EXTRACELLULAR VESICLE CONJUGATES AND USES THEREOF
The present disclosure relates to extracellular vesicles (e.g., exosomes) comprising a biologically active molecule covalently linked to the extracellular vesicle via a maleimide moiety, which may be useful as an agent for the prophylaxis or treatment of cancer and other diseases. Also provided herein are methods for producing the extracellular vesicles and methods for using the extracellular vesicles to treat diseases or disorders.
1 - 84 . (canceled)
85 . An extracellular vesicle (EV) comprising a biologically active molecule and a cleavable linker.
86 . The cleavable linker of claim 85 , wherein the cleavable linker comprises an enzymatic-cleavable linker.
87 . The cleavable linker of claim 86 , wherein the enzymatic-cleavable linker comprises a reductase-cleavable linker, a protease cleavable-linker, a nuclease-cleavable linker, an esterase-cleavable linker, a phosphatase-cleavable linker, an amidase-cleavable linker, and/or an oxidase-cleavable linker.
88 . The cleavable linker of claim 85 , wherein the cleavable linker comprises a redox cleavable linker.
89 . The cleavable linker of claim 88 , wherein the redox cleavable linker comprises a disulfide bond.
90 . The cleavable linker of claim 85 , wherein the cleavable linker comprise a protease-cleavable linker.
91 . The EV of claim 90 , wherein the protease-cleavable linker comprises valine, alanine, citrulline, phenylalanine, N-methyl-valine, lysine, cyclohexyl-alanine, beta-alanine, glycine, or any combination thereof.
92 . The EV of claim 85 , wherein the cleavable linker further comprises a self-immolative spacer.
93 . The EV of claim 85 , wherein the cleavable linker further comprises a non-cleavable-linker.
94 . The EV of claim 93 , wherein the non-cleavable linker comprises tetraethylene glycol (TEG), polyethylene glycol (PEG), hexaethylene glycol (HEG), succinimide, a glycine spacer, a C2 to C12 alkyl spacer, or any combination thereof.
95 . The EV of claim 94 , wherein the glycine spacer is glycine, or glycine-glycine.
96 . The EV of claim 85 , wherein the cleavable linker is attached to a scaffold protein or to a scaffold lipid.
97 . The EV of claim 96 , wherein the scaffold protein comprises a prostaglandin F2 receptor negative regulator (PTGFRN) polypeptide, a basigin (BSG) polypeptide, an immunoglobulin superfamily member 2 (IGSF2) polypeptide, an immunoglobulin superfamily member 3 (IGSF3) polypeptide, an immunoglobulin superfamily member 8 (IGSF8) polypeptide, an integrin beta-1 (ITGB1) polypeptide, an integrin beta-4 (ITGA4) polypeptide, a 4F2 cell-surface antigen heavy chain (SLC3A2) polypeptide, an ATP transporter polypeptide, a fragment thereof, or any combination thereof.
98 . The EV of claim 96 , wherein the scaffold lipid is selected from the group consisting of cholesterol and other sterols, fatty acids, phospholipids, lysolipids, vitamins, and combinations thereof.
99 . The EV of claim 85 , wherein the biologically active molecule comprises a vaccine antigen, a vaccine adjuvant, a small molecule, a proteolysis-targeting chimera (PROTAC), a stimulator of interferon genes protein (STING) agonist, a polynucleotide, an antisense oligonucleotide (ASO), a synthetic antineoplastic agent, a cytokine release inhibitor, a mammalian target of rapamycin (mTOR) inhibitor, an autotaxin inhibitor, a lysophosphatidic acid receptor 1 (LPA1) antagonist, or any combination thereof.
100 . The EV of claim 85 , further comprising a targeting moiety, a tropism moiety, an anti-phagocytic signal, or any combination thereof.
101 . The EV of claim 85 , wherein the EV is an exosome.
102 . A pharmaceutical composition comprising the EV of claim 85 and a pharmaceutically acceptable carrier.
103 . A method of treating a disease or disorder selected from the group consisting of a cancer, an inflammatory disorder, a neurodegenerative disorder, a central nervous diseases, and a metabolic disease in a subject in need thereof, comprising administering a therapeutically effective amount of the EV of claim 85 to the subject.
104 . A method of attaching a biologically active molecule to an EV comprising
(i) connecting a biologically active moiety to an anchoring moiety via a cleavable linker; and,
(ii) bringing the anchoring moiety in contact with the EV.