Search

CN-122028909-A - Lipid nanoparticles with non-covalent bifunctional conjugates for active targeting

CN122028909ACN 122028909 ACN122028909 ACN 122028909ACN-122028909-A

Abstract

The nanoparticle complex comprises a bifunctional conjugate capable of non-covalent binding to a lipid nanoparticle in the nanoparticle complex.

Inventors

  • R. A. wessel heft
  • AMIT SINGH
  • LUO ZHIWEI
  • LIN FENG
  • LAN TING
  • LI SANPENG

Assignees

  • 莫特治疗股份有限公司
  • 菲鹏弘济生物(深圳)有限公司

Dates

Publication Date
20260512
Application Date
20241023
Priority Date
20231024

Claims (20)

  1. 1. A nanoparticle complex, the nanoparticle complex comprising (A) Lipid nanoparticles comprising a surface component; (b) The double-function combination is used for the combination, the bifunctional conjugate comprises (I) A self-binding moiety non-covalently bound to the surface component, (Ii) A target conjugate moiety capable of binding to a targeted cell surface component of a target cell, Wherein the self-binding moiety is linked to the target binding moiety, and (C) Cargo carried by the lipid nanoparticle.
  2. 2. The nanoparticle complex according to claim 1, wherein the cargo is an RNA selected from the group consisting of circular RNA, mRNA, non-coding RNA, dsRNA, miRNA, siRNA and tRNA, preferably the RNA is circular RNA.
  3. 3. The nanoparticle complex of claim 1 or 2, wherein the lipid nanoparticle comprises an ionizable lipid, a helper lipid, a PEG-modified lipid, and a cholesterol-based lipid, optionally the lipid nanoparticle further comprises a biotin-modified lipid, optionally the biotin-modified lipid is biotin-modified cholesterol.
  4. 4. A nanoparticle complex according to any one of claims 1 to 3, wherein the ionisable lipid is a cationic lipid ：HGT4003、HGT5000、HGT5001、HGT5002、DOTMA、DOGS、DOSPA、DOTAP、DODAP、DOTMA、DSDMA、DODMA、DLinDMA、DLenDMA、DODAC、DDAB、DMRIE、CLinDMA、CpLinDMA、DMOBA、DOcarbDAP、DLinDAP、DLincarbDAP、DLinCDAP、DLin-K-XTC2-DMA、DLin-KC2-DMA、AA3-DLinXTC、ALNY-100、NC98-5、C12-200、MC2、MC3、MC4、ALC-0315、SM-102、ATX-001、ATX-100、 lipid 2, lipid 5、9A1P9、OF-Deg-Lin、80-O16B、93-O17S、93-O17O、306-O12B、113-O16B、306Oi10、113-O12B、cKK-E12、98N12-5、OF-02、TLC053、LP01、BAMEA-O16B、CL1、BP lipid 310, lipid A9, L319 and ICE selected from the group consisting of.
  5. 5. The nanoparticle complex of any one of claims 1-4, wherein the helper lipid is a non-cationic lipid selected from DSPC, DOPC, DPPC, DOPG, DPPG, DOPE, POPC, POPE, POPG, DEPE, lecithin, DOPE-mal, DPPE, DMPE, DSPE, 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, and SOPE.
  6. 6. The nanoparticle complex of any one of claims 1-5, wherein the PEG-modified lipid is a polyethylene glycol (PEG) chain covalently attached to a lipid having an alkyl chain, wherein the PEG chain has a length of 100-20000 Da, optionally the PEG chain has a length of 100, 200, 400, 600, 800, 1000, 2000, 4000, 8000, 10000, 20000 Da, optionally the alkyl chain has a length of C 6 -C 20 .
  7. 7. The nanoparticle complex of any one of claims 1-6, wherein the PEG-modified lipid is a glycol (PEG) -modified phospholipid or ceramide, or a combination thereof.
  8. 8. The nanoparticle complex of any one of claims 1-7, wherein the PEG-modified lipid is DMG-PEG2000, DSPE-PEG2000, or a combination thereof.
  9. 9. The nanoparticle complex of any one of claims 1-8, wherein the cholesterol-based lipid is selected from cholesterol, hydroxycholesterol and derivatives thereof, DC-cholesterol HCl, HAPC-cholesterol, MHAPC-cholesterol, DMHAPC-cholesterol, DMPAC-cholesterol, GL-67, lc-10, pegylated cholesterol or derivatives thereof, DC-Choi, l, 4-bis (3-N-oleylamino-propyl) piperazine, ICE, vitamin D2, vitamin D3, calcipotriol, stigmasterol, b-sitosterol, betulin, lupeol, ursolic acid, oleanolic acid, lipid derivatives of cholesterol lipids, wherein the lipid may be myristate, oleate, nervonate or hemisuccinate.
  10. 10. The nanoparticle complex of any one of claims 1-9, wherein the lipid nanoparticle comprises (1) ALC-0315, cholesterol, DOPE and DMG-PEG2000, or (2) ALC-0315, cholesterol, DSPC, DMG-PEG2000 and DSPE-PEG2000, or (3) SM102, cholesterol, DOPE, DMG-PEG2000, DSPE-PEG2000 and 18:1PG, or (4) Lipid 5, cholesterol, DSPC, DMG-PEG2000, DSPE-PEG2000 and 18:1PG, or (5) SM102, cholesterol, DSPC and DMG-PEG2000.
  11. 11. The nanoparticle complex of any one of claims 1-10, wherein the surface component is an ionizable lipid, a helper lipid, a PEG-modified lipid, or a biotin-modified lipid, optionally the biotin-modified lipid is biotin-modified cholesterol.
  12. 12. The nanoparticle complex of any one of claims 1-11, wherein the self-conjugate moiety is a first antigen-binding fragment that specifically binds to the surface component, optionally the first antigen-binding fragment specifically binds to PEG, optionally the first antigen-binding fragment specifically binds to methoxy of PEG or the backbone of PEG, preferably the first antigen-binding fragment specifically binds to methoxy of PEG, optionally the first antigen-binding fragment specifically binds to the cholesterol-based lipid, optionally the cholesterol-based lipid is conjugated to biotin, and the first antigen-binding fragment specifically binds to the biotin.
  13. 13. The nanoparticle complex of claim 12, wherein the first antigen-binding fragment is derived from IgG, igA, igM, igE or IgD, optionally the first antigen-binding fragment is a Fab, VHH antibody, scFv, scFab or diabody.
  14. 14. The nanoparticle complex of any one of claims 1-13, wherein the target conjugate moiety is IgG, igA, igM, igE or an Fc of IgD or a fragment thereof.
  15. 15. The nanoparticle complex of any one of claims 1-14, wherein the targeted cell surface component of the target cell is a protein, glycosylated RNA, lipid, or lipid raft, optionally the protein is a receptor, optionally the receptor is an Fc receptor, fcgR1, fcgR2, FCgr, fcgR4, fceR1, fceR2, fcaR, fcuR, fcdR, or an isoform thereof, or C1qR or FcRn, optionally the receptor is selected from CD2, CD3, CD4, CD5, CD7, CD8, CD25, CD RA 45, CD45RO, CTLA4, and PL-L1.
  16. 16. The nanoparticle complex of any one of claims 1-15, wherein the target cell is an antigen presenting cell, a monocyte, a neutrophil, a macrophage, a dendritic cell, a mast cell, a T cell, a natural killer cell, a Coulopfer cell, a B cell, or a tumor cell, optionally the T cell is a CD3+ T cell, a CD4+ T cell, a CD8+ T cell, a regulatory T cell, a cytotoxic T cell, or a helper T cell.
  17. 17. The nanoparticle complex of any one of claims 1-16, wherein the target conjugate moiety is a second antigen-binding fragment that specifically binds to the receptor, optionally the second antigen-binding fragment is a Fab, VHH antibody, scFv, scFab, or diabody.
  18. 18. The nanoparticle complex of any one of claims 1-17, wherein the self-binding moiety is covalently linked to the target-binding moiety, optionally the self-binding moiety is linked to the target-binding moiety via a linker, optionally the linker comprises Fc or a fragment thereof, optionally the linker comprises a multimerization domain.
  19. 19. The nanoparticle complex of any one of claims 1-17, wherein the self-conjugate moiety is non-covalently linked to the target conjugate moiety.
  20. 20. The nanoparticle composite of any one of claims 1-19, wherein the nanoparticle composite has at least one of the following functions as compared to a reference nanoparticle composite: (a) Substantially avoiding uptake by hepatocytes, (B) Substantially contacting a cell displaying said target cell surface component of said target cell, and (C) Substantially reducing accelerated blood clearance of the nanoparticle complex, Wherein the reference nanoparticle complex is a nanoparticle complex without the bifunctional conjugate.

Description

Lipid nanoparticles with non-covalent bifunctional conjugates for active targeting Sequence listing The sequence listing, which is filed by electronic filing and incorporated herein by reference, is contained in a file named "088177-8001WO01", which is 75,282 bytes (as measured in Microsoft Windows) and created at 10, 22, 2024. Cross Reference to Related Applications The present application claims priority from U.S. provisional application 63/592,915 filed on 10/24 of 2023, the disclosure of which is incorporated herein by reference. Technical Field The present disclosure relates generally to lipid nanoparticles. In particular, the disclosure relates to lipid nanoparticles with non-covalent bifunctional conjugates for active targeting and uses thereof. Background Liposomes are synthetic lipid vesicles that can encapsulate molecules of various sizes in their interior space. Liposomes have been widely studied as carriers for delivering substances to cells and tissues in vivo. (Gregorian, liposome Technology, volumes I, II, III, CRC Press, boca Raton, 1985.) nanoparticles, like liposomes, have also been studied for use as drug carriers, especially PEG-NPs, which can be obtained by covalent attachment (PEGylation) of poly (ethylene glycol) nanoparticles and thus exhibit increased drug bioavailability, prolonged blood circulation half-life and hinder reticuloendothelial system (RES) capture. However, NPs (especially PEG-NPs) often accumulate near the tumor but do not penetrate into the tumor mass, and some drugs cannot readily diffuse from the PEG-NPs to the target cells. Although several studies provide some ideas to increase specific targeting and intracellular uptake, there are still other problems to be solved, such as manufacturability of the currently covalently functionalized targeted lipid nanoparticles, blood clearance of particles that lead to loss of efficacy in the presence of particle-bound antibodies, and targeting of particles to extrahepatic tissues and cell types, among others. Furthermore, the ability to treat only a narrow spectrum of diseases is also a pending problem. Taking the technical solution disclosed in WO 2015134411 (A1) as an example, the upgraded NPs disclosed therein can only deliver small molecules and thus only treat cancer. Accordingly, there is a need in the related art for products that can overcome the above-described drawbacks. Disclosure of Invention Throughout this disclosure, the articles "a," "an," and "the" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. For example, "an antibody" means one antibody or more than one antibody. In one aspect, the present disclosure provides a nanoparticle complex comprising (a) a Lipid Nanoparticle (LNP) comprising a surface component, (b) a bifunctional conjugate comprising (i) a self-conjugate moiety that is non-covalently bound to the surface component, (ii) a target-conjugate moiety capable of binding to a targeted cell surface component of a target cell, wherein the self-conjugate moiety is linked to the target-conjugate moiety, and (c) a cargo carried by the lipid nanoparticle, wherein the cargo is one or more nucleic acid molecules. In some embodiments, the nucleic acid is DNA or RNA. In some embodiments, the nucleic acid is selected from dsDNA, ssDNA, circular RNA, mRNA, non-coding RNA, dsRNA, miRNA, siRNA, and tRNA, preferably circular RNA, mRNA, or siRNA. In some embodiments, the lipid nanoparticle comprises an ionizable lipid, a helper lipid, a PEG-modified lipid, and a cholesterol-based lipid or other lipid with similar functionality. In some embodiments, the lipid nanoparticle comprises a biotin-modified lipid (such as biotin-modified cholesterol) or other modified lipid with similar function. In some embodiments, the ionizable lipid is a cationic lipid ：HGT4003、HGT5000、HGT5001、HGT5002、DOTMA、DOGS、DOSPA、DOTAP、DODAP、DOTMA、DSDMA、DODMA、DLinDMA、DLenDMA、DODAC、DDAB、DMRIE、CLinDMA、CpLinDMA、DMOBA、DOcarbDAP、DLinDAP、DLincarbDAP、DLinCDAP、DLin-K-XTC2-DMA、DLin-KC2-DMA、AA3-DLinXTC、ALNY-100、NC98-5、C12-200、MC2、MC3、MC4、ALC-0315、SM-102、ATX-001、ATX-100、 selected from the group consisting of lipid 2, lipid 5、9A1P9、OF-Deg-Lin、80-O16B、93-O17S、93-O17O、306-O12B、113-O16B、306Oi10、113-O12B、cKK-E12、98N12-5、OF-02、TLC053、LP01、BAMEA-O16B、CL1、BP lipid 310, lipid A9, L319, and ICE. In some embodiments, the helper lipid is a non-cationic lipid selected from DSPC, DOPC, DPPC, DOPG, DPPG, DOPE, POPC, POPE, POPG, DEPE, lecithin, DOPE-mal, DPPE, DMPE, DSPE, 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, and SOPE. In some embodiments, the PEG-modified lipid is a polyethylene glycol chain of up to 10 kDa in length covalently attached to a lipid having one or more alkyl chains of C6-C20 length. In some embodiments, the PEG-modified lipid is a glycol (PEG) -modified phospholipid or ceramide. In some embodiments, the surface component is an ionizable lipid, a helper lipid, a PEG-modified lipid,