CN-121974939-A - Borated lipid and high-efficiency delivery system and preparation method thereof

Abstract

The invention relates to the technical field of medicines, in particular to a borated lipid and high-efficiency delivery system and a preparation method thereof. The borated lipid has a structure shown in a formula I. The high-efficiency delivery vector can realize stable encapsulation and accurate release of nucleic acid such as RNA molecules, and the like, shows remarkable targeted transmission advantage in a cell delivery experiment, and can lead RNA drugs to be specifically delivered to macrophages and other cells through Gao Xiaojie. Safety is improved and potential risks associated with conventional delivery systems are avoided.

Inventors

• DENG HONGZHANG
• ZHAO CAIYAN
• WANG RUJIE

Assignees

• 西安电子科技大学

Dates

Publication Date

20260505

Application Date

20251230

Claims (10)

1. 1. A borated lipid, characterized in that the borated lipid has the structure of formula I: I is a kind of Wherein R 1 and R 2 are each independently selected from a hydrocarbyl or hydrocarbylene-like structure providing the hydrophobic driving force for amphiphilic molecules, M1-M5 are each independently selected from one or more of an ester bond, an amide bond, an ether bond, an amine bond, a urea bond, a urethane bond, a carbon-carbon bond, a disulfide bond, or a hydrazone bond, X is a linking structure comprising A 1 , A 1 is hydrogen or is selected from a substituted or unsubstituted C 1 -C 20 alkyl, C 2 -C 20 alkenyl, C 2 -C 20 alkynyl, C 6 -C 30 aryl, or C 3 -C 30 heteroaryl, and L1 and L2 are each independently selected from an aryl or heteroaryl group comprising a boric acid group.
2. 2. The borated lipid of claim 1 wherein R 1 and R 2 are each independently selected from one or more of lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl, linoleoyl, n-dodecyl, n-hexadecyl, phytyl, farnesyl, cholesteryl, ergosterol or lanosterol groups, R 1 and R 2 are optionally substituted with fluoro, chloro, hydroxy or C1-C6 alkoxy, and/or wherein L 1 and L 2 each independently comprise a structure selected from phenylboronic acid, pyridinylboronic acid, pyrimidinylboronic acid or thiophenylboronic acid.
3. 3. A high efficiency delivery system comprising a borated lipid, a steroid and a PEG lipid, and optionally a nucleic acid, wherein the borated lipid is a phenylboronic acid and derivatives thereof functionalized lipid, preferably a phenylboronic acid functionalized lipid, more preferably a borated lipid of the structure of formula I as claimed in claim 1 or 2.
4. 4. The efficient delivery system of claim 3, wherein the mass ratio of borated lipid, steroid, and PEG lipid is 20-80:10-40:10-40.
5. 5. The high-efficiency delivery system of claim 3 or 4, wherein the steroid is selected from one or more of cholesterol, cholesterol esters, steroid hormones, steroid vitamins, and phytosterols, and/or the PEG-modified lipid is selected from polyethylene glycol-derived lipids.
6. 6. The efficient delivery system of any one of claims 3-5, wherein the nucleic acid comprises one or more of mRNA, siRNA, DNA and micrornas.
7. 7. The efficient delivery system of any one of claims 3-6, wherein the mass ratio of borated lipid to nucleic acid is 1-500:1.
8. 8. The method of preparing a high efficiency delivery system according to any one of claims 3 to 7, wherein the self-assembly of the boronated lipids, steroids and PEG lipids is performed, preferably the phenylboronic acid lipids, steroids, PEG lipids and nucleic acids are prepared by microfluidic techniques or dialysis or thin film or titration.
9. 9. The method of preparing a high efficiency delivery system of claim 8, comprising the steps of: 1) Mixing borated lipid compound, steroid and PEG lipid, and dissolving with organic solvent to obtain organic phase liposome solution; 2) Dissolving nucleic acid in buffer solution with pH value of 3.0-8.0 to obtain water phase nucleic acid solution; 3) Mixing the organic phase liposome solution with the aqueous phase nucleic acid solution according to the volume ratio of 1:1-10 is preferably carried out by adopting microfluidic treatment.
10. 10. Use of the borated lipid of claim 1 or 2 or the high efficiency delivery system of any one of claims 3 to 7, characterized by the use in at least one of the following: 1) For encapsulating a nucleic acid encoding a peptide or protein; 2) For use in a vaccine; 3) For use in the treatment or prophylaxis of a peptide or protein encoded by a nucleic acid as a medicament for the treatment or prophylaxis of a disease; 4) Application to nucleic acid drug delivery vehicles; 5) Can be used in cosmetics.

Description

Borated lipid and high-efficiency delivery system and preparation method thereof Technical Field The invention belongs to the technical field of medicines, and particularly relates to a borated lipid and high-efficiency delivery system and a preparation method thereof. Background In the field of contemporary biological medicine, nucleic acids such as RNA are leading to rapid changes in therapeutic approaches as a novel drug target with great potential. Among them, mRNA therapy has been the focus of global scientific and medical attention by virtue of its great potential in the treatment of numerous diseases. The marked outcome is that two new crown vaccines based on mRNA were successfully approved by the U.S. Food and Drug Administration (FDA), a breakthrough in progress as a catalyst, motivated intensive exploration and study of mRNA therapies. mRNA, which is a single-stranded RNA, has unique biological properties and can rapidly act upon entry into cells to produce a transient but effective therapeutic effect. Currently, mRNA-based cancer treatment protocols, therapies for genetic diseases, protein replacement therapies, and various vaccines have steadily entered clinical trial stages, and have shown broad application prospects. However, mRNA suffers from a number of troublesome problems in practical use. Firstly, the single-chain structure is poor in stability and is easily degraded by being attacked by nuclease in vivo, secondly, the mRNA has a large molecular weight, is usually in the range of 300kDa to 5000kDa and carries negative charges, and the physical and chemical characteristics seriously prevent the mRNA from entering target cells, thirdly, the mRNA serving as an exogenous molecule is easily identified by an immune system of an organism so as to induce immune response, and fourthly, even if the mRNA successfully enters the cells, the mRNA can be degraded by lysosomes if the mRNA can not reach cytoplasm in time, so that the therapeutic effect is greatly reduced. The development of suitable delivery vehicles is critical to achieving efficient delivery of mRNA. Conventional gene delivery vectors are largely divided into viral vectors and non-viral vectors. Viral vectors have achieved some success in clinical practice, but face a number of limitations. For example, an innate immune response of the host is triggered, there is a risk of virus-induced immunogenicity, potential genomic integration hazards, relatively limited gene loading, difficulty in achieving repeated dosing, susceptibility to multiple complications upon dose escalation, and high production costs. In view of this, non-viral vectors are becoming the focus of research, including polymers, liposomes, and Lipid Nanoparticles (LNPs), among others. These non-viral vectors have driven the development of mRNA delivery systems based on their construction. However, despite some breakthrough in gene delivery efficiency, there are a number of drawbacks to non-viral vectors. Common cationic polymer delivery vehicles, because of carrying a relatively high positive charge, may trigger hemolysis and inflammatory reactions, while most polymer backbones are composed of non-degradable carbon chains, which are prone to toxicity. Currently, the mRNA vaccines obtained in batches mainly use LNPs as delivery vehicle, but traditional LNPs is highly dependent on cationic lipid components, which are extremely prone to cause severe inflammatory reactions in vivo. It follows that the optimal design of gene delivery vectors still faces significant challenges. CN109152830a discloses a tumor vaccine based on core-shell structured nanoparticles, the main function of which is to deliver mRNA encoding tumor-specific antigens into antigen presenting cells. The preparation process includes combining positively charged polymer and mRNA encoding tumor antigen to form one stable core. Then, the cationic lipid, neutral lipid and PEG lipid are wrapped outside the core by using a microfluidic technology, and finally the tumor vaccine is prepared. CN114599375A discloses a gene delivery platform consisting of a folic acid-cationic oligopeptide complex, the cationic oligopeptide containing 8-40 amino acids with specific amino acid residue continuous portions. The siRNA and shRNA stability can be improved, and the targeting delivery to folic acid receptor cells can be realized. Currently, there is an urgent need in the art to develop a safe and effective novel nucleic acid delivery system that is less immunogenic, does not cause an inflammatory response, and protects the nucleic acid drug from damage. Disclosure of Invention In order to solve the technical problems, the invention provides a borated lipid and high-efficiency delivery system and a preparation method thereof. The invention discloses a non-ionic bond-based nucleic acid encapsulation liposome delivery carrier and a non-ionic bond-based nucleic acid encapsulation liposome delivery system, and aims to solve the key pro