CBL inhibitors and compositions for expansion of immune cells
Methods and compositions using novel Cbl inhibitors enhancing expansion of immune cells to increase the efficacy of cell-based immunotherapeutics are disclosed. Also provided are cell-based immunotherapy methods and compositions.
1 . A method of producing expanded immune cells, comprising the step of:
expanding immune cells in vitro in the presence of a Cbl inhibitor in culture under conditions wherein at least 10 9 expanded immune cells are produced in a first expansion,
wherein the immune cells are tumor infiltrating lymphocytes,
wherein no expansion steps are performed before the first expansion, and
wherein the Cbl inhibitor is a compound of Formula (A):
or a tautomer thereof, or a pharmaceutically acceptable salt of any of the foregoing, wherein:
R 1 is H, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, 3-to 6-membered heterocyclyl, C 1 -C 6 alkyl-(C 3 -C 6 cycloalkyl), C 1 -C 6 alkyl-(3-to 6-membered heterocyclyl);
Z 1 is CH or N;
Z 2 is CH or N;
Z 3 is CH or N;
X is CH or N;
R 2 is H, halo, C 3 -C 6 cycloalkyl, —NH-(3-to 6-membered heterocyclyl), —NH—(C 1 -C 6 alkyl),
—NH—(C 3 -C 6 cycloalkyl), —O-(3-to 6-membered heterocyclyl), —O—(C 1 -C 6 alkyl), or
—O—(C 3 -to C 6 cycloalkyl);
R 3a is H, halo, or C 1 -C 6 alkyl;
R 3b is H, halo, C 1 -C 6 alkyl, or C 3 -C 6 cycloalkyl; or R 3a and R 3b are taken together with the carbon atom to which they are attached to form a 4- to 8-membered heterocyclyl or a C 3 -C 6 cycloalkyl, wherein the heterocyclyl or cycloalkyl are optionally substituted by 1-3 R 12 groups;
n is 0 or 1;
Y is C(R 11a )(R 11b ) or S, with the proviso that if either or both of R 3a and R 3b are halo, Y is C(R 11a )(R 11b );
Q is CH or N;
R 4 is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, or C 3 -C 6 cycloalkyl;
R 10 is H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, or C 3 -C 6 cycloalkyl;
R 11a and R 11b are independently H, halo, C 1 -C 6 alkyl, or C 1 -C 6 haloalkyl; or R 11a and R 11b are taken together with the carbon atom to which they are attached to form a C 3 -C 6 cycloalkyl; or R 3b and R 11a are taken together with the carbon atoms to which they are attached to form a C 3 -C 6 cycloalkyl optionally substituted by C 1 -C 6 alkyl; and
each R 12 is independently C 1 -C 6 alkyl, halo, hydroxy, —O(C 1 -C 6 alkyl), —CN, C 1 -C 6 alkyl-CN, C 1 -C 6 alkyl-OH, or C 1 -C 6 haloalkyl; wherein two geminal R 12 groups can be taken together with the carbon atom to which they are attached to form a spiro C 3 -C 4 cycloalkyl.
2 . The method of claim 1 that comprises the first expansion step and no further expansion steps.
3 . The method of claim 1 that comprises the first expansion step and one or more further expansions.
4 . The method of claim 3 wherein the first expansion is for 8-12 days.
5 . The method of claim 3 wherein the second expansion is for 8-12 days and there are no further expansions.
6 . The method of claim 1 , wherein the Cbl inhibitor is added to the culture every 3-5 days.
7 . The method of claim 1 , wherein the Cbl inhibitor is added to the culture on or about a day 0, a day 4, and a day 7 of the first expansion relative to culture initiation of the first expansion.
8 . The method of claim 1 , further comprising a second expansion step, wherein the Cbl inhibitor is added to the culture on or about a day 0 and a day 5 of the second expansion step relative to culture initiation of the second expansion step.
9 . The method of claim 1 , wherein the culture further comprises IL-2.
10 . The method of claim 1 , further comprising a second expansion, wherein IL-2 is added to the first expansion at a day 0 and to the second expansion at a day 0 and a day 5 relative to the first expansion and the second expansion, respectively.
11 . The method of claim 1 , wherein the culture does not comprise a component that binds to a surface receptor of the immune cells.
12 . The method of claim 1 , wherein the culture does not comprise anti-CD3 antibody or OKT3.
13 . The method of claim 1 , wherein the culture does not comprise added feeder cells.
14 . The method of claim 1 , wherein the culture does not comprise added irradiated peripheral blood mononuclear cells.
15 . The method of claim 1 , wherein the culture does not comprise added 4-IBB agonist.
16 . The method of claim 1 , wherein the immune cells are harvested from a donor.
17 . The method of claim 1 , wherein the immune cells are from a tumor or a tumor fragment.
18 . The method of claim 1 that produces at least 10 10 or 10 11 expanded immune cells.
19 . The method of claim 1 , wherein the expanded immune cells are capable of use in therapy without further expansion.
20 . The method of claim 1 , wherein the expanded immune cells secrete a proinflammatory cytokine.
21 . The method of claim 20 , wherein said proinflammatory cytokine is IL-2 or IFNγ.
22 . The method of claim 1 , further comprising a second expansion, wherein the length of the first expansion is 0-4 days, 0-7 days, 0-11 days, 0-14 days, or 0-30 days, and/or wherein the length of the second expansion is 0-4 days, 0-7 days, 0-11 days, 0-14 days, or 0-30 days.
23 . The method of claim 1 , wherein the immune cells are expanded for 0-4 days, 0-7 days, 0-11 days, 0-14 days, or 0-30 days.
24 . The method of claim 1 , wherein the immune cells are expanded in vitro in a culture vessel comprising a gas permeable rapid expansion cell culture membrane.
25 . The method of claim 1 , wherein the Cbl inhibitor is at a concentration of 0.5-50 μM.
26 . The method of claim 1 , wherein the Cbl inhibitor inhibits activity of an enzyme selected from the group consisting of cCbl, Cbl-b, and Cbl-c.
27 . The method of claim 1 , wherein the Cbl inhibitor is a small molecule selected from the group consisting of compounds 101-129:
Compound
No.
Structure
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
28 . A composition comprising expanded immune cells produced by the method of claim 1 .
29 . A method of treating T cell dysfunction in a patient in need thereof, comprising administering to the patient an effective amount of the composition of claim 28 .
30 . A method of treating cancer in a patient in need thereof, comprising administering to the patient an effective amount of the composition of claim 28 .
31 . The method of claim 30 , further comprising administering to the patient a Cbl inhibitor, or a salt or pharmaceutically acceptable composition thereof, in an amount effective to enhance the treatment.