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Application of pyroglutamic acid in the preparation of drugs for the prevention and treatment of novel coronavirus pneumonia

A technology of pyroglutamic acid and coronavirus, applied in the direction of antiviral agents, drug combinations, pharmaceutical formulas, etc., can solve the problems of poor curative effect, achieve low effective concentration, high therapeutic index, and increase the effect

Active Publication Date: 2021-04-23
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the current treatment methods are symptomatic treatment, especially for some severe patients with poor curative effect

Method used

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  • Application of pyroglutamic acid in the preparation of drugs for the prevention and treatment of novel coronavirus pneumonia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Virus amplification

[0027] African green monkey kidney cells (VeroE6) were divided into 3×10 5 Each well was inoculated into a 96-well plate in Eagle’s minimum basal medium (minimum Eagle’s medium, MEM; GibcoInvitrogen) containing 10% fetal bovine serum (FBS; GibcoInvitrogen) at 37°C, 5% CO 2 Cultivate until the monolayer grows. Inoculate a 100-fold dilution of the clinical isolate of the novel coronavirus pneumonia into a 96-well plate full of monolayer cells, place at 37°C, 5% CO 2 Culture for two days (including the normal control group).

[0028] Two days later, the lesion degree reached more than 75%. Put it in a -80°C ultra-low temperature refrigerator, freeze and thaw once repeatedly, collect the virus liquid amplified by the cells, centrifuge at 3000r / min for 30 minutes, remove the sediment, put it in a small tube and put it in a -80°C ultra-low temperature freezer Long-term preservation.

Embodiment 2

[0029] Example 2 L-pyroglutamic acid drug toxicity evaluation

[0030] After dissolving L-pyroglutamic acid powder with DMSO, add culture medium to dilute to 20mg / mL, the final concentration of DMSO is 1%, filter through a 0.22μm filter membrane and store at 4°C; after filtering, store at 4°C. Press about 2.5×10 per hole 4 Cells were seeded into a 96-well plate, and after 24-48 hours, when the cells had grown into a single layer, the culture medium was discarded, and 100 μL / well of drugs of different dilutions were added, and 100 μL / well of MEM was added to the control wells of normal cells, and kept at 37°C in 5% CO 2 Continue culturing for 2 to 5 days, add 20 μL of CCK8 solution (5 mg / mL) to each well, and place at 37°C in 5% CO 2 Continue to incubate for 4 hours in the incubator. Discard the culture supernatant, add 100 μL dimethyl sulfoxide (DMSO) to each well, and shake at low speed for 10 minutes to fully dissolve the crystals. Select a wavelength of 490nm, and measur...

Embodiment 3

[0031] Example 3 Efficacy Evaluation of L-pyroglutamic Acid Against New Coronary Pneumonia Novel Coronavirus

[0032] To evaluate the antiviral efficacy of the drug, VeroE6 cells were grown at a density of 5×10 4Cells / well were cultured overnight in a 48-well cell culture dish. Add virus (MOI 0.05) to infect for 2 hours. Then add 2-fold serially diluted drugs, set up 4 replicate wells for each concentration, take the maximum non-toxic concentration as the initial concentration of the drug, and incubate at 34°C, 5% CO 2 Incubate for 2 days in the incubator. Cytopathogenic Effect (CPE) was recorded. The occurrence of CPE in cells is recorded according to the 6-level standard. After recording the CPE, stain with CCK8 and measure the OD value. Efficacy evaluation of L-pyroglutamic acid against novel coronavirus pneumonia The efficacy of L-pyroglutamic acid was evaluated by the CCK8 method, and the half effective concentration (EC) against new coronary pneumonia novel coronavi...

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Abstract

The invention relates to the application of pyroglutamic acid in the preparation of drugs for the prevention and treatment of novel coronavirus pneumonia, specifically disclosed. Pyroglutamic acid or its pharmaceutically acceptable salts, isotopes, stereoisomers, mixtures of stereoisomers, tautomers, esters, amides or prodrugs are used in the preparation of prevention and / or treatment of diseases caused by coronaviruses application in medicines. Described coronavirus is novel coronavirus SARS-Cov-2, SARS-CoV, HCoV 229E, NL63, OC43, HKU1 and MERS-CoV. The half effective concentration of pyroglutamic acid against the novel coronavirus of new coronary pneumonia is 9.34μM, and the half effective concentration is low, indicating that the antiviral effect is good.

Description

technical field [0001] The invention belongs to the field of antiviral drugs, and in particular relates to the application of pyroglutamic acid in the preparation of drugs for preventing and treating novel coronaviruses against new coronary pneumonia. Background technique [0002] Novel coronavirus pneumonia (Corona Virus Disease 2019) is an infectious disease caused by a new type of coronavirus (SARS-Cov-2) infecting the human body. , 2020, 395(10223):507-513; The Lancet, 2020, 395(10223):497-506]. Coronaviruses belong to the genus Coronavirus in the family Coronaviridae. Viruses of the genus Coronavirus are positive-strand single-stranded RNA viruses with an envelope (envelope), with a diameter of about 80-120nm. Its genetic material is the largest among all RNA viruses, and generally only infects humans, mice, pigs, cats, and dogs. , Avian vertebrates. Coronaviruses were first isolated from chickens in 1937. The shape of coronavirus particles is irregular, with a diam...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/4015A61K45/06A61P31/14A61P11/00
CPCA61K31/4015A61K45/06A61P11/00A61P31/14
Inventor 袁曙光崔文强罗木鹏邹荣峰陈显翀赵金存孙静
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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