Compositions for the mobilization, homing, expansion and differentiation of stem cells and methods of using the same

• US 9,808,454 B2
• Filed: 03/17/2014
• Issued: 11/07/2017
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Ampio Pharmaceuticals, Inc.
• Original Assignee: Ampio Pharmaceuticals, Inc.
• Inventors: Bar-Or, David, Thomas, Greg
• Primary Examiner(s): Ton, Thaian N
• Assistant Examiner(s): Montanari, David A
• Application Number:
  US14/216,430
• Publication Number:
  US 20140286913A1
• Time in Patent Office: 1,331 Days
• Field of Search: 424 937
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/20 having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
• CPC Class Codes: A61K 31/405 Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
• CPC Class Codes: A61K 31/495 having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazinesA61K31/48 takes precedence ; with three nitrogen atoms A61K31/53
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

The invention provides compositions that increase the mobilization, homing, expansion, and/or differentiation of stem cells and methods of using the same for the treatment of mammals.

Claims

1. A method of expanding bone marrow derived mesenchymal stem cells, comprising

(a) contacting the mesenchymal stem cells with DA-DKP, wherein the DA-DKP is in a composition comprising a less than 5000 molecular weight fraction of human serum albumin, and

(b) culturing the stem cells under conditions suitable to facilitate the expansion of the mesenchymal stem cells.

5. The method of claim 1, wherein the DA-DKP composition further comprises N-acetyl tryptophan, caprylate and/or caprylic acid.

6. The method of claim 1, wherein the DA-DKP composition comprises a fraction of human serum albumin, wherein substantially all of the albumin has been removed from the fraction.

7. The method of claim 1, wherein the low molecular weight fraction of human serum albumin is produced by filtration.

8. The method of claim 1, wherein the step of contacting the mesenchymal stem cells with DA-DKP has an effect selected from the group consisting of increasing production of a protein selected from the group consisting of CXCR4, MMP14, MMP13, aggrecan, SDF1, collagen 2A1 and combinations thereof and decreasing production of CXCL12.

9. The method of claim 1, wherein the step of culturing includes the addition of one or more components selected from the group consisting of thrombopoietin (TPO), stem cell factor (SCF), IL-1, IL-3, IL-7, flt-3 ligand (flt-3L), G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, IGF, EGF, NGF, LIF, PDGF, bone morphogenic proteins (BMP), activin-A, VEGF, forskolin, and glucocorticords.

2. A method of providing mammalian bone marrow derived mesenchymal stem cells to a mammal, comprising:

(a) contacting the mesenchymal stem cells with DA-DKP, wherein the DA-DKP is in a composition comprising a less than 5000 molecular weight fraction of human serum albumin,

(b) cultivating the mesenchymal stem cells under conditions suitable to facilitate the expansion of the cells; and

(c) introducing the expanded cells into a mammal.

10. The method of claim 2, wherein the DA-DKP composition further comprises N-acetyl tryptophan, caprylate and/or caprylic acid.

11. The method of claim 2, wherein the DA-DKP composition comprises a fraction of human serum albumin, wherein substantially all of the albumin has been removed from the fraction.

12. The method of claim 2, wherein the low molecular weight fraction of human serum albumin is produced by filtration.

13. The method of claim 2, wherein the step of contacting the mesenchymal stem cells with DA-DKP has an effect selected from the group consisting of increasing production of a protein selected from the group consisting of CXCR4, MMP14, MMP13, aggrecan, SDF1, collagen 2A1 and combinations thereof and decreasing production of CXCL12.

14. The method of claim 2, wherein the step of cultivating includes the addition of one or more components selected from the group consisting of thrombopoietin (TPO), stem cell factor (SCF), IL-1, IL-3, IL-7, flt-3 ligand (flt-3L), G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, IGF, EGF, NGF, LIF, PDGF, bone morphogenic proteins (BMP), activin-A, VEGF, forskolin, and glucocorticords.

3. A method of causing bone marrow derived mesenchymal stem cells to differentiate into a particular type of cell comprising:

(a) contacting the mesenchymal stem cells with DA-DKP, wherein the DA-DKP is in a composition comprising a less than 5000 molecular weight fraction of human serum albumin;

(b) cultivating the mesenchymal stem cells under conditions suitable to facilitate the expansion of the stem cells; and

(c) adding one or more differentiation factors or changing culturing conditions to induce differentiation of the mesenchymal stem cells to form a different type of cell.

15. The method of claim 3, wherein the DA-DKP composition further comprises N-acetyl tryptophan, caprylate and/or caprylic acid.

16. The method of claim 3, wherein the DA-DKP composition comprises a fraction of human serum albumin, wherein substantially all of the albumin has been removed from the fraction.

17. The method of claim 3, wherein the low molecular weight fraction of human serum albumin is produced by filtration.

18. The method of claim 3, wherein the step of contacting the mesenchymal stem cells with DA-DKP has an effect selected from the group consisting of increasing production of a protein selected from the group consisting of CXCR4, MMP14, MMP13, aggrecan, SDF1, collagen 2A1 and combinations thereof and decreasing production of CXCL12.

19. The method of claim 3, wherein the step of cultivating includes the addition of one or more components selected from the group consisting of thrombopoietin (TPO), stem cell factor (SCF), IL-1, IL-3, IL-7, flt-3 ligand (flt-3L), G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, IGF, EGF, NGF, LIF, PDGF, bone morphogenic proteins (BMP), activin-A, VEGF, forskolin, and glucocorticords.

20. The method of claim 3, wherein the step of adding one or more differentiation factors or changing culturing conditions comprises a step selected from the group consisting of alteration in temperature, alteration in oxygen/carbon dioxide content, alteration in media turbidity, and addition of a differentiation factor selected from nutrients, enzyme inhibitors, enzyme stimulators, histone deacetylase activity inhibitors, DNA methyltransferase activity inhibitors, and enzyme GSK-3 inhibitors.

4. A method of providing differentiated cells, to a mammalian subject, comprising:

(a) contacting mammalian multipotent bone marrow derived mesenchymal stem cells with DA-DKP, wherein the DA-DKP is in a composition comprising a less than 5000 molecular weight fraction of human serum albumin;

(b) cultivating the mesenchymal stem cells under conditions suitable to facilitate the expansion of the mesenchymal stem cells;

(c) adding one or more differentiation factors or changing culturing conditions to induce differentiation of cells to form a different type of cell; and

(d) introducing the differentiated cells into the subject.

21. The method of claim 4, wherein the DA-DKP composition further comprises N-acetyl tryptophan, caprylate and/or caprylic acid.

22. The method of claim 4, wherein the DA-DKP composition comprises a fraction of human serum albumin, wherein substantially all of the albumin has been removed from the fraction.

23. The method of claim 4, wherein the low molecular weight fraction of human serum albumin is produced by filtration.

24. The method of claim 4, wherein the step of contacting the mesenchymal stem cells with DA-DKP has an effect selected from the group consisting of increasing production of a protein selected from the group consisting of CXCR4, MMP14, MMP13, aggrecan, SDF1, collagen 2A1 and combinations thereof and decreasing production of CXCL12.

25. The method of claim 4, wherein the step of cultivating includes the addition of one or more components selected from the group consisting of thrombopoietin (TPO), stem cell factor (SCF), IL-1, IL-3, IL-7, flt-3 ligand (flt-3L), G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, IGF, EGF, NGF, LIF, PDGF, bone morphogenic proteins (BMP), activin-A, VEGF, forskolin, and glucocorticords.

26. The method of claim 4, wherein the step of adding one or more differentiation factors or changing culturing conditions comprises a step selected from the group consisting of alteration in temperature, alteration in oxygen/carbon dioxide content, alteration in media turbidity, and addition of a differentiation factor selected from nutrients, enzyme inhibitors, enzyme stimulators, histone deacetylase activity inhibitors, DNA methyltransferase activity inhibitors, and enzyme GSK-3 inhibitors.