IMPROVED FORMULATIONS FOR IMMUNE CELLS
We disclose various improvements for compositions of T cells, including the use of lower levels of serum albumin and of cryoprotectants such as dimethyl sulfoxide.
1 . A composition comprising T cells and serum albumin and dimethyl sulfoxide (DMSO), wherein the composition comprises <2.5% (v/v) serum albumin, and wherein DMSO is present at a ≥5-fold excess (v/v) compared to the serum albumin.
2 . The composition of claim 1 , comprising <2.2% (v/v), <2% (v/v), <1% (v/v), or about 0.5% (v/v) serum albumin.
3 . The composition of claim 1 or claim 2 , wherein DMSO is present at a 5-fold to 30-fold excess (v/v) compared to the serum albumin.
4 . The composition of claim 3 , wherein the excess is from 5-fold to 15-fold, from 6-fold to 12-fold, from 8-fold to 11-fold, or about 10-fold.
5 . A composition comprising T cells, serum albumin, and DMSO, wherein the concentration of serum albumin (v/v) is from 0.1-1% and the concentration of DMSO (v/v) is 3-fold to 30-fold higher than the concentration of serum albumin.
6 . The composition of claim 5 , wherein the serum albumin concentration is <1% (v/v), and the DMSO concentration is from 8-fold to 11-fold higher than the serum albumin concentration.
7 . The composition of claim 5 or claim 6 , wherein concentration of DMSO is <10% (v/v).
8 . The composition of claim 7 , wherein the concentration of DMSO is from 2-8% (v/v), from 4-7% (v/v), from 5-6% (v/v), or about 5% (v/v).
9 . A composition comprising T cells, serum albumin, and DMSO, wherein the concentration of serum albumin (v/v) is 0.1-1% and the concentration of DMSO is 5-7% (v/v).
10 . The composition of claim 9 , wherein the concentration of serum albumin (v/v) is between 0.2-0.8%, between 0.3-0.7%, between 0.4-0.6%, or about 0.5%.
11 . The composition of any one of claims 1 to 10 , having between 1×10 6 and 40×10 6 T cells/mL.
12 . The composition of any one of claims 1 to 11 , in cryopreserved form.
13 . A composition comprising T cells and serum albumin, wherein the composition comprises about 0.15% (v/v) serum albumin.
14 . The composition of claim 13 , also including dimethyl sulfoxide (DMSO).
15 . The composition of claim 14 , wherein DMSO is present at a 3-fold to 30-fold excess (v/v) compared to the serum albumin.
16 . The composition of claim 15 , wherein the excess is from 3-fold to 15-fold, from 6-fold to 12-fold, from 8-fold to 11-fold, or about 10-fold.
17 . A composition comprising T cells, serum albumin, and DMSO, wherein the concentration of serum albumin (v/v) is from 0.03-0.6% and the concentration of DMSO (v/v) is 3-fold to 30-fold higher than the concentration of serum albumin.
18 . The composition of claim 17 , wherein the serum albumin concentration is <0.3% (v/v), and the DMSO concentration is from 8-fold to 11-fold higher than the serum albumin concentration.
19 . The composition of claim 17 or claim 18 , wherein concentration of DMSO is <3% (v/v).
20 . The composition of claim 7 , wherein the concentration of DMSO is from 0.6-2.4% (v/v), from 0.8-2.2% (v/v), from 1.5-2% (v/v), or about 1.8% (v/v).
21 . A composition comprising T cells and DMSO, wherein the composition comprises <0.15% (v/v) DMSO.
22 . A composition comprising T cells, serum albumin, and DMSO, wherein the concentration of serum albumin is 0.03-0.3% (v/v) and the DMSO concentration is 0.15-0.20% (v/v).
23 . The composition of any one of claims 13 to 22 , having between 0.3×10 6 and 12×10 6 T cells/mL.
24 . The composition of any preceding claim, wherein the T cells are genetically modified T cells, such as CAR-T cells.
25 . The composition of any preceding claim, wherein the T cells are human T cells and the serum albumin is human serum albumin.
26 . A process for preparing the composition of any one of claims 13 to 25 , comprising the step of diluting a composition of any one of claims 1 to 12 .
27 . The process of claim 26 , wherein dilution is performed with an electrolyte solution.
28 . The process of claim 27 , wherein the electrolyte solution is normal saline.
29 . The process of claim 27 , wherein the electrolyte solution is a sterile, nonpyrogenic, isotonic solution with a pH of 6.5-8.0 that has about 5.26 g/L NaCl, about 5.02 g/L sodium gluconate, about 3.68 g/L sodium acetate trihydrate, about 0.37 g/L KCl, and about 0.3 g/L MgCl 2 .6H 2 O.
30 . A method of treating a subject, comprising a step of administering a composition of any one of claims 1 to 25 to a patient in need thereof.
31 . A container comprising a suspension of T cells, serum albumin and dimethyl sulfoxide (DMSO), wherein the composition comprises <2.5% (v/v) serum albumin, and wherein DMSO is present at a ≥5-fold excess (v/v) compared to the serum albumin, and wherein the suspension is to be administered to a patient.
32 . The container of claim 31 , wherein the suspension comprises <2.2% (v/v), <2% (v/v), <1% (v/v), or about 0.5% (v/v) serum albumin.
33 . The container of claim 31 or claim 32 , wherein DMSO is present at a 5-fold to 30-fold excess (v/v) compared to the serum albumin.
34 . The container of claim 33 , wherein the excess is from 5-fold to 15-fold, from 6-fold to 12-fold, from 8-fold to 11-fold, or about 10-fold.
35 . A container comprising a suspension of T cells, serum albumin, and DMSO, wherein the concentration of serum albumin (v/v) is from 0.1-1% and the concentration of DMSO (v/v) is 3-fold to 30-fold higher than the concentration of serum albumin, and wherein the suspension is to be administered to a patient.
36 . The container of claim 35 , wherein the serum albumin concentration is <1% (v/v), and the DMSO concentration is from 8-fold to 11-fold higher than the serum albumin concentration.
37 . The container of claim 35 or claim 36 , wherein concentration of DMSO is <10% (v/v).
38 . The container of claim 37 , wherein the concentration of DMSO is from 2-8% (v/v), from 4-7% (v/v), from 5-6% (v/v), or about 5% (v/v).
39 . A container comprising a suspension of T cells, serum albumin, and DMSO, wherein the concentration of serum albumin (v/v) is 0.1-1% and the concentration of DMSO is 5-7% (v/v), and wherein the suspension is to be administered to a patient.
40 . The container of claim 39 , wherein the concentration of serum albumin (v/v) is between 0.2-0.8%, between 0.3-0.7%, between 0.4-0.6%, or about 0.5%.
41 . The container of any one of claims 31 to 40 , having between 1×10 6 and 40×10 6 T cells/mL.
42 . The composition of any one of claims 31 to 41 , in cryopreserved form.
43 . A method of treating a subject, comprising a step of administering to a patient in need thereof the composition of any one of claims 13 to 25 by intravenous infusion.
44 . The method of claim 43 , wherein the intravenous infusion time is between 15 and 120 minutes.
45 . The method of claim 43 , wherein the intravenous infusion time is up to 30 minutes.
46 . The method of any one of claims 41 - 43 , wherein the infusion volume is between 50 and 100 mL.
47 . The method of any one of claims 1 - 4 , wherein the infusion volume is about 50 mL.
48 . The composition, method, process, or container of any preceding claim, wherein the T cells comprise a polynucleotide that encodes a chimeric signaling polypeptide, wherein the chimeric signaling polypeptide comprises:
(i) a costimulatory polypeptide cytoplasmic signaling region;
(ii) a truncated MyD88 polypeptide region lacking the TIR domain;
(iii) a truncated MyD88 polypeptide region lacking the TIR domain and a costimulatory polypeptide cytoplasmic signaling region; or
(iv) a truncated MyD88 polypeptide region lacking the TIR domain and a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain.
49 . The composition, method, process, or container of claim 48 , wherein costimulatory polypeptide cytoplasmic signaling region is a signaling region that activates the signaling pathways activated by MyD88, CD40 and/or MyD88-CD40 fusion chimeric polypeptide.
50 . The composition, method, process, or container of any of the preceding claim, wherein the T cells comprises a polynucleotide encoding an inducible chimeric pro-apoptotic polypeptide.
51 . A kit comprising:
(a) a container comprising an interior space; and
(b) a composition contained within the interior space, wherein the composition comprises:
(i) T cells in an amount between about 1×10 6 and 40×10 6 (e.g., about 5×10 6 ) cells/ml;
(ii) serum albumin in a concentration of about 0.1%-0.6% v:v;
(iii) DMSO in a concentration of about 5%-7% v:v; and
(iv) a pharmaceutically acceptable carrier for human injection.
52 . The kit of claim 51 , wherein the container comprises a bag, e.g., a cryopreservation bag.
53 . The kit of claim 52 , wherein the container further comprises a bag spike in fluidic communication with the interior space.
54 . The kit of claim 51 , wherein the T cells are genetically engineered.
55 . The kit of claim 51 , wherein the serum albumin is human serum albumin.
56 . The kit of claim 51 , wherein the serum albumin is present in concentration of about 0.5% v:v.
57 . The kit of claim 51 , wherein DMSO is present in a concentration of about 5% v:v.
58 . The kit of claim 51 , wherein the pharmaceutically acceptable carrier comprises sodium chloride, sodium gluconate sodium, acetate trihydrate, potassium chloride, and magnesium chloride
59 . The kit of claim 51 , wherein the pharmaceutically acceptable carrier comprises Pharma Lyte or Plasma Lyte.
60 . The kit of claim 51 , wherein the composition has a volume of about 15 ml.
61 . A kit comprising:
(a) a container comprising an interior space; and
(b) a composition contained within the interior space, wherein the composition comprises:
(i) T cells in an amount between about 15×10 6 cells and about 600×10 6 (e.g., about 75×10 6 ) cells;
(ii) serum albumin in an amount between about 0.03% and about 1.8% serum albumin, e.g., about 1.5% v:v;
(iii) DMSO in a concentration of about 1.5%-2.1 v:v (e.g., about 1.5% v:v); and
(iv) a pharmaceutically acceptable carrier for human injection.
62 . The kit of claim 61 , wherein the container comprises a bag, optionally including a tube comprising a bag spike, wherein the tube is in fluidic communication with the interior space.
63 . The kit of claim 61 , wherein the composition has a volume of about 50 ml.
64 . A composition as described herein for use in treating a disease, e.g., cancer.
65 . A use of a composition as described herein for treating a disease, e.g., cancer.
66 . A use of a composition as described herein in the preparation of a medicament for treating a disease, e.g., cancer.