EP-4734953-A1 - LIPID NANOPARTICLE FORMULATIONS FOR CELL THERAPY AND RELATED METHODS

Abstract

Provided is a lipid formulation which is substantially free from cholesterol and which is mixed with nucleic acid to form lipid nucleic acid nanoparticles which are capable of transfecting cells of the immune system in a less destructive manner than electroporation.

Inventors

• HARVIE, PIERROT
• JEFFS, Lloyd Brian
• ZHANG, Ruo Yu
• KAZEMIAN, Mohammadreza
• CHAKRAPANI, Harish
• JAIN, NIKITA
• THOMAS, Anitha
• KONDRATOWICZ, Andrew

Assignees

• Global Life Sciences Solutions Canada ULC

Dates

Publication Date

20260506

Application Date

20231222

Claims (20)

1. 1. A lipid formulation for forming a lipid-based nanoparticle suitable for transfecting nucleic acid cargo into cells of hematopoietic lineage comprising an ionizable lipid, a phospholipid, and optionally a stabilizer.
2. 2. A lipid formulation comprising an ionizable lipid, a phospholipid, and optionally a stabilizer, for forming a lipid-based nanoparticle suitable for transfecting a nucleic acid cargo into T cells, Jurkat cells, hematopoietic stem cells, natural killer cells, or antigen presenting cells.
3. 3. The lipid formulation of claim 1, further comprising a nucleic acid cargo.
4. 4. The lipid formulation of claim 2, further comprising a nucleic acid cargo.
5. 5. The lipid formulation of claim 4, wherein the nucleic acid cargo comprises chimeric antigen receptor (CAR) encoded mRNA.
6. 6. The lipid formulation of claim 5, wherein the transfected cells are T cells.
7. 7. The lipid formulation of claim 5, wherein the transfected cells are hematopoietic stem cells.
8. 8. The lipid formulation of claim 4, wherein the nucleic acid cargo comprises a gene editing nuclease and a guide RNA to perform permanent gene knockout and/or insertion.
9. 9. The lipid formulation of claim 8, wherein the transfected cells are T cells.
10. 10. The lipid formulation of claim 8, wherein the transfected cells are hematopoietic stem cells.
11. 11. The lipid formulation of claim 3, wherein the nucleic acid cargo comprises an mRNA encoding a protein to correct a genetic deficiency.
12. 12. The lipid formulation of claim 3, wherein the nucleic acid cargo comprises an mRNA encoding a protein to treat disease.
13. 13. The lipid formulation of any of claims 1-12, wherein the stabilizer comprises from about 0.1 to about 10.0 %Mol.
14. 14. The lipid formulation of claim 13, wherein the stabilizer is a PEGylated lipid.
15. 15. The lipid formulation of claim 13, wherein the stabilizer is a polysarcosine.
16. 16. The lipid formulation of claim 13, wherein the stabilizer is vitamin E polyethylene glycol succinate (TPGS).
17. 17. The lipid formulation of any of claims 1-12, wherein the stabilizer is absent.
18. 18. The lipid formulation of any one of claims 1-17, wherein the ionizable lipid comprises from about 10 to about 50 %Mol.
19. 19. The lipid formulation of any one of claims 1-18 wherein the ionizable lipid has a jasmonic acid headgroup.
20. 20. The lipid formulation of any one of claims 1-18 wherein the ionizable lipid has a tetrahydrofuran head group.

Description

LIPID NANOPARTICLE FORMULATIONS FOR CELL THERAPY AND RELATED METHODS CROSS-REFERENCE TO RELATED APPLICATIONS [001] This patent application claims priority to International Application No. PCT/US2023/069303, filed June 28, 2023, which claims the benefit of U.S. Provisional Application 63/357,094, filed June 30, 2022, which are incorporated by reference in their entireties herein. BACKGROUND [002] Nucleic-acid-based cell therapy reagents offer advantages over electroporation and traditional oncological treatments in terms of safety and efficacy. In particular, RNA cell therapies are subject to degradation by exonucleases and endonucleases in vivo without a delivery system, so they need a carrier. [003] Lipid nanoparticles (also referred to herein as “LNP”) generally consist of different lipids, each serving distinct functions. LNP can have a lipidic or aqueous core and may contain bilayer structures depending on the abundance of each type of lipids. [004] Generally, the components of LNP formulations include: ionizable cationic lipids, which spontaneously encapsulate negatively-charged nucleic acids by a combination of attractive electrostatic interactions with and hydrophobic interactions; neutral phospholipids to reduce charge-related toxicity and to maintain structure of the LNP; and cholesterol to stabilize the LNP and help with cell entry, and a lipid-conjugated polyethylene glycol (PEG). [005] Recognized challenges to creating successful LNP cell therapy are safety, manufacturability, stability, and efficacy. There is a continued need in the art to provide additional solutions to a better cell therapy delivery agent with both high efficiency and high live cell yield. The present disclosure provides for ameliorating at least some of the disadvantages of the prior art. These and other advantages of the present disclosure will be apparent from the description as set forth below. [006] It will be appreciated that this background description has been created to aid the reader and is not to be taken as an indication that any of the indicated problems were themselves appreciated in the art. While the described principles can, in some aspects and embodiments, alleviate the problems inherent in other systems, it will be appreciated that the scope of the protected innovation is defined by the attached claims and not by the ability of any disclosed feature to solve any specific problem noted herein. BRIEF SUMMARY [007] The disclosure provides, e.g., lipid formulations and related methods suitable for forming nucleic acid-based cell therapy reagents in lipid nanoparticles. In an aspect, the disclosure provides a lipid formulation for forming a lipid-based nanoparticle suitable for transfecting nucleic acid cargo into cells of hematopoietic lineage comprising an ionizable lipid, a phospholipid, and optionally a stabilizer. [008] In another aspect, the disclosure provides a lipid formulation comprising an ionizable lipid, a phospholipid, and optionally a stabilizer, for forming a lipid-based nanoparticle suitable for transfecting a nucleic acid cargo into T cells, Jurkat cells, hematopoietic stem cells, natural killer cells, or antigen presenting cells. [009] In an aspect, the disclosure provides a lipid formulation for forming a lipid-based nanoparticle suitable for transfecting nucleic acid cargo into cells of hematopoietic lineage comprising an ionizable lipid, a phospholipid, and optionally a stabilizer, wherein the formulation is substantially free of cholesterol. [0010] In another aspect, the disclosure provides a lipid formulation comprising an ionizable lipid, a phospholipid, and optionally a stabilizer, for forming a lipid-based nanoparticle suitable for transfecting a nucleic acid cargo into T cells, Jurkat cells, hematopoietic stem cells, natural killer cells, or antigen presenting cells, wherein the formulation is substantially free of cholesterol. [O H ] In a further aspect, the disclosure provides a method for screening lipid composition of lipid nanoparticles for preferential delivery of RNA to T cells, comprising: (a) preparing a plurality of lipid nanoparticles with different lipid compositions and a reporter RNA, (b) administering the lipid nanoparticles to Jurkat cells, (c) measuring the relative abundance of reporter RNA, (d) comparing the relative abundance of reporter RNA to a threshold, and (e) identifying lipid compositions above threshold as candidates for preferential delivery of RNA to T cells. [0012] Further and alternative aspects and features of the disclosed principles will be appreciated from the following detailed description and the accompanying drawings. Other aspects and features of the present invention will become apparent to those ordinarily skilled in the art upon review of the following description of specific embodiments of the invention in conjunction with the accompanying figures. Accordingly, it is to be understood that both the foregoing general description and the fo