Silicon-containing spleen-targeted LNP delivery systems

Abstract

The present invention belongs to the field of biomedicine and drug delivery. The invention relates to LipexSil® lipid-containing nanoparticle ("LNP") compositions, that permit preferential targeting of spleen versus liver, where preferential targeting is defined as a luminescence intensity ratio of spleen and liver greater than 9 after administration of a luminescent cargo, when the different weights of spleen and liver are taken into account. In the LNP composition, a silicon-containing ionizable lipid is combined with a certain structural lipid, a helper lipid, a shield lipid and a fifth lipid component. The invention describes the production and characterization of the lipid nanoparticles and in vivo experiments demonstrating that the corresponding formulations with LipexSil® lipids as described in WO2024 / 023174 are superior to the current approach, delivering their cargo (e.g. RNA, DNA, mRNA, microRNA, siRNA, ceDNA, pDNA, circular DNA, small biologically active molecules) preferentially to the spleen.

Description

[0001] 120754P1420PC September 23, 2025

[0002] AldexChem Kft.

[0003] 1

[0004] SILICON-CONTAINING SPLEEN-TARGETED LNP DELIVERY SYSTEMS

[0005] TECHNICAL FIELD

[0006] The present invention belongs to the field of biomedicine and drug delivery.

[0007] The invention relates to LipexSil® lipid-containing nanoparticle (“LNP”) compositions, that permit preferential targeting of spleen versus liver, where preferential targeting is defined as a luminescence intensity ratio of spleen and liver greater than 9 after administration of a luminescent cargo, when the different weights of spleen and liver are taken into account.

[0008] In the LNP composition, a silicon-containing ionizable lipid is combined with a certain mixture of: a helper lipid, a structural lipid, a shield lipid and a fifth lipid component. The invention describes the production and characterization of the lipid nanoparticles and in vivo experiments demonstrating that the corresponding formulations with LipexSil® lipids as described in WO2024 / 023174 are superior to the current approach, delivering their cargo (e.g. RNA, DNA, mRNA, microRNA, siRNA, ceDNA, pDNA, circular DNA, small biologically active molecules) preferentially to the spleen.

[0009] BACKGROUND

[0010] Organ targeting is a crucial aspect of drug delivery systems, aimed at directing therapeutic agents to specific organs, thereby enhancing treatment efficacy while minimizing systemic side effects. This approach leverages the unique physiological and pathological characteristics of target organs to achieve precise delivery. Recent advancements have expanded the scope of organ targeting, incorporating sophisticated strategies like ligand-receptor interactions, nanoparticle engineering, and biocompatible materials to improve targeting specificity and efficiency.

[0011] The Selective Organ Targeting (SORT) approach discovered by Siegwart et al. (Nat. Nanotechnol. 2020, 15, 313-320) is an innovative strategy designed to optimize the distribution of nanoparticles across various organs. SORT utilizes a combination of surface chemistry, particle charge, size, and shape to direct nanoparticles to specific organs. In this approach, charge interactions can finely regulate mRNA delivery to target specific organs. For example, positively charged lipid molecules can be added to LNP formulations to specifically deliver RNA therapeutics to the lung, while negatively charged components can enable RNA delivery to the spleen. This 120754P1420PC September 23, 2025

[0012] AldexChem Kft.

[0013] 2 method significantly improves the precision of drug delivery systems, ensuring that therapeutic agents reach their intended targets with minimal distribution to non-target tissues.

[0014] Recently, several research groups have developed an array of lipids, lipoids, lipid-polymer hybrid nanoparticles, systematically varying the lipid compositions, LNP’s surface charge etc., showed enhanced spleen targeting due to preferential uptake by spleen (Adv. Mater. 2024, 36, 2311283, Angew. Chem. Int. Ed. 2023, 62, e202310395, Nano Today 2023, 52, 101943, Nat. Chem. 2024, Nat. Commun. 2024, 15, 5659, Theranostics 2024, 14(1), 1-16). The targeted delivery of therapeutic agents to the spleen holds promise for various medical applications, including hematological disorders, infectious diseases, and cancer therapy. Nanoparticles delivering drugs or gene therapies directly to the spleen can achieve higher local concentrations, improving therapeutic outcomes (Nano Today 2023, 52, 101943, Pharmaceuticals (Basel). 2022, 15(8), S)Y1 , EMBO Mol Med. 2023, 15(10): el6836, Angew. Chem. Int. Ed. 2023, 62, e202310395.

[0015] Mitchell at al. developed lipoid-containing LNPs showing a significant spleen targeting by replacing cholesterol with bile acid (Theranostics 2024, 14(1), 1-16). However, the spleen-to-liver ratio was 4 (about 36, when the value is normalized by average liver / spleen weight ratio) in case of IV, and about 9 (about 81, when the value is normalized by average liver / spleen weight ratio) in case of IP administration. In another research work, DLin-MC3-DMA and AID-lipid (amidine- incorporated degradable lipid) were combined in a certain LNP composition leading to spleen targeting with a spleen-to-liver ratio of about 5 (Nat. Chem. 2024, https: / / doi.org / 10.1038 / s41557- 024-01557-2).

[0016] Harashima et al. reached spleen-selective enhanced gene expression without selective spleen targeting of plasmid DNA, where gene expression in the spleen was about 1000 times higher than in the liver (J Control Release, 2021, 330, 753-764).

[0017] Uchida et al investigated organ-selective targeting of an mRNA polyplex with polycation and PEG achieving excellent spleen selectivity (Small Sci, 2024, 4, 2300258).

[0018] Recently, several patent applications have been published on spleen targeting with nanoparticles, but have not presented a remarkable spleen selectivity over the liver (W02020 / 051220, WO2023 / 064599, WO2023 / 129915, WO 2023 / 161378, WO2023 / 215796, W02024 / 064800, WO2024 / 125469, WO2023 / 056418, CN1679965, CN113908288, CN114712343, CN114848831, CN114887070, CN114887071, CN117088825, CN117466777, CN116392580, CN116763757, WO2023179463) 120754P1420PC September 23, 2025

[0019] AldexChem Kft.

[0020] 3

[0021] The field of organ targeting, particularly spleen targeting, continues to evolve with ongoing research focusing on enhancing the specificity and efficacy of nanoparticle-based delivery systems. Future directions include developing multimodal nanoparticles that combine multiple targeting ligands or therapeutic agents to simultaneously address different aspects of a disease or target multiple cell types within the spleen. Advanced imaging techniques will be utilized to track the biodistribution and therapeutic effects of nanoparticles in real-time, providing valuable insights into the dynamics of spleen targeting. Bridging the gap between preclinical research and clinical application by conducting rigorous clinical trials to validate the safety and efficacy of spleen- targeted therapies will also be crucial.

[0022] Although a wide range of chemically distinct lipids and nanoparticle formulations have been investigated for spleen delivery, their biodistribution outcomes are highly variable and remain difficult to predict. Even closely related lipids can result in markedly different organ targeting profiles, with minor structural or compositional changes often shifting delivery away from the spleen. This is well illustrated for example by experiments modifying the lipid tail length (Hashiba et al., Nano Lett. 2024, 24, 41, 12758-12767). Thus, despite all efforts in this field there is still a large unmet need for the identification of lipid nanoparticle-based compositions that permit the selective transfer of cargo molecules, such as mRNAs, to the spleen in a reliable manner.

[0023] SUMMARY OF THE INVENTION

[0024] Surprisingly, the present inventors have found that certain silicon-containing ionizable lipids as described in W02024 / 023174 can be combined with a certain mixture of: a helper lipid, a structural lipid, a shield lipid and a fifth lipid component result in the preferential targeting of the spleen, while structurally closely related lipids did not work. As shown in the Examples, while Lipid 1 achieves effective spleen targeting, its close analogues Lipid 6, Lipid 7, and Lipid 8 do not, despite only minor structural modifications. Conversely, Lipid 2, which is more structurally different compared to Lipid 1, nevertheless exhibited a moderate level of spleen targeting. Furthermore, Lipid 1 in formulation LNP 6 did not show spleen targeting when cholic acid was absent, underscoring the critical role of this helper lipid in enabling spleen delivery. In addition, two reference ionizable lipids commonly used in clinically validated formulations, SM-102 and ALC- 3015, which are structurally unrelated to the present lipids, were tested in analogous formulations; however, neither provided spleen targeting activity. This further underscores the inherent unpredictability of structure-activity relationships in this field and highlights that reliable spleen 120754P1420PC September 23, 2025

[0025] AldexChem Kft.

[0026] 4 targeting cannot be achieved by routine lipid modification or empirical screening alone. The present LNP systems therefore provide a non-obvious and inventive solution, delivering an unexpected technical effect of reproducible spleen selectivity beyond what has been achieved in the prior art.

[0027] Thus, in first aspect the present invention relates to lipid nanoparticle composition comprising:

[0028] (a) an ionizable cationic lipid of formula (I); or a salt thereof, wherein:

[0029] G1is unsubstituted C2-C9 alkylene;

[0030] T1is wherein b1is a bond to G1,

[0031] Xi and X2are the same or different and each independently represents O or S;

[0032] Ri is linear C1-C17 alkyl, non-linear C3-C17 alkyl, C3-C17 alkenyl containing one double bond with the proviso that there is at least one -CH2- group between the double bond and X2;

[0033] R2is linear C1-C17 alkyl, non-linear C3-C17 alkyl, C3-C17 alkenyl containing one double bond with the proviso that there is at least one -CH2- group between the double bond and the carbon linking Xi and X2; 120754P1420PC September 23, 2025

[0034] AldexChem Kft.

[0035] 5 wherein b2is a link to X

[0036] Y 1 is -O- or -CH2- or -O-CH2-CH2- wherein -CH2- is attached to Si, each X3is independently selected from the group consisting of C1-C4 alkyl or C1-C4 alkoxy,

[0037] R4 is linear C2-Cn alkyl, or non-linear C3-C17 alkyl, Cs-C2o alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and Y 1;

[0038] D1is selected from the group consisting of wherein b4is a bond to nitrogen,

[0039] Rio and Rn are independently selected from H and Ci-Cg alkyl, p is an integer selected from 2 to 6, q is an integer selected from 0 to 6,

[0040] Cyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; or

[0041] Cyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from O, N or S, wherein the heterocyclic ring is optionally substituted by RI2, wherein

[0042] RI2represents Ci-Cg alkyl;

[0043] P is selected from

[0044] (1) G2-T2wherein 120754P1420PC September 23, 2025

[0045] AldexChem Kft.

[0046] 6

[0047] G2is defined as G1above, where G1and G2may be identical or different;

[0048] T2is defined as T1above, b1is a bond to G2, and wherein T1and T2may be identical or different; or

[0049] (n) G2-T3wherein

[0050] G2is as defined above;

[0051] T3is wherein b5is a bond to G2,

[0052] X4and X5can be the same or different and each independently is O, S;

[0053] R13 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CEE- group between the double bond and X5;

[0054] R14 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CE - group between the double bond and the carbon linking X4and Xs;

[0055] (b) a structural lipid as second component selected from l,2-distearoyl-sn-glycero-3- phosphocholine (DSPC), and l,2-dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE);

[0056] (c) a helper lipid as third component selected from cholesterol, 0-sitosterol, ergosterol, campesterol, stigmasterol, and pharmaceutically acceptable salts thereof, and mixtures thereof;

[0057] (d) a shield lipid as fourth component selected from poly(ethylene glycol)-modified diacylglycerols (PEG-DAG) such as DSPE-PEG, DMG-PEG; and

[0058] (e) a fifth component which is either (el) a bile-acid component selected from cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid, ursodeoxycholic acid, and pharmaceutically acceptable salts thereof, and mixtures thereof, or 120754P1420PC September 23, 2025

[0059] AldexChem Kft.

[0060] 7

[0061] (e2) an anionic structural lipid component selected from 1,2-dioleoyl-sn-gly cero-3 - phospho-rac-(l -glycerol) (DOPG), dipalmitoylphosphatidylglycerol (DPPG), 1,2- distearoyl-sn-glycero-3 -phosphoglycerol (DSPG), 1 ,2-dimyristoyl-sn-glycero-3 - phosphoglycerol (DMPG), 1 -palmitoyl-2-oleoyl-sn-gly cero-3 -phosphoglycerol (POPG), phosphatidylserine (PS; including dioleoyl PS (DOPS) and 18:0 PS), 1,2- dipalmitoyl-sn-gly cero-3 -phosphate (DPP A), 1 ,2-dioleoyl-sn-gly cero-3 -phosphate (DOPA), and pharmaceutically acceptable salts thereof.

[0062] In a second aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue of a subject in need thereof.

[0063] In a third aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue of a subject, wherein said fifth component is cholic acid or DSPG.

[0064] In a fourth aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue of a subject, wherein the nucleic acid is selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof.

[0065] In a fifth aspect, the present invention relates to a lipid nanoparticle composition according to the invention for the treatment of a disease or condition selected from infectious diseases involving the spleen, hematologic malignancies involving the spleen, lymphoma involving the spleen, splenomegaly, functional asplenia, hyposplenism, hypersplenism, immune-related disorders.

[0066] In a sixth aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue for the treatment of a disease or condition selected from infectious diseases involving the spleen, hematologic malignancies involving the spleen, lymphoma involving the spleen, splenomegaly, functional asplenia, hyposplenism, hypersplenism, immune-related disorders, wherein the nucleic acid is selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof.

[0067] In a sixth aspect, the present invention relates to a pharmaceutical composition comprising a lipid nanoparticle composition according to the present invention comprising one or more therapeutically 120754P1420PC September 23, 2025

[0068] AldexChem Kft.

[0069] 8 active nucleic acids selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof and a pharmaceutically acceptable carrier.

[0070] BRIEF DESCRIPTION OF THE DRAWINGS

[0071] Figure 1 shows the means of luminescence signal intensity (total flux [p / s]) of the spleens and livers of each mouse group after RO administrated LNP (readout after 24 h post administration)

[0072] Figure 2 shows the means of spleen-to-liver luminescence intensity (total flux [p / s]) ratio, normalized to the respective organ weights

[0073] Figure 3 shows pictures and luminescence signal intensities (total flux [p / s]) from the spleens of individual mice (LNP 1 and LNP 3) after RO administrated LNP (readout after 24 h post administration)

[0074] DETAILED DESCRIPTION

[0075] The present invention is based on the surprising finding that novel lipid nanoparticle (LNP) compositions based on a particular class of ionizable cationic lipids could be found that preferentially target the spleen over the liver.

[0076] Thus, in a first aspect the present invention relates to a lipid nanoparticle composition, comprising an ionizable cationic lipid according to formula (I) or a salt thereof, wherein:

[0077] G1is unsubstituted C2-C9 alkylene;

[0078] T1is 120754P1420PC September 23, 2025

[0079] AldexChem Kft.

[0080] 9 wherein b1is a bond to G1,

[0081] Xi and X2are the same or different and each independently represents O or S;

[0082] Ri is linear C1-C17 alkyl, non-linear C3-C17 alkyl, C3-C17 alkenyl containing one double bond with the proviso that there is at least one -CH2- group between the double bond and X2;

[0083] R2is linear C1-C17 alkyl, non-linear C3-C17 alkyl, C3-C17 alkenyl containing one double bond with the proviso that there is at least one -CH2- group between the double bond and the carbon linking Xi and X2;

[0084] R3 is wherein b2is a link to X

[0085] Y 1 is -O- or -CH2- or -O-CH2-CH2- wherein -CH2- is attached to Si, each X3is independently selected from the group consisting of C1-C4 alkyl or C1-C4 alkoxy,

[0086] R4 is linear C2-Cn alkyl, or non-linear C3-C17 alkyl, Cs-C2o alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and Y 1;

[0087] D1is selected from the group consisting of wherein 120754P1420PC September 23, 2025

[0088] AldexChem Kft.

[0089] 10 b4is a bond to nitrogen,

[0090] Rio and Rn are independently selected from H and Ci-Cg alkyl, p is an integer selected from 2 to 6, q is an integer selected from 0 to 6,

[0091] Cyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; or

[0092] Cyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from O, N or S, wherein the heterocyclic ring is optionally substituted by R , wherein

[0093] Rn represents Ci-Cg alkyl;

[0094] P is selected from

[0095] (i) G2-T2wherein

[0096] G2is defined as G1above, where G1and G2may be identical or different;

[0097] T2is defined as T1above, b1is a bond to G2, and wherein T1and T2may be identical or different; or

[0098] (n) G2-T3wherein

[0099] G2is as defined above;

[0100] T3is wherein b5is a bond to G2,

[0101] X4and X5can be the same or different and each independently is O, S; 120754P1420PC September 23, 2025

[0102] AldexChem Kft.

[0103] 11

[0104] Ri3 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and X5;

[0105] R14 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and the carbon linking X4 and Xs;a structural lipid as second component, wherein said structural lipid is selected from 1,2- distearoyl-sn-glycero-3 -phosphocholine (DSPC), l,2-dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE), a helper lipid as third component, wherein said helper lipid is selected from cholesterol, P-sitosterol, ergosterol, campesterol, stigmasterol, and pharmaceutically acceptable salts thereof, and mixtures thereof , a shield lipid as fourth component, wherein said shield lipid is selected from poly(ethylene glycol)-modified diacylglycerols (PEG-DAG) and a fifth lipid component, wherein said fifth lipid component is either a bile-acid component selected from cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid, ursodeoxycholic acid, and pharmaceutically acceptable salts thereof, and mixtures thereof, or an anionic structural lipid component selected from l,2-dioleoyl-sn-glycero-3-phospho-rac-(l -glycerol) (DOPG), dipalmitoylphosphatidylglycerol (DPPG), l,2-distearoyl-sn-glycero-3 -phosphoglycerol (DSPG), 1 ,2-dimyristoyl-sn-gly cero-3 -phosphoglycerol (DMPG), 1 -palmitoyl-2-oleoyl-sn-gly cero-3 - phosphoglycerol (POPG), phosphatidylserine (PS; including dioleoyl PS (DOPS) and 18:0 PS), 1,2- dipalmitoyl-sn-gly cero-3 -phosphate (DPP A), 1,2-dioleoyl-sn-gly cero-3 -phosphate (DOPA), and pharmaceutically acceptable salts thereof.

[0106] In the context of the present invention, by unsubstituted C2-C9 alkylene we mean ethylene (-(CH2)2-), propylene (-(CH2)3-), butylene (-(CH2)4-), pentylene (-(CH2)5-), hexylene (-(CH2)6-), heptylene (-(CEE)?-), octylene (-(CH2)s-), or nonylene (-(CH2)9-).

[0107] By linear C1-C17 alkyl we mean methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl tridecyl, tetradecyl, pentadecyl, hexadecyl, or heptadecyl.

[0108] By non-linear C3-C17 alkyl we mean a saturated branched alkyl group containing 3 to 17 carbon atoms, e.g. isopropyl, isobutyl, 3 -methylpentyl, 2-propylpentyl, 2-methylhexyl, 2- ethylhexyl, 2-ethyldecyl, 2-propyldecyl, 2-butyldecyl, 2-pentyldecyl, 2-hexyldecyl, or 2- heptyldecyl. 120754P1420PC September 23, 2025

[0109] AldexChem Kft.

[0110] 12

[0111] By Ci-Cg alkyl group we mean a linear or non-linear alkyl group containing 1 to 6 carbon atoms, e.g. methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, tert-butyl, neopentyl, isopentyl, neohexyl or isohexyl, preferably methyl or ethyl.

[0112] By C1-C4 alkyl group we mean a linear or non-linear alkyl group containing 1 to 4 carbon atoms, preferably methyl.

[0113] By C3-C17 alkenyl containing one double bond or C3-C20 alkenyl containing one double bond we mean a linear or non-linear alkyl group as defined above containing 3 to 17 or 3 to 20 carbon atoms respectively, which contains one double bond in the chain, e.g. hex-3 -en-l-yl, oct-3 -en-l-yl, dec-3 -en-l-yl, dec-2-en-l-yl, undec-3-en-2-yl, preferably hex-3 -en-l-yl, oct-3 -en-l-yl.

[0114] By Ci-Cg alkoxy we mean -O-Ci-Cg alkyl group wherein the Ci-Cg alkyl group is as defined above.

[0115] By C1-C4 alkoxy we mean -O-C1-C4 alkyl group wherein the C1-C4 alkyl group is as defined above, like methoxy, ethoxy, propoxy, butoxy, isopropoxy, scc-butoxy or tert-butoxy.

[0116] By C3-C6 cycloalkyl we mean, for example, cyclopropyl, cyclopentyl, or cyclohexyl.

[0117] By 4 or 5 or 6 or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from N, O or S we mean an aromatic ring or an unsaturated, partly saturated or fully saturated heterocyclic ring, e.g. azetine, azetidine, pyrrolidine, oxazolidine, piperidine, piperazine, morpholine, pyrrole, imidazole, pyrazole, 1,2, 3 -triazole, 1,2,4-triazole, tetrazole, pyridine, pyrimidine, pyridazine, pyrazine, 1 ,2,4-triazine, 1,3,5-triazine, 1,2, 4, 5 tetrazine, oxazole, isoxazole, thiazole, isothiazole, 1,2-oxazine, 1,3-oxazine, 1,4-oxazine, 1 ,2-thiazine, 1,3-thiazine, 1 ,4-thiazine, azepane, azepine, 1,2-diazepane, 1,3 -diazepane, 1,4-diazepane, 1,2-diazepine, 1,3 -diazepine, 1,4- diazepine, 1,2-oxazepane, 1,3-oxazepane, 1,4-oxazepane, 1,2-oxazepine, 1,3-oxazepine, 1,4- oxazepine, 1,2-thiazepane, 1,3-thiazepane, 1,4-thiazepane, 1,2-thiazepine, 1, 3 -thi azepine, or 1,4- thiazepine ring, preferably pyrrolidine, imidazole, 1,2, 3 -triazol, piperidine, piperazine or morpholine.

[0118] By salts of the compounds of formula (I) we mean salts of the compounds of formula (I) with inorganic or organic acids. Preferred salts are those with pharmaceutically acceptable acids. The salts are e.g. chloride, sulfate, phosphate, formate, acetate, fumarate, maleate, oxalate, citrate or tartrate. The salts formed during purification or isolation are also subject of the invention. 120754P1420PC September 23, 2025

[0119] AldexChem Kft.

[0120] 13

[0121] By stereoisomers we mean optical and geometric isomers. The compounds of formula (I) may contain one or more asymmetric carbon atoms thus they can exist in the form of optical isomers, enantiomers or diastereomers. The compounds of formula (I) may contain double bounds and the groups attached to the double bond can have different (cis or trans) confirmations, e.g. cis or trans fatty acid moieties.

[0122] By anionic structural lipid we mean a phospholipid (e.g., a glycerophospholipid) that bears a net negative charge at the pH ~5-8 by virtue of an anionic headgroup (e.g., phosphoglycerol, phosphoserine, or phosphomonoester) The anionic structural lipid component selected from 1,2- dioleoyl-sn-glycero-3-phospho-rac-(l -glycerol) (DOPG), dipalmitoylphosphatidylglycerol (DPPG), l,2-distearoyl-sn-glycero-3 -phosphoglycerol (DSPG), l,2-dimyristoyl-sn-glycero-3- phosphoglycerol (DMPG), l-palmitoyl-2-oleoyl-sn-glycero-3 -phosphoglycerol (POPG), phosphatidylserine (PS; including dioleoyl PS (DOPS) and 18:0 PS), 1 ,2-dipalmitoyl-sn-glycero- 3 -phosphate (DPP A), l,2-dioleoyl-sn-glycero-3 -phosphate (DOPA) and pharmaceutically acceptable salts thereof, and mixtures thereof.

[0123] By bile acid we mean cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid, ursodeoxycholic acid and pharmaceutically acceptable salts thereof, and mixtures thereof.

[0124] In a particular embodiment, the present invention relates to lipid nanoparticle composition of claim 1, which comprises an ionizable cationic lipid according to formula (I) or a salt thereof, wherein:

[0125] G1is unsubstituted C2-C9 alkylene;

[0126] T1is 120754P1420PC September 23, 2025

[0127] AldexChem Kft. wherein b1is a bond to G1,

[0128] Xi and X2are the same or different and each independently represents O or S;

[0129] Ri is linear C1-C17 alkyl, non-linear C3-C17 alkyl, C3-C17 alkenyl containing one double bond with the proviso that there is at least one -CH2- group between the double bond and X2;

[0130] R2is linear C1-C17 alkyl, non-linear C3-C17 alkyl, C3-C17 alkenyl containing one double bond with the proviso that there is at least one -CH2- group between the double bond and the carbon linking Xi and X2;

[0131] R3 is wherein b2is a link to X

[0132] Y 1 is -CH2- or -O-CH2-CH2- wherein -CH2- is attached to Si, each X3is independently selected from the group consisting of C1-C4 alkyl or C1-C4 alkoxy,

[0133] R4 is linear C2-Cn alkyl, or non-linear C3-C17 alkyl, Cs-C2o alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and Y 1;

[0134] D1is selected from the group consisting of wherein b4is a bond to nitrogen,

[0135] Rio and Rn are independently selected from H and Ci-Cg alkyl, 120754P1420PC September 23, 2025

[0136] AldexChem Kft. p is an integer selected from 2 to 6, q is an integer selected from 0 to 6,

[0137] Cyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; or

[0138] Cyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from O, N or S, wherein the heterocyclic ring is optionally substituted by Rn, wherein

[0139] R12 represents Ci-Cg alkyl; provided that D1is not

[0140] P is selected from

[0141] (i) G2-T2wherein

[0142] G2is defined as G1above, where G1and G2may be identical or different;

[0143] T2is defined as T1above, b1is a bond to G2, and wherein T1and T2may be identical or different; or

[0144] (11) G2-T3wherein

[0145] G2is as defined above;

[0146] T3is wherein b5is a bond to G2,

[0147] X4and X5can be the same or different and each independently is O, S; 120754P1420PC September 23, 2025

[0148] AldexChem Kft.

[0149] 16

[0150] R13 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and X5;

[0151] R14 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and the carbon linking X4 and Xs;

[0152] In certain embodiments, the lipid nanoparticle composition comprises a bile-acid component as a fifth component.

[0153] In a particular embodiment, the lipid nanoparticle composition comprises cholic acid component as a fifth component.

[0154] In certain embodiments, the lipid nanoparticle composition comprises an anionic structural lipid component as a fifth component.

[0155] In a particular embodiment, the lipid nanoparticle composition comprises DSPG component as a fifth component.

[0156] In some embodiments, the lipid nanoparticle composition comprising the components are present at the following amounts, expressed as mole percent of total lipid: ionizable cationic lipid 20-70 mol %, structural lipid 2-40 mol %, helper lipid 5-40 mol %, shield lipid 0,5-10 mol %, and fifth component 1-25 mol %; wherein the total of components (ionizable cationic lipid, stuctural lipid, helper lipid shield lipid, fifth component) equals 100 mol% (excluding solvent, buffer, counterions, and payload).

[0157] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, P-sitosterol as a helper lipid and cholic acid as a fifth component.

[0158] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, cholesterol as a helper lipid and DSPG as a fifth component.

[0159] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, P-sitosterol as a helper lipid, and cholic acid as a fifth component is present at 5-20 mol % of total lipid.

[0160] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, cholesterol as a helper lipid, and DSPG as a fifth component is present at 1-8 mol % of total lipid. 120754P1420PC September 23, 2025

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[0163] In certain embodiments, the shield lipid is PEG-DSPE or PEG-DMG.

[0164] In some embodiments the shield lipid is a PEG lipid comprising a PEG moiety from 1,000 to 20,000 daltons.

[0165] In certain embodiments, the LNP has a mean hydrodynamic diameter 100-350 nm, preferably 140-300 nm.

[0166] In certain embodiments, the LNP exhibits a zeta potential of between -7.0 mV and -40.0 mV at physiological pH

[0167] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, P-sitosterol as a helper lipid, and cholic acid as a fifth component is present at 5-20 mol % of total lipid, wherein the LNP exhibits a zeta potential of between -7.0 mV and -40.0 mV at physiological pH.

[0168] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, cholesterol as a helper lipid, and DSPG as a fifth component is present at 1-8 mol % of total lipid, wherein the LNP exhibits a zeta potential of between -7.0 mV and -40.0 mV at physiological pH.

[0169] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, P-sitosterol as a helper lipid, and cholic acid as a fifth component is present at 5-20 mol % of total lipid, wherein the LNP has a mean hydrodynamic diameter 100-350 nm, preferably 140-300 nm.

[0170] In a particular embodiment, the lipid nanoparticle composition comprising DSPC as a structural lipid, cholesterol as a helper lipid, and DSPG as a fifth component is present at 1-8 mol % of total lipid, wherein the LNP has a mean hydrodynamic diameter 100-350 nm, preferably MO- SOO nm.

[0171] In a particular embodiment, the lipid nanoparticle composition comprising, an ionizable lipid, a structural lipid, a helper lipid, a shield lipid and a fifth lipid component, wherein the components are present in the following amounts expressed as mole percent of total lipid: ionizable lipid 50 mol %, DSPC 10 mol %, P-sitosterol 28,5 mol %, PEG-DSPE 1,5 mol %, and cholic acid 10 mol %. 120754P1420PC September 23, 2025

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[0174] In a particular embodiment, the lipid nanoparticle composition comprising, an ionizable lipid, a structural lipid, a helper lipid, a shield lipid and a fifth lipid component, wherein the components are present in the following amounts expressed as mole percent of total lipid: ionizable cationic lipid 50 mol %, DSPC 8 mol %, cholesterol 38,5 mol %, PEG-DMG 1,5 mol %, and DSPG 2 mol %.

[0175] In a particular embodiment, the T'-G1moiety of said ionizable cationic lipid has one of the following structures:

[0176] In a particular embodiment, the P moiety of said ionizable cationic lipid is T2-G2having one of the following structures:

[0177] In another particular embodiment, the P moiety of said ionizable cationic lipid is T3-G2moiety having one of the following structures:

[0178] In some embodiments, the D1moiety of said ionizable cationic lipid has one of the following structures: 120754P1420PC September 23, 2025

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[0181] In a particular embodiment, the D1moiety of said ionizable cationic lipid has one of the following structures:

[0182] In a particular embodiment, the ionizable cationic lipid is chosen from the following structures:

[0183] In a particular embodiment, the LNP permits preferential targeting of spleen versus liver. Preferential targeting is defined as a ratio of normalized luminescence intensities, wherein the normalized luminescence intensity of each organ is defined as the luminescence intensity divided by the organ weight, such that preferential targeting corresponds to a value ( / Spieen / »ispieen) / ( iiver / »2iiver) is greater than 10. Herein, I represents the total flux [p / s] of the corresponding organ and m represents the weight [mg] of the corresponding organ 120754P1420PC September 23, 2025

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[0185] 20

[0186] In particular embodiments, the LNP has a composition selected from LNP 1, LNP 2, LNP 3, LNP 4 and LNP 5 (see Table 3).

[0187] In a particular embodiment, the LNP permits preferential targeting of spleen versus liver, where preferential targeting is defined as a normalized luminescence intensity ratio of spleen and liver greater than 50.

[0188] In particular embodiments, the LNP has a composition selected from LNP 1, LNP 2, LNP 3 and LNP 4 (see Table 3).

[0189] In a particular embodiment, the LNP permits preferential targeting of spleen versus liver, where preferential targeting is defined as a normalized luminescence intensity ratio of spleen and liver greater than 100.

[0190] In a particular embodiment, the LNP permits preferential targeting of spleen versus liver, where preferential targeting is defined as a normalized luminescence intensity ratio of spleen and liver greater than 200.

[0191] In a particular such embodiments, the LNP has the composition of LNP 1 (see Table 3).

[0192] In some embodiments, the LNP is loaded with at least one payload selected from the group consisting of mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, small biologically active molecule and any combination thereof.

[0193] In some embodiments, the lipid nanoparticle composition is used for delivering one or more nucleic acids to spleen tissue of a subject in need thereof.

[0194] In some embodiments, the lipid nanoparticle composition is used for delivering one or more nucleic acids to spleen tissue of a subject in need thereof, wherein the fifth component cholic acid or DSPG.

[0195] In some embodiments, the lipid nanoparticle composition for use in delivering one or more nucleic acids to spleen tissue of a subject, wherein the nucleic acid is selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof.

[0196] In some embodiments, the lipid nanoparticle composition is used for delivering one or more nucleic acids to spleen tissue for the treatment of a disease or condition selected from infectious diseases involving the spleen, hematologic malignancies involving the spleen, lymphoma involving 120754P1420PC September 23, 2025

[0197] AldexChem Kft.

[0198] 21 the spleen, splenomegaly, functional asplenia, hyposplenism, hypersplenism, or immune-related disorders.

[0199] In a second aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue of a subject in need thereof.

[0200] In a third aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue of a subject, wherein said fifth component is cholic acid or DSPG.

[0201] In a fourth aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue of a subject, wherein the nucleic acid is selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof.

[0202] In a fifth aspect, the present invention relates to a lipid nanoparticle composition according to the invention for the treatment of a disease or condition selected from infectious diseases involving the spleen, hematologic malignancies involving the spleen, lymphoma involving the spleen, splenomegaly, functional asplenia, hyposplenism, hypersplenism, immune-related disorders.

[0203] In a sixth aspect, the present invention relates to a lipid nanoparticle composition according to the invention for use in delivering one or more nucleic acids to spleen tissue for the treatment of a disease or condition selected from infectious diseases involving the spleen, hematologic malignancies involving the spleen, lymphoma involving the spleen, splenomegaly, functional asplenia, hyposplenism, hypersplenism, immune-related disorders, wherein the nucleic acid is selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof.

[0204] In a sixth aspect, the present invention relates to a pharmaceutical composition comprising a lipid nanoparticle composition according to the present invention comprising one or more therapeutically active nucleic acids selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof and a pharmaceutically acceptable carrier. 120754P1420PC September 23, 2025

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[0207] EXAMPLES

[0208] The various embodiments of this application can be more clearly understood through the following illustrative examples. These examples are provided for explanatory purposes and do not restrict the scope of the application. The synthetic routes of the Examples (Lipid 1, Lipid 2, Lipid 3, Lipid 4, Lipid 5, Lipid 6, Lipid 7, Lipid 8) are based on methodologies, as further disclosed in WO2024 / 023174, and can be routinely performed by a person skilled in the art without undue experimentation.

[0209] Lipid 6, Lipid 7, and Lipid 8 did not exhibit significant spleen targeting in vivo. These results highlight the unexpected and non-obvious nature of the present lipid systems, as even close analogues fail to achieve the technical effect observed with Lipid 1.

[0210] IN VITRO TOXICITY STUDY ON HEPG2 CELL

[0211] HepG2 cell line (human hepatocellular carcinoma) were maintained in DMEM medium supplemented with 10% PBS. Cell toxicity measurements were carried out in 384-well microtiter plates with 1000 cells seeded in each well. Test measurements were carried out in triplicate, and control measurements in quadruplicate. After an overnight incubation, the cells were treated with an increasing concentration of the test compounds (6.25, 12.5, 25, 50, 100, 200, 400, 800 pM). Solvent (ethanol) content of wells treated with the lipids was 6,4% and with the reference lipids (SM-102 and ALC-0315) was 16% at the highest applied dose of 800 pM. 48 h post-treatment, following visual inspection under an inverted microscope, cell viability was measured by resazurin assay. Viability of treated cells was calculated relative to that of ethanol vehicle controls. Average values and SD were calculated from repeated measurements. CRCs were obtained and corresponding IC50 values were calculated where applicable.

[0212] Resazurin reagent:

[0213] Resazurin reagent (Sigma- Aldrich) was dissolved in PBS (pH 7.4) at 0.15 mg / ml concentration, 0.22 pm filtered and aliquoted at -20 °C. 10 pl resazurin stock solution was added to wells. After 5 hours incubation at 37 °C under 5 % CO2 fluorescence (530 nm excitation / 580 nm emission) was recorded on a multimode microplate reader.

[0214] Toxicology results were comparable across the lipids, and no major discrepancies were identified that would explain the differential spleen targeting.

[0215] Table 1: Determined IC50 values 120754P1420PC September 23, 2025

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[0218] LNP FORMULATION AND CHARACTERIZATION

[0219] Formulation

[0220] The lipid solutions used contained four or five components: an ionizable lipid, DSPC, 0- sitosterol, DSPE-PEG2000 and cholic acid in a mol ratio of 50: 10:28.5: 1.5:10, an ionizable lipid, DSPC, Cholesterol, DMG-PEG2000, DSPGin a molar ratio of 50:8:38.5:1.5:2 or an ionizable lipid, DSPC, 0-sitosterol andDSPE-PEG2000 in a molar ratio of 50:10:38.5:1.5. All lipids were dissolved 120754P1420PC September 23, 2025

[0221] AldexChem Kft.

[0222] 24 in 100% ethanol in a concentration ranging from 5-50 mM and stored at -20°C. Before formulating, lipids were heated to 37°C vortexed for 2 minutes and cooled to room temperature. The mRNA (in case of LNP 1, LNP 2, LNP 6 RiboPro Off-The-Shelf Flue mRNA with Capl while in the case of LNP 3, 4, 5, 7, 8, 9, 10, 11, 12, 13 Trilink CleanCap® Flue mRNA (NIMePsU) were used) was diluted with 100 mM citrate (pH 4.0) and RNAse free water to a final concentration 0.15 pg / pL in 10 m citrate. The lipid and mRNA solutions were mixed in a 3 : 1 (mRNA aqueous: lipid in EtOH) ratio by microfluidic mixing with a total flow-rate of 12 ml / min (total flow rate (TFR)) using formulation machine and microfluidic cartridge. Next the samples were transferred to 100 kDa MWCO, SpectraPor Biotech dialysis tubing, and closed with designated clamps. Particles were buffer exchanged to PBS (pH 7.4) by dialysis for two times two hours and 1 time overnight. In the last dialysis step, penicillin-streptomycin (final concentration of 100 units / mL of penicillin and 100 pg / mL of streptomycin) were added to the PBS buffer. Formulations were placed onto Ultra Centrifugal Filters (100 kDa MWCO, Amicon) and concentrated to 100 ng / pL in the final buffer PBS (pH 7.4).

[0223] DLS (Size, PDI and Zeta potential)

[0224] Characterization of mRNA-LNPs was done with dynamic light scattering (DLS, Zetasizer Pro ZSU3205, Malvern Panalytical) to determine size, poly dispersity and zeta potential of the LNP samples. Therefore, all dialyzed sample were diluted 70 times in lx PBS buffer, pH7.4, and size and PDI were measured by using the “Size & Zeta” option of the “ZS Explorer” software. Size and PDI were measured after formulation, dialysis and concentration of the samples. To measure the zeta potential samples were 10 x diluted in 0.1 x PBS buffer, pH 7.4 and pipetted into a Folded Capillary Zeta Cell (Malvern Panalytical, cat number DTS1070 ). Zeta potential was measured by using the “Size & Zeta” option of the “ZS Explorer” software.

[0225] Encapsulation assay

[0226] Free and total mRNA concentrations were determined by the RiboGreen assay (Invitrogen, cat. number 15797239) according to the manufacturer’s protocol. In short, the dialysed samples were lOOx diluted in TE buffer, pH 7.4. Then, 50 pL of all samples were mixed 1 : 1 with TE buffer, or with TE buffer containing 1% Triton X-100 buffer. Next samples were incubated in a 96 well plate at 37° C for 10 min to lyse mRNA-LNP in the presence of Triton X-100. Meanwhile, the RiboGreen reagent was diluted 1 : 100 in TE buffer, pH 7.5. After 10 min the plate with the samples was cooled to room temperature and 100 pL of RiboGreen solution were added to each well. The fluorescent signal in all wells was determined using a plate reader (excitation = 480 nm, emission 120754P1420PC September 23, 2025

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[0229] = 525 nm). A standard curve (0.1-1.0 ng / l RNA) was used to calculate the total and free concentration of mRNA in the LNP samples.

[0230] Table 2: Physico-chemical parameters of produced LNPs 120754P1420PC September 23, 2025

[0231] AldexChem Kft.

[0232] IN VIVO TRANSFECTION STUDY ON MOUSE

[0233] 24 h before administration, animals were moved from the animal house to treatment area and were assigned to treatment groups (n=3). Animals receiving the same treatment were kept in the same cage. Measurements of body weight were performed before and 24 hours after LNP administration. Following administration animals were monitored regularly for any adverse clinical observations. Animals were dosed intravenously (retro orbital vein, RO) with 5 pg Flue mRNA- loaded LNPs in 100 pl. After treatment with LNP formulations (see Table 2) animals were kept in the treatment area for 24 hours. After 24 hours all animals received 90 mg / kg D-luciferin (15 mg / ml; 6 pl / g of body mass) intraperitoneally. 15 min later they were sacrificed by cervical dislocation and dissected. Selected organs (liver, lung, kidney, spleen) were collected and placed in 6 well plates and imaged by the IVIS In Vivo Imaging System. The results are shown in Table 3.

[0234] CONCLUSION

[0235] The present invention describes lipid nanoparticles that demonstrate significant spleentargeting capabilities. The primary component of these LNPs is the proprietary LipexSil® ionizable lipid, which is combined with an optimal ratio of the said further lipid components in an appropriate formulation, exhibit enhanced spleen-targeting properties. This has been validated through the transfection of Flue mRNA, where it was observed that spleen targeting is substantially more pronounced compared to the liver. These lipids are relatively simple from synthetic point of view, and they can be produced from readily available raw materials. Furthermore, these ionizable lipids show lower toxicity on HepG2 cell line relative to the reference lipids used. Based on these findings, these formulations represent promising candidates for further clinical developments programs. 120754P1420PC September 23, 2025

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[0238] Table 3: Luminescence signal in LNP treated animals 120754P1420PC September 23, 2025

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[0240] 28 average weight of spleen (24 mice): 113,09 mg, average weight of liver (24 mice): 1014,75 mg, weight ratio: 8,97, and bdl: below detection limit, nd: no data

Claims

39AMENDED CLAIMS received by the International Bureau on 09 March 2026 (09.03.2026)1. A lipid nanoparticle composition comprising:(a) an ionizable cationic lipid of formula (I);or a salt thereof, wherein:G1is unsubstituted C2-C9 alkylene;T1iswherein b1is a bond to G1,Xi and X2are the same or different and each independently represents O or S;R-2 is linear C1-C17 alkyl, non-linear C3-C17 alkyl, C3-C17 alkenyl containing one double bond with the proviso that there is at least one -CH2- group between the double bond and the carbon linking Xi and X2;R3 iswherein b2is a link to Xi,40Y i is -CH2- or -O-CH2-CH2- wherein -CH2- is attached to Si, each X3is independently selected from the group consisting of C1-C4 alkyl or C1-C4 alkoxy,R4 is linear C2-C17 alkyl, or non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and Y 1;D1is selected from the group consisting ofwherein b4is a bond to nitrogen,Rio and Rn are independently selected from H and Ci-Ce alkyl, p is an integer selected from 2 to 6, q is an integer selected from 0 to 6,Cyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from N, O or S; orCyi is a 4-, 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 heteroatoms selected from O, N or S, wherein the heterocyclic ring is optionally substituted by R12, whereinR12 represents Ci-Cs alkyl; provided that D1is notP is selected from(1) G2-T2whereinG2is defined as G1above, where G1and G2may be identical or different;41T2is defined as T1above, b1is a bond to G2, and wherein T1and T2may be identical or different; or(11) G2-T3whereinG2is as defined above;T3is whereinb5is a bond to G2,X4and X5can be the same or different and each independently is O, S;Ri3 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and X5;R14 is linear C2-C17 alkyl, non-linear C3-C17 alkyl, C3-C20 alkenyl containing one double bond with the proviso there is at least one -CH2- group between the double bond and the carbon linking X4and X5;(b) a structural lipid as second component selected from l,2-distearoyl-sn-glycero-3- phosphocholine (DSPC), and l,2-dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE);(c) a helper lipid as third component selected from cholesterol, P-sitosterol, ergosterol, campesterol, stigmasterol, and pharmaceutically acceptable salts thereof, and mixtures thereof;(d) a shield lipid as fourth component selected from poly(ethylene glycol)-modified diacylglycerols (PEG-DAG) such as DSPE-PEG, DMG-PEG; and(e) a fifth component which i IsS either(el) a bile-acid component selected from cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid, ursodeoxycholic acid, and pharmaceutically acceptable salts thereof, and mixtures thereof, or(e2) an anionic structural lipid component selected from l,2-dioleoyl-sn-glycero-3-42 phospho-rac-(l -glycerol) (DOPG), dipalmitoylphosphatidylglycerol (DPPG), 1,2- distearoyl-sn-glycero-3 -phosphoglycerol (DSPG), l,2-dimyristoyl-sn-glycero-3- phosphoglycerol (DMPG), l-palmitoyl-2-oleoyl-sn-glycero-3 -phosphoglycerol (POPG), phosphatidylserine (PS; including dioleoyl PS (DOPS) and 18:0 PS), 1,2- dipalmitoyl-sn-glycero-3 -phosphate (DPP A), 1 ,2-dioleoyl-sn-gly cero-3 -phosphate (DOPA), and pharmaceutically acceptable salts thereof.

2. The lipid nanoparticle composition of claim 1, wherein said fifth component is a bile-acid component.

3. The lipid nanoparticle composition of claim 2, wherein said fifth component is cholic acid.

4. The lipid nanoparticle composition of claim 1, wherein said fifth component is an anionic structural lipid component.

5. The lipid nanoparticle composition of claim 4, wherein said fifth component is DSPG.

6. The lipid nanoparticle composition of any one of claims 1 to 5, wherein said components are present at the following amounts, expressed as mole percent of total lipid: ionizable cationic lipid 20-70 mol %, structural lipid 2-40 mol %, helper lipid 5-40 mol %, shield lipid 0,5-10 mol %, and fifth component 1-25 mol %; wherein the total of components (ionizable cationic lipid, structural lipid, helper lipid shield lipid, fifth component) equals 100 mol% (excluding solvent, buffer, counterions, and payload).

7. The lipid nanoparticle composition of any one of claims 1 to 3 and 6, wherein said structural lipid is DSPC, said helper lipid is 13-sitosterol and said fifth component is cholic acid.

8. The lipid nanoparticle composition of claim 7, wherein said fifth component is present at 5- 20 mol % of total lipid.

9. The lipid nanoparticle composition any one of claims 1 and 4 to 6, wherein said structural lipid is DSPC, said helper lipid is cholesterol and said fifth component is DSPG.

10. The lipid nanoparticle composition of claim 9, wherein said fifth component is present at 1- 8 mol % of total lipid.

11. The lipid nanoparticle composition of any one of claims 1 to 10, wherein said shield lipid is PEG-DSPE or PEG-DMG.

12. The lipid nanoparticle composition of any one of claims 1 to 11, wherein the T^G1moiety of said ionizable cationic lipid has one of the following structures:4313. The lipid nanoparticle composition of any one of claims 1 to 12, wherein the P moiety of said ionizable cationic lipid is T2-G2having one of the following structures:

14. The lipid nanoparticle composition of any one of claims 1 to 12, wherein the P moiety of said ionizable cationic lipid is a T3-G2moiety, wherein G2is as defined above, and T3has the following structure:

15. The lipid nanoparticle composition of any one of claims 1 to 14, wherein the D1moiety of said ionizable cationic lipid has one of the following structures:

16. The lipid nanoparticle composition of claim 15, wherein the D1moiety of said ionizable cationic lipid has one of the following structures:

17. The lipid nanoparticle composition of any one of claims 1 to 16, wherein said ionizable cationic lipid has one of the following structures:4418. The lipid nanoparticle composition according to claim 17, wherein the following components are present in the following amounts expressed as mole percent of total lipid: ionizable cationic lipid 50 mol %, DSPC 10 mol %, 13-sitosterol 28,5 mol %, PEG-DSPE 1,5 mol %, and cholic acid 10 mol %.

19. The lipid nanoparticle composition according to claim 17, wherein the following components are present in the following amounts expressed as mole percent of total lipid: ionizable cationic lipid 50 mol %, DSPC 8 mol %, cholesterol 38,5 mol %, PEG-DMG 1,5 mol %, and DSPG 2 mol %.

20. The lipid nanoparticle composition of any one of claims 1 to 19 for use in delivering one or more nucleic acids to spleen tissue of a subject in need thereof.

21. The lipid nanoparticle composition of any one of claims 1 to 19 for use in delivering one or more nucleic acids to spleen tissue of a subject, wherein said fifth component is cholic acid or DSPG.

22. The lipid nanoparticle composition of any one of claims 1 to 19 for use in delivering one or more nucleic acids to spleen tissue of a subject, wherein the nucleic acid is selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof.

23. The lipid nanoparticle composition any one of claims 1 to 19 for use in delivering one or more nucleic acids to spleen tissue for the treatment of a disease or condition selected from infectious diseases involving the spleen, hematologic malignancies involving the spleen,45 lymphoma involving the spleen, splenomegaly, functional asplenia, hyposplenism, hypersplenism, immune-related disorders.

24. The lipid nanoparticle composition any one of claims 1 to 19 for use in delivering one or more nucleic acids to spleen tissue for the treatment of a disease or condition selected from infectious diseases involving the spleen, hematologic malignancies involving the spleen, lymphoma involving the spleen, splenomegaly, functional asplenia, hyposplenism, hypersplenism, immune-related disorders, wherein the nucleic acid is selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof.

25. A pharmaceutical composition comprising a lipid nanoparticle composition of any one of claims 1 to 19 comprising one or more therapeutically active nucleic acids selected from mRNA, cDNA, ceDNA, pDNA, ceDNA, microRNA, siRNA, gRNA, CAS9 mRNA, saRNA, circRNA, modified RNA, antisense oligonucleotide, targeted nucleic acid, or a combination thereof and a pharmaceutically acceptable carrier.

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