Application of carbinoxamine maleate in preparation of anti-influenza virus medicine
阅读说明：本技术 马来酸卡比沙明在制备抗流感病毒药物中的应用 (Application of carbinoxamine maleate in preparation of anti-influenza virus medicine ) 是由 陆路 姜世勃 徐巍 刘腾 王茜 花晨 王聪 付玉红 李佩玉 于 2018-08-29 设计创作，主要内容包括：本发明属于医药技术领域,涉及马来酸卡比沙明在制备抗甲型流感病毒药物中的应用。经实验表明,所述马来酸卡比沙明在完全无毒性浓度时对不同型和亚型的流感病毒均有显著的抗流感活性,并呈剂量依赖性地抑制所有检测病毒株的复制,表明马来酸卡比沙明抗流感病毒活性具有一定的广谱性,本发明所述流感病毒包括甲型和乙型流感病毒,包括H1N1、H3N2、H7N9、H5N1、H7N1、H7N2、H7N3、H7N7、H9N2或B型；本发明作用机制研究表明马来酸卡比沙明主要通过干扰流感病毒的内吞作用抑制病毒进入宿主细胞；所述的马来酸卡比沙明可制备用于预防和治疗流感的药物,为预防和治疗流感提供新的途径和手段。(The invention belongs to the technical field of medicines, and relates to application of carbinoxamine maleate in preparation of an anti-influenza A virus medicine. Experiments show that the carbinoxamine maleate has obvious anti-influenza activity on different types and subtypes of influenza viruses when in completely nontoxic concentration, inhibits the replication of all detected virus strains in a dose-dependent manner, and shows that the anti-influenza virus activity of the carbinoxamine maleate has certain broad spectrum, wherein the influenza viruses comprise influenza A viruses and influenza B viruses, including H1N1, H3N2, H7N9, H5N1, H7N1, H7N2, H7N3, H7N7, H9N2 or B viruses; the research of the action mechanism of the invention shows that the carbinoxamine maleate mainly inhibits the virus from entering the host cell by interfering the endocytosis of the influenza virus; the carbinoxamine maleate can be used for preparing a medicine for preventing and treating influenza, and provides a new way and means for preventing and treating the influenza.)
1. The application of the compound of the maleic acid carbinoxamine in preparing the anti-influenza virus medicament,
2. the use according to claim 1, wherein the influenza virus is an influenza a virus or an influenza b virus.
3. Use according to claim 2, characterized in that: the influenza virus is type A H1N1, H3N2, H7N9, H5N1, H7N1, H7N2, H7N3, H7N7, H9N2 or type B.
4. Use of carbinoxamine maleate in the preparation of a medicament for inhibiting the entry of influenza virus into a target cell.
5. Use according to claim 1 or 2, characterized in that: the compound of the carbinoxamine maleate inhibits the virus from entering and has the function of resisting influenza virus with host cells through the targeted entry process of the influenza virus.
6. Use according to claim 1 or 2, characterized in that: the anti-influenza virus medicament is a medicament for preventing and treating influenza.
Technical Field
The invention belongs to the technical field of medicines, and relates to application of carbinoxamine maleate in preparation of an anti-influenza A virus medicine.
Background
Disclosure of Invention
The invention aims to provide a novel anti-influenza virus inhibitor aiming at the deficiency of the drugs for clinically treating influenza virus based on the current situation of the prior art, and provides a safe and efficient small molecular compound with small toxic and side effects for clinically treating influenza; in particular to an application of carbinoxamine maleate in preparing anti-influenza virus drugs.
The carbinoxamine maleate (CAM) can effectively inhibit the activity of influenza A and B viruses in a non-toxic range, and can be further developed into a medicament for treating influenza virus infection diseases, so that the carbinoxamine maleate has wider application prospect.
In the invention, an FDA approved drug library containing 1,280 compounds is screened, and an antihistamine drug carbinoxamine maleate (CAM) with a structure shown in a formula I is obtained, and experiments prove that the antihistaminic drug carbinoxamine maleate has strong inhibitory activity on influenza A virus strain A/Shanghai/4664T/2013(H7N 9);
the carbinoxamine maleate (CAM) (trade name: Palgic, Histex PD, Karbinal ER, Carboxin) is an antihistamine drug acting on H1 receptor, belongs to a small molecule compound which is artificially synthesized, is approved by the US FDA in 1950 to treat pollinosis, rhinitis, urticaria, asthma and the like, but no guidance for the application of CAM directly used for resisting influenza virus exists so far. According to the invention, the in vitro and in vivo antiviral activity of the CAM on different types and subtypes of influenza viruses is evaluated through experiments, particularly, the CAM can remarkably inhibit the infection of mice on influenza A virus strain H7N9, and the potential action mechanism of the CAM is researched, and the result shows that the CAM has the application of resisting the influenza viruses and has the prospect of becoming an anti-influenza virus medicament.
In the invention, the compound CAM can inhibit the replication of influenza A viruses H1N1, H3N2, H7N9 and influenza B viruses in host cells;
the compound CAM can target at the endocytosis stage of influenza virus, thereby inhibiting the virus from entering host cells, having obvious antiviral activity, and being used for developing anti-influenza drugs.
The compound CAM disclosed by the invention is a clinical medicine approved by FDA, and has the characteristics of safety, high efficiency and small toxic and side effects.
The invention provides a new application of carbinoxamine maleate in preparing anti-influenza virus drugs; the influenza virus comprises influenza A and B viruses, preferably H1N1, H3N2, H7N9, H5N1, H7N1, H7N2, H7N3, H7N7, H9N2 or B; the carbinoxamine maleate has obvious anti-influenza activity on different types and subtypes of influenza viruses when the concentration of the carbinoxamine maleate is completely free from toxicity, and inhibits the replication of a detected virus strain in a dose-dependent manner, and an experimental result shows that the anti-influenza virus activity of the carbinoxamine maleate has broad spectrum;
the invention carries out action mechanism research, and the result shows that the carbinoxamine maleate mainly inhibits the virus from entering host cells by interfering the endocytosis of the influenza virus, and the medicament for preventing and treating the influenza is prepared.
Furthermore, the carbinoxamine maleate can be used for preparing novel anti-influenza A and anti-influenza B virus drugs, and provides a new way and means for preventing and treating influenza.
The compound CAM can be prepared into various pharmaceutically acceptable dosage forms such as tablets, granules, powder, capsules, oral liquid, injection and the like by using a conventional method in the field of modern pharmacy, and is used for treating influenza.
Compared with the prior art, the invention has the following advantages and effects:
1. the compound CAM can inhibit the infection of influenza A virus strains H1N1, H3N2 and H7N9 and influenza B virus to cells, and has broad-spectrum antiviral activity.
2. Compound CAM is less toxic to cells, and has CC50 values of 240.52. + -. 12.53. mu.M for MDCK cells. IC of CAM against influenza A H1N1, H3N2, H7N9 strains and influenza B viruses50The anti-influenza virus anti-virus composition is different from 4.24 to 15.54 mu M, has a safety index of 15-68, and can be used as a safe and effective anti-influenza virus medicament.
3. Compound CAM acts on the initial entry phase of influenza virus, thereby inhibiting viral entry into the host cell. Meanwhile, the compound can solve the drug resistance aiming at the target M2 ion channel and neuraminidase, and is a potential effective antiviral drug.
Drawings
The chemical structure of the compound CAM of FIG. 1.
Figure 2 toxicity assay of compound CAM on MDCK cells.
Figure 3 activity of compound CAM against influenza virus, wherein, a. inhibitory activity of compound CAM against influenza a virus strain a/Shanghai/37T/2009(H1N 1); B. inhibitory activity of compound CAM against influenza A strain A/Puerto Rico/8/1934(H1N 1); C. inhibitory activity of compound CAM against influenza A strain A/Guizou/54/1989 (H3N 2); D. inhibitory Activity of Compound CAM against influenza B virus strain B/Shanghai/2017 (BY).
Figure 4, in vivo antiviral activity assay of compound CAM, wherein the effect of compound CAM on survival of mice infected with highly pathogenic influenza a
Figure 5, test of antiviral activity interfering with CAM at different time points after influenza infection, compound CAM acting early on influenza infection.
Figure 6, effect of compound CAM on viral adsorptive invasion, wherein, a. inhibitory activity of compound CAM on H7N9 pseudovirus infection; B. inhibitory Activity of Compound CAM against VSV pseudovirus infection.
Figure 7, effect of compound CAM on erythrocyte agglutination and progeny virus release, wherein, a. effect of compound CAM on erythrocyte agglutination; B. effect of compound CAM on neuraminidase activity.
Detailed Description
For a better understanding of the context of the present invention, the following experiments and results are combined to further illustrate the use of compound CAM against influenza A and B viruses.
The materials used in the present invention:
cell and virus: canine kidney epithelial cells (MDCK cells) and 293T cells were grown in DMEM medium containing 10% fetal bovine serum, 100U/L penicillin, streptomycin; influenza viruses A/Shanghai/37T/2009(H1N1), A/Puerto Rico/8/1934(H1N1), A/Guizhou/54/1989(H3N2), A/Shanghai/4664T/2013(H7N9) and B/Shanghai/2017(BY) were given away BY Shanghai City public health clinical center; plasmids H7N9HA, VSV-G and pNL4-3R-E-Luc were used for pseudovirus production; pNL4-3R-E-Luc is a recombinant plasmid that has been frameshift mutated at pNL4-3 so that it does not express the Env protein and the Vpr protein, and into which a luciferase reporter gene has been inserted; all live virus-related assays were performed in a biosafety level 3 facility.