Methods and compositions related to dopaminergic neuronal cells

• US 8,815,592 B2
• Filed: 04/20/2011
• Issued: 08/26/2014
• Est. Priority Date: 04/21/2010
• Status: Active Grant

• Current Assignee: Research & Development Foundation
• Original Assignee: Research & Development Foundation
• Inventors: Krause, Karl-Heinz, Preynat-Seauve, Olivier
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US13/696,775
• Publication Number:
  US 20130149717A1
• Time in Patent Office: 1,224 Days
• Field of Search: 435/377, 435/325
• US Class Current: 435/377
• CPC Class Codes: A61K 31/4184 condensed with carbocyclic rings, e.g. benzimidazoles
• CPC Class Codes: A61K 31/423 condensed with carbocyclic rings
• CPC Class Codes: A61K 35/30 Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: C07D 235/26 Oxygen atoms
• CPC Class Codes: C07D 263/58 with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0619 Neurons

Abstract

A method of differentiation stem cells cells by contacting stem cells with a dopaminergic differentiation agent is provided in certain aspects. For example, the agent may comprise substituted benzoxazole. These methods and compositions may be used in toxicological screens, e.g., to evaluate the neurotoxicity of a test compound or treatment of neurological disorders.

Claims

1. A method of enhancing differentiation of stem cells into dopaminergic neural cells comprising inducing neural cell differentiation of stem cells in the presence of zoxazolamine (Zol) or a structural analog of Zol having the capacity to stimulate potassium channels for a sufficient amount of time to differentiate the stem cells into dopaminergic neural cells, wherein said stem cells are pluripotent stem cells or neural stem cells and wherein the presence of Zol or said analog increases the number of dopaminergic neurons compared to inducing neural cell differentiation of pluripotent cells or neural stem cells in the absence of Zol or said analog.

2. The method of claim 1, wherein the zoxazolamine or structural analog has a formula of:

3. The method of claim 1, wherein the zoxazolamine or structural analog is zoxazolamine.

4. The method of claim 1, wherein the zoxazolamine or structural analog is 5-chloro-2-hydroxybenzoxazole (chlorzoxazone).

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

6. The method of claim 1, further comprising evaluating the dopaminergic neurons for one or more characteristics of dopaminergic neuronal cells.

7. The method of claim 1, further comprising contacting the dopaminergic neurons with a test compound and determining if the test compound has a pharmacological or toxicological impact on said dopaminergic neuron.