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Pharmaceutical composition for promoting hematopoiesis and preparation method thereof

A composition and drug technology, applied in the direction of drug combination, pharmaceutical formula, medical preparations containing active ingredients, etc., can solve the problems of slow onset, slow relief, low cure rate, etc., to achieve rapid onset, promote hematopoiesis, High cure rate effect

Inactive Publication Date: 2009-09-16
SOUTHWEST HOSPITAL OF CHONGQING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Zaofan Compound Pills with Saponin as the main drug are commonly used in the auxiliary treatment of certain hematopoietic disorders, and can also improve the hematopoietic function of radiation-damaged animals [Xiao Yang et al., "New Drugs and Clinical Pharmacology of Traditional Chinese Medicine", Vol. 15 , pages 387-389], but there are disadvantages of slow relief, slow onset of effect, and low cure rate

Method used

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  • Pharmaceutical composition for promoting hematopoiesis and preparation method thereof
  • Pharmaceutical composition for promoting hematopoiesis and preparation method thereof
  • Pharmaceutical composition for promoting hematopoiesis and preparation method thereof

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preparation example Construction

[0035] Another aspect of the present invention provides a preparation method of a hematopoietic pharmaceutical composition, which comprises: isolating and purifying hematopoietic stem cells and resuspending them in a pharmaceutically acceptable medium; dissolving soap alum in a pharmaceutically acceptable medium Medium; mix the above cell suspension with soap alum solution.

[0036] Another aspect of the present invention provides a preparation method of a hematopoietic pharmaceutical composition, which includes: isolating and purifying hematopoietic stem cells; introducing expression vectors carrying cytokines into the stem cells, and screening positive transfection cells; The cells are resuspended in a pharmaceutically acceptable medium; saponin is dissolved in the pharmaceutically acceptable medium; and the above cell suspension is mixed with the saponin solution.

[0037] In the present invention, the medium for resuspending cells and the medium for dissolving soap alum ma...

Embodiment 1

[0051] Separation and purification of umbilical cord blood stem cells to obtain CD34 + cell

[0052] The umbilical cord blood of normal full-term male neonates (provided by the Southwest Hospital Affiliated to the Third Military Medical University) was aseptically collected, anticoagulated with heparin, and the mononuclear cells were separated by Ficoll lymphocyte separation medium (density 1.077g / ml) to contain 10 Wash twice in IMDM with % fetal calf serum, resuspend in 300 μl of PBS solution containing 0.5% BSA and 2 mmol / L EDTA, and isolate CD34 from mononuclear cells by immunomagnetic bead sorting system (mini-MACS) + cell. The specific method is as follows: add 100 μl of blocking solution (3% goat serum) and anti-CD34 monoclonal antibody to the above cell suspension, mix well, and place at 4° C. for 15 minutes. After centrifugation and washing with 10 ml of the above-mentioned PBS buffer, 100 μl of goat anti-mouse IgG immunomagnetic beads were added and placed at 4°C fo...

Embodiment 2

[0056] Construction of IL-3, FL and co-expression of FL and IL-3 bicistronic eukaryotic vector

[0057] The eukaryotic expression vector pIRES2-EGFP was purchased from Clontech Company. Escherichia coli DH5α strain was preserved by our laboratory. The plasmid MXIL-3neo containing full-length mouse IL-3 cDNA (2.5kb) was donated by Professor Moroni of University of Basel, Switzerland; the plasmid pUMVC3-hFL containing full-length human FL cDNA (0.8kb) was purchased from Gene Therapy Center, University of Michigan, USA.

[0058] 1. Construction of pIRES2-EGFP-IL-3:

[0059] 1) Amplification of IL-3 target fragment: use primer design software Primer5.0 to design primers as follows:

[0060] Upstream: 5-GGAATTCCCTGTGGCTTCTTCA-3 (SEQ ID NO: 1) (containing EcoRI site);

[0061] Downstream: 5-TCCCCGCGGGGAAAACCCATTTGTTC-3 (SEQ ID NO: 2) (containing SacII site).

[0062]The PCR reaction was carried out using the MXIL-3neo plasmid as a template. The reaction conditions were: denatura...

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Abstract

The invention relates to a pharmaceutical composition for promoting hematopoiesis, which comprises hemopoietic stem cells and melanterite, wherein the preferred hemopoietic stem cells are cells which are imported with an expression vector with cell factor genes. The invention also relates to a method for preparing the pharmaceutical composition and application of the pharmaceutical composition in preparing a medicament for treating the damage state of hematopoietic function, wherein the hemopoietic stem cells and the melanterite have the function of synergistically promoting the hematopoiesis.

Description

technical field [0001] The invention relates to a pharmaceutical composition for promoting hematopoiesis and a method for preparing the pharmaceutical composition. The present invention also relates to the use of the pharmaceutical composition in the preparation of medicaments for treating impaired hematopoietic function. Background technique [0002] Hematopoiesis is an important part of human life activities. In adult animals, the hematopoietic tissue is located primarily in the bone marrow. Under physiological conditions, the hematopoietic process is a dynamic process in which hematopoietic stem cells (HSCs) proliferate, differentiate, and form various mature blood cells. A variety of common blood system diseases in clinical practice, such as aplastic anemia, multiple myeloma, myelodysplastic syndrome, etc., as well as bone marrow damage caused by radiotherapy and chemotherapy, will lead to inhibition or failure of bone marrow hematopoietic function, which will bring ha...

Claims

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Application Information

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IPC IPC(8): A61K35/14A61P7/06A61K33/26A61K35/28
Inventor 张勇杜宏伟郭朝华董世武
Owner SOUTHWEST HOSPITAL OF CHONGQING
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