Application of Ventolin in the preparation of drugs against respiratory syncytial virus

Abstract

The invention provides an application of vindoline in preparing an anti-respiratory syncytial virus drug, belonging to the technical field of traditional Chinese medicine. Experiments have shown that vindoline has an obvious inhibitory effect on respiratory syncytial virus A2 beads and B beads. Therefore, using it as an active ingredient in preparing an anti-respiratory syncytial virus drug has important application value.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicines, and in particular to an application of vindoline in the preparation of an anti-respiratory syncytial virus drug. Background Art

[0002] The medicinal plant Catharanthus roseus is a herbaceous plant of the Apocynaceae family. It mainly grows in tropical and subtropical areas. It is widely grown in Guangdong and Guangxi in my country. It is also cultivated in southern Jiangsu and has been introduced in Wuhan. Vindoline (also known as Wenduoling / Lin, Changchun Flower Ning, Changchun Daoling) is a substance extracted from the leaves of Catharanthus roseus. It is a precursor for the biosynthesis and production of important anticancer drugs vincristine and vinblastine. Vindoline has been shown to have anti-tumor, anti-diabetic, anti-apoptosis, antioxidant and anti-inflammatory effects.

[0003] Human respiratory syncytial virus (RSV) belongs to the family of Paraadhesinviridae. It is a negative-sense single-stranded RNA virus that causes epidemic respiratory infections, usually with seasonal outbreaks, peaking in winter in temperate climates and rainy seasons in tropical climates. It is divided into RSV-A and RSV-B types based on the structure of the surface protein. It is a cause of illness and death in children, the elderly, and immunocompromised patients. In addition to general symptomatic treatments such as antiasthmatic, expectorant, and anti-inflammatory, the current recommended treatment for RSV is the broad-spectrum antiviral drugs ribavirin and palivizumab, but due to their high cost, they cannot be widely used, so there is a need to find natural medicines to solve this problem. Summary of the invention

[0004] The main purpose of the present invention is to provide a use of vindoline in the preparation of an anti-respiratory syncytial virus drug in response to the defects of the prior art; the vindoline has a good anti-respiratory syncytial virus effect.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0006] Wenduling is used in the preparation of anti-respiratory syncytial virus drugs.

[0007] Preferably, the concentration of vindoline in the anti-respiratory syncytial virus drug is ≥ 0.01 wt %.

[0008] Compared with the prior art, the present invention has the following beneficial effects:

[0009] Vindoline comes from the natural medicinal plant Catharanthus roseus, which is easy to obtain, low-cost, and has few side effects. It has a strong ability to resist respiratory syncytial virus (A2 beads and B beads). BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The present invention will be further described below in conjunction with the accompanying drawings and embodiments:

[0011] Figure 1 This is a graph showing the inhibitory effects of different concentrations of Vindoline on RSV virus on the sixth day in the examples of the present invention. DETAILED DESCRIPTION

[0012] To avoid redundancy, the items used in the following examples are all commercially available products unless otherwise specified, and the methods used are all conventional methods unless otherwise specified.

[0013] The sources of some raw materials used in the present invention are as follows:

[0014] Ventolin was provided by Wuhan Botanical Garden, Chinese Academy of Sciences;

[0015] Respiratory syncytial virus RSV-A2 strain was provided by Guangzhou National Laboratory;

[0016] The RSV-B strain was provided by the Wuhan Institute of Virology, Chinese Academy of Sciences.

[0017] Example 1

[0018] S1: Cell culture

[0019] Human laryngeal cancer epithelial cells Hep-2 were seeded in 96-well plates containing 10% FBS DMEM medium, with 1x10 4 Cells were cultured overnight in a 37°C, 5% CO2 incubator;

[0020] S2: Prepare the Vindoline dilution solution and the Vindoline virus mixture

[0021] Vindoline was serially diluted with DMEM solution containing 2% FBS to 500 μM, 100 μM, and 20 μM vindoline dilutions, and added to a 96-well plate (50 μl / well). Then, 50 μl of 2×10 3 RSV-A2 strain (infect Hep-2 cells at an infection multiplicity of 0.1) was added and gently pipetted to mix, so that the final concentration of the drug was: 250 μM, 50 μM, and 10 μM; at the same time, 4 groups of control wells were prepared with equal volumes, including a virus-only group (final concentration of 1×10 3 PFU RSV strain), 1% DMSO only group, DMSO + virus group, and 2% DMEM blank group (cell control group), each group was repeated 3 times;

[0022] S3: Hep-2 cells infected with different concentrations of Vindoline

[0023] Use a pipette to discard the culture medium in the 96-well plate containing Hep-2 cells, add the mixture of vindoline and virus prepared in step S2, and culture in a 35°C, 5% CO2 incubator for 6 days;

[0024] S4: Antiviral testing

[0025] The cytotoxicity detection kit (cck8 kit) was used to determine the neutralization ability of different concentrations of vindoline against viruses. The more cells survive, the stronger the neutralization ability of the Chinese medicine at this concentration against RSV. The specific operation is as follows: prepare the cck8 detection solution according to the instructions, that is, add 10μl / well of sterile PBS solution to 100μl / well of cck8 solution, blow and mix well for use. Use a pipette to aspirate the vindoline and virus mixture in the 96-well plate in S3, and add 100μl of cck8 detection solution to each well in the plate. The culture plate was placed in a 35°C, 5% CO2 incubator for 1.5 hours. After the culture plate was placed in an ELISA reader and shaken slowly for 1 minute, the optical density (OD value) at 450nm was measured.

[0026] Example 2

[0027] The operation steps are the same as those in Example 1, except that the strain used is RSV-B strain.

[0028] Virus inhibition rate (%) = [OD450 (different concentrations of drug experimental groups) - OD450 (virus group)] / [OD450 (cell control group) - OD450 (virus group)] * 100%.

[0029] The experimental results are as follows Figure 1 As shown, for RSV-A2 strain, compared with the control group without drug (Control-A2), Wenduling can effectively inhibit the activity of the virus in a concentration-dependent manner (IC50=39.3μM); for RSV-B strain, compared with the control group without drug (Control-B), although Wenduling at concentrations of 10μM and 50μM has certain antiviral ability, the antiviral ability at a concentration of 250μM is more significant (IC50=261.1μM). It can be seen that Wenduling has a significant antiviral effect on respiratory syncytial virus RSV, and can be used to prepare respiratory syncytial virus RSV drugs, providing more options for the clinical treatment of RSV.

[0030] The above embodiments are only preferred technical solutions of the present invention and should not be regarded as limiting the present invention. The protection scope of the present invention shall be the technical solutions recorded in the claims, including equivalent replacement solutions of the technical features in the technical solutions recorded in the claims. That is, equivalent replacement improvements within this scope are also within the protection scope of the present invention.

Claims

1. Application of Ventolin in the preparation of anti-respiratory syncytial virus drugs.

2. The use according to claim 1, characterized in that: The concentration of vindoline in the anti-respiratory syncytial virus drug is ≥0.01wt%.

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