Use of anthraquinone compound aloe saponarin II in preparation of CVB3 virus inhibitor

Abstract

The application discloses application of an anthraquinone compound aloe saponarin II in preparation of a CVB3 virus inhibitor and belongs to the technical field of antiviral drugs. The application finds that the anthraquinone compound aloe saponarin II has the activity of resisting CVB3 virus, can inhibit the cytopathic effect caused by the CVB3 virus, enhance the survival rate of infected cells, inhibit the replication and proliferation of the CVB3 virus in cells and reduce the progeny virus yield. Thus, it is shown that the anthraquinone compound aloe saponarin II has the potential for being used for preparing a specific treatment drug against CVB3 infection and has a broad clinical application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of antiviral drugs, and in particular to application of an anthraquinone compound aloesaponarin II in the preparation of CVB3 virus inhibitors. Background Art

[0002] Coxsackieviruses are members of the Enterovirus genus within the Picornaviridae family. Infection with coxsackieviruses can cause a variety of diseases, including hand, foot, and mouth disease, aseptic meningitis, encephalitis, myocarditis, myositis, and herpangina. There are 29 reported serotypes of coxsackieviruses, which can be divided into two groups, A and B, based on their pathogenicity in suckling mice and their cellular sensitivities: CVA (CVA1–22, 24) and CVB (CVB1–6). Infection with CVBs is the most common, with CVB3 being the most pathogenic of the six CVB serotypes and the primary cause of viral myocarditis. Currently, there are no specific drugs for coxsackievirus infection, and no targeted clinical treatment is available. Therefore, the development of specific and effective anti-CVB3 drugs is imperative.

[0003] Anthraquinone compounds have a wide range of pharmacological activities and are often used as laxatives, anti-inflammatory drugs, and antibacterial drugs. They also have good efficacy in antiviral and immune regulation. Aloesaponarin II is a natural anthraquinone product that was first isolated from the fresh rhizomes of Aloe saponaria. Studies have shown that aloesaponarin II has significant cytotoxic activity against the human cervical cancer cell line KB-3-3, with IC 50 The value was 0.98 μM. However, the inhibitory activity of aloesaponarin II against CVB3 has not been reported yet. Summary of the Invention

[0004] The purpose of the present invention is to provide the use of an anthraquinone compound aloesaponarin II in the preparation of CVB3 virus inhibitors, so as to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides use of an anthraquinone compound aloesaponarin II in preparing a CVB3 virus inhibitor.

[0007] The anthraquinone compound aloesaponarin II has a structure shown in formula (I):

[0008] Formula (I).

[0009] The present invention also provides use of an anthraquinone compound aloesaponarin II in preparing medicines for treating diseases caused by CVB3 virus.

[0010] The present invention also provides a pharmaceutical composition for treating diseases caused by CVB3 virus, comprising the anthraquinone compound aloesaponarin II.

[0011] The present invention also provides a pharmaceutical preparation for treating diseases caused by CVB3 virus, comprising the pharmaceutical composition and a pharmaceutically acceptable carrier.

[0012] Based on the above technical solution, the present invention has the following technical effects:

[0013] The present invention discovered that the anthraquinone compound aloesaponarin II has anti-CVB3 virus activity, inhibiting the cytopathic effect caused by the CVB3 virus, enhancing the survival rate of infected cells, inhibiting the replication and proliferation of the CVB3 virus within cells, and reducing the production of progeny viruses. This indicates that the anthraquinone compound aloesaponarin II has the potential to be used in the preparation of a specific therapeutic drug against CVB3 infection and has broad clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0015] Figure 1 The inhibitory effect of the anthraquinone compound aloesaponarin II on CPE of Hep-2 cells induced by CVB3.

[0016] Figure 2 This is the effect of the anthraquinone compound aloesaponarin II on the survival rate of Hep-2 cells treated with CVB3.

[0017] Figure 3 The inhibitory effect of the anthraquinone compound aloesaponarin II on the production of CVB3 progeny virus. DETAILED DESCRIPTION

[0018] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0019] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0020] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0021] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the present invention. The present description and examples are intended to be illustrative only.

[0022] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0023] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or are publicly available.

[0024] The embodiments of the present invention provide the use of anthraquinone compounds in the preparation of CVB3 virus inhibitors.

[0025] In some specific embodiments, the anthraquinone compound is aloesaponarin II, and its structural formula is shown in formula (I):

[0026] Formula (I).

[0027] The embodiments of the present invention provide the use of anthraquinone compounds in the preparation of drugs for treating diseases caused by CVB3 virus.

[0028] In some specific embodiments, the anthraquinone compound is aloesaponarin II.

[0029] The application provides a pharmaceutical composition for treating diseases caused by CVB3 virus, comprising the anthraquinone compound aloesaponarin II.

[0030] The anthraquinone compound aloesaponarin II used in the application is obtained by the inventors from liquid fermentation extracts of Streptomyces tanashiensis DSM 731.

[0031] The application provides a pharmaceutical preparation for treating diseases caused by CVB3 virus, comprising the pharmaceutical composition and a pharmaceutically acceptable carrier.

[0032] In some specific embodiments, the pharmaceutically acceptable carrier is suitable for granules, tablets, pills, capsules, injections or dispersions.

[0033] Example 1

[0034] 1. Test content:

[0035] Compound anti-CVB3 activity analysis: the anti-CVB3 activity of the anthraquinone compound aloesaponarin II is evaluated by combining cytopathic effect analysis and MTT determination of cell survival rate detection method.

[0036] 2. Test method:

[0037] 2.1 Compound toxicity to host Hep-2 cells

[0038] Hep-2 cells are plated in a 96-well plate and incubated at 37°C in a 5% CO2 incubator until a monolayer is formed. The cell culture medium is then discarded and replaced with cell maintenance medium containing the anthraquinone compound aloesaponarin II at concentrations of 200 µM, 150 µM, 100 µM and 75 µM. After 48 hours, the cell toxicity is observed under a microscope and the cell survival rate is determined by MTT method. The Median cyctoxic concentration (CC 50 ) of the drug to the cells is calculated using SPSS 11.5 software.

[0039] Cell survival rate = (average OD 492 value of the drug group / average OD 492 value of the cell control group) × 100%.

[0040] 2.2 Inhibitory activity of the compound to CVB3

[0041] Hep-2 cells were plated in 96-well plates and cultured in a 37°C, 5% CO2 incubator until confluent monolayers were formed. The culture medium was discarded and the cells were infected with 100 TCID50 of CVB3 virus solution for 1.5 h. The cells were then incubated with aloesaponarin II at concentrations of 200 μM, 100 μM, 50 μM, 25 μM, and 12.5 μM. Ribavirin (concentrations of 500 μM, 250 μM, 125 μM, 62.5 μM, and 31.25 μM) was used as a positive control drug. After incubation for approximately 48 h, when approximately 90% of the virus control wells showed CPE, the cells were observed under a microscope. Figure 1 As shown. Hep-2 cells typically appear oval or polygonal and adhere to the culture dish. Viral infection can cause significant cytopathological effects in Hep-2 cells, with the cells becoming rounded and detaching from the culture dish. CPE is recorded as follows: no cytopathic effect is recorded as -, less than 25% cytopathic effect is recorded as +, 25%-50% cytopathic effect is recorded as ++, 50%-75% cytopathic effect is recorded as +++, and greater than 75% cytopathic effect is recorded as ++++.

[0042] After the CPE observation was completed, the MTT method was used to detect the inhibitory rate of drugs on CVB3. The specific steps were as follows: 50 μL of MTT (5 mg mL –1 After incubation for 3-4 h, the supernatant was removed and an equal volume of DMSO was added to dissolve the precipitate. The corresponding absorbance (OD) was read at 492 nm using a microplate reader. 492 The inhibition rate of drugs on CVB3 was calculated using the following formula. The concentration for 50% of maximal effect (EC) was calculated using SPSS 11.5 software. 50 ).

[0043] Virus inhibition rate = [OD of drug-treated group (drug + virus) 492 - Virus control group OD 492 ] / (OD of cell control group 492 - Virus control group OD 492 ) × 100%

[0044] 2.3 Therapeutic Index (TI) of a Drug

[0045] TI = CC 50 / EC 50 .

[0046] The higher the therapeutic index, the greater the antiviral potential.

[0047] 3. Experimental results

[0048] Table 1 Cytotoxicity and anti-CVB3 activity of anthraquinone compound aloesaponarin II

[0049]

[0050] The results of the cytotoxicity and anti-CVB3 activity tests of the anthraquinone compound aloesaponarin II are shown in Table 1. It has significant inhibitory activity against CVB3, and its therapeutic index exceeds that of the positive control drug ribavirin, indicating that it has great antiviral potential. Figure 2 As shown, the inhibition rate reached 95% at 25µM.

[0051] Example 2

[0052] The anthraquinone compound aloesaponarin II was tested for its inhibitory effect on the production of CVB3 progeny viruses. The test results are as follows:

[0053] 1. Test content

[0054] The inhibitory effect of the anthraquinone compound aloesaponarin II on the production of CVB3 progeny viruses after CVB3 infected Hep-2 cells was detected.

[0055] 2. Test methods

[0056] Hep-2 cells in the logarithmic growth phase were plated in 24-well plates. After the cells grew into a confluent monolayer, 100 TCID 50 The cells were infected with CVB3 and incubated at 37°C for 1.5 h, then the virus solution was removed, washed three times with PBS, and aloesaponarin II was added to maintain the cells. After 48 h, the cells and supernatant were collected and lysed by freezing and thawing three times at –20°C and 37°C. The TCID 50 Methods The CVB3 virus titer was determined.

[0057] 3. Test results

[0058] like Figure 3 As shown, the viral titer of Hep-2 cells treated with the anthraquinone compound aloesaponarin II was significantly reduced compared with the virus control group, with a decrease of 4.8 log relative to the virus control.

[0059] In summary, the anthraquinone compound aloesaponarin II has strong inhibitory activity against CVB3, can inhibit Hep-2 cell death caused by CVB3 virus, and can be prepared into a clinically effective drug against CVB3 infection.

[0060] Obviously, the above embodiments of the present invention are merely examples for the purpose of clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. A person skilled in the art will be able to make other variations or modifications based on the above description. It is not necessary and impossible to enumerate all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the claims of the present invention.

Claims

1. Use of an anthraquinone compound aloesaponarin II in the preparation of a drug for inhibiting CVB3 virus, characterized in that: The anthraquinone compound aloesaponarin II has a structure as shown in formula (I): Formula (I).

2. The use according to claim 1, characterized in that When the concentration of the anthraquinone compound aloesaponarin II is 25 μM, the inhibition rate of CVB3 reaches 95%.

3. The use according to claim 2, characterized in that: The anthraquinone compound aloesaponarin II is used to prepare a CVB3 virus inhibitor after adding pharmaceutically acceptable excipients.

4. The use according to claim 3, characterized in that: The preparation is any one of tablets, capsules, granules, pills or injections of common medicines.

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