering to the subject a therapeutically effective amount of the drug.

[0011] In some embodiments, the use according to the present application, wherein the subject comprises a mammal.

[0012] In some embodiments, the use according to the present application, wherein the mammal comprises a human.

[0013] In some embodiments, the use according to the present application, wherein the therapeutically effective amount is 0.01-1000 mg / Kg.

[0014] In a second aspect, the present application provides a use of betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative or metabolite thereof in combination with other drugs in the preparation of a combined anti-viral drug.

[0015] The present application discloses that betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative or metabolite thereof can inhibit viral replication and infection ability, and can block syncytia induced by viruses, thereby reducing cell death caused by syncytia. In addition, betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative or metabolite thereof has broad-spectrum anti-viral activity, and has the characteristics of safety and high efficiency. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 Inhibition of vesicular stomatitis virus (VSV) by betrixaban in human fibrosarcoma cells (2fTGH cells) is shown.

[0017] Figure 2 Inhibition of viral transcription and syncytia formation by betrixaban is shown.

[0018] Figure 3 Inhibition of VSV by betrixaban in other cells is shown.

[0019] Figure 4 Results of studies showing the antiviral mechanism of betrixaban are shown. DETAILED DESCRIPTION

[0020] Various exemplary embodiments of the present application are now described in detail, it being understood that the foregoing description is not intended to be limiting of the application. Rather, the description is provided as illustrative of certain aspects, features and embodiments of the present application.

[0021] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. In addition, where particular ranges of values are given, it is to be understood that each intervening value, as well as each value now known or to be discovered, between the ranges is included. The various embodiments of the present application are now described in detail.

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described. All patents, patent applications, publications, and descriptions mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the patents, patent applications, publications, and descriptions are cited.

[0023] Application In one aspect of the present application, there is provided a use of betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative or metabolite thereof in the manufacture of an antiviral medicament.

[0024] The term "pharmaceutically acceptable salt" as used herein refers to organic and inorganic salts of the compound of the present application, betrixaban. Pharmaceutically acceptable salts are well known in the art, examples of which include, but are not limited to: inorganic acid salts, such as hydrochloride, hydrobromide, phosphate, sulphate, perchlorate; organic acid salts, such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate; or salts obtained by other methods known in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, ascorbate, aspartate, benzenesulphonate, benzoate, bisulphate, borate, butyrate, camphorate, camphorsulphonate, cyclopentanepropionate, digluconate, dodecylsulphate, ethanesulphonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulphate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulphonate, lactobionate, lactate, laurate, laurylsulphate, malate, malonate, mesylate, 2-naphthalenesulphonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulphate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulphonate, undecanoate, valerate and the like.

[0025] The term "solvate" as used herein refers to an association or complex of one or more solvent molecules and a compound of the present application. Solvents include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol and the like.

[0026] The term "hydrate" as used herein refers to an association or complex of solvent molecules, which are water, and a compound of the present application.

[0027] The term "isomer" as used herein includes enantiomeric, diastereomeric, and geometric (or conformational) isomers of a given structure. For example, the present application includes compounds which are single stereoisomers, enantiomeric mixtures, diastereomeric mixtures, and geometric (or conformational) isomer mixtures. Unless otherwise specified, the present application includes all tautomeric forms of the structures disclosed herein for betrixaban.

[0028] The term "prodrug" as used herein represents a compound which in vivo is converted to a compound of the following formula (I) or formula (II): Formula (I); Formula (II).

[0029] The term "prodrug" refers to a pharmacologically inactive derivative of a parent drug molecule that requires a biotransformation, either spontaneous or enzymatic, in the body to release the active drug. Prodrugs are variants or derivatives of the compounds of the present application that have groups cleavable under metabolic conditions. When prodrugs undergo solvolysis or enzymatic degradation under physiological conditions, they become the compounds of the present application that are pharmaceutically active in vivo. Prodrug forms often provide the advantages of solubility, tissue compatibility, or delayed release in mammalian organisms (see Bundgard, Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985 and Silverman, The Organic Chemistry of Drug Design and Drug Action, pp. 352-401, Academic Press, San Diego, CA, 1992). Prodrugs known in the art include acid derivatives well known to those skilled in the art, for example, esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with an amine, or a basic group with a base to form an acylated base derivative.

[0030] The term "metabolite" as used herein refers to a product produced through metabolism of a specified compound or salt thereof in the body. Metabolic products of a compound can be identified using techniques known in the art, and their activities can be determined using methods known in the art. Such products can be produced, e.g., by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, and the like. Accordingly, the present application includes metabolites of compounds of Formula (I) or (II), including those produced by contact of a compound of Formula (I) or (II) with a mammal or a cell line derived therefrom for a period of time sufficient to yield the metabolites.

[0031] In the present application, the term "derivative" refers to a product derived from an atom or functional group in a compound of Formula (I) or (II) by substitution with another atom or functional group, which is not specifically limited, including but not limited to esterification, acylation, or metal complexation of a compound of Formula (I) or (II), and the like.

[0032] In the present application, the virus includes DNA virus and / or RNA virus. Examples of the DNA virus include, but are not limited to, herpes simplex virus type 1, herpes simplex virus type 2, varicella-zoster virus, cytomegalovirus, smallpox virus, monkeypox virus, adenovirus, human papillomavirus, parvovirus, bocavirus, hepatitis B virus, etc. Examples of the RNA virus include, but are not limited to, vesicular stomatitis virus, encephalomyocarditis virus, mouse hepatitis virus, influenza A virus, influenza B virus, influenza C virus, dengue virus, Zika virus, Ebola virus, Marburg virus, Nipah virus, coronavirus, hepatitis A virus, hepatitis C virus, rotavirus, measles virus, human immunodeficiency virus, respiratory syncytial virus, rabies virus, etc. In a preferred embodiment, the DNA virus is herpes simplex virus type 1. In another preferred embodiment, the RNA virus is vesicular stomatitis virus. In yet another preferred embodiment, the RNA virus is encephalomyocarditis virus. In yet another preferred embodiment, the RNA virus is mouse hepatitis virus. In yet another preferred embodiment, the RNA virus is influenza A virus.

[0033] In the present application, "antiviral" refers to ameliorating a condition, either prior to or following the onset of the disease or disorder. The extent of such amelioration or prevention can be at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100% as measured by any standard in the art, as compared to a reference group under the same conditions but without treatment. Beneficial or desired clinical results include, but are not limited to, either detectable or undetectable results including alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether partial or complete. Antiviral situations include, but are not limited to, (1) inhibition of viral entry into a host cell; (2) inhibition of viral genome replication; (3) inhibition of viral protein processing or assembly; (4) inhibition of viral release; (5) reduction in the amount or activity of the virus.

[0034] In the present application, antiviral is achieved by administering to a subject a therapeutically effective amount of the drug. The subject includes, but is not limited to, a mammal. Examples of the mammal include, but are not limited to, a human, a mouse, a rabbit, a cat, a dog, a cow, a sheep, a pig, etc.

[0035] In the present application, the medicament contains a therapeutically effective amount of betrixaban or its pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative or metabolite, particularly betrixaban maleate, and optionally, a pharmaceutically acceptable excipient or carrier. The term "pharmaceutically acceptable excipient or carrier" as used herein refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the agents from one organ, or portion of the body, to another organ, or portion of the body. Each excipient or carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

[0036] In the present application, the administration of the medicament is not particularly limited, and representative administration modes include, but are not limited to, oral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and topical administration. Accordingly, the medicament of the present application can be prepared into various dosage forms acceptable in clinic, including oral dosage forms, injection dosage forms, topical administration dosage forms or external use dosage forms, etc.

[0037] The therapeutically effective amount according to the present application means a pharmaceutically effective administration dose, i.e. the amount of active compound (i.e. betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analogue, derivative or metabolite thereof, in particular betrixaban maleate) sufficient to significantly improve the condition without causing serious side effects. The daily administration dose of betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analogue, derivative or metabolite thereof is usually 0.01-1000 mg / Kg, preferably 0.01-500 mg / Kg, or 0.01-400 mg / Kg, or 0.01-300 mg / Kg, or 0.01-200 mg / Kg, or 0.01-150 mg / Kg, or 0.01-100 mg / Kg, or 0.01-50 mg / Kg, or 0.01-40 mg / Kg, or 0.01-30 mg / Kg, most preferably 0.01-20 mg / Kg. Exemplary effective administration doses are, for example, 0.01 mg / Kg, 0.05 mg / Kg, 0.1 mg / Kg, 0.2 mg / Kg, 0.3 mg / Kg, 0.4 mg / Kg, 0.5 mg / Kg, 0.75 mg / Kg, 0.95 mg / Kg, 1 mg / Kg, 1.25 mg / Kg, 1.5 mg / Kg, 1.75 mg / Kg, 2 mg / Kg, 2.5 mg / Kg, 2.75 mg / Kg, 3 mg / Kg, 3.25 mg / Kg, 3.5 mg / Kg, 3.75 mg / Kg, 4 mg / Kg, 4.25 mg / Kg, 4.5 mg / Kg, 4.75 mg / Kg, 5 mg / Kg, 5.25 mg / Kg, 5.5 mg / Kg, 5.75 mg / Kg, 6 mg / Kg, 6.25 mg / Kg, 6.5 mg / Kg, 6.75 mg / Kg, 7 mg / Kg, 7.25 mg / Kg, 7.5 mg / Kg, 7.75 mg / Kg, 8 mg / Kg, 8.25 mg / Kg, 8.5 mg / Kg, 8.75 mg / Kg, 9 mg / Kg, 9.25 mg / Kg, 9.5 mg / Kg, 9.75 mg / Kg, 10 mg / Kg, 11 mg / Kg, 12 mg / Kg, 13 mg / Kg, 14 mg / Kg, 15 mg / Kg, 16 mg / Kg, 17 mg / Kg, 18 mg / Kg, 19 mg / Kg, 20 mg / Kg. Preferably, the above daily administration dose is based on betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analogue, derivative or metabolite thereof. The single dose can be administered once daily, can be administered in several portions per day, or can be used intermittently.

[0038] Combined application One aspect of the invention provides the use of betrixaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative or metabolite thereof with other drugs in the preparation of a combination antiviral drug.

[0039] In this invention, other drugs include, but are not limited to, remdesivir, ribavirin, favipiravir, acyclovir, valacyclovir, famciclovir, ganciclovir, brivudine, interferon, oseltamivir phosphate, peramivir, zanamivir, mabaloxavir, favipiravir, arbidol, etc.

[0040] Example The following example illustrates the inhibitory effect of betrixaban on the virus.

[0041] 1. Inhibitory effect of betrixaban on VSV virus in 2fTGH cells 1.1 Experimental Methods 2fTGH cells were treated with betrixaban (in DMSO) and VSV at concentrations of 10, 40, 60, and 80 μM, respectively. DMSO+VSV served as the control group. At 12 h and 24 h after treatment, the expression levels of VSV gene and protein were detected by qPCR and Western Blot, respectively. At the same time, the viral particle content in the cell supernatant was detected by plaque assay.

[0042] 1.2 Experimental Results The results are as follows Figure 1 As shown, specifically, qPCR results indicated that with increasing betraxaban concentration, the relative expression level of VSV viral genes decreased, and the inhibitory effect of the drug on viral transcription was not enhanced by the duration of drug action. Figure 1 (A). Western blot results showed that, relative to the internal control protein, the expression level of VSV viral protein decreased with increasing betrixaban concentration. Figure 1 B), Figure 1 The concentrations of betrixaban in group B, from left to right, were 20, 40, and 80 μM, respectively. Viral plaque assay results showed that the viral titer decreased significantly with increasing betrixaban concentration. Compared to the DMSO group, the viral load was almost zero when using 60 μM betrixaban, indicating a negative correlation between viral replication and infectivity and betrixaban concentration. Figure 1 (C).

[0043] In summary, betrixaban can reduce VSV viral transcription and expression, and inhibit viral replication and infectivity.

[0044] 2. Inhibitory effects of betrixaban on other viruses 2.1 Experimental method 2fTGH cells were treated with brecanatan at concentrations of 20, 40, 80 μM and HSV-1 respectively, DMSO+HSV-1 as control group, treated for 12 h; 2fTGH cells were treated with brecanatan at concentrations of 20, 40, 80 μM and EMCV respectively, DMSO+EMCV as control group, treated for 12 h; 2fTGH cells were treated with brecanatan at concentrations of 20, 40, 80 μM and MHV respectively, DMSO+MHV as control group, treated for 12 h; 2fTGH cells were treated with brecanatan at concentrations of 20, 40, 80 μM and IAV respectively, DMSO+IAV as control group, treated for 12 h, and the expression changes of viral genes were quantitatively detected by qPCR technology. In addition, the virus group and the DMSO group were used as controls to observe the effect of brecanatan on syncytium formation induced by HSV-1 and MHV in 2fTGH and J774.1 cells.

[0045] 2.2 Experimental results The results are shown in Figure 2 Specifically, the qPCR relative quantitative results show that brecanatan has an inhibitory effect on the transcription of HSV-1, EMCV, MHV and IAV viral genes, and the relative expression of viral genes decreases with the increase of drug concentration Figure 2 The cell syncytium experiment shows that the cell syncytium formation of the brecanatan treatment group is significantly reduced compared with the virus group and the DMSO group, indicating that the brecanatan drug can effectively inhibit the cell syncytium formation caused by MHV, HSV infection Figure 2 .

[0046] Therefore, brecanatan has broad-spectrum antiviral activity and can block syncytium induced by viruses, thereby reducing cell death caused by syncytium.

[0047] 3. Inhibition of VSV virus in other cells by brecanatan 3.1 Experimental method A549, HeLa, HT29, Pan02, Raw264.7 cells were treated with brecanatan (80 μM) and VSV, and the expression of viral genes was quantitatively analyzed by qPCR technology. In addition, Mock group and DMSO group were used as controls, and HeLa cells were treated with 30, 50 μM of brecanatan and VSV-GFP (green fluorescent protein) respectively, and the fluorescence intensity was observed by fluorescence microscope.

[0048] 3.2 Experimental results The results are shown inFigure 3 As shown in FIG. 13, specifically, the qPCR results show that the VSV viral mRNA expression level of the experimental group (betrixaban + VSV) is significantly lower than that of the control group (DMSO + VSV), thus it can be seen that betrixaban can significantly inhibit the transcription and expression of VSV in these cell lines. Figure 3 As shown in FIG. 13, specifically, the qPCR results show that the VSV viral mRNA expression level of the experimental group (betrixaban + VSV) is significantly lower than that of the control group (DMSO + VSV), thus it can be seen that betrixaban can significantly inhibit the transcription and expression of VSV in these cell lines. Figure 3 As shown in FIG. 13, specifically, the qPCR results show that the VSV viral mRNA expression level of the experimental group (betrixaban + VSV) is significantly lower than that of the control group (DMSO + VSV), thus it can be seen that betrixaban can significantly inhibit the transcription and expression of VSV in these cell lines.

[0049] 4. Mechanism of betrixaban antiviral activity 4.1 Experimental method The 2fTGH cells of cGAS knockout and wild type were treated with 20, 40, 80 μM of betrixaban and VSV respectively, and then the expression levels of antiviral genes MX1 and MX2 and the inhibition of VSV viral transcription were detected by qPCR.

[0050] 4.2 Experimental results As shown in FIG. 14, it can be seen from FIG. 14A that the expression of MX1 and MX2 genes in the cGAS knockout group is significantly lower than that in the betrixaban control group, indicating that the cGAS pathway is inhibited. Figure 4 As shown in FIG. 14, it can be seen from FIG. 14A that the expression of MX1 and MX2 genes in the cGAS knockout group is significantly lower than that in the betrixaban control group, indicating that the cGAS pathway is inhibited. Figure 4 As shown in FIG. 14, it can be seen from FIG. 14A that the expression of MX1 and MX2 genes in the cGAS knockout group is significantly lower than that in the betrixaban control group, indicating that the cGAS pathway is inhibited. Figure 4 As shown in FIG. 14, it can be seen from FIG. 14A that the expression of MX1 and MX2 genes in the cGAS knockout group is significantly lower than that in the betrixaban control group, indicating that the cGAS pathway is inhibited.

[0051] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application, and not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that they can modify the technical solutions described in the foregoing examples, or make equivalent substitutions for some technical features. The essence of the corresponding technical solution does not deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. Use of betrixaban or its pharmaceutically acceptable salts, solvates, hydrates, prodrugs, isomers, analogs, derivatives or metabolites in the preparation of antiviral drugs.

2. Use according to claim 1, characterized in that, The viruses include DNA viruses and / or RNA viruses.

3. Use according to claim 2, characterized in that, The DNA virus includes at least one of herpes simplex virus type 1, herpes simplex virus type 2, varicella-zoster virus, cytomegalovirus, smallpox virus, monkeypox virus, adenovirus, human papillomavirus, parvovirus, bocavirus, and hepatitis B virus.

4. Use according to claim 2, characterized in that, The RNA viruses include at least one of vesicular stomatitis virus, encephalomyocarditis virus, mouse hepatitis virus, influenza A virus, influenza B virus, influenza C virus, dengue virus, Zika virus, Ebola virus, Marburg virus, Nipah virus, coronavirus, hepatitis A virus, hepatitis C virus, rotavirus, measles virus, human immunodeficiency virus, respiratory syncytial virus, and rabies virus.

5. The use according to claim 1, characterized in that, The antiviral treatment includes at least one of the following: (1) Inhibit viral invasion of host cells; (2) Inhibit viral genome replication; (3) Inhibit viral protein processing or assembly; (4) Inhibit viral release; (5) Reduce the amount or activity of the virus.

6. Use according to claim 1, characterized in that, Antiviral effects are achieved by administering a therapeutically effective dose of the drug to the subject.

7. Use according to claim 6, characterized in that, The subjects included mammals.

8. The application according to claim 7, characterized in that, The mammals mentioned include humans.

9. The application according to claim 6, characterized in that, The effective therapeutic dose is 0.01-1000 mg / Kg.

10. Use of betrixaban or its pharmaceutically acceptable salts, solvates, hydrates, prodrugs, isomers, analogs, derivatives or metabolites in the preparation of combination antiviral drugs with other drugs.

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