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US 9,005,968 B2Granted
Induction of pluripotent cells
AssigneeThe Scripps Research Institute
InventorsTongxiang Lin, Sheng Ding
Life of this patent
Estimating the term…
Abstract
The slow kinetics and low efficiency of reprogramming methods to generate human induced pluripotent stem cells (iPSCs) impose major limitations on their utility in biomedical applications. Here we describe a chemical approach that dramatically improves (>200 fold) the efficiency of iPSC generation from human fibroblasts, within seven days of treatment. This will provide a basis for developing safer, more efficient, non-viral methods for reprogramming human somatic cells.
Claims24
A composition comprising:
- an activin receptor-like kinase 5 (ALK5) inhibitor;
- a MAP/ERK kinase (MEK) inhibitor;
- and
- a Rho kinase (ROCK) inhibitor, wherein the ROCK inhibitor has the structure:
- wherein L2 is substituted or unsubstituted C1-C10 alkylene;
- y is an integer from 0 to 3;
- z is an integer from 0 to 5;
- X is —N═, —CH═, or —CR5=;
- R1 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- R3, R4, and R5 are independently —CN, —S(O)nR6, —NR7R8, —C(O)R9, —NR10—C(O)R11, —NR12—C(O)—OR13, —C(O)NR14R15, —NR16S(O)2R17, —OR18, —S(O)2NR19, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, wherein n is an integer from 0 to 2, wherein if z is greater than 1, two R3 moieties are optionally joined together to form a substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- and
- R6, R7, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, R18 and R19 are independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- or a racemate, diastereomer, tautomer, or a geometric isomer thereof, or a pharmaceutically acceptable salt thereof.
10 more elements
A mixture comprising:
- one or more mammalian cells;
- an ALK5 inhibitor;
- a MEK inhibitor;
- and
- a ROCK inhibitor, wherein the ROCK inhibitor has the structure:
- wherein L2 is substituted or unsubstituted C1-C10 alkylene;
- y is an integer from 0 to 3;
- z is an integer from 0 to 5;
- X is —N—, —CH—, or —CR5=;
- R1 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- R3, R4, and R5 are independently —CN, —S(O)nR6, —NR7R8, —C(O)R9, —NR10—C(O)R11, —NR12—C(O)—OR13, —C(O)NR14R15, —NR16S(O)2R17, —OR18, —S(O)2NR19, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, wherein n is an integer from 0 to 2, wherein if z is greater than 1, two R3 moieties are optionally joined together to form a substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- and
- R6, R7, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, R18 and R19 are independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- or a racemate, diastereomer, tautomer, or a geometric isomer thereof, or a pharmaceutically acceptable salt thereof.
11 more elements
A method of inducing non-pluripotent mammalian cells into induced pluripotent mammalian stem cells, comprising:
- (a) introducing Oct-3/4, Klf4, Sox2, and optionally c-Myc into the non-pluripotent cells;
- and
- (b) contacting the non-pluripotent cells with an ALK5 inhibitor, a MEK inhibitor, and a ROCK inhibitor, wherein the ROCK inhibitor has the structure:
- wherein L2 is substituted or unsubstituted C1-C10 alkylene;
- y is an integer from 0 to 3;
- z is an integer from 0 to 5;
- X is —N═, —CH═, or —CR5=;
- R1 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- R3, R4, and R5 are independently —CN, —S(O)nR6, —NR7R8, —C(O)R9, —NR10—C(O)R11, —NR12—C(O)—OR13, —C(O)NR14R15, —NR16S(O)2R17, —OR18, —S(O)2NR19, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, wherein n is an integer from 0 to 2, wherein if z is greater than 1, two R3 moieties are optionally joined together to form a substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- and
- R6, R7, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, R18 and R19 are independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
- or a racemate, diastereomer, tautomer, or a geometric isomer thereof, or a pharmaceutically acceptable salt thereof.
9 more elements
Dependencies are inferred from the claim text.