Ionized lipids and lipid nanoparticles containing the same

Abstract

The present invention provides an ionizable lipid of formula (I) or a pharmaceutically acceptable salt thereof or any one of its stereoisomers, lipid nanoparticles comprising the ionizable lipid, particularly as an encapsulating agent, optionally containing a pharmaceutically active agent, and pharmaceutical compositions comprising the lipid nanoparticles. The present invention also provides lipid nanoparticles for use in medicine, pharmaceutical compositions comprising the lipid nanoparticles, and uses of the lipid nanoparticles as an encapsulating agent. [Formula 1] TIFF2025538495000271.tif29159

Description

[Technical Field]

[0001] This application claims the benefit of European Patent Application No. 22383119.9 filed November 21, 2022, European Patent Application No. 23382067.9 filed January 27, 2023, and European Patent Application No. 23382401.0 filed April 28, 2023.

[0002] The present disclosure relates to ionizable lipids and lipid nanoparticles (LNPs) comprising said ionizable lipids. These LNPs can be used as non-viral vectors for the delivery of active ingredients, including polynucleotides, to cells. [Background technology]

[0003] The treatment of various diseases, such as infectious diseases, is changing rapidly today, especially since new polynucleotide delivery systems have been designed and their efficiency optimized to provide useful alternatives to conventional treatments.

[0004] Gene delivery systems intended to contain encapsulated polynucleotides such as RNA for systemic delivery must be safe and non-toxic, nanometer-scale, provide protection for the polynucleotides against degradation, remain intact within the system for a period of time to reach their target, and be readily degraded once the payload is released.

[0005] The use of RNA in gene therapy and vaccination is generally considered safer than the use of DNA because RNA does not stably integrate into the genome of transfected cells. Furthermore, RNA degrades more readily in vivo, reducing the risk of developing unwanted anti-RNA antibodies, which can reduce therapeutic efficacy and cause severe side effects. Two major limitations of RNA-based therapeutics are low transfection rates and limited protein production efficiency. To compensate for these, the dosage must be increased to achieve the desired therapeutic effect. Therefore, increasing the dosage also increases costs and can result in some unwanted side effects.

[0006] One of the more powerful intracellular delivery technologies for encapsulating and delivering active molecules, such as genetic material (e.g., mRNA) in vaccines, is lipid nanoparticles (LNPs). Currently, LNPs used in commercial mRNA vaccines typically contain four types of lipids: cationic or ionizable lipids, structural lipids (e.g., sterols such as cholesterol), PEG-modified lipids, and non-cationic lipids such as phospholipids. Ionizable lipids are particularly important in LNPs.

[0007] Ionizable lipids have a significant impact on the protection of genetic material encapsulated within LNPs, as the properties of ionizable lipids enable LNPs to maintain the structural and physicochemical properties of the genetic material until it reaches the target (e.g., tissue, cell) where it is to be released.

[0008] Ionizable lipids typically have the following general structure: Lipophilic part---Binding group---Crosslinking---Hydrophilic part Shows.

[0009] The state of the art presents many examples in which the hydrophilic moiety contains an ionizable amine (a functional group whose formal charge is affected by changes in pH). It has also been disclosed that the presence of an ester group, which can be easily hydrolyzed by enzymes, facilitates the degradation of the ionized lipid once the genetic material is released into the tissue / cell of interest, thereby improving its biocompatibility and biodegradability. The proximity of the ester group to the hydrophilic moiety is also said to have a significant impact on the efficacy of the lipid.

[0010] As an example, Moderna uses the SM-102 lipid in its SARS-CoV-2 vaccine formulation (Spikevax), and Pfizer has licensed ALC-0315 from Acuitas for use in its vaccine (Comirnaty). [ka]

[0011] Molla MR, et al. (see Molla MR, et al. "One-Pot Parallel Synthesis of Lipid Library via Thiolactone Ring Opening and Screening for Gene Delivery", Bioconjug. Chem. 2018, vol. 29(4), pp. 992-999. doi:10.1021 / acs.bioconjchem.8b00007) disclose a combinatorial library of lipidoids containing reducible disulfide groups and having hydrophobic tails that allow stable liposomes to be obtained.

[0012] Despite the continued improvement of such LNP delivery systems, efficient and specific delivery of targeted drugs remains problematic. Thus, there remains a need for alternative ionizable lipids and / or LNPs that overcome some of the drawbacks of the prior art and, in particular, exhibit good stability, high transfection efficiency, and safety. Summary of the Invention

[0013] The present inventors have developed new ionizable lipids that allow for the preparation of lipid nanoparticles (LNPs) that can be effectively used as non-viral vectors for the delivery of active ingredients, including polynucleotides, to cells. In particular, LNPs containing the ionizable lipids of the present disclosure exhibit enhanced / improved transfection rates.

[0014] The inventors have discovered an ionizable lipid of formula (I) as defined herein below, which comprises a polar head (R3), at least one thioether moiety, at least one asymmetric center, at least one ester moiety, and two moieties A and B selected from amides or esters: [ka] wherein A, B, X, Z, R3, m, n, p, q, and t are as defined in this disclosure. are particularly useful in preparing LNPs that can encapsulate active agents. Furthermore, the inventors have found that the in vivo transfection rate of LNPs prepared with the ionizable lipids of the present disclosure and containing active agents is unexpectedly high compared to other commercially available or standard compositions known in the art.

[0015] Thus, it is clear from the data provided in the Examples that the ionizable lipids of the present disclosure provide a new means to overcome some of the limitations of known LNPs.

[0016] The higher transfection efficiency of the disclosed LNPs may allow for reduced therapeutic doses of polynucleotides, such as RNA, required in gene therapy and vaccination, which may also help reduce associated secondary effects. At the same time, the disclosed LNPs exhibit favorable thermal and mechanical properties, demonstrating that their transfection efficiency in vivo is maintained for 3 months even after storage at 4°C without further precautions. This represents a long shelf life under normal refrigeration conditions, and this shelf life could be extended at lower temperatures (e.g., at home freezer temperatures of -25°C to -15°C, or even at temperatures as low as -90°C).

[0017] Thus, a first aspect of the present disclosure provides an ionizable lipid of formula (I): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof, During the ceremony, A is selected from the group consisting of -NR'-C(O)-, -C(O)-NR'-, -C(O)-O-, and -OC(O)-, where R' is selected from the group consisting of H, methyl, and ethyl; B is selected from the group consisting of -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -OC(O)-; n, m, and p are independently selected from 0, 1, 2, 3, 4, 5, and 6; q is selected from 1, 2 and 3; t is selected from 0, 1 and 2; and where q+t=1, 2 or 3, R3 is a heterocycle containing at least one N atom, or alternatively R3 is [ka] and wherein Ra and Rb are independently a linear or branched C1-C6 alkyl optionally substituted with a hydroxyl group; X is -D-R2, [ka] is selected from wherein j, j', and j'' are independently selected from 0, 1, and 2; Z is -R1 and [ka] is selected from wherein f and f' are independently selected from 0, 1, 2, 3, 4, 5, and 6; g and g' are independently selected from 1, 2, 3, 4, 5 and 6; each D is independently selected from -C(O)-O- or -OC(O)-; and R1, R'1, R2, R'2, and R''2 are independently a straight or branched chain C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R1, R'1, R2, R'2, and R''2 are optionally substituted with one or more substituents selected from the group consisting of -OH, -COOR4, and -C(=O)SR4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl; however, When q+t=1, X=-D-R2, D=-OC(O)-, A=-NH-C(O)-, B=-NH-C(O)-, Z=R1, and R1 is a linear or branched alkyl, the moiety -(CH2) m -R1 contains at least 7 carbon atoms, The following compound: S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-methyl homocysteate: [ka] and S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-L-cysteic acid methyl ester [ka] (Both are disclosed in Churusova, S. et al. (2021). "Palladium(II) Pincer Complexes of Functionalized Amides with S-Modified Cysteine ​​and Homocysteine ​​Residues: Cytotoxic Activity and Different Aspects of Their Biological Effect on Living Cells." Inorganic Chemistry. 2021, vol. 60, pp. 9880-9898.) and the compound (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole disclosed in U.S. Pat. No. 4,929,736. is excluded.

[0018] Another aspect of the invention relates to LNPs comprising ionizable lipids of formula (I) as defined herein, including ionizable lipids not claimed above or below.

[0019] In another embodiment, the LNPs of the present disclosure can further comprise a pharmaceutically active agent and thus can be formulated into a pharmaceutical product using excipients and carriers.

[0020] Accordingly, another aspect of the present invention relates to pharmaceutical compositions comprising the present LNPs, which comprise a pharmaceutically active agent, as defined herein, and a pharmaceutically acceptable excipient or carrier.

[0021] LNPs or pharmaceutical compositions of the present disclosure that include a pharmaceutically active agent can be used for pharmaceutical applications.

[0022] Thus, another aspect of the present invention relates to an LNP or pharmaceutical composition of the present disclosure comprising a pharmaceutically active agent for use in medicine, particularly for use in a method for treating a disease or disorder in a subject in need thereof, or for use in a method for inducing an immune response in a subject, for use in a method for therapeutic immunization of a subject, for use as a vaccine, or for use in gene therapy, which method comprises administering a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition to a subject.

[0023] Another aspect of the present invention relates to the use of LNPs as defined herein as encapsulating agents for active ingredients. [Brief explanation of the drawings]

[0024] [Figure 1] Referring to Example 7, protein expression is shown in mice administered LNPs prepared with the following lipids of the present invention containing RNA as the active ingredient: VC-LC-0163, VC-LC-0289, VC-LC-0374, VC-LC-0389, VC-LC-0553, VC-LC-0554, VC-LC-0353, and VC-LC-0355. The lipids tested and illustrated contain the same R3 substituent (polar head) in all but one case and different combinations of R2 and R3 substituents, and show successful transfection results, as demonstrated by the images and total flux measurements for each lipid in Table 7 (A-C). [Figure 2] Regarding Comparative Example 9, in vivo protein expression is shown in mice injected with LNPs containing VC-LC-0236 as the ionizable lipid in the LNP composition (FIG. 2A) and VC-LC-0588 as the ionizable lipid in the LNP composition (FIG. 2B). [Figure 3] Referring to Example 10, protein expression in mice injected with LNPs from Table 11 having different composition ratios of the ionizable lipid VC-LC-0431, DOPE, cholesterol, and DMG-PEG2000 is shown. DETAILED DESCRIPTION OF THE INVENTION

[0025] All terms used herein in this application shall be understood to have their ordinary meaning known in the art unless otherwise specified. Other, more specific definitions for certain terms used in this application are set forth below, and are intended to be applied uniformly throughout this specification and claims, unless a specifically and expressly set forth definition gives a broader definition.

[0026] As used herein, the indefinite articles "a" and "an" are synonymous with "at least one" or "one or more." Unless otherwise specified, definite articles used herein, such as "the," also include plural nouns.

[0027] The term "and / or" means that any one of the alternatives to which it relates is possible, or that at least two of the alternatives may occur simultaneously.

[0028] The term "moiety" refers to a specific segment or functional group of a molecule or compound.

[0029] As used herein, the term "subject" refers to any mammal, including both human and non-human mammals.

[0030] The term "C1-C# alkyl" as used herein refers to a saturated, straight or branched chain hydrocarbon containing 1 to # carbon atoms and which is optionally substituted. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, neopentyl, n-hexyl, decyl, isodecyl, undecyl, dodecyl, tetradecyl, and hexadecyl.

[0031] The term "C2-C# alkenyl" refers to an optionally substituted, unsaturated, straight or branched hydrocarbon chain containing 2 to # carbon atoms and at least one double bond. Examples of alkenyl groups include, but are not limited to, ethenyl (i.e., vinyl), allyl, propenyl, butenyl, pentenyl, hexenyl, dodecenyl, tetradecenyl, and hexadecenyl.

[0032] The term "C2-C# alkynyl" refers to an optionally substituted, unsaturated, straight or branched hydrocarbon chain containing 2 to # carbon atoms and at least one triple bond. Examples of alkynyl groups include, but are not limited to, ethynyl, prop-1-ynyl, prop-2-ynyl, 1-methylprop-2-ynyl, but-1-ynyl, but-2-ynyl, and but-3-ynyl.

[0033] The term "heterocycle containing at least one N atom" refers to an optionally mono- or polysubstituted ring system containing one or more rings, wherein at least one ring contains at least one nitrogen.

[0034] The term "optionally substituted" means that the number of substituents can be equal to or different from zero. Unless otherwise specified, an optionally substituted group may be substituted with as many optional substituents as can be accommodated by replacing a hydrogen atom on any available carbon or nitrogen atom with a non-hydrogen substituent.

[0035] The term "polynucleotide" is used interchangeably with "nucleic acid" and refers to a polymer of nucleotides, either ribonucleotides or deoxyribonucleotides. Polynucleotides formed from ribonucleotides may be referred to as "RNA polynucleotides," "ribonucleic acid," or simply "RNA," while polynucleotides formed from deoxyribonucleotides may be referred to as "DNA polynucleotides," "deoxyribonucleic acid," or simply "DNA." Polynucleotides may be single- or double-stranded, and optionally incorporate synthetic, non-natural, or modified nucleotides that can be incorporated into DNA or RNA. An "artificial polynucleotide" refers to a polynucleotide having a sequence that does not occur in nature or that has been altered by human intervention.

[0036] As used herein, the term "messenger RNA," abbreviated as "mRNA," refers to any RNA polynucleotide that encodes a polypeptide of interest and that can be translated in vitro, in vivo, or ex vivo to produce the encoded polypeptide of interest. Typically, mRNA is single-stranded and contains an ORF in its structure.

[0037] As used herein, "open reading frame" or "ORF" refers to a sequence of several nucleotide triplets that encodes a polypeptide, i.e., that can be translated into a polypeptide sequence.

[0038] As used herein, "DNA construct" refers to an artificial polynucleotide comprising a sequence of interest operably linked to an expression promoter, which controls the expression of the sequence of interest.

[0039] As used herein, "expression vector" refers to a vector used to introduce a particular nucleic acid, typically a DNA construct, into a target cell for expression of the nucleic acid by the cell. Examples of suitable expression promoters and expression vectors include those conventionally used in molecular biology and known to those skilled in the art.

[0040] The term "polypeptide" refers to any peptide or protein containing two or more amino acids joined together by peptide bonds or modified peptide bonds (i.e., peptide isosteres). "Polypeptide" refers to both short chains, commonly referred to as peptides, oligopeptides, or oligomers, and to longer chains, commonly referred to as proteins.

[0041] The expression "therapeutically effective amount" as used herein refers to an amount of a compound that, when administered, is sufficient to prevent or alleviate to some extent the occurrence of one or more symptoms of the disease being treated. The specific dose of a compound administered according to the present invention will naturally be determined by the specific circumstances surrounding the case, including the compound administered, the route of administration, the specific condition being treated, the specific circumstances of the individual subject being treated, and similar considerations. The term "pharmaceutical product" also encompasses the concept of "veterinary composition". Thus, the term relates to a composition that is therapeutically effective when administered by any desired or applicable route to any animal, including humans.

[0042] According to the present invention, the term "antigen" is a compound that can be recognized by the immunoglobulin receptor of a B cell or a T cell receptor when complexed with an MHC. Preferably, an "antigen" is a polypeptide.

[0043] As used herein, the term "nanoparticle" refers to a particle having at least two dimensions on the nanometer scale, particularly all three dimensions on the nanometer scale, where the nanoscale ranges from about 1 nm to about 500 nm. "Nanoparticle" refers to a particle having at least two dimensions on the nanometer scale, particularly when the nanoparticle is substantially rod-shaped with a substantially circular cross-section, such as a nanowire or nanotube, where these two dimensions are the cross-section of the nanoparticle. As used herein in the context of nanoparticle compositions, "size" or "average size" refers to the average diameter of the nanoparticle composition. As used herein, the term "lipid nanoparticle" refers to a nanoparticle whose outer envelope is made entirely or partially of lipid.

[0044] As used herein, "polydispersity index (PDI)" is a ratio that describes the homogeneity of the particle size distribution of a system. A small value, e.g., less than 0.3, indicates a narrow particle size distribution.

[0045] The term "zeta potential" used herein refers to the electrokinetic potential of lipids, for example, in particle compositions.Zeta potential is also defined as the potential difference between the dispersion medium and the fixed layer of fluid attached to the dispersed particle.This is generally accepted as a quantification of the magnitude of charge, and is often the only method available for characterizing bilayer properties.

[0046] As used herein, "apparent pKa" refers to an experimentally determined value obtained from the average ratio of all ionized and deionized groups in a nanoparticle. The apparent pKa is different from the intrinsic pKa of any individual molecule and is an important parameter for the performance of nanoparticles to encapsulate RNA. The apparent pKa of a nanoparticle can be measured by various techniques known in the art. For example, acid-base titration and 2-(p-toluidino)-6-naphthalenesulfonic acid (TNS) fluorescence are widely used in the art. Nanoparticles with an optimal pKa have negligible charge at physiological pH, preventing nonspecific binding and toxicity in the body. The optimal pKa of a nanoparticle plays an important role in the endosomal escape mechanism and release of RNA into the cytosol for therapeutic effect.

[0047] Ionized lipids As mentioned above, the first aspect of the present invention provides an ionizable lipid of formula (I): [ka] or a pharmaceutically acceptable salt thereof, or any one stereoisomer thereof, wherein A, B, m, n, p, q, t, R3, X and Z are as defined above.

[0048] In one embodiment of the ionizable lipid, n=t=m=0, q=p=1, X is [ka] wherein one of j and j′ is 0 and the other is 1; When D is -OC(O)-, A=-C(O)-NH-, B=-NH-C(O)-, Z is R1, and R1 is C1-C18 alkyl, R2 and R'2 are different from C7-C18 alkyl.

[0049] In one embodiment, the ionizable lipid is a compound of formula (II): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A, B, D, n, m, p, q, t, q+t, R1 and R2 are as defined above, however, When q+t=1, X=-D-R2, D=-OC(O)-, A=-NH-C(O)-, B=-NH-C(O)-, Z=R1, and R1 is a linear or branched alkyl, the moiety -(CH2) m -R1 contains at least 7 carbon atoms, The following compound: S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-methyl homocysteate: [ka] S-(2-Methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-L-cysteic acid methyl ester [ka] and (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole are excluded.

[0050] In another embodiment of the ionizable lipid of the present disclosure, A is —NH—C(O)— or —C(O)—NH—, i.e., the ionizable lipid is an ionizable lipid of formula (II): [ka] Formula (II) or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A is [ka] and B is [ka] and D is [ka] and n, m, and p are independently 0, 1, 2, 3, 4, 5, or 6; q is selected from 1, 2 and 3; t is selected from 0, 1 and 2; and where q+t=1, 2 or 3, R1 and R2 are independently a linear or branched C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R1 and R2 are optionally substituted with one or more substituents selected from the group consisting of -OH, -COOR4, and -C(=O)S-R4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl; and R3 is a heterocycle containing at least one N atom, or alternatively R3 is [ka] wherein Ra and Rb are independently a linear or branched C1-C6 alkyl optionally substituted with a hydroxyl group; however, When q+t=1, X=-D-R2, D=-OC(O)-, A=-NH-C(O)-, B=-NH-C(O)-, Z=R1, and R1 is a linear or branched alkyl, the moiety -(CH2) m -R1 contains at least 7 carbon atoms, and The following compound: S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-methyl homocysteate: [ka] S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-L-cysteic acid methyl ester: [ka] and (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole are excluded.

[0051] As used herein, the term "pharmaceutically acceptable salts" includes any salt formed from a pharmaceutically acceptable non-toxic acid, including inorganic or organic acids. There are no limitations on the salt, other than that it must be pharmaceutically acceptable when used for therapeutic purposes. Pharmaceutically acceptable salts of the ionizable lipids of the present disclosure can be prepared by methods known in the art. For example, they can be prepared from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts are prepared by, for example, reacting the free acid or base form of the ionizable lipids of the present disclosure with a stoichiometric amount of an appropriate pharmaceutically acceptable base or acid in water or an organic solvent, or a mixture thereof.

[0052] Examples of pharmaceutically acceptable salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid, and organic acids such as succinic acid, maleic acid, acetic acid, fumaric acid, citric acid, tartaric acid, benzoic acid, malic acid, lactic acid, formic acid, propionic acid, glycolic acid, camphorsulfuric acid, mandelic acid, benzenesulfonic acid, p-toluenesulfonic acid, oxalic acid, methanesulfonic acid or naphthalenesulfonic acid, as well as base addition salts formed with organic bases such as alkali metals and alkaline earth metals, N,N-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, lysine and procaine. The ionizable lipids of the present disclosure and their salts may differ in some physical properties, but for the purposes of the present invention, they are equivalent.

[0053] The ionizable lipid of the present disclosure has asymmetric center, and therefore can produce various stereoisomers.The term "stereoisomer" used herein refers to all isomers of individual ionizable lipids that differ only in the orientation of their atoms in space.The term stereoisomer includes enantiomers, racemates, racemic mixtures, geometric isomers (cis / trans or syn / anti or E / Z) and diastereomers.The present invention relates to each of these stereoisomers and also to their mixtures.

[0054] The preparation processes described herein can be modified to obtain enantiomerically pure compounds as well as mixtures of stereoisomers. Specific stereoisomers or mixtures can be prepared by a variety of processes, including the use of stereospecific reagents, or by introducing chiral centers into the compounds during the preparation process. Furthermore, once the compounds are prepared, stereoisomers can be separated by standard resolution techniques known to those skilled in the art.

[0055] In all embodiments of the present invention that refer to the ionizable lipids of the present disclosure, a pharmaceutically acceptable salt or stereoisomer of the ionizable lipid or a pharmaceutically acceptable salt thereof is always envisioned, even if not specifically mentioned.

[0056] The ionizable lipids of the present disclosure also include isotopes of the illustrated structures. "Isotopes" refer to atoms with the same atomic number but different mass numbers resulting from different numbers of neutrons in their nuclei. For example, isotopes include, but are not limited to, tritium, deuterium, 13 C or 14 C or 15 N. Additionally, the compounds or salts of the present disclosure can be prepared in combination with solvents or water molecules by routine methods to form solvates and hydrates.

[0057] The ionizable lipids of the present disclosure are characterized by their retention time, mass spectrometry, particle size distribution, polydispersity index and zeta potential.These parameters can be measured by methods well known in the art.Some of them are shown in more detail in the following examples.

[0058] According to one embodiment of formula (II), n is selected from 0, 1, 2 and 3; p is selected from 0, 1, 2 and 3; and m is selected from 0, 1, 2 and 3.

[0059] Another embodiment is an ionizable lipid of formula (II): [ka] or a pharmaceutically acceptable salt thereof, or any one stereoisomer thereof; During the ceremony, A is [ka] and B is [ka] and D is [ka] and n, m, and p are independently 0, 1, 2, 3, 4, 5, or 6; q is selected from 1 and 2; t is selected from 0 and 1; and here q+t=2, R1 and R2 are independently a linear or branched C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R1 and R2 are optionally substituted with one or more substituents selected from the group consisting of -OH, -COOR4, and -C(=O)S-R4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl; and R3 is a heterocycle containing at least one N atom, or alternatively R3 is [ka] wherein Ra and Rb are independently a straight or branched C1-C6 alkyl optionally substituted with a hydroxyl group.

[0060] According to another embodiment, optionally in combination with one or more features of the various embodiments described above, R3 is a 5- or 6-membered ring containing one N atom and optionally a second heteroatom, preferably selected from N and O.

[0061] In another embodiment, in any combination with one or more features of the various embodiments described above, R3 has the following structure: [ka] is selected from.

[0062] In another embodiment, in any combination with one or more features of the various embodiments described above, n is selected from 0, 1, 2 or 3; p is selected from 0, 1, 2 or 3; m is selected from 0, 1, 2 or 3; and optionally t is 0 or 1, q is 1 or 2, and t+q=2.

[0063] In another embodiment, n is 2 or 3, m=t=0, p=q=2, A=B= [ka] and During the ceremony, R1 is a C10-C20 branched alkyl; R2 is a C2-C30 linear or branched alkyl or a C6-C24 linear alkenyl or alkynyl; and R3 is [ka] and In the formula, Ra=Rb=C1-C4 alkyl.

[0064] In another embodiment, n=2, and R2 is a C2-C30 straight or branched alkyl, and R3 is [ka] and In the formula, Ra and Rb are C1 to C4 alcohol terminal alkyls.

[0065] In another embodiment, n is 2 or 3, m=t=0, p=q=2, A is [ka] and R1 is a C10 to C20 branched alkyl, R2 is a C3 to C30 linear or branched alkyl, and R3 is an imidazole ring.

[0066] In another embodiment, n=3, m=t=0, p=q=2, A is [ka] and R1 is a C10 to C20 branched alkyl, R2 is a C4 to C20 linear or branched alkyl, and R3 is a pyrrolidine ring.

[0067] In another embodiment, n=2, m=t=0, p=q=2, A is [ka] and R1 is a C10 to C20 linear or branched alkyl, R2 is a C4 to C20 linear or branched alkyl, and R3 is N-4-methylpiperazine.

[0068] In another embodiment, n is selected from 2 or 3; m=t=0 and p=q=2, A=B= [ka] and R1 is a C10 to C22 linear or branched alkyl, alkenyl, or alkynyl, R2 is a C3 to C30 linear or branched alkyl or a C3 to C30 linear alkenyl or alkynyl, and R3 is a morpholino ring.

[0069] In another embodiment, n is selected from 2 or 3, m=0, p=2, q=t=1; A is [ka] and B=D, R1 is a C10-C22 straight or branched alkyl, R2 is a C3-C30 straight or branched alkyl or a C3-C30 straight alkenyl or alkynyl, and R3 is a morpholino ring.

[0070] In another embodiment, n=q=p=2, t=m=0, A is [ka] and B is [ka] and D is [ka] and R1 is a C10-C22 branched alkyl; R2 is a C8-C20 straight chain alkenyl or alkynyl or a C3-C28 straight chain alkyl optionally substituted with one or more substituents selected from OH, -COOR4 and -COSR4, wherein R4 is a C1-C6 alkyl, a C2-C6 alkenyl, a C2-C6 alkynyl or a C12-C25 branched chain alkyl; and R3 is -N(CH3)2 or [ka] is.

[0071] In another embodiment, the ionizable lipid of the present disclosure is a compound of formula (III): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A is —NH—C(O)— or —C(O)—NH—; R1, R2, and R'2 are independently selected from linear or branched C1-C30 alkyl, C2-C30 alkenyl, and C2-C30 alkynyl, wherein each of R1, R2, and R'2 is optionally substituted with one or more substituents selected from the group consisting of -OH, -COO(R4), and -COSR4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl; and B, D, n, m, p, q, t, q+t, j, j′ and R3 are as defined above.

[0072] In certain embodiments of compounds of Formula (III), when n=t=m=0, q=p=1, A=B=—NH—C(O)—, one of j and j′ is 0 and the other is 1, D=—OC(O)—, and R1 is C1-C18 alkyl, then R2 and R′2 are not C7-C18 alkyl.

[0073] In another embodiment, the ionizable lipid of the present disclosure is a compound of formula (IV): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A is —NH—C(O)— or —C(O)—NH—; B, D, n, m, p, q, t, q+t, f, f', g, g' and R3 are as defined above.

[0074] Examples of ionizable lipids of formula (I), formula (II), formula (III) and formula (IV) include the compounds shown in Table 1 below. [Table 1] JPEG2025538495000043.jpg230136JPEG2025538495000044.jpg235139JPEG2025538495000045.jpg239159JPEG2025538495000046.jpg250159JPEG2025538495000047.jpg249159JPEG2025538495000048.jpg252159JPEG2025538495000049.jpg238136JPEG2025538495000050.jpg243159JPEG2025538495000051.jpg236159JPEG2025538495000052.jpg235159TIFF2025538495000053.tif242158TIFF2025538495000054.tif242149JPEG2025538495000055.jpg253159TIFF2025538495000056.tif242155JPEG2025538495000057.jpg238159JPEG2025538495000058.jpg253159TIFF2025538495000059.tif242158TIFF2025538495000060.tif242154TIFF2025538495000061.tif242148JPEG2025538495000062.jpg237159TIFF2025538495000063.tif242147JPEG2025538495000064.jpg218140JPEG2025538495000065.jpg243159TIFF2025538495000066.tif242158TIFF2025538495000067.tif242150TIFF2025538495000068.tif242154TIFF2025538495000069.tif242156TIFF2025538495000070.tif242153TIFF2025538495000071.tif242155TIFF2025538495000072.tif242154TIFF2025538495000073.tif242148TIFF2025538495000074.tif237159JPEG2025538495000075.jpg235159TIFF2025538495000076.tif242157JPEG2025538495000077.jpg244159TIFF2025538495000078.tif242154TIFF2025538495000079.tif242155JPEG2025538495000080.jpg223144JPEG2025538495000081.jpg243159JPEG2025538495000082.jpg253159TIFF2025538495000083.tif242156TIFF2025538495000084.tif236159TIFF2025538495000085.tif231159JPEG2025538495000086.jpg236159JPEG2025538495000087.jpg250159TIFF2025538495000088.tif242158TIFF2025538495000089.tif238159TIFF2025538495000090.tif236159JPEG2025538495000091.jpg246159JPEG2025538495000092.jpg251159TIFF2025538495000093.tif234159JPEG2025538495000094.jpg236159TIFF2025538495000095.tif242158TIFF2025538495000096.tif236159JPEG2025538495000097.jpg242159TIFF2025538495000098.tif242158TIFF2025538495000099.tif234159TIFF2025538495000100.tif230159TIFF2025538495000101.tif242157TIFF2025538495000102.tif234159JPEG2025538495000103.jpg252159JPEG2025538495000104.jpg236159JPEG2025538495000105.jpg250159JPEG2025538495000106.jpg237159JPEG2025538495000107.jpg251159JPEG2025538495000108.jpg235159JPEG2025538495000109.jpg251159JPEG2025538495000110.jpg244159TIFF2025538495000111.tif238159TIFF2025538495000112.tif236159JPEG2025538495000113.jpg249159JPEG2025538495000114.jpg249159.

[0075] In one embodiment, the ionizable lipid of the present disclosure is tridecyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0099), 8-methylnonyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0101), 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)tridecyl propanoate (VC-LC-0163), tetradecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0168), octadecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0169), 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl )thio)hexadecyl propanoate (VC-LC-0170), tridecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0178), tetradecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0183), 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)propanoate tridecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0194), 8-methylnonyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0196), dodecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0198),Tetradecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0199), octadecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0200), 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido) )-4-oxobutyl)thio) hexadecyl propanoate (VC-LC-0201), 6-methylheptyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0202), octyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0207), tridecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0209), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0211), dodecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0213), tetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0214), octadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0215), hexadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0216),Tridecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((3-(pyrrolidin-1-yl)propyl)amino)butyl)thio)propanoate (VC-LC-0241), Tridecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0256), 8-methyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate Nyl (VC-LC-0258), 8-methylnonyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0261), octadecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0262), 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)hexadecyl propanoate (VC-LC-0263), 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)hexyl propanoate (VC-LC-0268), 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)octyl propanoate (VC-LC-0269), 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3 -oleamido-4-oxobutyl)thio)propanoic acid tridecyl ester (VC-LC-0289), 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butanoic acid heptadecan-9-yl ester (VC-LC-0294), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid heptadecan-9-yl ester (VC-LC-0296),4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid heptadecan-9-yl ester (VC-LC-0297), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid heptadecan-9-yl ester (VC-LC-0298), 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-( Heptadecan-9-yl ((3-oxo-3-(tetradecyloxy)propyl)thio)methyl)butanoate (VC-LC-0299), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0300), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate -yl (VC-LC-0301), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate heptadecan-9-yl (VC-LC-0302), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate heptadecan-9-yl (VC-LC-0304), 4-((3-(1H-imidazol-1-yl)propyl Heptadecan-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0306), Heptadecan-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0307), 2-decyltetradecyl 3-((4-((3-morpholinopropyl)amino)-3-oleamido-4-oxobutyl)thio)propanoate (VC-LC-0353),Octadec-9-en-1-yl 3-((4-((3-morpholinopropyl)amino)-3-oleamido-4-oxobutyl)thio)propanoate (VC-LC-0355), 2-decyltetradecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-oleamido-4-oxobutyl)thio)propanoate (VC-LC-0356), heptadecyl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoate Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-(dodecyloxy)-3-oxopropyl)thio)butanoate (VC-LC-0366), Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butanoate (VC-LC-0367), 2-((3-(1H-imidazol-1-yl) Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)butanoate (VC-LC-0369), Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)butanoate (VC-LC-0370), 2-decyl 3-((4-((3-morpholinopropyl)amino)-4-oxo-3-stearamidobutyl)thio)propanoate tetradecyl (VC-LC-0389), 2-hexyldecyl 6-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-6-oxohexanoate (VC-LC-0418), 2-decyltetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0428),Dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0429), octadec-9-en-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0430), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0431), 4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-2-((, Heptadecan-9-yl 1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0439), 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0440), 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoate (VC-LC-0441), 4-((3-(dec-2-yn-1-yloxy)-3 Heptadecan-9-yl (Z)-2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)butanoate (VC-LC-0442), Heptadecan-9-yl (Z)-2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)butanoate (VC-LC-0443), Heptadecan-9-yl 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butanoate (VC-LC- -0444), tridecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0473), 2-hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0474), 8-methylnonyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC -0475), dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0477), tetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0478), hexadecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0480),2-Decyltetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0487), Dec-2-yn-1-yl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0488), Octadec-9-ene 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)tridecyl propanoate (VC-LC-0490), 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)2-hexyldecyl propanoate (VC-LC-0491), 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)- 8-Methylnonyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0492), 2-Decyltetradecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0504), Dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0505), 3-((4-(((1-methylpiperidin-4- tridecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0507), 2-hexyldecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0508), 8-methylnonyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0509),2-Ethylhexyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0510), 6-methylheptyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0515), 6-methylheptyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0516), 2-Decyltetradecyl 4-((1-methylpiperidin-4-yl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butanoate (VC-LC-0524), 2-Hexyldecyl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutanoate (VC-LC-0525), 2-(((3-( 2-Hexyldecyl (Z)-4-((1-methylpiperidin-4-yl)amino)-2-(((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0531), 2-Hexyldecyl (Z)-4-((1-methylpiperidin-4-yl)amino)-2-(((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0539), 4-((3-morpholinopropyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyl) 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0540), 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0541), 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0542),2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0543), 2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0544), 4-((3-morpholinopropyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)amino) Heptadecan-9-yl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0546), Heptadecan-9-yl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0547), Heptadecan-9-yl 2-(((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0548) , 2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoic acid heptadecan-9-yl ester (VC-LC-0549), 2-(((3-(hexyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoic acid heptadecan-9-yl ester (VC-LC-0550), 4-((3-morpholinopropyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl 2-(((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0551), 2-(((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0553), 2-(((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0554),Tridecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0556), 2-hexyldecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0557), 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate 8 -methylnonyl (VC-LC-0558), 2-ethylhexyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0559), 2-hexyldecyl 4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0606), 4-((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)-2-((3- 2-Hexyldecyl 4-((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0607), 2-Hexyldecyl 4-((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0608), 2-Hexyldecyl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0609), 2-((3-morpholinopropyl)carbamoyl)-4- 2-hexyldecyl ((3-oxo-3-(tetradecyloxy)propyl)thio)butanoate (VC-LC-0610), 2-hexyldecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0639), 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0729),8-Methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0730), 2-ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0731), -((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)dodecyl propanoate (VC-LC-0732), 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)tetradecyl propanoate (VC-LC-0733), 3-((4-((2-(dimethylamino), Octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0734), hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0735), 6-methylheptyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0736), Docosyl 2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0737), hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0738), octyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0739), 2-Decyltetradecyl 2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0741), Dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0742), Octadec-9-ene 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate -1-yl (VC-LC-0743), 8-(2-((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)ethyl)-3-ethyl-15-((3-(octyloxy)-3-oxopropyl)thio)-7,10-dioxo-18-thia-3,6,9-triaza-21-heneicosylate octyl (VC-LC-0757), 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate tridecyl (VC-LC-0796),2-Hexyldecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0797), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0798), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0799), 2-Ethylhexyl propanoate (VC-LC-0799), dodecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0800), tetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0801), 2-ethylhexyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0802), Octadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0802), hexadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0803), 6-methylheptyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0804), 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoic acid docosyl ester (VC-LC-0805), 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoic acid hexyl ester (VC-LC-0806), 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoic acid octyl ester (VC-LC-0807),2-Decyltetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0809), dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0810), 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl )thio)propanoic acid octadec-9-en-1-yl ester (VC-LC-0811), 3-((4-(4-(2-hydroxyethyl)piperazin-1-yl)-3-(2-octyldodecanamido)-4-oxobutyl)thio)tridecyl ester (VC-LC-0828), 3-((3-(2-octyldodecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoic acid 8-methylnonyl ester (VC-LC-0848), 3-((4-((2-(dimethylamino)ethyl)amino )-3-(2-octyldodecanamido)-4-oxobutyl)thio)tridecyl propanoate (VC-LC-0864), 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0865), 8-methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0866), 3-((4-((2-( 2-Ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0867), dodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0868), tetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0869),Octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0870), hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0871), 6-methyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate Butyl (VC-LC-0872), docosyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0873), hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0876), 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio) Octyl propanoate (VC-LC-0877), dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0880), octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0881), 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)propanoate 6-methylheptyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0890), octyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0895), 2-hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0919),2-Ethylhexyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0921), dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0922), tridecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-09 36), 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0937), dodecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0940), 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate 6 -methylheptyl (VC-LC-0944), docosyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0945), octyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0949), 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxo (Z)-octadec-9-en-1-yl 6-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-6-oxohexanoate (VC-LC-0973), 2-octyldodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1065),7-(2-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)ethyl)-2-methyl-14-((3-(octyloxy)-3-oxopropyl)thio)-6,9-dioxo-17-thia-2,5,8-triaza-20-eicosanoic acid octyl ester (VC-LC-1068), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-1-oxo-4-((3-oxopropyl)amino)-2-methyl-14-((3-(octyloxy)-3-oxopropyl)thio)-6,9-dioxo-17-thia-2,5,8-triaza-20-eicosanoic acid octyl ester (VC-LC-1068), dioctyl 3-(tridecyloxy)propyl)thio)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl)dipropionate (VC-LC-1082), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino, )-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1083), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate dioctyl (VC-LC-1084), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate dioctyl (VC-LC-1084) )propyl)amino)-4-((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1085), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(octadecyloxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate dioctyl dipropionate (VC-LC-1088), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(hexadecyloxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl)) dioctyl dipropionate (VC-LC-1089), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1090), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1091), 3,Dioctyl 3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1094), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1094), Dioctyl 3,3'-((8-oxo-8-((1-oxo-4-((3-oxo-3-(tridecyloxy)propyl)thio)-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)octane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1096) ), 3,3'-((8-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1097), 3,3'-((8-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino) )amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1098), 3,3'-((8-((4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1105), 3,3'-((8-((4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))(Z)-dipropionic acid dioctyl ester (VC-LC-1111), 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoic acid (9Z,12Z)-octadeca-9,1 2-dien-1-yl (VC-LC-1143), (2E,6E)-3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoic acid 3,7,11-trimethyldodeca-2,6,10-trien-1-yl (VC-LC-1192), bis((E)-2-(((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)methyl)succinic acid 3,7-dimethylocta-2,6-dien-1-yl) VC-LC-1194), tetradecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1202), dodecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1203), 2-(((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecane) Bis(3,7-dimethyloctyl) 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)methyl)succinate (VC-LC-1218), 2-hexyldecyl 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1224), (9Z,12Z)-octadeca-9 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate,12-dien-1-yl (VC-LC-1230), 2-((2-(dimethylamino)ethyl)carbamoyl)-4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)butanoate 2-hexyldecyl (VC-LC-1254), 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyl)oxy)carbonyl)-4-oxobutyl)thio)methyl 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyl)oxy)carbonyl)-4-oxobutyl)thio)methyl)succinate bis((E)-3,7-dimethylocta-2,6-dien-1-yl) (VC-LC-1282), 2-(((4-((2-butyloctyl)oxy)-3- Bis((E)-3,7-dimethylocta-2,6-dien-1-yl) ((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1291), bis(3,7-dimethyloctyl) 2-(((4-((2-decyltetradecyl)oxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC -LC-1295), 2-(((4-((2-decyltetradecyl)oxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate bis((E)-3,7-dimethylocta-2,6-dien-1-yl) (VC-LC-1296), or a pharmaceutically acceptable salt thereof, or any one stereoisomer thereof.

[0076] In another embodiment, the ionizable lipid of formula (I) is VC-LC-0099, VC-LC-0101, VC-LC-0163, VC-LC-0168, VC-LC-0169, VC-LC-0170, VC-LC-0178, VC-LC-0183, VC-LC-0194, VC-LC-0196, VC-LC-0198, VC-LC-0199, VC-LC-0200, VC-LC-0201, VC-LC-0202, VC-LC-0207, VC-LC-0209, VC-LC-0211, VC-LC-0213, VC-LC-0214, VC-LC-0215, VC-LC-0216, VC-LC-0241, VC-LC-0256, VC-LC-0258, VC-LC-0261, VC-LC-0262, VC-LC-0263, VC-LC-0268, VC-LC-0269, VC-LC-0289, VC-LC-0294, VC-LC-0296, VC-LC-0297, VC-LC-0298, VC-LC-0299, VC-LC-0300, VC-LC-0301, VC-LC-0302, VC-LC-0304, VC-LC-0306, VC-LC-0307, VC-LC-0353, VC-LC-0355, VC-LC-0356, VC-LC-0362, VC-LC-0366, VC-LC-0367, VC-LC-0369, VC-LC-0370, VC-LC-0389, VC-LC-0418, VC-LC-0428, VC-LC-0429, VC-LC-0430, VC-LC-0431, VC-LC-0439, VC-LC-0440, VC-LC-0441, VC-LC-0442, VC-LC-0443, VC-LC-0444, VC-LC-0473, VC-LC-0474, VC-LC-0475, VC-LC-0477, VC-LC-0478, VC-LC-0480, VC-LC-0487, VC-LC-0488, VC-LC-0489, VC-LC-0490, VC-LC-0491, VC-LC-0492, VC-LC-0504, VC-LC-0505, VC-LC-0507, VC-LC-0508, VC-LC-0509, VC-LC-0510, VC-LC-0515, VC-LC-0521, VC-LC-0524, VC-LC-0525, VC-LC-0531, VC-LC-0539, VC-LC-0540, as described herein.VC-LC-0541, VC-LC-0542, VC-LC-0543, VC-LC-0544, VC-LC-0546, VC-LC-0547, VC-LC-0548, VC-LC-0549, VC-LC-0550, VC-LC-0551, VC-LC-0553, VC-L C-0554, VC-LC-0556, VC-LC-0557, VC-LC-0558, VC-LC-0559, VC-LC-0606, VC-LC-0607, VC-LC-0608, VC-LC-0609, VC-LC-0610, VC-LC-0639, VC-LC-07 29, VC-LC-0730, VC-LC-0731, VC-LC-0732, VC-LC-0733, VC-LC-0734, VC-LC-0735, VC-LC-0736, VC-LC-0737, VC-LC-0738, VC-LC-0739, VC-LC-0741, V C-LC-0742、VC-LC-0743、VC-LC-0757、VC-LC-0796、VC-LC-0797、VC-LC-07 98、VC-LC-0799、VC-LC-0800、VC-LC-0801、VC-LC-0802、VC-LC-0803、VC-LC -0804、VC-LC-0805、VC-LC-0806、VC-LC-0807、VC-LC-0809、VC-LC-0810、VC-LC-0811、VC-LC-0828、VC-LC-0848、VC-LC-0864、VC-LC-0865、VC-LC-086 6, VC-LC-0867, VC-LC-0868, VC-LC-0869, VC-LC-0870, VC-LC-0871, VC-LC-0872, VC-LC-0873, VC-LC-0876, VC-LC-0877, VC-LC-0880, VC-LC-0881, VC -LC-0890、VC-LC-0895、VC-LC-0919、VC-LC-0921、VC-LC-0922、VC-LC-093 6、VC-LC-0937、VC-LC-0940、VC-LC-0944、VC-LC-0945、VC-LC-0949、VC-LC- 0953, VC-LC-0973, VC-LC-1065, VC-LC-1068, VC-LC-1082, VC-LC-1083, VC-LC-1084, VC-LC-1085, VC-LC-1088, VC-LC-1089, VC-LC-1090, VC-LC-1091VC-LC-1094, VC-LC-1095, VC-LC-1096, VC-LC-1097, VC-LC-1098, VC-LC-1105, VC-LC-1111, VC-LC-1143, VC-LC-1192, VC-LC-1194, VC-LC-1202, VC-LC-1203, VC-LC-1218, VC-LC-1224, VC-LC-1230, VC-LC-1254, VC-LC-1281, VC-LC-1282, VC-LC-1291, VC-LC-1295, VC-LC-1296, or a pharmaceutically acceptable salt thereof, or a compound selected from the group consisting of any one stereoisomer thereof.

[0077] In certain embodiments, the ionizable lipid of formula (I) is VC-LC-0263, VC-LC-0418, VC-LC-0431, VC-LC-0474, VC-LC-0478, VC-LC-0489, VC-LC-0508, VC-LC-554, VC-LC-559, VC-LC-0639, VC-LC-0729, VC-LC-0730, VC-LC-0731, VC-LC-0732, VC-LC-0733, VC-LC-0734, VC-LC-0735, VC-LC-0736, VC-LC-0737, VC-LC-0738, VC-LC-0739, VC-LC-0741, VC-LC-0742, VC-LC-0743, VC-LC-0757, VC-LC-0796, VC-LC-0797, VC-LC-0798, VC-LC-0799, VC-LC-0800, VC-LC-0801, VC-LC-0802, VC-LC-0803, VC-LC-0804, VC-LC-0805, VC-LC-0806, VC-LC-0807, VC-LC-0809, VC-LC-0810, VC-LC-0811, VC-LC-0828, VC-LC-0848, VC-LC-0864, VC-LC-0865, VC-LC-0866, VC-LC-0867, VC-LC-0868, VC-LC-0869, VC-LC-0870, VC-LC-0871, VC-LC-0872, VC-LC-0873, VC-LC-0876, VC-LC-0877, VC-LC-0880, VC-LC-0881, VC-LC-0890, VC-LC-0895, VC-LC-0919, VC-LC-0921, VC-LC-0922, VC-LC-0936, VC-LC-0937, VC-LC-0940, VC-LC-0944, VC-LC-0945, VC-LC-0949, VC-LC-0953, VC-LC-0973, VC-LC-1065, VC-LC-1068, VC-LC-1082, VC-LC-1083, VC-LC-1084, VC-LC-1085, VC-LC-1088, VC-LC-1089, VC-LC-1090, VC-LC-1091, VC-LC-1094, VC-LC-1095, VC-LC-1096, VC-LC-1097, VC-LC-1098, VC-LC-1105, VC-LC-1111, VC-LC-1143, VC-LC-1192, as described herein.VC-LC-1194, VC-LC-1202, VC-LC-1203, VC-LC-1218, VC-LC-1224, VC-LC-1230, VC-LC-1254, VC-LC-1281, VC-LC-1282, VC-LC-1291, VC-LC-1295, VC-LC-1296, or a pharmaceutically acceptable salt thereof, or a compound selected from the group consisting of any one stereoisomer thereof.

[0078] Preparation Process Processes for the preparation of ionizable lipids of formula (I) are also part of the present invention.

[0079] All reactants and solvents required for the preparation of the compounds of the present disclosure are commercially available.

[0080] By way of example, ionizable lipids of formula (I) can be prepared according to any of the following synthetic schemes: One skilled in the art will know which reactants are necessary to obtain any particular ionizable lipid according to the present disclosure according to the synthetic methods shown in the following schemes or methods similar thereto.

[0081] Ionizable lipids of formula (II) where B is an amide moiety and t=0, q=2, such as compounds of formula (IIa), can be prepared by the synthetic method shown in Scheme 1 below or by analogous methods. [ka]

[0082] Acids (R 1 Examples of amines (AM-NH2), amines (AM-NH2), and acrylates (CH2=CH-(CO)-O-R2) are listed below in Tables 2, 3, and 4, respectively. [Table 2] [Table 3] TIFF2025538495000118.tif48159 [Table 4]

[0083] Ionizable lipids of formula (II) where B is an amide moiety and t=q=1, such as compounds of formula (IIb) or analogs thereof, can be prepared by the synthetic method shown in Scheme 2 below. [ka]

[0084] Ionizable lipids of formula (I) where B is an ester moiety, such as, for example, a compound of formula (IIc) or an analogous compound thereof or a compound of formula (IId) or an analogous compound thereof, can be prepared by the synthetic methods shown in Schemes 3 or 4 below or by analogous methods.

[0085] Lipid analogs with amide-ester substitutions, i.e., compounds of formula (IIc) where B=C, t=0, q=2 and compounds of formula (IId) where B=C, t=q=1, can be prepared according to the following synthetic schemes 3 and 4: [ka] [ka]

[0086] Depending on whether the reaction is carried out starting from 2-oxotetrahydrothiophene-3-carboxylic acid (compound A) or 5-oxotetrahydrothiophene-3-carboxylic acid (compound B), two different families of compounds (i.e., compounds of formula (IIc) or (IId)), respectively, can be obtained. 2-Oxotetrahydrothiophene-3-carboxylic acid and 5-oxotetrahydrothiophene-3-carboxylic acid can be prepared as described in the literature (Garbiras, BJ; Marburg, S. "Preparation of Carboxythiolactones and Their Active Derivatives", Synthesis, 1999, vol. 2, pp. 270-274).

[0087] Possible alcohols R used for the preparation of compounds of formula (IIc) and formula (IId) according to schemes 3 and 4 4 The -OH are listed below in Table 5. The amines AM-NH2 and acrylates CH2=CH(CO)OR2 are those listed above in Tables 3 and 4. [Table 5]

[0088] Ionizable lipids of formula (I) where B is an amide moiety, such as compounds of formula (IIe) (A=B=amide, t=2, q=1) or analogous compounds thereof, can be prepared by the synthetic method shown in Scheme 5 below or analogous methods. [ka]

[0089] Ionizable lipids of formula (I) where B is an amide moiety, such as, for example, a compound of formula (IIf) (A=B=amide, t=0, q=3) or similar compounds, can be prepared by the synthetic method shown in Scheme 6 below or similar methods. [ka]

[0090] Compounds C and D are commercially available.

[0091] Ionizable lipids of formula (I) or similar compounds, where B is an amide moiety (A=B=amide), t=0 and q=1, can be prepared as disclosed in French Patent Application Publication No. 3104943A1.

[0092] For example, Z is R1, X is [ka] Ionizable lipids of formula (I), such as compounds of formula (III) where: and compounds of formula (IIIa) where: A=B=amide, t=0, q=3, or similar compounds thereof, can be prepared by the synthetic method shown in Scheme 7 below or similar methods. [ka]

[0093] For example, X is -D-R2, and Z is [ka] Ionizable lipids of formula (I), such as compounds of formula (IV) where: and compounds of formula (IVa) where: A=B=—NH—C(O)—, t=0, q=2, or similar compounds thereof, can be prepared by the synthetic method shown in Scheme 8 below or similar methods. [ka]

[0094] lipid nanoparticles As mentioned above, the ionizable lipids of the present disclosure can form LNPs in solution.

[0095] Thus, the present invention also relates to LNPs comprising ionizable lipids of formula (I) as defined herein (particularly lipids of formula (II), (III) or (IV)), including ionizable lipids not claimed above or below.

[0096] All embodiments shown for the ionizable lipids of formula (I) also apply to LNPs.

[0097] Typically, LNPs have a core-shell structure comprising an inner core and an outer shell. LNP formulations containing several lipid components of different nature, such as ionizable lipids, sterols, PEG lipids, and non-cationic lipids such as phospholipids (also referred to as "helper lipids"), comprise several phases: a hydrophobic core region formed by ionizable lipids and cholesterol, and a surrounding shell formed by helper lipids, cholesterol, and PEG lipids covering the surface. More specifically, the ionizable lipid of formula (I) forms part of the inner shell of the LNP, and optionally, a pharmaceutically active agent is encapsulated or loaded into the inner core.

[0098] In one embodiment, in any combination with one or more features of the various embodiments described above, the LNP further comprises at least one lipid selected from the group consisting of a non-cationic lipid, a sterol, or a steroid precursor, and a PEG-modified lipid.

[0099] In another embodiment, optionally in combination with one or more features of the various embodiments described above, the LNP comprises an ionizable lipid of the present disclosure and a non-cationic lipid, examples of which include, but are not limited to, 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SO ... Palmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 Examples of suitable non-cationic lipids include 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerin) sodium salt (DOPG), sphingomyelin, and mixtures thereof. In one embodiment, the non-cationic lipid is DSPC.In certain embodiments, the non-cationic lipid is DOPE. In another particular embodiment, the lipid-containing particle comprises DSPC and DOPE.

[0100] In another embodiment, in any combination with one or more features of the various embodiments described above, the LNP further comprises a sterol or a sterol precursor.

[0101] In another embodiment, in any combination with one or more features of the various embodiments described above, the LNP comprises an ionizable lipid of the present disclosure and a sterol or sterol precursor.

[0102] Examples of sterols include, but are not limited to, cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof. In certain embodiments, optionally in combination with one or more features of the various embodiments described above, the sterol is cholesterol. Examples of sterol precursors include, but are not limited to, triterpenes, triterpenoids, or steroid precursors of this type. Non-limiting examples of triterpenes, triterpenoids, and other steroid precursors include squalene, achilleol, polypodatetrane, lanostane, cucurbitacin, hopane, oleanane, chamaecydin, lupine, and mixtures thereof.

[0103] In another embodiment, in any combination with one or more features of the various embodiments described above, the LNP comprises an ionizable lipid of the present disclosure, a non-cationic lipid as defined above, and a sterol or sterol precursor as defined above.

[0104] In another embodiment, optionally in combination with one or more features of the various embodiments described above, the LNP further comprises a PEG-modified lipid. The term "PEG-modified lipid" refers to a lipid containing a polyethylene moiety. Examples of PEG-modified lipids include, but are not limited to, PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified phosphatidylcholine, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerin, and mixtures thereof. In some embodiments, the PEG-modified lipid is PEG-DMG, PEG-c-DOMG (also referred to as PEG-DOMG), PEG-DSG, PEG-DPG, or a combination thereof.

[0105] In another embodiment, in any combination with one or more features of the various embodiments described above, the LNP further comprises a conjugated lipid.

[0106] Examples of conjugated lipids include, but are not limited to, polysarcosine (pSar) lipids and derivatives such as N-tetradecyl-pSar25, N-hexadecyl-pSar25, N-octadecyl-pSar25, N-dodecyl-pSar25, DMG-pSar25, 18:1 PE(DOPE)pSar25, N-TETAMINE-pSar25, N-TETAMINE-pSar35, N-TETAMINE-pSar45, and N-TETAMINE-pSar45-maleimide.

[0107] In another embodiment, in any combination with one or more features of the various embodiments described above, the LNP comprises an ionizable lipid of the present disclosure, a non-cationic lipid as defined above, and a PEG-modified lipid or conjugated lipid as defined above.

[0108] In one embodiment, in any combination with one or more features of the various embodiments described above, the LNP comprises a lipid component that includes or consists of an ionizable lipid, a non-cationic lipid, a sterol, and a PEG-modified lipid as disclosed herein.

[0109] In certain embodiments, in any combination with one or more features of the various embodiments described above, the LNP comprises a lipid component comprising an ionizable lipid disclosed herein and at least one of distearoylphosphatidylcholine (DSPC), cholesterol, and DMG-PEG 2000. In more particular embodiments, the lipid component comprises or consists of an ionizable lipid disclosed herein, DSPC, cholesterol, and DMG-PEG 2000.

[0110] In another embodiment, optionally in combination with one or more features of the various embodiments described above, the LNP comprises 25-60 mol% ionizable lipid, 0.1-10 mol% PEG-modified lipid or alternatively complex lipid (particularly PEG-modified lipid), 10-45 mol% non-cationic lipid, and 10-40 mol% sterol. Specifically, the LNP comprises 32-50 mol% ionizable lipid, 1-8 mol% PEG-modified lipid or complex lipid (particularly PEG-modified lipid), 12.5-42 mol% non-cationic lipid, and 15-38.5 mol% sterol. More specifically, the LNP comprises 35-47 mol% ionizable lipid, 1-4 mol% PEG-modified lipid, 30-38 mol% non-cationic lipid, and 15-25 mol% sterol.

[0111] In another embodiment of the LNP, in any combination with one or more features of the various embodiments described above, the ionizable lipid is in an amount of 25-64 mol %, the PEG-modified lipid is in an amount of 0.1-1.5 mol %, and the sterol is in an amount of 35-74 mol %.

[0112] As used herein, "mol %" refers to the mole % of a component relative to the total moles of all lipid components in the LNP (i.e., the total moles of ionizable lipids, PEG-modified lipids, non-cationic lipids, and sterols).

[0113] In another embodiment, in any combination with one or more features of the various embodiments described above, the molar ratio of ionizable lipid:non-cationic lipid ranges from 6:1 to 1:2 or from 2:1 to 1:1.

[0114] In another embodiment, in any combination with one or more features of the various embodiments described above, the molar ratio of ionizable lipid:sterol ranges from 5:1 to 1:2 or from 2:1 to 1:1.

[0115] In another embodiment, in any combination with one or more features of the various embodiments described above, the molar ratio of ionizable lipid:PEG-modified lipid ranges from 120:1 to 2:1 or from 100:1 to 10:1.

[0116] activator As noted above, the LNPs of the present invention may include one or more pharmaceutically active agents.

[0117] As used herein, the term "pharmaceutically active agent" refers to an agent that has pharmacological activity and is used to cure, alleviate, treat or prevent disease in a subject, particularly a human.

[0118] Pharmaceutically active agents for purposes of the present invention include low molecular weight drugs, polynucleotides, peptides, antibodies, proteins and combinations thereof.

[0119] The term "polynucleotide" as used herein refers to natural or artificial deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). Polynucleotides include pseudouridine, N1-methylpseudouridine (also known as 1-methylpseudouridine or m1Ψ), N6-methyladenosine (also known as m6A), 2-thiouridine (also known as s2U), 4'-thiouridine, 5-methylcytosine (also known as 5mC), 2-thio-1-methyl-1-deazapseudouridine, 2-thio-1-methylpseudouridine, 2-thio-5-azauridine, 2-thiodihydropseudouridine, The compound may comprise at least one chemical modification selected from the group consisting of 2-thiodihydrouridine, 2-thiopseudouridine, 4-methoxy-2-thiopseudouridine, 4-methoxypseudouridine, 4-thio-1-methylpseudouridine, 4-thiopseudouridine, 5-azauridine, dihydropseudouridine, 5-methyluridine (also referred to as m5U), 5-methoxyuridine, 2'-O-methyluridine, and combinations thereof. In particular, the chemical modification is N1-methylpseudouridine, 5-methoxyuridine, or a combination thereof, and in particular, the chemical modification is N1-methylpseudouridine.

[0120] The polynucleotide is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 99% partially modified with N1-methylpseudouridine or is fully modified with N1-methylpseudouridine, 5-methoxyuridine, or a combination thereof.

[0121] In certain embodiments, optionally in combination with any of the embodiments provided above, the active agent is selected from the group consisting of a polynucleotide, a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide, and an expression vector comprising a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide.

[0122] In certain embodiments, optionally in combination with any of the embodiments provided above, the polynucleotide is ribonucleic acid (RNA).

[0123] In particular, the RNA is selected from the group consisting of short interfering RNA (siRNA), self-replicating RNA (srRNA), circular RNA (circRNA), self-amplifying RNA (saRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), small interfering RNA (siRNA), small RNA (sRNA), Dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof.

[0124] In certain embodiments, optionally in combination with any of the embodiments provided above, the RNA is mRNA.

[0125] A person skilled in the art knows how to make a polynucleotide, a DNA construct or an expression vector by routine methods well known in the art without the exercise of inventive skill, such as chemical synthesis or molecular biology techniques.

[0126] In certain embodiments, optionally in combination with any of the embodiments provided above, the ionizable lipid:RNA ratio (N / P; where N represents the moles of amine present in the ionizable lipid and P represents the moles of phosphate present in the polynucleotide backbone) in the LNP ranges from 20:1 to 2:1, particularly 10:1 to 3:1.

[0127] In certain embodiments, optionally in combination with any of the embodiments provided above, the polynucleotide is an isolated, man-made polynucleotide.

[0128] In one embodiment, in any combination with one or more features of the various embodiments described above, the LNP comprises an ionizable lipid, a non-cationic lipid, a sterol, a PEG-modified lipid, and a polynucleotide as defined herein.

[0129] The LNPs containing one or more polynucleotides may be prepared by standard methods, such as microfluidic mixing as disclosed in Hassett, KJ et al., "Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines," 2019, Mol. Ther. Nucleic Acid, vol. 15, pp. 1-11, or manual / bulk mixing as disclosed in Wang X., Liu S., Sun Y. et al., "Preparation of selective organ-targeting (SORT) lipid nanoparticles (LNPs) using multiple technical methods for tissue-specific mRNA delivery," 2022, Nat. Protoc., doi:10.1038 / s41596-022-00755-x. Both methods are known in the art and the skilled person will know how to proceed in each specific case.

[0130] Typically, the process for preparing LNPs includes: (i) preparing a first alcohol mixture containing the ionizable lipids of the present disclosure and, optionally, at least one lipid selected from the group consisting of non-cationic lipids, sterols, and PEG-modified lipids in a suitable alcohol, such as ethanol; (ii) preparing a second aqueous composition containing a polynucleotide and an acidified buffer; and (iii) mixing (i) with (ii) in a microfluidic mixer. The microfluidic mixer allows thorough and rapid mixing of the lipid phase and the polynucleotide phase in a microscale device. Depending on the process parameters (especially the total flow rate), those skilled in the art will be able to adjust the size of the LNPs.

[0131] In certain embodiments, optionally in combination with any of the embodiments provided above, the polynucleotide encodes a polypeptide (particularly where the polypeptide is an antigen), more particularly, the antigen is selected from the group consisting of a viral protein, a bacterial protein, and a tumor-associated antigen.

[0132] In certain embodiments, optionally in combination with any of the embodiments provided above, the polypeptide is an antibody or fragment thereof. In more particular embodiments, the antibody or fragment thereof is a therapeutic antibody or fragment thereof.

[0133] In another embodiment, optionally in combination with any of the embodiments provided above, the antigen is a SARS-CoV-2 antigen, in particular a SARS-CoV-2 spike antigen.

[0134] The preparation methods of the above-mentioned lipids, LNPs and pharmaceutical compositions are described herein and / or known in the art.Those skilled in the art will know which LNPs to use for encapsulating each pharmaceutically active agent according to the intended use of this composition.In particular, the method for synthesizing the composition formed by this LNP that encapsulates RNA is well known to those skilled in the art and is well established in molecular biology protocols.Specific conditions are shown in Examples.

[0135] As indicated above, another aspect of the present invention relates to pharmaceutical compositions comprising LNPs as defined herein and a pharmaceutically acceptable excipient or carrier.

[0136] The phrase "pharmaceutically acceptable excipient or carrier" refers to a pharmaceutically acceptable material, composition, or vehicle. Each component must be pharmaceutically acceptable in the sense of being compatible with the other components of the pharmaceutical composition. It must also be suitable for use in contact with the tissues or organs of humans and non-human animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0137] Examples of suitable pharmaceutically acceptable excipients are solvents, dispersion media, diluents or other liquid vehicles, dispersants, suspending aids, surfactants, isotonicity agents, thickeners, emulsifiers, preservatives, solid binders, lubricants, etc. Except insofar as any conventional excipient medium is incompatible with a substance or its derivatives, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component of the pharmaceutical composition, its use is contemplated within the scope of the present invention.

[0138] The relative amounts of the pharmaceutically active ingredient, pharmaceutically acceptable excipient and / or any additional ingredients in a pharmaceutical composition of the present invention will vary depending on the identity, size and / or condition of the subject being treated, and also depending on the route by which the composition is administered.

[0139] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surface active agents and / or emulsifying agents, disintegrating agents, binders, preservatives, buffers, lubricants and / or oils. Excipients such as coloring agents, coating agents, sweetening agents and flavoring agents may also be present in the composition, according to the judgment of the formulator.

[0140] In one embodiment, optionally in combination with any of the embodiments provided above, the pharmaceutical compositions disclosed herein are administered by oral, intranasal, intravenous, intraperitoneal, intramuscular, intradermal, subcutaneous, topical, or intraarticular administration.

[0141] The pharmaceutical compositions of the present disclosure may be prepared by methodologies well known in the pharmaceutical industry. For example, pharmaceutical compositions intended for administration by injection can be prepared by combining the lipid nanoparticles of the present invention with sterile distilled water or other carriers to form a solution. Some excipients or carriers can be added to facilitate the formation of a homogeneous solution or suspension. As described above, the LNPs or pharmaceutical compositions of the present disclosure containing pharmaceutically active agents can be used for therapeutic purposes. In particular, they can be used as non-viral vectors commonly used for biomedical applications such as vaccines or gene therapy, which are effective in transfecting genetic material into eukaryotic cells.

[0142] Accordingly, one aspect of the present invention relates to a LNP or pharmaceutical composition as defined herein for use in a method of treating a disease or disorder in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a nanoparticle composition or pharmaceutical composition as defined herein.

[0143] Another aspect of the invention relates to an LNP or pharmaceutical composition as defined herein for use in a method for inducing an immune response in a subject, for use in a method for therapeutic immunization of a subject, for use as a vaccine, or for use in gene therapy, particularly wherein the subject is a human.

[0144] In one embodiment, optionally in combination with any of the embodiments provided above, the disease or disorder is selected from the group consisting of an infectious disease, a cancer, a proliferative disease, a genetic disease, an autoimmune disease, diabetes, a neurodegenerative disease, a cardiovascular disease, a renal vascular disease, and a metabolic disease.

[0145] When the pharmaceutically active agent is a polynucleotide, the LNPs or pharmaceutical compositions described herein can be used in vaccine therapy, in enhancing the effectiveness of conventional vaccines, and / or as novel vaccine forms for use against infectious pathogens such as viruses, bacteria, fungi, protozoa, prions, and helminths (parasites), or for use in treating diseases such as cancer and proliferative disorders.

[0146] In one embodiment, optionally in combination with any of the embodiments provided above, the pharmaceutical composition is a vaccine.In a more particular embodiment, the pharmaceutical composition is a vaccine and further comprises an adjuvant.Those skilled in the art will know based on their general and general knowledge which excipients, carriers and adjuvants to include in the vaccine according to the intended use.

[0147] Throughout the specification and claims, the term "comprises" and variations of that term are not intended to exclude other technical features, additives, ingredients or steps. Furthermore, the term "including" encompasses the case of "consisting of."

[0148] The following examples and figures are given by way of illustration and are not intended to limit the invention. Furthermore, the present invention includes all possible combinations of particular and preferred embodiments described herein. [Example]

[0149] Reagents were purchased from Sigma-Aldrich, TCI Chemicals, Fluorochem, or VWR. All possible combinations shown above were prepared following a three-step reaction protocol.

[0150] Example 1: Synthesis of lipid VC-LC-0163 [ka]

[0151] Process 1 DL-Homocysteine ​​hydrochloride (776 mg, 5.0 mmol) was dissolved in 13 mL of anhydrous methylene chloride at room temperature. Triethylamine (588 μL, 4.3 mmol) was then added, followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (870 mg, 4.5 mmol), dimethylaminopyridine (104 mg, 0.85 mmol), and 2-hexyldecanoic acid (1.30 g, 4.3 mmol). The reaction mixture was stirred overnight at room temperature and then checked for completion by TLC chromatography. The resulting product was purified by flash chromatography using a hexane / ethyl acetate gradient (100 / 0 to 0 / 100). Fractions containing the product were combined and evaporated under reduced pressure to give a yellowish oil (1.39 g, 89%).

[0152] Process 2 The product from step 1 (250 mg, 0.7 mmol) was dissolved in anhydrous tetrahydrofuran (1.5 mL) and added to a solution of 3-morpholinopropylamine (1.03 mL, 7.03 mmol) in anhydrous tetrahydrofuran (1.0 mL) under a nitrogen atmosphere. The solution was then stirred at room temperature for 30 minutes, after which the solvent was evaporated under reduced pressure. The dried mixture was redissolved in ethyl acetate (20 mL) and washed with a solution of 0.1 M HCl (aqueous) (20 mL, three successive washes), then distilled water (20 mL, three successive washes), and finally saturated brine (20 mL, three successive washes; NaCl:water prepared by adding 35 g of NaCl to 100 mL of water). The combined organic fractions were dried over MgSO4 (anhydrous) and then evaporated under reduced pressure to give a yellowish oil (351 mg, quantitative yield).

[0153] Process 3 To a solution of the product from step 2 (50 mg, 0.1 mmol) in a 1:1 mixture of tetrahydrofuran:ethanol (400 μL) was added triethylamine (14 μL, 0.1 mmol), followed by tridecyl acrylate (29 μL, 0.1 mmol). The reaction mixture was stirred overnight at room temperature and then checked for completion by TLC chromatography. The resulting product was purified by column chromatography using a gradient of dichloromethane / dichloromethane:methanol:ammonium hydroxide (80:20:1) (100 / 0 to 0 / 100). Fractions containing the product were combined and evaporated under reduced pressure to give VC-LC-0163 as a yellowish oil (64 mg, 78%).

[0154] VC-LC-0163 was characterized by HPLC-LSD-MS (HPLC Water Alliance ELSD 2424 and X bridge BEH column C8, 4.6 mm, 50 mm, 2.5 μm, gradient: 5% to 95% B (A: water with 0.01% TFA; B: acetonitrile with 0.01% TFA) for 20 min, flow rate = 1 mL / min).

[0155] The injection volume is 2 μL and the temperature in the column is 65°C.

[0156] The detection of the analytes was successfully performed by an evaporative light scattering detector and a simple quadrupole mass detector in positive mode equipped with a spectrophotometer Waters Acquity QDa.

[0157] Retention time: 15.76 min, MS(ES): experimental m / z [M+H] + 754.82{Theoretical m / z[M+H] + 754.61}.

[0158] Example 2: Synthesis of lipid VC-LC-0605 [ka]

[0159] Process 1 2-Oxotetrahydrothiophene-3-carboxylic acid (730 mg, 5 mmol) was dissolved in 13 mL of anhydrous methylene chloride at room temperature. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (870 mg, 4.51 mmol) was then added, followed by dimethylaminopyridine (104 mg, 0.85 mmol) and 2-hexyl-1-decanol (1.03 g, 4.25 mmol). The reaction was stirred overnight at room temperature and then checked for completion by TLC chromatography. The resulting product was purified by flash chromatography using a hexane / ethyl acetate gradient (100 / 0 to 0 / 100). Fractions containing the product were combined and evaporated under reduced pressure to give a yellowish oil (1.33 g, 84%).

[0160] Steps 2 and 3 were carried out as in Example 1 to obtain VC-LC-0605 (41% yield).

[0161] VC-LC-0605 was characterized by HPLC-LSD-MS (HPLC Water Alliance ELSD 2424 and X bridge BEH column C8, 4.6 mm, 50 mm, 2.5 μm, gradient: 5% to 95% B (A: water with 0.01% TFA; B: acetonitrile with 0.01% TFA) for 20 min, flow rate = 1 mL / min).

[0162] The injection volume is 2 μL and the temperature in the column is 65°C.

[0163] The detection of the analytes was successfully performed by an evaporative light scattering detector and a simple quadrupole mass detector in positive mode equipped with a spectrophotometer Waters Acquity QDa.

[0164] Retention time: 16.99 min, MS(ES): Experimental m / z(MH+)769.61{Theoretical m / z[M+H] + 769.79}.

[0165] Example 3: Synthesis of lipid VC-LC-0540 [ka]

[0166] The process of Example 2 was followed to obtain lipid VC-LC-0540, but starting with 5-oxotetrahydrothiophene-3-carboxylic acid instead of 2-oxotetrahydrothiophene-3-carboxylic acid (35% yield).

[0167] VC-LC-0540 was characterized by HPLC-LSD-MS (HPLC Water Alliance ELSD 2424 and X bridge BEH column C8, 4.6 mm, 50 mm, 2.5 μm, gradient: 5% to 95% acetonitrile B (A: water with 0.01% TFA; B: acetonitrile with 0.01% TFA) for 20 min, flow rate = 1 mL / min).

[0168] The injection volume is 2 μL and the temperature in the column is 65°C.

[0169] The detection of the analytes was successfully performed by an evaporative light scattering detector and a simple quadrupole mass detector in positive mode equipped with a spectrophotometer Waters Acquity QDa.

[0170] Retention time: 17.2 min, MS (ES): experimental m / z (MH+) 783.63 {theoretical m / z [M+H] + 783.80}.

[0171] Example 4: Synthesis of lipid VC-LC-0749 [ka]

[0172] Step 1a: DL-homocysteine ​​thiolactone hydrochloride (2172 mg, 14.0 mmol) was dissolved in 24 mL of anhydrous dichloromethane at room temperature. Triethylamine (1961 μL, 14.0 mmol) was then added, followed by EDC hydrochloride (2492 mg, 13 mmol), 4-(dimethylamino)pyridine (247 mg, 2 mmol), and DL-α-lipoic acid (256.42 mg, 10 mmol). The reaction mixture was stirred overnight at room temperature under an argon atmosphere. The crude reaction mixture was then washed twice with distilled water (2 × 40 mL), saturated carbonate solution (40 ml), and finally saturated brine (40 mL). The organic layer was dried over anhydrous MgSO4, filtered, and evaporated under reduced pressure. The resulting residue (intermediate VC-LC-9096) was used without any further purification and characterized by HPLC-ELSD-MS. Retention time: 6.5 min, MS(ES): experimental m / z [M+H] + =306.15. [ka]

[0173] Step 1b: VC-LC-9096 (639 mg, 2.1 mmol) was dissolved in 5 mL of THF and 0.1 mL of HO at room temperature. Tris(2-carboxyethyl)phosphine hydrochloride (611 mg, 2.1 mmol) was then added, followed by octyl acrylate (1120 μL, 5.2 mmol) and triethylamine (1464 μL, 10.5 mmol). The reaction mixture was stirred overnight at room temperature and then analyzed by HPLC-ELSD-MS. The crude reaction mixture was then evaporated under reduced pressure and washed twice with distilled water (2 × 20 mL) and finally with saturated brine (20 mL). The resulting residue was purified by flash chromatography (hexane / ethyl acetate gradient: 100 / 0 to 0 / 100) to give the intermediate compound VC-LC-9097 (65% yield). VC-LC-9097 was characterized by HPLC-ELSD-MS. Retention time: 16.5 min, MS(ES): experimental m / z [M+H] + =676.54. [ka]

[0174] Step 2: Intermediate compound VC-LC-9097 (100 mg, 0.15 mmol) was dissolved in 1.5 mL of anhydrous tetrahydrofuran at room temperature under an argon atmosphere. After complete dissolution of the solid, N,N-diethylethylenediamine (215 μL, 1.5 mmol) was added via syringe. The reaction mixture was stirred at room temperature for 30 minutes and then checked for completion using HPLC-MS. The solvent was then removed under reduced pressure. The dried reaction crude was then redissolved in 80 mL of ethyl acetate and washed three times with 0.1 M HCl solution (1 × 10 mL), water (1 × 10 mL), and finally saturated brine (10 mL). The organic layer was dried over anhydrous MgSO4, filtered, and evaporated under reduced pressure to give intermediate compound VC-LC-9127 as a colorless oil (quantitative yield). VC-LC-9127 was characterized by HPLC-ELSD-MS. Retention time: 13.8 min, MS(ES): experimental m / z [M+H] + =792.66. [ka]

[0175] Step 3: Intermediate compound VC-LC-9127 (120 mg, 0.15 mmol) was dissolved in anhydrous tetrahydrofuran (1.5 mL) under an argon atmosphere. Tetradecyl acrylate (86.0 μL, 0.30 mmol) was then added via syringe. The solution was then stirred at room temperature for 1.5 h (the reaction was monitored using HPLC-MS), after which the solvent was evaporated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane / eluent A: 100 / 0 to 0 / 100, eluent A: dichloromethane / methanol / ammonium hydroxide = 80:20:1 gradient) to give VC-LC-0749 as an oil (83% yield). [ka]

[0176] VC-LC-0749 was characterized by HPLC-ELSD-MS. Retention time: 18.75 min, MS (ES): experimental m / z [M+H] + 1060.96{Theoretical m / z[M+H] + 1060.74}.

[0177] Example 5: Preparation of lipid VC-LC-1194 [ka]

[0178] Step 1: DL-Homocysteine ​​thiolactone hydrochloride (776 mg, 5.0 mmol) was dissolved in 13 mL of methylene chloride at room temperature. Triethylamine (588 μL, 4.3 mmol) was then added, followed by N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (870 mg, 4.5 mmol), dimethylaminopyridine (104 mg, 0.85 mmol), and 2-hexyldecanoic acid (1.30 g, 4.3 mmol). The reaction mixture was stirred overnight at room temperature and then checked for completion by TLC chromatography. The resulting product was purified by flash chromatography using a hexane / ethyl acetate gradient (100 / 0 to 0 / 100). Fractions containing the product were combined and evaporated under reduced pressure to give a yellowish oil (1.39 g, 89%).

[0179] The product was characterized by HPLC-ELSD-MS. Retention time: 14.25 min, MS (ES): experimental m / z [M+H] + 356.44{Theoretical m / z[M+H] + 356,26}. [ka]

[0180] Step 2: The product from Step 1 (249 mg, 0.7 mmol) was dissolved in anhydrous tetrahydrofuran (1.5 mL) and added to a solution of N,N-dimethylethylenediamine (756 μL, 7.0 mmol) in anhydrous tetrahydrofuran (1.0 mL) under an argon atmosphere. The solution was then stirred at room temperature for 30 minutes, after which the solvent was evaporated under reduced pressure. The dried mixture was redissolved in ethyl acetate (20 mL) and washed with a solution of 0.1 M HCl (aqueous) (20 mL, three successive washes), then distilled water (20 mL, three successive washes), and finally saturated brine (20 mL, three successive washes; NaCl:water prepared by adding 35 g of NaCl to 100 mL of water). The combined organic fractions were dried over MgSO (anhydrous) and then evaporated under reduced pressure to give a yellowish oil (quantitative yield).

[0181] The product was characterized by HPLC-LSD-MS. Retention time: 10.5 min, MS(ES): 444.37 experimental m / z [M+H] + :444.55{Theoretical m / z[M+H] +}.

[0182] Step 3: Synthesis of Acrylate. Methylidenebutanedioic acid (250 mg, 1.9 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (940 mg, 4.81 mmol) were dissolved in 12 mL of anhydrous dichloromethane at room temperature. 3,7-Dimethyl-2,6-heptadien-1-ol (845 μL, 4.7 mmol) and triethylamine (932 μL, 6.7 mmol) were then added. The reaction mixture was stirred overnight at room temperature under an argon atmosphere. The crude reaction mixture was then washed twice with distilled water (2 × 10 mL) and finally with saturated brine (10 mL). The organic layer was dried over anhydrous MgSO4, filtered, and evaporated under reduced pressure. The resulting residue was purified by flash chromatography (hexane / ethyl acetate gradient: 100 / 0 to 0 / 100) to give an oil.

[0183] The product was characterized using thin layer chromatography (100% hexane). [ka]

[0184] Step 4: To a solution of the product from Step 2 (35.6 mg, 0.1 mmol) in a 1:1 mixture of tetrahydrofuran:ethanol (400 μL), triethylamine (μL, 0.1 mmol) was added, followed by the product obtained in Step 3 (40.3 mg, 0.1 mmol). The reaction mixture was stirred overnight at room temperature to give the ionized lipid VC-LC-1194, which was confirmed for completion by TLC chromatography. The resulting product was purified by column chromatography using a gradient of dichloromethane / dichloromethane:methanol:ammonium hydroxide (80:20:1) (100 / 0 to 0 / 100). Fractions containing the product were combined and evaporated under reduced pressure to give a yellowish oil (yield: 67%). [ka]

[0185] VC-LC-1194 was characterized by HPLC-ELSD-MS and the detection of the analytes was successfully performed by an evaporative light scattering detector and a simple quadrupole mass detector in positive mode equipped with a spectrophotometer Waters Acquity QDa. Measured retention time: 13.28 min, MS(ES): experimental m / z [M+H] + 846.87{Theoretical m / z[M+H] + 846.64} HPLC-ELSD-MS analysis method (HPLC Water Alliance ELSD 2424 and X-bridge BEH column C8, 4.6 mm, 50 mm, 2.5 μm, gradient 5% to 95% B (A: water containing 100.01% TFA; B: acetonitrile containing 0.01% TFA) in 20 min, flow rate = 1 mL / min). The injection volume was 2 μL, and the temperature in the column was 65 °C. Gradient: (A: water with 0.01% TFA; B: acetonitrile with 0.01% TFA) for 30 min, constant flow rate = 1 mL / min

[0186] Example 6: Preparation and characterization of LNPs containing compounds of the present disclosure

[0187] Encapsulation of RNA into LNPs Encapsulation of mRNA containing the luciferase-encoding sequence of SEQ ID NO: 1 in the 5' to 3' direction into LNPs was performed in some cases by microfluidic technology or by manual means.

[0188] A microfluidic mixer was used, following the same procedure as described in Hassett, KJ et al., "Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines," 2019, Mol. Ther. Nucleic Acids, vol. 15, pp. 1-11. Briefly, purified mRNA was first diluted with sodium citrate buffer at pH 4 and a final concentration of 266 μg / ml. Separately, a mixture of the present ionizable lipids of the present invention: DSPC (Merck 850365P): cholesterol (Sigma C3045): DMG-PEG2000 (Cayman 33945-1) was dissolved in ethanol at a respective molar ratio of 50:10:38.5:1.5 and a lipid-nitrogen:phosphate (N:P) ratio of 5.5:1.

[0189] Alternatively, the LNPs were prepared by the manual method described in Wang X., Liu S., Sun Y. et al. ("Preparation of selective organ-targeting (SORT) lipid nanoparticles (LNPs) using multiple technical methods for tissue-specific mRNA delivery," 2022, Nat. Protoc., doi:10.1038 / s41596-022-00755-x), e.g., manual / bulk mixing. The aqueous mRNA solution was carefully added to the ethanol solution, and the resulting solution was homogenized by pipetting up and down 4–5 times. The resulting LNPs were immediately diluted 1:1 with Tris buffer and dialyzed overnight against Tris buffer containing 15% sucrose. The resulting LNP solution was then collected and the encapsulated mRNA was assessed by Quant-IT® Ribogreen (Invitrogen R11490) according to the manufacturer's instructions.

[0190] The LNP solution was then adjusted to a final mRNA concentration of 100 μg / ml. Particle size distribution, polydispersity, and zeta potential were measured by dynamic light scattering (DLS) using a Malvern Zetasizer Advance Lab Blue Label. RNA encapsulation was assessed by Quant-IT® Ribogreen according to the manufacturer's instructions.

[0191] Finally, the LNP solution was passed through a 0.22 mm filter, and the LNPs were stored at −80° C. until needed.

[0192] Table 6 provides results for several parameters for LNPs containing the specific ionizable lipids shown therein in the following standard formulation: ionizable lipid:non-cationic lipid:sterol:PEG-modified lipid → 50:10:38.5:1.5. [Table 6] TIFF2025538495000143.tif242159TIFF2025538495000144.tif242158TIFF2025538495000145.tif239159 TIFF2025538495000146.tif240159TIFF2025538495000147.tif241159TIFF2025538495000148.tif118159

[0193] As shown in Table 6, each of the tested compounds had an acceptable particle size for the desired purpose. Encapsulation efficiency, measured as the percentage of RNA encapsulated in the LNP, ranged from 100% to approximately 15%. However, this only indicates how easily the nanoparticle components interact with each other to form an LNP capable of encapsulating RNA. It does not indicate intracellular transfection efficiency. Intracellular transfection efficiency of mRNA (encoding expression of luciferase) is measured as luminescence, expressed as total flux (p / s). Total flux is a bioluminescence measurement in photons per second or the average radiance at each pixel integrated over the region of interest.

[0194] As an illustration, LNPs containing compound VC-LC-0550 show an encapsulation efficiency of 26.96%, but the measured total flux is 3.4 × 10 7 p / s, which means that approximately 27% of the RNA was encapsulated when it came into contact with the components of the LNP. The observed total flux values ​​indicated that the LNPs were not degraded in vivo immediately after injection, and that cell transfection was indeed very successful, thus ensuring that the contents encapsulated within the LNPs effectively reached the cytosol after endosomal escape.

[0195] Example 7: Protein expression in mice administered intramuscularly with selected examples of LNPs prepared with lipids of the present invention and containing mRNA as an active ingredient

[0196] Administration of mRNA (LNP) to mice Upon arrival at the experimental facility, female BALB / c mice (Charles River Laboratories), 8–10 weeks old and weighing 18–23 g, were allowed to acclimate to the new conditions for 3–7 days under a 12-h light / dark cycle at a room temperature of 20–24°C, humidity of 50–70%, and light intensity of 60 lux.

[0197] For firefly luciferase activity measurements in mice, LNPs prepared as described above (ionizable lipid:helper lipid:sterol:PEG-modified lipid → 50:10:38.5:1.5) and containing 1 μg of the indicated mRNA in a final volume of 30 μL were injected intramuscularly.

[0198] Starting 4–72 hours after RNA-LNP inoculation, mice were anesthetized by inhalation with 4% isoflurane using a vaporizer. Anesthesia was maintained with 1.5% isoflurane. D-luciferin (Quimigen, 12507) was then injected intraperitoneally at 150 mg / kg, typically approximately 200 μL of a 15 mg / mL stock in PBS for a 20 g mouse. Ten minutes after luciferin inoculation, luciferase images were acquired using an IVIS Lumina XRMS Imaging system according to the manufacturer's instructions.

[0199] Table 7 (A-C) below shows examples of selected LNPs containing ionizable lipids of formula (I) with the same polar head (morpholino) and different R2 and R3 combinations to demonstrate that the general structure of formula (I) produces the desired effect regardless of the alkyl residue. Figure 1 shows the protein expression in mice of the compounds shown in Table 7 (A-C). [Table 7] TIFF2025538495000150.tif242148

[0200] Table 8 shows examples of selected LNPs containing ionizable lipids of formula (I) with various R3 substituents (polar heads) and different combinations of R2 and R3 to demonstrate that the general structure of formula (I) and compounds of formula (II) in particular produce the desired effect regardless of the substituents R1 and R2 and the selected polar head (R3). [Table 8] JPEG2025538495000152.jpg235159JPEG2025538495000153.jpg219159JPEG2025538495000154.j pg229159JPEG2025538495000155.jpg221159JPEG2025538495000156.jpg241159JPEG20255384950 00157.jpg229159JPEG2025538495000158.jpg227159JPEG2025538495000159.jpg232159JPEG202 5538495000160.jpg220159JPEG2025538495000161.jpg235159JPEG2025538495000162.jpg235159

[0201] Tables 7A-C and Table 8 demonstrate that LNPs containing ionizable lipids of Formula II effectively encapsulate polynucleotides and result in high levels of cell transfection in vivo after administration.

[0202] Selected examples of LNPs containing ionizable lipids of formula (III) or ionizable lipids of formula (IV) and encapsulating mRNA for luciferase were also tested in vivo, and the results are shown in Table 9. [Table 9] TIFF2025538495000164.tif236159TIFF2025538495000165.tif241159TIFF2025538495000166.tif78159

[0203] The results summarized in Table 9 demonstrate that LNPs containing ionizable lipids of Formula III and Formula IV effectively encapsulate polynucleotides and result in high levels of cell transfection in vivo after administration.

[0204] Example 8: Immunoassay in mice

[0205] Administration of mRNA-LNPs to mice in different formulations Upon arrival at the experimental facility, female BALB / c mice (Charles River Laboratories), 8–10 weeks old and weighing 18–23 g, were allowed to acclimate to the new conditions for 3–7 days under a 12-h light / dark cycle at a room temperature of 20–24°C, humidity of 50–70%, and light intensity of 60 lux.

[0206] For serum IgG-RBD measurements in mice, therapeutically suitable compositions comprising LNPs prepared as described above (ionizable lipid:helper lipid:sterol:conjugated or PEG-modified lipid) and containing 1 μg of the indicated mRNA of SEQ ID NO: 2 in a final volume of 30 μL were injected intramuscularly. Formulations are summarized in Table 10. Encapsulation of mRNA containing a sequence encoding SEQ ID NO: 2 in a 5' to 3' orientation was performed as previously disclosed in this disclosure. [Table 10]

[0207] Blood was collected (submandibular bleed) on day 21 and blood samples were centrifuged at 10,000 rpm for 10 minutes for serum antibody assay by ELISA, the results of which are summarized in Table 11 below. [Table 11]

[0208] In all cases, the assays carried out to measure serum antibodies by ELISA gave positive results. Samples IM-3, IM-4, IM-5 and IM-6 formulated with ionizable lipids of formula (I), in particular VC-LC-0474, VC-LC-0729, VC-LC-0743 and VC-LC-0867, achieved better results than Control 1 and Control 2, which correspond to the commercial vaccine formulations.

[0209] In each case, a second assay was performed to confirm the efficacy of the booster, administered independently to the mice with the exact same formulation as the previous one, 21 days after the initial administration.

[0210] Blood samples were taken from the submandibular (facial) vein on day 21. Blood samples on day 42 were obtained from euthanized mice.

[0211] Mice were euthanized by trained personnel using appropriate techniques, equipment, and agents. The method chosen was carbon dioxide (CO2) inhalation in a chamber, which complies with the European Union Directive 2010 / 63 / UE on the care and use of laboratory animals in research. The study protocol was approved by the Animal Research Ethics Committee of the University of Zaragoza.

[0212] The results are shown in Table 12 below. [Table 12]

[0213] As summarized in the table, in all cases formulations comprising the ionizable lipid of formula (I) described in this disclosure yielded significantly higher results in serum IgG-RBD assays, with S-specific IFNγ producing cells IM-1, IM-2, IM-3 yielding results comparable to Control 1 and Control 2, and IM-4, IM-5 and IM-6 achieving very good results.

[0214] The World Health Organization (WHO) description of SEQ ID NO: 2 (CAS number: 2417899-77-3, messenger RNA encoding the full-length SARS-CoV-2 spike glycoprotein) is provided below.

[0215] CAP / 5'-UTR / sig / S protein_mut / 3'-UTR / poly(A) UTR = untranslated region; sig = extended signal sequence of S glycoprotein; S protein_mut = S glycoprotein sequence including mutations K986P and V987P; poly(A) = polyadenylation signal tail [ka] [Table 13]

[0216] SEQ ID NO: 2 [ka] TIFF2025538495000173.tif237159Ψ=1-methyl-3'-pseudouridylyl

[0217] Example 9: Comparative Example [ka]

[0218] The compound presented herein, VC-LC-0588, was synthesized via a thiolactone ring-opening reaction following the same synthesis previously reported by Molla et al. (Bioconjugate Chem. 2018, 29, 4, 992-999, referred to as T18U-PY12-A) in a combinatorial library of 288 lipidoids for gene delivery applications, yielding a compound containing an unsaturated bond and a reducible disulfide bond. This study presents results of the compound tested in vitro using HEK293T cells without the addition of helper lipids. The authors reported highly stable liposomes with low toxicity and approximately 95% transfection efficiency. However, this study did not report in vivo experiments in mammals. Therefore, VC-LC-0588 was also studied in vivo herein to compare the differences between compounds of general formula (I) containing a thioether group instead of a disulfide group and the effect of at least one ester group near the S atom in the thioether. As shown in Figure 2B, no transfection (luminescence) was observed in vivo after intramuscular injection of LNPs containing VC-LC-0588 as the ionizable lipid and encapsulating mRNA encoding luciferase in the following formulation: ionizable lipid:helper lipid:sterol:PEG lipid → 50:10:38.5:1.5.

[0219] The compounds shown in Table 13 were also synthesized, characterized, and tested in vivo. These compounds are partially similar to the dimers of formula (I), but have a disulfide moiety introduced, and therefore have the general formula shown here: [ka] (wherein A, B, n, R1 and R3 are the same as defined in general formula (I)). Shows. [Table 14]

[0220] As shown in Figure 2A, no transfection (luminescence) was observable in vivo after intramuscular injection of LNPs containing VC-LC-0236 as the ionizable lipid and encapsulating mRNA encoding luciferase in the following formulation: ionizable lipid:helper lipid:sterol:PEG lipid → 50:10:38.5:1.5.

[0221] Example 10: Optimization of the formulation

[0222] Compound VC-LC-0431 was selected to test different composition ratios.

[0223] As previously shown in Tables 7A-C and Table 8 (Example 7), a standard formulation having the following composition ratio of ionizable lipid:helper lipid:sterol:PEG lipid → 50:10:38.5:1.5 was tested in vivo for the ionizable lipid of formula (I) of the present invention.

[0224] Here, the ionizable lipid is VC-LC-0431, the helper lipid is DOPE, the sterol is cholesterol, and the PEG lipid is DMG-PEG2000, as previously described.

[0225] Table 14 below shows the results obtained for the standard compositions. [Table 15]

[0226] Optimization of this LNP formulation To increase the transfection rate in vivo, different ratios of components in the LNP-like mixture were tested to obtain an optimized formulation.

[0227] The tested compositions shown in Table 15 were injected intraperitoneally into mice, and significant improvements were observed for some of the formulations. [Table 16]

[0228] The results shown in Table 15 demonstrate that it is possible to increase in vivo transfection rates by varying the composition ratio of components in LNP formulations containing the ionizable lipid of Formula (I) of the present invention. LNPs containing compositions 3-8 achieved higher in vivo transfection rates when compared to the previous LNP containing the standard composition of 50:10:38.5:1.5 ionizable lipid:DOPE:cholesterol:DMG-PEG2000. Figure 3 shows in vivo transfection in mice after administration of the eight compositions defined in Table 15.

[0229] Example 11: Optimized formulation with high sterol percentage

[0230] Optimized compositions with a high sterol content and no helper lipids (especially phospholipids) were tested using two different ionizable lipids in the LNP-like mixture, and these formulations were used to test in vivo transfection rates.

[0231] The tested compositions, summarized in Table 16 below, were injected intraperitoneally into mice, and significantly higher transfection efficiencies (measured as total flux) were observed. [Table 17]

[0232] Example 12: Stability of LNPs over time

[0233] Liquid samples stored at T=4°C Two LNPs were tested to determine their stability. The LNPs contained ionizable lipids (Formula I):non-cationic lipids:sterols:PEG-modified lipids in a ratio of 50:10:38.5:1.5, and mRNA containing the luciferase-encoding sequence of SEQ ID NO: 1 in a 5' to 3' orientation. Encapsulation was performed as previously described in this disclosure.

[0234] Formulation A, containing VC-LC-0163 as the ionizable lipid, was administered to mice, and transfection efficiency was measured using a fresh preparation from the first experiment and a preparation stored at T=4°C for 83 days.

[0235] Mice were administered Formulation B, which contained VC-LC-0263 as the ionizable lipid, and transfection efficiency was measured using fresh preparations from the first experiment and preparations stored at T=4° C. for 61 days. The results are summarized in Table 17 below. [Table 18]

[0236] The results obtained demonstrate that LNPs comprising the ionizable lipid of formula (I) of the present disclosure are still able to effect transfection in vivo after extended storage periods under normal refrigerated conditions at 4°C.

[0237] Effect of storage conditions on in vivo immunological studies: Comparative fresh solution stored at -80°C vs. lyophilized formulation stored at 4°C and 25°C To determine whether storage conditions affect in vivo immunity, two different types of LNPs encapsulating mRNA were tested. Thus, a commercially available LNP (LNP A) was compared with another LNP (LNP B) formulated with an ionizable lipid of formula (I) described in this disclosure (specifically, the ionizable lipid VC-LC-0729, 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate). LNP A and LNP B were tested in vivo (in mice) after storage in solution at T=−80°C or after lyophilization at temperatures of 4°C in one case and 25°C in the other. Both LNP A and LNP B were prepared by standard microfluidic mixing as disclosed in Hassett, KJ et al., as defined earlier in this disclosure. Lyophilization of the LNPs can be performed by one skilled in the art using standard methods known in the art. For example, a useful protocol is described in Muramatsu et al. ("Lyophilization provides long-term stability for a lipid nanoparticle-formulated, nucleoside-modified mRNA vaccine" Mol Ther. 2022 May 4;30(5):1941-1951. doi:10.1016 / j.ymthe.2022.02.001).

[0238] For serum IgG-RBD measurements in mice, therapeutically suitable compositions containing LNP A or B prepared as described above (ionized lipid:helper lipid:sterol:conjugated or PEG-modified lipid) and containing 1 μg of the indicated mRNA in a final volume of 30 μL were injected intramuscularly. 21 days after administration, blood was collected by submandibular bleed for serum antibody assay by ELISA, and the samples were centrifuged at 10,000 rpm and t=10 min; the results are shown in Table 18 below.

[0239] The compositions of LNP A and LNP B are described here. LNP A contains the mRNA sequence of SEQ ID NO: 2 encapsulated in a lipid formulation with a ratio of ALC-0315:DSPC:cholesterol:ALC-0159 = 46.3:9.4:42.7:1.6 (% mol). ALC-0315 and ALC-0159 are commercially available (ALC-0315; CAS No.: 2036272-55-4 and ALC-0159; CAS No.: 1849616-42-7). LNP B contains the mRNA sequence of SEQ ID NO:3 encapsulated in a lipid formulation of VC-LC-0729:DOPE:cholesterol:DMG-PEG2000=40.7:34.9:23.3:1.2 (% mol).

[0240] Encapsulation of mRNA containing the sequence of SEQ ID NO:2 or SEQ ID NO:3 in the 5' to 3' direction was performed as described previously in this disclosure.

[0241] SEQ ID NO: 3 [ka] TIFF2025538495000182.tif237159TIFF2025538495000183.tif28159Ψ=1-methyl-3'-pseudouridylyl [Table 19]

[0242] LNP A performed better than LNP B at T=0 days when stored in solution at −80° C. However, results obtained for T=90 days were comparable.

[0243] Surprisingly, for lyophilized LNPs stored at 4°C or 25°C for 90 days, LNP B showed significantly better IgG-RBD measurements than LNP A lyophilized and stored under the same conditions.

[0244] Effect of storage conditions and resuspended lyophilized samples on in vivo immunological studies The LNPs were lyophilized according to standard protocols known in the art as described above.

[0245] Immediately after lyophilization, the samples were resuspended in the same buffer, ie Tris (20 mM) + 15% sucrose, supplemented with 5% trehalose, and immediately stored in a refrigerator at T=4°C.

[0246] Immunological studies were performed 24 and 96 hours after sample resuspension. For serum IgG-RBD measurement in mice, mice were intramuscularly injected with therapeutically suitable compositions of LNPs (A and B) containing 1 μg of the indicated mRNA in a final volume of 30 μL. Blood was collected by submandibular bleeding on day 21 to assay serum antibodies by ELISA, and the samples were centrifuged at 10,000 rpm and t=10 min. The results are summarized in Table 19 below. [Table 20]

[0247] Lyophilized LNP B showed better immune results than commercial LNP A either 24 or 96 hours after resuspension, although in both cases the values ​​obtained were higher after 96 hours (this is actually a result of inter-subject variability), and in a side-by-side comparison it is immediately apparent that LNP B yields significantly higher values ​​than LNP A after 24 hours after resuspension (54% higher for LNP B), and the same is true for the results observed after 96 hours after resuspension (38% higher for LNP B).

[0248] Example 13: Effect of different encapsulation methods on transfection efficiency The impact on transfection efficiency of the synthetic methodology followed to obtain the present LNPs was evaluated for several of the ionizable lipids disclosed herein, specifically VC-LC-0729, VC-LC-0732, VC-LC-0733, VC-LC-0741, VC-LC-0743, VC-LC-0796, VC-LC-0797, VC-LC-0809, VC-LC-0811, VC-LC-0866, VC-LC-0867, VC-LC-0868, VC-LC-0872, and VC-LC-0881.

[0249] Administration of two different therapeutic doses of mRNA (LNP) to mice Upon arrival at the experimental facility, female BALB / c mice (Charles River Laboratories), 8–10 weeks old and weighing 18–23 g, were allowed to acclimate to the new conditions for 3–7 days under a 12-h light / dark cycle at a room temperature of 20–24°C, humidity of 50–70%, and light intensity of 60 lux.

[0250] For firefly luciferase activity measurements in mice, LNPs prepared as described above (ionizable lipid:helper lipid:sterol:PEG-modified lipid → 50:10:38.5:1.5) and containing either 1 μg of the indicated mRNA in a final volume of 30 μL or 0.5 μg of the indicated mRNA in a final volume of 30 μL were injected intramuscularly.

[0251] Starting 4–72 hours after RNA-LNP inoculation, mice were anesthetized by inhalation with 4% isoflurane using a vaporizer. Anesthesia was maintained with 1.5% isoflurane. D-luciferin (Quimigen, 12507) was then injected intraperitoneally at 150 mg / kg, typically approximately 200 μL of a 15 mg / mL stock in PBS for a 20 g mouse. Ten minutes after luciferin inoculation, luciferase images were acquired using an IVIS Lumina XRMS Imaging system according to the manufacturer's instructions.

[0252] In the primary screen, we evaluated in vivo expression of luciferase mRNA encapsulated in LNPs formulated with different ionizable lipids in a standard composition with the following molar ratios (ionizable lipid / helper lipid / sterol / PEG lipid): 50 / 10 / 38.5 / 1.5. LNPs were manually synthesized in 96-well plates.

[0253] In a secondary screen, expression of luciferase mRNA encapsulated in LNPs formulated with different ionizable lipids in a standard composition with the following molar ratios (ionizable lipid / helper lipid / sterol / PEG lipid): 50 / 10 / 38.5 / 1.5 was evaluated. LNPs were again synthesized manually in 96-well plates to ensure consistency of results, and were also synthesized by microfluidics on an Ignite instrument (Precision Nanosystems). Duplicate measurements were performed on the microfluidic LNPs, the results of which are shown in Table 20 below. [Table 21] JPEG2025538495000187.jpg240159JPEG2025538495000188.jpg234159JPEG2025538495000189.jpg52159

[0254] The results obtained in the primary screen showed very good in vivo transfection efficiency results, measured as a total flux as high as 3.61E+08 (for 1 μg of mRNA in a total volume of 30 μL), for example in the case of LNPs containing VC-LC-0729 as the ionizable lipid.

[0255] Secondary screening generally improved the transfection efficiency observed in mice, e.g., values ​​of 1.37E+09 for VC-LC-0729 or 1.34E+09 for VC-LC-0743 (at 1 μg of mRNA in a 30 μL total volume).

[0256] Example 14: Effect of injection dose on transfection efficiency in vivo For VC-LC-0866, VC-LC-0867, VC-LC-0868, VC-LC-0872 and VC-LC-0881, two different doses (1 μg mRNA in a total volume of 30 μL vs. 0.5 μg mRNA in a total volume of 30 μL) were tested in primary and secondary screens in 96-well plates.

[0257] Results obtained in the primary screen at full and half doses, shown in Table 21 below, were comparable for VC-LC-0866 and VC-LC-0868, slightly lower for VC-LC-0867, and improvements were observed for VC-LC-0872 and VC-LC-0881. [Table 22]

[0258] The results shown in Table 21 demonstrate that even when half doses were injected into mice, the observed transfection efficiencies (measured as total flux) were very high, ranging from 1.06E+08 to 2.02E+08. Thus, the ionizable lipids of the present invention contained in LNPs containing therapeutic agents such as mRNA can successfully achieve high transfection rates even at lower doses.

[0259] The present invention includes the following provisions:

[0260] First set of clauses

[0261] 1. Ionizable lipids of formula (II): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof (In the formula, A is [ka] and B is [ka] and D is [ka] and n, m, and p are independently 0, 1, 2, 3, 4, 5, or 6; q is selected from 1, 2 and 3; t is selected from 0, 1 and 2; and wherein q+t=1, 2, or 3, with the proviso that when q+t=1, if R1 is a linear alkyl substituent, then R1 contains at least 7 carbon atoms; R1 and R2 are independently a linear or branched C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R1 and R2 are optionally substituted with one or more substituents selected from the group consisting of -OH, -COOR4, and -C(=O)S-R4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl; and R3 is a heterocycle containing at least one N atom, or alternatively R3 is [ka] and wherein Ra and Rb are independently a straight or branched C1-C6 alkyl optionally substituted with a hydroxyl group.

[0262] 2. The ionizable lipid of clause 1, wherein n is selected from 0, 1, 2 or 3; p is selected from 0, 1, 2 or 3; and m is selected from 0, 1, 2 or 3.

[0263] 3. q is selected from 1 and 2, t is selected from 0 and 1, and where q+t=2, or Or alternatively q is selected from 1 and 2, t is selected from 0 and 1, q+t=2, and R3 is a 5- or 6-membered ring containing one N atom and optionally a second heteroatom preferably selected from N and O, or Or alternatively q is selected from 1 and 2, t is selected from 0 and 1, q+t=2, and R3 has the following structure: [ka] 3. The ionizable lipid according to clause 1 or 2, selected from:

[0264] 4. An ionizable lipid according to any of the preceding clauses, wherein n is selected from 0, 1, 2 or 3, p is selected from 0, 1, 2 or 3, m is selected from 0, 1, 2 or 3, t is 0 or 1, q is 1 or 2, and t+q=2.

[0265] 5. n is 2 or 3, m=t=0, p=q=2, A=B= [ka] and During the ceremony, R1 is a C10-C20 branched alkyl; R2 is a C2-C30 linear or branched alkyl or a C6-C24 linear alkenyl or alkynyl; and R3 is [ka] and In the formula, Ra=Rb=C1-C4 alkyl, Or alternatively, n=2 and R2 is a C2-C30 linear or branched alkyl and R3 is [ka] and In the formula, Ra=Rb=C1-C4 alcohol terminal alkyl, Or alternatively n is 2 or 3, m=t=0, p=q=2, A is [ka] and R1 is a C10-C20 branched alkyl, and R2 is a C3-C30 linear or branched alkyl, and R3 is an imidazole ring; or Or alternatively n=3, m=t=0, p=q=2, A is [ka] and R1 is a C10-C20 branched alkyl, R2 is a C4-C20 linear or branched alkyl, and R3 is a pyrrolidine ring; or Or alternatively n=2, m=t=0, p=q=2, A is [ka] and R1 is a C10-C20 linear or branched alkyl, and R2 is a C4-C20 linear or branched alkyl, and R3 is N-4-methylpiperazine; or Or alternatively n is selected from 2 or 3; m=t=0 and p=q=2, A=B= [ka] and R1 is a C10-C22 linear or branched alkyl, alkenyl, or alkynyl, R2 is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R3 is a morpholino ring, or Or alternatively n is selected from 2 or 3, m=0, p=2, q=t=1; A is [ka] and B=D, R1 is a C10-C22 linear or branched alkyl, R2 is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R3 is a morpholino ring; or Or alternatively n=q=p=2, t=m=0, A is [ka] and B is [ka] and D is [ka] and R1 is a C10-C22 branched alkyl; R2 is a C8-C20 straight chain alkenyl or alkynyl or a C3-C28 straight chain alkyl optionally substituted with one or more substituents selected from OH, -COOR4 and -COSR4, wherein R4 is a C1-C6 alkyl, a C2-C6 alkenyl, a C2-C6 alkynyl or a C12-C25 branched chain alkyl; and R3 is -N(CH3)2 or [ka] 5. The compound according to clause 4, wherein

[0266] 6. VC-LC-0099, VC-LC-0101, VC-LC-0163, VC-LC-0168, VC-LC-0169, VC-LC-0170, VC-LC-0178, VC-LC-0183, VC-LC-0194, VC-LC-0196, VC-LC-0198, VC-LC-0199, VC-LC-0200, VC-LC-0201, VC-LC-0202, VC-LC-0207, VC-LC-0209, VC-LC-0211, VC-LC-0213, VC-LC-0214, VC-LC-0215, VC-LC-0216, VC-LC-0241, VC-LC-0256, VC-LC-0258, VC-LC-0261, VC-LC-0262, VC-LC-0263, VC-LC-0268, VC-LC-0269, VC-LC-0289, VC-LC-0294, VC-LC-0296, VC-LC-0297, VC-LC-0298, VC-LC-0299, VC-LC-0300, VC-LC-0301, VC-LC-0302, VC-LC-0304, VC-LC-0306, VC-LC-0307, VC-LC-0353, VC-LC-0355, VC-LC-0356, VC-LC-0362, VC-LC-0366, VC-LC-0367, VC-LC-0369, VC-LC-0370, VC-LC-0389, VC-LC-0428, VC-LC-0429, VC-LC-0430, VC-LC-0431, VC-LC-0439, VC-LC-0440, VC-LC-0441, VC-LC-0442, VC-LC-0443, VC-LC-0444, VC-LC-0473, VC-LC-0474, VC-LC-0475, VC-LC-0477, VC-LC-0478, VC-LC-0480, VC-LC-0487, VC-LC-0488, VC-LC-0489, VC-LC-0490, VC-LC-0491, VC-LC-0492, VC-LC-0504, VC-LC-0505, VC-LC-0507, VC-LC-0508, VC-LC-0509, VC-LC-0510, VC-LC-0515, VC-LC-0521, VC-LC-0524, VC-LC-0525, VC-LC-0531, VC-LC-0539, VC-LC-0540, VC-LC-0541, VC-LC-0542, VC-LC-0543, as defined in this specificationVC-LC-0544, VC-LC-0546, VC-LC-0547, VC-LC-0548, VC-LC-0549, VC-LC-0550, VC-LC-0551, VC-LC-0553, VC-LC-0554, VC-LC- 0556, VC-LC-0557, VC-LC-0558, VC-LC-0559, VC-LC-0606, VC-LC-0607, VC-LC-0608, VC-LC-0609, VC-LC-0610, VC-LC-0729, V 2. The ionizable lipid according to clause 1, which is a compound selected from the group consisting of C-LC-0732, VC-LC-0733, VC-LC-0741, VC-LC-0743, VC-LC-0796, VC-LC-0797, VC-LC-0809, VC-LC-0811, VC-LC-0866, VC-LC-0867, VC-LC-0868, VC-LC-0872 and VC-LC-0881 or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof.

[0267] 7. A lipid nanoparticle comprising an ionized lipid according to any one of clauses 1 to 6.

[0268] 8.i) Optionally, 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 a non-cationic lipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerin) sodium salt (DOPG), sphingomyelin, and mixtures thereof; a non-cationic lipid, optionally DOPE and / or DSPC; ii) optionally a sterol selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof; and optionally a sterol, which is cholesterol; iii) optionally a PEG-modified lipid selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified phosphatidylcholine, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerin, and mixtures thereof; 8. The lipid nanoparticle of clause 7, further comprising:

[0269] 9. The lipid nanoparticles according to any one of clauses 7 to 8, further comprising a non-cationic lipid, a sterol, and a PEG-modified lipid, wherein the ionizable lipid is present in an amount of 25 to 60 mol%, the PEG-modified lipid is present in an amount of 0.1 to 10 mol%, the non-cationic lipid is present in an amount of 10 to 45 mol%, and the sterol is present in an amount of 10 to 40 mol%.

[0270] 10. The lipid nanoparticle according to any one of clauses 7 to 9, further comprising a pharmaceutically active agent.

[0271] 11. The lipid nanoparticle described in clause 10, wherein the pharmaceutically active agent is selected from the group consisting of a polynucleotide, a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide, and an expression vector comprising a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide.

[0272] 12. The polynucleotide is a natural or artificial deoxyribonucleic acid (DNA) or a natural or artificial ribonucleic acid (RNA), Further optionally, the polynucleotide comprises one or more amino acids selected from the group consisting of pseudouridine, N1-methylpseudouridine (also known as 1-methylpseudouridine or m1Ψ), N6-methyladenosine (also known as m6A), 2-thiouridine (also known as s2U), 4'-thiouridine, 5-methylcytosine (also known as 5mC), 2-thio-1-methyl-1-deazapseudouridine, 2-thio-1-methylpseudouridine, 2-thio-5-azauridine, 2-thiodihydropseudouridine, 2-thiodihydrouridine, 2-thiopseudouridine, 4-methoxy-5-azauridine, 2-thiodihydropseudour ... -2-thiopseudouridine, 4-methoxypseudouridine, 4-thio-1-methylpseudouridine, 4-thiopseudouridine, 5-azauridine, dihydropseudouridine, 5-methyluridine (also referred to as m5U), 5-methoxyuridine, 2'-O-methyluridine and combinations thereof, in particular said chemical modification is N1-methylpseudouridine, 5-methoxyuridine or combinations thereof, in particular said chemical modification is N1-methylpseudouridine, Further optionally, the polynucleotide is ribonucleic acid (RNA), Further optionally, the RNA is selected from the group consisting of short interfering RNA (siRNA), self-replicating RNA (srRNA), circular RNA (circRNA), self-amplifying RNA (saRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), small interfering RNA (siRNA), small RNA (sRNA), dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), messenger RNA (mRNA) and mixtures thereof, in particular, the RNA is mRNA; 12. The lipid nanoparticle according to clause 11.

[0273] 13. A pharmaceutical composition comprising a lipid nanoparticle according to any one of clauses 10 to 12 and a pharmaceutically acceptable excipient or carrier.

[0274] 14. For use in a method for treating a disease or disorder in a subject in need thereof, optionally selected from the group consisting of infectious diseases, cancer, proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases and metabolic diseases, comprising administering to the subject a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition, wherein optionally the subject is a human. or for use in a method for inducing an immune response in a subject, for use in a method for therapeutic immunization of a subject, for use as a vaccine, or for use in gene therapy; or for use in the prevention or treatment of COVID-19 The lipid nanoparticles according to any one of clauses 10 to 12 or the pharmaceutical composition according to clause 13.

[0275] 15. Use of lipid nanoparticles according to any one of clauses 7 to 9 as defined herein as an encapsulating agent.

[0276] Second set of clauses

[0277] 1. Ionizable lipids of formula (II): [ka] Formula (II) or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof (In the formula, A is [ka] and B is [ka] and D is [ka] and n, m, and p are independently 0, 1, 2, 3, 4, 5, or 6; q is selected from 1, 2 and 3; t is selected from 0, 1 and 2; and wherein q+t=1, 2, or 3, provided that when q+t=1, R1 contains at least 7 carbon atoms when it is a linear alkyl substituent; R1 and R2 are independently a linear or branched C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R1 and R2 are optionally substituted with one or more substituents selected from the group consisting of -OH, -COOR4, and -C(=O)S-R4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl; and R3 is a heterocycle containing at least one N atom, or alternatively R3 is [ka] and wherein Ra and Rb are independently a straight or branched C1-C6 alkyl optionally substituted with a hydroxyl group.

[0278] 2. The ionizable lipid of clause 1, wherein n is selected from 0, 1, 2 or 3; p is selected from 0, 1, 2 or 3; and m is selected from 0, 1, 2 or 3.

[0279] 3. q is selected from 1 and 2, t is selected from 0 and 1, and where q+t=2, or Or alternatively q is selected from 1 and 2, t is selected from 0 and 1, q+t=2, and R3 is a 5- or 6-membered ring containing one N atom and optionally a second heteroatom preferably selected from N and O, or Or alternatively q is selected from 1 and 2, t is selected from 0 and 1, q+t=2, and R3 has the following structure: [ka] 3. The ionizable lipid according to clause 1 or 2, selected from:

[0280] 4. An ionizable lipid according to any of the preceding clauses, wherein n is selected from 0, 1, 2 or 3, p is selected from 0, 1, 2 or 3, m is selected from 0, 1, 2 or 3, t is 0 or 1, q is 1 or 2, and t+q=2.

[0281] 5. n is 2 or 3, m=t=0, p=q=2, A=B= [ka] and R1 is a C10-C20 branched alkyl; R2 is a C2-C30 linear or branched alkyl or a C6-C24 linear alkenyl or alkynyl; and R3 is [ka] and In the formula, Ra=Rb=C1-C4 alkyl, Or alternatively, n=2 and R2 is a C2-C30 linear or branched alkyl and R3 is [ka] and In the formula, Ra=Rb=C1-C4 alcohol terminal alkyl, Or alternatively n is 2 or 3, m=t=0, p=q=2, A is [ka] and R1 is a C10-C20 branched alkyl, and R2 is a C3-C30 linear or branched alkyl, and R3 is an imidazole ring; or Or alternatively n=3, m=t=0, p=q=2, A is [ka] and R1 is a C10-C20 branched alkyl, R2 is a C4-C20 linear or branched alkyl, and R3 is a pyrrolidine ring; or Or alternatively n=2, m=t=0, p=q=2, A is [ka] and R1 is a C10-C20 linear or branched alkyl, and R2 is a C4-C20 linear or branched alkyl, and R3 is N-4-methylpiperazine; or Or alternatively n is selected from 2 or 3; m=t=0 and p=q=2, A=B= [ka] and R1 is a C10-C22 linear or branched alkyl, alkenyl, or alkynyl, R2 is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R3 is a morpholino ring, or Or alternatively n is selected from 2 or 3, m=0, p=2, q=t=1; A is [ka] and B=D, R1 is a C10-C22 linear or branched alkyl, R2 is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R3 is a morpholino ring; or Or alternatively n=q=p=2, t=m=0, A is [ka] and B is [ka] and D is [ka] and R1 is a C10-C22 branched alkyl; R2 is a C8-C20 straight chain alkenyl or alkynyl or a C3-C28 straight chain alkyl optionally substituted with one or more substituents selected from OH, -COOR4 and -COSR4, wherein R4 is a C1-C6 alkyl, a C2-C6 alkenyl, a C2-C6 alkynyl or a C12-C25 branched chain alkyl; and R3 is -N(CH3)2 or [ka] 5. The compound according to clause 4, wherein

[0282] 6. VC-LC-0099, VC-LC-0101, VC-LC-0163, VC-LC-0168, VC-LC-0169, VC-LC-0170, VC-LC-0178, VC-LC-0183, VC-LC-0194, VC-LC-0196, VC-LC-0198, VC-LC-0199, VC-LC-0200, VC-LC-0201, VC-LC-0202, VC-LC-0207, VC-LC-0209, VC-LC-0211, VC-LC-0213, VC-LC-0214, VC-LC-0215, VC-LC-0216, VC-LC-0241, VC-LC-0256, VC-LC-0258, VC-LC-0261, VC-LC-0262, VC-LC-0263, VC-LC-0268, VC-LC-0269, VC-LC-0289, VC-LC-0294, VC-LC-0296, VC-LC-0297, VC-LC-0298, VC-LC-0299, VC-LC-0300, VC-LC-0301, VC-LC-0302, VC-LC-0304, VC-LC-0306, VC-LC-0307, VC-LC-0353, VC-LC-0355, VC-LC-0356, VC-LC-0362, VC-LC-0366, VC-LC-0367, VC-LC-0369, VC-LC-0370, VC-LC-0389, VC-LC-0428, VC-LC-0429, VC-LC-0430, VC-LC-0431, VC-LC-0439, VC-LC-0440, VC-LC-0441, VC-LC-0442, VC-LC-0443, VC-LC-0444, VC-LC-0473, VC-LC-0474, VC-LC-0475, VC-LC-0477, VC-LC-0478, VC-LC-0480, VC-LC-0487, VC-LC-0488, VC-LC-0489, VC-LC-0490, VC-LC-0491, VC-LC-0492, VC-LC-0504, VC-LC-0505, VC-LC-0507, VC-LC-0508, VC-LC-0509, VC-LC-0510, VC-LC-0515, VC-LC-0521, VC-LC-0524, VC-LC-0525, VC-LC-0531, VC-LC-0539, VC-LC-0540, VC-LC-0541, VC-LC-0542, VC-LC-0543, as defined in this specificationVC-LC-0544, VC-LC-0546, VC-LC-0547, VC-LC-0548, VC-LC-0549, VC-LC-0550, VC-LC-0551, VC-LC-0553, VC-LC-0554, VC-LC- 0556, VC-LC-0557, VC-LC-0558, VC-LC-0559, VC-LC-0606, VC-LC-0607, VC-LC-0608, VC-LC-0609, VC-LC-0610, VC-LC-0729, V 2. The ionizable lipid according to clause 1, which is a compound selected from the group consisting of C-LC-0732, VC-LC-0733, VC-LC-0741, VC-LC-0743, VC-LC-0796, VC-LC-0797, VC-LC-0809, VC-LC-0811, VC-LC-0866, VC-LC-0867, VC-LC-0868, VC-LC-0872 and VC-LC-0881 or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof.

[0283] 7. A lipid nanoparticle comprising an ionized lipid according to any one of clauses 1 to 6.

[0284] 8.i) Optionally, 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 a non-cationic lipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerin) sodium salt (DOPG), sphingomyelin, and mixtures thereof; a non-cationic lipid, optionally DOPE and / or DSPC; ii) optionally a sterol selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof; and optionally a sterol, which is cholesterol; iii) optionally a PEG-modified lipid selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified phosphatidylcholine, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerin, and mixtures thereof; 8. The lipid nanoparticle of clause 7, further comprising:

[0285] 9. The lipid nanoparticles according to any one of clauses 7 to 8, further comprising a non-cationic lipid, a sterol, and a PEG-modified lipid, wherein the ionizable lipid is present in an amount of 25 to 60 mol%, the PEG-modified lipid is present in an amount of 0.1 to 10 mol%, the non-cationic lipid is present in an amount of 10 to 45 mol%, and the sterol is present in an amount of 10 to 40 mol%.

[0286] 10. The lipid nanoparticle according to any one of clauses 7 to 9, further comprising a pharmaceutically active agent.

[0287] 11. The lipid nanoparticle described in clause 10, wherein the pharmaceutically active agent is selected from the group consisting of a polynucleotide, a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide, and an expression vector comprising a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide.

[0288] 12. The polynucleotide is a natural or artificial deoxyribonucleic acid (DNA) or a natural or artificial ribonucleic acid (RNA), Further optionally, the polynucleotide comprises one or more amino acids selected from the group consisting of pseudouridine, N1-methylpseudouridine (also known as 1-methylpseudouridine or m1Ψ), N6-methyladenosine (also known as m6A), 2-thiouridine (also known as s2U), 4'-thiouridine, 5-methylcytosine (also known as 5mC), 2-thio-1-methyl-1-deazapseudouridine, 2-thio-1-methylpseudouridine, 2-thio-5-azauridine, 2-thiodihydropseudouridine, 2-thiodihydrouridine, 2-thiopseudouridine, 4-methoxy-5-azauridine, 2-thiodihydropseudour ... -2-thiopseudouridine, 4-methoxypseudouridine, 4-thio-1-methylpseudouridine, 4-thiopseudouridine, 5-azauridine, dihydropseudouridine, 5-methyluridine (also referred to as m5U), 5-methoxyuridine, 2'-O-methyluridine and combinations thereof, in particular said chemical modification is N1-methylpseudouridine, 5-methoxyuridine or combinations thereof, in particular said chemical modification is N1-methylpseudouridine, Further optionally, the polynucleotide is ribonucleic acid (RNA), Further optionally, the RNA is selected from the group consisting of short interfering RNA (siRNA), self-replicating RNA (srRNA), circular RNA (circRNA), self-amplifying RNA (saRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), small interfering RNA (siRNA), small RNA (sRNA), dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), messenger RNA (mRNA) and mixtures thereof, in particular, the RNA is mRNA; 12. The lipid nanoparticle according to clause 11.

[0289] 13. A pharmaceutical composition comprising a lipid nanoparticle according to any one of clauses 10 to 12 and a pharmaceutically acceptable excipient or carrier.

[0290] 14. For use in a method for treating a disease or disorder in a subject in need thereof, optionally selected from the group consisting of infectious diseases, cancer, proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases and metabolic diseases, comprising administering to the subject a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition, wherein optionally the subject is a human. or for use in a method for inducing an immune response in a subject, for use in a method for therapeutic immunization of a subject, for use as a vaccine, or for use in gene therapy; or for use in the prevention or treatment of COVID-19 The lipid nanoparticles according to any one of clauses 10 to 12 or the pharmaceutical composition according to clause 13.

[0291] 15. Use of lipid nanoparticles according to any one of clauses 7 to 9 as defined herein as an encapsulating agent.

[0292] Third set of clauses

[0293] 1. Ionizable lipids of formula (I): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof (In the formula, A is selected from the group consisting of -NR'-C(O)-, -C(O)-NR'-, -C(O)-O-, and -OC(O)-, where R' is selected from the group consisting of H, methyl, and ethyl; B is selected from the group consisting of -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -OC(O)-; n, m, and p are independently selected from 0, 1, 2, 3, 4, 5, and 6; q is selected from 1, 2 and 3; t is selected from 0, 1 and 2; and where q+t=1, 2 or 3, R3 is a heterocycle containing at least one N atom, or alternatively R3 is [ka] and wherein Ra and Rb are independently a linear or branched C1-C6 alkyl optionally substituted with a hydroxyl group; X is -D-R2, [ka] is selected from wherein j, j', and j'' are independently selected from 0, 1, and 2; Z is -R1 and [ka] is selected from wherein f and f' are independently selected from 0, 1, 2, 3, 4, 5, and 6; g and g' are independently selected from 1, 2, 3, 4, 5 and 6; D is independently selected from —C(O)—O— or —OC(O)—; and wherein R1, R'1, R2, R'2, and R''2 are independently a straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R1, R'1, R2, R'2, and R''2 are optionally substituted with one or more substituents selected from the group consisting of -OH, -COOR4, and -C(=O)SR4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl.

[0294] 2. When q+t=1, X=-D-R2, D=-OC(O)-, A=-NH-C(O)-, B=-NH-C(O)-, Z=R1, and R1 is a linear or branched alkyl, the moiety -(CH2) m 10. The ionizable lipid of clause 1, wherein R1 comprises at least 7 carbon atoms.

[0295] 3. n = t = m = 0, q = p = 1, X is [ka] and wherein one of j and j′ is 0 and the other is 1; When D is -OC(O)-, A=-C(O)-NH-, B=-NH-C(O)-, Z is R1, and R1 is C1-C18 alkyl, 10. The ionizable lipid of claim 1, wherein R2 and R'2 are different from C7-C18 alkyl.

[0296] 4. Compound of formula (II): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A, B, D, n, m, p, q, t, q+t, R1 and R2 are as defined in clause 1; or Or alternatively When q+t=1, X=-D-R2, D=-OC(O)-, A=-NH-C(O)-, B=-NH-C(O)-, Z=R1, and R1 is a linear or branched alkyl, the moiety -(CH2) m -R1 contains at least 7 carbon atoms; 1. The ionizable lipid according to clause 1.

[0297] 5. The ionizable lipid of clause 4, wherein A is -NH-C(O)- or -C(O)-NH-.

[0298] 6. The ionizable lipid of clause 4 or 5, wherein n is selected from 0, 1, 2 and 3; p is selected from 0, 1, 2 and 3; and m is selected from 0, 1, 2 and 3.

[0299] 7. The ionizable lipid of clauses 4-6, wherein q is selected from 1 and 2, and t is selected from 0 and 1, and wherein q+t=2.

[0300] 8. The ionizable lipid according to any one of clauses 1 to 7, wherein R3 is a 5- or 6-membered ring containing one N atom and optionally a second heteroatom preferably selected from N and O.

[0301] 9. R3 has the structure: [ka] 9. The ionizable lipid according to any one of clauses 1 to 8, selected from:

[0302] 10. The ionizable lipid according to any one of clauses 4 to 9, wherein n is selected from 0, 1, 2 or 3, p is selected from 0, 1, 2 or 3, m is selected from 0, 1, 2 or 3, t is 0 or 1, q is 1 or 2, and t+q=2.

[0303] 11. n is 2 or 3, m = t = 0, p = q = 2, A=B= [ka] and R1 is a C10-C20 branched alkyl; R2 is a C2-C30 linear or branched alkyl or a C6-C24 linear alkenyl or alkynyl; and R3 is [ka] and 11. The compound according to clause 10, wherein Ra=Rb=C1-C4 alkyl.

[0304] 12. n=2, and R2 is a C2-C30 linear or branched alkyl, and R3 is [ka] and 11. An ionizable lipid according to clause 10, wherein Ra = Rb = C1-C4 alcohol terminal alkyl.

[0305] 13. n is 2 or 3, m = t = 0, p = q = 2, A is [ka] and 11. The compound according to clause 10, wherein R1 is a C10-C20 branched alkyl, and R2 is a C3-C30 straight or branched alkyl, and R3 is an imidazole ring.

[0306] 14. n = 3, m = t = 0, p = q = 2, A is [ka] and 11. The ionizable lipid of clause 10, wherein R1 is a C10-C20 branched alkyl, R2 is a C4-C20 straight or branched alkyl, and R3 is a pyrrolidine ring.

[0307] 15. n = 2, m = t = 0, p = q = 2, A is [ka] and 11. The compound according to clause 10, wherein R1 is a C10-C20 straight or branched chain alkyl, and R2 is a C4-C20 straight or branched chain alkyl, and R3 is N-4-methylpiperazine.

[0308] 16.n is chosen from 2 or 3, m=t=0 and p=q=2, A=B= [ka] and 11. The ionizable lipid of clause 10, wherein R1 is a C10-C22 linear or branched alkyl, alkenyl, or alkynyl, R2 is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R3 is a morpholino ring.

[0309] 17. n is selected from 2 or 3, m = 0, p = 2, q = t = 1, A is [ka] and 11. The ionizable lipid of clause 10, wherein B=D, R1 is a C10-C22 linear or branched alkyl, R2 is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R3 is a morpholino ring.

[0310] 18. n = q = p = 2, t = m = 0, A is [ka] and B is [ka] and D is [ka] and R1 is a C10-C22 branched alkyl; R2 is a C8-C20 linear alkenyl or alkynyl or a C3-C28 linear alkyl optionally substituted with one or more substituents selected from OH, -COOR4 and -COSR4, and R4 is a C1-C6 alkyl, a C2-C6 alkenyl, a C2-C6 alkynyl or a C12-C25 branched alkyl; and R3 is -N(CH3)2 or [ka] 11. The ionizable lipid of clause 10, wherein

[0311] 19. Compound of formula (III): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof, wherein: A is —NH—C(O)— or —C(O)—NH—; R1, R2, and R'2 are independently selected from linear or branched C1-C30 alkyl, C2-C30 alkenyl, and C2-C30 alkynyl, wherein each of R1, R2, and R'2 is optionally substituted with one or more substituents selected from the group consisting of -OH, -COO(R4), and -COSR4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl; and B, D, n, m, p, q, t, q+t, j, j' and R3 are as defined in clause 1; or Or alternatively n=t=m=0, q=p=1, A=B=-NH-C(O)-, 2. The ionizable lipid of claim 1, wherein when one of j and j' is 0 and the other is 1, and D=-OC(O)-, and R1 is selected from C1-C18 alkyl, then R2 and R'2 are not C7-C18 alkyl.

[0312] 20. The compound is a compound of formula (IV): [ka] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A is —NH—C(O)— or —C(O)—NH—; B, D, n, m, p, q, t, q+t, f, f', g, g' and R3 are as defined in clause 1; 1. The ionizable lipid according to clause 1.

[0313] 21. VC-LC-0099, VC-LC-0101, VC-LC-0163, VC-LC-0168, VC-LC-0169, VC-LC-0170, VC-LC-0178, VC-LC-0183, VC-LC-0194, VC-LC-0196, VC-LC-0198, VC-LC-0199, VC-LC-0200, VC-LC-0201, VC-LC-0202, VC-LC-0207, VC-LC-0209, VC-LC-0211, VC-LC-0213, VC-LC-0214, VC-LC-0215, VC-LC-0216, VC-LC-0241, VC-LC-0256, VC-LC-0258, VC-LC-0261, VC-LC-0262, VC-LC-0263, VC-LC-0268, VC-LC-0269, VC-LC-0289, VC-LC-0294, VC-LC-0296, VC-LC-0297, VC-LC-0298, VC-LC-0299, VC-LC-0300, VC-LC-0301, VC-LC-0302, VC-LC-0304, VC-LC-0306, VC-LC-0307, VC-LC-0353, VC-LC-0355, VC-LC-0356, VC-LC-0362, VC-LC-0366, VC-LC-0367, VC-LC-0369, VC-LC-0370, VC-LC-0389, VC-LC-0418, VC-LC-0428, VC-LC-0429, VC-LC-0430, VC-LC-0431, VC-LC-0439, VC-LC-0440, VC-LC-0441, VC-LC-0442, VC-LC-0443, VC-LC-0444, VC-LC-0473, VC-LC-0474, VC-LC-0475, VC-LC-0477, VC-LC-0478, VC-LC-0480, VC-LC-0487, VC-LC-0488, VC-LC-0489, VC-LC-0490, VC-LC-0491, VC-LC-0492, VC-LC-0504, VC-LC-0505, VC-LC-0507, VC-LC-0508, VC-LC-0509, VC-LC-0510, VC-LC-0515, VC-LC-0521, VC-LC-0524, VC-LC-0525, VC-LC-0531, VC-LC-0539, VC-LC-0540, VC-LC-0541, VC-LC-0542 described in this specificationVC-LC-0543, VC-LC-0544, VC-LC-0546, VC-LC-0547, VC-LC-0548, VC-LC-0549, VC-LC-0550, VC-LC-0551, VC-LC-0553, VC-LC-0554, VC-LC-0556, VC-L C-0557, VC-LC-0558, VC-LC-0559, VC-LC-0606, VC-LC-0607, VC-LC-0608, VC-LC-0609, VC-LC-0610, VC-LC-0729, VC-LC-0730, VC-LC-0731, VC-LC-07 32, VC-LC-0733, VC-LC-0734, VC-LC-0735, VC-LC-0736, VC-LC-0737, VC-LC-0738, VC-LC-0739, VC-LC-0741, VC-LC-0742, VC-LC-0743, VC-LC-0757, V C-LC-0796、VC-LC-0797、VC-LC-0798、VC-LC-0799、VC-LC-0800、VC-LC-08 01、VC-LC-0802、VC-LC-0803、VC-LC-0804、VC-LC-0805、VC-LC-0806、VC-LC -0807、VC-LC-0809、VC-LC-0810、VC-LC-0811、VC-LC-0828、VC-LC-0848、VC-LC-0864、VC-LC-0865、VC-LC-0866、VC-LC-0867、VC-LC-0868、VC-LC-086 9, VC-LC-0870, VC-LC-0871, VC-LC-0872, VC-LC-0873, VC-LC-0876, VC-LC-0877, VC-LC-0880, VC-LC-0881, VC-LC-0890, VC-LC-0895, VC-LC-0919, VC -LC-0921、VC-LC-0922、VC-LC-0936、VC-LC-0940、VC-LC-0944、VC-LC-094 5、VC-LC-0949、VC-LC-0953、VC-LC-0973、VC-LC-1065、VC-LC-1068、VC-LC- 1082, VC-LC-1083, VC-LC-1084, VC-LC-1085, VC-LC-1088, VC-LC-1089, VC-LC-1090, VC-LC-1091, VC-LC-1094, VC-LC-1095, VC-LC-1096, VC-LC-10972. The ionizable lipid according to clause 1, which is a compound selected from the group consisting of VC-LC-1098, VC-LC-1105, VC-LC-1111, VC-LC-1143, VC-LC-1192, VC-LC-1202, VC-LC-1203, VC-LC-1224, VC-LC-1230, VC-LC-1254 or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof.

[0314] 22. Lipid nanoparticles comprising an ionized lipid according to any one of clauses 1 to 21.

[0315] 23. Optionally, 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 23. The lipid nanoparticle of clause 22, further comprising a non-cationic lipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sphingomyelin and mixtures thereof, optionally wherein the non-cationic lipid is DOPE and / or DSPC.

[0316] 24. Optionally, a sterol selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof; 24. The lipid nanoparticle of clause 22 or 23, further comprising a sterol or steroid precursor, optionally cholesterol.

[0317] 25. The lipid nanoparticle according to any one of clauses 22 to 23, further comprising a PEG-modified lipid selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified phosphatidylcholine, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerin and mixtures thereof, or alternatively a conjugated lipid.

[0318] 26. A composition comprising a non-cationic lipid, a sterol, and a PEG-modified lipid, wherein the ionizable lipid is present in an amount of 25 to 60 mol%, the PEG-modified lipid is present in an amount of 0.1 to 10 mol%, the non-cationic lipid is present in an amount of 10 to 45 mol%, and the sterol is present in an amount of 10 to 40 mol%; or Or alternatively The ionized lipid is present in an amount of 25 to 64 mol%, the PEG-modified lipid is present in an amount of 0.1 to 1.5 mol%, and the sterol is present in an amount of 35 to 74 mol%. 25. The lipid nanoparticle according to any one of clauses 22 to 24.

[0319] 27. The lipid nanoparticle according to any one of clauses 22 to 26, further comprising a pharmaceutically active agent.

[0320] 28. The lipid nanoparticle described in clause 27, wherein the pharmaceutically active agent is selected from the group consisting of a polynucleotide, a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide, and an expression vector comprising a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide.

[0321] 29. The pharmaceutically active agent is a polynucleotide; Optionally, the polynucleotide is a natural or artificial deoxyribonucleic acid (DNA) or ribonucleic acid (RNA); Further optionally, the polynucleotide comprises one or more of the following amino acids: pseudouridine, N1-methylpseudouridine (also referred to as 1-methylpseudouridine or m1Ψ), N6-methyladenosine (also referred to as m6A), 2-thiouridine (also referred to as s2U), 4'-thiouridine, 5-methylcytosine (also referred to as 5mC), 2-thio-1-methyl-1-deazapseudouridine, 2-thio-1-methylpseudouridine, 2-thio-5-azauridine, 2-thiodihydropseudouridine, 2-thiodihydrouridine, 2-thiopseudouridine, 4-methoxamic acid, 4-methyl-1-methyl-2 ... and at least one chemical modification selected from the group consisting of 2-thiopseudouridine, 4-methoxypseudouridine, 4-thio-1-methylpseudouridine, 4-thiopseudouridine, 5-azauridine, dihydropseudouridine, 5-methyluridine (also referred to as m5U), 5-methoxyuridine, 2'-O-methyluridine, and combinations thereof, in particular the chemical modification is N1-methylpseudouridine, 5-methoxyuridine, or combinations thereof, in particular the chemical modification is N1-methylpseudouridine. Further optionally, the polynucleotide is ribonucleic acid (RNA), Further optionally, the RNA is selected from the group consisting of short interfering RNA (siRNA), self-replicating RNA (srRNA), circular RNA (circRNA), self-amplifying RNA (saRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), small interfering RNA (siRNA), small RNA (sRNA), dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), messenger RNA (mRNA) and mixtures thereof, in particular, the RNA is mRNA; 29. The lipid nanoparticle according to clause 28.

[0322] 30. Lipid nanoparticles according to clause 29, wherein the ionizable lipid:RNA ratio (N / P) in the LNP is in the range of 20:1 to 2:1, in particular 10:1 to 3:1.

[0323] 31. A pharmaceutical composition comprising lipid nanoparticles according to any one of clauses 21 to 24 and a pharmaceutically acceptable excipient or carrier.

[0324] 32. A pharmaceutical composition according to clause 31 which is a vaccine and optionally further comprises an adjuvant.

[0325] 33. A method for treating a disease or disorder in a subject in need thereof, optionally selected from the group consisting of an infectious disease, cancer, a proliferative disease, a genetic disease, an autoimmune disease, diabetes, a neurodegenerative disease, a cardiovascular disease, a renal vascular disease, and a metabolic disease, comprising administering to the subject a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition, Further optionally, for use in the method, wherein the subject is a human, or for use in a method for inducing an immune response in a subject, for use in a method for therapeutic immunization of a subject, for use as a vaccine, or for use in gene therapy; or for use in the prevention or treatment of COVID-19 Lipid nanoparticles according to any one of clauses 23 to 26 or pharmaceutical composition according to clause 31 or 32.

[0326] 34. Use of lipid nanoparticles according to any one of clauses 22 to 33 as defined herein as an encapsulating agent.

[0327] Prior art documents 1. Molla MR. et al. (See Molla MR. et al. "One-Pot Parallel Synthesis of Lipid Library via Thiolactone Ring Opening and Screening for Gene Delivery." Bioconjug Chem. 2018, vol. 29(4), pp. 992-999. doi:10.1021 / acs.bioconjchem.8b00007) 2. Churusova, S. et al. (2021). "Palladium(II) Pincer Complexes of Functionalized Amides with S-Modified Cysteine ​​and Homocysteine ​​Residues: Cytotoxic Activity and Different Aspects of Their Biological Effect on Living Cells." Inorganic Chemistry. 2021, vol. 60, pp. 9880-9898. 3. U.S. Patent No. 4,929,736 2. Garbiras, BJ; Marburg, S. "Preparation of Carboxythiolactones and Their Active Derivatives." Synthesis, 1999, vol. 2, pp. 270-274; doi:10.1055 / s-1999-3377. 3. Hassett, KJ et al., "Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines," 2019, Mol. Ther. Nucleic Acid, vol. 15, pp. 1-11, DOI: 10.1016 / j.omtn.2019.01.013. 4. Wang X., Liu S., Sun Y., et al., "Preparation of selective organ-targeting (SORT) lipid nanoparticles (LNPs) using multiple technical methods for tissue-specific mRNA delivery," Nat. Protoc., 2022, doi:10.1038 / s41596-022-00755-x

Claims

1. Ionizable lipids of formula (I): 【Chemistry 1】 or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof (In the formula, A is selected from the group consisting of —NR′—C(O)—, —C(O)—NR′—, —C(O)—O—, and —O—C(O)—, where R′ is selected from the group consisting of H, methyl, and ethyl; B is selected from the group consisting of —NH—C(O)—, —C(O)—NH—, —C(O)—O—, and —O—C(O)—; n, m, and p are independently selected from 0, 1, 2, 3, 4, 5, and 6; q is selected from 1, 2 and 3; t is selected from 0, 1 and 2; and where q+t=1, 2, or 3; R ３ is a heterocycle containing at least one N atom, or alternatively R ３ teeth 【Chemistry 2】 and wherein Ra and Rb are independently a linear or branched C1-C6 alkyl optionally substituted with a hydroxyl group; X is -D-R ２ , 【Transformation 3】 is selected from wherein j, j′, and j″ are independently selected from 0, 1, and 2; Z is -R １ and 【Chemistry 4】 is selected from wherein f and f′ are independently selected from 0, 1, 2, 3, 4, 5, and 6; g and g' are independently selected from 1, 2, 3, 4, 5 and 6; D is independently selected from —C(O)—O— or —O—C(O)—; And R １ , R' １ , R ２ , R' ２ and R'' ２ are independently straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R １ , R' １ , R ２ , R' ２ and R'' ２ -OH, -COOR ４ and -C(=O)SR ４ and optionally substituted with one or more substituents selected from the group consisting of: ４ is C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl, however, ・q+t=1, X=-DR ２ , D=-OC(O)-, A=-NH-C(O)-, B=-NH-C(O)-, Z=R １ and R １ When is a linear or branched alkyl, the moiety -(CH ２ ) ｍ -R １ contains at least 7 carbon atoms, and the following compound S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-methyl homocysteate: 【Transformation 5】 S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-L-cysteic acid methyl ester: 【Transformation 6】 and (L)-CH ３ OOC-CH ２ SCH ２ -CH(COOCH ３ )-NH-CO-imidazole is excluded).

2. n=t=m=0, q=p=1, X is 【Transformation 7】 and wherein one of j and j′ is 0 and the other is 1; D is —O—C(O)—, A=—NH—C(O)—, B=—NH—C(O)—, and Z is R １ and R １ is C1-C18 alkyl, R ２ and R' ２ is different from C7-C18 alkyl, The ionized lipid of claim 1.

3. Compound of formula (II): 【Transformation 8】 or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; In the formula, A, B, D, n, m, p, q, t, q+t, R １ and R ２ is as defined in claim 1, however, ・q+t=1, X=-DR ２ , D=-OC(O)-, A=-NH-C(O)-, B=-NH-C(O)-, Z=R １ and R １ When is a linear or branched alkyl, the moiety -(CH ２ ) ｍ -R １ contains at least 7 carbon atoms, the following compound S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-methyl homocysteate: 【Chemistry 9】 S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-L-cysteic acid methyl ester: 【Chemistry 10】 and (L)-CH ３ OOC-CH ２ SCH ２ -CH(COOCH ３ )-NH-CO-imidazole is excluded; The ionized lipid of claim 1.

4. 4. The ionizable lipid of claim 3, wherein A is —NH—C(O)— or —C(O)—NH—.

5. 5. The ionizable lipid of claim 3 or 4, wherein n is selected from 0, 1, 2 and 3, p is selected from 0, 1, 2 and 3, and m is selected from 0, 1, 2 and 3.

6. q is selected from 1 and 2, t is selected from 0 and 1, and q+t=2; or Or alternatively q is selected from 1 and 2, t is selected from 0 and 1, q+t=2, and R ３ is a 5- or 6-membered ring containing one N atom and optionally a second heteroatom preferably selected from N and O, Or alternatively q is selected from 1 and 2, t is selected from 0 and 1, q+t=2, and R ３ has the following structure: 【Chemistry 11】 The ionizable lipid according to claims 3 to 5, selected from:

7. n is selected from 0, 1, 2 or 3, p is selected from 0, 1, 2 or 3, m is selected from 0, 1, 2 or 3, t is 0 or 1, q is 1 or 2, and t + q = 2. An ionizable lipid according to any one of claims 3 to 6.

8. n is 2 or 3, m=t=0, p=q=2, A=B=—NH—C(O)— or —C(O)—NH—; R １ is a C10-C20 branched alkyl; R ２ is a C2-C30 straight or branched alkyl or a C6-C24 straight alkenyl or alkynyl, and R ３ teeth 【Chemistry 12】 and wherein Ra=Rb=C1-C4 alkyl; Or alternatively, n=2 and R ２ is a C2 to C30 linear or branched alkyl, and R ３ teeth 【Chemistry 13】 and In the formula, Ra=Rb=C1-C4 alcohol terminal alkyl, Or alternatively n is 2 or 3, m=t=0, p=q=2, A is —NH—C(O)— or —C(O)—NH—; R １ is a C10-C20 branched alkyl, and R ２ is a C3 to C30 linear or branched alkyl, and R ３ is an imidazole ring, or Or alternatively n=3, m=t=0, p=q=2, A is —NH—C(O)— or —C(O)—NH—; R １ is a C10-C20 branched alkyl, and R ２ is a C4 to C20 linear or branched alkyl, and R ３ is a pyrrolidine ring, or Or alternatively n=2, m=t=0, p=q=2, A is —NH—C(O)— or —C(O)—NH—; R １ is a C10-C20 linear or branched alkyl, and R ２ is a C4 to C20 linear or branched alkyl, and R ３ is N-4-methylpiperazine, or Or alternatively n is selected from 2 or 3; m=t=0 and p=q=2; A=B=—NH—C(O)— or —C(O)—NH—; R １ is a C10-C22 linear or branched alkyl, alkenyl, or alkynyl; R ２ is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R ３ is a morpholino ring, or Or alternatively n is selected from 2 or 3, m=0, p=2, q=t=1; A is —NH—C(O)— or —C(O)—NH—; B=D and R １ is a C10 to C22 linear or branched alkyl; R ２ is a C3-C30 linear or branched alkyl or a C3-C30 linear alkenyl or alkynyl, and R ３ is a morpholino ring, or Or alternatively n=q=p=2, t=m=0, A is 【Chemistry 14】 and B is 【Chemistry 15】 and D is 【Chemistry 16】 and R １ is a C10-C22 branched alkyl; R ２ is a C8-C20 linear alkenyl or alkynyl, or OH, —COOR ４ and -COSR ４ wherein R is a C3 to C28 linear alkyl optionally substituted with one or more substituents selected from ４ is C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl or C12-C25 branched alkyl; And R ３ HA-N(CH ３ ) ２ or 【Chemistry 17】 8. The compound of claim 7, wherein:

9. The compound is a compound of formula (III): [Chemistry 18] or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A is —NH—C(O)— or —C(O)—NH—; R １ , R ２ and R' ２ are independently selected from linear or branched C1-C30 alkyl, C2-C30 alkenyl, and C2-C30 alkynyl, wherein R １ , R ２ , R' ２ are —OH, —COO(R ４ ) and -COSR ４ and optionally substituted with one or more substituents selected from the group consisting of: ４ is C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl, and B, D, n, m, p, q, t, q+t, j, j' and R ３ is as defined in claim 1 or Or alternatively n=t=m=0, q=p=1, A=B=-NH-C(O)-, one of j and j' is 0 and the other is 1, and D=-O-C(O)-, and R １ is C1-C18 alkyl, R ２ and R' ２ is not a C7-C18 alkyl; The ionized lipid of claim 1.

10. The compound is a compound of formula (IV): 【Chemistry 19】 or a pharmaceutically acceptable salt thereof or any one stereoisomer thereof; During the ceremony, A is —NH—C(O)— or —C(O)—NH—, and B, D, n, m, p, q, t, q+t, f, f', g, g' and R ３ is as defined in claim 1, The ionized lipid of claim 1.

11. Tridecyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0099), 8-methylnonyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0101), 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propane as defined herein Tridecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0163), tetradecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0168), octadecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0169), hexyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propanoate hexadecyl (VC-LC-0170), tridecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0178), tetradecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0183), tetradecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)tridecyl propanoate (VC-LC-0194), 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate 8-methylnonyl (VC-LC-0196), 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0198),Tetradecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0199), octadecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0200), 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamide )-4-oxobutyl)thio)hexadecyl propanoate (VC-LC-0201), 6-methylheptyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0202), octyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0207), tridecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0209), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0211), dodecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0213), tetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0214), octadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0215), hexadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0216),Tridecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((3-(pyrrolidin-1-yl)propyl)amino)butyl)thio)propanoate (VC-LC-0241), Tridecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0256), 8-methylno 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoic acid 8-methylnonyl ester (VC-LC-0261), 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoic acid octadecyl ester (VC-LC-0262), 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4 hexadecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0263), hexyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0268), octyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0269), 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3 -oleamido-4-oxobutyl)thio)tridecyl propanoate (VC-LC-0289), 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butanoate heptadecan-9-yl (VC-LC-0294), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate heptadecan-9-yl (VC-LC-0296),4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate heptadecan-9-yl (VC-LC-0297), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate heptadecan-9-yl (VC-LC-0298), 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-( Heptadecan-9-yl ((3-oxo-3-(tetradecyloxy)propyl)thio)methyl)butanoate (VC-LC-0299), heptadecan-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0300), heptadecan-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate -yl (VC-LC-0301), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate heptadecan-9-yl (VC-LC-0302), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate heptadecan-9-yl (VC-LC-0304), 4-((3-(1H-imidazol-1-yl)propyl heptadecan-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0306), heptadecan-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0307), 2-decyltetradecyl 3-((4-((3-morpholinopropyl)amino)-3-oleamido-4-oxobutyl)thio)propanoate (VC-LC-0353),Octadec-9-en-1-yl 3-((4-((3-morpholinopropyl)amino)-3-oleamido-4-oxobutyl)thio)propanoate (VC-LC-0355), 2-decyltetradecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-oleamido-4-oxobutyl)thio)propanoate (VC-LC-0356), heptadecyl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoate Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-(dodecyloxy)-3-oxopropyl)thio)butanoate (VC-LC-0362), Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-(dodecyloxy)-3-oxopropyl)thio)butanoate (VC-LC-0366), Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butanoate (VC-LC-0367), Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butanoate (VC-LC-0368), Heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)butanoate (VC-LC-0369), heptadecan-9-yl 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)butanoate (VC-LC-0370), 2-decyl 3-((4-((3-morpholinopropyl)amino)-4-oxo-3-stearamidobutyl)thio)propanoate tetradecyl (VC-LC-0389), 2-hexyldecyl 6-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-6-oxohexanoate (VC-LC-0418), 2-decyltetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0428),Dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0429), Octadec-9-ene-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0429), -yl (VC-LC-0430), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0431), 4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl, Heptadecan-9-yl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0439), heptadecan-9-yl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0440), heptadecan-9-yl 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoate (VC-LC-0441), 4-((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)-2- ((1-methylpiperidin-4-yl)carbamoyl)heptadecan-9-yl butanoate (VC-LC-0442), (Z)-2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)heptadecan-9-yl butanoate (VC-LC-0443), 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)heptadecan-9-yl butanoate (VC-LC-0444), 3-( Tridecyl (4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0473), 2-hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0474), 8-methylnonyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0475) ), dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0477), tetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0478), hexadecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0480),2-Decyltetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0487), dec-2-yn-1-yl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0488), octadec-9-ene 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate -1-yl (VC-LC-0489), tridecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0490), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0491), 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)- 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0492), 2-decyltetradecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0504), dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propanoate (VC-LC-0505), tridecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0507), 2-hexyldecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0508), 8-methylnonyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0509),2-Ethylhexyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0510), 6-methylheptyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0515), 6-methylheptyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0516), 2-decyltetradecyl 4-((1-methylpiperidin-4-yl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butanoate (VC-LC-0524), 2-hexyldecyl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutanoate (VC-LC-0525), 2-(((3-( 2-Hexyldecyl (Z)-4-((1-methylpiperidin-4-yl)amino)-2-(((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0531), 2-Hexyldecyl (Z)-4-((1-methylpiperidin-4-yl)amino)-2-(((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0539), 4-((3-morpholinopropyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyl) heptadecan-9-yl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0540), heptadecan-9-yl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0541), heptadecan-9-yl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0542),2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0543), 2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0544), 4-((3-morpholinopropyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)thio)methyl)-4-oxopropyl Heptadecan-9-yl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0546), heptadecan-9-yl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0547), heptadecan-9-yl 2-(((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0548). , 2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0549), 2-(((3-(hexyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0550), 4-((3-morpholinopropyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl 2-(((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0551), 2-(((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0553), 2-(((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate heptadecan-9-yl (VC-LC-0554),Tridecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0556), 2-hexyldecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0557), 8-hydroxybenzoic acid esters of 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate -methylnonyl (VC-LC-0558), 2-ethylhexyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0559), 2-hexyldecyl 4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0606), 4-((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)-2-((3- 2-Hexyldecyl 4-((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0607), 2-Hexyldecyl 4-((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0608), 2-Hexyldecyl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0609), 2-((3-morpholinopropyl)carbamoyl)-4- 2-hexyldecyl ((3-oxo-3-(tetradecyloxy)propyl)thio)butanoate (VC-LC-0610), 2-hexyldecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0639), 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0729),8-methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0730), 2-ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0731), dodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0732), tetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0733), 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyl, Octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0734), hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0735), 6-methylheptyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0736), 3-((4-((2-(dimethylamino)ethyl) docosyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0737), hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0738), octyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0739), )ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate 2-decyltetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0741), dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0742), octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0743), octyl 8-(2-((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)ethyl)-3-ethyl-15-((3-(octyloxy)-3-oxopropyl)thio)-7,10-dioxo-18-thia-3,6,9-triaza-21-heneicosylate (VC-LC-0757), tridecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0796),2-Hexyldecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0797), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0798), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0799), E) 2-ethylhexyl propanoate (VC-LC-0799), dodecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0800), tetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0801), 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0802), octadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0802), hexadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0803), 6-methylheptyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0804), docosyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0805), hexyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0806), octyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0807),2-Decyltetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0809), dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoate (VC-LC-0810), 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl )thio)propanoic acid octadec-9-en-1-yl (VC-LC-0811), 3-((4-(4-(2-hydroxyethyl)piperazin-1-yl)-3-(2-octyldodecanamido)-4-oxobutyl)thio)tridecyl (VC-LC-0828), 3-((3-(2-octyldodecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propanoic acid 8-methylnonyl (VC-LC-0848), 3-((4-((2-(dimethylamino)ethyl)amino) )-3-(2-octyldodecanamido)-4-oxobutyl)thio)tridecyl propanoate (VC-LC-0864), 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0865), 8-methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0866), 2-ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0867), dodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0868), tetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0869),Octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0870), hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0871), 6-methyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate butyl (VC-LC-0872), docosyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0873), hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0876), 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate Octyl propanoate (VC-LC-0877), dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0880), octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0881), 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)propanoate (VC-LC-0882), 6-methylheptyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0890), octyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0895), 2-hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0919),2-Ethylhexyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0921), dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0922), tridecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-09 36), 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0937), dodecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0940), 6-(3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate -methylheptyl (VC-LC-0944), docosyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0945), octyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (VC-LC-0949), 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propanoate (Z)-octadec-9-en-1-yl 6-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-6-oxohexanoate (VC-LC-0973), 2-octyldodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1065),7-(2-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)ethyl)-2-methyl-14-((3-(octyloxy)-3-oxopropyl)thio)-6,9-dioxo-17-thia-2,5,8-triaza-20-eicosanoic acid octyl ester (VC-LC-1068), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-1-oxo-4-((3-oxo-3-(tridecyl dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl)dipropionate (VC-LC-1082), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl), Dioctyl bis(sulfanediyl))dipropionate (VC-LC-1083), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1084), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-(( Dioctyl 3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1085), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(octadecyloxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionic acid dioctyl (VC-LC-1088), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(hexadecyloxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1089), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1089), dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1090), dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1091),3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((2-decyltetradecyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1094), 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1094), dioctyl 3,3'-((8-oxo-8-((1-oxo-4-((3-oxo-3-(tridecyloxy)propyl)thio)-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)octane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1096); dioctyl 3,3'-((8-oxo-8-((1-oxo-4-((3-oxo-3-(tridecyloxy)propyl)thio)-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)octane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1096) ), 3,3'-((8-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1097), 3,3'-((8-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl )amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1098), 3,3'-((8-((4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dioctyl dipropionate (VC-LC-1105), 3,3'-((8-((4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))(Z)-dioctyl dipropionate (VC-LC-1111), 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (9Z,12Z)-octadeca-9,1 2-dien-1-yl (VC-LC-1143), 3,7,11-trimethyldodeca-2,6,10-trien-1-yl (2E,6E)-3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1192), 3,7-dimethylocta-2,6-dien-1-yl bis((E)-2-(((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)methyl)succinate) ( VC-LC-1194), tetradecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1202), dodecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1203), 2-(((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecane bis(3,7-dimethyloctyl) 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)methyl)succinate (VC-LC-1218), 2-hexyldecyl 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate (VC-LC-1224), (9Z,12Z)-octadeca-9, 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propanoate,12-dien-1-yl (VC-LC-1230), 2-((2-(dimethylamino)ethyl)carbamoyl)-4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)butanoic acid 2-hexyldecyl (VC-LC-1254), 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyl)oxy)carbonyl)-4-oxobutyl)thio)methyl)co Bis(3,7-dimethyloctyl) succinate (VC-LC-1281), bis((E)-3,7-dimethylocta-2,6-dien-1-yl) 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyl)oxy)carbonyl)-4-oxobutyl)thio)methyl) succinate (VC-LC-1282), bis(((4-((2-butyloctyl)oxy)-3-((2-(dimethylamino)ethyl)amino) bis((E)-3,7-dimethylocta-2,6-dien-1-yl)2-((4-((2-decyltetradecyl)oxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1291), bis(3,7-dimethyloctyl)2-(((4-((2-decyltetradecyl)oxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1295), The ionizable lipid according to claim 1, which is a compound selected from the group consisting of bis((E)-3,7-dimethylocta-2,6-dien-1-yl) 2-(((4-((2-decyltetradecyl)oxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1296) or a pharmaceutically acceptable salt thereof, or any one stereoisomer thereof.

12. Ionizable lipids of formula (I): 【Chemistry 20】 (In the formula, A is selected from the group consisting of —NR′—C(O)—, —C(O)—NR′—, —C(O)—O—, and —O—C(O)—, where R′ is selected from the group consisting of H, methyl, and ethyl; B is selected from the group consisting of —NH—C(O)—, —C(O)—NH—, —C(O)—O—, and —O—C(O)—; n, m, and p are independently selected from 0, 1, 2, 3, 4, 5, and 6; q is selected from 1, 2 and 3; t is selected from 0, 1 and 2; and where q+t=1, 2, or 3; R ３ is a heterocycle containing at least one N atom, or alternatively R ３ teeth 【Chemistry 21】 and wherein Ra and Rb are independently a linear or branched C1-C6 alkyl optionally substituted with a hydroxyl group; X is -D-R ２ , 【Chemistry 22】 is selected from wherein j, j′, and j″ are independently selected from 0, 1, and 2; Z is -R １ and 【Chemistry 23】 is selected from wherein f and f′ are independently selected from 0, 1, 2, 3, 4, 5, and 6; g and g' are independently selected from 1, 2, 3, 4, 5 and 6; D is independently selected from —C(O)—O— or —O—C(O)—; And R １ , R' １ , R ２ , R' ２ and R'' ２ are independently straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, or C2-C30 alkynyl, wherein R １ , R' １ , R ２ , R' ２ and R'' ２ -OH, -COOR ４ and -C(=O)SR ４ and optionally substituted with one or more substituents selected from the group consisting of: ４ is C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl. or a pharmaceutically acceptable salt thereof, or a lipid nanoparticle comprising any one stereoisomer thereof.

13. i) optionally 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 a non-cationic lipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sphingomyelin, and mixtures thereof; a non-cationic lipid, optionally DOPE and / or DSPC; ii) optionally a sterol selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof; and optionally a sterol, which is cholesterol; iii) a complex lipid, or alternatively optionally a PEG-modified lipid selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified phosphatidylcholine, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerin, and mixtures thereof; The lipid nanoparticle of claim 12, further comprising:

14. The ionizable lipid is present in an amount of 25 to 60 mol %, the PEG-modified lipid or the complex lipid is present in an amount of 0.1 to 10 mol %, the non-cationic lipid is present in an amount of 10 to 45 mol %, and the sterol is present in an amount of 10 to 40 mol %; Or alternatively The ionizable lipid is present in an amount of 25 to 64 mol%, the PEG-modified lipid or the complex lipid is present in an amount of 0.1 to 1.5 mol%, and the sterol is present in an amount of 35 to 74 mol%. The lipid nanoparticles of claim 13.

15. The lipid nanoparticle of any one of claims 12 to 14, further comprising a pharmaceutically active agent.

16. The lipid nanoparticle of claim 15, wherein the pharmaceutically active agent is selected from the group consisting of a polynucleotide, a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide, and an expression vector comprising a DNA construct comprising a promoter operably linked to a sequence encoding the polynucleotide.

17. the polynucleotide is a natural or artificial deoxyribonucleic acid (DNA) or a natural or artificial ribonucleic acid (RNA); Further optionally, the polynucleotide comprises pseudouridine, N1-methylpseudouridine (also referred to as 1-methylpseudouridine or m1Ψ), N6-methyladenosine (also referred to as m6A), 2-thiouridine (also referred to as s2U), 4'-thiouridine, 5-methylcytosine (also referred to as 5mC), 2-thio-1-methyl-1-deazapseudouridine, 2-thio-1-methylpseudouridine, 2-thio-5-azauridine, 2-thiodihydropseudouridine, 2-thiodihydrouridine, 2-thiopseudouridine, 4-methoxy-5-azauridine, 2-thiodihydropseudour ... at least one chemical modification selected from the group consisting of N1-methylpseudouridine, 4-methoxypseudouridine, 4-thio-1-methylpseudouridine, 4-thiopseudouridine, 5-azauridine, dihydropseudouridine, 5-methyluridine (also referred to as m5U), 5-methoxyuridine, 2'-O-methyluridine and combinations thereof, in particular said chemical modification is N1-methylpseudouridine, 5-methoxyuridine or combinations thereof, in particular said chemical modification is N1-methylpseudouridine; Further optionally, the polynucleotide is ribonucleic acid (RNA); Further optionally, said RNA is selected from the group consisting of short interfering RNA (siRNA), self-replicating RNA (srRNA), circular RNA (circRNA), self-amplifying RNA (saRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), small interfering RNA (siRNA), small RNA (sRNA), dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), messenger RNA (mRNA) and mixtures thereof, in particular said RNA is mRNA; The lipid nanoparticle of claim 16.

18. A pharmaceutical composition comprising the lipid nanoparticles of any one of claims 12 to 17 and a pharmaceutically acceptable excipient or carrier.

19. For use in a method for treating a disease or disorder in a subject in need thereof, optionally selected from the group consisting of an infectious disease, a cancer, a proliferative disease, a genetic disease, an autoimmune disease, diabetes, a neurodegenerative disease, a cardiovascular disease, a renal vascular disease, and a metabolic disease, comprising administering a therapeutically effective amount of said nanoparticle composition or said pharmaceutical composition to said subject, wherein optionally said subject is a human. or for use in a method for inducing an immune response in a subject, for use in a method for therapeutic immunization of a subject, for use as a vaccine, or for use in gene therapy; or for use in the prevention or treatment of COVID-19 Lipid nanoparticles according to any one of claims 12 to 18 or a pharmaceutical composition according to claim 18.

20. Use of lipid nanoparticles according to any one of claims 12 to 19 as defined herein as an encapsulating agent.

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