Use of periandrin in the preparation of antiviral drugs

Abstract

The application discloses application of dehydropeimine or a pharmaceutical salt thereof in preparation of an antiviral medicine. The small-molecule dehydropeimine hydrochloride screened from a natural product library can significantly inhibit KSHV lytic replication, and the application of dehydropeimine in KSHV antiviral treatment is reported for the first time. The effective drug concentration is low, and the dehydropeimine has high biological safety. The application provides a new solution for KSHV treatment.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to application of dehydropegoine in preparation of an antiviral drug. BACKGROUND

[0002] Herpes virus is a kind of virus with envelope and double-stranded DNA genome. Currently, eight kinds of herpes viruses that can infect human beings have been found, among which KSHV and EBV belong to gamma-herpes virus. After initial infection, herpes virus enters latent infection period, showing lifelong carrying mode. Under certain conditions, the virus is activated to enter lysis replication period, which can cause various diseases such as tumors.

[0003] Kaposi's sarcoma-associated herpesvirus (KSHV) is an important human tumor virus, which is closely related to the occurrence of Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and multicentric Castleman disease (MCD) and other malignant diseases. Kaposi's sarcoma is a common tumor and cause of death in AIDS patients. At present, there is still a lack of efficient antiviral drugs.

[0004] Dehydropegoine, also known as harmine and psilocin, has a molecular formula of C 13 H 12 N2O, CAS: 442-51-3. Literature has reported that its pharmacology mainly has effects on central nervous system, cardiovascular system, muscle and ion channel, such as anti-radiation effect, anti-inflammatory effect, analgesic and antipruritic effect, anti-echinococcus effect, anti-psoriasis effect and the like. SUMMARY

[0005] The present application provides application of dehydropegoine in preparation of an antiviral drug.

[0006] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0007] In a first aspect of the present application, application of dehydropegoine or a pharmaceutically acceptable salt thereof in preparation of an antiviral drug is provided.

[0008] In a preferred embodiment of the present application, it is particularly Kaposi's sarcoma-associated herpesvirus (KSHV).

[0009] The pharmaceutically acceptable salt is an acid addition salt of harmine with hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.

[0010] The application uses harmine or a pharmaceutically acceptable salt thereof as a single active ingredient.

[0011] The second aspect of the application also provides a pharmaceutical preparation made of harmine or a pharmaceutically acceptable salt thereof and a medically acceptable auxiliary or other pharmaceutically acceptable ingredient.

[0012] The dosage form of the pharmaceutical preparation is selected from powders, tablets, granules, capsules, solutions, emulsions and suspensions.

[0013] In a preferred embodiment of the application, the pharmaceutically acceptable salt of harmine used is an acid addition salt of harmine with hydrochloric acid, i.e. harmine hydrochloride (CAS No. 343-27-1 (Liu, H., Han, D., Liu, Y. et al. Harmine hydrochloride inhibits Akt phosphorylation and depletes the pool of cancer stem-like cells of glioblastoma. J Neurooncol 112, 39-48 (2013). https: / / doi.org / 10.1007 / s11060-012-1034-x)), the structure of which is shown below:

[0014]

[0015] Thanks to the above technical solution, the application has the following advantages and beneficial effects:

[0016] The drug small molecule harmine hydrochloride screened from 902 natural product libraries can significantly inhibit the lytic replication of KSHV, and it is the first time to report the application of harmine in KSHV antiviral, and it is found through drug cytotoxicity test that 8 μM harmine treated iSLK219 cells still maintain 100% cell activity; the drug IC 50 required by harmine hydrochloride to inhibit KSHV is only 0.1135 μM; not only is the effective drug concentration low, but also has high biological safety.

[0017] Ganciclovir, a clinically used anti-herpesvirus drug, has an IC50 of 5.1 μΜ against KSHV in the BCBL1 cell model 50 (Medveczky MM, Horvath E, Lund T, Medveczky PG. In vitro antiviral drug sensitivity of the Kaposi's sarcoma-associated herpesvirus. AIDS. 1997 Sep; 11(11): 1327-32. doi: 10.1097 / 00002030-199711000-00006. PMID: 9302441.).

[0018] The present application provides a new solution for the treatment of KSHV. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 Figure 1 is a schematic representation of the results of the growth curve of KSHV in the lytic replication phase in Example 1.

[0020] Figure 2 Figure 2 is a schematic representation of the results of the fitted curve of the inhibition of viral replication by harmine hydrochloride in Example 1. DETAILED DESCRIPTION

[0021] In order to more clearly illustrate the application, the application will be further described below with reference to the preferred embodiments. It should be understood by those skilled in the art that the specific descriptions below are illustrative rather than limiting, and should not limit the scope of protection of the application.

[0022] The acid addition salt of harmine and hydrochloric acid used in the embodiments of the present application is harmine hydrochloride, with a CAS number of 343-27-1.

[0023] Example 1

[0024] Cytotoxicity test of harmine hydrochloride

[0025] iSLK.219 cells constructed by Dr. Don Ganem's Lab were used as a cell model for screening KSHV antiviral drugs (Myoung J, Ganem D. Generation of a doxycycline-inducible KSHV producer cell line of endothelial origin: maintenance of tight latency with efficient reactivation upon induction. J Virol Methods. 2011 Jun;174(1-2):12-21. doi: 10.1016 / j.jviromet.2011.03.012. Epub 2011 Mar 17. PMID: 21419799; PMCID: PMC3095772.). iSLK.219 cells carry a recombinant KSHV.219 virus, and the viral genome contains a constitutive GFP reporter gene and a RFP reporter gene for Doxycycline (Dox) induced RTA expression, which monitor the latent infection activity and the lytic replication activity of the virus, respectively. The addition of Dox and NaB induces iSLK.219 cells to enter the lytic replication state, and the supernatant is collected to detect the viral particle copy number to detect the replication of the virus.

[0026] As shown in Figure 1 , Figure 1 is a schematic diagram of the growth curve results of KSHV in the lytic replication phase in Example 1. As can be seen from the figure, after the induction of Dox and NaB, iSLK.219 cells enter the KSHV virus lytic replication state, and the viral particles in the supernatant are collected and the viral DNA is extracted, and it is found that with the extension of the induction time, the replication of KSHV shows an upward trend.

[0027] DMSO was used to dilute dehydropeimine hydrochloride to prepare 1000x different drug concentration stock solutions. The working concentrations of dehydropeimine hydrochloride were 10 μM, 8 μM, 6 μM, 5 μM, 2 μM, 1 μM, 0.8 μM, 0.6 μM, 0.4 μM, 0.2 μM, 0.1 μM, 0.01 μM, 0.001 μM, and 0 μM (as a control). After drug cytotoxicity test, it was found that compared with the control group, 8 μM dehydropeimine treated iSLK219 cells still maintained 100% cell activity.

[0028] Example 2

[0029] Antiviral activity test of dehydropeimine hydrochloride

[0030] Further investigation was conducted to determine the antiviral activity of dehydrocamellia alkaloid against KSHV. iSLK.219 cells were seeded in 96-well cell culture plates at approximately 4 × 10⁶ cells per well. 4 Cells were cultured for 24 hours until 90% confluence. The culture medium was then replaced with medium containing 1 μg / μL DOX and 1 mM NaB. Simultaneously, 1000× stock solution was added to the medium to dilute to the pre-set working drug concentration. The cells were cultured in a cell culture incubator for another 48-72 hours. The 96-well cell culture plate was removed, the cell supernatant was collected, and viral DNA was extracted for qPCR detection. The inhibition rate of viral replication by dehydrocamellia hydrochloride was calculated and plotted to obtain the drug's IC50 value. 50 It is 0.1135 μM.

[0031] The results are as follows Figure 2 As shown, Figure 2 A schematic diagram showing the fitted curve results for the inhibition of viral replication by Harmine Hydrochloride. From... Figure 2 As can be seen, the inhibitory effect on KSHV virus replication is greater with the increase of the concentration of dehydrocamellia hydrochloride. The drug concentration required to inhibit the virus by 50% is 0.1135 μM.

[0032] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. The application of dehydrocamelin or its pharmaceutical salt in the preparation of antiviral drugs, characterized in that, The virus in question refers to Kaposi's sarcoma-associated herpesvirus.

2. The application according to claim 1, characterized in that, The medicinal salt is an acid addition salt formed by dehydrocamellia alkaloid and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, or mandelic acid.

3. The application according to claim 1 or 2, characterized in that, In this application, dehydrocamelin or its medicinal salt is used as the single active ingredient.

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