Mobilization of hematopoietic stem cells
Methods, processes, uses, and pharmaceutical compositions are provided herein for mobilizing hematopoietic progenitor cells and/or cancer stem cells from bone marrow into peripheral blood, comprising the administration of an effective amount of an inhibitor of GTPases, such as a Cdc-42 specific inhibitor alone or in combination with one or more additional agents. Specifically, methods are disclosed for mobilizing hematopoietic stem cells into a subject's peripheral blood. In particular, embodiments of the method involve specific inhibition of the Cdc42 GTPase to increase the numbers of hematopoietic stem cells into a subject's peripheral blood of a subject.
1. A method for facilitating hematopoietic reconstitution of peripheral blood precursor cells in a subject's hematopoietic organs, comprising: a) administering to the subject an effective amount of at least one Cdc42-specific inhibitor in the precursor cells; b) isolating mobilized peripheral blood precursor cells from the peripheral circulation of the subject; and c) infusing the isolated mobilized peripheral blood precursor cells into the subject, wherein said at least one Cdc42-specific inhibitor is selected from the group of compounds having the structure of Formula (I):
as a single isomer, a mixture of isomers, a racemic mixture of isomers, pharmaceutically acceptable salt, a solvate, metabolite or polymorph thereof, wherein:
Y is selected from the group consisting of OR 7 , NR 8 R 9 , and NNR 8 R 9 ;
R 7 is selected from the group consisting of C 1-6 alkyl, (CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, phenyl, C 1-6 alkyl substituted with up to 5 fluoro, and C 1-6 alkoxy substituted with up to 5 fluoro, said C 1-6 alkyl, (CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, phenyl are each optionally substituted with one or more substitutents each independently selected from the group consisting of halo, —CN, —OH, C 1-6 alkoxyl, heteroaryl, R 19 , and OR 20 ;
R 8 and R 9 are each separately a hydrogen, or separately selected from the group consisting of C 1-6 alkyl, C 3-7 cycloalkyl, and phenyl, said C 1-6 alkyl, C 3-7 cycloalkyl, and phenyl, each optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl, —(CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, R 19 , OR 20 , C 1-6 alkyl substituted with up to 5 fluoro, and C 1-6 alkoxy substituted with up to 5 fluoro, each optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl, and C 1-6 alkoxy; or R 8 and R 9 are optionally taken together with the nitrogen to which they are attached to form indolinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, each optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl, (CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, phenyl, C 1-6 alkyl substituted with up to 5 fluoro, and C 1-6 alkoxy substituted with up to 5 fluoro; or R 8 and R 2 come together to be C 1-3 alkyl linking together as a ring;
each u is independently 0, 1, 2, 3, or 4;
R 2 is a hydrogen, or selected from the group consisting of C 1-6 alkyl, C 3-7 cycloalkyl, and phenyl, said C 1-6 alkyl, C 3-7 cycloalkyl, and phenyl, each optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl, —(CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, phenyl, C 1-6 alkyl substituted with up to 5 fluoro, C 1-6 alkoxy substituted with up to 5 fluoro, and —O(CH 2 ) u phenyl optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl, and C 1-6 alkoxy; or R 8 and R 2 come together to be C 1-3 alkyl linking together as a ring;
R 3 , R 4 , R 5 and R 6 are each independently selected from the group consisting of: hydrogen, halo, cyano, nitro, hydroxy, C 1-6 alkyl, (CH 2 ) u C 3-7 cycloalkyl, —O(CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, phenyl, C 1-6 alkyl substituted with up to 5 fluoro, and C 1-6 alkoxy substituted with up to 5 fluoro, said C 1-6 alkyl, (CH 2 ) u C 3-7 cycloalkyl, —O(CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, and phenyl, each optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl, —(CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, phenyl, C 1-6 alkyl substituted with up to 5 fluoro, and C 1-6 alkoxy substituted with up to 5 fluoro, said phenyl optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl, —(CH 2 ) u C 3-7 cycloalkyl, C 2-6 alkenyl, C 1-6 alkoxy, hydroxy-C 1-6 alkyl, phenyl, C 1-6 alkyl substituted with up to 5 fluoro, and C 1-6 alkoxy substituted with up to 5 fluoro;
R 19 is aryl optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl optionally substituted with up to 5 fluoro, and C 1-6 alkoxy optionally substituted with up to 5 fluoro;
R 20 is hydrogen or aryl optionally substituted with one or more substituents each independently selected from the group consisting of halo, cyano, nitro, hydroxy, C 1-6 alkyl optionally substituted with up to 5 fluoro, and C 1-6 alkoxy optionally substituted with up to 5 fluoro; and
wherein when Y is NR 8 R 9 then R 8 and R 2 optionally come together to be C 1-3 alkyl linking together as a ring,
with the proviso when R 8 comes together with R 2 to be C 1-3 alkyl linking together as a ring then R 4 , is not substituted with hydroxyl.
2. The method of claim 1 , wherein the peripheral blood precursor cells are hematopoietic cells selected from the group consisting of progenitor cells and stem cells.
3. The method of claim 1 , further comprising administering a second agent prior to or concurrently with administering the Cdc42-specific inhibitor.
4. The method of claim 3 , wherein the second agent is selected from the group consisting of G-CSF, GM-CSF, IL-3, GM-CSF/IL-3 fusion proteins, FLK-2/FLT-3 ligand, stem cell factor, IL-6, IL-11, TPO, VEGF, AMD3100 and combinations thereof.
5. The method of claim 3 , wherein the second agent is G-CSF.
6. The method of claim 3 , wherein the second agent is AMD3100.
7. The method of claim 1 , wherein the hematopoietic cells are obtained from peripheral blood.
8. The method of claim 1 , wherein the hematopoietic cells are obtained from bone marrow.
9. The method of claim 1 , wherein the Cdc42-specific inhibitor is administered prior to, simultaneously with, or after a cancer therapy.
10. The method of claim 1 , wherein the at least one Cdc42-specific inhibitor negatively modulates an activity of Cdc42 protein selected from the group consisting of GTP binding, GDP binding, GEF binding, GTPase activity, integrin binding, coupling or binding of Cdc42 to a receptor, and coupling or binding of Cdc42 to a effector.
11. The method of claim 1 , wherein the at least one Cdc42-specific inhibitor is selected from the group consisting of