Novel coronavirus subprotein nano vaccine and preparation method and application thereof
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A coronavirus and nano-vaccine technology, applied in the field of biomedicine, can solve the problems of difficulty in loading the new coronavirus S protein, inability to effectively large capacity, low immune response, etc., to achieve the prevention of new coronavirus infection, a new and efficient way and The effect of selection and efficient loading of antigen
Pending Publication Date: 2021-10-08
WUHAN SHENGRUN BIOTECHNOLOGY CO LTD
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[0003] In recent years, nano-vaccine platforms dedicated to the co-delivery of antigens and adjuvants have been developed rapidly. Liposomes, albumin, and some amphiphilic block copolymers that have passed the FDA are nanocarriers used in delivery platforms in recent years. Lipids Body refers to amphiphilic phospholipid molecules as the basic material, adding cholesterol and other auxiliary materials, in water, because the hydrophobic segments of amphiphilic phospholipids are gathered together, and the hydrophilic ends are exposed to water, thus self-assembling to form the middle hydrophobic and hydrophilic ends. Bilayer structure vesicles; albumin is the most abundant protein in plasma, using albumin as a carrier for insoluble drugs to prepare drug delivery nanoparticles will make it have low biotoxicity, low immune response, biodegradability and increase half-life and other advantages, the binding force between albumin and hydrophobic small molecule drugs is mainly van der Waals force and hydrogen bond force, hydrophobic small molecule drugs are mainly loaded with the IIA region of bovine
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Embodiment 1
[0040] The preparation of embodiment 1 novel coronavirus subprotein nano-vaccine
[0041] 1. Synthesis of polylactic acid PLA
[0042] With n-hexanol as the initiator, stannous octoate (Sn(Oct) 2 ) as a catalyst to synthesize PLA by the method of ring-opening polymerization of L-lactide (L-LA) and D-lactide (D-LA), and its synthetic route is as follows:
[0043] The specific operation steps are:
[0044] Weigh 0.2500g of n-hexanol in the glove box and place it in a dry ampoule, add 2.1180g each of L-LA and D-LA, and add 0.31mL of pre-configured Sn(Oct) with a concentration of 0.02g / mL 2 solution, and finally add 21.0mL of dry toluene. Polymerization at 130°C, N 2 React in the environment for 2 days. The solution was settled with 200 mL of glacial ether, and the obtained product was dried at room temperature for 24 h. Then the product was dissolved in N,N-dimethylformamide (DMF), dialyzed in DMF through a dialysis bag (MWCO=1000D) for 48 hours to remove a small amount o...
Embodiment 2
[0071] The effect verification of embodiment 2 nano vaccine NPQ-RBD and NPQ-RBD-AP
[0072] 1. Characterization of Nano Vaccines
[0073] Nano-vaccine NPQ-RBD and NPQ-RBD-AP were respectively prepared by the method described in Example 1, and the particle size was detected to be between 100-200nm, and the nanoparticles in this range can effectively penetrate lymphatic vessels and be drained to the lymph nodes.
[0074]In order to verify the stability of the nano-vaccine NPQ-RBD and NPQ-RBD-AP carrying RBD fragments, the amino acid fragments on the nano-NPQ-RBD were labeled with FITC molecules; the nano-particles were dialyzed in saline at 4°C, and the fractions were collected every day Solution, using a microplate reader to detect the relative FITC fluorescence intensity in the nanometer, and set the nanoparticle NP-RBD control group without adjuvant, and the group injected with only RBD antigen, the detection results are as follows Figure 4 shown.
[0075] The results sho...
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Abstract
The invention discloses a novel coronavirus subprotein nano vaccine and a preparation method and application thereof. The nano vaccine comprises: an organic compound self-assembled by polylactic acid, a porphyrin or a porphyrin derivative and a Co2+ ion conjugate; a novel coronavirus antigen protein; a vaccine adjuvant and lipid. According to the invention, a core of the synthesized organic compound is coated with the adjuvant, and the surface of the synthesized organic compound is efficiently loaded with the novel coronavirus antigen protein, so that a nano vaccine system for co-delivery of the novel coronavirus antigen protein and the vaccine adjuvant is implemented, and not only the immunogenicity of a novel coronavirus recombinant subprotein can be played to the greatest extent, but also the distribution condition of the nano vaccine in a living body can be traced through fluorescent molecules. In addition, the nano vaccine provided by the invention is also connected with polypeptide capable of specifically targeting antigen presenting cells, so that the nano vaccine is promoted to be massively ingested by DC cells, and the antiviral reaction is promoted. Meanwhile, the preparation method of the nano vaccine is simple and efficient, and a brand new way is provided for effective prevention of novel coronavirus infection.
Description
technical field [0001] The invention belongs to the field of biomedicine, in particular to a novel coronavirus subprotein nano-vaccine and its preparation method and application. Background technique [0002] Novel coronavirus pneumonia (COVID-19) is a respiratory disease caused by a new type of coronavirus (SARS-CoV-2). Its clinical manifestations range from mild flu-like symptoms to severe life-threatening pneumonia. Infectiousness and fatality. Its main structure includes spike protein (S, spike), envelope protein (E, envelope), membrane protein / matrix protein (M, membrane / matrix) and nucleocapsid protein (N, uncleocapsid). When the host is infected with the new coronavirus, the S protein will be cleaved by protease into S1 subunit and S2 subunit, and the receptor binding domain (RBD) of the S1 subunit will bind to angiotensin converting enzyme 2 (Angiotensin-converting enzyme 2) on the surface of the host cell. Converting enzyme 2, ACE2) binding, causing fever and pulm...
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