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Pharmaceutical composition for preventing and/or treating coronavirus infection as well as preparation method and application thereof

A composition and drug technology, applied in the field of medicine, can solve problems such as undeveloped therapeutic drugs, achieve the effect of increasing the number and efficiency of action, strong specificity, and not easy to reject reactions

Active Publication Date: 2020-11-24
海图生物科技(上海)有限责任公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Effective therapeutic drugs against SARS CoV-2 infection have not yet been developed

Method used

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  • Pharmaceutical composition for preventing and/or treating coronavirus infection as well as preparation method and application thereof
  • Pharmaceutical composition for preventing and/or treating coronavirus infection as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Preparation of Antigenic Peptides

[0063] Using software CLC Protein Workbench version 5.8 and Signal P5.0 software, SYFPE1TH1 remote prediction database, PREDEPP database to screen potential T cell single epitopes in SARS CoV-2 Env protein and cell membrane glycoproteins, and obtain potential HLA-A2 restricted Three single-epitope T cell antigen peptides: FLLVTLAIL, FLFLTWICL, SMWSFNPET.

[0064] The single-epitope T cell antigen peptide and the dendritic pluripotent T cell epitope peptide can be synthesized by conventional solid-phase synthesis. The amino acid sequence of each epitope peptide can also be synthesized by a peptide synthesis company.

Embodiment 2

[0065] Example 2 Preparation of dendritic cells loaded with pluripotent T cell epitope peptide

[0066] A method for preparing dendritic cells loaded with pluripotent T cell epitope peptides, comprising the steps of:

[0067] Human peripheral blood PBMCs were labeled with CD14+ magnetic beads, and CD14+ PBMCs were separated in a magnetic field; the sorted cells were added to DC culture bags with IL-4 (final concentration 30 ng / ml), GM-CSF (final Concentration is 60ng / ml) at 37°C, 5% CO 2 Co-cultivate in an incubator, the culture medium is PRMI1640 (containing 10% fetal bovine serum, 0.1ml penicillin-streptomycin), half of the culture medium is replaced every 3 days, and IL-4 and GM-CSF are added at the same time to induce dendrites generation of cells. After culturing for 10 days, cells that highly express HLA-I (77.31%), HLA-Dr (0.84%), CD80 (90.32%), CD86 (88.05%), and partially express CD40 (35.21%) were screened by flow cytometry , as the target DC cells.

[0068] Mi...

Embodiment 3

[0069] Example 3 Proliferative effect of DC cells loaded with pluripotent T cell epitope peptides on T cells

[0070] The WST-1 kit (ab155902, Abcam) was used to detect the stimulating effect of DC cells containing pluripotent T cell epitope peptides on the proliferation of T lymphocytes. The control group and the test group were respectively: DC cell group without antigenic peptide transformation, blank T cell group, DC cell group loaded with pluripotent T cell epitope peptide, and blank control group.

[0071] The specific measurement method is as follows:

[0072] Take a 96-well cell culture plate, add 0.1ml containing 2×10 5 RPMI1640 (containing 0.3% w / w PHA) culture medium of human peripheral blood T lymphocytes, at 37°C in 5% CO 2 After culturing for 2 hours, add 0.1ml (containing 2×10 4 cells) of DC cells loaded with pluripotent T cell epitope peptides, add 0.1ml (containing 2×10 4 cells) DC cells with untransformed antigen peptides, add 0.1ml (containing 2×10 cel...

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Abstract

The invention provides an SARS CoV-2 single-epitope T cell antigen peptide, a pluripotent T cell epitope peptide prepared from the antigen peptide, and a DC cell loaded with the pluripotent T cell epitope peptide; T cell immunity of an MHC-I pathway in vivo can be induced, and the immune function of an organism is reconstructed; and the antiviral function of autologous CD8 <+> CTL is stimulated, amore effective and safer means is provided for prevention and / or treatment of coronavirus infection related diseases, and wide clinical application and popularization prospects are achieved.

Description

technical field [0001] The present invention relates to the technical field of medicine, in particular to a pharmaceutical composition for preventing and / or treating coronavirus infection, a preparation method thereof, and an application thereof. Background technique [0002] Infection with SARS CoV-2 causes coronavirus disease 2019 (COVID-19), which can lead to death in severe cases. Effective therapeutic drugs against SARS CoV-2 infection have not yet been developed. If an effective SARSCoV-2 drug can be developed, it will surely play a key role in blocking the spread of the virus and controlling the outbreak. [0003] Up to now, several SARS CoV-2 vaccine projects have entered the clinical trial stage. The mRNA vaccine mRNA-1273 jointly developed by the National Institute of Allergy and Infectious Diseases under the National Institutes of Health and Moderna was approved by the FDA to enter clinical trials on March 5, 2020, and phase I began on March 16. Clinical Trials...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165C07K19/00C12N5/0784A61K39/215A61K35/17A61P31/14A61P11/00
CPCC07K14/005C12N5/0639A61K39/12A61K35/17A61P31/14A61P11/00C07K2319/00C12N2501/22C12N2501/2304C12N2501/998C12N2770/20022C12N2770/20034
Inventor 闫小君罗进
Owner 海图生物科技(上海)有限责任公司
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