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Application of cyclic adenylic acid and its derivatives in anti-ev71 virus

A technology of cyclic AMP and cyclic AMP calcium, which is applied in the field of application of cyclic AMP and its derivatives in anti-EV71 virus, and can solve the problems that the inhibitory activity has not been reported.

Active Publication Date: 2021-03-16
WUHAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antiviral activity of cAMP, including the inhibitory activity against EV71 virus, has not been reported so far.

Method used

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  • Application of cyclic adenylic acid and its derivatives in anti-ev71 virus
  • Application of cyclic adenylic acid and its derivatives in anti-ev71 virus
  • Application of cyclic adenylic acid and its derivatives in anti-ev71 virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Cytotoxicity detection of embodiment 1 cyclic AMP (cAMP)

[0027] In RD cells, the cytotoxicity of cyclic AMP (cAMP) was tested. RD cells were seeded in 96-well plates, cultured in a 5% CO2 incubator at 37°C for 12-16 hours, discarded the cell culture medium, and added cell maintenance solutions containing different concentrations of test cyclic AMP to continue the culture. Group 3 duplicate wells, the control group added the same amount of PBS. After 48 hours of treatment, stain with MTT, detect OD492nm, and analyze cell viability.

[0028] Result analysis: The test results are as follows: figure 1 As shown, the Prism7 software calculates the median cyctoxic concentration (CC50) of the drug on the cells, and the CC50 of cyclic AMP (cAMP) is 50.59 mM. In subsequent implementation cases, the maximum concentration of cAMP used is 5mM, which is within the safe and non-toxic range.

Embodiment 2

[0029] Embodiment 2 Cyclic AMP (cAMP) detects the antiviral activity of EV71

[0030] Plate the RD cells on a 96-well cell culture plate, culture them in a 5% CO2 incubator at 37°C for 12-16 hours, infect the cells with 100 TCID50 of the EV71 virus solution for 2 hours, and add cell maintenance solutions containing different concentrations of test cyclic adenosine monophosphate to continue After culturing for about 48 hours, add the same volume of PBS to the negative control. When about 90% of the CPE lesions appeared in the virus control wells, the cytopathic effect (CPE) was observed under a microscope.

[0031] The inhibitory rate of drugs on EV71 was detected by MTT method. The specific steps are: add 50 μL (5 mg·mL-1) of MTT to each well, remove the supernatant after incubation for 4-6 hours, and add an equal volume of DMSO to dissolve the precipitate. Read the corresponding absorbance (OD492 value) at 492 nm with a microplate reader.

[0032] Result analysis: cyclic A...

Embodiment 3

[0033] Example 3 Detection of Cyclic Adenosine Monophosphate (cAMP) Inhibiting EV71 Proliferation Activity

[0034] RD cells were seeded in 24-well cell culture plates, cultured in a 5% CO2 incubator at 37°C for 12-16 hours, infected with 100 TCID50 of EV71 virus liquid for 2 hours, and maintained with cells containing different concentrations of experimental cyclic adenosine monophosphate The same volume of PBS was added to the negative control. After 24 hours of treatment, the effect of inhibiting virus proliferation was detected by titer.

[0035] Result analysis: The test results are as follows: Figure 4 As shown, under different concentration conditions, the compound cyclic AMP (cAMP) has a significant inhibitory effect on EV71.

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Abstract

The invention discloses application of cyclic adenosine monophosphate and a derivative thereof in resistance of EV71 virus. The derivative comprises 8 bromine-cyclic adenosine monophosphate and dibutyl acyl-cyclic adenosine monophosphate calcium salt; the cyclic adenosine monophosphate and the derivative thereof are used as EV71 virus resistant drugs; when the concentration of the cyclic adenosinemonophosphate and the dibutyl acyl-cyclic adenosine monophosphate calcium salt is 800 mug / mL, the EV71 virus inhibition rate is 99.8% and 92.8% respectively; and, when the concentration of the 8 bromine-cyclic adenosine monophosphate is 500 mug / mL, the EV71 virus inhibition rate is 98%. New use of the cyclic adenosine monophosphate and the derivative thereof used for treating EV71 infectious diseases is provided in the invention at the first time; the cyclic adenosine monophosphate has a certain guidance role on development of the EV71 virus resistant activity; reference is provide for further developing an EV71 virus resistant drug; furthermore, the cyclic adenosine monophosphate, the 8 bromine-cyclic adenosine monophosphate and the dibutyl acyl-cyclic adenosine monophosphate calcium salt are simple in synthesis process; and thus, large-scale production and popularization are easily carried out.

Description

technical field [0001] The invention belongs to the technical field of antiviral drugs, and specifically relates to the application of cyclic adenosine acid and derivatives thereof in anti-EV71 virus. Background technique [0002] Enterovirus 71 (EV71), a member of the genus Enterovirus in the family Picornaviridae, is one of the most important pathogens causing hand, foot and mouth disease in infants, sometimes accompanied by severe central nervous system complications. Including aseptic meningitis, brainstem encephalitis, autonomic nervous disorder, pulmonary edema, etc., and even death. Since it was first reported in 1969, EV71 infectious disease has broken out and spread many times around the world, especially in the Asia-Pacific region, especially in my country. At present, the prevention and treatment of viral diseases mainly rely on vaccines and drugs. The relevant vaccines were only launched in 2015. There is no specific data to support that the marketed vaccines ca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7076A61P31/14
CPCA61K31/7076A61P31/14
Inventor 吴莹杨庆雨
Owner WUHAN UNIV
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