CN108210496B - Aromatic ester compound is used to prepare anti-ADV viral inhibitors - Google Patents

Abstract

The invention discloses aromatic ester compounds for preparing anti-ADV virus inhibitors. The aromatic ester compounds have the following chemical structural formula: compound of. Through the research on the anti-ADV activity of aromatic ester compounds WY113, WY130 and WY143, it was confirmed that aromatic ester compounds WY113, WY130 and WY143 can inhibit the cytopathic effect (CPE) of ADV on host cell RD, enhance cell survival rate, and reduce progeny virus Yield, inhibiting host cell apoptosis caused by ADV infection, and can be used to prepare anti-ADV virus inhibitors.

Description

Aromatic ester compounds are used to prepare anti-ADV virus inhibitors

technical field

The invention relates to the technical field of antiviral drugs, in particular to aromatic ester compounds WY113, WY130 and WY143 used to prepare anti-ADV virus inhibitors.

Background technique

Adenovirus (ADV) is a DNA virus isolated and cultured from surgically resected tonsil tissue. It mainly reproduces in the nucleus and often causes infection of human upper respiratory tract and eye epithelial cells. As a common opportunistic pathogen, adenovirus exists in the population for a long time, and patients with low immune function have a higher probability of adenovirus infection. As a non-enveloped DNA virus, adenovirus can exist stably at low pH and has strong resistance to physical and chemical agents. Adenovirus can produce tolerance to gastrointestinal secretions and bile, and can Replicates internally and produces a high viral load.

Adenoviruses can be transmitted from person to person through respiratory secretions, fecal-oral route and dirt. Many infants have been infected with at least one adenovirus within 5 years after birth, and this infection often occurs in densely populated places. , Adenovirus infection can occur at any time of the year, but outbreaks are usually concentrated in winter, spring and early summer. Adenoviruses often multiply in the pharynx, conjunctiva, intestinal tract, and lymphoid tissues, and cause a variety of clinical symptoms, such as respiratory infection, conjunctivitis, gastroenteritis, hepatitis, hemorrhagic cystitis, and nervous system disorders Wait. Adenovirus has attracted more and more attention because of its multiple types and wide range of pathogenicity. Therefore, it is imperative to develop a new anti-adenovirus drug.

Esters are an important class of fine chemical products, widely used in pharmaceuticals, materials, food, plasticizers, solvents and other chemical industries.

Contents of the invention

The object of the present invention is aimed at above-mentioned present situation, aims to provide aromatic ester compound to be used for preparing anti-ADV virus inhibitor.

The way to achieve the object of the present invention is that the aromatic ester compound is used to prepare anti-ADV virus inhibitors, and the aromatic ester compound is the following chemical structural formula:

When the concentration of aromatic ester compounds WY113, WY130 and WY143 is 40 μg/mL, the inhibitory rates of ADV-induced cytopathic effects are 94%, 95%, and 22%, respectively; they are used to prepare anti-ADV virus inhibitors.

Through a large number of biological experiments, the applicant found that aromatic ester compounds WY113, WY130 and WY143 have anti-ADV activity. The specific performance is that it can strongly inhibit the cytopathic effect caused by ADV virus and enhance the survival rate of infected cells. Strongly inhibit the replication and proliferation of ADV virus in cells, reduce the production of progeny virus, and protect cells from apoptosis caused by ADV infection. This shows that the aromatic ester compounds WY113, WY130 and WY143 have the potential to be used to prepare specific therapeutic drugs against ADV infection, and have great clinical application prospects.

The present invention has the following advantages:

1. The synthesis process of aromatic ester compounds WY113, WY130 and WY143 is simple, economical and fast, and easy for large-scale production and promotion.

2. Searching for anti-ADV drugs from compounds similar in structure to the aromatic ester compounds WY113, WY130 and WY143, it is easy to find their targets through structure-activity relationship research, and provide valuable guidance for further preparation and development of drugs.

Description of drawings

Figure 1 is a graph showing the effect of aromatic ester compounds WY113, WY130 and WY143 on the survival rate of RD cells under the action of ADV.

Fig. 2a, b, c are graphs showing the inhibitory effects of RD, ADV and ADV+WY143 on the apoptosis of RD cells induced by ADV, respectively.

Detailed ways

The applicant independently synthesized aromatic ester compounds WY113, WY130 and WY143 with novel structures, and disclosed the preparation methods of these aromatic ester compounds in the journal Tetrahedron Letters in 2015, but did not evaluate their biological activity.

The preparation method specifically uses the transition metal palladium as a catalyst, and under the induction of the ortho position of pyridine, hypervalent iodobenzene acts on the ortho position of the aromatic ring to perform aroyloxylation to obtain the final aromatic ester compound.

Aryl ester compounds WY113, WY130 and WY143 can strongly inhibit the cytopathic effect (CPE) of ADV in host cell RD and enhance cell survival rate. Strongly inhibit the replication and proliferation of ADV virus in cells, reduce the production of progeny virus, and protect cells from apoptosis caused by ADV infection. It shows that the aromatic ester compounds WY113, WY130 and WY143 can be used to prepare drugs for effectively treating anti-ADV infection and anti-ADV virus inhibitors.

The prepared anti-ADV virus inhibitor is granule, tablet, pill, capsule, injection, suspension or emulsion.

The applicant has conducted research experiments on the anti-ADV activity of the aromatic ester compounds WY113, WY130 and WY143, and the experimental conditions are as follows: In the following, unless otherwise specified, the materials and operating methods used in the present invention are well known in the art.

1. Test content:

Analysis of anti-ADV activity of compounds: In this invention, the anti-ADV activity of aromatic ester compounds WY113, WY130 and WY143 with novel structures will be evaluated by combining cytopathic effect analysis and MTT determination of cell survival rate detection methods.

2. Test method:

2.1.1 Toxicity of aromatic ester compounds WY113, WY130 and WY143 to host RD cells

Plate the RD cells on a 96-well plate, culture in a 37°C, 5% _CO2 incubator to cover the monolayer, discard the cell culture medium, and add the cell maintenance medium containing different concentrations of aromatic ester compounds WY113, WY130 and WY143 to continue the culture , After 48h, the cytotoxicity was visually observed under the microscope and recorded respectively, and the cell viability was determined by MTT method. SPSS 11.5 software calculates the median toxic concentration (Median cyctoxic concentration, CC50) of the drug on the cells. Cell survival rate=(average OD ₄₉₂ value of drug group/average OD ₄₉₂ value of cell control group)×100%.

2.1.2 Inhibitory activity of aromatic ester compounds WY113, WY130 and WY143 on ADV

The RD cells were plated on a 96-well plate, cultured in a 5% CO ₂ incubator at 37°C, and the culture medium was discarded, and the cells were infected with 100 TCID50 of the ADV virus liquid for 1 hour, and different concentrations of aromatic ester compounds WY113, WY130 and Cells were incubated with WY143 (ribavirin as positive control drug). After continuing to culture for about 48 hours, when about 90% of the CPE lesions appeared in the virus control wells, the cytopathic effect (CPE) was observed under a microscope. Observation and recording methods of CPE: No cytopathic changes are marked as -, cytopathic changes below 25% are marked as +, 25%-50% cytopathic changes are marked as ++, 50%-75% cytopathic changes are marked as +++, and more than 75% are marked as +++ Cytopathy was recorded as ++++.

After the observation of CPE, the inhibitory rate of drugs on ADV was detected by MTT method. The specific steps are: add 50 μL (5 mg·mL ^-1 ) of MTT to each well, remove the supernatant after incubation for 3-4 hours, and add an equal volume of DMSO to dissolve the precipitate. Read the corresponding absorbance (OD ₄₉₂ value) at 492 nm with a microplate reader. Use the following formula to calculate the inhibitory rate of drugs on ADV. The half effective concentration (Concentration for 50% of maximal effect, EC50) of the drug was calculated by SPSS 11.5 software.

2.1.3 The therapeutic index (SI) of the drug

SI=CC50/EC50. The higher the therapeutic index, the greater the antiviral potential.

3. The cytotoxicity and anti-ADV activity test results of aromatic ester compounds WY113, WY130 and WY143 are shown in Table 1. The effects of aromatic ester compounds WY113, WY130 and WY143 on the survival rate of ADV-treated RD cells are shown in Figure 1.

Table 1 Cytotoxicity and anti-ADV activity of aromatic ester compounds

The present invention detects that aromatic ester compounds WY113, WY130 and WY143 have strong inhibitory activity on ADV. Among them, the aromatic ester compound WY130 has a better inhibitory effect, and the aromatic ester compound WY143 has a decreased anti-ADV activity, but the toxicity is also significantly reduced, so the aromatic ester compounds WY130 and WY143 have a higher and similar therapeutic index. Aryl ester compounds WY113, WY130 and WY143 inhibit the CPE effect of RD cells induced by ADV as shown in Figure 2. ADV-infected RD cells became round and detached from the cell wall. Aryl ester compounds WY113, WY130 and WY143 (40 μ g/mL) had a certain inhibitory effect on the lesion effect. Aryl ester compound WY130 had a strong inhibitory effect and could almost Completely inhibit the RD cell pathological effect caused by ADV, and the inhibition rate reaches 96%.

The applicant further implemented the aromatic ester compounds WY113, WY130 and WY143 for the inhibition of RD cell apoptosis caused by ADV. The test conditions are as follows:

1. Test content

After ADV infects RD cells, it proliferates in the cells and destroys the normal life activities of cells, eventually leading to cell apoptosis. Therefore, the applicant further tested the inhibitory effect of aromatic ester compound WY143 on the apoptosis of RD cells induced by ADV after ADV infected RD cells.

2. Test method

The RD cells in the logarithmic growth phase were plated on 24-well plates, and the cells were infected with 100TCID50ADV after the monolayer was overgrown. After incubation at 37°C for 1.5h, the virus solution was removed, and the cell maintenance solution containing 40μg/mL aromatic ester compound WY143 was added. After about 48 hours, the cells were collected, and the apoptosis was detected on a flow cytometer using the Annexin V-FITC/PI apoptosis detection kit.

3. Test results

The experimental results showed that 40μg/mL aromatic ester compound WY143 could effectively inhibit the apoptosis caused by ADV. When the apoptosis rate of the virus control group was 97.6% (Figure 2-b), and the apoptosis rate of normal untreated cells was 0.59% (Figure 2-a), the apoptosis rate of the cells treated with 40 μg/mL aromatic ester compound WY143 and survival rates were 12.5% and 62.5%, respectively (Fig. 2-c). It can be seen that the aromatic ester compound WY143 can effectively protect cells from apoptosis caused by ADV.

Claims (3)

1. aromatic ester compound is preparing the application in anti-ADV viral inhibitors, it is characterised in that: aromatic ester compound is Following chemical structural formula:

Compound, fragrant ester compounds WY113, WY130 and WY143 concentration be 40 μ g/mL when, cytopathy is caused for ADV The inhibiting rate of change effect is respectively 94%, 95%, 22%；It is used to prepare anti-ADV viral inhibitors.

2. aromatic ester compound according to claim 1 exists preparing the application in anti-ADV viral inhibitors, feature In: fragrant ester compounds WY113, WY130 and WY143 add pharmaceutically acceptable auxiliary material and carrier, make by conventional method Standby anti-ADV viral inhibitors.

3. aromatic ester compound according to claim 1 exists preparing the application in anti-ADV viral inhibitors, feature In: the anti-ADV viral inhibitors of preparation are granule, tablet, pill, capsule, injection, suspending agent or emulsion.