Application of vitamin D2 in preparation of SARS-CoV-2 novel coronavirus inhibitor
阅读说明：本技术 维生素D2在制备SARS-CoV-2新型冠状病毒抑制剂中的应用 (Application of vitamin D2 in preparation of SARS-CoV-2 novel coronavirus inhibitor ) 是由 宋世东 孙媛媛 葛以跃 迟莹 江经纬 于 2020-05-20 设计创作，主要内容包括：本发明属于医药技术领域,具体涉及维生素D2在制备SARS-CoV-2新型冠状病毒抑制剂中的应用。本发明通过研究发现,维生素D2可以结合SARS-CoV-2新型冠状病毒的刺突蛋白S1上与ACE-2受体结合的RBD区,阻断病毒进入细胞,从而抑制病毒复制。因此,维生素D2可以作为SARS-CoV-2新型冠状病毒的抑制剂,为预防和/或治疗SARS-CoV-2新型冠状病毒提供了新思路,为临床试验的开展提供了理论基础,具有重要的医学研究价值。(The invention belongs to the technical field of medicines, and particularly relates to application of vitamin D2 in preparation of a novel SARS-CoV-2 coronavirus inhibitor. The invention discovers that vitamin D2 can be combined with RBD area combined with ACE-2 receptor on spike protein S1 of SARS-CoV-2 novel coronavirus to block virus from entering cells, thereby inhibiting virus replication. Therefore, the vitamin D2 can be used as an inhibitor of SARS-CoV-2 novel coronavirus, provides a new thought for preventing and/or treating SARS-CoV-2 novel coronavirus, provides a theoretical basis for the development of clinical tests, and has important medical research value.)
1. Application of vitamin D2 in preparing SARS-CoV-2 coronavirus inhibitor is provided.
2. Use of vitamin D2 in preparation of preparation for inhibiting SARS-CoV-2 novel coronavirus nucleic acid proliferation is provided.
3. Use of vitamin D2 in preparation of preparation for blocking SARS-CoV-2 coronavirus from entering cell.
4. Application of vitamin D2 in preparing medicine for preventing and/or treating SARS-CoV-2 coronavirus pneumonia infection is provided.
5. The use according to any one of claims 1 to 4, wherein vitamin D2 inhibits the recognition of the receptor ACE-2 by the spike protein S1 of the SARS-CoV-2 novel coronavirus.
6. A medicament for preventing and/or treating SARS-CoV-2 novel coronavirus pneumonia infection, which comprises vitamin D2.
7. The medicament of claim 6, wherein the blood concentration of vitamin D2 is 5-20 μmol/L.
8. The medicament of claim 7, wherein the blood concentration of vitamin D2 is 10 μmol/L.
Technical Field
The invention relates to the technical field of medicines, in particular to application of vitamin D2 in preparation of a novel SARS-CoV-2 coronavirus inhibitor.
Background
Highly pathogenic coronavirus infection has become a public health concern over the last decade. Such as severe acute respiratory syndrome (SARS, 2002-2004), middle east respiratory syndrome (MERS, 2012-to-date), SARS-CoV-2 (COVID-19), each disease has a tremendous impact on human health and economic development. SARS-CoV-2 is reported to be a new member of the genus Beta coronavirus (Betacononavirus, Beta-CoV), and is closely related to Severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV) and several bat coronaviruses.
Because there are no effective drugs and vaccines, the research and development of new SARS-CoV-2 coronavirus medicines is of great significance, and in the war without smoke, many scientific research institutions and pharmaceutical companies are dedicated to research and develop new coronavirus vaccines and therapeutic medicines. Therefore, the development of a novel therapeutic drug for SARS-CoV-2 coronavirus has great social and economic significance.
Vitamin D2(Vitamin D2; Ergocalceiferol) is a Vitamin D analog, synthesized by certain plants in the presence of UVB light. The medicine is used for treating chondropathy, and the FDA in the United states is approved to be on the market in 1941. According to the us 2011 diagnostic guidelines, ergocaliferol may be used as a dietary supplement to supplement vitamin D, and is as effective as cholecalciferol produced by the skin under uv exposure (vitamin D3). Frank et al (2020) found that in immune responses, the vitamin D-dependent antibacterial pathway is responsive to double-stranded RNA generated during replication of the novel coronavirus COVID-19 and the like. These pathways subsequently up-regulate the clearance of damaged cells (autophagy), as well as various antibacterial and antiviral peptide levels. Thus, vitamin D deficiency may predispose the host to infection with new coronaviruses by preventing the host from activating the above-mentioned defense pathways and affecting the migration of lymphocytes and macrophages. However, at present, no report on whether vitamin D2 can inhibit SARS-CoV-2 is found, and the action mechanism is not clear.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides the application of vitamin D2 in preparing a novel SARS-CoV-2 coronavirus inhibitor. Vitamin D2 can bind to RBD region of spike protein S1 of SARS-CoV-2 coronavirus, which is bound with ACE-2 receptor, and block virus from entering cell, thereby inhibiting virus replication.
Specifically, the invention provides the application of vitamin D2 in preparing a preparation for inhibiting the nucleic acid proliferation of SARS-CoV-2 novel coronavirus.
Specifically, the invention provides the application of vitamin D2 in preparing a preparation for blocking SARS-CoV-2 novel coronavirus from entering cells.
Specifically, the invention provides application of vitamin D2 in preparing a medicament for preventing and/or treating SARS-CoV-2 novel coronavirus pneumonia infection.
Preferably, the vitamin D2 inhibits the recognition of the receptor ACE-2 by the spike protein S1 of the SARS-CoV-2 novel coronavirus.
Another object of the present invention is to provide a pharmaceutical composition for preventing and/or treating SARS-CoV-2 coronavirus pneumonia infection, which comprises vitamin D2.
Preferably, the blood concentration of the vitamin D2 is 5-20 mu mol/L.
More preferably, the blood concentration of the vitamin D2 is 10 mu mol/L.
The receptor ACE-2 is recognized by the spike protein spike (S1) due to the entry of the novel SARS-CoV-2 coronavirus into the cell. Therefore, small molecule inhibitors that inhibit spike interaction with ACE-2 targets may be useful in the prevention and/or treatment of novel SARS-CoV-2 coronavirus. The inventor carries out virtual molecular docking on an ACE-2 receptor binding region on an S protein and scores by an AI scoring system by all FDA approved old drugs (2514, a drug bank database) to obtain 19 candidate old drugs, tests and screens various old drugs, and finds that vitamin D2 can be used as an effective 2019 novel coronavirus inhibitor.
The structural formula of vitamin D2 is shown in figure 1. Chinese name: ergocalciferol (vitamin D2). English name: ergocalciferol (Vitamin D2). The molecular formula is as follows: c28H44And O. Molecular weight: 396.6484. CAS number: 50-14-6.
The inventor finds that Ergocalceiferol can be combined with RBD (ribose-binding domain) which is combined with an ACE-2 receptor on a spike protein S1 of SARS-CoV-2 novel coronavirus, blocks the virus from entering cells, and is a brand-new mechanism inhibitor of SARS-CoV-2 novel coronavirus. Ergocalceiferol inhibits viral nucleic acid proliferation by 57.53% at a cellular level of 10. mu.M.
Compared with the prior art, the invention has the following beneficial effects:
the invention discovers that the vitamin D2 can be used as an inhibitor of SARS-CoV-2 novel coronavirus, and provides a new idea for preventing and/or treating SARS-CoV-2 novel coronavirus; the vitamin D2 is used as an old medicine for new use, has no safety problem, provides a theoretical basis for the development of clinical tests, and has important medical research value.
Drawings
FIG. 1 shows the structure of Ergocalceiferol.
FIG. 2 shows the results of affinity determination of Ergocalceiferol with the RBD region of SARS-CoV-2 novel coronavirus S protein.
Detailed Description
It should be noted that the raw materials used in the present invention are all common commercial products, and the sources thereof are not particularly limited. The following raw material sources are exemplary.
First, experiment reagent and consumable
Recombinant 2019nCOVSpike RBD protein (KACTUS, Cat. COV-VM5 BD);
protein labeling kit (Cat # MO-L011);
capillary loading (Cat # MO-K022);
DMSO (Biyuntian biotechnology, Cat number ST-1276-;
DMEM medium (Gibco, cat # 12430-;
CCK-8 kit (Biyuntian biotechnology, cat # C0039);
nucleic acid extraction kit (Roche, Cat. 03038505001);
2019 novel coronavirus nucleic acid detection kit (Shanghai Berger, Cat. SJ-HX-226-2).
Second, main instrument
QuantStudioDx fluorescent quantitative PCR instrument, manufacturer ABI, No. 04-620;
MagNA Pure LC2.0 full-automatic nucleic acid extractor, manufacturer Roche;
carbon dioxide incubator (GALAXYS), manufacturer RsBiotech, serial number 04-0303;
SARS-CoV-2 virus strain
The BSL-3 laboratory of the Jiangsu provincial disease prevention and control center separates and identifies a novel coronavirus strain through a separation culture laboratory. The strain numbers used in this experiment are: BetacoV/JS03/human/2020(EPI _ ISL _ 411953).
Tetra, compound Ergocalceiferol
IUPAC name: (1S,3Z) -4-methylene-3- [ (2E) -2- [ (1R,3aS,7aR) -octahydro-7a-methyl-1- [ chemical book [1R,2E,4R) -1,4, 5-trimethyl-2-hexen-1-yl]-4H-indan-4-propylene]Ethylene radical]Cyclohexanol (1S,3Z) -4-methyl-3- [ (2E) -2- [ (1R,3aS,7aR) -octahydro-7a-methyl-1- [ (1R,2E,4R) -1,4, 5-trimethyl-2-hexen-1-yl]-4H-inden-4-ylidene]ethylidene]Cyclohexanol, molecular weight 396.65, formula C28H44O, purity: 99.89%, storage conditions: -20 ℃ (powder); -80 ℃ (solvent). The goods number is: t1086. The compound samples and related information were provided by Shanghai ceramic Biotechnology, Inc.