Oligopeptide and its derivative and application thereof
A derivative and oligopeptide technology, applied to oligopeptide and its derivatives and application fields, can solve the problems of difficult synthesis and large molecular weight, and achieve the effects of small molecular weight, inhibiting proliferation and inhibiting the activity of influenza virus
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Embodiment 1
[0035] Virus virulence assay (TCID 50 )
[0036] Influenza H1N1 (FM1), H3N2 (N3), H5N1 and H7N9 virus antigens used in the present invention are virus inactivated antigens with hemagglutination activity, all can cause disease, H1N1, H3N2 are human influenza, H5N1 and H7N9 It is bird flu. Taking H7N9 as an example, the virus virulence was tested to prove its activity. The specific experimental steps and results are as follows:
[0037] After the MDCK cells (influenza virus susceptible canine kidney passage cell strain) in the logarithmic growth phase are laid on a single layer, the influenza virus H1N1 is continuously diluted by 10 times, and 10 -2 ~10 -10 The diluted virus was inoculated into MDCK cells, adsorbed at 37°C for 1 hour, washed twice with PBS, and then replaced with maintenance medium to continue culturing. Normal cells were used as controls, and each well had 4 replicate wells. Observe the cell lesion under an inverted microscope every day, record the lesion de...
Embodiment 2
[0045]The present invention uses the protein database (Protein Data Bank) hemagglutinin protein (1HGG) crystal structure as the research basis, screens out a series of oligopeptides with antiviral activity, its sequence contains W-Arg-Pro-X-Y-Z domain, A class of oligopeptide derivatives is obtained by modifying the short peptide W-Arg-Pro-X-Y-Z with stearic acid. The general structural formula of this kind of oligopeptide derivatives is C 17 h 35 CO-W-Arg-Pro-X-Y-Z, wherein, W is a deletion or a hydrophobic amino acid (such as W is a deletion or Ala); X and Z are hydrophobic amino acids (such as X is Leu, Phe; Z is Ala, Leu ); Y is a positively charged amino acid (for example, Y is Lys, Arg). These oligopeptide derivatives all have antiviral activity and are expected to be developed into ideal antiviral drugs. The specific structure of oligopeptide derivatives can be:
[0046] Oligopeptide I, C 17 h 35 CO-Arg-Pro-Leu-Lys-Ala; or
[0047] Oligopeptide II, C 17 h 35 CO...
Embodiment 3
[0056] Anti-influenza virus efficacy test method:
[0057] Take cells in good logarithmic growth phase, add 0.25% trypsin digestion solution, blow evenly after digestion and fall off, count, and press 2×10 5 cells / well, seeded in 96-well cell culture plate, placed in constant temperature CO 2 Cultivate in the incubator for 16-24h. Set up normal cell control group, virus infection group, positive drug control group and test drug group respectively. Discard the culture medium, wash three times with PBS, add 100TCID to each well except the normal control group 50 Infectious amount of influenza virus, placed in constant temperature CO 2 The incubator was left to rest for 1 hour, unadsorbed virus was discarded, washed twice with PBS, and then drug-containing maintenance solutions with different concentrations were added. There were 6 doses in the test drug group, each with 3 replicate wells. Then continue to cultivate the cell culture plate with the above-mentioned scheme at 3...
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