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US-20260124306-A1 - NUCLEIC ACID-CONTAINING LIPID NANO-PARTICLE AND USE THEREOF

US20260124306A1US 20260124306 A1US20260124306 A1US 20260124306A1US-20260124306-A1

Abstract

The present invention provides a lipid nanoparticle containing the following (a) to (c): (a) a nucleic acid encoding a chimeric antigen receptor (CAR) or an exogenous T cell receptor (TCR); (b) a cationic lipid; and (c) a non-cationic lipid. The present invention also provides a CAR- or exogenous TCR-expressing immunocyte obtained by introducing the lipid nanoparticle into in vivo or ex vivo T cells, and an in vivo or ex vivo therapeutic approach using the immunocytes for disease such as cancer and the like.

Inventors

  • Shinobu Kuwae
  • Satoru Matsumoto

Assignees

  • TAKEDA PHARMACEUTICAL COMPANY LIMITED

Dates

Publication Date
20260507
Application Date
20251229
Priority Date
20171227

Claims (11)

  1. 1 .- 34 . (canceled)
  2. 35 . A method of inducing a T cell to express a chimeric antigen receptor or an exogenous T cell receptor comprising administering to a mammal in need thereof a lipid nanoparticle comprising the following (a) to (c): (a) a nucleic acid encoding a chimeric antigen receptor or an exogenous T cell receptor; (b) a cationic lipid; and (c) a non-cationic lipid, wherein the lipid nanoparticle has a surface ligand targeting a T cell and wherein the lipid nanoparticle introduces the nucleic acid encoding the chimeric antigen receptor or the exogenous T cell receptor into an in vivo T cell of the mammal to induce an expression thereof.
  3. 36 . The method according to according to claim 35 , wherein said cationic lipid is a compound represented by the formula (I): wherein L1 is a C1-22 alkylene group, a C2-22 alkenylene group or a C3-22 alkadienylene group, n is an integer of 0 or 1, R1 is (1) a hydrogen atom, (2) a linear C1-22 alkyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group, (3) a linear C2-22 alkenyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group, or (4) a linear C3-22 alkadienyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group, R2 is —CH2-O—CO—R5, —CH2-CO—O—R5 or —R5, R3 is —CH2-O—CO—R6, —CH2-CO—O—R6 or —R6, R4 is a hydrogen atom, —CH2-O—CO—R7, —CH2-CO—O—R7 or —R7, R5, R6 and R7 are each independently (1) a linear C1-22 alkyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group, (2) a linear C2-22 alkenyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group, or (3) a linear C3-22 alkadienyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group, R8 and R9 are each independently, a C1-6 alkyl group, or a salt thereof.
  4. 37 . The method according to claim 35 , wherein the aforementioned nucleic acid is mRNA or DNA.
  5. 38 . The method according to claim 35 , wherein the aforementioned non-cationic lipid is phospholipid, cholesterol and/or PEG lipid.
  6. 39 . The method according to claim 35 , wherein the aforementioned ligand is a ligand comprising an antigen binding domain of one or more antibodies selected from the group consisting of an antibody against CD3, an antibody against CD4, an antibody against CD8 and an antibody against CD28.
  7. 40 . The method according to claim 35 , wherein the aforementioned ligand is a ligand comprising an antigen binding domain of an antibody against CD3.
  8. 41 . The method according to claim 35 , wherein the aforementioned cationic lipid is a compound 7: or a salt thereof.
  9. 42 . The method according to claim 35 , wherein the aforementioned cationic lipid is a compound 12: or a salt thereof.
  10. 43 . The method according to claim 35 , wherein the aforementioned cationic lipid is a compound 21: or a salt thereof.
  11. 44 . The method according to claim 35 , wherein the aforementioned cationic lipid is a compound 35: or a salt thereof.

Description

TECHNICAL FIELD The present invention relates to lipid nanoparticles containing a nucleic acid encoding a chimeric antigen receptor or a T cell receptor, a method for expressing a chimeric antigen receptor or an exogenous T cell receptor in a immunocyte of interest by using the lipid nanoparticles, a pharmaceutical use thereof, and the like. BACKGROUND OF THE INVENTION The research and development of cancer immunotherapy using CAR-T cells or TCR-T cells introduced with a gene of chimeric antigen receptor (CAR) or T-cell receptor (TCR) derived from cancer antigen-specific killer T cell is progressing rapidly. Current CAR-T cell therapy, such as Kymriah (trade name) and Yescarta (trade name), which were approved in the U.S., generally includes producing CAR-T cells by transfecting T cells collected from patients with CAR genes ex vivo using viral vectors such as lentiviral vector, and administering the CAR-T cells to the patients. However, this method has the problem that the production cost becomes high due to the cost of cell culture and preparation of viral vectors. If introduction of CAR or exogenous TCR selectively into immunocytes, such as T cells, in vivo is possible, ex vivo preparation is not necessary and CAR- or TCR-immunocell therapy with low production cost can be provided. In addition, if CAR or exogenous TCR can be selectively introduced into immunocytes such as T cells in ex vivo without using viral vectors requiring high production cost, the cost of virus residue testing, etc. will be eliminated, and CAR- or TCR-immunocell therapy with low production cost can be provided. The ex vivo or in vivo transfection of CAR into T cells has been reported which uses nanoparticles containing aggregates of CAR-encoding plasmid DNA and a cationic polymer that are coated with a non-cationic polymer conjugated with anti-CD3 antibody fragments (patent document 1, non-patent document 1), or nanocarrier containing mesoporous silica encapsulating CAR-encoding DNA in the pores and coated with a lipid having a surface modified with an anti-CD3 antibody (patent document 2). Apart therefrom, techniques have been reported for delivering siRNA to a target cell by encapsulating the target siRNA in “lipid nanoparticles (LNP)”, which do not have an internal pore structure and are composed of a cationic lipid, a non-cationic helper lipid, and a ligand for delivery to the target cell. For example, ex vivo or in vivo transfection of siRNA for CD45 into T cells by using an anti-CD4 antibody fragment as a targeted ligand has been reported (patent document 3, non-patent document 2). To date, however, there is no report that a nucleic acid (e.g., mRNA, DNA) encoding CAR or exogenous TCR has been selectively introduced into immunocytes such as T cells by using LNP. DOCUMENT LIST Patent Documents patent document 1: US 2017/0296676patent document 2: US 2016/0145348patent document 3: WO 2016/189532 Non-Patent Documents non-patent document 1: Nature Nanotechnology 12, 813-820 (2017)non-patent document 2: ACS Nano, 2015, 9(7), 6706-6716 SUMMARY OF INVENTION Technical Problem The purpose of the present invention is to provide a novel transfection technology capable of efficiently introducing CAR or exogenous TCR selectively into immunocytes such as T cells in vivo or ex vivo, thereby providing CAR- or TCR-immunocell therapy with low production cost. Another purpose of the present invention is to provide a safer CAR- or TCR-immunocell therapy that avoids the problem of antigenicity by viral proteins. Solution to Problem The present inventors have conducted intensive studies in an attempt to achieve the above-mentioned purposes and succeeded in efficiently introducing a nucleic acid encoding CAR or exogenous TCR selectively into immunocytes, such as T cell, in vivo and ex vivo by using LNP, which resulted in the completion of the present invention. Accordingly, the present invention provides the following. [1] A lipid nanoparticle comprising the following (a) to (c): (a) a nucleic acid encoding a chimeric antigen receptor or an exogenous T cell receptor;(b) a cationic lipid; and(c) a non-cationic lipid. [2] The lipid nanoparticle of [1], wherein the aforementioned cationic lipid is a compound represented by the formula (I): whereinL is a C1-22 alkylene group, a C2-22 alkenylene group or a C3-22 alkadienylene group,n is an integer of 0 or 1,R1 is (1) a hydrogen atom,(2) a linear C1-22 alkyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group,(3) a linear C2-22 alkenyl group optionally substituted by one or two substituents selected from a linear C1-22 alkyl group and a linear C2-22 alkenyl group, or(4) a linear C3-22 alkadienyl group optionally substituted by one or two substituents selected from a linear C22 alkyl group and a linear C2-22 alkenyl group, R2 is —CH2—O—CO—R5, —CH2—CO—O—R5 or —R5,R3 is —CH2—O—CO—R6, —CH2—CO—O—R6 or —R6,R4 is a hyd