阅读说明：本技术 蟾酥提取物在制备治疗SARS-CoV-2病毒引起的感染性疾病的药物中的应用 (Application of venenum Bufonis extract in preparing medicine for treating infectious diseases caused by SARS-CoV-2 virus ) 是由 曹鹏 陈姣 胡星星 于 2020-04-24 设计创作，主要内容包括：本发明公开了蟾酥提取物在制备治疗SARS-CoV-2病毒引起的感染性疾病的药物中的应用,首次发现蟾酥提取物对当前正在全球大流行的SARS-CoV-2具有较好的抑制作用,临床试验表明,蟾酥注射液对COVID-19重症患者的呼吸功能改善作用显著。本发明通过临床试验和体外抗病毒实验证明了蟾酥提取物具有抗新型冠状病毒SARS-CoV-2的作用,对防控和治疗此类病毒引发的感染具有重大意义,尤其为当前正在大流行的SARS-CoV-2引起的COVID-19提供有效的预防和治疗候选药物,也可为未来有可能爆发的新型病毒的防控和治疗提供候选药物。(The invention discloses an application of a toad venom extract in preparing a medicament for treating infectious diseases caused by SARS-CoV-2 virus, wherein the toad venom extract is found to have a better inhibiting effect on SARS-CoV-2 which is currently in global pandemic for the first time, and clinical tests show that the toad venom injection has a remarkable effect on improving the respiratory function of a COVID-19 severe patient. Clinical tests and in vitro antiviral experiments prove that the toad venom extract has the function of resisting novel coronavirus SARS-CoV-2, has great significance for preventing, controlling and treating the infection caused by the viruses, particularly provides effective prevention and treatment candidate medicines for the COVID-19 caused by the SARS-CoV-2 which is currently in a pandemic, and also provides candidate medicines for the prevention, control and treatment of novel viruses which are likely to break out in the future.)

1. Application of Bufonis venenum extract in preparing medicine for treating infectious diseases caused by SARS-CoV-2 virus is provided.

2. The use of claim 1, wherein the infectious disease caused by SARS-CoV-2 virus comprises COVID-19, upper respiratory infection or gastroenteritis.

3. The use of claim 1, wherein the toad venom extract is prepared by:

(1) taking dried toad venom, cleaning, soaking in 8-10 times of injection water by mass for 0.5-24 h, grinding, adding ethanol to enable the ethanol content in an extracting solution to reach 50-90 vt%, and then standing and extracting at 10-30 ℃ for 1-6 days;

(2) filtering the extraction solution obtained in the step (1), concentrating the filtrate until no alcohol smell exists, adding water for injection, stirring uniformly, refrigerating and standing at a low temperature of 0-5 ℃ for 30-60 hours, filtering to separate the filtrate, and concentrating to obtain the compound.

4. The use of claim 1 or 3, wherein the effective component of the Bufonis venenum extract comprises at least one monomer selected from bufalin, cinobufagin, and bufogenin, or a combination of two or more monomers.

5. The use as claimed in claim 4, wherein the effective component of the Bufonis venenum extract is a composition of bufalin, cinobufagin and resibufogenin, and the total concentration of the three in the Bufonis venenum extract is 10-53 μ g/m L.

6. The use of claim 4, wherein the mass ratio of the three monomers of bufalin, cinobufagin and bufogenin is (4.5-6): (2.5-4).

7. The use of claim 3, wherein the Bufonis venenum extract can be combined with pharmaceutically acceptable carrier to make into liquid injection, powder for injection, tablet, capsule, soft capsule, powder, pill, oral liquid, unguent, nanometer preparation, microsphere preparation, gel preparation, granule or dressing.

Technical Field

The invention belongs to the field of biological medicine, relates to a toad venom and new application of an extract thereof, and particularly relates to application of a toad venom extract in preparation of a medicine for treating infectious diseases caused by SARS-CoV-2 virus.

Background

The hosts of highly pathogenic viruses are mainly wild animals, and with the increasing overlap of human and wild animal ecosystems, the potential for new viruses to appear in humans is also increasing. The periodic circulation of viruses as xenogeneic members into and out of human and zoonotic hosts has made the development of specific antiviral therapies and vaccines exceptionally complex. The anti-coronavirus medicine with definite curative effect and broad spectrum is searched, which not only can relieve the urgent need of spreading SARS-CoV-2, but also provides candidate medicines for preventing and treating novel viruses which are likely to be outbreak in the future.

The traditional Chinese medicine Bufonis venenum is the dry secretion of skin gland and retroauricular gland of Bufo bufo gargarizans Cantor or Bufo melanostictus Schneider of Bufo siccus. The Bufonis venenum mainly contains bufadienolides, non-cardiotonic sterols, indole derivatives and other chemical components, and has pharmacological activities of tonifying heart, increasing blood pressure, relieving pain due to anesthesia, resisting bacteria, enhancing immunity, and resisting tumor. The venenum bufonis Chinese medicinal preparation is widely applied to clinical practice of anti-tumor treatment, and has a certain treatment effect on lung cancer, leukemia, liver cancer, gastric cancer, gynecological malignant tumors and the like. The chemical composition of toad venom is complex, and mainly comprises bufadienolides, 20, 21-epoxy bufadienolides, cardiotonic steroid alkene toad venom, indole derivatives, toad venom methyl alkali, sterols, peptides, amino acids and the like. Bufalin, bufogenin and cinobufagin are 3 main active monomers of venenum bufonis.

Disclosure of Invention

The invention aims to solve the problem of clinical lack of effective medicaments for treating COVID-19 and provide a medicament for safely and effectively inhibiting SARS-CoV-2 virus.

The technical scheme is as follows: the invention discovers that the toad venom extract can be used as an inhibitor of SARS-CoV-2 virus for the first time, inhibits the virus from infecting host cells, and verifies the treatment effect of the toad venom extract on COVID-19 through clinical tests. Thereby providing the application of the toad venom extract in preparing the medicine for treating infectious diseases caused by SARS-CoV-2 virus.

Specifically, the infectious diseases caused by SARS-CoV-2 virus include, but are not limited to, COVID-19, upper respiratory tract infection, gastroenteritis, and the like.

Specifically, the toad venom extract is prepared by the following steps:

(1) taking dried toad venom, cleaning, soaking in 8-10 times of injection water by mass for 0.5-24 h, grinding, adding ethanol to enable the ethanol content in an extracting solution to reach 50-90 vt%, and then standing and extracting at 10-30 ℃ for 1-6 days;

(2) filtering the extraction solution obtained in the step (1), concentrating the filtrate until no alcohol smell exists, adding water for injection, stirring uniformly, refrigerating and standing at a low temperature of 0-5 ℃ for 30-60 hours, filtering to separate the filtrate, and concentrating to obtain the compound.

Specifically, the effective components of Bufonis venenum extract at least include bufalin (formula C)₂₄H₃₄O₄) Cinobufagin (Cinobufagin, molecular formula C)₂₆H₃₄O₆) Ester bufogenin (Resibufogenin, molecular formula C)₂₄H₃₂O₄) Any one monomer or a combination of two or more monomers.

Preferably, the active ingredients in the toad venom extract are a composition of bufalin, cinobufagin and resibufogenin, and the total concentration of the three in the toad venom extract is 10-53 mu g/m L.

Most preferably, the mass ratio of three monomers of bufalin, cinobufagin and bufogenin which are effective components in the toad venom extract is (4.5-6): (2.5-4): (2.5-4).

The invention adopts dried toad venom product to extract toad venom extract, and then can prepare liquid injection, powder injection, tablets, capsules, soft capsules, powder, pills, oral liquid, paste, nano preparations, microsphere preparations, gel preparations, granules or dressings and other formulations by matching with pharmaceutically acceptable carriers.

Has the advantages that:

(1) the invention discovers that the toad venom extract has a good inhibition effect on SARS-CoV-2 which is currently in global pandemic for the first time, and clinical tests show that the toad venom injection has a remarkable effect on improving the respiratory function of the COVID-19 severe patients, while clinical experience proves that lopinavir and ritonavir which are medicines have an unobvious effect on the COVID-19 severe patients;

(2) clinical tests and in vitro antiviral experiments prove that the toad venom extract has the function of resisting novel coronavirus SARS-CoV-2, has great significance for preventing, controlling and treating the infection caused by the viruses, particularly provides effective prevention and treatment candidate medicines for the COVID-19 caused by the SARS-CoV-2 which is currently in a pandemic, and also provides candidate medicines for the prevention, control and treatment of novel viruses which are likely to break out in the future.

Drawings

The foregoing and/or other advantages of the invention will become further apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings.

FIG. 1 is the fingerprint of the toad venom injection prepared by the invention; wherein, RT57.7 peak is bufalin; RT67.9 peak is cinobufagin; the RT69.5 peak is resibufogenin.

Detailed Description

The invention will be better understood from the following examples.