CN113181152B

CN113181152B - Application of Tiratricol compound in preparation of anti-EV 71 virus drugs

Info

Publication number: CN113181152B
Application number: CN202110571357.5A
Authority: CN
Inventors: 魏艳红; 胡达; 刘会会; 胡康洪; 李涵洛
Original assignee: Hubei University of Technology
Current assignee: Hubei University of Technology
Priority date: 2021-05-25
Filing date: 2021-05-25
Publication date: 2023-04-28
Anticipated expiration: 2041-05-25
Also published as: CN113181152A

Abstract

The invention discloses an application of a Tiratricol compound in preparation of an EV71 virus resistant drug. Through a research experiment of the activity of Tiratricol against EV71, the compound Tiratricol inhibits cytopathic effect (CPE) generated by EV71 on host cells RD, enhances the survival rate of cells, reduces the yield of progeny viruses, and has potential to be applied to the preparation of anti-EV 71 virus medicaments.

Description

Application of Tiratricol compound in preparation of anti-EV 71 virus drugs

Technical Field

The invention belongs to the technical field of antiviral drugs, and particularly relates to application of a Tiratricol compound in preparation of an EV71 virus resistant drug.

Background

Enterovirus type 71 (EV 71) is a member of the Picornaviridae Enterovirus genus (Enterovirus), one of the most predominant pathogens causing infant hand-foot-and-mouth disease, sometimes accompanied by serious central nervous system complications including aseptic meningitis, encephalitis, poliomyelitis-like paralysis, neuropathic heart-lung failure, etc., and even leading to death. In view of the great harm to the life and health of people in China caused by the spreading and popularity of the hand-foot-and-mouth disease, the government in China has listed the hand-foot-and-mouth disease as a type-C infectious disease to be brought into management in 2008, and a series of relevant laws and regulations are formulated to strictly control the spreading and popularity of the hand-foot-and-mouth disease. At present, no specific medicine is available for treating the EV 71-infected diseases, and related vaccines are marketed in 2015, and the prevention effect of the vaccines is still to be further investigated. Therefore, development of specific and effective anti-EV 71 drugs is imperative.

Tiratricol (also known as TRIAC or triiodothyroacetic acid) is a thyroid hormone analog. Tiratricol is used for the treatment of thyroid hormone resistance syndrome, in combination with thyroxine for inhibiting the production of thyroid stimulating hormone in patients with thyroid cancer. It was studied for reducing goiter. Tiratricol also has a role in reducing atrophy caused by corticosteroid hormone use. However, tiratricol has been rarely reported for antiviral treatment.

Disclosure of Invention

In view of the above-mentioned current situation, the present invention aims to evaluate the inhibitory activity of Tiratricol on EV71 virus, and aims to provide an application of a thyroid hormone analogue Tiratricol compound in preparing anti-EV 71 virus drugs, namely, tiratricol as an anti-EV 71 virus screening compound.

In order to achieve the above purpose, the invention provides an application of a Tiratricol compound in preparing an anti-EV 71 virus drug, which is characterized in that: the Tiratricol compound has EV 71-resistant activity, and the chemical structural formula is shown as follows:

preferably, the inhibition of EV71 by the Tiratricol compound is approximately 100% at 25. Mu.M.

Further, the application refers to that the Tiratricol compound is added with pharmaceutically acceptable auxiliary materials and carriers for preparing an EV71 virus-resistant preparation.

Still further, the preparation is any one of granules, tablets, pills, capsules, injections or dispersing agents.

The object of the invention is achieved in that Tiratricol is tested for its activity by standard viral activity testing methods.

The invention has the advantages and beneficial effects as follows:

through a large number of biological experiments, tiratricol is found to have the activity of resisting EV71 virus. The specific expression is that the method can inhibit cytopathic effect caused by EV71 virus, enhance the survival rate of infected cells, inhibit the replication and proliferation of EV71 virus in cells and reduce the yield of progeny virus. Therefore, the compound has potential to prepare specific therapeutic drugs for resisting EV71 infection, and has great clinical application prospect.

Drawings

FIG. 1 is the effect of Tiratricol on RD cell viability of EV71 action.

FIG. 2 is the inhibitory effect of Tiratricol on EV71 induced RD cell CPE.

FIG. 3 is the inhibition of EV71 progeny virus production by Tiratricol.

Fig. 4 is a chemical structural diagram of Tiratricol.

Detailed Description

The technical scheme of the invention is further elaborated below with reference to specific embodiments and drawings.

Tiratricol used in the present invention was obtained from reagent company.

The invention relates to application of Tiratricol in preparation of anti-EV 71 virus medicaments.

The application refers to that the Tiratricol is added with pharmaceutically acceptable auxiliary materials and carriers to prepare an EV71 virus-resistant preparation.

The preparation is granule, tablet, pill, capsule, injection or dispersing agent.

The Tiratricol of the present invention has the following chemical structural formula.

The application of Tiratricol as the anti-EV 71 virus provided by the invention discovers that the effect of Tiratricol on resisting EV71 virus is obvious. Therefore, the compound has potential to prepare specific therapeutic drugs for resisting EV71 infection, and has great clinical application prospect.

Example 1

In this example, an anti-EV 71 activity study was performed on the above-mentioned compounds, and the experimental conditions were as follows: hereinafter, the materials and methods of operation used in the present invention are well known in the art, unless specifically indicated.

1. The test contents are as follows:

compound anti-EV 71 activity assay: the invention combines cytopathic effect analysis and MTT assay cell viability detection methods to evaluate the activity of Tiratricol against EV 71.

2. The test method comprises the following steps:

2.1.1 toxicity of Compounds to host RD cells

RD cells were plated in 96-well plates at 37℃with 5% CO ₂ Incubator culture lengthAfter the monolayer is full, the cell culture solution is discarded, cell maintenance solutions containing test compounds with different concentrations are respectively added for continuous culture, after 48 hours, the microscope is used for visual inspection, cytotoxicity is respectively recorded, and the cell survival rate is measured by an MTT method. SPSS 11.5 software calculated the median toxic concentration of drug to cells (Median cyctoxic concentration, CC 50). Cell viability= (mean OD of drug group ₄₉₂ Value/average OD of cell control group ₄₉₂ Value) x 100%.

2.1.2 inhibitory Activity of Compounds on EV71

RD cells were plated in 96-well plates at 37℃with 5% CO ₂ After the culture box is fully grown with a monolayer, the culture solution is discarded, the cells are infected by EV71 virus solution with 100TCID50 for 1.5 hours, and the cells are incubated by adding test compounds (ribavirin is used as a positive control drug) with different concentrations. When about 90% of CPE lesions appear in the virus control wells after further incubation for about 48 hours, cytopathic effects (CPE) are observed under a microscope. Observation and recording method of CPE: no cytopathy is marked as-, less than 25% of cytopathy is marked as +25% -50% of cytopathy is marked as++, 50% -75% of cytopathy is marked as++, and more than 75% of cytopathy is marked as++.

After the CPE is observed, the MTT method is used for detecting the inhibition rate of the drug to EV 71. The method comprises the following specific steps: MTT 50. Mu.L (5 mg. Multidot.mL) was added to each well ^-1 ) After 3-4h incubation, the supernatant was removed and the pellet was dissolved by adding an equal volume of DMSO. The absorbance (OD) corresponding to the absorbance was read at 492nm by using a microplate reader ₄₉₂ Values). The inhibition of EV71 by the drug was calculated using the following formula.

The half-effective concentration of the drug was calculated using SPSS 11.5 software (Concentration for 50%ofmaximal effect,EC50).

2.1.3 Therapeutic Index (TI) of the medicament

Ti=cc 50/EC50. The higher the therapeutic index, the greater the antiviral potential.

3. Experimental results

TABLE 1Tiratricol cytotoxicity and anti-EV 71 Activity

The cytotoxicity and anti-EV 71 activity test results of the compounds are shown in Table 1. The effect of concentration-dependent compound (Tiratricol) on EV 71-acting RD cell viability is shown in fig. 1, and the inhibitory effect of concentration-dependent compound (Tiratricol) on EV 71-induced RD cell CPE is shown in fig. 2. The maximum inhibition of Tiratricol was found to be close to 100% at 25 μm.

Example 2

In this example, the Tiratricol was studied in depth for anti-EV 71 activity, and an inhibition test of the EV71 progeny virus yield by the compound was performed as follows:

1. test content

Inhibition of EV71 progeny virus production by compounds after EV71 infection of RD cells was examined.

2. Test method

RD cells in log phase were plated in 24 well plates, 100TCID after confluence with monolayer ₅₀ EV71 infected cells were incubated at 37℃for 1.5h, the virus solution was removed, washed three times with PBS, and a cell-retaining solution containing a concentration of 25. Mu.M was added. Collecting cell and supernatant culture solution after 48h, and performing freeze thawing and lysis at-20deg.C and 37deg.C for three times to obtain TCID ₅₀ The method determines EV71 virus titer.

3. Test results

As shown in fig. 3, the compound treated RD cells had a significant decrease in viral titer relative to the viral control, which was more than 5.3log decrease relative to the viral control.

In conclusion, tiratricol has strong EV71 inhibition activity, can inhibit RD cytopathic effect caused by EV71 virus, and can be prepared into a medicament for effectively resisting EV71 infection clinically.

Claims

1. An application of a Tiratricol compound in preparing an anti-EV 71 virus medicament is characterized in that: the Tiratricol compound has EV 71-resistant activity, and the chemical structural formula is shown as follows:

2. use of a Tiratricol compound according to claim 1 for the preparation of an anti-EV 71 virus medicament, characterized in that: the inhibition of EV71 by the Tiratricol compound was nearly 100% at 25. Mu.M.

3. Use of a Tiratricol compound according to claim 1 or 2 for the preparation of an anti-EV 71 virus medicament, characterized in that: the application refers to that the Tiratricol compound is added with pharmaceutically acceptable auxiliary materials and carriers to prepare an EV71 virus-resistant preparation.

4. Use of Tiratricol as an EV71 virus inhibitor according to claim 3, characterized in that: the preparation is any one of granules, tablets, pills, capsules, injections or dispersing agents.