Ionizable lipids and lipid nanoparticles containing them

Abstract

The present invention provides an ionizable lipid of formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them; a lipid nanoparticle comprising the ionizable lipid (in particular as an encapsulant), optionally comprising a pharmaceutically active agent; and a pharmaceutical composition comprising the lipid nanoparticles. The present invention also provides a lipid nanoparticle for a drug or a pharmaceutical composition comprising the same, and a use of the lipid nanoparticle as an encapsulant. # imgabs0 #

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of European Patent Application EP22383119.9 filed on November 21, 2022, European Patent Application EP23382067.9 filed on January 27, 2023, and European Patent Application EP23382401.0 filed on April 28, 2023. Technical field

[0003] 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 delivering active ingredients, including polynucleotides, to cells. Background art

[0004] Today, the treatment of different diseases, such as infectious diseases, is changing rapidly, especially because new polynucleotide delivery systems are being designed and their efficiency optimized, making them useful alternatives to traditional therapies.

[0005] Gene delivery systems designed for systemic delivery and intended to encapsulate polynucleotides (e.g., RNA) must be safe and non - toxic, have a nanoscale size, provide protection against degradation for the polynucleotide, remain intact in the system for a given period of time to reach their target, and be easily degraded once the payload is released.

[0006] The use of RNA in gene therapy and vaccination is generally considered safer than the use of DNA because RNA does not involve the risk of stable integration into the genome of the transfected cell. In addition, RNA is more easily degraded in vivo, so the risk of generating unwanted anti - RNA antibodies (which would reduce the efficacy of the therapy and produce very serious side effects) is lower. Two main limitations of RNA - based therapies are low transfection rates and limited protein production efficiency. To compensate for these limitations, the dose is increased to achieve the desired therapeutic effect. Therefore, higher doses also mean increased costs and some adverse side effects observed.

[0007] One of the more effective intracellular delivery techniques for encapsulating and delivering active molecules, such as genetic material (e.g., mRNA) in vaccines, is lipid nanoparticles (LNPs). Currently, the LNPs used in commercial mRNA vaccines typically contain four types of lipids in their composition, namely cationic or ionizable lipids, structural lipids (which are sterols such as cholesterol), PEG - modified lipids, and non - cationic lipids such as phospholipids. In particular, ionizable lipids are crucial in LNPs.

[0008] The properties of ionizable lipids have a great impact on the protection of genetic material encapsulated within LNPs, as they allow the maintenance of the structure and physicochemical properties of said genetic material until the LNP reaches the target (e.g., tissue, cell), where the genetic material is released.

[0009] Ionizable lipids generally exhibit the following general structure:

[0010] Lipophilic moiety ---- Linker group ---- Bridge ---- Hydrophilic moiety

[0011] There are many instances in the prior art where the hydrophilic moiety contains an ionizable amine, a functional group whose formal charge is affected by changes in pH. In addition, the presence of an ester group that is readily hydrolyzed by enzymes once the genetic material is released into the target tissue / cell has been disclosed to promote the degradation of the ionizable lipid, thereby enhancing its biocompatibility and biodegradability. It has also been described that the proximity of the ester group to the hydrophilic moiety has a great impact on the potency of the lipid.

[0012] For example, Moderna uses the SM-102 lipid in its SARS-CoV-2 vaccine formulation (Spikevax), and Pfizer has obtained a license for ALC-0315 (Comirnaty) from Acuitas.

[0013]

[0014] 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 with hydrophobic tails containing reducible disulfide groups and allowing the obtaining of stable liposomes.

[0015] Despite the continuous improvement of such LNP delivery systems, there are still problems with the efficient and specific delivery of targeting agents. Thus, there remains a need for alternative ionizable lipids and / or LNPs that overcome some of the drawbacks of these prior arts, particularly those showing good stability, high transfection efficiency, and safety. Summary of the Invention

[0016] The present inventors have developed a new ionizable lipid that allows the obtaining of lipid nanoparticles (LNPs) that can be effectively used as non-viral vectors for delivering active ingredients including polynucleotides to cells. In particular, the LNPs containing the ionizable lipid of the present disclosure exhibit enhanced / improved transfection rates.

[0017] The present inventors have found that an ionizable lipid of formula (I) as defined hereinafter:

[0018]

[0019] which comprises a polar head (R3), at least one thioether moiety, at least one stereocenter, at least one ester moiety and two moieties A and B selected from amide or ester, wherein A, B, X, Z, R3, m, n, p, q and t are as defined in the present disclosure, is particularly suitable for preparing LNPs capable of encapsulating active agents. Furthermore, the present inventors have found that the in vivo transfection rate of LNPs prepared with the ionizable lipid of the present disclosure and containing an active agent is unexpectedly high compared to other commercial or standard compositions known in the art.

[0020] Therefore, it is evident from the data provided in the examples that the ionizable lipid of the present disclosure provides a new tool that overcomes some of the limitations of known LNPs.

[0021] The higher transfection efficiency of the LNPs of the present disclosure may allow for a reduction in the therapeutic dose of polynucleotides (such as RNA) required in gene therapy and vaccination, which may help to mitigate the associated secondary effects. At the same time, the LNPs of the present disclosure exhibit good thermal and mechanical properties, and it has been demonstrated that their in vivo transfection efficiency remains even after storage at 4 °C for three months without further precautions, which means a long shelf life under normal refrigeration conditions and can be extended at lower temperatures (e.g., at household refrigerator temperatures from -25 °C to -15 °C, or can be frozen at even lower temperatures down to -90 °C).

[0022] Accordingly, a first aspect of the present disclosure relates to an ionizable lipid of formula (I):

[0023]

[0024] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0025] A is selected from -NR'-C(O)-, -C(O)-NR'-, -C(O)-O- and -O-C(O)-, wherein R' is selected from H, methyl and ethyl;

[0026] B is selected from -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -O-C(O)-;

[0027] n, m and p are independently selected from 0, 1, 2, 3, 4, 5 and 6;

[0028] q is selected from 1, 2 and 3;

[0029] t is selected from 0, 1 and 2;

[0030] and wherein q + t = 1, 2 or 3;

[0031] R3 is a heterocycle containing at least one N atom, or R3 is:

[0032]

[0033] wherein Ra and Rb are independently a straight-chain or branched C1-C6 alkyl optionally substituted with a hydroxyl group;

[0034] X is selected from -D-R2,

[0035] wherein j, j’ and j” are independently selected from 0, 1 and 2,

[0036] Z is selected from -R1 and

[0037] wherein f and f’ are independently selected from 0, 1, 2, 3, 4, 5 and 6,

[0038] wherein g and g’ are independently selected from 1, 2, 3, 4, 5 and 6,

[0039] wherein each D is independently selected from -C(O)-O- or -O-C(O)-,

[0040] 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 -OH, -COOR4 and -C(=O)SR4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl;

[0041] provided that

[0042] - when q + t = 1, X = -D-R2, D = -O-C(O)-, A = -NH-C(O)-, B = -NH-C(O)-, Z = R1 and R1 is a straight-chain or branched alkyl, the moiety -(CH2) m - R1 contains at least 7 carbon atoms;

[0043] - Excluding the compound S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-homocysteine methyl ester,

[0044]

[0045] and S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-L-cysteine methyl ester

[0046]

[0047] both of which 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,

[0048] and the compound (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole disclosed in US4929736.

[0049] Another aspect of the present invention relates to an LNP that comprises an ionizable lipid of formula (I) as defined herein, and further comprises an ionizable lipid not claimed above and below.

[0050] In another aspect, the LNP of the present disclosure may further comprise a pharmaceutically active agent and thus can be formulated into a drug together with excipients and carriers.

[0051] Accordingly, another aspect of the present invention relates to a pharmaceutical composition that comprises an LNP containing a pharmaceutically active agent as defined herein and a pharmaceutically acceptable excipient or carrier.

[0052] The LNP or pharmaceutical composition of the present disclosure containing a pharmaceutically active agent can be used for pharmaceutical applications.

[0053] Accordingly, 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, in particular for a method of treating a disease or disorder in a subject in need thereof, or for a method of inducing an immune response in a subject, for a method of therapeutic immunization of a subject, for use as a vaccine or for gene therapy. The method comprises administering to the subject a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition.

[0054] Another aspect of the present invention relates to the use of an LNP as defined herein as an encapsulant for an active ingredient. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 Related to Example 7, protein expression in mice administered LNP is depicted, the LNP being prepared with the lipids of the present invention as follows: VC-LC-0163, VC-LC-0289, VC-LC-0374, VC-LC-0389, VC-LC-0553, VC-LC-0554, VC-LC-0353, VC-LC-0355, and containing RNA as the active ingredient. The tested and depicted lipids contain the same R3 substituent (polar head) in all cases (except for one case) and different combinations of R2 and R3 substituents, showing good transfection results, as confirmed by the total flux measurements for each lipid in the images and Table 7 (A - C).

[0056] Figure 2 Related to Comparative Example 9, in vivo protein expression in mice injected with LNP is depicted, the LNP comprising VC-LC-0236(2A) as the ionizable lipid in the LNP composition and VC-LC-0588(2B) as the ionizable lipid in the LNP composition.

[0057] Figure 3 Related to Example 10, protein expression in mice injected with the LNP of Table 11 is depicted, the LNP having different composition ratios for the ionizable lipid VC-LC-0431, DOPE, cholesterol, and DMG-PEG2000. DETAILED DESCRIPTION

[0058] Unless otherwise indicated, all terms used in this application are to be understood in their ordinary meaning as known in the art. Other more specific definitions of certain terms used in this application are set forth below and are intended to apply uniformly throughout this specification and the claims, unless a more extensive definition is provided by an expressly stated definition.

[0059] As used herein, references to an indefinite number are synonymous with "at least one" or "one or more". Unless otherwise specified, references to an indefinite number as used herein also include plural referents.

[0060] The term "and / or" means that any one of the options associated therewith is possible, or at least two of the options occur simultaneously.

[0061] The term "moiety" refers to a specific segment or functional group of a molecule or compound.

[0062] As used herein, the term "subject" refers to any mammal, including humans and non-human mammals.

[0063] As used herein, the term "C1-C# alkyl" refers to a saturated straight-chain or branched hydrocarbon containing 1 to # carbon atoms and optionally substituted. Non-limiting examples of alkyl 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.

[0064] The term "C2-C# alkenyl" refers to an unsaturated straight-chain or branched hydrocarbon chain containing 2 to # carbon atoms and at least one or more double bonds and optionally substituted. Examples of alkenyl can include, but are not limited to, vinyl (i.e., ethenyl), allyl, propenyl, butenyl, pentenyl, and hexenyl, dodecenyl, tetradecenyl, and hexadecenyl.

[0065] The term "C2-C# alkynyl" refers to an unsaturated straight-chain or branched hydrocarbon chain containing 2 to # carbon atoms and at least one or more triple bonds and optionally substituted. Examples of alkynyl include, but are not limited to, ethynyl, prop-1-ynyl, prop-2-ynyl, 1-methylprop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl.

[0066] The term "heterocycle containing at least one N atom" refers to an optionally mono- or polysubstituted cyclic system containing one or more rings, wherein at least one ring contains at least one nitrogen.

[0067] 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 can be substituted by replacing a hydrogen atom with a non-hydrogen substituent on any available carbon or nitrogen atom, with as many optionally substituted groups as can be accommodated.

[0068] The term "polynucleotide" is used interchangeably with "nucleic acid" and refers to a polymer of nucleotides (ribonucleotides or deoxyribonucleotides). Polynucleotides formed from ribonucleotides can be referred to as "RNA polynucleotides", "ribonucleic acids" or simply "RNA"; polynucleotides formed from deoxyribonucleotides can be referred to as "DNA polynucleotides", "deoxyribonucleic acids" or simply "DNA". Polynucleotides can be single-stranded or double-stranded and optionally incorporate synthetic, non-natural or modified nucleotides capable of incorporation into DNA or RNA. An "artificial polynucleotide" refers to a polynucleotide whose sequence does not exist in nature or has been altered by human intervention.

[0069] As used herein, the term "messenger RNA" (abbreviated "mRNA") refers to any RNA polynucleotide encoding a target polypeptide that is capable of being 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.

[0070] As used herein, "open reading frame" or "ORF" refers to a sequence of several nucleotide triplets that encodes a polypeptide, i.e., can be translated into a polypeptide sequence.

[0071] As used herein, a "DNA construct" refers to an artificial polynucleotide comprising a sequence of interest operably linked to an expression promoter that controls the expression of the sequence of interest.

[0072] As used herein, an "expression vector" refers to a vector used to introduce a specific nucleic acid (usually 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.

[0073] The term "polypeptide" refers to any peptide or protein comprising two or more amino acids linked to each other by peptide bonds or modified peptide bonds (i.e., peptide isosteres). "Polypeptide" refers both to short chains, commonly referred to as peptides, oligopeptides or oligomers, and to longer chains commonly referred to as proteins.

[0074] As used herein, the expression "therapeutically effective amount" refers to an amount of a compound that, when administered, is sufficient to prevent one or more symptoms of the disease against which it is targeted or to alleviate the symptoms to some extent. The specific dose of the compound administered according to the present invention will of course be determined by the specific circumstances surrounding the case, including the compound being administered, the route of administration, the specific disorder being treated, the specific circumstances of the individual subject being treated and similar considerations. The term "drug" also encompasses the concept of "veterinary composition". Thus, they relate to compositions that are therapeutically effective when administered by any desired or applicable route to any animal (including humans).

[0075] According to the present invention, the term "antigen" is a compound that can be recognized by the immunoglobulin receptor of a B cell or, when complexed with MHC, by the T cell receptor. Preferably, the "antigen" is a polypeptide.

[0076] As used herein, the term "nanoparticle" refers to a particle having at least two dimensions in the nanoscale, particularly all three dimensions in the nanoscale, where the nanoscale ranges from about 1 nm to about 500 nm. Specifically, when the nanoparticle is substantially rod-shaped with a substantially circular cross-section, such as a nanowire or nanotube, the "nanoparticle" refers to a particle having at least two dimensions in the nanoscale, and these two dimensions are the cross-section of the nanoparticle. In the context of a nanoparticle composition, the "size" or "average size" as used herein refers to the average diameter of the nanoparticle composition. The term "lipid nanoparticle" as used herein refers to a nanoparticle whose outer shell is made entirely or partially of lipid.

[0077] As used herein, the "polydispersity index (PDI)" is a ratio that describes the uniformity of the particle size distribution of a system. A smaller value, such as less than 0.3, indicates a narrow particle size distribution.

[0078] As used herein, the term "ζ potential" is, for example, the electrokinetic potential of the lipid in a particle composition. The ζ potential is also defined as the potential difference between the dispersion medium and the stationary layer of fluid attached to the dispersed particles. It is generally considered a quantification of the charge magnitude and is often the only available way to characterize the properties of the double layer.

[0079] As used herein, the "apparent pKa" refers to an experimentally determined value derived from the average ratio of all ionized groups to deionized groups in a nanoparticle. The apparent pKa is different from the inherent pKa of any single molecule and is an important parameter for the performance of nanoparticles encapsulating RNA. The apparent pKa of nanoparticles can be measured by different techniques known in the art. For example, acid-base titration and 2-(p-toluidino)-6-naphthalenesulfonic acid (TNS) fluorescence methods are widely used in the art. Nanoparticles with an optimal pKa carry negligible charge at physiological pH, which prevents non-specific binding and in vivo toxicity. The optimal pKa of nanoparticles plays an important role in the endosomal escape mechanism and the release of RNA into the cytosol to exert a therapeutic effect.

[0080] Ionizable lipid

[0081] As described above, a first aspect of the present invention relates to an ionizable lipid of formula (I):

[0082]

[0083] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A, B, m, n, p, q, t, R3, X and Z are as defined above.

[0084] In one embodiment of the ionizable lipid, when n = t = m = 0, q = p = 1,

[0085] X is

[0086] wherein one of j and j' is 0 and the other is 1,

[0087] when D is -O-C(O)-, A = -C(O)-NH-, B = -NH-C(O)-, Z is R1, and R1 is a C1-C18 alkyl group,

[0088] R2 and R'2 are not C7-C18 alkyl groups.

[0089] In one embodiment, the ionizable lipid is a compound of formula (II)

[0090]

[0091] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A, B, D n, m, p, q, t, q + t, R1 and R2 are as defined above,

[0092] provided that

[0093] - when q + t = 1, X = -D-R2, D = -O-C(O)-, A = -NH-C(O)-, B = -NH-C(O)-, Z = R1, and R1 is a straight-chain or branched alkyl group, the moiety -(CH2) m - R1 contains at least 7 carbon atoms;

[0094] - excluding the compounds S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-homocysteine methyl ester,

[0095]

[0096] S-(2-methoxy-2-oxoethyl)-N-pyridin-2-ylcarbonyl)-L-cysteine methyl ester

[0097]

[0098] and (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole.

[0099] In another embodiment of the ionizable lipids 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):

[0100]

[0101] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0102] A is

[0103] B is

[0104] D is

[0105] n, m, and p are independently 0, 1, 2, 3, 4, 5, or 6;

[0106] q is selected from 1, 2, and 3;

[0107] t is selected from 0, 1, and 2;

[0108] and wherein q + t = 1, 2, or 3;

[0109] R1 and R2 are independently straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, wherein R1 and R2 are optionally substituted with one or more substituents selected from -OH, -COOR4, and -C(=O)S-R4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl;

[0110] and wherein

[0111] R3 is a heterocycle containing at least one N atom, or R3 is:

[0112]

[0113] wherein Ra and Rb are independently straight-chain or branched C1-C6 alkyl optionally substituted with a hydroxyl group;

[0114] provided that

[0115] - when q + t = 1, X = -D-R2, D = -O-C(O)-, A = -NH-C(O)-, B = -NH-C(O)-, Z = R1, and R1 is straight-chain or branched alkyl, the moiety -(CH2) m - R1 contains at least 7 carbon atoms; and

[0116] - excluding the compound S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-homocysteine methyl ester,

[0117]

[0118] S-(2-Methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-L-cysteine methyl ester

[0119]

[0120] and (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole.

[0121] As used herein, the term "pharmaceutically acceptable salt" includes any salt formed from a pharmaceutically acceptable non-toxic acid (including inorganic or organic acids). There are no restrictions on the salts other than that they must be pharmaceutically acceptable if used for therapeutic purposes. The preparation of pharmaceutically acceptable salts of the ionizable lipids of the present disclosure can be carried out by methods known in the art. For example, they can be prepared by conventional chemical methods from parent compounds containing basic or acidic moieties. Generally, such salts are prepared, for example, by reacting the free acid or base form of the ionizable lipids of the present disclosure with a stoichiometric amount of the suitable pharmaceutically acceptable base or acid in water or an organic solvent or in a mixture thereof.

[0122] 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, camphorsulfonic acid, mandelic acid, benzenesulfonic acid, p-toluenesulfonic acid, oxalic acid, methanesulfonic acid or naphthalenesulfonic acid; and base addition salts formed with alkali metals and alkaline earth metals and organic bases such as N,N-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, glucosamine, lysine and procaine. The ionizable lipids of the present disclosure and their salts may differ in certain physical properties, but they are equivalent for the purposes of the present invention.

[0123] The ionizable lipids of the present disclosure have asymmetric centers and can thus give rise to various stereoisomers. As used herein, the term "stereoisomers" refers to all isomers of a single ionizable lipid that differ only in the orientation of their atoms in space. The term stereoisomers includes enantiomers, racemates, racemic mixtures, geometric isomers (cis / trans or syn / anti or E / Z) and diastereoisomers. The present invention relates to each of these stereoisomers and mixtures thereof.

[0124] The preparation methods described herein can be modified to give enantiomerically pure compounds as well as mixtures of stereoisomers. Specific stereoisomers or specific mixtures can be prepared by a variety of methods, including using stereospecific reagents or by introducing chiral centers into the compounds during the preparation process. In addition, once the compounds are prepared, the stereoisomers can be separated by standard resolution techniques known to those skilled in the art.

[0125] In all embodiments of the invention involving the ionizable lipids of the present disclosure, even if not specifically mentioned, pharmaceutically acceptable salts or stereoisomers of the ionizable lipids or stereoisomers of their pharmaceutically acceptable salts are always contemplated.

[0126] The ionizable lipids of the present disclosure also include isotopes of the depicted structures. "Isotopes" refer to atoms having the same atomic number but different mass numbers due to different numbers of neutrons in the atomic nucleus. For example, isotopes include but are not limited to tritium, deuterium, 13 C or 14 C or 15 N. In addition, the compounds or salts of the present disclosure can be prepared by combining with solvents or water molecules by conventional methods to form solvates and hydrates.

[0127] The ionizable lipids of the present disclosure are characterized by their retention times, mass spectra, size distributions, polydispersity indices, and ζ potentials. These parameters can be measured by methods well known in the art. Some of these are described in more detail in the examples below.

[0128] 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; m is selected from 0, 1, 2, and 3.

[0129] Another embodiment relates to an ionizable lipid of formula (II):

[0130]

[0131] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0132] A is

[0133] B is

[0134] D is

[0135] n, m, and p are independently 0, 1, 2, 3, 4, 5, or 6;

[0136] q is selected from 1 and 2,

[0137] t is selected from 0 and 1,

[0138] and wherein q + t = 2;

[0139] R1 and R2 are independently a straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, wherein R1 and R2 are optionally substituted with one or more substituents selected from -OH, -COOR4 and -C(=O)S-R4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl,

[0140] and wherein

[0141] R3 is a heterocycle containing at least one N atom, or R3 is:

[0142]

[0143] wherein Ra and Rb are independently a straight-chain or branched C1-C6 alkyl optionally substituted with a hydroxyl group.

[0144] According to another embodiment, optionally in combination with one or more features of each of the above embodiments, R3 is a 5-membered or 6-membered ring containing one N atom and optionally a second heteroatom preferably selected from N and O.

[0145] In another embodiment, optionally in combination with one or more features of each of the above embodiments, R3 is selected from the following structures:

[0146]

[0147] In another embodiment, optionally in combination with one or more features of each of the above embodiments, 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.

[0148] In another embodiment, n is 2 or 3, m = t = 0, p = q = 2,

[0149]

[0150] wherein

[0151] R1 is a C10-C20 branched alkyl;

[0152] R2 is a C2-C30 straight-chain or branched alkyl or a C6-C24 straight-chain alkenyl or alkynyl; and

[0153] R3 is

[0154] wherein Ra = Rb = C1-C4 alkyl.

[0155] In another embodiment, n = 2, R2 is a C2-C30 straight-chain or branched alkyl, and wherein R3 is:

[0156]

[0157] wherein Ra = Rb = an alcohol-terminated C1-C4 alkyl.

[0158] In another embodiment, n is 2 or 3, m = t = 0, p = q = 2,

[0159] A is

[0160] R1 is a C10-C20 branched alkyl, R2 is a C3-C30 straight-chain or branched alkyl, and wherein R3 is an imidazole ring.

[0161] In another embodiment, n = 3, m = t = 0, p = q = 2,

[0162] A is

[0163] R1 is a C10-C20 branched alkyl, R2 is a C4-C20 straight-chain or branched alkyl, and R3 is a pyrrolidine ring.

[0164] In another embodiment, n = 2, m = t = 0, p = q = 2,

[0165] A is

[0166] R1 is a C10-C20 straight-chain or branched alkyl, R2 is a C4-C20 straight-chain or branched alkyl, and wherein R3 is N-4-methylpiperazine.

[0167] In another embodiment, n is selected from 2 or 3,

[0168] m = t = 0, and p = q = 2,

[0169]

[0170] R1 is a C10-C22 straight-chain or branched alkyl, alkenyl or alkynyl; R2 is a C3-C30 straight-chain or branched alkyl or a C3-C30 straight-chain alkenyl or alkynyl; and wherein R3 is a morpholine ring.

[0171] In another embodiment, n is selected from 2 or 3, m = 0, p = 2,

[0172] q = t = 1,

[0173] A is

[0174] B = D, R1 is a C10-C22 straight-chain or branched alkyl group; R2 is a C3-C30 straight-chain or branched alkyl group, a C3-C30 straight-chain alkenyl group or an alkynyl group, and R3 is a morpholine ring.

[0175] In another embodiment, n = q = p = 2; t = m = 0;

[0176] A is B is D is

[0177] R1 is a C10-C22 branched alkyl group;

[0178] R2 is a C8-C20 straight-chain alkenyl group or an alkynyl group; or a C3-C28 straight-chain alkyl group optionally substituted with one or more substituents selected from OH, -COOR4 and -COSR4, where R4 is a C1-C6 alkyl group, a C2-C6 alkenyl group or a C2-C6 alkynyl group; or a C12-C25 branched alkyl group;

[0179] and R3 is -N(CH3)2 or

[0180] In another embodiment, the ionizable lipid of the present disclosure is a compound of formula (III)

[0181]

[0182] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, where

[0183] A is -NH-C(O)- or -C(O)-NH-,

[0184] R1, R2 and R'2 are independently selected from straight-chain or branched C1-C30 alkyl groups, C2-C30 alkenyl groups, C2-C30 alkynyl groups, where each of R1, R2, R'2 is optionally substituted with one or more substituents selected from -OH, -COO(R4) and -COSR4, where R4 is a C1-C6 alkyl group, a C2-C6 alkenyl group or a C2-C6 alkynyl group; and

[0185] where B, D, n, m, p, q, t, q + t, j, j' and R3 are as defined above.

[0186] In a specific embodiment of the compound of formula (III), when n = t = m = 0, q = p = 1, A = B = -NH-C(O)-, where one of j and j' is 0 and the other is 1, D = -O-C(O)-, and where R1 is a C1-C18 alkyl group, R2 and R'2 are not C7-C18 alkyl groups.

[0187] In another embodiment, the ionizable lipid of the present disclosure is a compound of formula (IV)

[0188]

[0189] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0190] A is -NH-C(O)- or -C(O)-NH-

[0191] wherein B, D, n, m, p, q, t, q + t, f, f’, g, g’ and R3 are as defined above.

[0192] Examples of the ionizable lipids of formula (I), formula (II), formula (III) and formula (IV) include the compounds described in Table 1 below.

[0193]

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200]

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275] In one embodiment, the ionizable lipids of the present disclosure are compounds selected from the group consisting of tridecyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0099), 8-methylnonyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0101), tridecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0163), tetradecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0168), octadecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0169), hexadecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0170), tridecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0178), tetradecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0183), tridecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0194), 8-methylnonyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0196), dodecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0198), tetradecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0199), octadecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0200), as defined in the specification.Hexadecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0201), 6-Methylheptyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0202), Octyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0207), Tridecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0209), 8-Methylnonyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0211), Dodecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0213), Tetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0214), Octadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0215), Hexadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0216), Tridecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((3-(pyrrolidin-1-yl)propyl)amino)butyl)thio)propionate (VC-LC-0241), Tridecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0256), 8-Methylnonyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0258), 8-Methylnonyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0261), Octadecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0262)Hexadecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0263), Hexyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0268), Octyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0269), Tridecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-oleoylamido-4-oxobutyl)thio)propionate (VC-LC-0289), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butyrate (VC-LC-0294), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0296), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0297), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0298), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-(((3-oxo-3-(tetradecyloxy)propyl)thio)methyl)butyrate (VC-LC-0299), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0300), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0301), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0302)4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid nonadec-9-yl ester (VC-LC-0304), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(hexyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid nonadec-9-yl ester (VC-LC-0306), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid nonadec-9-yl ester (VC-LC-0307), 3-((4-((3-morpholinopropyl)amino)-3-oleoylamino-4-oxobutyl)thio)propanoic acid 2-decyltetradecyl ester (VC-LC-0353), 3-((4-((3-morpholinopropyl)amino)-3-oleoylamino-4-oxobutyl)thio)propanoic acid octadec-9-en-1-yl ester (VC-LC-0355), 3-((4-((1-methylpiperidin-4-yl)amino)-3-oleoylamino-4-oxobutyl)thio)propanoic acid 2-decyltetradecyl ester (VC-LC-0356), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0362), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-(dodecyloxy)-3-oxopropyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0366), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0367), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-(hexadecyloxy)-3-oxopropyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0369), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0370), 3-((4-((3-morpholinopropyl)amino)-4-oxo-3-stearoylaminobutyl)thio)propanoic acid 2-decyltetradecyl ester (VC-LC-0389), 6-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobut-2-yl)amino)-6-oxohexanoic acid 2-hexyldecyl ester (VC-LC-0418),2-decyltetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0428), dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0429), octadec-9-en-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0430), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0431), heptadec-9-yl 4-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0439), heptadec-9-yl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0440), heptadec-9-yl 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoate (VC-LC-0441), heptadec-9-yl 4-((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0442), (Z)-heptadec-9-yl 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)butanoate (VC-LC-0443), heptadec-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)propionate (VC-LC-0473), 2-hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0474), 8-methylnonyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0475), dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0477)Tetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0478), hexadecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0480), 2-decyltetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0487), dec-2-yn-1-yl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0488), octadec-9-en-1-yl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0489), tridecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0490), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0491), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0492), 2-decyltetradecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0504), dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0505), tridecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0507), 2-hexyldecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0508), 8-methylnonyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0509)2-Ethylhexyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0510), 6-Methylheptyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0515), 2-Decyltetradecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0521), 2-Hexyldecyl 4-((1-methylpiperidin-4-yl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butyrate (VC-LC-0524), 2-Hexyldecyl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyrate (VC-LC-0525), 2-Hexyldecyl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyrate (VC-LC-0531), (Z)-2-Hexyldecyl 4-((1-methylpiperidin-4-yl)amino)-2-(((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0539), Heptadec-9-yl 4-((3-morpholinopropyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butyrate (VC-LC-0540), Heptadec-9-yl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0541), Heptadec-9-yl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0542), Heptadec-9-yl 2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0543), Heptadec-9-yl 2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0544), Heptadec-9-yl 4-((3-morpholinopropyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0546),Heptadec-9-yl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0547), Heptadec-9-yl 2-(((3-((6-methylheptyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0548), Heptadec-9-yl 2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0549), Heptadec-9-yl 2-(((3-(hexyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0550), Heptadec-9-yl 4-((3-morpholinopropyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0551), Heptadec-9-yl 2-(((3-((2-decyltetradecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0553), Heptadec-9-yl 2-(((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0554), Tridecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0556), 2-Hexyldecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0557), 8-Methylnonyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0558), 2-Ethylhexyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0559), 2-Hexyldecyl 4-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0606), 2-Hexyldecyl 4-((3-((8-methylnonyl)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)butyrate (VC-LC-0608), 2-hexyldecyl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butyrate (VC-LC-0609), 2-hexyldecyl 2-((3-morpholinopropyl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butyrate (VC-LC-0610), 2-hexyldecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0639), 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0729), 8-methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0730), 2-ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0731), dodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0732), tetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0733), octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0734), hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0735), 6-methylheptyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0736), docosyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0737), hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0738)Octyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0739), 2-decyltetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0741), 2-dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0742), octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (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-triazatricos-21-oate (VC-LC-0757), tridecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0796), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0797), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0798), 2-ethylhexyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0799), dodecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0800), tetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0801), octadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0802), hexadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0803)6-Methylheptyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0804), Docosyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0805), Hexyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0806), Octyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0807), 2-Decyltetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0809), Dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0810), Octadec-9-en-1-yl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0811), Tridecyl 3-((4-(4-(2-hydroxyethyl)piperazin-1-yl)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0828), 8-Methylnonyl 3-((3-(2-octyldodecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0848), Tridecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0864), 2-Hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0865), 8-Methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0866), 2-Ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0867), Dodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0868)Tetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0869), Octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0870), Hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0871), 6-Methylheptyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0872), Docosyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0873), Hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0876), Octyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0877), Dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0880), Octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0881), 6-Methylheptyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0890), Octyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0895), 2-Hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0919), 2-Ethylhexyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0921), Dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0922)Tridecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0936), 2-Hexyldecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0937), Dodecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0940), 6-Methylheptyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0944), Docosyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0945), Octyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0949), (Z)-Octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0953), 2-Hexyldecyl 6-((4-((3-((2-hexyldecyloxy)-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)propionate (VC-LC-1065), Octyl 7-(2-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)ethyl)-2-methyl-14-((3-(octyloxy)-3-oxopropyl)thio)-6,9-dioxo-17-thia-2,5,8-triazadocos-20-oate (VC-LC-1068), Dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-1-oxo-4-((3-oxo-3-(tridecyloxy)propyl)thio)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyldiyl))dipropionate (VC-LC-1082)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-1083), Dioctyl 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), Dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1085), Dioctyl 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))dipropionate (VC-LC-1088), Dioctyl 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), 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), Dioctyl 3,3'-((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)Dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(deca-2-yn-1-yloxy)-3-oxopropyl)sulfanyl)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1095), Dioctyl 3,3’-((8-oxo-8-((1-oxo-4-((3-oxo-3-(tridecyloxy)propyl)sulfanyl)-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-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1097), Dioctyl 3,3’-((8-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1098), Dioctyl 3,3’-((8-((4-((3-((6-methylheptyl)oxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1105), Dioctyl (Z)-3,3’-((8-((4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1111), (9Z,12Z)-Octadeca-9,12-dien-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)sulfanyl)propionate (VC-LC-1143), (2E,6E)-3,7,11-Trimethyldodeca-2,6,10-trien-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)sulfanyl)propionate (VC-LC-1192), Bis((E)-3,7-dimethylocta-2,6-dien-1-yl) 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)sulfanyl)methyl)succinate (VC-LC-1194)Tetradecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-1202), dodecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-1203), bis(3,7-dimethyloctyl) 2-(((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)propionate (VC-LC-1224), (9Z,12Z)-octadeca-9,12-dien-1-yl 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-1230), 2-hexyldecyl 2-((2-(dimethylamino)ethyl)carbamoyl)-4-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)butanoate (VC-LC-1254), bis(3,7-dimethyloctyl) 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyloxy)carbonyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1281), bis((E)-3,7-dimethyloct-2,6-dien-1-yl) 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyloxy)carbonyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1282), bis((E)-3,7-dimethyloct-2,6-dien-1-yl) 2-(((4-((2-butyloctyloxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1291), bis(3,7-dimethyloctyl) 2-(((4-((2-decyltetradecyloxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1295), bis((E)-3,7-dimethyloct-2,6-dien-1-yl) 2-(((4-((2-decyltetradecyloxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1296), or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them.

[0276] In another embodiment, the ionizable lipid of formula (I) is a compound selected from: 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-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-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-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 stereoisomer of any of them.

[0277] In a particular embodiment, the ionizable lipid of formula (I) is a compound selected from: 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, as described herein.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 stereoisomer of any of them.

[0278] Preparation method

[0279] The method for preparing the ionizable lipid of formula (I) is also part of the present invention.

[0280] All reactants and solvents required for preparing the compounds of the present disclosure are commercially available.

[0281] For example, the ionizable lipid of formula (I) can be prepared according to any of the following synthetic schemes. Any person skilled in the art will know which reactants are required to obtain any specific ionizable lipid according to the present disclosure in accordance with the synthetic methods described in the following schemes or similar methods.

[0282] The ionizable lipid of formula (II) wherein B is an amide moiety and t = 0, q = 2, such as the compound of formula (IIa), can be prepared by the synthetic method depicted in Scheme 1 below or a similar method.

[0283]

[0284] Examples of the acid (R 1 -COOH), amine (AM-NH2), and acrylate (CH2=CH-(CO)-O-R2) that can be used to prepare the compound of formula (II) according to Scheme 1 are listed in Tables 2, 3, and 4 below, respectively.

[0285] Table 2

[0286]

[0287] Table 3

[0288]

[0289]

[0290] Table 4

[0291]

[0292] Among them, the ionizable lipid of formula (II) where B is an amide moiety and t = q = 1, such as the compound of formula (IIb) or its analogs, can be prepared by the synthetic methods depicted in Scheme 2 below.

[0293]

[0294]

[0295] The ionizable lipid of formula (I) where B is an ester moiety, such as the compound of formula (IIc) or its analogs or the compound of formula (IId) or its analogous compounds, can be prepared by the synthetic methods depicted in Scheme 3 or 4 below or analogous methods.

[0296] Lipid analogs having amide - ester substitution, namely the compounds of formula (IIc) (where B = C, t = 0, q = 2) and formula (IId) (where B = C, t = q = 1), can be prepared according to Synthetic Schemes 3 and 4 below:

[0297]

[0298] Depending on whether the reaction starts from 2 - oxotetrahydrothiophene - 3 - carboxylic acid (Compound A) or 5 - oxotetrahydrothiophene - 3 - carboxylic acid (Compound B), two different families of compounds (i.e., the compounds of formula (IIc) or the compounds of formula (IId) respectively) can be obtained. 2 - Oxotetrahydrothiophene - 3 - carboxylic acid and 5 - oxotetrahydrothiophene - 3 - carboxylic acid can be prepared as described in the literature (refer to Garbiras, B.J., Marburg, S., "Preparation of Carboxythiolactones and Their Active Derivatives", Synthesis, 1999, vol. 2, pp. 270 - 274).

[0299] Possible alcohols R 4 -OH used for preparing the compounds of formula (IIc) and formula (IId) according to Schemes 3 and 4 are listed in Table 5 below. The amine AM - NH2 and the acrylate CH2 = CH(CO)OR2 are those listed in Tables 3 and 4 above.

[0300] Table 5

[0301]

[0302] The ionizable lipid of formula (I) where B is an amide moiety, such as the compound of formula (IIe) (A = B = amide, t = 2, q = 1) or its analogs, can be prepared by the synthetic methods depicted in Scheme 5 below or analogous methods.

[0303]

[0304] Wherein B is an ionizable lipid of formula (I) of the amide moiety, such as a compound of formula (IIf) (A = B = amide, t = 0, q = 3) or an analogue thereof, and can be prepared by the synthetic method depicted in Scheme 6 below or a similar method.

[0305]

[0306] Compounds C and D are commercially available.

[0307] Wherein B is an ionizable lipid of formula (I) of the amide moiety (A = B = amide), t = 0 and q = 1, or an analogue thereof, and can be prepared as disclosed in FR3104943A1.

[0308] The ionizable lipid of formula (I), such as wherein Z is R1,

[0309] X is

[0310] a compound of formula (III), such as a compound of formula (IIIa) wherein A = B = amide, t = 0, q = 3, or an analogue thereof, and can be prepared by the synthetic method depicted in Scheme 7 below or a similar method.

[0311]

[0312] The ionizable lipid of formula (I), such as wherein X is -D-R2 and

[0313] Z is

[0314]

[0315] a compound of formula (IV), such as a compound of formula (IVa) wherein A = B = -NH-C(O)-, t = 0, q = 2, or an analogue thereof, and can be prepared by the synthetic method depicted in Scheme 8 below or a similar method.

[0316]

[0317] Lipid nanoparticle

[0318] As described above, the ionizable lipids of the present disclosure can form LNPs in solution.

[0319] Accordingly, the present invention also relates to an LNP comprising an ionizable lipid of formula (I) as defined herein (in particular a lipid of formula (II), (III) or (IV)), including ionizable lipids not claimed above and below.

[0320] All embodiments described for the ionizable lipid of formula (I) are also applicable to the LNP.

[0321] Generally, the LNP has a core-shell structure comprising a core and a shell. LNP formulations with lipid components having several different properties such as ionizable lipids, sterols, polyethylene glycolylated lipids, and non-cationic lipids (also referred to as "helper lipids") such as phospholipids contain several phases, which phase-separate into a hydrophobic core region formed by the ionizable lipid and cholesterol, and a surrounding shell covering the surface formed by the helper lipid, cholesterol, and polyethylene glycolylated lipid. More specifically, the ionizable lipid of formula (I) forms part of the inner shell of the LNP, and optionally, the pharmaceutically active agent is encapsulated or loaded in the core.

[0322] In one embodiment, optionally in combination with one or more features of the above-described individual embodiments, the LNP further comprises at least one lipid selected from non-cationic lipids, sterols or steroid precursors, and PEG-modified lipids.

[0323] In another embodiment, optionally in combination with one or more features of each of the above embodiments, the LNP comprises an ionizable lipid of the present disclosure and a non-cationic lipid. Examples of non-cationic lipids 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 (SOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-docosanoyl-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-cholesteryl succinyl-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 16.0PE), 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. In one embodiment, the non-cationic lipid is DSPC. In a particular embodiment, the non-cationic lipid is DOPE. In another particular embodiment, the lipid-containing particle comprises DSPC and DOPE.

[0324] In another embodiment, optionally in combination with one or more features of each of the above embodiments, the LNP further comprises a sterol or sterol precursor.

[0325] In another embodiment, optionally in combination with one or more features of each of the above embodiments, the LNP comprises an ionizable lipid of the present disclosure and a sterol or sterol precursor.

[0326] Examples of sterols include, but are not limited to, cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof. In one particular embodiment, optionally in combination with one or more features of the various embodiments above, the sterol is cholesterol. Examples of sterol precursors include, but are not limited to, triterpenes, triterpenoids, or such steroid precursors. Non-limiting examples of triterpenes, triterpenoids, and other steroid precursors include squalene, achilleol, polypodatetrane, lanostane, cucurbitacin, hopane, oleanane, chamaecydin, lupine, and mixtures thereof.

[0327] In another embodiment, optionally in combination with one or more features of the various embodiments 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.

[0328] In another embodiment, optionally in combination with one or more features of the various embodiments above, the LNP further comprises a PEGylated lipid. The term "PEGylated lipid" refers to a lipid comprising a polyethylene moiety. Examples of PEGylated lipids include, but are not limited to, PEGylated phosphatidylethanolamine, PEGylated phosphatidic acid, PEGylated phosphatidylcholine, PEGylated ceramide, PEGylated dialkylamine, PEGylated diacylglycerol, PEGylated dialkylglycerol, and mixtures thereof. In some embodiments, the PEGylated lipid is PEG-DMG, PEG-c-DOMG (also known as PEG-DOMG), PEG-DSG, PEG-DPG, or a combination thereof.

[0329] In another embodiment, optionally in combination with one or more features of the various embodiments above, the LNP further comprises a conjugated lipid.

[0330] Examples of conjugated lipids include, but are not limited to, poly(sarcosine) (pSar) lipids and derivatives, such as N-tetradecyl-pSar25, N-hexadecyl-pSar25, N-octadecyl-pSar25, N-dodecyl-pSar25, DMG-pSar25, 18:1PE(DOPE)pSar25, N-tetraamine-pSar25, N-tetraamine-pSar35, N-tetraamine-pSar45, N-tetraamine-pSar45-maleimide.

[0331] In another embodiment, optionally in combination with one or more features of each of the above embodiments, 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.

[0332] In one embodiment, optionally in combination with one or more features of each of the above embodiments, the lipid component of the LNP comprises, or consists of, an ionizable lipid, a non-cationic lipid, a sterol, and a PEG-modified lipid as defined herein.

[0333] In a specific embodiment, optionally in combination with one or more features of each of the above embodiments, the lipid component of the LNP comprises an ionizable lipid disclosed herein and at least one of distearoyl phosphatidylcholine (DSPC), cholesterol, and DMG-PEG2000. In a more specific embodiment, the lipid component comprises, or consists of, an ionizable lipid disclosed herein, DSPC, cholesterol, and DMG-PEG2000.

[0334] In another embodiment, optionally in combination with one or more features of each of the above embodiments, the LNP comprises 25 to 60 mole percent (mol%) of an ionizable lipid, 0.1 to 10 mol% of a PEG-modified lipid or conjugated lipid (particularly a PEG-modified lipid), 10 to 45 mol% of a non-cationic lipid, and 10 to 40 mol% of a sterol. In particular, the LNP comprises 32 to 50 mol% of an ionizable lipid, 1 to 8 mol% of a PEG-modified lipid or conjugated lipid (particularly a PEG-modified lipid), 12.5 to 42 mol% of a non-cationic lipid, and 15 to 38.5 mol% of a sterol. More particularly, the LNP comprises 35 to 47 mol% of an ionizable lipid, 1 to 4 mol% of a PEG-modified lipid, 30 to 38 mol% of a non-cationic lipid, and 15 to 25 mol% of a sterol.

[0335] In another embodiment of the LNP, optionally in combination with one or more features of each of the above embodiments, the amount of the ionizable lipid is 25 to 64 mol%, the amount of the PEG-modified lipid is 0.1 to 1.5 mol%, and the amount of the sterol is 35 to 74 mol%.

[0336] As used herein, "mol%" refers to the mole percentage of a component relative to the total number of moles of all lipid components in the LNP (i.e., the total number of moles of ionizable lipid, PEG-modified lipid, non-cationic lipid, and sterol).

[0337] In another embodiment, optionally in combination with one or more features of each of the above embodiments, the molar ratio of ionizable lipid to non-cationic lipid is in the range of 6:1 to 1:2 or 2:1 to 1:1.

[0338] In another embodiment, optionally in combination with one or more features of each of the above embodiments, the molar ratio of ionizable lipid to sterol is in the range of 5:1 to 1:2 or 2:1 to 1:1.

[0339] In another embodiment, optionally in combination with one or more features of each of the above embodiments, the molar ratio of ionizable lipid to PEGylated lipid is in the range of 120:1 to 2:1 or 100:1 to 10:1.

[0340] Active agent

[0341] As described above, the LNPs of the present invention may comprise one or more pharmaceutically active agents.

[0342] As used herein, the term "pharmaceutically active agent" refers to an agent having pharmacological activity and used for curing, alleviating, treating or preventing diseases in a subject, particularly a human.

[0343] For the purposes of the present invention, pharmaceutically active agents include low molecular weight drugs, polynucleotides, peptides, antibodies, proteins and combinations thereof.

[0344] As used herein, the term "polynucleotide" refers to natural or artificial deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). The polynucleotide may comprise at least one chemical modification 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-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine (also known as m5U), 5-methoxyuridine, 2'-O-methyluridine and combinations thereof. In particular, the chemical modification is N1-methylpseudouridine, 5-methoxyuridine or a combination thereof; in particular, the chemical modification is N1-methylpseudouridine.

[0345] The polynucleotide is modified with 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% of N1-methylpseudouridine moieties, or is fully modified with N1-methylpseudouridine, 5-methoxyuridine or a combination thereof.

[0346] In a particular embodiment, optionally in combination with any of the embodiments provided above, the active agent is selected from polynucleotides, DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide, and expression vectors containing a DNA construct comprising a promoter operably linked to a sequence encoding a polynucleotide.

[0347] In a particular embodiment, optionally in combination with any of the embodiments provided above, the polynucleotide is ribonucleic acid (RNA).

[0348] Specifically, the RNA is selected from 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.

[0349] In a particular embodiment, optionally in combination with any of the embodiments provided above, the RNA is mRNA.

[0350] Those skilled in the art know how to produce the polynucleotide, DNA construct or expression vector by conventional methods well known in the art (e.g., by chemical synthesis or by molecular biology techniques) without exercising any inventive skills.

[0351] In a particular embodiment, optionally in combination with any of the embodiments provided above, the ratio of ionizable lipid to RNA in the LNP (N / P; where N represents the number of moles of amine present in the ionizable lipid and P represents the number of moles of phosphate ester present in the polynucleotide backbone) is in the range of 20:1 to 2:1, particularly 10:1 to 3:1.

[0352] In a particular embodiment, optionally in combination with any of the embodiments provided above, the polynucleotide is an isolated artificial polynucleotide.

[0353] In one embodiment, optionally in combination with one or more features of the above-described embodiments, the LNP comprises an ionizable lipid, a non-cationic lipid, a sterol, a PEG-modified lipid, and a polynucleotide as defined herein.

[0354] The LNP containing one or more polynucleotides can be prepared by standard methods, such as microfluidic mixing as disclosed in Hassett, K. J. et al. ("Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines", 2019, Mol. Ther. Nucleic Acid, vol. 15, pp. 1-11), or manual / batch 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 those skilled in the art will know how to proceed in each specific case.

[0355] Generally, the method for preparing LNP includes: i) preparing a first alcoholic mixture in a suitable alcohol such as ethanol, which contains the ionizable lipid of the present disclosure and optionally at least one lipid selected from non-cationic lipids, sterols, and PEG-modified lipids; ii) preparing a second aqueous composition, which contains polynucleotides and an acidified buffer; and iii) mixing i) and ii) in a microfluidic mixer. The microfluidic mixer allows for thorough and rapid mixing of the lipid phase and the polynucleotide phase in a microdevice. Depending on the process parameters, especially the total flow rate, those skilled in the art will be able to adjust the size of the LNP.

[0356] In a specific embodiment, optionally in combination with any of the embodiments provided above, the polynucleotide encodes a polypeptide, specifically, wherein the polypeptide is an antigen. More specifically, the antigen is selected from viral proteins, bacterial proteins, and tumor-associated antigens.

[0357] In a specific embodiment, optionally in combination with any of the embodiments provided above, the polypeptide is an antibody or a fragment thereof. In a more specific embodiment, the antibody or a fragment thereof is a therapeutic antibody or a fragment thereof.

[0358] In another embodiment, optionally in combination with any of the embodiments provided above, the antigen is a SARS-CoV-2 antigen, particularly a SARS-CoV-2 spike antigen.

[0359] Methods for the preparation of the above-described lipids, LNPs, and pharmaceutical compositions are described herein and / or are known in the art. Depending on the intended use of the composition, one of ordinary skill in the art will know which LNP to use to encapsulate each pharmaceutical active agent. Specifically, methods for the synthesis of compositions formed from LNPs encapsulating RNA are well known to those of ordinary skill in the art and are well established in molecular biology protocols. Specific conditions are indicated in the examples.

[0360] As described above, another aspect of the invention relates to a pharmaceutical composition comprising an LNP as defined herein and a pharmaceutically acceptable excipient or carrier.

[0361] The expression "pharmaceutically acceptable excipient or carrier" refers to a pharmaceutically acceptable material, composition, or carrier. Each component must be pharmaceutically acceptable in the sense of being compatible with the other ingredients 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 producing excessive toxicity, irritation, allergic responses, immunogenicity, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0362] Examples of suitable pharmaceutically acceptable excipients are solvents, dispersion media, diluents, or other liquid carriers, dispersing or suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc. The use of any conventional excipient medium is considered to be within the scope of the invention unless it is incompatible with the substance or its derivatives, such as producing any undesirable biological effects or interacting in a harmful manner with any other component of the pharmaceutical composition.

[0363] The relative amounts of the pharmaceutical active ingredient, pharmaceutically acceptable excipient, and / or any other ingredients in the pharmaceutical compositions of the invention vary depending on the identity, size, and / or condition of the subject being treated and further depending on the route of administration of the composition.

[0364] Pharmaceutically acceptable excipients for the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing agents, and / or granulating agents, surfactants and / or emulsifying agents, disintegrants, binders, preservatives, buffering agents, lubricants, and / or oils. Excipients such as coloring agents, coating agents, sweetening agents, and flavoring agents may be present in the composition, at the discretion of the formulator.

[0365] In one embodiment, optionally in combination with any of the embodiments provided above, the pharmaceutical compositions disclosed herein are administered orally, intranasally, intravenously, intraperitoneally, intramuscularly, intradermally, subcutaneously, topically or intra-articularly.

[0366] The pharmaceutical compositions of the present disclosure can be prepared by methods known in the art of pharmacy. For example, a pharmaceutical composition 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 LNP containing the pharmaceutically active agent or the pharmaceutical composition of the present disclosure can be used for therapeutic applications. Specifically, they can be used as non-viral vectors for general biopharmaceutical applications such as vaccines or gene therapy to effectively transfect genetic material into eukaryotic cells.

[0367] Accordingly, one aspect of the present invention relates to an LNP or a pharmaceutical composition as defined herein for use in a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the nanoparticle composition or pharmaceutical composition as defined herein.

[0368] Another aspect of the present invention relates to an LNP or a pharmaceutical composition as defined herein for use in a method of inducing an immune response in a subject, for use in a method of therapeutic immunization of a subject, for use as a vaccine, or for use in gene therapy. In particular, the subject is a human.

[0369] In one embodiment, optionally in combination with any of the embodiments provided above, the disease or disorder is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular and renal vascular diseases, and metabolic diseases.

[0370] When the pharmaceutically active agent is a polynucleotide, the LNP or pharmaceutical composition described herein can be used for vaccine therapy, for enhancing the efficacy of conventional vaccines and / or as a novel vaccine form for combating infectious pathogens such as viruses, bacteria, fungi, protozoa, prions, and worms; or for treating diseases such as cancer and proliferative diseases.

[0371] In one embodiment, optionally in combination with any of the embodiments provided above, the pharmaceutical composition is a vaccine. In a more specific embodiment, the pharmaceutical composition is a vaccine and further comprises an adjuvant. Those skilled in the art will know, based on their common general knowledge, which excipients, carriers, and adjuvants should be included in the vaccine depending on the intended use.

[0372] Throughout this specification and the claims, the word "comprising" and its variations are not intended to exclude other technical features, additives, components or steps. Additionally, the word "comprising" encompasses the case of "consisting of".

[0373] The following examples and figures are provided by way of illustration and are not intended to limit the invention. Additionally, the invention encompasses all possible combinations of the specific and preferred embodiments described herein.

[0374] Examples

[0375] Reagents were purchased from Sigma - Aldrich, TCI Chemicals, Fluorochem or VWR. All possible sensible combinations shown previously were prepared according to a three - step reaction scheme.

[0376] Example 1 - Synthesis of Lipid VC - LC - 0163

[0377]

[0378] Step 1.

[0379] DL - homocysteine hydrochloride (776 mg, 5.0 mmol) was dissolved in 13 mL of anhydrous dichloromethane at room temperature. Then triethylamine (588 μL, 4.3 mmol) was 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 its completion was checked by TLC chromatography. The resulting product was purified by flash chromatography using a hexane / ethyl acetate gradient (100 / 0 to 0 / 100). The fractions containing the product were combined and evaporated under reduced pressure to give a pale yellow oil (1.39 g, 89%).

[0380] Step 2.

[0381] 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-morpholinopropanamine (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, and then the solvent was evaporated under reduced pressure. The dry mixture was redissolved in ethyl acetate (20 mL) and washed with 0.1 M HCl(aq) solution (20 mL, 3 consecutive washes), then with distilled water (20 mL, 3 consecutive washes), and finally with saturated brine (20 mL, 3 consecutive washes; NaCl: water prepared by adding 35 g of NaCl to 100 mL of water). The combined organic fractions were dehydrated with MgSO4 (anhydrous), and then evaporated under reduced pressure to give a pale yellow oil (351 mg, quantitative yield).

[0382] Step 3.

[0383] 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), and then tridecyl acrylate (29 μL, 0.1 mmol). The reaction mixture was stirred at room temperature overnight, and then its completion was checked by TLC chromatography. The product obtained was purified by column chromatography using a gradient of dichloromethane / dichloromethane:methanol:ammonium hydroxide (80:20:1) (100 / 0 to 0 / 100). The fractions containing the product were combined and evaporated under reduced pressure to give VC-LC-0163 as a pale yellow oil (64 mg, 78%).

[0384] VC-LC-0163 was characterized by HPLC-LSD-MS (HPLC was Waters Alliance ELSD 2424, column was Xbridge BEH C8, 4.6 mm, 50 mm, 2.5 μm, gradient was 5% to 95% B (A: water containing 0.01% TFA; B: acetonitrile containing 0.01% TFA), for 20 min, rate = 1 mL / min).

[0385] The injection volume was 2 μL and the column temperature was 65 °C.

[0386] Successful detection of the analyte was carried out in the positive mode using an evaporative light scattering detector and a simple quadrupole mass detector with a spectrophotometer Waters Acquity QDa.

[0387] Retention time: 15.76 minutes, MS(ES): experimental m / z [M + H]+ 754.82 {theoretical m / z [M + H]+ 754.61}.

[0388] Example 2 - Synthesis of Lipid VC-LC-0605

[0389]

[0390] Step 1.

[0391] Dissolve 2-oxotetrahydrothiophene-3-carboxylic acid (730 mg, 5 mmol) in 13 mL of anhydrous dichloromethane at room temperature. Then add N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (870 mg, 4.51 mmol), then add dimethylaminopyridine ((104 mg, 0.85 mmol) and 2-hexyl-1-decanol (1.03 g, 4.25 mmol). Stir the reaction overnight at room temperature and then check its completion by TLC chromatography. Purify the resulting product by flash chromatography using a gradient of hexane / ethyl acetate (100 / 0 to 0 / 100). Combine the fractions containing the product and evaporate under reduced pressure to give a pale yellow oil (1.33 g, 84%).

[0392] To obtain VC-LC-0605, perform Steps 2 and 3 in Example 1 (41% yield).

[0393] VC-LC-0605 was characterized by HPLC-LSD-MS (HPLC was WaterAlliance ELSD 2424, column was Xbridge BEH C8, 4.6 mm, 50 mm, 2.5 μm, gradient was 5% to 95% B (A: water containing 0.01% TFA; B: acetonitrile containing 0.01% TFA), for 20 min, rate = 1 mL / min.

[0394] The injection volume was 2 μL and the column temperature was 65 °C.

[0395] The analyte was successfully detected by an evaporative light scattering detector and a simple quadrupole mass detector in positive mode using a spectrophotometer WatersAcquity QDa.

[0396] Retention time: 16.99 minutes, MS(ES): experimental m / z (MH+) 769.61 {theoretical m / z [M+H]+ 769.79}

[0397] Example 3 - Synthesis of Lipid VC-LC-0540

[0398]

[0399] To obtain lipid VC-LC-0540, the procedure of Example 2 was followed, but starting from 5-oxotetrahydrothiophene-3-carboxylic acid instead of 2-oxotetrahydrothiophene-3-carboxylic acid (35% yield).

[0400] VC-LC-0540 was characterized by HPLC-LSD-MS (HPLC: WaterAlliance ELSD 2424, column: Xbridge BEH C8, 4.6 mm, 50 mm, 2.5 μm, gradient from 5% to 95% acetonitrile B (A: water containing 0.01% TFA; B: acetonitrile containing 0.01% TFA), lasting 20 min, rate = 1 mL / min.

[0401] The injection volume was 2 μL and the temperature in the column was 65 °C.

[0402] Successful detection of the analytes was performed using a spectrophotometer Waters Acquity QDa by an evaporative light scattering detector and a simple quadrupole mass detector in positive mode.

[0403] Retention time: 17.2 min, MS (ES): experimental m / z (MH+) 783.63 {theoretical m / z [M+H]+ 783,80}.

[0404] Example 4 - Synthesis of lipid VC-LC-0749

[0405]

[0406] 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 product was then washed twice with distilled water (2 x 40 mL), with a saturated carbonate solution (40 mL), and finally with saturated brine (40 mL). The organic layer was dehydrated with anhydrous MgSO4, filtered, and evaporated under reduced pressure. The resulting residue (intermediate VC-LC-9096) was used directly without any further purification and was characterized by HPLC-ELSD-MS. Retention time: min 6.5, MS (ES): experimental m / z [M+H] + =306.15.

[0407]

[0408] Step 1b: Dissolve VC-LC-9096 (639 mg, 2.1 mmol) in 5 mL of THF and 0.1 mL of H2O at room temperature. Then add tris(2-carboxyethyl)phosphine hydrochloride (611 mg, 2.1 mmol), followed by octyl acrylate (1120 μL, 5.2 mmol) and triethylamine (1464 μL, 10.5 mmol). Stir the reaction mixture overnight at room temperature and then perform HPLC-ELSD-MS. Then evaporate the crude reaction under reduced pressure and wash it twice with distilled water (2 x 20 mL) and finally with saturated brine (20 mL). Purify the resulting residue by flash chromatography (hexane / ethyl acetate gradient: 100 / 0 to 0 / 100) to obtain 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.

[0409]

[0410] Step 2: Under an argon atmosphere, dissolve the intermediate compound VC-LC-9097 (100 mg, 0.15 mmol) in 1.5 mL of anhydrous tetrahydrofuran at room temperature. After the solid has completely dissolved, add N,N-diethylethylenediamine (215 μL, 1.5 mmol) using a syringe. Stir the reaction mixture at room temperature for 30 minutes and then use HPLC-MS to detect its completion. Then remove the solvent under reduced pressure. Subsequently, redissolve the dry crude reaction in 80 mL of ethyl acetate and wash it three times with 0.1 M HCl solution (1 x 10 mL), water (1 x 10 mL), and finally saturated brine (10 mL). Dry the organic layer over anhydrous MgSO4, filter, and evaporate under reduced pressure to obtain the intermediate compound VC-LC-9127 as a colorless oil (quantitative yield). VC-LC-9127 was characterized by HPLC-ELSD-MS. Retention time: min 13.8, MS(ES): experimental m / z [M+H]+ = 792.66.

[0411]

[0412] Step 3: Dissolve the intermediate compound VC-LC-9127 (120 mg, 0.15 mmol) in anhydrous tetrahydrofuran (1.5 mL) under an argon atmosphere. Then, add tetradecyl acrylate (86.0 μL, 0.30 mmol) using a syringe. Then stir the solution at room temperature for 1.5 hours (monitor the reaction by HPLC-MS), and then evaporate the solvent under reduced pressure. Purify the resulting residue by column chromatography (gradient of dichloromethane / eluent A: 100 / 0 to 0 / 100, eluent A: dichloromethane / methanol / ammonium hydroxide 80:20:1) to obtain VC-LC-0749 as an oil (83% yield).

[0413]

[0414] VC-LC-0749 was characterized by HPLC-ELSD-MS. Retention time: 18.75 min, MS(ES): experimental value m / z [M+H]+ 1060.96 {theoretical value m / z [M+H]+ 1060.74}.

[0415] Example 5 - Preparation of Lipid VC-LC-1194

[0416]

[0417] Step 1: Dissolve DL-homocysteine thiolactone hydrochloride (776 mg, 5.0 mmol) in 13 mL of dichloromethane at room temperature. Then add triethylamine (588 μL, 4.3 mmol), 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). Stir the reaction mixture at room temperature overnight, and then check its completion by TLC chromatography. Purify the resulting product by flash chromatography using a gradient of hexane / ethyl acetate (100 / 0 to 0 / 100). Combine the fractions containing the product and evaporate under reduced pressure to obtain a light yellow oil (1.39 g, 89%).

[0418] The product was characterized by HPLC-ELSD-MS. Retention time: 14.25 min, MS(ES): experimental value m / z [M+H]+ 356.44 {theoretical value m / z [M+H]+ 356.26}.

[0419]

[0420] 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, and then the solvent was evaporated under reduced pressure. The dry mixture was redissolved in ethyl acetate (20 mL) and washed with 0.1 M HCl(aq.) solution (20 mL, 3 consecutive washes), then with distilled water (20 mL, 3 consecutive washes), and finally with saturated brine (20 mL, 3 consecutive washes; NaCl: water prepared by adding 35 g of NaCl to 100 mL of water). The combined organic fractions were dried over MgSO4 (anhydrous), then evaporated under reduced pressure to give a pale yellow oil (quantitative yield).

[0421] The product was characterized by HPLC-LSD-MS. Retention time: 10.5 min, MS(ES): 444.37 experimental value m / z [M+H]+: 444.55 {theoretical value m / z [M+H]+}.

[0422] Step 3: Acrylate synthesis. Methylenesuccinic 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. Then 3,7-dimethyl-2,6-heptadien-1-ol (845 μL, 4.7 mmol) and triethylamine (932 μL, 6.7 mmol) were added. The reaction mixture was stirred at room temperature overnight under an argon atmosphere. The crude reaction product was then washed twice with distilled water (2 x 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 (gradient of hexane / ethyl acetate: 100 / 0 to 0 / 100) to give an oil.

[0423] The product was characterized using thin layer chromatography (100% hexane).

[0424]

[0425] Step 4: Add triethylamine (μL, 0.1 mmol) 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), and then add the product obtained in Step 3 (40.3 mg, 0.1 mmol). Stir the reaction mixture overnight at room temperature to obtain the ionizable lipid VC-LC-1194, and check the completion by TLC chromatography. Purify the obtained product by column chromatography using a gradient of dichloromethane / dichloromethane:methanol:ammonium hydroxide (80:20:1) (100 / 0 to 0 / 100). Combine the fractions containing the product, evaporate under reduced pressure to obtain a light yellow oil (yield: 67%).

[0426]

[0427] VC-LC-1194 was characterized by HPLC-ELSD-MS and the analytes were successfully detected using an evaporative light scattering detector and a simple quadrupole mass detector in the positive mode with a spectrophotometer Waters Acquity QDa.

[0428] Measured retention time: 13.28 minutes, MS(ES): experimental value m / z [M+H]+ 846.87 {theoretical value m / z [M+H]+ 846.64}.

[0429] HPLC-ELSD-MS analysis method

[0430] (HPLC was Water Alliance ELSD 2424, column was X bridge BEH C8, 4.6 mm, 50 mm, 2.5 μm, gradient was 5% to 95% B (A: water containing 0.01% TFA; B: acetonitrile containing 0.01% TFA), for 20 min, rate = 1 mL / min). Injection volume was 2 μL and column temperature was 65 °C.

[0431] Gradient: (A: water containing 0.01% TFA; B: acetonitrile containing 0.01% TFA), 30 min, constant flow rate = 1 mL / min)

[0432] Example 6 - Preparation and Characterization of LNPs Containing the Compounds of the Present Disclosure

[0433] Encapsulation of RNA in LNPs

[0434] In some cases, mRNA encoding the luciferase sequence containing SEQ ID NO: 1 in the 5' to 3' direction was encapsulated into LNPs by microfluidics or manually.

[0435] The microfluidic mixer uses the same procedure as described in Hassett, K. J., et al., "Optimization of Lipid Nanoparticles for Intramuscular Administration of mRNA Vaccines", 2019, Mol. Ther. Nucleic Acid, vol. 15, pp. 1 - 11. Briefly, the purified mRNA was first diluted to a final concentration of 266 μg / ml in sodium citrate buffer at pH = 4. Additionally, a mixture of the ionizable lipid of the present invention: DSPC (Merk 850365P): cholesterol (Sigma C3045): DMG-PEG2000 (Cayman 33945-1) was dissolved in ethanol at corresponding molar percentages of 50:10:38.5:1.5, and where the lipid nitrogen-to-phosphorus ratio (N:P) was 5.5:1.

[0436] Alternatively, when obtaining the LNP by manual methods, such as by 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), 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 LNP was immediately diluted 1:1 with Tris buffer and dialyzed overnight against Tris buffer containing 15% sucrose. Then the resulting LNP solution was collected and the encapsulated mRNA was evaluated by Ribogreen (Invitrogen R11490).

[0437] Then the LNP solution was adjusted to a final concentration of mRNA of 100 μg / ml. The particle size distribution, polydispersity, and ζ potential were measured by dynamic light scattering (DLS) using a Malvern Zetasizer Advance Lab Blue Label. RNA encapsulation was performed according to the manufacturer's instructions by Evaluate Ribogreen.

[0438] Finally, pass the LNP solution through a 0.22 mm filter and store the LNP at -80 °C until needed.

[0439] Table 6 presents the results of several parameters of LNPs containing specific ionizable lipids shown in the following standard formulations:

[0440] Ionizable lipid: non-cationic lipid: sterol: PEG-modified lipid → 50:10:38.5:1.5.

[0441] Table 6

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449] As shown in Table 6, each tested compound has an acceptable particle size for the purpose sought. The encapsulation efficiency, measured as the percentage of RNA encapsulated in the LNP, ranges from 100% to approximately 15%. However, this only indicates the ease with which the components of the nanoparticles interact to form LNPs capable of encapsulating RNA. It does not indicate the intracellular transfection efficiency. The intracellular transfection efficiency of mRNA (encoding luciferase expression) is measured by luminescence given as the total flux (p·s -1 ). The total flux is the bioluminescence measurement in photons per second or the average radiant rate in each pixel integrated over the target area.

[0450] As an illustrative example, the LNP containing compound VC-LC-0550 showed an encapsulation efficiency of 26.96%, but the measured total flux was 3.4·10 7 p·s -1 . This means that approximately 27% of the RNA is encapsulated when the RNA contacts the components of the LNP. The observed total flux value indicates that the LNP does not degrade immediately in vivo after injection and that the transfection of the cells is indeed very successful, so that the inclusions encapsulated in the LNP effectively reach the cytosol after endosomal escape.

[0451] Example 7 - Protein Expression in Mouse Muscle of Selected Instances of LNPs Prepared with the Lipids of the Invention and Containing mRNA as the Active Ingredient

[0452] Administration of mRNA (LNP) to Mice

[0453] Female BALB / c mice (Charles River Laboratories) aged 8 - 10 weeks and weighing 18 - 23 g were acclimated to the new conditions for 3 - 7 days after arrival at the experimental facility. The housing conditions were a room temperature of 20 - 24 °C, a humidity of 50 - 70%, a light intensity of 60 lux, and a light - dark cycle of 12 hours.

[0454] For measurement of firefly luciferase activity in mice, LNPs generated as described above (ionizable lipid:helper lipid:sterol:PEG - modified lipid → 50:10:38.5:1.5) and containing 1 μg of the designated mRNA in a final volume of 30 μl were injected intramuscularly.

[0455] At 4 to 72 hours after RNA - LNP inoculation, the mice were anesthetized by inhalation of 4% isoflurane using a vaporizer. Maintenance of anesthesia was carried out at 1.5% isoflurane. Then, D - luciferin (Quimigen, 12507) was injected intraperitoneally at a dose of 150 mg / kg, usually a 15 mg / mL stock solution in ∼200 μL of PBS for a 20 g mouse. Ten minutes after luciferin inoculation, luciferase images were acquired using an IVIS LuminaXRMS imaging system according to the manufacturer's instructions.

[0456] The following Table 7 (A - C) shows selected instances of LNPs containing the ionizable lipid of formula (I), the ionizable lipid containing the same polar head (morpholino) and different combinations of R2 and R3, to demonstrate that the general structure of formula (I) produces the desired effects independent of the alkyl residues. Figure 1 Protein expression in mice of the compounds described in Table 7 (A - C) is described.

[0457]

[0458]

[0459]

[0460] Table 8 shows selected examples of LNPs containing ionizable lipids of formula (I), said ionizable lipids containing various R3 substituents (polar heads) and different combinations of R2 and R3, to demonstrate the general structure of formula (I), in particular that the compounds of formula (II) produce the desired effects independent of the substituents R1, R2 and the selected polar head (R3).

[0461] Table 8

[0462]

[0463]

[0464]

[0465]

[0466]

[0467]

[0468]

[0469]

[0470]

[0471]

[0472]

[0473]

[0474]

[0475] Tables 7A - C and Table 8 confirm that LNPs containing ionizable lipids of formula II effectively encapsulate polynucleotides and produce high levels of cell transfection in vivo after administration.

[0476] Selected examples of LNPs containing ionizable lipids of formula (III) or ionizable lipids of formula (IV) and encapsulating luciferase - encoding mRNA were also tested in vivo. The results are shown in Table 9.

[0477] Table 9

[0478]

[0479]

[0480]

[0481]

[0482] The results summarized in Table 9 confirm that LNPs containing ionizable lipids of Formula III and Formula IV effectively encapsulate polynucleotides and produce high levels of cellular transfection in vivo after administration.

[0483] Example 8 - Immunoassays in Mice

[0484] Administration of mRNA-LNP to mice in different formulations

[0485] Female BALB / c mice (Charles River Laboratories), 8 - 10 weeks old and weighing 18 - 23 g, were acclimated to the new conditions for 3 - 7 days after arrival at the experimental facility. The housing conditions were a room temperature of 20 - 24 °C, humidity of 50 - 70%, light intensity of 60 lux, and a 12 - hour light - dark cycle.

[0486] For serum IgG - RBD measurements in mice, a therapeutically suitable composition was intramuscularly injected, the composition containing an LNP produced 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. The formulation is outlined in Table 10. The mRNA containing the sequence encoding SEQ ID NO: 2 in the 5' to 3' direction was encapsulated as previously disclosed in the present disclosure.

[0487] Table 10

[0488]

[0489] Blood was collected (submandibular bleeding) on day 21, and the blood samples were centrifuged at 10,000 rpm for 10 minutes to assay for serum antibodies by ELISA. The results are summarized in Table 11 below.

[0490] Table 11

[0491]

[0492] In all cases, the analysis by ELISA measurement of serum antibodies gave positive results. Samples IM - 3, IM - 4, IM - 5, and IM - 6 formulated with the ionizable lipids of Formula (I), particularly VC - LC - 0474, VC - LC - 0729, VC - LC - 0743, and VC - LC - 0867, gave better results compared to Control 1 and Control 2 corresponding to commercial vaccine formulations.

[0493] In each case, the mice were independently administered a booster that was identical to the formulation administered previously, and a second analysis was performed to examine the efficiency of the booster. The booster was administered 21 days after the first dose.

[0494] On day 21, blood was drawn from the submandibular (facial) vein. Blood samples on day 42 were obtained from euthanized mice.

[0495] The mice were euthanized by trained personnel using appropriate techniques, equipment, and reagents. The method chosen was inhalation of carbon dioxide (CO2) in the chamber, and it complied with the European Union Directive 2010 / 63 / UE on the care and use of laboratory animals in research. The research protocol was approved by the Animal Research Ethics Committee of the Universidad de Zaragoza.

[0496] The results are shown in Table 12 below.

[0497] Table 12

[0498]

[0499] As outlined in the table, in all cases, the formulations containing the ionizable lipid of formula (I) as described in the present disclosure obtained significantly higher results in the serum IgG-RBD assay, while for S-specific IFNγ-producing cells, IM-1, IM-2, and IM-3 obtained results comparable to those of Control-1 and Control-2, and IM-4, IM-5, and IM-6 obtained very good results.

[0500] According to the World Health Organization (WHO), the description of SEQ ID NO: 2 (CAS number: 2417899-77-3, messenger RNA encoding the full-length SARS-CoV-2 spike glycoprotein) is as follows:

[0501] CAP / 5'-UTR / sig / S protein mutant / 3'-UTR / poly(A)

[0502] UTR = untranslated region; sig = extended signal sequence of the S glycoprotein; S protein mutant = S glycoprotein sequence containing the mutations K986P and V987P; poly(A) = polyadenylation signal tail.

[0503]

[0504] 5'-cap structure 1-methyl-3'-pseudouridylyl cap G 1 A 2 = m 7 G + m 3' -5'-ppp-5'-Am 2' -3'-p-

[0505] [m 7 = 7-CH3; m 3' = 3'-O-CH3; m 2' = 2'-O-CH3;

[0506] -ppp- = -PO2H-O-PO2H-O-PO2H)-; -p- = -PO2H-]

[0507] Characteristic table

[0508]

[0509]

[0510]

[0511]

[0512]

[0513] Example 9 - Comparative example

[0514]

[0515] The compound VC-LC-0588 depicted herein was synthesized by a thiolactone ring-opening reaction in a combinatorial library of 288 lipid compounds for gene delivery applications according to the same synthetic method previously reported by Molla et al. (Bioconjugate Chem. 2018, 29, 4, 992-999, referred to as T18U-PY12-A), to obtain a compound containing an unsaturated bond and a reducible disulfide bond. The study showed the results of testing the compound in vitro using HEK293T cells without adding co-lipids. The authors reported highly stable liposomes with low toxicity and a transfection efficiency of approximately 95%. However, the study did not report in vivo experiments in mammals. Therefore, in this article, in vivo studies were also conducted on VC-LC-0588 to compare the differences between compounds of general formula (I) containing thioether instead of disulfide groups and the effect of at least one ester group near the S atom in the thioether. As Figure 2 shown in B, after intramuscular injection of LNP encapsulating mRNA encoding luciferase and containing VC-LC-0588 as an ionizable lipid of the following formula, no transfection (luminescence) was observed in vivo:

[0516] Ionizable lipid: Co-lipid: Sterol: PEGylated lipid → 50:10:38.5:1.5.

[0517] The compounds depicted in Table 13 were also synthesized, characterized, and tested in vivo. These compounds are partially similar to the dimers of formula (I), but a disulfide moiety is introduced, thus showing the general formula depicted herein:

[0518]

[0519] wherein A, B, n, R1, and R3 are the same as those defined in general formula (I).

[0520] Table 13

[0521]

[0522] As Figure 2 shown in A, after intramuscular injection of an LNP containing VC-LC-0236 of the following formula as an ionizable lipid and encapsulating mRNA encoding luciferase, no transfection (luminescence) was observed in vivo: ionizable lipid: helper lipid: sterol: polyethylene glycolylated lipid → 50:10:38.5:1.5.

[0523] Example 10 - Optimization of Formulations

[0524] Compound VC-LC-0431 was selected to test different composition ratios.

[0525] As previously shown in Tables 7A-C and 8 (Example 7), for the ionizable lipids of formula (I) of the present invention, in vivo tests were performed on a standard formulation having the following composition ratio: ionizable lipid: helper lipid: sterol: polyethylene glycolylated lipid → 50:10:38.5:1.5.

[0526] wherein, as described above, the ionizable lipid is VC-LC-0431, the helper lipid is DOPE, the sterol is cholesterol, and the polyethylene glycolylated lipid is DMG-PEG2000.

[0527] The following Table 14 shows the results obtained for the said standard composition:

[0528] Table 14

[0529]

[0530] Optimization of LNP Formulations

[0531] Different ratios of the components included in the mixture forming the LNP were tested to obtain an optimized formulation, thereby improving the in vivo transfection rate.

[0532] The test compositions depicted in Table 15 were intraperitoneally injected into mice, and significant improvements were observed for some formulations.

[0533] Table 15

[0534]

[0535] The results depicted in Table 15 confirm that by changing the composition ratio of the components in the LNP formulation of the ionizable lipid of formula (I) of the present invention, the in vivo transfection rate can be increased. When compared with the previous LNP with a standard composition of ionizable lipid:DOPE:cholesterol:DMG-PEG2000 of 50:10:38.5:1.5, the LNP of Compositions 3-8 achieved a higher transfection rate in vivo. Figure 3 In vivo transfection in mice after administration of the 8 compositions defined in Table 15 is depicted.

[0536] Example 11. Optimized Formulations with High Cholesterol Percentage

[0537] Using two different ionizable lipids included in the mixture for forming LNP, optimized compositions with a high cholesterol percentage and without co-lipids (especially phospholipids) were tested. The in vivo transfection rate was tested using these formulations.

[0538] The test compositions outlined in Table 16 below were intraperitoneally injected into mice, and significantly high transfection efficiency (measured by total flux) was observed.

[0539] Table 16

[0540]

[0541] Example 12 - Stability of LNP over Time

[0542] Liquid sample stored at T = 4 °C

[0543] Two LNPs were tested to determine their stability. The LNPs contained: an ionizable lipid (formula I) in a ratio of 50:10:38.5:1.5: non-cationic lipid:cholesterol:PEG-modified lipid, and an mRNA containing a sequence encoding luciferase of SEQ ID NO.1 in the 5' to 3' direction. Encapsulation was carried out as previously described in the present disclosure.

[0544] Formulation A containing VC-LC-0163 as the ionizable lipid was administered to mice, and in the first experiment, fresh formulation was used, and the transfection efficiency was measured using the formulation that had been stored at T = 4°C for 83 days.

[0545] Formulation B containing VC-LC-0263 as the ionizable lipid was administered to mice, and in the first experiment, fresh formulation was used, and the transfection efficiency was measured using the formulation that had been stored at T = 4°C for 61 days. The results are summarized in Table 17 below:

[0546] Table 17

[0547]

[0548] The results obtained confirm that after long-term storage under normal refrigeration conditions at 4 °C, the LNPs containing the ionizable lipids of formula (I) of the present disclosure are still capable of generating in vivo transfection.

[0549] Effect of storage conditions on in vivo immune studies. Comparative example: Fresh solution stored at -80 °C compared to lyophilized preparations stored at 4 °C and 25 °C

[0550] Two different mRNA-encapsulating LNPs were tested to determine whether storage conditions might have an impact on in vivo immunity. Thus, a commercial LNP (LNPA) was compared with another LNP (LNG B) formulated with the ionizable lipid of formula (I) described in the present disclosure (specifically, the ionizable lipid VC-LC-0729, 2-hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate). After storage of LNPA and LNP B in solution at T = -80 °C, or after lyophilization at a temperature of 4 °C in one case and 25 °C in another case, they were tested in vivo (in mice). Both LNPA and LNPB were prepared by standard microfluidic mixing as disclosed previously in Hassett, K.J. et al. which has been defined in the present disclosure. The lyophilization of the LNPs can be carried out by a person skilled in the art by standard methods known in the art. For example, a helpful 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).

[0551] For serum IgG-RBD measurements in mice, a therapeutically suitable composition containing LNPA or B produced as described above (ionizable lipid: helper lipid: sterol: conjugated or PEGylated lipid) and containing 1 μg of the designated mRNA in a final volume of 30 μl was intramuscularly injected. At 21 days after administration, blood was collected by submandibular bleeding, and the samples were centrifuged at 10,000 rpm for t = 10 min to determine serum antibodies by ELISA, and the results are shown in Table 18 below.

[0552] The compositions of LNPA and LNP B are described herein:

[0553] LNP A contains the mRNA sequence SEQ ID NO:: ALC-0315:DSPC:cholesterol:ALC-0159-46.3:9.4:42.7:1.6 (% mol) encapsulated in a lipid formulation as described below.

[0554] ALC-0315 and ALC-0159 are commercially available (ALC-0315 CAS number: 2036272-55-4, ALC-0159 CAS number: 1849616-42-7).

[0555] LNP B contains the mRNA sequence SEQ ID NO: 3: VC-LC-0729:DOPE:cholesterol:DMG-PEG2000-40.7:34.9:23.3:1.2 (% mol) encapsulated in a lipid formulation as described below.

[0556] Encapsulation of the mRNA comprising the sequence of SEQ ID NO: 2 or SEQ ID NO: 3 in the 5' to 3' direction is carried out as previously described in this disclosure.

[0557] SEQ ID NO.: 3

[0558]

[0559]

[0560]

[0561] Ψ = 1-methyl-3'-pseudouridinyl

[0562] Table 18

[0563]

[0564] When stored in solution at -80 °C, LNPA performs better than LNP B at T = 0 days. However, for T = 90 days, the results obtained are comparable.

[0565] Notably, for the lyophilized LNPs stored for a 90-day period at 4 °C or 25 °C, the IgGRBD measurements of LNPB are much better than those of LNPA lyophilized and stored under the same conditions.

[0566] Effect of storage conditions and re-suspended lyophilized samples over time on in vivo immunological studies

[0567] The LNP was lyophilized according to standard protocols known in the art as described above.

[0568] After lyophilization, the sample was immediately resuspended in the same buffer, namely Tris 20 mM + 15% sucrose with 5% trehalose added, and immediately stored in a refrigerator at T = 4 °C.

[0569] Immunological studies were performed 24 h and 96 h after sample resuspension. For serum IgG-RBD measurements in mice, a therapeutically suitable composition of LNP (A and B) containing 1 μg of the designated mRNA in a final volume of 30 μl was intramuscularly injected into the mice. Blood was collected by submandibular bleeding on day 21, and the samples were centrifuged at 10,000 rpm for t = 10 min to determine serum antibodies by ELISA. The results are summarized in Table 19 below.

[0570] Table 19

[0571]

[0572] After 24 hours or 96 hours of resuspension, lyophilized LNP B showed better immunological results than commercial LNP A; although in both cases the values obtained after 96 hours were higher (in fact, this is the result of the variability between subjects), when compared in parallel, it is clear that after 24 hours of resuspension, the values obtained for LNP B were significantly higher than those for LNP A (LNP B was 54% higher), and the same was observed for the results obtained after 96 hours of resuspension (LNP B was 38% higher).

[0573] Example 13 - Effect of Different Encapsulation Methods on Transfection Efficiency

[0574] For some 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, VC-LC-0881, the effect of the synthesis method used to obtain the LNP on transfection efficiency was evaluated.

[0575] Administration of mRNA(LNP) to mice, two different therapeutic doses.

[0576] Female BALB / c mice (Charles River Laboratories) at 8 - 10 weeks of age and weighing 18 - 23 g were acclimated to the new conditions for 3 - 7 days after arrival at the experimental facility. The housing conditions were a room temperature of 20 - 24 °C, a humidity of 50 - 70%, a light intensity of 60 lux, and a light - dark cycle of 12 hours.

[0577] For the measurement of firefly luciferase activity in mice, LNPs generated as described above (ionizable lipid:helper lipid:sterol:PEG - modified lipid → 50:10:38.5:1.5) and containing 1 μg of the designated mRNA in a final volume of 30 μl or 0.5 μg of the designated mRNA in a final volume of 30 μl were injected intramuscularly.

[0578] At 4 to 72 hours after RNA - LNP inoculation, the mice were anesthetized by inhalation of 4% isoflurane using a vaporizer. Maintenance of anesthesia was carried out at 1.5% isoflurane. Then, D - luciferin (Quimigen, 12507) was injected intraperitoneally at a dose of 150 mg / kg, which is typically a 15 mg / mL stock solution in ~200 μL of 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.

[0579] In the primary screening, the expression of luciferase mRNA encapsulated in LNPs formulated with different ionizable lipids having a standard ratio of the following molar ratios (ionizable lipid / helper lipid / sterol / PEG - lipid): 50 / 10 / 38.5 / 1.5 was evaluated in vivo. The LNPs were manually synthesized in 96 - well plates.

[0580] In the secondary screening, the expression of luciferase mRNA encapsulated in LNPs formulated with different ionizable lipids having a standard ratio of the following molar ratios (ionizable lipid / helper lipid / sterol / PEG - lipid): 50 / 10 / 38.5 / 1.5 was evaluated. The LNPs were manually synthesized again in 96 - well plates to confirm the consistency of the results and were also synthesized by microfluidics in an Ignite device (Precision Nanosystems); two repeated measurements were performed on the LNPs obtained by microfluidics, and the results are shown in Table 20 below.

[0581] Table 20

[0582]

[0583]

[0584]

[0585]

[0586] The results obtained in the primary screening showed that the in vivo transfection efficiency results measured by total flux were very good. For example, for LNPs containing VC-LC-0729 as the ionizable lipid (1 μg mRNA in a total volume of 30 μL), it was as high as 3.61E+08.

[0587] Overall, the secondary screening improved the transfection efficiency observed in mice. For example, the observed values were as high as 1.37E+09 for VC-LC-0729, or as high as 1.34E+09 for VC-LC-10743 (1 μg mRNA in a total volume of 30 μL).

[0588] Example 14. Effect of in vivo injection dose on transfection efficiency

[0589] 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 compared to 0.5 μg mRNA in a total volume of 30 μL) were tested in the primary and secondary screenings in 96-well plates.

[0590] As shown in Table 21 below, for the results obtained in the primary screening at full dose and half dose, VC-LC-0866 and VC-LC-0868 were comparable, VC-LC-0867 was slightly lower, and improvements were observed for VC-LC-0872 and VC-LC-0881.

[0591] Table 21

[0592]

[0593]

[0594] The results shown in Table 21 indicate that even when half the dose was injected in mice, the observed transfection efficiency (measured by total flux) was very high, ranging from 1.06E+08 to 2.02E+08. Therefore, the ionizable lipids of the present invention included in LNPs containing therapeutic agents such as mRNA can successfully achieve high transfection rates even at lower doses.

[0595] The present invention includes the following clauses:

[0596] The first group of clauses:

[0597] 1. An ionizable lipid of formula (II):

[0598]

[0599] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0600] A is

[0601] B is

[0602] D is

[0603] n, m and p are independently 0, 1, 2, 3, 4, 5 or 6;

[0604] q is selected from 1, 2 and 3;

[0605] t is selected from 0, 1 and 2;

[0606] and wherein q + t = 1, 2 or 3, provided that when q + t = 1, then when R1 is a straight-chain alkyl substituent, R1 contains at least 7 carbon atoms;

[0607] R1 and R2 are independently straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, wherein R1 and R2 are optionally substituted with one or more substituents selected from -OH, -COOR4 and -C(=O)S-R4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl,

[0608] and wherein

[0609] R3 is a heterocycle containing at least one N atom, or R3 is:

[0610]

[0611] wherein Ra and Rb are independently straight-chain or branched C1-C6 alkyl optionally substituted with a hydroxyl group.

[0612] 2. The ionizable lipid according to clause 1, 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.

[0613] 3. The ionizable lipid according to clause 1 or 2, wherein

[0614] q is selected from 1 and 2, t is selected from 0 and 1, and wherein q + t = 2;

[0615] or

[0616] wherein q is selected from 1 and 2, t is selected from 0 and 1, q + t = 2; and R3 is a 5-membered or 6-membered ring containing one N atom and an optional second heteroatom preferably selected from N and O;

[0617] or

[0618] where q is selected from 1 and 2, t is selected from 0 and 1, and q + t = 2; and R3 is selected from the following structures:

[0619]

[0620] 4. The ionizable lipid according to any one 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.

[0621] 5. The compound according to clause 4, wherein n is 2 or 3, m = t = 0, p = q = 2,

[0622]

[0623] wherein

[0624] R1 is a C10-C20 branched alkyl;

[0625] R2 is a C2-C30 straight or branched alkyl or a C6-C24 straight alkenyl or alkynyl; and

[0626] R3 is:

[0627]

[0628] where Ra = Rb = C1-C4 alkyl;

[0629] or wherein n = 2, R2 is a C2-C30 straight or branched alkyl, and wherein R3 is:

[0630] where Ra = Rb = C1-C4 alcohol-terminated alkyl;

[0631] or

[0632] wherein n is 2 or 3, m = t = 0, p = q = 2,

[0633] A is

[0634] where R1 is a C10-C20 branched alkyl, R2 is a C3-C30 straight or branched alkyl, and wherein R3 is an imidazole ring;

[0635] or

[0636] wherein n = 3, m = t = 0, p = q = 2,

[0637] A is

[0638] wherein R1 is a C10-C20 branched alkyl group, R2 is a C4-C20 straight-chain or branched alkyl group, and wherein R3 is a pyrrolidine ring;

[0639] or

[0640] wherein n = 2, m = t = 0, p = q = 2,

[0641] A is

[0642] wherein R1 is a C10-C20 straight-chain or branched alkyl group, R2 is a C4-C20 straight-chain or branched alkyl group, and wherein R3 is N-4-methylpiperazine;

[0643] or

[0644] wherein n is selected from 2 or 3,

[0645] m = t = 0 and p = q = 2,

[0646]

[0647] R1 is a C10-C22 straight-chain or branched alkyl, alkenyl or alkynyl group; R2 is a C3-C30 straight-chain or branched alkyl group or a C3-C30 straight-chain alkenyl or alkynyl group; and wherein R3 is a morpholine ring;

[0648] or

[0649] wherein n is selected from 2 or 3, m = 0, p = 2,

[0650] q = t = 1,

[0651] A is

[0652] B = D, R1 is a C10-C22 straight-chain branched alkyl group; R2 is a C3-C30 straight-chain or branched alkyl group or a C3-C30 straight-chain alkenyl or alkynyl group, and R3 is a morpholine ring;

[0653] or

[0654] wherein n = q = p = 2; t = m = 0;

[0655] A is

[0656] B is

[0657] D is

[0658] R1 is a C10-C22 branched alkyl group;

[0659] R2 is a C8-C20 linear alkenyl or alkynyl group; or a C3-C28 linear alkyl group optionally substituted with one or more substituents selected from OH, -COOR4, and -COSR4, where R4 is a C1-C6 alkyl group, a C2-C6 alkenyl group, or a C2-C6 alkynyl group; or a C12-C25 branched alkyl group;

[0660] and R3 is -N(CH3)2 or

[0661] 6. The ionizable lipid according to Clause 1, which is a compound selected from the following: 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, as defined in the specification.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-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, 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, or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them.,

[0662] 7. A lipid nanoparticle comprising an ionizable lipid according to any one of clauses 1 to 6.

[0663] 8. The lipid nanoparticle according to clause 7, further comprising:

[0664] i) a non-cationic lipid;

[0665] Optionally, the non-cationic lipid is selected from: 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-docosanoyl-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-cholesteryl succinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16Lyso 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 16.0PE), 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;

[0666] Optionally, the non-cationic lipid is DOPE; and / or the non-cationic lipid is DSPC;

[0667] ii) sterol;

[0668] Optionally, the sterol is selected from cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof;

[0669] Further optionally, the sterol is cholesterol; and

[0670] iii) PEG-modified lipid;

[0671] Optionally, the PEG-modified lipid is selected from PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified phosphatidylcholine, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

[0672] 9. The lipid nanoparticle according to any one of clauses 7 to 8, further comprising a non-cationic lipid, a sterol, and a PEG-modified lipid, wherein the amount of the ionizable lipid is 25 to 60 mol%, the amount of the PEG-modified lipid is 0.1 to 10 mol%; the amount of the non-cationic lipid is 10 to 45 mol%; and the amount of the sterol is 10 to 40 mol%.

[0673] 10. The lipid nanoparticle according to any one of clauses 7 to 9, further comprising a pharmaceutically active agent.

[0674] 11. The lipid nanoparticle according to clause 10, wherein the pharmaceutically active agent is selected from polynucleotides, DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide, and expression vectors containing DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide.

[0675] 12. The lipid nanoparticle according to clause 11, wherein the polynucleotide is natural or artificial deoxyribonucleic acid (DNA) or natural or artificial ribonucleic acid (RNA);

[0676] Further optionally, the polynucleotide comprises at least one chemical modification 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-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine (also known as m5U), 5-methoxyuridine, 2'-O-methyluridine, and combinations thereof. In particular, the chemical modification is N1-methylpseudouridine, 5-methoxyuridine, or a combination thereof; in particular, the chemical modification is N1-methylpseudouridine;

[0677] Further optionally, the polynucleotide is ribonucleic acid (RNA);

[0678] Further optionally, wherein the RNA is selected from 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), small hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof; in particular, the RNA is mRNA.

[0679] 13. A pharmaceutical composition comprising the lipid nanoparticles according to any one of clauses 10 to 12 and a pharmaceutically acceptable excipient or carrier.

[0680] 14. The lipid nanoparticles according to any one of clauses 10 to 12 or the pharmaceutical composition according to clause 13, for use in a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition;

[0681] Optionally, wherein the disease or disorder is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular and renal vascular diseases, and metabolic diseases;

[0682] Further optionally, wherein the subject is a human;

[0683] Or for use in a method of inducing an immune response in a subject, a method of therapeutic immunization for a subject, for use as a vaccine, or for gene therapy;

[0684] Or for preventing or treating COVID-19.

[0685] 15. Use of the lipid nanoparticles according to any one of clauses 7 - 9 as an encapsulant.

[0686] Second set of clauses:

[0687] 1. An ionizable lipid of formula (II):

[0688]

[0689] Or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0690] A is

[0691] B is

[0692] D is

[0693] n, m, and p are independently 0, 1, 2, 3, 4, 5, or 6;

[0694] q is selected from 1, 2, and 3;

[0695] t is selected from 0, 1, and 2;

[0696] and wherein q + t = 1, 2, or 3, provided that when q + t = 1, then when R1 is a straight-chain alkyl substituent, R1 contains at least 7 carbon atoms;

[0697] R1 and R2 are independently straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, wherein R1 and R2 are optionally substituted by one or more substituents selected from -OH, -COOR4, and -C(=O)S-R4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl,

[0698] and wherein

[0699] R3 is a heterocycle containing at least one N atom, or R3 is:

[0700]

[0701] wherein Ra and Rb are independently straight-chain or branched C1-C6 alkyl optionally substituted by hydroxy.

[0702] 2. The ionizable lipid according to clause 1, 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.

[0703] 3. The ionizable lipid according to clause 1 or 2, wherein

[0704] q is selected from 1 and 2, t is selected from 0 and 1, and wherein q + t = 2;

[0705] or

[0706] wherein q is selected from 1 and 2, t is selected from 0 and 1, q + t = 2; and R3 is a 5-membered or 6-membered ring containing one N atom and an optional second heteroatom preferably selected from N and O;

[0707] or

[0708] wherein q is selected from 1 and 2, t is selected from 0 and 1, q + t = 2; and R3 is selected from the following structures:

[0709]

[0710] 4. An ionizable lipid according to any one 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.

[0711] 5. The compound according to clause 4, wherein n is 2 or 3, m = t = 0, p = q = 2,

[0712]

[0713] wherein

[0714] R1 is a C10-C20 branched alkyl;

[0715] R2 is a C2-C30 straight or branched alkyl or a C6-C24 straight alkenyl or alkynyl; and

[0716] R3 is:

[0717] where Ra = Rb = C1-C4 alkyl;

[0718] or wherein n = 2, R2 is a C2-C30 straight or branched alkyl, and wherein R3 is:

[0719] where Ra = Rb = C1-C4 alcohol-terminated alkyl;

[0720] or

[0721] wherein n is 2 or 3, m = t = 0, p = q = 2,

[0722] A is

[0723] where R1 is a C10-C20 branched alkyl, R2 is a C3-C30 straight or branched alkyl, and wherein R3 is an imidazole ring;

[0724] or

[0725] wherein n = 3, m = t = 0, p = q = 2,

[0726] A is

[0727] where R1 is a C10-C20 branched alkyl, R2 is a C4-C20 straight or branched alkyl, and wherein R3 is a pyrrolidine ring;

[0728] or

[0729] wherein n = 2, m = t = 0, p = q = 2,

[0730] A is

[0731] wherein R1 is a C10-C20 straight-chain or branched alkyl group, R2 is a C4-C20 straight-chain or branched alkyl group, and wherein R3 is N-4-methylpiperazine;

[0732] or

[0733] wherein n is selected from 2 or 3,

[0734] m = t = 0, and p = q = 2,

[0735]

[0736] R1 is a C10-C22 straight-chain, branched alkyl, alkenyl or alkynyl group; R2 is a C3-C30 straight-chain or branched alkyl group or a C3-C30 straight-chain alkenyl or alkynyl group; and wherein R3 is a morpholine ring;

[0737] or

[0738] wherein n is selected from 2 or 3, m = 0, p = 2,

[0739] q = t = 1,

[0740] A is

[0741] B = D, R1 is a C10-C22 straight-chain branched alkyl group; R2 is a C3-C30 straight-chain or branched alkyl group or a C3-C30 straight-chain alkenyl or alkynyl group, and R3 is a morpholine ring;

[0742] or

[0743] wherein n = q = p = 2; t = m = 0;

[0744] A is

[0745] B is

[0746] D is

[0747] R1 is a C10-C22 branched alkyl group;

[0748] R2 is a C8-C20 straight-chain alkenyl or alkynyl group; or a C3-C28 straight-chain alkyl group optionally substituted with one or more substituents selected from OH, -COOR4 and -COSR4, wherein R4 is a C1-C6 alkyl group, a C2-C6 alkenyl group or a C2-C6 alkynyl group; or a C12-C25 branched alkyl group;

[0749] and R3 is -N(CH3)2 or

[0750] 6. The ionizable lipid according to Clause 1, which is a compound selected from the following: 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, as defined in the specification.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-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, 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, or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them.,

[0751] 7. A lipid nanoparticle comprising an ionizable lipid according to any one of clauses 1 to 6.

[0752] 8. The lipid nanoparticle according to clause 7, further comprising:

[0753] i) a non-cationic lipid;

[0754] Optionally, the non-cationic lipid is selected from: 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-docosanoyl-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-cholesteryl succinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16Lyso 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 16.0PE), 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;

[0755] Optionally, the non-cationic lipid is DOPE; and / or the non-cationic lipid is DSPC;

[0756] ii) sterols;

[0757] Optionally, the sterol is selected from cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof;

[0758] Further optionally, the sterol is cholesterol; and

[0759] iii) PEG-modified lipids;

[0760] Optionally, the PEG-modified lipid is selected from PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified phosphatidylcholine, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

[0761] 9. The lipid nanoparticle according to any one of clauses 7 to 8, further comprising a non-cationic lipid, a sterol, and a PEG-modified lipid, wherein the amount of the ionizable lipid is 25 to 60 mol%, the amount of the PEG-modified lipid is 0.1 to 10 mol%; the amount of the non-cationic lipid is 10 to 45 mol%; and the amount of the sterol is 10 to 40 mol%.

[0762] 10. The lipid nanoparticle according to any one of clauses 7 to 9, further comprising a pharmaceutically active agent.

[0763] 11. The lipid nanoparticle according to clause 10, wherein the pharmaceutically active agent is selected from polynucleotides, DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide, and expression vectors containing DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide.

[0764] 12. The lipid nanoparticle according to clause 11, wherein the polynucleotide is natural or artificial deoxyribonucleic acid (DNA) or natural or artificial ribonucleic acid (RNA);

[0765] Further optionally, the polynucleotide comprises at least one chemical modification 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-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine (also known as m5U), 5-methoxyuridine, 2'-O-methyluridine, and combinations thereof. In particular, the chemical modification is N1-methylpseudouridine, 5-methoxyuridine, or a combination thereof; in particular, the chemical modification is N1-methylpseudouridine;

[0766] Further optionally, the polynucleotide is ribonucleic acid (RNA);

[0767] Further optionally, wherein the RNA is selected from 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), small hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof; in particular, the RNA is mRNA.

[0768] 13. A pharmaceutical composition comprising the lipid nanoparticle according to any one of clauses 10 to 12 and a pharmaceutically acceptable excipient or carrier.

[0769] 14. The lipid nanoparticle according to any one of clauses 10 to 12 or the pharmaceutical composition according to clause 13, for use in a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition;

[0770] Optionally, wherein the disease or disorder is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular and renal vascular diseases, and metabolic diseases;

[0771] Further optionally, wherein the subject is a human;

[0772] Or for use in a method of inducing an immune response in a subject, a method of therapeutic immunization for a subject, for use as a vaccine, or for gene therapy;

[0773] Or for preventing or treating COVID-19.

[0774] 15. Use of the lipid nanoparticle according to any one of clauses 7 - 9 as an encapsulant.

[0775] Third set of clauses:

[0776] 1. An ionizable lipid of formula (I):

[0777]

[0778] Or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0779] A is selected from -NR'-C(O)-, -C(O)-NR'-, -C(O)-O-, and -O-C(O)-, wherein R' is selected from H, methyl, and ethyl;

[0780] B is selected from -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -O-C(O)-;

[0781] n, m and p are independently selected from 0, 1, 2, 3, 4, 5 and 6;

[0782] q is selected from 1, 2 and 3;

[0783] t is selected from 0, 1 and 2;

[0784] and wherein q + t = 1, 2 or 3;

[0785] R3 is a heterocycle containing at least one N atom, or R3 is:

[0786]

[0787] wherein Ra and Rb are independently straight-chain or branched C1-C6 alkyl optionally substituted by hydroxy;

[0788] X is selected from -D-R2,

[0789] wherein j, j’ and j” are independently selected from 0, 1 and 2,

[0790] Z is selected from -R1 and

[0791] wherein f and f’ are independently selected from 0, 1, 2, 3, 4, 5 and 6,

[0792] wherein g and g’ are independently selected from 1, 2, 3, 4, 5 and 6,

[0793] wherein D is independently selected from -C(O)-O- or -O-C(O)-,

[0794] and wherein R1, R’1, R2, R’2 and R”2 are independently 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 by one or more substituents selected from -OH, -COOR4 and -C(=O)SR4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl.

[0795] 2. The ionizable lipid according to clause 1, wherein when q + t = 1, X = -D-R2, D = -O-C(O)-, A = -NH-C(O)-, B = -NH-C(O)-, Z = R1 and R1 is straight-chain or branched alkyl, the moiety -(CH2) m -R1 contains at least 7 carbon atoms.

[0796] 3. The ionizable lipid according to clause 1, wherein when n = t = m = 0, q = p = 1,

[0797] X is

[0798] wherein one of j and j' is 0 and the other is 1,

[0799] when D is -O-C(O)-, A = -C(O)-NH-, B = -NH-C(O)-, Z is R1 and R1 is a C1-C18 alkyl group,

[0800] R2 and R'2 are not C7-C18 alkyl groups.

[0801] 4. The ionizable lipid according to clause 1, which is a compound of formula (II):

[0802]

[0803] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A, B, D, n, m, p, q, t, q + t, R1 and R2 are as defined in clause 1,

[0804] or

[0805] wherein when q + t = 1, X = -D-R2, D = -O-C(O)-, A = -NH-C(O)-, B = -NH-C(O)-, Z = R1 and R1 is a straight-chain or branched alkyl group, the moiety -(CH2) m -R1 contains at least 7 carbon atoms.

[0806] 5. The ionizable lipid according to clause 4, wherein A is -NH-C(O)- or -C(O)-NH-.

[0807] 6. The ionizable lipid according to clause 4 or 5, wherein n is selected from 0, 1, 2 and 3; p is selected from 0, 1, 2 and 3; m is selected from 0, 1, 2 and 3.

[0808] 7. The ionizable lipid according to clauses 4 to 6, wherein

[0809] q is selected from 1 and 2, t is selected from 0 and 1, and wherein q + t = 2.

[0810] 8. The ionizable lipid according to any one of clauses 1 to 7, wherein R3 is a 5-membered or 6-membered ring containing one N atom and optionally a second heteroatom preferably selected from N and O.

[0811] 9. The ionizable lipid according to any one of clauses 1 to 8, wherein R3 is selected from the following structures:

[0812]

[0813] 10. An ionizable lipid according to any one of the preceding clauses 4-9, 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.

[0814] 11. The compound according to clause 10, wherein n is 2 or 3, m = t = 0, p = q = 2,

[0815]

[0816] wherein

[0817] R1 is a C10-C20 branched alkyl;

[0818] R2 is a C2-C30 straight-chain or branched alkyl or a C6-C24 straight-chain alkenyl or alkynyl; and

[0819] R3 is

[0820] where Ra = Rb = C1-C4 alkyl.

[0821] 12. The ionizable lipid according to clause 10, wherein n = 2, R2 is a C2-C30 straight-chain or branched alkyl, and wherein R3 is:

[0822]

[0823] where Ra = Rb = C1-C4 alcohol-terminated alkyl.

[0824] 13. The compound according to clause 10, wherein n is 2 or 3, m = t = 0, p = q = 2,

[0825] A is

[0826] where R1 is a C10-C20 branched alkyl, R2 is a C3-C30 straight-chain or branched alkyl, and wherein R3 is an imidazole ring.

[0827] 14. The ionizable lipid according to clause 10, wherein n = 3, m = t = 0, p = q = 2,

[0828] A is

[0829] where R1 is a C10-C20 branched alkyl, R2 is a C4-C20 straight-chain or branched alkyl, and wherein R3 is a pyrrolidine ring.

[0830] 15. The compound according to clause 10, wherein n = 2, m = t = 0, p = q = 2,

[0831] A is

[0832] wherein R1 is a C10-C20 straight-chain or branched alkyl group, R2 is a C4-C20 straight-chain or branched alkyl group, and wherein R3 is N-4-methylpiperazine.

[0833] 16. The ionizable lipid according to clause 10, wherein n is selected from 2 or 3,

[0834] m = t = 0, and p = q = 2,

[0835]

[0836] R1 is a C10-C22 straight-chain, branched alkyl, alkenyl or alkynyl group; R2 is a C3-C30 straight-chain or branched alkyl group or a C3-C30 straight-chain alkenyl or alkynyl group; and wherein R3 is a morpholine ring.

[0837] 17. The ionizable lipid according to clause 10, wherein n is selected from 2 or 3, m = 0, p = 2,

[0838] q = t = 1,

[0839] A is

[0840] B = D, R1 is a C10-C22 straight-chain branched alkyl group; R2 is a C3-C30 straight-chain or branched alkyl group or a C3-C30 straight-chain alkenyl or alkynyl group, and R3 is a morpholine ring.

[0841] 18. The ionizable lipid according to clause 10, wherein n = q = p = 2; t = m = 0;

[0842] A is

[0843] B is

[0844] D is

[0845] R1 is a C10-C22 branched alkyl group;

[0846] R2 is a C8-C20 straight-chain alkenyl or alkynyl group; or optionally a C3-C28 straight-chain alkyl group substituted with one or more substituents selected from OH, -COOR4 and -COSR4, wherein R4 is a C1-C6 alkyl group, a C2-C6 alkenyl group or a C2-C6 alkynyl group; or a C12-C25 branched alkyl group;

[0847] and R3 is -N(CH3)2 or

[0848] 19. The ionizable lipid according to clause 1, which is a compound of formula (III):

[0849]

[0850] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0851] A is -NH-C(O)- or -C(O)-NH-,

[0852] R1, R2 and R'2 are independently selected from straight-chain or branched C1-C30 alkyl, C2-C30 alkenyl, C2-C30 alkynyl, wherein each of R1, R2, R'2 is optionally substituted with one or more substituents selected from -OH, -COO(R4) and -COSR4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl; and

[0853] wherein B, D, n, m, p, q, t, q+t, j, j' and R3 are as defined in clause 1,

[0854] or

[0855] wherein when n = t = m = 0, q = p = 1, A = B = -NH-C(O)-,

[0856] wherein one of j and j' is 0 and the other is 1, D = -O-C(O)-, and when R1 is selected from C1-C18 alkyl, R2 and R'2 are not C7-C18 alkyl.

[0857] 20. The ionizable lipid according to clause 1, wherein the compound is a compound of formula (IV):

[0858]

[0859] or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein

[0860] A is -NH-C(O)- or -C(O)-NH-,

[0861] wherein B, D, n, m, p, q, t, q+t, f, f’, g, g’ and R3 are as defined in clause 1.

[0862] 21. The ionizable lipid according to Clause 1, which is a compound selected from the following group: 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, described herein.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-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, 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-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-1096VC-LC-1097, 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 a stereoisomer of any of them.

[0863] 22. A lipid nanoparticle comprising an ionizable lipid according to any one of clauses 1 to 21.

[0864] 23. The lipid nanoparticle according to clause 22, further comprising a non-cationic lipid;

[0865] Optionally, wherein the non-cationic lipid is selected from: 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-docosanoyl-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-cholesteryl succinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16Lyso 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 16.0PE), 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;

[0866] Optionally, wherein the non-cationic lipid is DOPE; and / or wherein the non-cationic lipid is DSPC.

[0867] 24. The lipid nanoparticle according to clause 22 or 23, further comprising a sterol or a sterol precursor;

[0868] Optionally, wherein the sterol is selected from cholesterol, coprostanol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof;

[0869] Further optionally, wherein the sterol is cholesterol.

[0870] 25. The lipid nanoparticle according to any one of clauses 22 to 23, further comprising a PEGylated lipid or a conjugated lipid;

[0871] Optionally, wherein the PEGylated lipid is selected from PEGylated phosphatidylethanolamine, PEGylated phosphatidic acid, PEGylated phosphatidylcholine, PEGylated ceramide, PEGylated dialkylamine, PEGylated diacylglycerol, PEGylated dialkylglycerol, and mixtures thereof.

[0872] 26. The lipid nanoparticle according to any one of clauses 22 to 24, comprising a non-cationic lipid, a sterol, and a PEGylated lipid, wherein the amount of the ionizable lipid is 25 to 60 mol%, the amount of the PEGylated lipid is 0.1 to 10 mol%; the amount of the non-cationic lipid is 10 to 45 mol%; and the amount of the sterol is 10 to 40 mol%;

[0873] Or

[0874] wherein the amount of the ionizable lipid is 25 to 64 mol%, the amount of the PEGylated lipid is 0.1 to 1.5 mol%; and the amount of the sterol is 35 to 74 mol%.

[0875] 27. The lipid nanoparticle according to any one of clauses 22 to 26, further comprising a pharmaceutically active agent.

[0876] 28. The lipid nanoparticle according to clause 27, wherein the pharmaceutically active agent is selected from polynucleotides, DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide, and expression vectors containing a DNA construct comprising a promoter operably linked to a sequence encoding a polynucleotide.

[0877] 29. The lipid nanoparticle according to clause 28, wherein the pharmaceutically active agent is a polynucleotide, optionally, wherein the polynucleotide is natural or artificial deoxyribonucleic acid (DNA) or ribonucleic acid (RNA);

[0878] Further optionally, wherein the polynucleotide comprises at least one chemical modification 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-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine (also known as m5U), 5-methoxyuridine, 2'-O-methyluridine, and combinations thereof. In particular, the chemical modification is N1-methylpseudouridine, 5-methoxyuridine, or a combination thereof; in particular, the chemical modification is N1-methylpseudouridine;

[0879] Further optionally, wherein the polynucleotide is ribonucleic acid (RNA);

[0880] Further optionally, wherein the RNA is selected from 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.

[0881] 30. The lipid nanoparticle according to clause 29, wherein the ratio of ionizable lipid to RNA (N / P) in the LNP is in the range of 20:1 to 2:1, particularly 10:1 to 3:1.

[0882] 31. A pharmaceutical composition comprising the lipid nanoparticle according to any one of clauses 21 to 24 and a pharmaceutically acceptable excipient or carrier.

[0883] 32. The pharmaceutical composition according to clause 31, which is a vaccine, optionally further comprising an adjuvant.

[0884] 33. A lipid nanoparticle according to any one of clauses 23 to 26 or a pharmaceutical composition according to clause 31 or 32, for use in a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the nanoparticle composition or the pharmaceutical composition;

[0885] Optionally, wherein the disease or disorder is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular and renal vascular diseases, and metabolic diseases;

[0886] Further optionally, wherein the subject is a human;

[0887] Or for use in a method of inducing an immune response in a subject, a method of therapeutic immunization for a subject, as a vaccine, or for gene therapy;

[0888] Or for the prevention or treatment of COVID-19.

[0889] 34. Use of a lipid nanoparticle according to any one of clauses 22 to 33, as an encapsulant.

[0890] Citation List

[0891] 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).

[0892] 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.

[0893] 3. US4929736.

[0894] 2. Garbiras, B.J., Marburg, S., Preparation of Carboxythiolactones and Their Active Derivatives, Synthesis, 1999, vol.2, pp.270-274; doi:10.1055 / s-1999-3377.

[0895] 3. Hassett, K.J. 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.

[0896] 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. An ionizable lipid of formula (I): or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A is selected from -NR'-C(O)-, -C(O)-NR'-, -C(O)-O- and -O-C(O)-, wherein R' is selected from H, methyl and ethyl; B is selected from -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 wherein q + t = 1, 2 or 3; R3 is a heterocycle containing at least one N atom, or R3 is: wherein Ra and Rb are independently a straight-chain or branched C1-C6 alkyl optionally substituted with a hydroxyl group; X is selected from -D-R2, wherein j, j' and j'' are independently selected from 0, 1 and 2, Z is selected from -R1 and wherein f and f' are independently selected from 0, 1, 2, 3, 4, 5 and 6, wherein g and g' are independently selected from 1, 2, 3, 4, 5 and 6, wherein D is independently selected from -C(O)-O- or -O-C(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 -OH, -COOR4 and -C(=O)SR4, wherein R4 is a C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl; provided that, - When q + t = 1, X = -D-R2, D = -O-C(O)-, A = -NH-C(O)-, B = -NH-C(O)-, Z = R1 and R1 is a straight-chain or branched alkyl group, part -(CH2) m - R1 contains at least 7 carbon atoms; and - the compound S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-homocysteine methyl ester is excluded S-(2-methoxy-2-oxoethyl)-N-pyridin-2-ylcarbonyl)-L-cysteine methyl ester and (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole.

2. The ionizable lipid according to claim 1, wherein when n = t = m = 0, q = p = 1, X is 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)-, Z is R1 and R1 is a C1-C18 alkyl, R2 and R'2 are not C7-C18 alkyl.

3. The ionizable lipid according to claim 1, which is a compound of formula (II): or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A, B, D, n, m, p, q, t, q + t, R1 and R2 are as defined in claim 1, provided that, - When q + t = 1, X = -D-R2, D = -O-C(O)-, A = -NH-C(O)-, B = -NH-C(O)-, Z = R1 and R1 is a straight-chain or branched alkyl group, part of -(CH2) m - R1 contains at least 7 carbon atoms; - the compound S-(2-methoxy-2-oxoethyl)-N-(pyridin-2-ylcarbonyl)-homocysteine methyl ester is excluded S-(2-methoxy-2-oxoethyl)-N-pyridin-2-ylcarbonyl)-L-cysteine methyl ester and (L)-CH3OOC-CH2SCH2-CH(COOCH3)-NH-CO-imidazole.

4. The ionizable lipid according to claim 3, wherein A is -NH-C(O)- or -C(O)-NH-.

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

6. The ionizable lipid according to any one of claims 3 to 5, wherein q is selected from 1 and 2, t is selected from 0 and 1, and wherein q + t = 2; or wherein q is selected from 1 and 2, t is selected from 0 and 1, q + t = 2; and R3 is a 5-membered or 6-membered ring containing one N atom and optionally a second heteroatom preferably selected from N and O; or wherein q is selected from 1 and 2, t is selected from 0 and 1, q + t = 2; and R3 is selected from the following structures:

7. The ionizable lipid according to any one of the preceding claims 3-6, 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.

8. The compound according to claim 7, wherein n is 2 or 3, m = t = 0, p = q = 2, A = B = -NH-C(O)- or -C(O)-NH-, wherein R1 is a C10-C20 branched alkyl; R2 is a C2-C30 straight-chain or branched alkyl or a C6-C24 straight-chain alkenyl or alkynyl; and R3 is: wherein Ra = Rb = C1-C4 alkyl; or wherein n = 2, R2 is a C2-C30 straight-chain or branched alkyl, and wherein R3 is: wherein Ra = Rb = C1-C4 alcohol-capped alkyl; or wherein n is 2 or 3, m = t = 0, p = q = 2, A is -NH-C(O)- or -C(O)-NH-, wherein R1 is a C10-C20 branched alkyl, R2 is a C3-C30 straight-chain or branched alkyl, and wherein R3 is an imidazole ring; or wherein n = 3, m = t = 0, p = q = 2, A is -NH-C(O)- or -C(O)-NH- wherein R1 is a C10-C20 branched alkyl, R2 is a C4-C20 straight-chain or branched alkyl, and wherein R3 is a pyrrolidine ring; or wherein n = 2, m = t = 0, p = q = 2, A is -NH-C(O)- or -C(O)-NH-, wherein R1 is a C10-C20 straight-chain or branched alkyl, R2 is a C4-C20 straight-chain or branched alkyl, and wherein R3 is N-4-methylpiperazine; or wherein n is selected from 2 or 3, m = t = 0, and p = q = 2, A = B = -NH-C(O)- or -C(O)-NH-, R1 is a C10-C22 straight-chain, branched alkyl, alkenyl or alkynyl; R2 is a C3-C30 straight-chain or branched alkyl or a C3-C30 straight-chain alkenyl or alkynyl; and wherein R3 is a morpholine ring; or wherein 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, R1 is a C10-C22 straight-chain branched alkyl; R2 is a C3-C30 straight-chain or branched alkyl or a C3-C30 straight-chain alkenyl or alkynyl, and R3 is a morpholine ring; or wherein n = q = p = 2; t = m = 0; A is B is D is R1 is a C10-C22 branched alkyl group; R2 is a C8-C20 straight-chain alkenyl or alkynyl group; or a C3-C28 straight-chain alkyl group optionally substituted with one or more substituents selected from OH, -COOR4, and -COSR4, where R4 is a C1-C6 alkyl group, a C2-C6 alkenyl group, or a C2-C6 alkynyl group; or a C12-C25 branched alkyl group; and R3 is -N(CH3)2 or 9. The ionizable lipid according to claim 1, wherein the compound is a compound of formula (III) or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A is -NH-C(O)- or -C(O)-NH-; R1, R2, and R'2 are independently selected from straight-chain or branched C1-C30 alkyl groups, C2-C30 alkenyl groups, C2-C30 alkynyl groups, wherein each of R1, R2, and R'2 is optionally substituted with one or more substituents selected from -OH, -COO(R4), and -COSR4, where R4 is a C1-C6 alkyl group, a C2-C6 alkenyl group, or a C2-C6 alkynyl group; and where B, D, n, m, p, q, t, q + t, j, j', and R3 are as defined in claim 1; or where when n = t = m = 0, q = p = 1, A = B = -NH-C(O)-, where one of j and j' is 0 and the other is 1, D = -O-C(O)-, and where when R1 is a C1-C18 alkyl group, R2 and R'2 are not C7-C18 alkyl groups.

10. The ionizable lipid according to claim 1, wherein the compound is a compound of formula (IV) or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A is -NH-C(O)- or -C(O)-NH-, and where B, D, n, m, p, q, t, q + t, f, f', g, g', and R3 are as defined in claim 1.

11. The ionizable lipid according to claim 1, which is a compound selected from the following: tridecyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0099) as defined in the specification, 8-methylnonyl 3-((4-((3-(dimethylamino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0101), tridecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0163), tetradecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0168), octadecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0169), hexadecyl 3-((3-(2-hexyldecanamido)-4-((3-morpholinopropyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0170), tridecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0178), tetradecyl 3-((3-(2-hexyldecanamido)-4-((2-(4-methylpiperazin-1-yl)ethyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0183), tridecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0194), 8-methylnonyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0196), dodecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0198), tetradecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0199), octadecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0200),Hexadecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0201), 6-Methylheptyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0202), Octyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0207), Tridecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0209), 8-Methylnonyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0211), Dodecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0213), Tetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0214), Octadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0215), Hexadecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0216), Tridecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((3-(pyrrolidin-1-yl)propyl)amino)butyl)thio)propionate (VC-LC-0241), Tridecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0256), 8-Methylnonyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0258), 8-Methylnonyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0261), Octadecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0262)Hexadecyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0263), Hexyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0268), Octyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0269), Tridecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-oleoylamido-4-oxobutyl)thio)propionate (VC-LC-0289), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butyrate (VC-LC-0294), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((8-methylnonyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0296), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0297), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0298), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-4-oxo-2-(((3-oxo-3-(tetradecyloxy)propyl)thio)methyl)butyrate (VC-LC-0299), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0300), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0301), Heptadec-9-yl 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0302)4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid nonadec-9-yl ester (VC-LC-0304), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(hexyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid nonadec-9-yl ester (VC-LC-0306), 4-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoic acid nonadec-9-yl ester (VC-LC-0307), 3-((4-((3-morpholinopropyl)amino)-3-oleoylamino-4-oxobutyl)thio)propanoic acid 2-decyltetradecyl ester (VC-LC-0353), 3-((4-((3-morpholinopropyl)amino)-3-oleoylamino-4-oxobutyl)thio)propanoic acid octadec-9-en-1-yl ester (VC-LC-0355), 3-((4-((1-methylpiperidin-4-yl)amino)-3-oleoylamino-4-oxobutyl)thio)propanoic acid 2-decyltetradecyl ester (VC-LC-0356), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0362), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-(dodecyloxy)-3-oxopropyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0366), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0367), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-(hexadecyloxy)-3-oxopropyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0369), 2-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)butanoic acid nonadec-9-yl ester (VC-LC-0370), 3-((4-((3-morpholinopropyl)amino)-4-oxo-3-stearoylaminobutyl)thio)propanoic acid 2-decyltetradecyl ester (VC-LC-0389), 6-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((6-methylheptyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-6-oxohexanoic acid 2-hexyldecyl ester (VC-LC-0418),2-decyltetradecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0428), 2-dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0429), (Z)-octadec-9-en-1-yl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0430), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyl)thio)propionate (VC-LC-0431), heptadec-9-yl 4-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0439), heptadec-9-yl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0440), heptadec-9-yl 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-oxo-3-(tridecyloxy)propyl)thio)butanoate (VC-LC-0441), heptadec-9-yl 4-((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)-2-((1-methylpiperidin-4-yl)carbamoyl)butanoate (VC-LC-0442), (Z)-heptadec-9-yl 2-((1-methylpiperidin-4-yl)carbamoyl)-4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)butanoate (VC-LC-0443), heptadec-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)propionate (VC-LC-0473), 2-hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0474), 8-methylnonyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0475), dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0477)Tetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0478), hexadecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0480), 2-decyltetradecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0487), dec-2-yn-1-yl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0488), octadec-9-en-1-yl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0489), tridecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0490), 2-hexyldecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0491), 8-methylnonyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0492), 2-decyltetradecyl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0504), dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-(((1-methylpiperidin-4-yl)methyl)amino)-4-oxobutyl)thio)propionate (VC-LC-0505), tridecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0507), 2-hexyldecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0508), 8-methylnonyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0509)2-Ethylhexyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0510), 6-Methylheptyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0515), 2-Decyltetradecyl 3-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0521), 2-Hexyldecyl 4-((1-methylpiperidin-4-yl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butyrate (VC-LC-0524), 2-Hexyldecyl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyrate (VC-LC-0525), 2-Hexyldecyl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((1-methylpiperidin-4-yl)amino)-4-oxobutyrate (VC-LC-0531), (Z)-2-Hexyldecyl 4-((1-methylpiperidin-4-yl)amino)-2-(((3-(octadec-9-en-1-yloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0539), Heptadec-9-yl 4-((3-morpholinopropyl)amino)-4-oxo-2-(((3-oxo-3-(tridecyloxy)propyl)thio)methyl)butyrate (VC-LC-0540), Heptadec-9-yl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0541), Heptadec-9-yl 2-(((3-((2-hexyldecyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0542), Heptadec-9-yl 2-(((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0543), Heptadec-9-yl 2-(((3-(dodecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutyrate (VC-LC-0544), Heptadec-9-yl 4-((3-morpholinopropyl)amino)-2-(((3-(octadecyloxy)-3-oxopropyl)thio)methyl)-4-oxobutyrate (VC-LC-0546),Heptadec-9-yl 2-(((3-(hexadecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0547), Heptadec-9-yl 2-(((3-((6-methylheptyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0548), Heptadec-9-yl 2-(((3-(docosyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0549), Heptadec-9-yl 2-(((3-(hexyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0550), Heptadec-9-yl 4-((3-morpholinopropyl)amino)-2-(((3-(octyloxy)-3-oxopropyl)thio)methyl)-4-oxobutanoate (VC-LC-0551), Heptadec-9-yl 2-(((3-((2-decyltetradecyloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0553), Heptadec-9-yl 2-(((3-(dec-2-yn-1-yloxy)-3-oxopropyl)thio)methyl)-4-((3-morpholinopropyl)amino)-4-oxobutanoate (VC-LC-0554), Tridecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0556), 2-Hexyldecyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0557), 8-Methylnonyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0558), 2-Ethylhexyl 3-((4-((1-methylpiperidin-4-yl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0559), 2-Hexyldecyl 4-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butanoate (VC-LC-0606), 2-Hexyldecyl 4-((3-((8-methylnonyloxy)-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)butyrate (VC-LC-0608), 2-Hexyldecyl 4-((3-(dodecyloxy)-3-oxopropyl)thio)-2-((3-morpholinopropyl)carbamoyl)butyrate (VC-LC-0609), 2-Hexyldecyl 2-((3-morpholinopropyl)carbamoyl)-4-((3-oxo-3-(tetradecyloxy)propyl)thio)butyrate (VC-LC-0610), 2-Hexyldecyl 3-((4-((3-(bis(2-hydroxyethyl)amino)propyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0639), 2-Hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0729), 8-Methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0730), 2-Ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0731), Dodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0732), Tetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0733), Octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0734), Hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0735), 6-Methylheptyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0736), Docosyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0737), Hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0738)Octyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0739), 2-Decyltetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0741), Dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-0742), Octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (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-triazatricosane-21-carboxylate (VC-LC-0757), Tridecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0796), 2-Hexyldecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0797), 8-Methylnonyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0798), 2-Ethylhexyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0799), Dodecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0800), Tetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0801), Octadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0802), Hexadecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0803)6-Methylheptyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0804), Docosyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0805), Hexyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0806), Octyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0807), 2-Decyltetradecyl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0809), Dec-2-yn-1-yl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0810), Octadec-9-en-1-yl 3-((3-(2-hexyldecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0811), Tridecyl 3-((4-(4-(2-hydroxyethyl)piperazin-1-yl)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0828), 8-Methylnonyl 3-((3-(2-octyldodecanamido)-4-oxo-4-((2-(piperidin-1-yl)ethyl)amino)butyl)thio)propionate (VC-LC-0848), Tridecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0864), 2-Hexyldecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0865), 8-Methylnonyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0866), 2-Ethylhexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0867), Dodecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0868)Tetradecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0869), Octadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0870), Hexadecyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0871), 6-Methylheptyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0872), Docosyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0873), Hexyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0876), Octyl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0877), Dec-2-yn-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0880), Octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0881), 6-Methylheptyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0890), Octyl 3-((4-((3-(1H-imidazol-1-yl)propyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0895), 2-Hexyldecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0919), 2-Ethylhexyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0921), Dodecyl 3-((4-((2-(diethylamino)ethyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0922)Tridecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0936), 2-Hexyldecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0937), Dodecyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0940), 6-Methylheptyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0944), Docosyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0945), Octyl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0949), (Z)-Octadec-9-en-1-yl 3-((4-((2-(dimethylamino)ethyl)(methyl)amino)-3-(2-octyldodecanamido)-4-oxobutyl)thio)propionate (VC-LC-0953), 2-Hexyldecyl 6-((4-((3-((2-hexyldecyloxy)-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)propionate (VC-LC-1065), Octyl 7-(2-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)ethyl)-2-methyl-14-((3-(octyloxy)-3-oxopropyl)thio)-6,9-dioxo-17-thia-2,5,8-triazadocos-20-oate (VC-LC-1068), Dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-1-oxo-4-((3-oxo-3-(tridecyloxy)propyl)thio)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1082)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-1083), Dioctyl 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), Dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-((2-ethylhexyl)oxy)-3-oxopropyl)thio)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1085), Dioctyl 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))dipropionate (VC-LC-1088), Dioctyl 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), 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), Dioctyl 3,3'-((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)Dioctyl 3,3'-((8-((1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((3-(deca-2-yn-1-yloxy)-3-oxopropyl)sulfanyl)-1-oxobutan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1095), Dioctyl 3,3’-((8-oxo-8-((1-oxo-4-((3-oxo-3-(tridecyloxy)propyl)sulfanyl)-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-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1097), Dioctyl 3,3’-((8-((4-((3-((2-hexyldecyl)oxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1098), Dioctyl 3,3’-((8-((4-((3-((6-methylheptyl)oxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1105), Dioctyl (Z)-3,3’-((8-((4-((3-(octadec-9-en-1-yloxy)-3-oxopropyl)sulfanyl)-1-oxo-1-((2-(piperidin-1-yl)ethyl)amino)butan-2-yl)amino)-8-oxooctane-1,3-diyl)bis(sulfanediyl))dipropionate (VC-LC-1111), (9Z,12Z)-Octadeca-9,12-dien-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)sulfanyl)propionate (VC-LC-1143), (2E,6E)-3,7,11-Trimethyldodeca-2,6,10-trien-1-yl 3-((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)sulfanyl)propionate (VC-LC-1192), Bis((E)-3,7-dimethylocta-2,6-dien-1-yl) 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)sulfanyl)methyl)succinate (VC-LC-1194)Tetradecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-1202), dodecyl 3-((4-((2-(1H-imidazol-1-yl)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-1203), bis(3,7-dimethyloctyl) 2-(((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)propionate (VC-LC-1224), (9Z,12Z)-octadeca-9,12-dien-1-yl 3-((4-((2-(bis(2-hydroxyethyl)amino)ethyl)amino)-3-(2-hexyldecanamido)-4-oxobutyl)thio)propionate (VC-LC-1230), 2-hexyldecyl 2-((2-(dimethylamino)ethyl)carbamoyl)-4-((3-((2-hexyldecyloxy)-3-oxopropyl)thio)butanoate (VC-LC-1254), bis(3,7-dimethyloctyl) 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyloxy)carbonyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1281), bis((E)-3,7-dimethyloct-2,6-dien-1-yl) 2-(((4-((2-(dimethylamino)ethyl)amino)-3-(((2-hexyldecyloxy)carbonyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1282), bis((E)-3,7-dimethyloct-2,6-dien-1-yl) 2-(((4-((2-butyloctyloxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1291), bis(3,7-dimethyloctyl) 2-(((4-((2-decyltetradecyloxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1295), bis((E)-3,7-dimethyloct-2,6-dien-1-yl) 2-(((4-((2-decyltetradecyloxy)-3-((2-(dimethylamino)ethyl)carbamoyl)-4-oxobutyl)thio)methyl)succinate (VC-LC-1296), or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them.

12. A lipid nanoparticle comprising an ionizable lipid of formula (I): or a pharmaceutically acceptable salt thereof or a stereoisomer of any of them, wherein A is selected from -NR'-C(O)-, -C(O)-NR'-, -C(O)-O-, and -O-C(O)-, where R' is selected from H, methyl, and ethyl; B is selected from -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; R3 is a heterocycle containing at least one N atom, or R3 is: where Ra and Rb are independently straight-chain or branched C1-C6 alkyl groups optionally substituted with a hydroxyl group; X is selected from -D-R2, where j, j', and j'' are independently selected from 0, 1, and 2, Z is selected from -R1 and where f and f' are independently selected from 0, 1, 2, 3, 4, 5, and 6, where g and g' are independently selected from 1, 2, 3, 4, 5, and 6, where D is independently selected from -C(O)-O- or -O-C(O)-, and wherein R1, R’1, R2, R’2 and R”2 are independently 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 -OH, -COOR4 and -C(=O)SR4, wherein R4 is C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl.

13. The lipid nanoparticle according to claim 12, further comprising: i) a non-cationic lipid; Optionally, wherein the non-cationic lipid is selected from: 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-docosanoyl-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-cholesteryl succinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16Lyso 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 16.0PE), 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 wherein the non-cationic lipid is DSPC; ii) a sterol; Optionally, wherein the sterol is selected from cholesterol, coprostanol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof; Further optionally, wherein the sterol is cholesterol; and iii) coupled lipids or PEGylated lipids; Optionally, wherein the PEGylated lipids are selected from PEGylated phosphatidylethanolamine, PEGylated phosphatidic acid, PEGylated phosphatidylcholine, PEGylated ceramide, PEGylated dialkylamine, PEGylated diacylglycerol, PEGylated dialkylglycerol, and mixtures thereof.

14. The lipid nanoparticle according to claim 13, wherein the amount of the ionizable lipid is 25 to 60 mol%, the amount of the PEGylated lipid or the coupled lipid is 0.1 to 10 mol%; the amount of the non-cationic lipid is 10 to 45 mol%; and the amount of the sterol is 10 to 40 mol%; Or wherein the amount of the ionizable lipid is 25 to 64 mol%, the amount of the PEGylated lipid or the coupled lipid is 0.1 to 1.5 mol%; and the amount of the sterol is 35 to 74 mol%.

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

16. The lipid nanoparticle according to claim 15, wherein the pharmaceutically active agent is selected from polynucleotides, DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide, and expression vectors containing DNA constructs comprising a promoter operably linked to a sequence encoding a polynucleotide.

17. The lipid nanoparticle according to claim 16, wherein the polynucleotide is natural or artificial deoxyribonucleic acid (DNA) or natural or artificial ribonucleic acid (RNA); Further optionally, wherein the polynucleotide comprises at least one chemical modification selected from 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-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine (also known as m5U), 5-methoxyuridine, 2'-O-methyluridine, and combinations thereof; in particular, the chemical modification is N1-methylpseudouridine, 5-methoxyuridine, or a combination thereof; in particular, the chemical modification is N1-methylpseudouridine; Further optionally, wherein the polynucleotide is ribonucleic acid (RNA); Further optionally, wherein the RNA is selected from 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), small hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof; in particular, the RNA is mRNA.

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

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

20. Use of the lipid nanoparticles according to any one of claims 12 - 19 as an encapsulant.

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