Application of DHA (docosahexaenoic acid) or derivative thereof in preparation of coronavirus inhibitor
A technology of coronavirus and derivatives, applied in antiviral agents, antiviral immunoglobulins, medical preparations containing active ingredients, etc.
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Embodiment 1
[0053] Example 1, recombinant RBD protein preparation
[0054] 1.1. Construction of recombinant RBD protein expression vector
[0055] The coding sequence of the nucleotide design (the nucleotide sequence is SEQ ID No.2) of the receptor binding domain (RBD) protein is constructed in the pFastBac-Dual expression vector plasmid by homologous recombination, and the positive recombinant expression vector obtained is named pFastBac-RBD. The structure of the recombinant expression vector pFastBac-RBD is: the sequence between the polyclonal locus of the skeleton carrier pFastBac-Dual is replaced with a DNA molecule whose nucleotide sequence is SEQ ID No.2, and the sequence between the polyclonal locus of pFastBac-Dual is replaced, and the recombinant expression vector obtained is kept unchanged by the other nucleotide sequences of pFastBac.
[0056] The amino acid sequence of RBD is such as SEQ ID No.1, where SEQ ID No.1 is the signal peptide at positions 1-20, positions 21-214 are amino ...
Embodiment 2
[0062] Example 2, high-throughput screening found that DHA combined with RBD and the verification of the interaction between the two
[0063] 2.1. Surface plasmon resonance imaging experiments found that DHA binds RBD
[0064] We discovered that DHA can bind to RBD by means of SPRi surface plasmon resonance imaging technology. The specific experimental methods are as follows:
[0065] High-throughput detection of protein-small molecule affinity using the Plexera HT SPRi Biomolecular Interaction Analyzer. First of all, the arrayjet flying inkjet biochip dispensing platform is used to print the small molecule DMSO solution in the compound library in the form of a microarray on the surface of the small molecule optical crosslinking chip provided by Plexera, and after the chip is dried in vacuum, the small molecule is covalently fixed on the chip surface by ultraviolet excitation crosslinking reaction, and the excess small molecules are cleaned on the chip for detection. The RBD prote...
Embodiment 3
[0073] Example 3, DHA synergistic enhancement antibody inhibits the invasion of the new crown pseudovirus
[0074] The present invention is based on the new crown pseudoviral invasion system constructed by Zhang Linqi of Tsinghua University School of Medicine to detect the antiviral activity of DHA, and the virus preparation, antibody drug activity detection method and data processing method related to the system have been disclosed in the literature.
[0075] The results show that DHA and its derivatives, EDPs, can effectively enhance the effect of multiple new coronavirus neutralizing antibodies to neutralize the virus.
[0076] Taking the neutralizing antibodies numbered P2C-1F11 and P2B-1G5 as an example, the dose-efficiency curve shifts significantly to the left with the addition of DHA or DHA derivatives EDPs. In the case of IC50, the IC50 of P2C-1F11 was reduced from 0.039 μg / mL to 0.023 μg / mL (DHA) and 0.013 μg / mL (EDPs), respectively, and the IC50 of P2B-1G5 was reduced fr...
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