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Application of levistilide A in preparation of medicine for in-vitro amplification of human hematopoietic stem cells

A technology of angelica lactone and in vitro amplification, applied in the field of application of angelica lactone A in the preparation of drugs for in vitro expansion of human hematopoietic stem cells, can solve problems such as unreported, and achieve good self-renewal and self-renewal Promoted effect

Active Publication Date: 2021-05-28
NANKAI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are no reports at home and abroad about the use of angelica lactone A in the in vitro expansion of human hematopoietic stem cells

Method used

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  • Application of levistilide A in preparation of medicine for in-vitro amplification of human hematopoietic stem cells
  • Application of levistilide A in preparation of medicine for in-vitro amplification of human hematopoietic stem cells
  • Application of levistilide A in preparation of medicine for in-vitro amplification of human hematopoietic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A kind of small molecule compound Levistilide A (Levistilide A), its structure is:

[0034]

[0035] The physical and chemical identification of the small molecule compound angelicalide A, including:

[0036] Purity determined by high performance liquid chromatography (HPLC): 98.8%.

[0037] H NMR ( 1 H NMR) (400MHz, CDCl3) δ7.34(d, J=6.6Hz, 1H), 5.06(t, J=7.9Hz, 1H), 4.99(t, J=7.5Hz, 1H), 3.24(d, J=8.9Hz, 1H), 2.98(dd, J=6.5, 2.1Hz, 1H), 2.54(t, J=7.7Hz, 1H), 2.28(q, J=7.6Hz, 2H), 2.24–2.14( m,3H),2.13–1.98(m,2H),1.97–1.82(m,2H),1.63–1.40(m,6H),1.30(ddd,J=12.2,4.7,2.6Hz,1H),0.98– 0.87(m,6H).

[0038] Carbon NMR ( 13 C NMR) (100MHz, CDCl3) δ168.6, 165.0, 155.1, 150.6, 148.1, 142.2, 134.3, 126.7, 112.3, 108.7, 47.7, 41.7, 41.6, 38.4, 31.2, 29.1, 28.1, 27.6, 25.9, 22.4, 2 19.9, 14.1, 13.9.

[0039] High resolution mass spectrometry HRMS (ESI) determination of C 24 h 29 o 4 + [M+H] + :381.2060, found 381.2062.

[0040] A kind of small molecular compound neo...

Embodiment 2

[0047] Compound effect of embodiment 2 different concentrations

[0048] Freshly isolated human cord blood CD34 + Cells were resuspended in culture medium (Gibco Iscove’s Modified Dulbecco Medium+10% FBS+100ng / mL SCF+100ng / mL TPO+100ng / mL Flt3L+1%P / S) to a cell concentration of 5×10 4 Cells / mL, then spread in 96-well plate, each hole was 190 μL cell suspension, and then added 10 μL concentration gradient of 2.5, 5, 10, 20 and 40 μM Euangicalide A (L) and new cnidium Ester (N) was cultured, and 0.05% DMSO with the same content was used as a solvent control. After 7 days of culture, flow cytometric analysis was performed to detect the ratio of CD34+CD38- hematopoietic stem progenitor cells and relatively more primitive CD34+CD49f+ cells in CD34+ cells.

[0049] Result analysis: if figure 1 As shown, for the CD34+CD38- cell population, both compounds exhibited an approximate dose-dependent expansion effect, and for the CD34+CD49f+ cell population, Euangicalide A significantly e...

Embodiment 3

[0051] A method for in vitro expansion of human hematopoietic stem cells, comprising the steps of:

[0052] Freshly isolated human cord blood CD34 + Cells were resuspended in culture medium (Gibco Iscove’s Modified Dulbecco Medium+10% FBS+100ng / mL SCF+100ng / mL TPO+100ng / mL Flt3L+1%P / S) to a cell concentration of 5×10 4 Cells / mL, then spread in a 96-well plate, each well is 190 μL of cell suspension, and then add 10 μL of the small molecule compound of Euangicalide A diluted with the same culture medium, the final concentration of Euangicalide A 10 μM. at 37°C, 5% CO 2 Cultured in a constant temperature incubator for 7 days.

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Abstract

The invention relates to application of levistilide A in preparation of a medicine for in-vitro amplification of human hematopoietic stem cells, in particular to application, in the application, the concentration of the levistilide A in an amplification culture medium is 1-10 [mu]M, and more preferably, the concentration of the levistilide A in the amplification culture medium is 5-10 [mu]M. The levistilactone A and the structure thereof have a good promotion effect on self-renewal of human hematopoietic stem cells, and a new method can be provided for clinical treatment in the future.

Description

technical field [0001] The present invention relates to a new application of angelica lactone A, in particular, the present invention relates to the application of angelica lactone A in the preparation of medicines for in vitro expansion of human hematopoietic stem cells. Background technique [0002] The regenerative potential of hematopoietic stem cells (HSCs) makes it play an important role in the treatment of leukemia, the reconstruction of hematopoietic function in cancer patients after chemotherapy, and the gene therapy of genetic diseases and tumors. Umbilical cord blood is rich in hematopoietic stem cells, and has the advantages of low immunogenicity, safe donor, wide range of sources, and easy collection. Therefore, umbilical cord blood transplantation has become an excellent alternative source for bone marrow and peripheral blood hematopoietic stem cell transplantation. [0003] However, the proportion of HSCs in umbilical cord blood is only about 0.1% to 1%. Due t...

Claims

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Application Information

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IPC IPC(8): C12N5/0789
CPCC12N5/0647C12N2501/999
Inventor 张泉高瀛岱李迎辉杨铭何媚丁亚辉张文姗徐惠王超群
Owner NANKAI UNIV
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