Method for inhibiting coronavirus by anhydride-modified protein
A coronavirus and protein technology, applied in the field of biomedicine, can solve problems such as no new coronavirus specific drugs
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[0036] Methods for preparing formulations known in the art can be found, for example, in "Remington's Pharmaceutical Sciences" (19th Ed.), Ed. A. Gennaro, 1995, Mack Publishing Company, Easton, Pa. Formulations for parenteral administration may, for example, contain excipients, sterile water or saline, poly(alkylene) glycols such as polyethylene glycol, vegetable oils or hydrogenated naphthalenes. Biocompatible, biodegradable lactide polymers, lactide / glycolide copolymers, or polyoxyethylene-polyoxypropylene copolymers can be used to control the release of the compounds. Other potentially useful parenteral delivery systems include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Formulations for inhalation may contain excipients such as lactose, or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate and deoxycholate, or may be in the form of nasal drops or Oily solution applied a...
Embodiment 1
[0047] Embodiment 1: the preparation of the protein of acid anhydride modification
[0048] Preparation of 3HP-HSA
[0049] (1) 3-hydroxyl-phthalic anhydride (3HP) is formulated with dimethyl sulfoxide (DMSO) into a mother liquor with a concentration of 1M;
[0050] (2) Prepare a disodium hydrogen phosphate solution (pH8.5) with a concentration of 0.1M, add human serum albumin to make an HSA solution, and the HSA concentration is 20 mg / ml;
[0051] (3) Add the acid anhydride solution prepared in step (1) to the HSA solution to be modified, so that the final concentration of the added acid anhydride is 12mM, mix well, adjust the pH of the protein solution to 9.0 with 4M NaOH solution, and incubate at 25°C for 20min ;
[0052] (4) Repeat step (3) for a total of five times, each time the final concentration of acid anhydride is 12mM, 24mM, 36mM, 48mM, 60mM, that is, the final concentration of acid anhydride in the solution is 60mM, and incubate at 25°C for 2h to complete the ...
Embodiment 2
[0075] Embodiment 2: the effect of the protein modified by acid anhydride on coronavirus live virus
[0076] Preparation of pseudovirus
[0077](1) Codon optimization was performed on the envelope protein (S protein) gene sequence of the new coronavirus SARS-CoV-2, and gene synthesis was carried out. The information is as follows.
[0078] SARS-CoV-2S protein gene sequence (synthesized by GenScript Biotechnology Co., Ltd.):
[0079] ATGCCCATGGGGTCTCTGCAACCGCTGGCCACCTTGTACCTGCTGGGGATGCTGGTCGCTT CCGTGCTAGCCCAGTGCGTGAACCTGACCACAAGGACCCAGCTGCCCCCTGCCTATACCAATTC CTTCACACGGGGCGTGTACTATCCCGACAAGGTGTTTAGAAGCTCCGTGCTGCACTCTACACAG GATCTGTTTCTGCCTTTCTTTAGCAACGTGACCTGGTTCCACGCCATCCACGTGAGCGGCACCA ATGGCACAAAGCGGTTCGACAATCCAGTGCTGCCCTTTAACGATGGCGTGTACTTCGCCTCTAC CGAGAAGAGCAACATCATCAGAGGCTGGATCTTTGGCACCACACTGGACTCCAAGACACAGTC TCTGCTGATCGTGAACAATGCCACCAACGTCGTGATCAAGGTGTGCGAGTTCCAGTTTTGTAAT GATCCTTTCCTGGGCGTGTACTATCACAAGAACAATAAGAGCTGGATGGAGTCCGAGTTTCGCG TGTATTCTAGCGCCAACAATTGCACATTTGAGTACG...
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