Application of small molecule compound in preparation of drug for resisting dengue virus type II
By developing the small molecule compound ZP-96, the problem of lack of specific anti-DENV drugs in the prior art was solved, effective inhibition and low cytotoxicity of DENV-2 were achieved, and the potential of novel anti-dengue drugs was demonstrated.
Patent Information
- Application Number
- CN202510975768.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-15
- Publication Date
- 2025-09-02
AI Technical Summary
There is currently a lack of specific anti-DENV drugs approved by the FDA. The existing treatment methods mainly rely on symptomatic supportive therapy and cannot effectively control the dengue fever epidemic.
A small molecule compound ZP-96 was developed to prepare anti-dengue virus drug. Experiments have shown that it has strong inhibitory activity against DENV-2 in vitro and has low cytotoxicity. It is suitable for a variety of dosage forms such as capsules, pills, powders, tablets, oral liquids and injections.
ZP-96 showed significant anti-DENV-2 activity and low cytotoxicity, and had the potential to develop as a novel anti-dengue drug, which could effectively inhibit virus-induced cytopathy.
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Figure CN120570892A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of biomedicine and relates to the application of a small molecule compound in the preparation of an anti-dengue virus type II agent. Background Art
[0002] Dengue virus (DENV) belongs to the genus Flavivirus of the family Flaviviridae. It is a pathogen with an envelope structure and a single-stranded positive-strand RNA genome. It can be divided into four subtypes, DENV-1 to DENV-4, based on serotype differences, of which DENV-2 is the most commonly used subtype in research. DENV infection can cause dengue fever and the more severe dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Typical dengue fever patients will experience fever, severe headache, eye pain, muscle pain and bone and joint pain, fatigue, nausea, vomiting, diarrhea, etc., while patients with severe symptoms may experience symptoms such as severe bleeding and shock, or even death. The virus's unique antibody-dependent enhancement effect (ADE effect) greatly increases the risk of severe dengue fever (DSS / DHF). DENV is transmitted by the Aedes aegypti mosquito ( Aedes aegypti ) and Aedes albopictus ( Aedes albopictus ) is widely spread around the world and has now covered 128 countries. Epidemiological models predict that about 390 million people are infected each year, of which 96 million cases develop clinical symptoms, which has seriously endangered public health security.
[0003] Current clinical treatment is limited to symptomatic supportive care, such as fluid replacement to maintain electrolyte balance. Although international research has explored various strategies for anti-DENV drug development, existing prevention and treatment technologies are limited. Currently, there are no FDA-approved specific anti-DENV drugs, severely hampering global dengue fever prevention and control efforts. The development of anti-dengue virus drugs is urgently needed. Summary of the Invention
[0004] The present invention provides a new use of a small molecule compound ZP-96, namely, its use in the preparation of an anti-dengue virus type II drug. The structural formula of the small molecule compound ZP-96 is shown in Formula I: Formula I: ; The ingredients (or active ingredients) of the drug of the present invention are the above-mentioned small molecule compounds. One or more excipients acceptable in product production may also be added to improve the absorption effect of the active ingredients or facilitate use, such as capsules or pills, powders, tablets, granules, oral liquids and injections, that is, to prepare suitable dosage forms.
[0005] Compared with the prior art, the present invention has the following advantages and technical effects: The small molecule compound ZP-96 of the present invention is used in the treatment of dengue virus type 2. Experimental results show that it has strong anti-DENV-2 activity in vitro and low cytotoxicity. The present invention reveals that the compound has the potential to be developed into a new anti-dengue drug. BRIEF DESCRIPTION OF THE DRAWINGS
[0006] Figure 1 is the toxicity of different doses of ZP-96 on BHK-21 cells; Figure 2 The CC of ZP-96 on BHK-21 cells 50 Fitting curve graph; Figure 3 This is a graph showing that ZP-96 inhibits the cytopathic effect of DENV-2 infection in BHK-21 cells; Figure 4 ZP-96 is effective in the treatment of DENV-2 infected BHK-21 cells EC 50 Fitting curve graph; Figure 5 The CC of Ribavirin on BHK-21 cells 50 Fitting curve graph; Figure 6 Ribavirin inhibits DENV-2 infection in BHK-21 EC cells 50 Fitted curve graph. DETAILED DESCRIPTION
[0007] The present invention is further described in detail below with reference to the examples and accompanying drawings, but the embodiments of the present invention are not limited thereto. The scope of protection of the present invention is not limited to the contents described herein. The experimental methods used in these examples are conventional methods unless otherwise specified, and the materials and reagents used are commercially available unless otherwise specified. The DENV-2 described in the examples is DENV-2 (NGC).
[0008] The preparation of the small molecule compound ZP-96 in the following examples was prepared according to the method of Pan Zhou, Biao Hu, Lingling Lu, Rong Huang & Fuchao Yu. (2017). One-Pot Synthesis of Functionalised 4-Spiro-1,4-Dihydropyridines Via [1+2+1+2]-Cyclisation. Journal of Chemical Research (9), 513-516.
[0009] The small molecule compound ZP-96 powder in the following examples was dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution with a storage concentration of 50 mM. The completely dissolved drug was stored in a refrigerator at 4°C for subsequent experiments.
[0010] Example 1: Toxicity test of compound ZP-96 on BHK-21 cells 1. Take BHK-21 cells with good growth status, digest them with 0.25% trypsin, resuspend the cells and count the cells. 4 The cells were seeded at a density of 100 cells / well in a 96-well plate; 2. When the cells grow to more than 90%, dilute DMSO, compound ZP-96, and ribavirin in DMEM medium containing 2% FBS. Use 0.5% DMSO as a control group and treat the cells with different concentrations of ZP-96 or Ribavirin. 3. Culture in a 37°C, 5% CO2 incubator for 48 hours; 4. Microscope photography and recording; 5. Discard the culture medium, wash once with PBS, add 100 μL of MTT solution (1 mg / mL) to each well, and incubate in a 37°C, 5% CO2 incubator for 4 h; 6. After 4 h, discard the MTT, add 100 μL DMSO to each well to fully dissolve it, and measure the absorbance of each well at OD 490 nm on a microplate reader; See the results Figure 1 ,The results showed that 1000μM ZP-96 had very low toxicity to BHK-21 cells; Figure 2 The results showed that ZP-96 had an inhibitory effect on CC of BHK-21 cells. 50 >1000μM, Figure 5 The results showed that Ribavirin has an inhibitory effect on CC of BHK-21 cells. 50 It is 334μM.
[0011] Example 2: Compound ZP-96 inhibits DENV-2-induced cytopathic effects 1. Take BHK-21 cells with good growth status, digest them with 0.25% trypsin, resuspend the cells and count the cells. 4 The cells were seeded at a density of 100 cells / well in a 96-well plate; 2. When the cells grow to more than 90%, dilute the virus solution with DMEM medium containing 2% FBS and infect at an MOI of 1; 3. Incubate at 37°C, 5% CO2 incubator for 2 h. Dilute DMSO, compound ZP-96, and Ribavirin with DMEM medium containing 2% FBS. 0.2% DMSO was used as solvent control. The remaining groups were treated with different concentrations of ZP-96 or Ribavirin. 4. Culture in a 37°C, 5% CO2 incubator for 3 days; 5. The cell morphology of each group was recorded using a microscope in the compound ZP-96 treatment group; 6. Discard the maintenance solution, wash once with PBS, add 100 μL of MTT solution (1 mg / mL) to each well, and incubate in a 37°C, 5% CO2 incubator for 4 h; 7. After 4 hours, discard the MTT and add 100 μL of DMSO to each well to fully dissolve it. Measure the absorbance of each well at OD 490 nm on a microplate reader. See the results Figure 3 The results showed that compound ZP-96 at a concentration of 25 μM could significantly restore cell morphological changes caused by DENV-2; Figure 4 The results showed that ZP-96 could inhibit the cytopathic effect caused by DENV-2 in a dose-dependent manner. 50 18.71 μM; Figure 6 The results showed that Ribavirin effectively inhibited the cytopathic effect caused by DENV-2. 50 It is 8.726μM.
Claims
1. Use of a small molecule compound in the preparation of an anti-dengue virus type II drug, characterized in that: The chemical structure of the small molecule compound is as follows: 。