EXTRACELLULAR VESICLE COMPOSITIONS
The present disclosure relates to compositions for the storage and administration of extracellular vesicles (e.g., exosomes) that can comprise a scaffold protein and one or more (e.g., 1, 2, 3, 4, 5 or more) exogenous biologically active moieties. Also provided herein are methods for producing the exosomes and methods for using the exosomes to treat and/or prevent a range of medical disorders.
1 . A pharmaceutical composition comprising
(a) an extracellular vesicle;
(b) a saccharide;
(c) sodium chloride;
(d) a potassium phosphate; and
(e) a sodium phosphate,
wherein the composition is in a solution at a pH of 7.1 to 7.3.
2 . The composition of claim 1 , wherein the extracellular vesicle is an exosome.
3 . The composition of claim 1 , wherein the composition is capable of being stored for at least about 4 hours at a temperature of 4° C.
4 - 5 . (canceled)
6 . The composition of claim 1 , wherein the composition has a pH of 7.2.
7 . (canceled)
8 . The composition of claim 1 , wherein the composition has (i) reduced aggregates, (ii) improved stability of the EV, (iii) improved integrity of the EV architecture, and (iv) improved stability of engineered proteins contained on or in EVs.
9 - 10 . (canceled)
11 . The composition of claim 1 , wherein the saccharide comprises lactose, glucose, sucrose, trehalose, and/or combinations thereof.
12 - 17 . (canceled)
18 . The composition of claim 1 , wherein the composition has a conductivity between about 6 mS/cm+/−10% and about 10 mS/cm+/−10%.
19 - 21 . (canceled)
22 . The composition of claim 1 , wherein the sodium chloride is present in the composition at a concentration of between about 10 mM and about 134 mM.
23 - 32 . (canceled)
33 . The composition of claim 1 , wherein the potassium phosphate is present in the composition at a concentration of between about 1 mM to about 20 mM, between about 2 mM to about 19 mM, between about 3 mM to about 18 mM, between about 4 mM to about 17 mM, between about 5 mM to about 16 mM, or between about 5 mM to about 15 mM.
34 - 37 . (canceled)
38 . The composition of claim 1 , wherein the potassium phosphate is potassium phosphate monobasic.
39 . The composition of claim 1 , wherein the sodium phosphate is present in the composition at a concentration of between about 10 mM to about 30, between about 11 mM to about 29 mM, between about 12 mM to about 28 mM, between about 13 mM to about 27 mM, or between about 14 mM to about 26 mM.
40 - 44 . (canceled)
45 . The composition of any one of claims 1 to 44 , further comprising an anti-oxidant, wherein the anti-oxidant comprises D-methionine, L-methionine, ascorbic acid, erythorbic acid, Na ascorbate, thioglycerol, cysteine, acetylcysteine, cystine, dithioerythreitol, glutathione, tocopherols, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), sodium bisulphate, sodium dithionite, A-Tocopherol, γ-Tocopherol, propyl gallate, ascorbyl palmitate, sodium metabisulfite, thiourea, sodium thiosulfate, propyl gallate, and sodium thioglycolate.
46 . (canceled)
47 . The composition of claim 1 , wherein the composition:
(i) is not lyophilized,
(ii) does not comprise a chelating agent,
(iii) does not comprise albumin, or
(iv) any combination of (i) to (iii).
48 - 49 . (canceled)
50 . The composition of claim 1 , comprising
(1)
a. a sucrose at a concentration of about 2.5% to about 5% w/v,
b. sodium chloride at a concentration of about 50 mM to 150 mM,
c. a potassium phosphate monobasic at a concentration of about 5 mM, and
d. a sodium phosphate dibasic heptahydrate at a concentration of about 15 mM;
wherein the composition is in a solution at a pH of 7.2 and at a conductivity of 7.23 mS/cm+/−10%; or
(2)
a. a sucrose at a concentration of about 5% w/v,
b. sodium chloride at a concentration of about 40 mM,
c. a potassium phosphate monobasic at a concentration of about 15 mM, and
d. a sodium phosphate dibasic heptahydrate at a concentration of about 27 mM,
wherein the composition is in a solution at a pH of 7.2 and at a conductivity of 8.8 mS/cm+/−10%.
51 - 75 . (canceled)
76 . A method of preparing the pharmaceutical composition of claim 1 comprising combining:
(a) an extracellular vesicle;
(b) a saccharide;
(c) sodium chloride;
(d) a potassium phosphate; and
(e) a sodium phosphate.
77 - 82 . (canceled)
83 . A method of treating a disease or a condition in a subject in need thereof comprising administering to the subject the composition of claim 1 .
84 - 89 . (canceled)
90 . The composition of claim 1 , wherein the extracellular vesicle comprises:
(i) an antisense oligonucleotide (ASO),
(ii) a STING agonist, or
(iii) an IL-12 moiety.
91 . The composition of claim 1 , comprising:
(a) Extracellular vesicles comprising an ASO, wherein the ASO comprises a nucleic acid sequence selected from SEQ ID NOs: 91-193;
(b) Sucrose at a concentration of 2.5% to about 5%;
(c) Sodium chloride at a concentration of 50 mM to 150 mM;
(d) Potassium phosphate monobasic at a concentration of 5 mM; and
(e) Sodium phosphate dibasic at a concentration of 15 mM.
92 - 93 . (canceled)
94 . The composition of claim 1 , comprising:
(a) Extracellular vesicles;
(b) Sucrose at a concentration of about 5%;
(c) Sodium chloride at a concentration of about 50 mM;
(d) Potassium phosphate monobasic at a concentration of about 5 mM;
(e) Sodium phosphate dibasic at a concentration of about 15 mM, and
(f) wherein the pH of the composition is about 7.2;
wherein the extracellular vesicles comprise an IL-12 moiety.
95 - 115 . (canceled)
116 . The composition of claim 91 , wherein
(i) the ASO comprises the nucleic acid sequence set forth in SEQ ID NO: 144, 145, 193, or 185;
(ii) the ASO is associated with the extracellular vesicles by a linker, comprising cholesterol, tocopherol, a fatty acid, or any combination thereof; or
(iii) any combination of (i) and (ii).