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Application of copper gluconate in preparation of a medicine for preventing or treating novel coronavirus infection

A technology of copper gluconate and coronavirus, which can be used in antiviral agents, pharmaceutical formulas, and resistance to vector-borne diseases, etc., and can solve problems such as the lack of vaccines

Inactive Publication Date: 2020-10-02
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The current research on COVID-19 involves many aspects such as epidemiological investigation, clinical research, and virus detection methods, but the research on SARS-Cov-2 virus invasion, replication, and immune regulation mechanism is still in the exploratory stage, and there is no Research on antiviral drugs against SARS-CoV-2 is of great significance as vaccines can provide adequate protection for those who are not sick

Method used

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  • Application of copper gluconate in preparation of a medicine for preventing or treating novel coronavirus infection
  • Application of copper gluconate in preparation of a medicine for preventing or treating novel coronavirus infection
  • Application of copper gluconate in preparation of a medicine for preventing or treating novel coronavirus infection

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Embodiment 1

[0039] 1. Expression and purification of novel coronavirus PLpro protein

[0040] The gene of the new coronavirus PLpro protein is optimized according to the G / C content in its original coding sequence and the application of codons suitable for the prokaryotic expression system of Escherichia coli. The optimized coding nucleotide sequence is shown in SEQ ID NO.1. The translated amino acid sequence is shown in SEQ ID NO.2.

[0041] Choose PET-28 as the expression vector, transform the PET-28 recombinant plasmid with correct sequencing into E.coli BL21(DE3), pick a single clone to expand culture, and culture it on a shaker at 37°C until OD 600 1.0-1.2, add IPTG (final concentration: 0.1mmol / L), induce overnight expression at 20°C; centrifuge the bacterial solution at 4,500rpm at 4°C for 30min, discard the supernatant to collect the bacterial cells. Resuspend the bacteria into a 50ml centrifuge tube with lysate (20mM HEPES, 0.5M NaCl, pH 7.4, add 200μg / ml lysozyme and 0.05% TrRo...

Embodiment 2

[0044] Embodiment 2: Inhibitor copper gluconate half inhibitory concentration IC 50 Determination of

[0045] Copper gluconate was prepared in step concentrations with deionized water.

[0046] The initial rate of the enzymatic reaction of the novel coronavirus PLpro was determined at different concentrations of copper gluconate. The buffer system for the determination of the PLpro activity of the new coronavirus is: 20mmol / L phosphate buffer (pH 6.8). The total concentration of PLpro in the experimental group is 40nmol / L, at the incubation temperature of 25°C, add 200nmol / L, 100nmol / L, 50nmol / L, 25nmol / L, 12.5nmol / L, 6.25nmol / L, 3.13nmol / L , 1.57nmol / L copper gluconate, incubated with shaking at room temperature for 15min, and quickly added fluorescent substrate 1 (Cbz-RLRGG-AMC) or fluorescent substrate 2 (Ub-AMC); in this embodiment, select two concentrations respectively 2.4μmol / L and 7.2μmol / L fluorescent substrate Cbz-RLRGG-AMC and the concentration of 2.5μmol / L fluor...

Embodiment 3

[0049] Embodiment 3: Inhibitor Copper Gluconate Kinetic Determination

[0050] Fluorescent substrate 1 at concentrations of 1.25 μmol / L, 2.5 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L, 40 μmol / L, 60 μmol / L, 80 μmol / L, and 0 nmol / L, 25 nmol / L, Under the concentration of 50nmol / L, 100nmol / L copper gluconate, the PLpro enzyme activity of the new coronavirus was determined. The buffer system for the determination of the PLpro activity of the new coronavirus is: 20mmol / L phosphate buffer (pH 6.8). The total concentration of PLpro in the experimental group was 40nmol / L. At an incubation temperature of 25°C, copper gluconate was added, incubated with shaking at room temperature for 15 minutes, and fluorescent substrate 1 was added quickly, and fluorescence readings were recorded every 1.5 minutes for a total of 30 minutes. The same volume of buffer was added to the control group, and the rest of the experimental conditions were kept the same as the experimental group. The instrument us...

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Abstract

The invention discloses application of copper gluconate in preparation of a medicine for preventing or treating novel coronavirus infection. The molecular formula of the copper gluconate is Cu (C6H11O7) 2, and the copper gluconate can inhibit the activity of novel coronavirus PLpro protein. The inhibition type of the copper gluconate is competitive inhibition, and the inhibition constant Ki valueis 55.3 nmo / L. Tests prove that the copper gluconate can be used for preparing drugs for preventing or treating novel coronavirus infection, and when the copper gluconate is used, the copper gluconatecan be independently used or compounded with drugs, compounds and carriers to form various forms of pharmaceutical compositions such as oral preparations, injections, coatings, lotions, aerosols, oilpreparations or transdermal patches. The invention provides a new compound and a new idea for prevention or treatment of novel coronavirus.

Description

technical field [0001] The invention belongs to the field of medicines, in particular to the application of copper gluconate in the preparation of medicines for preventing or treating novel coronavirus infection. Background technique [0002] Since the first case of pneumonia caused by a new type of coronavirus was discovered in my country in December 2019, the new crown pneumonia has gone through several stages from local outbreaks to rapid expansion and spread. The epidemic has developed rapidly, and confirmed cases have appeared in many countries. On February 11, 2020, the International Virus Taxonomy Committee named the new coronavirus as "severe acute respiratory syndrome coronavirus 2" (sever acute respiratory syndrome coronavirus 2, SARS-Cov-2); WHO named SARS-Cov-2 The disease caused by the infection is named "coronavirus disease-19 (COVID-19)". COVID-19 is transmitted by inhalation or contact with infected droplets, with an incubation period of 2 to 14 days or even...

Claims

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Application Information

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IPC IPC(8): A61K31/30A61P31/14A61K31/191
CPCA61K31/30A61P31/14Y02A50/30
Inventor 柳晓春徐锡明袁文敏王鹏赵晨阳杨金波
Owner OCEAN UNIV OF CHINA
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