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CN-117263818-B - Cationic lipid compound, and preparation method and application thereof

CN117263818BCN 117263818 BCN117263818 BCN 117263818BCN-117263818-B

Abstract

The application relates to the field of drug carriers, and particularly discloses a cationic lipid compound, a preparation method and application thereof, wherein the cationic lipid compound comprises a compound shown in a formula I, n=1, 2,3 or 4, R1 and R2 are respectively and independently selected from secondary long-chain alkyl esters shown in a formula II, and n1=2, 4 or 6, n2=5, 7, 9 or 11 and n3=5, 7, 9 or 11. The cationic lipid compound, phospholipid, cholesterol and polyethylene glycol lipid are combined to prepare LNP carrier, and then nucleic acid medicine is wrapped to prepare corresponding medicine. The cationic lipid compound has the advantages of easily available production raw materials, simple synthesis steps, good drug delivery effect, high-efficiency entry of corresponding drugs into receptor cells, activation of immune response of organisms and practical application value.

Inventors

  • DAI YONGCHAO
  • XU YUHONG

Assignees

  • 杭州高田生物医药有限公司

Dates

Publication Date
20260508
Application Date
20220614

Claims (12)

  1. 1. A cationic lipid compound, characterized in that the cationic lipid compound comprises a compound represented by formula I; in formula I, n=1, 2, 3 or 4; r 1 and R 2 are each independently selected from secondary long chain alkyl esters of formula II; In formula II, n 1 ＝6;n 2 ＝7;n 3 =7.
  2. 2. The cationic lipid compound of claim 1, wherein the cationic lipid compound comprises a compound of formula V;
  3. 3. the method for preparing a cationic lipid compound according to claim 1 or 2, wherein the method for preparing a cationic lipid compound comprises the steps of: the brominated ester compound is used as a raw material to sequentially carry out alkylation reaction and reduction reaction, Then the cationic lipid compound and acyl chloride compound undergo condensation reaction, two-step substitution reaction and one-step hydrolysis reaction in sequence, and finally undergo esterification reaction with a monobasic secondary alcohol compound to prepare the cationic lipid compound.
  4. 4. An LNP vector comprising any one or a combination of at least two of the cationic lipid compounds of claim 1.
  5. 5. The LNP carrier of claim 4 wherein the LNP carrier comprises 15% -70% cationic lipid compound in mole fraction.
  6. 6. The LNP carrier of claim 4 further comprising, in mole fraction, 8% -30% phospholipids, 15% -65% cholesterol and 1.5% -3% polyethylene glycol lipids.
  7. 7. A medicament comprising the LNP vector of any one of claims 4-6.
  8. 8. The medicament of claim 7, wherein the medicament further comprises a nucleic acid medicament.
  9. 9. The drug of claim 8, wherein the molar ratio of the nitrogen content of the cationic lipid compound in the LNP carrier to the phosphorus content of the nucleic acid drug is (2-6): 1.
  10. 10. A method of preparing a medicament according to any one of claims 7 to 9, comprising the steps of: weighing other components except the cationic lipid compound, dissolving, and preparing a stock solution; adding a cationic lipid compound into the stock solution, and mixing to obtain a lipid alcohol phase; Diluting nucleic acid medicine to prepare nucleic acid water phase; mixing the lipid alcohol phase with nucleic acid water to obtain intermediate product, and dialyzing to obtain the final product.
  11. 11. A vaccine comprising the LNP vector of any one of claims 4-6.
  12. 12. Use of a cationic lipid compound according to claim 1 or 2 in the preparation of an LNP vector, a medicament or a vaccine, or use of an LNP vector according to any one of claims 4-6 in the preparation of a medicament and/or a vaccine.

Description

Cationic lipid compound, and preparation method and application thereof Technical Field The application relates to the technical field of drug carriers, in particular to a cationic lipid compound, a preparation method and application thereof. Background Gene therapy (GENE THERAPY) is a new technique which is produced by combining modern medicine and molecular biology, and is characterized in that a target gene is introduced into a patient body to correct or compensate diseases caused by defective and abnormal genes, so that the diseases are treated. Gene therapy has been used successfully as a new means for disease treatment, and the scientific breakthrough achieved will continue to drive the development of gene therapy to mainstream medical treatment. Common genetic drugs are plasmid DNA (PLASMID DNA, PDNA), antisense oligonucleotides (ANTISENSE ODN), small interfering RNAs (siRNA), small hairpin RNAs (shRNA) and messenger RNAs (mRNA). mRNA is a single-stranded ribonucleic acid containing specific genetic information, which is capable of transferring the carried genetic information to ribosomes within cells where it serves as a template to direct the body to synthesize a specific protein of interest. The mechanism of action of mRNA enables delivery of transmembrane or intracellular proteins to the body compared to traditional protein drugs that have difficulty crossing cell membranes, thus enabling the treatment or prevention of more diseases. The above-described advantages of mRNA also make it a trend for future drug development. The key to gene therapy is to deliver the gene drug to the target cells in vivo to allow it to function. However, the foreign gene is directly introduced into the body, degraded by nuclease in the body, and degraded into small molecular nucleotides before entering the target cell, thereby losing therapeutic effect. Therefore, the key to achieving gene therapy is to construct an efficient, safe gene delivery system. Gene vectors undergo a number of complex processes in the delivery of genes, including reaching target cells through blood circulation, cellular uptake, endosomal escape, intracellular movement, and release of genetic material from the vector, with the major barriers being extracellular barriers to the complex blood environment and intracellular barriers to lysosomal enzymatic degradation. Therefore, how to find a good gene vector so that the target gene reaches the target spot to exert the effect is a problem to be solved urgently by a gene vector researcher. At present, gene delivery vector systems are mainly divided into two main categories, namely a viral vector system and a non-viral vector system. The virus vector is a natural vector resource, has simple structure, high transfection efficiency and strong target cell specificity, but has poor guidance and carrying capacity, immunogenicity and potential tumorigenicity, so that the virus genome is difficult to meet the requirements of clinical application. Therefore, non-viral vector systems that are diverse, non-immunogenic and easy to control production have been of great interest in recent years and have found application in many therapeutic fields. Commonly used non-viral vector systems are mainly lipid vectors. Lipid carriers typically contain the positive charge of a cationic lipid and bind to a negatively charged genetic drug by electrostatic interactions, thereby concentrating and packaging the genetic material into smaller particle size particles to form Lipid Nanoparticles (LNP). LNP shows incomparable advantages in the preparation of gene delivery vectors and cell transfection with other types of liposomes. The smaller particle size of the compound reduces the opportunities of recognition, phagocytosis and elimination by the in vivo macrophages, and improves the in vivo bioavailability of the medicine. Meanwhile, aiming at tumor tissues, the smaller particle size of the compound is easier to penetrate from vascular endothelial cell gaps to enter tumor parenchyma through permeation and retention effects, so that drug aggregation in the tumor tissues is increased. In the aspect of transfection, the cell surface is slightly negatively charged, so that the positively charged liposome is more easily adsorbed on the cell surface and enters cells through mechanisms such as endocytosis, and the transfection capacity of the liposome is greatly improved. At present, LNP becomes the most widely applied non-viral vector due to the characteristics of simple structure, simple and convenient operation, high biological safety and the like, but most LNP has complex preparation process and is not easy to carry out amplified production. Therefore, how to provide a non-viral vector system with simple preparation method and good therapeutic effect has become a problem to be solved. Disclosure of Invention In order to improve the drug delivery effect of the gene vector and simplify the synthesis steps thereof, the present appl