Methods of treating ischemic spasticity

• US 9,744,194 B2
• Filed: 11/30/2015
• Issued: 08/29/2017
• Est. Priority Date: 11/17/2004
• Status: Active Grant

• Current Assignee: Neuralstem Incorporated (City of Tianjin (China)), Regents of the University of California (University of California)
• Original Assignee: Neuralstem Incorporated (City of Tianjin (China)), Regents of the University of California (University of California)
• Inventors: Johe, Karl K., Hazel, Thomas G., Marsala, Martin, Kakinohana, Osamu
• Primary Examiner(s): Ketter, Jim
• Application Number:
  US14/954,105
• Publication Number:
  US 20160228471A1
• Time in Patent Office: 638 Days
• Field of Search: None
• 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: A61K 9/0085 Brain, e.g. brain implants; Spinal cord
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/113 Acidic fibroblast growth factor (aFGF, FGF-1)
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/148 Transforming growth factor alpha [TGF-a]
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

The invention relates generally to methods of treating spasticity, rigidity, or muscular hyperactivity conditions by introducing a portion of an expanded population of neural stem cells into an area of a recipient spinal cord.

Claims

1. A method of treating neurodegenerative conditions in a subject in need thereof comprising:

a) obtaining neural stem cells of human origin;

b) expanding in vitro the neural stem cells in a dispersed adherent culture to form an expanded population of neural stem cells, wherein expanding the neural stem cells comprises;

i.) providing at least one extracellular protein to a culture vessel;

ii.) culturing the dissociated neural stem cells in said culture vessel in the absence of serum;

iii.) adding to the culture vessel at least one growth factor; and

iv.) passaging the cultured cells prior to confluence;

c) concentrating the expanded population of neural stem cells to form concentrated neural stem cells; and

d) injecting a therapeutically effective amount of said concentrated neural stem cells into one or more areas of a spinal cord of the subject.

2. The method of claim 1, wherein the spinal cord injury is associated with paralysis.

3. The method of claim 1, wherein the spinal cord injury comprises neuronal injury resulting from reduced blood flow to the spinal cord.

4. The method of claim 3, wherein the reduced blood flow is caused by ischemia, thoracic/abdominal aorta surgery, anterior spinal artery syndrome, chronic spinal spondylosis, or trauma.

5. The method of claim 1, wherein the neural stem cells are multipotential.

6. The method of claim 1, wherein the neural stem cells are derived from human embryonic stem cells.

7. The method of claim 1, wherein the fetal spinal cord tissue is obtained from a post-mortem fetus having a gestational age of about 6.5 to about 20 weeks.

8. The method of claim 1, wherein the growth factor is selected from the group consisting of bFGF, EGF, TGF-alpha, aFGF and combinations thereof.

9. The method of claim 1, wherein the neural stem cells are expanded to confluence.

10. The method of claim 1, wherein the injected neural stem cells generate at least 1,000 GABA-producing neurons in said tissue.

11. The method of claim 1, wherein said concentrating step c) comprises concentrating the expanded population to a density of about 1,000 cells/μ

L to about 200,000 cells/μ

L.

12. The method of claim 1, wherein said concentrating step c) comprises concentrating the expanded population to a density of about 5,000 cells/μ

L to about 50,000 cells/μ

l.

13. The method of claim 1, wherein said injecting step d) comprises multiple injections of the concentrated neural stem cells.

14. The method of claim 1, wherein at least 1,000 cells from said concentrated neural stem cells are injected to the spinal cord of the subject.

15. The method of claim 1, wherein at least 20% of the concentrated neural stem cells differentiate into neurons in the spinal cord of the subject.

16. The method of claim 1, wherein said concentrated neural stem cells are injected into the ventral horn area of the spinal cord.

17. The method of claim 1, wherein said concentrated neural stem cells are injected into the lumbar spinal cord.

18. The method of claim 1, wherein 0.1 to 100 μ

L of said concentrated neural stem cells are injected into one or more areas of a spinal cord of the subject.