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Method for preparing gene therapy medicine for novel corona virus 2019

A gene therapy and product technology, applied in the field of preparation of new coronary pneumonia gene therapy products, to achieve the effects of easy implementation, improved immunity, and novel technical solutions

Pending Publication Date: 2020-05-19
翁炳焕
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide an easy-to-implement and quick-acting method for preparing gene therapy products for COVID-19, so as to solve the problem that RNA interference technology cannot be applied to the treatment of COVID-19 in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Screening nCoV2019 siRNA sequences with high silencing efficiency against the whole nCoV2019 genome, using 4'-C-methyl-modified mononucleotide (AUCG) to synthesize nCoV2019 siRNA, and using ring-opening reaction to synthesize lipid-like epoxyalkylamine derivative EAAD , the lipid complex nanoparticle carrier LHNPs loaded with nCoV2019 siRNA by EAAD was prepared by nanoprecipitation method, the inner layer is polylactic acid glycolic acid PLGA containing EAAD and nCoV2019 siRNA conjugate, and the outer layer is wrapped with lipids that can improve the biocompatibility of the carrier The outermost layer is PEG that can improve the stability of the carrier structure. Finally, the carrier LHNPs and spray are formulated into a dosage form for the treatment of new coronary pneumonia through pulmonary inhalation, which is used to silence the homologous nCoV2019 gene, thereby producing anti-nCoV2019. effect.

[0024] 1. Preparation of phosphoramidite base monomer (AUCG)

[002...

Embodiment 2

[0079] 1. According to steps "1", "2" and "3" in Example 1, entrust a biological company to industrially synthesize nCoV2019 siRNA.

[0080] 2. Refer to "(4)" in step "4" in Example 1 to perform nCoV2019 siRNA test.

[0081] 3. Prepare nCoV2019siRNA dry powder inhalation according to step "5" in Example 1 for the treatment of novel coronavirus pneumonia. This Example 2 can prepare gene therapy products for novel coronavirus pneumonia more quickly than Example 1 .

Embodiment 3

[0083] 1. Design primers according to step "2" in Example 1, screen for one or several nCoV2019 targeting sequences with significant gene silencing effects, and entrust a biological company to industrially synthesize nCoV2019 siRNA.

[0084] 2. Refer to "(4)" in step "4" in Example 1 to perform nCoV2019 siRNA test.

[0085]3. The nCoV2019 siRNA industrially synthesized by the biological company is made into the following sprays for the treatment of new coronary pneumonia.

[0086] Prepare the nCoV2019 siRNA dry powder inhaler according to the "5" step in Example 1.

[0087] Mix synthetic nCoV2019 siRNA with water by siRNA:H 2 O=1:5-50 (mole or mass / volume) formulated as a spray for respiratory or oral administration.

[0088] Synthetic nCoV2019 siRNA and suitable existing medical sprays are formulated into sprays for respiratory or oral administration according to siRNA: existing medical sprays = 1:5-50 (molar or mass / volume).

[0089] Because the synthesis of genes o...

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Abstract

The invention relates to a method for preparing a gene therapy medicine for novel corona virus 2019. The method comprises the following steps: according to an nCoV2019 (novel corona virus 2019) genome, screening an siRNA (small interference ribonucleic acid) sequence with an RNA interference effect, and performing synthesis by using a nucleic acid synthesis method; and preparing synthesized nCoV2019siRNA into lipid nanoparticles LHNPs, and compounding LHNPs or nCoV2019siRNA with a spray, so as to obtain a gene therapy product with inhalation administration. When reaching target cells, siRNA isdegraded since an siRNA antisense strand which is generated under the action of inherent and stress boosted helicase and endonuclease / exonuclease is combined with homologous RNA and / or mRNA (messenger ribonucleic acid). In addition, type-I interferon, IL-6, TNFa (tumor necrosis factor a) and the like can be generated under stimulation of double-strand siRNA and cellular immunity can be mediated,it is known that type-I interferon has antiviral immunity, IL-6 is capable of inducing hyperplasia of immune globulin and T cells, and the TNFa is capable of promoting infection resistance of neutrophile granulocyte, inducing apoptosis of virus infected cells and thus inhibiting virus hyperplasia, so that in addition to virus prevention of RNA interference, antibacterial antiviral immunity of cellfactors, immune cells and immune globulin can be also triggered, and in addition, the medicine can be efficiently and rapidly synthesized, and is easy in short-term application to treatment or prevention.

Description

technical field [0001] The invention relates to a preparation method of a novel coronavirus pneumonia gene therapy product, belonging to the field of biological product preparation. Background technique [0002] Human coronaviruses (HcoV 229E and HcoV OC43) cause up to 30% of colds. Animal coronaviruses, such as porcine gastroenteritis coronavirus (TGEV), mouse hepatitis coronavirus (MHV), avian infectious bronchitis coronavirus (IBV), etc., can infect the respiratory tract, gastrointestinal tract, nervous system and liver of the corresponding host , resulting in corresponding symptoms. The envelope of the coronavirus has petal-shaped protrusions that make the coronavirus look like a crown (Latin, corona), and its nucleocapsid is a variable long helix. The virion diameter of the coronavirus is 60nm-140nm, spherical, and the virus genome is a single-stranded positive-sense RNA of 27kb-32kb, which is the largest among all RNA virus genomes. A variant of coronavirus caused t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/88A61K31/7105A61P31/14
CPCA61K31/7105A61P31/14C12N15/88
Inventor 翁炳焕李兰娟
Owner 翁炳焕
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