Benzimidazole compounds differentiate a mammalian stem cell into mesodermal or cardiomyocyte cells

• US 9,012,217 B2
• Filed: 09/16/2009
• Issued: 04/21/2015
• Est. Priority Date: 09/17/2008
• Status: Active Grant

• Current Assignee: Human Biomolecular Research Institute, Sanford Burnham Prebys Medical Discovery Institute
• Original Assignee: Burnham Institute For Medical Research, Human Biomolecular Research Institute
• Inventors: Mercola, Mark, Dawson, Marcia, Cashman, John, Bushway, Paul J.
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US12/561,235
• Publication Number:
  US 20100159596A1
• Time in Patent Office: 2,043 Days
• US Class Current: 435/377
• CPC Class Codes: C07D 215/54 attached in position 3
• CPC Class Codes: C07D 235/06 with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 2
• CPC Class Codes: C07D 279/20 with hydrogen atoms directly attached to the ring nitrogen atom
• CPC Class Codes: C07D 279/22 with carbon atoms directly attached to the ring nitrogen atom
• CPC Class Codes: C07D 279/28 with other substituents attached to the ring system
• CPC Class Codes: C07D 401/04 directly linked by a ring-member-to-ring-member bond
• CPC Class Codes: C07D 401/10 linked by a carbon chain containing aromatic rings
• CPC Class Codes: C07D 401/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07D 401/14 containing three or more hetero rings
• CPC Class Codes: C07D 403/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07D 405/04 directly linked by a ring-member-to-ring-member bond
• CPC Class Codes: C07D 405/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07D 409/04 directly linked by a ring-member-to-ring-member bond
• CPC Class Codes: C07D 491/04 Ortho-condensed systems
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

Methods and small molecule compounds for stem cell differentiation are provided. One example of a class of compounds that may be used is represented by the compound having the structure IA or IB in the form of free base or a pharmaceutically acceptable salt, hydrate, solvate or N-oxide thereof:

R₁ is independently hydrogen or (C₁-C₆)alkyl; R₂ is independently hydrogen, (C₁-C₆)alkyl, aryl, or heteroaryl; R_2′ is independently hydrogen, (C₁-C₆)alkyl, CF₃ or C₂F₅; R₃ is independently (C₁-C₆)alkyl, aryl, 2-tetrahydrofurylmethyl, an aliphatic tertiary amine, or 4-methoxybenzyl; or R₂ and R₃ may be joined together to form a 5 or 6 member ring lactone; R₄ is independently hydrogen, (C₁-C₆)alkyl, a 2- or 4-R₅-substituted aromatic ring selected from a 4-R₅-phenyl or a 2-R₅-5-pyridyl, aryl, heteroaryl, aliphatic tertiary amine or halogen; and R₅, R_5′, R₆, R_6′, R₇, R_7′, are each independently hydrogen, (C₁-C₆)alkyl, aryl, optionally substituted phenyl, heteroaryl, a heterocyclic ring, an aliphatic tertiary amine, or halogen.

Claims

1. A method for producing mammalian mesodermal or cardiomyocyte cells from mammalian stem cells, comprising contacting within the same suitable medium Activin A and the stem cells with a compound of structure II in the form of a free base or a pharmaceutically acceptable salt, hydrate, solvate or N-oxide thereof:

2. The method of claim 1, wherein the compound of structure II has formula:

3. The method of claim 1, wherein the compound of structure II has structure IIA, IIB, IIC, IID, IIE, IIF, IIG, or IIM in the form of a free base or a pharmaceutically acceptable salt, hydrate, solvate or N-oxide thereof:

4. The method of claim 1, wherein the compound of structure II has the formula:

5. The method of claim 1, wherein the contacting is for a period of time between about 24 hours to 192 hours.

6. The method of claim 5, wherein the contacting is between about 48 hours to 144 hours.

7. The method of claim 1, wherein cells produced from mammalian stem cells are cardiomyocytes.

8. The method of claim 1, wherein the cells produced from mammalian stem cells are mesodermal cells.

9. The method of claim 1, further comprising contacting the cells with a Wnt protein.

10. The method of claim 9, wherein the Wnt protein is Wnt3a.

11. The method of claim 1, wherein the stem cells are embryonic stem cells, induced pluripotent stem cells or adult stem cells.