Use of betrixaban or a pharmaceutically acceptable salt thereof

Abstract

The application discloses application of betrixaban or a pharmaceutically acceptable salt thereof. Through drug screening and in-vitro drug experiments, it is found that betrixaban or the pharmaceutically acceptable salt thereof can inhibit virus replication and infection ability, and can block virus-induced syncytia, thereby reducing cell death caused by syncytia. In addition, the betrixaban or the pharmaceutically acceptable salt thereof of the application has broad-spectrum antiviral activity, and has the characteristics of safety and high efficiency.

Description

[0001] Cross-references 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, according to the application described in the present invention, 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.

[0010] In some embodiments, according to the application described in the invention, antiviral action is achieved by administering a therapeutically effective amount of the drug to the subject.

[0011] In some embodiments, according to the application described in the invention, the subject includes a mammal.

[0012] In some embodiments, according to the application described in the invention, the mammal includes a human.

[0013] In some embodiments, according to the application described in the invention, the therapeutically effective dose is 0.01-1000 mg / kg.

[0014] A second 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 a combination antiviral drug with other drugs.

[0015] This invention, through drug screening and in vitro drug experiments, has found that betraxaban, or its pharmaceutically acceptable salts, solvates, hydrates, prodrugs, isomers, analogs, derivatives, or metabolites, can inhibit viral replication and infectivity, and can block virus-induced syncytiotropium, thereby reducing syncytiotropium-induced cell death. Furthermore, betraxaban, or its pharmaceutically acceptable salts, solvates, hydrates, prodrugs, isomers, analogs, derivatives, or metabolites, exhibit broad-spectrum antiviral activity and are characterized by safety and high efficacy. Attached Figure Description

[0016] Figure 1 This study demonstrates the inhibitory effect of betrixaban on vesicular stomatitis virus (VSV) in human fibrosarcoma cells (2fTGH cells).

[0017] Figure 2 This study demonstrated the inhibitory effects of betrixaban on the transcription of other viruses and on syncytial formation.

[0018] Figure 3 The inhibitory effect of betrixaban on VSV virus in other cells was demonstrated.

[0019] Figure 4 The results of research on the antiviral mechanism of betrixaban are presented. Detailed Implementation

[0020] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0021] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that the upper and lower limits of the range and each intermediate value between them are specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0022] Unless otherwise stated, 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 invention pertains. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0023] 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 in the preparation of an antiviral medicament.

[0024] As used herein, the term "pharmaceutically acceptable salt" refers to both organic and inorganic salts of the compound betrixaban of this invention. Pharmaceutically acceptable salts are well known in the art, and examples include, but are not limited to: inorganic acid salts, such as hydrochloride, hydrobromide, phosphate, sulfate, and perchlorate; organic acid salts, such as acetate, oxalate, maleate, tartrate, citrate, succinate, and malonate; or salts obtained by other methods described in the literature, such as ion exchange. Other pharmaceutically acceptable salts include: adipate, malate, 2-hydroxypropionate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, transbutenedioic acid, gluconate, glycerophosphate, gluconate, hemisulfate, heptaate, hexanoate, hydroiodate, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pyrate, pectinate, persulfate, 3-phenylpropionate, picrate, pentanoate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0025] As used herein, the term "solvent" refers to an association or complex formed by one or more solvent molecules with the compounds of this invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol, etc.

[0026] As used herein, the term "hydrate" refers to an associative compound or complex formed with the compounds of this invention when the solvent molecule is water.

[0027] As used herein, the term "isomer" includes enantiomers, diastereomers, and geometric (or conformational) isomers of a given structure. For example, this invention includes R and S configurations for each asymmetry center, Z and E double bond isomers, Z and E conformational isomers, single stereochemical isomers, and mixtures of enantiomers, diastereomers, and geometric (or conformational) isomers. Unless otherwise stated, this invention includes all tautomers of the structures disclosed in Bétrixaban.

[0028] As used in this article, the term "prodrug" refers to a compound that is converted in vivo into a compound of formula (I) or (II): Formula (I); Equation (II).

[0029] The term "prodrug" refers to a parent drug molecule that is a pharmacologically inactive derivative that requires spontaneous or enzymatic biotransformation in vivo to release the active drug. Prodrugs are variants or derivatives of the compounds of this invention that have cleavable groups under metabolic conditions. When prodrugs undergo solvent degradation or enzymatic degradation under physiological conditions, they become pharmaceutically active compounds of this invention in vivo. Prodrug forms generally offer advantages such as 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, such as esters prepared by reacting a parent acid with a suitable alcohol, or amides prepared by reacting a parent acid compound with an amine, or with a basic group that reacts to form an acylated base derivative.

[0030] As used herein, the term "metabolite" refers to the product obtained in vivo by the metabolism of a specific compound or its salt. A metabolite of a compound can be identified using techniques known in the art, and its activity can be characterized by methods as known in the art. Such products can be obtained by subjecting the compound to oxidation, reduction, hydrolysis, acylation, deacylation, esterification, defatting, enzymatic cleavage, etc. Accordingly, the present invention includes metabolites of compounds of formula (I) or (II), including metabolites produced by sufficiently contacting compounds of formula (I) or (II) of the present invention with mammalian or cell lines derived therefrom for a period of time.

[0031] In this invention, the term "derivative" refers to a product derived from the substitution of atoms or functional groups in a compound of formula (I) or (II) by other atoms or functional groups. The derivatives are not specifically limited, including but not limited to esters, acylates, or metal complexes of compounds of formula (I) or (II).

[0032] In this invention, the virus includes DNA viruses and / or RNA viruses. Examples of the DNA viruses 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, and hepatitis B virus. Examples of the RNA viruses include, but are not limited to, vesicular stomatitis virus, encephalomyelitis 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. In one 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 encephalomyelitis virus. In yet another preferred embodiment, the RNA virus is mouse hepatitis virus. In yet another preferred embodiment, the RNA virus is an influenza A virus.

[0033] In this invention, "antiviral" refers to improving a condition before or after the onset of a disease or dysfunction. This degree of relief or prevention, measured by any standard technique, is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100% compared to an untreated control group under equivalent conditions. Beneficial or desired clinical outcomes include, but are not limited to, the following, whether detectable or undetectable: symptom relief, reduction in disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, and reduction (whether partial or complete). Antiviral actions include, but are not limited to, (1) inhibiting viral invasion of host cells; (2) inhibiting viral genome replication; (3) inhibiting viral protein processing or assembly; (4) inhibiting viral release; and (5) reducing viral quantity or activity.

[0034] In this invention, antiviral effects are achieved by administering a therapeutically effective amount of the drug to a subject. Subjects include, but are not limited to, mammals, including but not limited to, humans, mice, rabbits, cats, dogs, cattle, sheep, and pigs.

[0035] In this invention, the medicament contains a therapeutically effective amount of betraxaban or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, isomer, analog, derivative, or metabolite thereof, particularly betraxaban 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 medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, which participates in the transport or delivery of the medicament from one organ or part of the body to another organ or part of the body. Each excipient or carrier is "acceptable," meaning it is compatible with other components of the formulation and does not harm the patient.

[0036] In this invention, there are no particular limitations on the method of administration of the drug. Representative methods of administration include, but are not limited to, oral, rectal, parenteral (intravenous, intramuscular, or subcutaneous) and local administration. Accordingly, the drug of this invention can be formulated into various clinically acceptable dosage forms, including oral dosage forms, injectable dosage forms, local administration dosage forms, or topical dosage forms.

[0037] The therapeutically effective dose described in this invention refers to a pharmaceutically recognized effective dosage, that is, an amount of the active compound (i.e., betraxaban or its pharmaceutically acceptable salts, solvates, hydrates, prodrugs, isomers, analogs, derivatives or metabolites, especially betraxaban maleate) sufficient to significantly improve the condition without causing serious side effects. The daily dose of betraxaban or its pharmaceutically acceptable salts, solvates, hydrates, prodrugs, isomers, analogs, derivatives or metabolites is generally 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 dosages include, 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 dosage is calculated as betraxaban or its pharmaceutically acceptable salts, solvates, hydrates, prodrugs, isomers, analogs, derivatives, or metabolites. It can be administered as a single dose once daily, or divided into multiple doses daily, or used at intervals.

[0038] Combined applications One aspect of the present 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 solvent) 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 Methods Two-fTGH cells were treated with betrixaban and herpes simplex virus type 1 (HSV-1) at concentrations of 20, 40, and 80 μM, respectively, with DMSO+HSV-1 as the control group, for 12 h. Two-fTGH cells were treated with encephalomyocarditis virus (EMCV) at concentrations of 20, 40, and 80 μM, respectively, with DMSO+EMCV as the control group, for 12 h. Two-fTGH cells were treated with mouse hepatitis virus (MHV) at concentrations of 20, 40, and 80 μM, respectively, with DMSO+MHV as the control group, for 12 h. Two-fTGH cells were treated with influenza A virus (IAV) at concentrations of 20, 40, and 80 μM, respectively, with DMSO+IAV as the control group, for 12 h. Changes in viral gene expression were quantitatively detected using qPCR. In addition, using the virus group and DMSO group as controls, the effects of betrixaban on syncytial formation induced by HSV-1 and MHV in 2fTGH and J774.1 cells were observed.

[0045] 2.2 Experimental Results The results are as follows Figure 2 As shown, specifically, qPCR relative quantification results indicated that betraxaban inhibited the transcription of HSV-1, EMCV, MHV, and IAV viral genes, and the relative expression levels of viral genes decreased with increasing drug concentration. Figure 2 (A). Cell syncytial assays showed that, compared with the virus group and the DMSO group, the betraxaban treatment group exhibited a significant reduction in cell syncytial formation, indicating that betraxaban can effectively inhibit cell syncytial formation induced by MHV and HSV infections. Figure 2 (B).

[0046] Therefore, betrixaban has broad-spectrum antiviral activity and can block virus-induced syncytiocytosis, thereby reducing syncytiocytosis-induced cell death.

[0047] 3. Inhibitory effect of betrixaban on VSV virus in other cells 3.1 Experimental Methods A549, HeLa, HT29, Pan02, and Raw264.7 cells were treated with betrixaban (80 μM) and VSV, and viral gene expression was quantitatively analyzed using qPCR. In addition, the Mock and DMSO groups were used as controls. HeLa cells were treated with 30 and 50 μM betrixaban and VSV-GFP (green fluorescent protein), respectively, and the fluorescence intensity was observed using a fluorescence microscope.

[0048] 3.2 Experimental Results The results are as follows Figure 3 As shown, specifically, qPCR results indicated that the VSV virus mRNA expression level in the experimental group (betraxaban + VSV) was significantly lower than that in the control group (DMSO + VSV). This demonstrates that betraxaban can significantly inhibit the transcriptional expression of VSV in these cell lines. Figure 3 (A). Microscopic results showed that, compared with the control group, the number of green fluorescent cells decreased significantly with increasing betrixaban concentration, indicating that betrixaban can inhibit VSV-GFP expression (A). Figure 3 (B). In summary, betrixaban has significant antiviral activity and can inhibit viral replication in a variety of cell lines.

[0049] 4. Research on the antiviral mechanism of betrixaban 4.1 Experimental Methods cGAS knockout and wild-type 2fTGH cells were treated simultaneously with 20, 40, and 80 μM betrixaban and VSV, respectively. 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 The results are as follows Figure 4 As shown, by Figure 4 As shown in A, compared to the betrixaban control group, the expression of MX1 and MX2 genes in the cGAS knockout group was significantly reduced, indicating that the cGAS pathway was inhibited. Figure 4 As shown in B, the VSV mRNA gene detection results showed that in the wild-type 2fTGH model, compared with the DMSO control group, the expression of VSV mRNA in the betraxaban group was significantly decreased, and the expression level decreased with increasing drug concentration. Similarly, in the cGAS knockout model, the expression level of VSV mRNA in the betraxaban group decreased with increasing drug concentration. This indicates that betraxaban exerts its antiviral effect not only through the cGAS pathway, but also that cGAS is an important pathway among multiple antiviral pathways.

[0051] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The use of betrixaban in the preparation of antiviral drugs, characterized in that, The virus is vesicular stomatitis virus, herpes simplex virus type 1, encephalomyocarditis virus, or influenza A virus, and the drug further includes pharmaceutically acceptable excipients or carriers.

2. The application 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.

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

4. The application according to claim 3, characterized in that, The subjects included mammals.

5. The application according to claim 4, characterized in that, The mammals mentioned include humans.

6. The application according to claim 3, characterized in that, The effective therapeutic dose is 0.01-1000 mg / Kg.

Images