CN113332279A

CN113332279A - Application of Ganetespib compound in preparation of anti-EV 71 virus drugs

Info

Publication number: CN113332279A
Application number: CN202110509496.5A
Authority: CN
Inventors: 魏艳红; 王海欣; 杨娜; 史玥玡; 徐佳慧; 罗芸; 胡康洪; 李涵洛
Original assignee: Hubei University of Technology
Current assignee: Hubei University of Technology
Priority date: 2021-05-11
Filing date: 2021-05-11
Publication date: 2021-09-03

Abstract

The invention discloses an application of a Ganetespib compound in preparation of an EV71 virus resistant drug. Through research experiments on the activity of Ganetespib against EV71, the compound Ganetespib (SAT-9090) inhibits cytopathic effect (CPE) generated by EV71 on host cells RD, enhances cell survival rate, reduces yield of progeny viruses, and has potential application in preparation of anti-EV 71 virus medicaments.

Description

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

Technical Field

The invention relates to the technical field of antiviral drugs, and in particular relates to an application of a Ganetespib compound in preparation of an anti-EV 71 virus drug.

Background

Enterovirus type 71 (EV71), a member of the Enterovirus genus (Enterovirus) of the Picornaviridae family (Picornaviridae), is one of the most prominent pathogens causing hand-foot-and-mouth disease in infants and young children, sometimes with severe central nervous system complications including aseptic meningitis, encephalitis, polio-like paralysis, neurological heart-lung failure, etc., and even death. The government of China has listed the hand-foot-and-mouth disease as a class C infectious disease in 2008 and brings into management, and a series of relevant laws and regulations are formulated to strictly control the epidemic spread of the hand-foot-and-mouth disease. There is currently no specific drug for the treatment of diseases infected by EV71, and related vaccines are marketed in 2015, and their preventive effects are yet to be further investigated. Therefore, the development of specific and effective anti-EV 71 medicaments is imperative.

Ganetespib (STA-9090) is an HSP90 inhibitor with IC in 8 cells of OSA₅₀4nM, induces apoptosis in OSA cells without affecting normal osteoblasts; is an active metabolite of STA-1474, currently in phase III of the clinic. STA-9090 was effective at nanomolar low concentrations in blocking cell proliferation and inducing apoptosis in a wide range of human tumor cell lines, and also had effects on many receptor tyrosine kinase inhibitors and tanesimycin-resistant cell lines. STA-9090 showed potent cytotoxicity in a range of solid and hematologic tumor cell lines, including those expressing mutant kinases resistant to small molecule tyrosine kinase inhibitors. Administration of STA-9090 resulted in significant tumor shrinkage in mice of several tumor xenograft models, and appeared to be less toxic. In addition, STA-9090 had better tumor permeability than tanespimicin. STA-9090 inhibited tumor growth in vivo in both malignant mast cell and OSA xenograft models. However, STA-9090 is rarely reported to be used for antiviral therapy.

Disclosure of Invention

In view of the defects of the prior art, the invention aims to evaluate the inhibition activity of STA-9090 on EV71 virus, and aims to provide the application of a Ganetespib compound in preparing an anti-EV 71 virus medicament, namely the STA-9090 is used as an effective EV71 virus inhibitor.

In order to achieve the purpose, the invention provides an application of a Ganetespib compound in preparing an anti-EV 71 virus medicine, which is characterized in that: the Ganetespib compound with anti-EV 71 activity, namely STA-9090, has the following chemical structural formula:

preferably, the Ganetespib compound inhibits EV71 by 85% at 6.25. mu.M.

Furthermore, the medicine refers to a preparation prepared by adding pharmaceutically acceptable auxiliary materials and carriers into the Ganetespib compound to resist EV71 virus.

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

The invention has the following advantages and beneficial effects:

the aim of the invention is realized by that STA-9090 tests the activity of the virus by a standard virus activity test method. According to the invention, a large number of biological experiments show that the STA-9090 has the activity of resisting the EV71 virus. The recombinant adenovirus 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 a great clinical application prospect.

Drawings

FIG. 1 is a graph of the effect of STA-9090 on RD cell survival by the action of EV 71.

FIG. 2 shows the inhibitory effect of STA-9090 on RD cell CPE caused by EV 71.

FIG. 3 shows the inhibitory effect of STA-9090 on the yield of EV71 progeny virus.

FIG. 4 shows the chemical structure of STA-9090.

Detailed Description

Ganetespib (STA-9090) used in the present invention was purchased from reagent Inc. The invention relates to application of Ganetespib (STA-9090) in preparation of an anti-EV 71 virus medicament, which is to add pharmaceutically acceptable auxiliary materials and carriers to Ganetespib (STA-9090) for preparing an anti-EV 71 virus preparation. The preparation is granule, tablet, pill, capsule, injection or dispersant.

The Ganetespib (STA-9090) has the following chemical structural formula.

Example 1

The application of the STA-9090 in resisting EV71 virus shows that the STA-9090 has an obvious effect in resisting EV71 virus. Therefore, the compound has potential to prepare specific therapeutic drugs for resisting EV71 infection and has a great clinical application prospect.

The invention carries out anti-EV 71 activity research experiments on the compounds, and the experimental conditions are as follows: hereinafter, materials and operation methods used in the present invention are well known in the art, if not specifically described.

1. The test contents are as follows:

analysis of compound anti-EV 71 activity: according to the invention, the STA-9090 anti-EV 71 activity is evaluated by combining a cytopathic effect analysis method and a MTT (methyl thiazolyl tetrazolium) determination cell survival rate detection method.

2. The test method comprises the following steps:

2.1.1 toxicity of Compounds on host RD cells

RD cells were plated in 96-well plates at 37 ℃ with 5% CO₂After the culture box is cultured to grow a monolayer, cell culture solution is discarded, cell maintenance solutions containing test compounds with different concentrations are respectively added for continuous culture, the cytotoxicity is visually observed and respectively recorded by a microscope after 48 hours, and the cell survival rate is measured by an MTT method. The SPSS 11.5 software calculates the Median cytotoxic concentration of the drug for the cells (CC 50). Cell survival rate ═ (mean OD of drug groups)₄₉₂Value/cell control mean OD₄₉₂Value) × 100%.

2.1.2 inhibitory Activity of Compounds on EV71

RD cells were plated in 96-well plates at 37 ℃ with 5% CO₂Culturing in incubator until monolayer grows, discarding culture solution, 100%The EV71 virus solution of TCID50 infected cells for 1.5h, and the cells were incubated with different concentrations of test compound (ribavirin as a positive control drug). After the culture is continued for about 48 hours, the cytopathic effect (CPE) is observed under a microscope when about 90% of CPE lesions appear in the virus control wells. Observation and recording method of CPE: no cytopathic effect is recorded as-below 25% cytopathic effect, 25% -50% cytopathic effect is recorded as +++, 50% -75% cytopathic effect is recorded as +++, and more than 75% cytopathic effect is recorded as ++++.

After CPE observation is finished, the inhibition rate of the drug on EV71 is detected by using an MTT method. The method comprises the following specific steps: MTT 50. mu.L (5 mg. multidot.mL) was added to each well^-1) After incubation for 3-4h, the supernatant was removed and an equal volume of DMSO was added to dissolve the pellet. The absorbance (OD) at 492nm was read with a microplate reader₄₉₂Value). The inhibition rate of the drug on EV71 was calculated using the following formula. The half effective Concentration of the drug (Concentration for 50% of maximum effect, EC50) was calculated using SPSS 11.5 software.

2.1.3 Therapeutic Index (TI) of drug

TI CC50/EC 50. A higher therapeutic index indicates greater antiviral potential.

3. Results of the experiment

TABLE 1 STA-9090 cytotoxicity and anti-EV 71 Activity

The results of the compound cytotoxicity and anti-EV 71 activity tests are shown in table 1. The effect of concentration-dependent compounds on RD cell viability with EV71 action is shown in figure 1. The maximal inhibition rate of STA-9090 at 6.25. mu.M was found to be about 85%.

Example 2

The invention carries out deep research on the activity of STA-9090 against EV71, and implements an inhibition effect test of the compound on the yield of EV71 progeny virus, wherein the test conditions are as follows:

1. content of the experiment

Compounds were tested for inhibition of EV71 progeny virus production following EV71 infection of RD cells.

2. Test method

RD cells in logarithmic growth phase are plated on 24-well plates and 100TCID after full monolayer growth₅₀EV71 infected cells were incubated at 37 ℃ for 1.5h, virus fluid was removed, washed three times with PBS, and cell maintenance fluid containing 6.25. mu.M was added. Collecting cells and supernatant culture solution after 48h, freeze thawing and cracking at-20 deg.C and 37 deg.C for three times, and performing TCID₅₀Methods EV71 virus titers were determined.

3. Test results

As shown in FIG. 3, the compound-treated RD cells all showed a significant reduction in viral titer relative to the viral control, which was a 6.5log reduction relative to the viral control.

In conclusion, the STA-9090 has strong EV71 inhibiting activity, can inhibit RD cytopathic effect caused by EV71 virus, and can be prepared into a medicament for clinically and effectively resisting EV71 infection.

Claims

1. The application of a Ganetespib compound in preparing an anti-EV 71 virus medicine is characterized in that: the Ganetespib compound with anti-EV 71 activity, namely STA-9090, has the following chemical structural formula:

2. use of a Ganetespib compound according to claim 1 in the preparation of a medicament against EV71 virus, wherein: the Ganetespib compound inhibited EV71 at 6.25 μ M by 85%.

3. Use of a Ganetespib compound according to claim 1 or 2 in the manufacture of a medicament against EV71 virus, wherein: the medicine is a preparation for resisting EV71 virus, which is prepared by adding pharmaceutically acceptable auxiliary materials and carriers into a Ganetespib compound.

4. Use of a Ganetespib compound according to claim 3 in the preparation of a medicament against EV71 virus, wherein: the preparation is any one of granules, tablets, pills, capsules, injections or dispersing agents.