PHARMACEUTICAL COMPOSITION FOR CANCER TREATMENT AND PHARMACEUTICAL PREPARATION FOR CANCER TREATMENT CONTAINING SAME COMPOSITION AS ACTIVE INGREDIENT

• US 20170007677A1
• Filed: 01/23/2015
• Published: 01/12/2017
• Est. Priority Date: 01/24/2014
• Status: Active Grant

• Current Assignee: Cysay Incorporated
• Original Assignee: Quarrymen Corporation
• Inventors: Ueda, Minoru
• Granted Patent: US 10,639,355 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• 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 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 38/1841 Transforming growth factor [TGF]
• CPC Class Codes: A61K 38/1866 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: A61K 38/195 Chemokines, e.g. RANTES
• CPC Class Codes: A61K 38/30 Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2 insulin-like growth factor binding protein A61K38/1754
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 2500/02 Atmosphere, e.g. low oxygen conditions
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2500/99 Serum-free medium
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/21 Chemokines, e.g. MIP-1, MIP-2, RANTES, MCP, PF-4
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/1361 from dental pulp or dental follicle stem cells
• CPC Class Codes: C12N 2510/04 Immortalised cells
• CPC Class Codes: C12N 5/0664 Dental pulp stem cells, Dental follicle stem cells
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The invention provides a novel anticancer therapeutic having a high effect against solid cancers, lacking the side effects of chemotherapeutics, and being unlikely to cause resistance. Provided is a pharmaceutical composition for cancer treatment prepared by a method having: a stem cell production step for making immortalized stem cells by introducing four types of genes into deciduous tooth dental pulp stem cells obtained from the dental pulp of mammals; and a condition medium preparation step for culturing the immortalized stem cells for a predetermined length of time in serum-free medium at 23-27° C. under conditions of low oxygen concentration at an oxygen concentration of from 0.5% to less than 20%, the pharmaceutical composition containing 1.5 times or more of insulin-like growth factor (IGF-1) and vascular endothelial growth factor (VEGF) than is contained in condition medium prepared when culturing at an oxygen concentration of 20% and otherwise identical conditions.

Claims

1. A pharmaceutical composition for cancer therapy prepared by a method comprising the steps of:

manufacturing an immortalized stem cell by transfecting 4 gene into dens deciduous dental pulp stem cell derived from mammal dental pulp; and

preparing a conditioned medium by culturing said immortalized stem cell in a serum free medium for predetermined period under the condition of hypoxic concentration of 0.5% or more but less than 20% of oxygen concentration between the temperature of 23 to 27°

C.;

wherein the conditioned medium comprises more than 1.5 times higher concentration of both insulin like growth factor 1 (IGF-1) and vascular endothelial cell growth factor (VEGF), compared to the conditioned medium prepared by culturing the cell on the same condition except the oxygen concentration is set to 20%.

2. The pharmaceutical composition for cancer therapy according to claim 1, wherein said predetermined period is 40 to 56 hours.

3. The pharmaceutical composition for cancer therapy according to claim 1, wherein said hypoxic concentration means that oxygen concentration is 5% or less of.

4. The pharmaceutical composition for cancer therapy according to claim 3, wherein said hypoxic concentration means the oxygen concentration is 1% or less.

5. The pharmaceutical composition for cancer therapy according to claim 4, wherein said 4 gene is selected from the group consisting hTERT, bmi-1, E6, E7, Oct3/4, Sox2, Klf4, c-Myc, and p16INK4a.

6. The pharmaceutical composition for cancer therapy according to claim 1 further comprises at least five times higher concentration of transforming growth factor β

(TGF-β

1) compared to the conditioned medium prepared by culturing the stem cell under the same condition except the oxygen concentration is set to 20%.

7. The pharmaceutical composition for cancer therapy according to claim 5 further comprises at least three times higher concentration of stromal cell-derived factor (SDF-1).

8. A pharmaceutical preparation for cancer therapy comprises the pharmaceutical composition according to claim 1 as active ingredient.

9. The pharmaceutical preparation for cancer therapy according to claim 8, wherein said cancer is solid tumor.

10. The pharmaceutical composition for cancer therapy according to claim 2, wherein said hypoxic concentration means that oxygen concentration is 5% or less of.

11. The pharmaceutical composition for cancer therapy according to claim 10, wherein said hypoxic concentration means the oxygen concentration is 1% or less.

12. The pharmaceutical composition for cancer therapy according to claim 11, wherein said 4 gene is selected from the group consisting hTERT, bmi-1, E6, E7, Oct3/4, Sox2, K1f4, c-Myc, and p16INK4a.

13. The pharmaceutical composition for cancer therapy according to claim 6 further comprises at least three times higher concentration of stromal cell-derived factor (SDF-1).

14. The pharmaceutical composition for cancer therapy according to claim 12 further comprises at least three times higher concentration of stromal cell-derived factor (SDF-1).

15. A pharmaceutical preparation for cancer therapy comprises the pharmaceutical composition according to claim 5 as active ingredient.

16. A pharmaceutical preparation for cancer therapy comprises the pharmaceutical composition according to claim 6 as active ingredient.

17. A pharmaceutical preparation for cancer therapy comprises the pharmaceutical composition according to claim 12 as active ingredient.

18. The pharmaceutical preparation for cancer therapy according to claim 15, wherein said cancer is solid tumor.

19. The pharmaceutical preparation for cancer therapy according to claim 16, wherein said cancer is solid tumor.

20. The pharmaceutical preparation for cancer therapy according to claim 17, wherein said cancer is solid tumor.