Ionizable lipid compound and composition thereof

Abstract

The invention discloses an ionizable lipid, a pharmaceutical composition containing the ionizable lipid and application of the ionizable lipid, and belongs to the technical field of biology. The invention provides a compound as shown in formula (I), or a pharmaceutically acceptable salt or a deuterated compound thereof, and the compound is used as an ionizable lipid, so that the technical defect that other ionizable lipid compounds in the prior art are mainly delivered to liver expression antigen protein and cannot be delivered to the spleen is overcome; the compound as shown in the formula (I) has the characteristics of good spleen targeting property, high delivery efficiency and the like; the compound shown in the formula (I) can be used for preparing lipidosome, lipid nanoparticles, drug carriers or compounds and is used for delivering nucleic acid drugs.

Description

Technical Field

[0001] This invention belongs to the field of biochemistry technology, specifically, it relates to an ionizable lipid compound and its composition. Background Technology

[0002] mRNA vaccines are a new type of nucleic acid vaccine that have demonstrated significant advantages in rapidly and effectively combating COVID-19 infection. The working principle of mRNA vaccines involves directly introducing mRNA encoding a specific antigen protein into somatic cells. The host cell's expression system then synthesizes the antigen protein, inducing the host's immune system to produce various effects, including B and T cell-specific immune responses to the antigen, effectively treating and preventing the disease. Compared to DNA nucleic acid vaccines and traditional protein / peptide vaccines, mRNA vaccines offer advantages such as rapid development, simple production processes, and high safety, making them a promising new type of nucleic acid vaccine for clinical application. The mRNA delivery vector is closely related to the vaccine's storage stability and immunogenicity, and is a core technology in mRNA vaccine development. Currently, existing nucleic acid delivery systems mainly fall into two categories: viral vectors and non-viral vectors. Compared to viral vectors, non-viral vectors, represented by liposomes, are considered superior nucleic acid delivery systems due to their low immunogenicity, good biocompatibility, and high transfection efficiency.

[0003] Currently, existing mRNA vaccines use LNPs as delivery vectors. LNPs utilize ionizable lipids to deliver mRNA to the liver for antigen protein expression. While this can activate the body's immune response, it still suffers from insufficient immunogenicity, leading to poor immunogenicity. Furthermore, for mRNA vaccines to exert stronger immunogenicity, the optimal target organ is an immune organ such as the spleen; however, other ionizable lipid compounds disclosed in existing technologies primarily deliver antigen proteins to the liver, also resulting in insufficient immunogenicity.

[0004] Therefore, there is a need to develop ionizable lipid compounds with high delivery efficiency, good immunogenicity, and good spleen targeting. Summary of the Invention

[0005] This invention aims to at least partially address one of the technical deficiencies existing in the prior art. To this end, this invention provides a novel ionizable lipid compound that, as an ionizable lipid, possesses characteristics such as good organ targeting and high delivery efficiency.

[0006] The first aspect of this invention provides a compound of formula (I), a pharmaceutically acceptable salt, or a deuterated compound:

[0007]

[0008] in,

[0009] X is any integer from 0 to 3;

[0010] Y is any integer from 1 to 3;

[0011] A is a substituted or unsubstituted C0-C3 alkylene group;

[0012] B is a substituted or unsubstituted C0-C5 alkylene group;

[0013] D is a substituted or unsubstituted C1-C5 alkylene group;

[0014] E represents substituted or unsubstituted C1-C. 20 Straight-chain or branched alkyl groups;

[0015] M is -O-, -S-, or -NH-;

[0016] G is -OC(=O)-, -C(=O)O-, -C(=O)N(H)- or -(H)NC(=O)-;

[0017] R1, R2, R3, and R4 are each independently selected from H, OH, -SH, =O, and NR5R6, wherein at least two of R1, R2, R3, and R4 are not simultaneously H;

[0018] R5 and R6 are each independently selected from H, OH, SH, and C1-C3 alkyl groups;

[0019] In a specific embodiment of the present invention, in the compound of formula (I), when X is 1, 2, or 3; when two of R1, R2, R3, and R4 are H, it means that two adjacent compounds are not simultaneously H;

[0020] In a specific embodiment of the present invention, in the compound of formula (I), R1, R2, R3, and R4 are selected from -OH and -SH;

[0021] In a specific embodiment of the present invention, in the compound of formula (I), X is 1;

[0022] In a specific embodiment of the present invention, in the compound of formula (I), Y is 1 or 2;

[0023] In a specific embodiment of the present invention, in the compound of formula (I), Y is 1;

[0024] In a specific embodiment of the present invention, M is -O- in the compound of formula (I);

[0025] In a specific embodiment of the present invention, in the compound of formula (I), M is -NH-;

[0026] In a specific embodiment of the present invention, in the compound of formula (I), M is -S-;

[0027] In a specific embodiment of the present invention, in the compound of formula (I), R1, R2, R3, and R4 are -OH;

[0028] In a specific embodiment of the present invention, in the compound of formula (I), G is -OC(=O)- or -C(=O)O-;

[0029] In a specific embodiment of the present invention, in the compound of formula (I), G is -C(=O)N(H)- or -(H)NC(=O)-; in a specific embodiment of the present invention, in the compound of formula (I),

[0030] X is 1;

[0031] Y is 1;

[0032] A is a non-substituted C0-C3 alkylene group;

[0033] B is a non-substituted C0-C5 alkylene group;

[0034] D is a non-substituted C1-C3 alkylene group;

[0035] M is O;

[0036] R1, R2, R3, and R4 are each independently selected from OH and -SH;

[0037] In a specific embodiment of the present invention, in the compound of formula (I),

[0038] X is 1;

[0039] Y is 2;

[0040] A is a non-substituted C0-C3 alkylene group;

[0041] B is a non-substituted C0-C5 alkylene group;

[0042] D is a non-substituted C1-C3 alkylene group;

[0043] M is O;

[0044] R1, R2, R3, and R4 are each independently selected from OH and -SH;

[0045] In a specific embodiment of the present invention, in the compound of formula (I),

[0046] X is 1;

[0047] Y is 1;

[0048] A is a non-substituted C0-C2 alkylene group;

[0049] B is a non-substituted C1-C2 alkylene group;

[0050] D is a non-substituted C1-C3 alkylene group;

[0051] M is O;

[0052] R1, R2, R3, and R4 are OH;

[0053] In a specific embodiment of the present invention, in the compound of formula (I),

[0054] X is 1;

[0055] Y is 1;

[0056] A is a non-substituted C0-C3 alkylene group;

[0057] B is a non-substituted C1 alkylene group;

[0058] D is a non-substituted C1-C3 alkylene group;

[0059] M is O;

[0060] R1, R2, R3, and R4 are OH;

[0061] In a specific embodiment of the present invention, in the compound of formula (I),

[0062] B is a C0 alkylene group;

[0063] R1, R2, R3, and R4 are equal to 0;

[0064] In a specific embodiment of the present invention, in the compound of formula (I),

[0065] E represents substituted or unsubstituted C5-C. 18 Straight-chain or branched alkyl groups;

[0066] In a specific embodiment of the present invention, in the compound of formula (I),

[0067] E represents non-substituted C5-C. 18 Branched alkyl groups;

[0068] Furthermore, in a specific embodiment of the present invention, in the compound of formula (I), E is selected from the following groups:

[0069]

[0070]

[0071] In a specific embodiment of the present invention, in the compound of formula (I), X is 1;

[0072] Y is 1 or 2

[0073] A is -CH2-;

[0074] B is -CH2-;

[0075] D is -CH2- or -CH2CH2-;

[0076] M is O;

[0077] R1, R2, R3, and R4 are OH;

[0078] G is -OC(=O)-, -C(=O)O-;

[0079] E is

[0080]

[0081] In a specific embodiment of the present invention, in the compound of formula (I), X is 1;

[0082] Y is 1 or 2

[0083] A is -CH2-;

[0084] B is -CH2-;

[0085] D is -CH2- or -CH2CH2-;

[0086] M is O;

[0087] R1, R2, R3, and R4 are OH;

[0088] G is -C(=O)N(H)- or -(H)NC(=O)-;

[0089] E is

[0090]

[0091] The compounds of the present invention, or their pharmaceutically acceptable salts or deuterated compounds, wherein when X is 0, the atoms on both sides of X are directly bonded together;

[0092] The compounds of the present invention, or their pharmaceutically acceptable salts or deuterated compounds, wherein when A and B are CO, the atoms on both sides of A and B are directly connected; for example, when A is CO, the N atom on the left side of A and the carbon atom on the right side of A are directly connected.

[0093] The compounds of the present invention, or their pharmaceutically acceptable salts or deuterated compounds, are selected from the following structures:

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102] A second aspect of the present invention provides a composition comprising the compound described in the first aspect of the present invention or a pharmaceutically acceptable salt or deuterated compound thereof; wherein the composition further comprises a pharmaceutically active molecule and a pharmaceutically acceptable excipient;

[0103] Furthermore, the composition comprises at least one excipient selected from neutral phospholipids, steroids, and polyethylene glycol lipids.

[0104] Furthermore, in the composition provided by the present invention, the active pharmaceutical molecule is selected from DNA, ASO, siRNA, miRNA, mRNA, ribozyme, nucleic acid aptamer or a combination thereof; preferably mRNA.

[0105] Furthermore, in the composition, the compound of the first aspect of the present invention accounts for 30% to 60% of the composition in molar proportion;

[0106] Furthermore, in the composition, the compound of the first aspect of the present invention accounts for 40% to 60% of the composition in molar proportion;

[0107] Furthermore, in the composition, the compound of the first aspect of the present invention accounts for 45% to 55% of the composition in molar proportion;

[0108] In a specific embodiment of the present invention, the composition comprises: (i) 30% to 60% molar of the ionizable lipid compound described in the first aspect of the present invention, (ii) 5-25% molar of neutral lipids, (iii) 25-55% molar of steroids or steroid analogs, and (iv) 0.5-10% molar of PEG-lipids.

[0109] Furthermore, the neutral lipids are selected from one or more of the following: DOPE, DOPC, EPC, PC, HSPC, and DPPC;

[0110] Furthermore, the steroid is selected from one or more of the following: cholesterol, lanosterol, sitosterol, stigmasterol, and ergosterol;

[0111] Furthermore, the composition is prepared into lipid nanoparticles.

[0112] In another aspect, the present invention provides the use of the aforementioned first aspect ionizable lipid compound and second aspect composition in the preparation of liposomes, lipid nanoparticles, drug carriers or complexes.

[0113] According to embodiments of the present invention, the above-mentioned compounds have ionizable properties and can be used to prepare liposomes and lipid nanoparticles. The prepared liposomes or lipid nanoparticles can be used as drug carriers to form nucleic acid drug-liposome complexes.

[0114] In some embodiments, the above-mentioned compounds can be used to prepare liposomes in polymer form; the above-mentioned compounds can be used to prepare liposomes by forming covalent links with other substances; the above-mentioned compounds can be used to prepare liposomes by chemically reacting with other substances.

[0115] According to embodiments of the present invention, the specific method of preparing liposomes using the above-mentioned compounds is not limited. The use of the above-mentioned compounds to prepare liposomes, and the presence of all or part of the structure of the above-mentioned compounds in the liposomes, is considered an application of the present invention.

[0116] For application purposes, the compounds of the first aspect of this invention (typically in the form of liposomes combined with a bioactive ingredient) can be applied as crude chemicals or formulated as pharmaceutical compositions. The pharmaceutical compositions of this invention comprise a compound of structure (I) and one or more pharmaceutically acceptable carriers, diluents, or excipients. The compound of structure (I) is present in the composition in an amount that effectively forms liposomes and delivers the bioactive ingredient, for example, for treating a specific disease or condition. Appropriate concentrations and dosages can be readily determined by those skilled in the art.

[0117] In another aspect, the present invention proposes liposomes or lipid nanoparticles comprising the aforementioned compounds of the first aspect. According to embodiments of the present invention, the liposomes or lipid nanoparticles of the present invention can be effectively delivered to organs such as the heart, liver, spleen, lungs, and kidneys, especially to the spleen, effectively activating the body's immunity and increasing the level of specific antibodies in the animal's body.

[0118] In another aspect, the present invention provides a drug carrier. According to embodiments of the present invention, the drug carrier comprises the aforementioned compound or the aforementioned liposomes or lipid nanoparticles. According to embodiments of the present invention, the aforementioned compound is an ionizable lipid, the drug carrier is an ionizable carrier, and the aforementioned compound, liposomes, or lipid nanoparticles can be used to load drugs and deliver drugs into cells.

[0119] In another aspect, the present invention provides a complex. According to embodiments of the present invention, the complex comprises the aforementioned compound, or comprises the aforementioned liposomes, lipid nanoparticles, or drug carriers; and a bioactive ingredient. As mentioned above, the aforementioned compound is an ionizable lipid, and the drug carrier is an ionizable carrier. Thus, drugs can be loaded using the aforementioned compound, liposomes, or lipid nanoparticles, and a complex containing a bioactive ingredient can be prepared, which can deliver the bioactive ingredient into the body's cells for the treatment of diseases.

[0120] In another aspect, the present invention provides a pharmaceutical composition. According to embodiments of the present invention, the pharmaceutical composition comprises the aforementioned compound, or comprises the aforementioned liposomes or lipid nanoparticles, or the aforementioned drug carrier, or the aforementioned complex. As described above, the aforementioned compounds, liposomes or lipid nanoparticles, drug carriers, and complexes have advantages such as strong delivery capability and good immune activation effect. Loading bioactive components onto the aforementioned compounds, liposomes or lipid nanoparticles, or drug carriers can deliver the bioactive components into the body, which is beneficial for the loaded bioactive components to exert their therapeutic effects and for the treatment of diseases.

[0121] In some embodiments, the pharmaceutical composition also includes pharmaceutically acceptable excipients.

[0122] In another aspect, the present invention provides a pharmaceutical use. According to embodiments of the present invention, the present invention provides the use of the aforementioned compounds, or liposomes or lipid nanoparticles, or drug carriers, or complexes, or pharmaceutical compositions comprising the aforementioned compounds in the preparation of a medicament. As described above, the aforementioned compounds, liposomes or lipid nanoparticles, drug carriers, and complexes have advantages such as low cytotoxicity, good biocompatibility, strong delivery capability, and good immune activation effect. Loading bioactive components onto the aforementioned compounds, liposomes, or drug carriers can deliver the bioactive components into the body, which is beneficial for the loaded bioactive components to exert their therapeutic effects and for the treatment of diseases.

[0123] In a final aspect, the present invention provides a method for preparing the compound of the first aspect. According to embodiments of the present invention, the present invention provides a method for preparing the aforementioned compound.

[0124] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0125] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0126] Figure 14N4R@mRNA and S4N3@mRNA particle size and potential diagram;

[0127] Figure 24N4R@mRNA particle size and polydispersity index changes after storage at different temperatures for 1, 8, 13 and 36 days;

[0128] Figure 34 shows actual images of N4R@mRNA after being stored at different temperatures for 7, 14 and 36 days and then expressed in vivo.

[0129] Figure 44 shows the luminescent images and statistical graphs of intracellular expression of N4R@mRNA at different doses;

[0130] Figure 5. In vivo expression distribution of 4N4R@mRNA and 4N-lipid@mRNA, and statistical graph; Detailed Implementation

[0131] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0132] It should be noted that the structural and chemical formula descriptions in the embodiments or implementations of this invention are intended to cover all alternatives, modifications, and equivalent technical solutions, all of which are included within the scope of this invention. Those skilled in the art should recognize that many similar or equivalent methods and materials can be used to practice this invention. This invention is by no means limited to the methods and materials described herein. In the event that one or more of the linked documents, patents, and similar materials differ from or contradict this application (including but not limited to defined terminology, terminology application, described techniques, etc.), this invention shall prevail.

[0133] It should be further appreciated that certain features of the invention, for clarity, have been described in multiple independent embodiments or implementations, but may also be provided in combination in a single embodiment or implementation. Conversely, various features of the invention, for brevity, have been described in a single embodiment or implementation, but may also be provided individually or in any suitable sub-combination.

[0134] Definitions and explanations:

[0135] Unless otherwise indicated, the technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains, and unless otherwise indicated, all patent publications cited in the entirety of this disclosure are incorporated herein by reference.

[0136] Unless otherwise stated, the following definitions will apply in this invention. For the purposes of this invention, chemical elements are defined according to the periodic table, CAS version, and the Chemical Handbook, 75th Ed, 1994. Furthermore, general principles of organic chemistry are found in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007; therefore, all contents of this invention are incorporated herein by reference.

[0137] In this document, the terms “comprising” or “including” are open-ended expressions, meaning that they include the contents specified in this invention, but do not exclude other aspects.

[0138] This document also includes isotopically labeled compounds of the present invention that are identical to those compounds described herein except that one or more atoms are replaced by atoms with atomic masses or mass numbers different from those commonly found in nature. Exemplary isotopes that may also be introduced into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as... 2 H, 3 H, 13 C 14 C 15 N、 16 O、 17 O、 31 P, 32 P, 36 S, 18 F and 37 Cl.

[0139] Compounds of the present invention comprising other isotopes of the aforementioned isotopes and / or other atoms, as well as pharmaceutically acceptable salts of said compounds, are included within the scope of this invention. Isotope-labeled compounds of the present invention, such as radioactive isotopes, are also included. 3 H and 14 The incorporation of tritium into the compounds of this invention can be used for drug and / or substrate tissue distribution analysis. Due to its ease of preparation and detection, tritium-substituted compounds... 3 H, and carbon-14, i.e. 14 C isotopes are particularly preferred. In addition, heavier isotopes, such as deuterium, are used. 2 H substitution can offer therapeutic advantages stemming from greater metabolic stability, such as increased in vivo half-life or reduced dose requirements. Therefore, it may be preferred in some cases.

[0140] The stereochemical definitions and conventions used in this invention are generally in accordance with S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereochemistry of Organic Compounds,” John Wiley & Sons, Inc., New York, 1994. The compounds of this invention may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of this invention, including but not limited to diastereomers, enantiomers, and atropisomers, and mixtures thereof such as racemic mixtures, are also included within the scope of this invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral centers (or multiple chiral centers) in the molecule. The prefixes d and l, or (+) and (-), are symbols used to specify the plane-polarized rotation caused by a compound, where (-) or l indicates that the compound is levorotatory. Compounds prefixed with (+) or d are dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be called enantiomers, and mixtures of such isomers are generally called mixtures of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may occur when there is no stereoselectivity or stereospecificity in a chemical reaction or method.

[0141] Depending on the choice of raw materials and methods, the compounds of this invention can exist as one or a mixture of possible isomers, for example, as pure optical isomers, or as mixtures of isomers, such as as racemic and non-corresponding isomer mixtures, depending on the number of asymmetric carbon atoms. Optically active (R)- or (S)- isomers can be prepared using chiral synthons or chiral formulations, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be E or Z configuration; if the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituent may be cis or trans (cis- or trans-) configuration.

[0142] The compounds of this invention may contain asymmetric or chiral centers, and thus exist in different stereoisomer forms. It is contemplated that all stereoisomer forms of the compounds of this invention, including but not limited to diastereomers, enantiomers, atropisomers, and geometric (or conformational) isomers and mixtures thereof, such as racemic mixtures, are within the scope of this invention.

[0143] Unless otherwise stated, the structures described in this invention also represent all isomers (e.g., enantiomers, diastereomeric atropisomers, and geometric (or conformational) forms) including this structure; for example, R and S configurations of each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, individual stereochemical isomers of the compounds of this invention, as well as mixtures of enantiomers, diastereomeric mixtures, and mixtures of geometric isomers (or conformational isomers), are all within the scope of this invention.

[0144] Any asymmetric atom (e.g., carbon, etc.) in the compounds of the present invention may exist in a racemic or enantiomerically enriched form, such as (R)-, (S)-, or (R,S)- configuration. In some embodiments, each asymmetric atom has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% enantiomeric excess in the (R)- or (S)- configuration. If possible, substituents on atoms having unsaturated double bonds may be present in cis-(Z)- or trans-(E)- form.

[0145] Therefore, as described in this invention, unless otherwise specified, the compounds of this invention may exist in one or a mixture of possible isomers (including cis and trans isomers, optical isomers (e.g., R and S corresponding isomers), diastereomers, geometric isomers, rotational isomers, and shunting isomers), for example, in essentially pure geometric (cis or trans) isomers, diastereomers, optical isomers (enantiomers), racemates, or mixtures thereof.

[0146] In this article, solid lines can be used. solid wedge Or virtual wedge Describe the carbon-carbon bonds of the compounds of this application. Solid lines are used to depict bonds to asymmetric carbon atoms, indicating all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) at that carbon atom. Solid or imaginary wedges are used to depict stereoisomers indicated by the presence of bonds to asymmetric carbon atoms. When present in racemic mixtures, solid and imaginary wedges are used to define relative stereochemistry, not absolute stereochemistry.

[0147] Any mixture of isomers can be separated into pure or substantially pure geometric or optical isomers, diastereomers, and racemates based on the physicochemical differences of the components, for example by chromatography and / or stepwise crystallization.

[0148] Racemic mixtures of any resulting end product or intermediate can be separated into optical enantiomers using known methods, such as by separating their diastereomeric salts. Racemic products can also be separated by chiral chromatography, such as high-performance liquid chromatography (HPLC) using chiral adsorbents. In particular, enantiomers can be prepared by asymmetric synthesis (e.g., Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis (2nd Ed. Robert E. Gawley, Jeffrey Aubé, Elsevier, Oxford, UK, 2012); Eliel, ELStereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, SH Tables of Resolving Agents and Optical Resolutions p. 268 (ELEliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972).

[0149] In this paper, the term "tautomer" or "tautomer form" refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerization is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (also known as prototropic tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions via the rearrangement of some bonding electrons. A specific example of a keto-enol tautomer is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one. Another example of tautomerism is phenol-keto tautomerism. A specific example of a phenol-keto tautomer is the interconversion between pyridine-4-ol and pyridine-4(1H)-keto. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0150] In this document, the term "solvent" refers to an association formed by one or more solvent molecules with the compounds of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed by solvent molecules that are water.

[0151] In this document, the term "pharmaceutically acceptable" means that a substance or composition must be chemically and / or toxicologically compatible with other components of the formulation and / or the mammals to which it is treated.

[0152] In this document, the term "pharmaceutically acceptable salt" refers to the organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well-known in the field, as described in the literature: SMBerge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19. Salts formed from pharmaceutically acceptable non-toxic acids include, but are not limited to, inorganic acid salts (such as hydrochlorides, hydrobromic acids, phosphates, sulfates, and perchlorates) and organic acid salts (such as acetates, oxalates, maleates, tartrates, citrates, succinates, and malonates) formed by reactions with amino groups, or obtained by other methods described in the literature, such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbic acid salts, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butates, camphorates, camphorsulfonates, cyclopentylpropionates, digluconates, dodecyl sulfates, ethanesulfonates, formates, transbutenedioates, glucono-p-gluconate, glyceryl phosphates, gluconates, hemisulfates, heptanates, hexanoates, hydroiodates, 2-hydroxy-ethanesulfonates, lacturonates, lactates, laurates, lauryl sulfates, malates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, palmitates, pyruvates, pectates, persulfates, 3-phenylpropionates, picrates, pentanoates, propionates, stearates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, etc. Salts obtained by means of appropriate bases include alkali metals, alkaline earth metals, ammonium, and nitrogen. + Salts of (C1-C4 alkyl)4. This invention also contemplates quaternary ammonium salts formed from any compound containing an N group. Water-soluble or oil-soluble or dispersed products can be obtained by quaternization. Alkali metal or alkaline earth metal salts, including sodium, lithium, potassium, calcium, magnesium, etc. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts, and amine cations resistant to the formation of equilibrium ions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C1-C8 sulfonates, and aromatic sulfonates.

[0153] In this document, the terms “optionally,” “optionally,” or “optionally” generally refer to an event or condition that may, but may not, occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0154] In this document, the terms "optionally substituted" and "substituted or unsubstituted" are used interchangeably. Generally, the term "optionally," whether or not it precedes the term "substituted," indicates that one or more hydrogen atoms in the given structure are substituted by a specific substituent. Unless otherwise indicated, an optional substituent may be substituted at each substituted position of the group. When more than one position in the given structural formula can be substituted by one or more substituents selected from a specific group, the substituents may be substituted at the same or different positions. The substituents mentioned may be, but are not limited to, F, Cl, Br, CN, OH, NH2, NO2, etc.

[0155] In this document, the term "one or more" (e.g., in the definition of substituents in compounds of the general formula of the present invention) means "one, two, three, four or five, especially one, two, three or four, more especially one, two or three, and even more especially one or two".

[0156] Additionally, it should be noted that, unless otherwise explicitly stated, the descriptive terms “each…independently is”, “…each…independently is”, and “…independently is” used in this invention are interchangeable and should be interpreted broadly. They can mean that the specific options expressed by the same symbols in different groups do not affect each other, or that the specific options expressed by the same symbols in the same group do not affect each other.

[0157] In this article, the term "halogen" refers to a fluorine, chlorine, bromine, or iodine atom.

[0158] In this paper, the minimum and maximum carbon atom content in hydrocarbon groups are indicated by prefixes, for example, prefix C. a -C b This refers to a carbon atom containing "a" to "b". For example, "C1-C..." n "C1-C" refers to a saturated / unsaturated carbon chain, either straight or branched, containing 1, 2, 3, 4, 5, ..., or n carbon atoms; further understanding, "C1-C" refers to a saturated / unsaturated carbon chain, either straight or branched. n "Should be interpreted as including any subranges, such as C1-C" 40 C2-C 40 C1-C 18 C3-C 24 C1-C 16 C4-C 10 C4-C8, C1-C3.

[0159] In this article, the term "C1-C" 40 "Alkyl" refers to a saturated monovalent hydrocarbon group with a straight or branched chain having 1, 2, 3, 4, 5, ... or 40 carbon atoms, such as C2-C. 40Alkyl, C2-C 24 Alkyl, C3-C 24 Alkyl, C3-C 11 Alkyl, C4-C 10 Alkyl, C4-C8 alkyl. This includes, but is not limited to, methyl, ethyl, n-propyl (n-Pr, -CH2CH2CH3), isopropyl (i-Pr, -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3)CH2CH3), tert-butyl (t-Bu, -C(CH3)3), and n-pentyl (-CH2CH2CH2CH2C). H3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), n-hexyl (-CH2C) H2CH2CH2CH2CH3), 2-hexyl(-CH(CH3)CH2CH2CH2CH3), 3-hexyl(-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl(-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl(-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl(-CH(CH3)CH2CH(CH3)2), 3 -Methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), n-heptyl, n-octyl, etc., wherein the alkyl group may be independently unsubstituted or substituted by one or more substituents described in this invention.

[0160] In the description of the functional groups of this invention It is used to describe the position of the substituent group.

[0161] In the description of the functional groups of this invention This is used to describe compounds; the carbon-carbon bond here can be a single bond. It can also be a double bond.

[0162] In this article, "ionizable lipids" includes cationic lipids, ionizable lipids, and their derivatives.

[0163] In this article, "liposomes" or "liposome nanoparticles (LNPs)" refers to a drug delivery system that uses biocompatible lipid materials as carriers to dissolve or encapsulate drugs or other bioactive substances in the lipid core or adsorb and attach them to the surface of nanoparticles.

[0164] In this document, the term "pharmaceuticalally acceptable excipient" includes any solvent, dispersion medium, coating material, surfactant, antioxidant, preservative (e.g., antibacterial, antifungal), isotonic agent, salt, pharmaceutical stabilizer, binder, excipient, dispersant, lubricant, sweetener, flavoring agent, colorant, or combination thereof, all of which are known to those skilled in the art (as described in Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329). Except in cases where any conventional carrier is incompatible with the active ingredient, its use in therapeutic or pharmaceutical compositions is covered.

[0165] In this document, the term "treatment" refers to the administration of a drug or compound to an individual to achieve a desired pharmacological and / or physiological effect. This effect may be preventative in terms of complete or partial prevention of a disease or its symptoms, and / or therapeutic in terms of partial or complete cure of a disease and / or adverse effects caused by the disease. As used herein, "treatment" encompasses diseases in mammals, particularly humans, including: (a) prevention of disease or the onset of a condition in an individual who is susceptible but has not yet been diagnosed with the disease; (b) inhibition of disease, such as blocking disease progression; or (c) relief of disease, such as reducing symptoms associated with the disease. As used herein, "treatment" encompasses any administration of a drug or compound to an individual to treat, cure, relieve, improve, reduce, or inhibit the individual's disease, including but not limited to administration of a drug containing a compound described herein to an individual in need.

[0166] The effective amount of the complex or pharmaceutical composition described in this invention can vary depending on the administration method and the severity of the disease to be treated. A preferred effective amount can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to: pharmacokinetic parameters of the bioactive ingredient, such as bioavailability, metabolism, and half-life; the severity of the disease to be treated, the patient's weight, the patient's immune status, and the route of administration. For example, due to the urgency of the treatment condition, several separate doses may be administered daily, or the dose may be reduced proportionally.

[0167] The complexes or pharmaceutical compositions of the present invention can be incorporated into drugs suitable for parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). These drugs can be prepared in various forms, such as liquids, semi-solids, and solid dosage forms, including but not limited to liquid solutions (e.g., injection solutions and infusion solutions) or lyophilized powders. Typical drugs are injection solutions. The aforementioned complexes or pharmaceutical compositions can be administered by intravenous infusion, injection, intramuscular injection, or subcutaneous injection.

[0168] According to embodiments of the present invention, the administration route of the method is intramuscular injection or intravenous injection.

[0169] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0170] The specific compounds of this invention are synthesized using the following general synthetic route:

[0171] When G is -OC (=O)- or -(H)NC (=O)-, the following routes I and II are used for synthesis;

[0172] Route I:

[0173]

[0174] Route II:

[0175]

[0176] Where i is 1 to 4; K is a halogen;

[0177] When G is -C(=O)O- or -C(=O)N(H)-, the following routes III and IV are used for synthesis; Route III:

[0178]

[0179] Route IV:

[0180]

[0181] Where i is 1 to 4; K is a halogen;

[0182] Other compounds included in Formula I of this invention can be prepared using different starting materials via synthetic routes similar to those described above.

[0183] The following detailed examples illustrate the synthesis of compounds with specific structures.

[0184] Example 1: Synthesis of compound 4N4R

[0185] 2-Hexylundecanoic acid 1a (1.2 equiv.) was dissolved in dichloromethane in a round-bottom flask. N,N'-dicyclohexylcarbodiimide (DCC, 2.5 equiv.) and 4-dimethylaminopyridine (DMAP, 0.1 equiv.) were then added, and the mixture was reacted in an ice bath for 1 h. Separately, 6-bromo-1-hexylamine 1b (1.0 equiv.) was dissolved in dichloromethane and added to the same round-bottom flask. The mixture was reacted at room temperature for 6 h. After the reaction was complete, the insoluble solids were removed by filtration, yielding a clear filtrate. The reaction solvent was removed by rotary evaporation under reduced pressure, and the filtrate was purified by silica gel column chromatography to obtain a colorless oily liquid, compound 1. 1,4,7,10-tetraazacyclododecane 1c (1.0 equiv.), sodium hydride (4.0 equiv.), potassium carbonate (6.0 equiv.), and potassium iodide (0.1 equiv.) were added to a pressure-resistant tube. An appropriate amount of acetonitrile was then added, and the mixture was stirred at room temperature for 30 min. Then, compound 1 (6.0 equiv.) and an appropriate amount of acetonitrile (separation will occur), and the mixture was placed in an oil bath at 80°C and reacted for 24 hours. After the reaction was completed, an appropriate amount of dichloromethane was added, and the pale yellow insoluble matter was removed by filtration, yielding a clear, transparent orange-yellow liquid. Rotary evaporation yielded an orange oily liquid, which was the crude product. Separation by silica gel column chromatography yielded a yellow oily product, which was 4N4R.

[0186] The chemical structure and synthetic route of compound 4N4R of this invention are as follows:

[0187]

[0188] Example 2: Synthesis of compound S4N3

[0189] 2-Hexylundecanoic acid 1a (1.2 equiv.) was dissolved in dichloromethane and added to a round-bottom flask. N,N'-dicyclohexylcarbodiimide (DCC, 2.5 equiv.) and 4-dimethylaminopyridine (DMAP, 0.1 equiv.) were then added, and the mixture was reacted in an ice bath for 1 h. Separately, 6-amino-1-hexanol 1b (1.0 equiv.) was dissolved in dichloromethane and added to the same round-bottom flask. The mixture was reacted at room temperature for 6 h. After the reaction was complete, the insoluble solids were removed by filtration, yielding a clear filtrate. The reaction solvent was removed by rotary evaporation under reduced pressure, and the filtrate was purified by silica gel column chromatography to obtain a colorless oily liquid, compound 2. 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (1.0 equiv.), N,N'-dicyclohexylcarbodiimide (DCC, 4.5 equiv.), and 4-dimethylaminopyridine (DMAP, 0.4 equiv.) were added to a pressure-resistant tube, followed by the addition of dichloromethane and reaction in an ice bath for 1 h. Then, compound 2 (6.0 equiv.) was added and the reaction was carried out at room temperature for 24 h. After the reaction was completed, an appropriate amount of dichloromethane was added, and the pale yellow insoluble matter was removed by filtration, yielding a clear, transparent orange-yellow liquid. Rotary evaporation yielded an orange oily liquid, which was the crude product. Separation by silica gel column chromatography yielded a yellow oily product, which was S4N3.

[0190] The chemical structure and synthetic route of compound S4N3 of this invention are as follows:

[0191]

[0192] Example 3: Synthesis of the comparative compound 4N-lipid, where R1, R2, R3, and R4 are all H.

[0193] 1.2 equiv. of 2-hexylundecanoic acid 1a was added to a round-bottom flask and dissolved in dichloromethane. N,N'-dicyclohexylcarbodiimide (DCC, 2.5 equiv.) and 4-dimethylaminopyridine (DMAP, 0.1 equiv.) were then added, and the mixture was reacted in an ice bath for 1 h. Separately, 1.0 equiv. of 6-bromohexanol was dissolved in dichloromethane and added to the same round-bottom flask. The mixture was reacted at room temperature for 6 h. After the reaction was complete, the insoluble solids were removed by filtration, yielding a clear and transparent filtrate. The reaction solvent was removed by rotary evaporation under reduced pressure, and the filtrate was purified by silica gel column chromatography to obtain a colorless oily liquid, compound 3. 1,4,7,10-tetraazacyclododecane (1.0 equiv.), sodium hydride (4.0 equiv.), potassium carbonate (6.0 equiv.), and potassium iodide (0.1 equiv.) were added to a pressure-resistant tube, followed by the addition of an appropriate amount of acetonitrile. The mixture was stirred at room temperature for 30 min. Then, compound 1 (6.0 equiv.) and an appropriate amount of acetonitrile (separation will occur), and the mixture was placed in an oil bath at 80°C and reacted for 24 hours. After the reaction was completed, an appropriate amount of dichloromethane was added, and the pale yellow insoluble matter was removed by filtration, yielding a clear, transparent orange-yellow liquid. Rotary evaporation yielded an orange oily liquid, which was the crude product. Separation by silica gel column chromatography yielded a yellow oily product, which was 4N-lipid.

[0194] The chemical structure and synthetic route of compound 4N-lipid are as follows:

[0195]

[0196] The specific compound structures and their NMR spectra of this invention are shown in Table 1:

[0197] Table 1

[0198]

[0199]

[0200] Example 4: Preparation and formulation performance evaluation of LNPs@mRNA

[0201] The compounds prepared in the above examples were used to construct an mRNA delivery system LNPs@mRNA (LNP loaded with mRNA), and their formulation properties, such as particle size, potential, encapsulation efficiency, and mRNA integrity, were investigated to evaluate their formulation characteristics.

[0202] The preparation method of LNPs@mRNA is as follows:

[0203] (1) Solution preparation: Ionizable lipids (the compounds prepared in this invention), DOPE, cholesterol (Chol), and DMG-PEG2000 were dissolved in anhydrous ethanol to obtain lipid solutions. Meanwhile, Fluc mRNA was dissolved in 50 mM PBS buffer solution (prepared with RNase-free water) and finally prepared into 10 mM PBS buffer solution as the aqueous phase, which was then diluted to a suitable concentration for later use.

[0204] (2) LNP preparation: The lipid solution obtained in step (1) was mixed with the mRNA solution to obtain the initial LNP formulation. The mass ratio of ionizable lipids to mRNA was controlled at 20:1. The mixing process was carried out in a microfluidic device with the following microfluidic process parameters: the volume ratio of ethanol phase to water phase was 1:4, and the total flow rate was 12 mL / min.

[0205] (3) Ultrafiltration: The initial LNP preparation was diluted 25 times with PBS buffer and ultrafiltered to the initial volume to obtain the final LNP preparation. The ultrafiltration process removed ethanol from the initial preparation, yielding the LNPs@mRNA drug of the compound of this invention. Ultrafiltration process parameters: membrane pore size 100kDa, ultrafiltration pressure 0.2MPa, rotation speed 100-200rpm.

[0206] LNPs@mRNA formulation properties investigation:

[0207] Particle size potential determination: The particle size potential of the LNPs@Fluc compounds of this invention was determined using a Malvern laser particle size analyzer. A certain volume of freshly prepared LNPs@Fluc solution was diluted 10 times with pure water, and its particle size and potential were determined using a Malvern laser particle size analyzer. Each sample was measured in triplicate at a temperature of 25°C. The particle size potential determination results are as follows: Figure 1 As shown.

[0208] Encapsulation efficiency testing: Quant-iT TM RiboGreen TM The reagent kit is used to test the encapsulation rate.

[0209] The mRNA encapsulation efficiencies of the 4N4R@mRNA and S4N3@mRNA formulations were 96.2% and 92.3%, respectively.

[0210] mRNA stability study: 4N4R@mRNA was stored at -80℃, -20℃, and 4℃ for 1, 8, 13, and 36 days, respectively. Its particle size, PDI, and mRNA content were then assessed. In vivo expression was also evaluated after 7, 14, and 36 days of storage to assess the potential for vaccine development using this compound. The results of the 4N4R@mRNA stability study are as follows: Figure 2 and Figure 3 As shown, Figure 2 As shown, the particle size of the 4N4R@mRNA of the present invention did not change significantly at -80℃; as Figure 3 As shown, the expression level of the 4N4R@mRNA of the present invention remained unchanged in mice at -80°C.

[0211] This indicates that the 4N4R@mRNA of the present invention can be stably stored for 36 days at -80°C.

[0212] Based on the above results, the compounds of the present invention exhibit excellent performance when prepared into LNPs formulations, and can be further used in the development of nano-formulations.

[0213] Example 5: Investigation of the mRNA delivery performance of the compounds of the present invention

[0214] The expression of green fluorescent protein in HEK293T cells was detected using the 4N4R@mRNA vaccine prepared from the compounds obtained in the above embodiments of the present invention. The procedure was as follows:

[0215] (1) Cell plating: Adjust the cell suspension to the appropriate concentration with 1640 complete culture medium, and plate the cells in 24-well plates with 5×10⁶ cells per well. 5 / 500μL / well, and continue to incubate in an incubator at 37℃ and 5% CO2 for 24h.

[0216] (2) Preparation of formulation and cell transfection: 4N4R@mRNA and positive control formulation lipo 2000@mRNA were prepared according to the method of the above embodiment. The mRNA concentrations of the administration formulations were controlled to be 0.25, 0.5 and 1 μg, respectively. 20 μL was administered to each well and incubated in a 37°C, 5% CO2 incubator for 24 h.

[0217] The delivery efficiency of the compound 4N4R@mRNA of this invention is as follows: Figure 4 As shown, the protein expression level of the example is comparable to that of the positive control preparation, and it has good transfection efficiency.

[0218] Example 6: Investigation of in vivo expression of LNPs prepared from the compounds of the present invention

[0219] The expression of LNPs prepared from the 4N4R compound and the 4N-lipid compound of this invention in animals was investigated by intravenous injection.

[0220] The LNP@mRNA formulation was prepared according to the method described in Example 4. The mRNA concentration of the formulation was adjusted to 0.05 mg / mL with PBS solution, and the osmotic pressure of the formulation was adjusted to isotonicity. 200 μL was injected intravenously into animals (3 animals per group), with PBS used as a negative control. Animals maintained a normal diet after administration. Six hours after administration, 200 μL of substrate solution (15 mg / mL, potassium fluorescein) was injected intraperitoneally. Timing began after substrate injection, and euthanasia was performed 10 minutes later. The heart, liver, spleen, lungs, and kidneys were rapidly dissected, and the bioluminescence intensity of each isolated organ was measured using an IVIS instrument with an exposure time of 60 s. The total flux of each organ was calculated after imaging.

[0221] Experimental results are as follows Figure 5 As shown, the ionizable lipid compound 4N4R of the present invention can target and deliver mRNA to mice, especially the spleen, after encapsulation of mRNA. Compared with compound 4N-lipid, the expression level of mRNA encapsulated by 4N4R in the spleen is higher and significantly different.

[0222] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0223] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt or deuterated compound thereof: in, X is any integer from 0 to 3; Y is any integer from 1 to 3; A is a substituted or unsubstituted C0-C3 alkylene group; B is a substituted or unsubstituted C0-C5 alkylene group; D is a substituted or unsubstituted C1-C5 alkylene group; E represents substituted or unsubstituted C1-C. 20 Straight-chain or branched alkyl groups; M is -O-, -S-, or -NH-; G is -OC(=O)-, -C(=O)O-, -C(=O)N(H)- or -(H)NC(=O)-; R1, R2, R3, and R4 are each independently selected from H, -OH, -SH, =O, and -NR5R6, wherein at least two of R1, R2, R3, and R4 are not H; R5 and R6 are each independently selected from H, -OH, -SH, and C1-C3 alkyl groups.

2. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, When X is 1, 2, or 3; when two of R1, R2, R3, or R4 are H, it means that two adjacent numbers are not both H at the same time.

3. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, Y is 1 or 2.

4. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, M is -O-.

5. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, R1, R2, R3, and R4 are -OH.

6. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, X is 1; Y is 1; A is a non-substituted C0-C3 alkylene group; B is a non-substituted C0-C5 alkylene group; D is a non-substituted C1-C3 alkylene group; M is O; R1, R2, R3, and R4 are each independently selected from OH and -SH.

7. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, B is a C0 alkylene group; R1, R2, R3, and R4 are equal to 0.

8. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, E represents substituted or unsubstituted C5-C. 18 Straight-chain or branched alkyl groups. Furthermore, E is selected from the following groups:

9. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, X is 1; Y is 1 or 2 A is -CH2-; B is -CH2-; D is -CH2- or -CH2CH2-; M is O; R1, R2, R3, and R4 are OH; G is -OC(=O)-, -C(=O)O-; E is 10. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, It has the following structure:

11. A composition comprising the compound of any one of claims 1 to 10 or a pharmaceutically acceptable salt or deuterated compound thereof, characterized in that, in, The composition also includes pharmaceutically active molecules and pharmaceutically acceptable excipients; Furthermore, the active pharmaceutical molecule is selected from DNA, ASO, siRNA, miRNA, mRNA, ribozymes, nucleic acid aptamers, or combinations thereof; The pharmaceutical composition contains at least one excipient selected from neutral phospholipids, steroids, and polyethylene glycol lipids.

12. The pharmaceutical composition according to claim 11, characterized in that, The compound constitutes 30% to 60% of the composition in molar proportion; Further, the pharmaceutical composition further comprises: (i) 5-25% molar neutral lipids, (ii) 25-55% molar steroids or steroid analogs, and (iii) 0.5-10% molar PEG-lipids. Further, the pharmaceutical composition is prepared into lipid nanoparticles.

13. Use of the pharmaceutical composition according to claim 11 in the preparation of liposomes, lipid nanoparticles, drug carriers or complexes.

14. A method for treating a disease using the compound of any one of claims 1 to 10 or a pharmaceutically acceptable salt or deuterated compound thereof, the method comprising loading the compound or a pharmaceutically acceptable salt or deuterated compound thereof with a drug and delivering the drug into cells.

15. Use of the pharmaceutical composition of claim 11 in the preparation of a medicament for treating a disease.

16. A method of treating a disease with the pharmaceutical composition of claim 12, the method comprising loading the lipid nanoparticles with the drug and delivering the drug to the spleen.

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