# US 2019/0314291 A1 · Compositions and methods for delivery of agents to immune cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0314291 A1 - Document ID: US20190314291A1 - Status: Published application - Kind: A1 - Application number: 16/262,758 - Filed: 2019-01-30 - Published: 2019-10-17 - Earliest claimed: 2018-01-30 - Assignees: ModernaTX, Inc. - Applicants: ModernaTX, Inc. - Inventors: Gilles BESIN; Luis BRITO; Stephen G. HOGE; Edward HENNESSY; Mark CORNEBISE; Kerry BENENATO; Staci SABNIS; Michael W. DANNEMAN - Web page: https://patentgrep.com/patent/US20190314291A1 ## CPC - A61K 9/127: Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant - A61K 9/5123: {Organic compounds, e.g. fats, sugars} - A61K 9/5146: {obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides} - A61K 39/0011: {Cancer antigens} - A61K 39/245: Herpetoviridae, e.g. herpes simplex virus - A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells - A61K 40/31: Chimeric antigen receptors [CAR] - A61K 40/421: {Immunoglobulin superfamily} - A61K 47/24: containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids - A61K 47/28: Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid - A61P 35/00: Antineoplastic agents - C07K 14/525: Tumour necrosis factor [TNF] - C07K 14/70503: {Immunoglobulin superfamily} - C12N 15/88: using microencapsulation, e.g. using {amphiphile} liposome vesicle - A61K 2239/48: Blood cells, e.g. leukemia or lymphoma - C07K 14/7051: {T-cell receptor (TcR)-CD3 complex} - C07K 2319/03: containing a transmembrane segment - C12N 2710/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The disclosure features immune cell delivery lipid nanoparticle (LNP) compositions that allow for enhanced delivery of agents, e.g., nucleic acids, such as therapeutic and/or prophylactic RNAs, to immune cells, in particular T cells, as well as B cells, dendritic cells and monocytes. The LNPs comprise an effective amount of an immune cell delivery potentiating lipid such that delivery of an agent by an immune cell delivery LNP is enhanced as compared to an LNP lacking the immune cell delivery potentiating agent. Methods of using the immune cell delivery LNPs for delivery of agents, e.g., nucleic acid delivery, for protein expression, for modulating immune cell activity and modulating immune responses are also disclosed. ## Claims ### Claim 1 An immune cell delivery lipid nanoparticle comprising: (i) an ionizable lipid; (ii) a sterol or other structural lipid; (iii) a non-cationic helper lipid or phospholipid; (iv) an agent for delivery to an immune cell, and (v) optionally, a PEG-lipid wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to an immune cell. ### Claim 2 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises a PEG-lipid, and wherein the agent comprises an mRNA encoding a protein of interest. ### Claim 3 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid or (iii) the non-cationic helper lipid or phospholipid or (v) the PEG lipid is a C1q binding lipid that binds to C1q and/or promotes the binding of the LNP to C1q, as compared to a lipid nanoparticle lacking the C1q binding lipid. ### Claim 4 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the enhanced delivery is relative to a lipid nanoparticle lacking the immune cell delivery potentiating lipid. ### Claim 5 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the enhanced delivery is relative to a suitable control. ### Claim 6 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the agent stimulates protein expression in the immune cell. ### Claim 7 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the agent inhibits protein expression in the immune cell. ### Claim 8 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the agent encodes a soluble protein that modulates immune cell activity. ### Claim 9 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the agent encodes an intracellular protein that modulates immune cell activity. ### Claim 10 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the agent encodes a transmembrane protein that modulates immune cell activity. ### Claim 11 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the agent enhances immune function. ### Claim 12 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the agent inhibits immune function. ### Claim 13 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the immune cell is a T cell. ### Claim 14 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the immune cell is a B cell. ### Claim 15 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the immune cell is a dendritic cell or a myeloid cell. ### Claim 16 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises a phytosterol, or salt or ester thereof, or a combination of a phytosterol and cholesterol or a salt thereof. ### Claim 17 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the phytosterol is selected from the group consisting of β-sitosterol, stigmasterol, β-sitostanol, campesterol, brassicasterol, and combinations thereof. ### Claim 18 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the phytosterol comprises a sitosterol or a salt or an ester thereof. ### Claim 19 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the phytosterol comprises a stigmasterol or a salt or an ester thereof. ### Claim 20 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the phytosterol is beta-sitosterol or a salt or an ester thereof. ### Claim 21 (canceled) ### Claim 22 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the immune cell is a T cell and the phytosterol or a salt or ester thereof is selected from the group consisting of β-sitosterol, β-sitostanol, campesterol, brassicasterol, Compound S-140, Compound S-151, Compound S-156, Compound S-157, Compound S-159, Compound S-160, Compound S-164, Compound S-165, Compound S-170, Compound S-173, Compound S-175 and combinations thereof. ### Claim 23 (depends on 22) The immune cell delivery lipid nanoparticle of claim 22, wherein the phytosterol is β-sitosterol. ### Claim 24 (depends on 22) The immune cell delivery lipid nanoparticle of claim 22, wherein the phytosterol is β-sitostanol. ### Claim 25 (depends on 22) The immune cell delivery lipid nanoparticle of claim 22, wherein the phytosterol is campesterol. ### Claim 26 (depends on 22) The immune cell delivery lipid nanoparticle of claim 22, wherein the phytosterol is brassicasterol. ### Claim 27 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the immune cell is a monocyte or a myeloid cell and the phytosterol or a salt or ester thereof is selected from the group consisting of β-sitosterol, and stigmasterol, and combinations thereof. ### Claim 28 (depends on 27) The immune cell delivery lipid nanoparticle of claim 27, wherein the phytosterol is β-sitosterol. ### Claim 29 (depends on 27) The immune cell delivery lipid nanoparticle of claim 27, wherein the phytosterol is stigmasterol. ### Claim 30 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises a sterol, or a salt or ester thereof, and cholesterol, wherein the immune cell is a monocyte or a myeloid cell and the sterol or a salt or ester thereof is selected from the group consisting of β-sitosterol-d7, brassicasterol, Compound S-30, Compound S-31 and Compound S-32. ### Claim 31 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the mol % cholesterol is between about 1% and 50% of the mol % of phytosterol present in the lipid nanoparticle. ### Claim 32 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the mol % cholesterol is between about 10% and 40% of the mol % of phytosterol present in the lipid nanoparticle. ### Claim 33 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the mol % cholesterol is between about 20% and 30% of the mol % of phytosterol present in the lipid nanoparticle. ### Claim 34 (depends on 16) The immune cell delivery lipid nanoparticle of claim 16, wherein the mol % cholesterol is about 30% of the mol % of phytosterol present in the lipid nanoparticle. ### Claim 35 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the ionizable lipid comprises a compound of any of Formulae (I I), (I IA), (I IB), (I II), (I IIa), (I IIb), (I IIc), (I IId), (I IIe), (I IIf), (I IIg), (I III), (I VI), (I VI-a), (I VII), (I VIII), (I VIIa), (I VIIIa), (I VIIIb), (I VIIb-1), (I VIIb-2), (I VIIb-3), (I VIIc), (I VIId), (I VIIIc), (I VIIId), (I IX), (I IXa1), (I IXa2), (I IXa3), (I IXa4), (I IXa5), (I IXa6), (I IXa7), or (I IXa8). ### Claim 36 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the ionizable lipid comprises a compound selected from the group consisting of Compound X, Compound Y, Compound I-48, Compound I-50, Compound I-109, Compound I-111, Compound I-113, Compound I-181, Compound I-182, Compound I-244, Compound I-292, Compound I-301, Compound I-309, Compound I-317, Compound I-321, Compound I-322, Compound I-326, Compound I-328, Compound I-330, Compound I-331, Compound I-332, Compound I-347, Compound I-348, Compound I-349, Compound I-350, Compound I-352 and Compound I-M. ### Claim 37 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the ionizable lipid comprises a compound selected from the group consisting of Compound X, Compound Y, Compound I-321, Compound I-292, Compound I-326, Compound I-182, Compound I-301, Compound I-48, Compound I-50, Compound I-328, Compound I-330, Compound I-109, Compound I-111 and Compound I-181. ### Claim 38 (canceled) ### Claim 39 (depends on 37) The immune cell delivery lipid nanoparticle of claim 37, wherein immune cell is a T cell and the ionizable lipid comprises a compound selected from the group consisting of Compound I-301, Compound I-321, and Compound I-326. ### Claim 40 (depends on 36) The immune cell delivery lipid nanoparticle of claim 36, wherein immune cell is a monocyte or a myeloid cell and the ionizable lipid comprises a compound selected from the group consisting of Compound X, Compound I-109, Compound I-111, Compound I-181, Compound I-182, and Compound I-244. ### Claim 41 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, wherein the non-cationic helper lipid or phospholipid comprises a compound selected from the group consisting of DSPC, DPPC, DMPC, DMPE, DOPC, Compound H-409, Compound H-418, Compound H-420, Compound H-421 and Compound H-422. ### Claim 42 (depends on 41) The immune cell delivery lipid nanoparticle of claim 41, wherein the phospholipid is DSPC. ### Claim 43 (depends on 41) The immune cell delivery lipid nanoparticle of claim 41, wherein the immune cell is a T cell and the non-cationic helper lipid or phospholipid comprises a compound selected from the group consisting of DSPC, DMPE, and Compound H-409. ### Claim at position 44 -46. (canceled) ### Claim 47 (depends on 41) The immune cell delivery lipid nanoparticle of claim 41, wherein the immune cell is a monocyte or a myeloid cell and the non-cationic helper lipid or phospholipid comprises a compound selected from the group consisting of DOPC, DMPE, and Compound H-409. ### Claim at position 46 -50. (canceled) ### Claim 51 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises a PEG-lipid. ### Claim 52 (depends on 51) The immune cell delivery lipid nanoparticle of claim 51, wherein the PEG-lipid is selected from the group consisting of a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixtures thereof. ### Claim 53 (depends on 51) The immune cell delivery lipid nanoparticle of claim 51, wherein the PEG lipid comprises a compound selected from the group consisting of Compound P-415, Compound P-416, Compound P-417, Compound P-419, Compound P-420, Compound P-423, Compound P-424, Compound P-428, Compound P-L1, Compound P-L2, Compound P-L3, Compound P-L4, Compound P-L6, Compound P-L8, Compound P-L9, Compound P-L16, Compound P-L17, Compound P-L18, Compound P-L19, Compound P-L22, Compound P-L23 and Compound P-L25. ### Claim 54 (canceled) ### Claim 55 (depends on 51) The immune cell delivery lipid nanoparticle of claim 51, wherein the PEG lipid comprises a compound selected from the group consisting of Compound P-428, Compound PL-16, Compound PL-17, Compound PL-18, Compound PL-19, Compound PL-1, and Compound PL-2. ### Claim 56 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises about 30 mol % to about 60 mol % ionizable lipid, about 0 mol % to about 30 mol % non-cationic helper lipid or phospholipid, about 18.5 mol % to about 48.5 mol % sterol or other structural lipid, and about 0 mol % to about 10 mol % PEG lipid. ### Claim 57 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises about 35 mol % to about 55 mol % ionizable lipid, about 5 mol % to about 25 mol % non-cationic helper lipid or phospholipid, about 30 mol % to about 40 mol % sterol or other structural lipid, and about 0 mol % to about 10 mol % PEG lipid. ### Claim 58 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises about 50 mol % ionizable lipid, about 10 mol % non-cationic helper lipid or phospholipid, about 38.5 mol % sterol or other structural lipid, and about 1.5 mol % PEG lipid. ### Claim 59 (depends on 56) The immune cell delivery lipid nanoparticle of claim 56, wherein the mol % sterol or other structural lipid is 18.5% phytosterol and the total mol % structural lipid is 38.5%. ### Claim 60 (depends on 56) The immune cell delivery lipid nanoparticle of claim 56, wherein the mol % sterol or other structural lipid is 28.5% phytosterol and the total mol % structural lipid is 38.5%. ### Claim 61 (canceled) ### Claim 62 (depends on 1) The immune cell delivery lipid nanoparticle of claim 1, which comprises: (i) about 50 mol % ionizable lipid, wherein the ionizable lipid is a compound selected from the group consisting of Compound I-301, Compound I-321, and Compound I-326; (ii) about 10 mol % phospholipid, wherein the phospholipid is DSPC; (iii) about 38.5 mol % structural lipid, wherein the structural lipid is selected from β-sitosterol and cholesterol; and (iv) about 1.5 mol % PEG lipid, wherein the PEG lipid is Compound P-428. ### Claim 63 A method of delivering an agent to an immune cell, the method comprising contacting the immune cell with an immune cell delivery lipid nanoparticle comprising: (i) an ionizable lipid; (ii) a sterol or other structural lipid; (iii) a non-cationic helper lipid or phospholipid; (iv an agent for delivery to an immune cell, and (v) optionally, a PEG-lipid wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to an immune cell, such that the agent is delivered to the immune cell. ### Claim 64 (depends on 63) The method of claim 63, wherein the agent comprises a nucleic acid encoding the protein of interest, such that expression of the protein of interest is induced in or on the immune cell. ### Claim 65 (depends on 63) The method of claim 63, comprising contacting a T cell with the immune cell delivery lipid nanoparticle, wherein the agent comprises a nucleic acid that modulates T cell activation or activity, such that T cell activation or activity is modulated. ### Claim 66 A method of increasing an immune response to a protein, the method comprising contacting immune cells with an immune cell delivery lipid nanoparticle comprising: (i) an ionizable lipid; (ii) a sterol or other structural lipid; (iii) a non-cationic helper lipid or phospholipid; (iv) an agent comprising a nucleic acid molecule, and (v) optionally, a PEG-lipid wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to immune cells, such that the immune response to the protein is increased. ### Claim 67 A method of increasing a T cell response to a cancer antigen, the method comprising contacting the T cell with an immune cell delivery lipid nanoparticle comprising: (i) an ionizable lipid; (ii) a sterol or other structural lipid; (iii) a non-cationic helper lipid or phospholipid; (iv) an agent comprising an mRNA encoding a chimeric antigen receptor (CAR) that binds the cancer antigen, and (v) optionally, a PEG-lipid wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to immune cells, such that the T cell response to the cancer antigen is increased. ### Claim 68 (depends on 66) The method of claim 66, wherein the agent comprises an mRNA encoding an antigen of interest, such that the immune response to the antigen of interest is enhanced in the subject, as compared to the immune response to the antigen of interest induced by an LNP encapsulating the mRNA encoding the antigen of interest but lacking the immune cell delivery potentiating lipid. ### Claim 69 (depends on 63) The method of claim 63, comprising contacting a B cell with the immune cell delivery lipid nanoparticle, wherein the agent comprises a nucleic acid molecule that modulates B cell activation or activity, such that B cell activation or activity is modulated. ### Claim at position 66 -128. (canceled)