EXTRACELLULAR VESICLES COMPRISING STING-AGONIST
Provided herein are compositions comprising EV, e.g., exosome, encapsulated STING agonists and methods of producing the compositions described. Also provided herein are methods of modulating an immune response via administration of a therapeutic amount of EV, e.g., exosomes encapsulating STING agonists. The immune response may be an IENβ response or activation of myeloid dendritic cells (mDCs). Also provided herein are methods of modulating an immune response that does not induce systemic inflammation via administration of exosomes encapsulating STING agonists.
1 . A composition comprising an extracellular vesicle and a stimulator of interferon genes protein (STING) agonist.
2 . The composition of claim 1 , wherein the extracellular vesicle is an exosome, a nanovesicle, an apoptotic body, a microvesicle, a lysosome, an endosome, a liposome, a lipid nanoparticle, a micelle, a multilamellar structure, a revesiculated vesicle, or an extruded cell.
3 . (canceled)
4 . The composition of claim 1 , wherein the STING agonist is associated with the extracellular vesicle.
5 . The composition of claim 4 , wherein the STING agonist is encapsulated within the extracellular vesicle.
6 . (canceled)
7 . The composition of claim 5 , wherein the extracellular vesicle overexpresses a prostaglandin F2 receptor negative regulator (PTGFRN) protein or a fragment thereof.
8 - 13 . (canceled)
14 . The composition of claim 7 , wherein the extracellular vesicle further comprises a ligand, a cytokine, or an antibody.
15 - 17 . (canceled)
18 . The composition of claim 14 , wherein the STING agonist is a cyclic dinucleotide, a non-cyclic dinucleotide, or a lipid-binding tag.
19 - 25 . (canceled)
26 . The composition of claim 18 , wherein the STING agonist comprises:
wherein
X 1 is H, OH, or F;
X 2 is H, OH, or F;
Z is OH, OR 1 , SF 1 or SRI, wherein:
i) R 1 is Na or NH 4 , or
ii) R 1 is an enzyme-labile group which provides OH or SH in vivo such as pivaloyloxymethyl;
B1 and B2 are bases chosen from:
With the proviso that:
in Formula (I): X 1 and X 2 are not OH,
in Formula (II): when X 1 and X 2 are OH, B 1 is not Adenine and B 2 is not Guanine, and
in Formula (III): when X 1 and X 2 are OH, B 1 s not Adenine, B 2 is not Guanine and Z is not OH, or a pharmaceutically acceptable salt thereof.
27 . The composition of claim 26 , wherein the STING agonist is selected from the group consisting of:
and a pharmaceutically acceptable salt thereof.
28 . (canceled)
29 . The composition claim 1 , wherein the extracellular vesicle associated with the STING agonist exhibits one or more of the following characteristics:
(i) activates dendritic cells, e.g., myeloid dendritic cells;
(ii) activates monocyte cells at a lesser degree than the STING agonist alone (“free STING agonist”);
(iii) does not activate monocyte cells;
(iv) has a wider therapeutic index compared to the free STING agonist;
(v) has less systemic toxicity than the free STING agonist;
(vi) has less immune cell killing than the free STING agonist;
(vii) has higher cell selectivity than the free STING agonist;
(viii) provides tumor protective immunity at a dose lower than the free STING agonist;
(ix) induce a specific cellular response in vivo in antigen-presenting cells, e.g., dendritic cells,
(x) is capable of inducing an immune response at a distal region after a local administration; and
(xi) is capable of being dosed at a lower level than the free STING agonist.
30 . The composition of claim 1 , wherein the extracellular vesicle associated with the STING agonist, when administered to a mammal, (a) does not deplete T cells and/or macrophages in the mammal or (b) depletes T cells and/or macrophages in the mammal at a lesser degree than the free STING agonist.
31 . (canceled)
32 . A pharmaceutical composition comprising the composition of claim 1 and a pharmaceutically acceptable carrier.
33 . A kit comprising the composition of claim 1 and instructions for use.
34 . A method of producing an EV, e.g., exosome, comprising a STING agonist, the method comprising:
a. Obtaining an EV, e.g., exosome;
b. Mixing the EV, e.g., exosome with a STING agonist in a solution;
c. Incubating the mixture of the EV, e.g., exosome and the STING agonist in a solution comprising a buffer under suitable conditions; and
d. Purifying the EV, e.g., exosome comprising the STING agonist.
35 - 44 . (canceled)
45 . A method of inducing or modulating an immune response and/or an inflammatory response in a subject in need thereof, the method comprising administering to the subject a pharmaceutically effective amount of the pharmaceutical composition of claim 32 .
46 . A method of treating a tumor in a subject in need thereof, the method comprising administering to the subject the pharmaceutical composition of claim 32 .
47 - 54 . (canceled)
55 . The method of claim 45 , wherein the administration is parenterally, orally, intravenously, intramuscularly, intra-tumorally, intraperitoneally, or via any other appropriate administration route.
56 . (canceled)
57 . The method of claim 45 , wherein the immune response is an anti-tumor response.
58 - 59 . (canceled)
60 . The method of claim 46 , further comprising administering an additional therapeutic agent.
61 - 64 . (canceled)
65 . The composition of claim 26 , wherein the STING agonist is
or a pharmaceutically acceptable salt thereof.