CN-121987809-A - Method for preparing nucleic acid medicine based on biological ions and application of nucleic acid medicine

Abstract

The invention belongs to the field of biological medicine, relates to delivery and stability of nucleic acid medicines, and in particular relates to a method for preparing a nucleic acid medicine based on biological ions and application of the nucleic acid medicine. The invention provides a zwitterionic biological material which is a cationic polyelectrolyte-nucleic acid micelle formed by inorganic salt and nucleic acid, has biocompatibility, degradability and mild preparation conditions, can deliver the nucleic acid into organisms, and simultaneously is used as one of important plant nutrition sources, and has long coexistence history and supplement in the fields of human beings and ecological environment. The structural characteristics of the plant genetic transformation agent are further discussed, and the plant genetic transformation agent provides a scheme for the application of RNA delivery such as human drug mRNA, siRNA and the like, RNA biopesticide to multiple scenes such as agricultural pest control and the like.

Inventors

• LI HAICHAO
• GUAN RUOBING
• WANG YINUO
• MIAO XUEXIA

Assignees

• 上海植生优谷生物技术有限公司

Dates

Publication Date

20260508

Application Date

20260130

Claims (10)

1. 1. The application of the biological ion in preparing nucleic acid medicaments is characterized in that the biological ion is selected from one or more of sodium nitrite, sodium chloride, sodium sulfate, sodium selenite, sodium phosphate, ammonium nitrate, aluminum trichloride, ammonium persulfate, ammonium dihydrogen phosphate, ammonium chloride, potassium chloride, manganese sulfate, copper sulfate, cobalt chloride, potassium iodide, sodium silicate and magnesium chloride.
2. 2. The method of claim 1, wherein the nucleic acid drug is a nucleic acid micelle formed by the biological ions and the nucleic acid.
3. 3. The method of claim 2, wherein the nucleic acid is selected from the group consisting of DNA, dsRNA, ssDNA and ssRNA.
4. 4. A preparation method of a nucleic acid drug is characterized by comprising the steps of mixing a nucleic acid aqueous solution with a biological ion solution to obtain a BIs@dsRNA complex solution, namely the nucleic acid drug.
5. 5. The method for producing a nucleic acid drug according to claim 4, wherein the nucleic acid is any one selected from DNA, dsRNA, ssDNA and ssRNA, and the water is pure water free of a nuclease.
6. 6. The method for preparing a nucleic acid drug according to claim 4, wherein the biological ion is selected from one or more of sodium nitrite, sodium chloride, sodium sulfate, sodium selenite, sodium phosphate, ammonium nitrate, aluminum trichloride, ammonium persulfate, ammonium dihydrogen phosphate, ammonium chloride, potassium chloride, manganese sulfate, copper sulfate, cobalt chloride, potassium iodide, sodium silicate, and magnesium chloride, and the molar ratio of the biological ion to the nucleic acid is in the range of 1:10 -6 -1:10 -3 .
7. 7. The method of nucleic acid according to claim 6, wherein the molar ratio of any one of potassium iodide, aluminum trichloride and sodium phosphate to nucleic acid is 1:10 -6 , the molar ratio of any one of sodium nitrite, sodium chloride, sodium sulfate, sodium selenite, ammonium nitrate, ammonium persulfate, ammonium dihydrogen phosphate, ammonium chloride, potassium chloride, manganese chloride and copper sulfate to nucleic acid is 1:10 -6 -1 : 10 -5 , the molar ratio of sodium silicate to nucleic acid is 1:10 -4 , and the molar ratio of manganese sulfate, magnesium chloride to nucleic acid is 1:10 -6 -1 : 10 -4 .
8. 8. A nucleic acid drug prepared by the method according to any one of claims 4 to 7, wherein the nucleic acid drug is a cationic polyelectrolyte formed by biological ions and nucleic acids, i.e., a nucleic acid micelle.
9. 9. The nucleic acid drug according to claim 8, wherein the mass volume percentage of the biological ions in the nucleic acid drug is 0.1% -48%, and the mass volume percentage of the nucleic acid is 0.1% -10%; When the biological ion is one or more of sodium selenite, sodium phosphate, magnesium chloride, potassium chloride, sodium chloride and sodium sulfate, the particle size of the nucleic acid medicine is 60-114.4nm; When the biological ion is one or more of aluminum chloride, ammonium hydrogen phosphate, ammonium chloride and sodium silicate, the particle size of the nucleic acid medicine is 542-1057nm.
10. 10. Use of the nucleic acid drag of claim 8 for delivering a nucleic acid drag.

Description

Method for preparing nucleic acid medicine based on biological ions and application of nucleic acid medicine Technical Field The invention belongs to the field of biological medicine, and relates to delivery and stability of nucleic acid medicines. Background Plant genetic transformation is an important component of agricultural engineering, small molecule synthesis, and bioenergy work, and traditionally used plant DNA delivery tools limit the plant species that can be transformed or exhibit low transformation efficiency and tissue damage due to the use of high external forces. There is a need for new delivery materials that deliver genes and proteins into intact cells without external forces or assistance. Meanwhile, animal genetic transformation requires the addition of more new delivery means in terms of delivery efficiency, cytotoxicity and the like. In addition, the rapid development of mRNA vaccines and RNAi drugs in recent years, once again demonstrated the potential of RNA for use in organisms, a central issue in determining this potential is how to efficiently deliver RNA to the focal site of an organism and how to stably expose RNA molecules to the environment. The generation of nucleic acid delivery materials with various materials not only enables the production and application of RNA to be possible, but also provides a feasible basis for the development of RNA medicaments. Of course, their use faces many challenges concerning stability and efficacy, for which researchers have developed a variety of delivery materials such as liposomes, virus-like particles, composite nanoparticles, and bio-clays, among others. However, these delivery materials have problems of high cost, difficulty in preparation, unfriendly environment, and the like. Inorganic ions coexist in the field of human beings and ecological environment, have long history and supplement each other. Such as nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), calcium (Ca) in macroelements, iron (Fe), manganese (Mn), zinc (Zn), copper (Cu), boron (B), molybdenum (Mo), chlorine (Cl), sodium (Na), nickel (Ni) in microelements. NO 3- is the main form of plant nitrogen for synthesizing protein/chlorophyll, H 2PO4- is the only form of plant phosphorus for forming ATP/nucleic acid/phospholipid, SO 42- is the main form of plant sulfur for synthesizing sulfur-containing amino acid/coenzyme, cl - participates in osmotic adjustment/light cooperation, cell turgor maintenance and the like. Research shows that part of inorganic ions play an important role in genetic information transmission links such as DNA damage repair, RNA synthesis and the like, and the inorganic ions are possibly not fully discovered in nucleic acid metabolism and stability regulation. Based on the above, the research is to explore the interaction relation between inorganic ions and nucleic acid, and aims to discover the potential of the inorganic ions as a novel, green and efficient nucleic acid delivery and stabilization system, so as to provide innovative material basis and technical path for animal and plant genetic transformation, RNA drug delivery and agricultural biotechnology. Disclosure of Invention In order to solve the technical problems, the invention provides a method for preparing a nucleic acid medicine based on biological ions and application of the nucleic acid medicine. The technical scheme of the invention is realized as follows: In one aspect, the invention discloses application of Biological Ions (BIs) in preparing nucleic acid medicaments, wherein the Biological Ions (BIs) are selected from one or more of sodium nitrite (NaNO 2), hydrogen chloride (NaCl), sodium sulfate (Na 2SO4), sodium selenite (Na 2SeO3), sodium phosphate (Na 3PO4), ammonium nitrate (NH 4NO3), aluminum trichloride (AlCl 3), ammonium persulfate ((NH 4)2S2O8), ammonium dihydrogen phosphate (NH 4H2PO4), ammonium chloride (NH 4 Cl), potassium chloride (KCl), manganese chloride (MnCl 2), manganese sulfate (MnSO 4), cobalt chloride (CoCl 2), copper sulfate (CuSO 4), potassium iodide (KI), sodium silicate (Na 2SiO3) and magnesium chloride (MgCl 2). The nucleic acid drug is a nucleic acid micelle formed by biological ions and nucleic acid. After the biological ion composition is compounded with different Nucleic Acids (NA) such as DNA, dsRNA, ssDNA, ssRNA, the biological ion composition has an enzymolysis protection effect on the nucleic acids, and is beneficial to the delivery of the nucleic acids. In a second aspect, a method for preparing a nucleic acid drug is provided, comprising the steps of mixing an aqueous nucleic acid solution with a bioion solution to obtain a bis@dsRNA complex solution, namely the nucleic acid drug. The nucleic acid is selected from DNA, dsRNA, ssDNA or ssRNA, and water is pure water without nuclease. The biological ion is selected from one or more of sodium nitrite (NaNO 2), hydrogen chloride (NaCl), sodium sulfate (Na 2SO4), sodium selenite (Na 2SeO3), sod