Use of adipose tissue-derived stromal stem cells in treating fistula

• US 10,548,924 B2
• Filed: 09/03/2013
• Issued: 02/04/2020
• Est. Priority Date: 08/25/2004
• Status: Active Grant

• Current Assignee: Takeda Pharmaceutical Company Limited, Universidad Autónoma de Madrid
• Original Assignee: Tigenix SAU (Takeda Pharmaceutical Company Limited)
• Inventors: Miguel, Maria Gema Fernandez, De La Pena, Manuel Angel Gonzalez, Castro, Rosa Ana Garcia, Arranz, Mariano Garcia, Garcia-Olmo, Damian
• Primary Examiner(s): Leavitt, Maria G
• Application Number:
  US14/017,152
• Publication Number:
  US 20140072539A1
• Time in Patent Office: 2,345 Days
• Field of Search: None
• CPC Class Codes: A61B 17/1659 Surgical rasps, files, planes, or scrapers
• CPC Class Codes: A61B 17/32 Surgical cutting instruments A61B18/042 takes precedence; suture cutters A61B17/0467; instruments for ligaturing or cutting A61B17/128; instruments for rupturing the amniotic membrane A61B17/4208; specially adapted knives for eye surgery A61F9/0133
• CPC Class Codes: A61B 2017/320008 Scrapers
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 24/0015 Medicaments; Biocides
• CPC Class Codes: A61L 24/106 Fibrin; Fibrinogen
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3834 Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3869 Epithelial tissues other than skin
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/12 Antidiarrhoeals
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/02 Nasal agents, e.g. decongestants
• CPC Class Codes: A61P 13/10 : of the bladder
• CPC Class Codes: A61P 15/00 : Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/02 : for disorders of the vagina
• CPC Class Codes: A61P 17/00 : Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 : for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/16 : Otologicals
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/14 : Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• CPC Class Codes: C12N 2509/00 : Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 2533/56 : Fibrin; Thrombin
• CPC Class Codes: C12N 5/0667 : Adipose-derived stem cells [ADSC]; Adipose stromal stem cells

Abstract

Provided herein are novel methods and compositions utilizing adipose tissue-derived stromal stem cells for treating fistulae.

Claims

1. A method of preparing a pharmaceutical composition, comprising thawing a cryopreserved composition and formulating the thawed composition with human serum albumin (HSA) and Dulbecco'"'"'s Modified Eagle'"'"'s Medium (DMEM) to produce the pharmaceutical composition, wherein the cryopreserved composition comprises an isolated cell population comprising adipose tissue-derived stromal stem cells, wherein the isolated cell population was ex vivo generated by passaging adherent cells obtained from adipose tissue at least four times so that fewer than 10% of the adipose tissue derived stromal cells in the cryopreserved composition express the CD34 marker as measured by flow cytometry and at least 50% of the adipose-tissue derived stromal cells in the cryopreserved composition express the CD90 marker as measured by flow cytometry, wherein the concentration of the adipose tissue-derived stromal stem cells in the cryopreserved composition is at least about 5×

10⁶ cells/mL.

2. The method of claim 1 wherein at least 85% of the adipose tissue-derived stromal stem cells in the cryopreserved composition express the CD44 marker and more than 50% of the adipose tissue-derived stromal stem cells express the CD29 and CD105 markers as measured by flow cytometry.

3. The method of claim 1 wherein at least 95% of the adipose tissue-derived stromal stem cells in the cryopreserved composition express the CD44 marker as measured by flow cytometry.

4. The method of claim 1 wherein at least 99% of the adipose tissue-derived stromal stem cells in the cryopreserved composition express the CD44 marker as measured by flow cytometry.

5. The method of claim 1 wherein the adherent cells obtained from adipose tissue were passaged at least six times.

6. The method of claim 1, wherein the cryopreserved composition further comprises HSA.

10. The method of claim 1, wherein the pharmaceutical composition further comprises an adhesive.

11. The method of claim 10, wherein the adhesive comprises a fibrin glue or gel.

12. The method of claim 1, wherein the pharmaceutical composition further comprises a therapeutic agent.

13. The method of claim 12, wherein the therapeutic agent comprises an anti-inflammatory agent, an immunosuppressive agent, a biological agent, an antibiotic, an antidiarrheal agent or a combination thereof.

7. A method of preparing a pharmaceutical composition, comprising formulating an isolated cell population with HSA and DMEM to produce the pharmaceutical composition, wherein the isolated cell population comprises adipose tissue-derived stromal stem cells, wherein the isolated cell population was ex vivo generated by and passaging adherent cells from adipose tissue at least four times so that fewer than 10% of the adipose tissue-derived stromal stem cells in the pharmaceutical composition express the CD34 marker as measured by flow cytometry and at least 50% of the adipose tissue derived stromal stem cells in the pharmaceutical composition express the CD90 marker as measured by flow cytometry, wherein the concentration of the adipose tissue-derived stromal stem cells in the pharmaceutical composition is at least about 5×

10⁶ cells/mL.

8. The method of claim 7, further comprising aliquoting the pharmaceutical composition into one or more doses of at least about 10×

10⁶ cells/dose.

9. The method of claim 7, further comprising aliquoting the pharmaceutical composition into one or more doses of at least about 20×

10⁶ cells/dose.

14. The method of claim 7 wherein the adherent cells obtained from adipose tissue were passaged at least six times.