Application of postsynaptic neurotoxin, cardiotoxin, cytotoxin, phospholipase A2 and crude venom of cobra in resisting virus infection
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A cobra family, neurotoxin technology, applied in antiviral agents, medical preparations containing active ingredients, enzymes, etc.
Pending Publication Date: 2020-09-04
沈喆景
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[0009] Although the virus has no other functional structures in the cell except the genetic material, its protein shell is the same as the main component of the cell membrane, and membrane toxins can easily pass through the membrane structure and then destroy internal structures such as mitochondria and lysosomes Traits may also be mechanisms by which snake venom fights viruses
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Embodiment 1
[0151] Example 1. Separation and purification of Chinese cobra crude venom to prepare postsynaptic neurotoxin, cytotoxin, cardiotoxin and phospholipase A2
[0152] Carry out cation exchange by TSK CM-650 (M) column with crude venom of cobra sinensis, the method for separating various toxins comprises the following steps:
[0153] 1. Sample preparation-dissolve 1g of Chinese cobra venom in the ammonium acetate buffer of 25ml 0.025 mole pH6.0, centrifuge at low temperature, and get the supernatant;
[0154] II. Equilibrium - equilibrate the TSK CM-650(M) column with 0.025 mole ammonium acetate solution of pH 6.0;
[0155] III. Elution - After loading the sample, use 0.1-0.5 mole and 0.7-1.0 mole ammonium acetate buffer solution, pH 5.9, to carry out 2-chamber gradient elution, UV detection parameter: 280nm; elution flow rate: 48ml / h;
[0156] IV. Collect various toxin components according to the recorded spectrum, and 12 protein peaks are eluted from the collected solution;
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Embodiment 2
[0159] Example 2. The isolated Chinese cobra post-synaptic neurotoxin, cytotoxin, cardiotoxin, phospholipase A2, and Chinese cobra crude venom were used for antiviral activity experiments, that is, virus plaque reduction rate % experiments
[0160] Principle of viral plaque
[0161] On a single layer of cells covered with a thin layer of agar, after the virus infects a certain cell, due to the limitation of the solid medium, the released virus can only expand from the initially infected cell to the periphery, that is to say, the proliferated virus particles can only spread to neighboring cells. After several cycles of proliferation, a localized area of diseased cells forms, eventually forming a plaque similar to a phage plaque. A plaque is formed by the replicative spread of a virus particle that initially infected a cell and is therefore an accurate measure of virus infectivity.
[0162] Plaque reduction rate% is an indicator of the ability to inhibit virus infection, and...
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Abstract
The invention relates to application of a postsynaptic neurotoxin, a cardiotoxin, a cytotoxin, a phospholipase A2 and a crude venom of a cobra in resisting virus infection. A respiratory tract is easily invaded and infected by viruses due to an open anatomical structure to the outside, and viral influenza and pneumonia are relatively common cases in daily life; and in addition, hepatitis B virusesand AIDS viruses can infect human beings to cause diseases through various ways. The current treatment means of respiratory diseases caused by viruses is not ideal in vaccine or drug effect, and thetreatment effect of viral hepatitis and AIDS needs to be further improved, so that a product which has broad-spectrum inhibition capacity on viruses and has small side effect is developed to become aclinical urgent need. The postsynaptic neurotoxin, the cardiotoxin, the cytotoxin, the phospholipase A2 and the crude venom of the cobra show the effectiveness in resisting virus infection, and possibly become a candidate of a new antiviral product.
Description
Technical field: [0001] The invention relates to the application of main toxin components and crude venom of cobra snakes in treating diseases caused by virus infection, and belongs to the field of biochemistry and biopharmaceuticals. Background technique: [0002] The earth is a space where many organisms coexist. When the environment of this space changes greatly due to natural or man-made reasons, other organisms that originally coexisted peacefully with us may also undergo corresponding changes. Human beings and these organisms The relationship between the virus and the virus will also change accordingly. It is a common example in daily life to infect humans after the virus mutates. [0003] The respiratory tract is vulnerable to virus invasion and infection due to its open anatomical structure to the outside world. Viral influenza and pneumonia are relatively common cases in daily life; in addition, hepatitis B virus and HIV can also infect humans through various channe...
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