Peptide medicament series for accelerating medulla hematopoiesis cell proliferation
A technology of bone marrow hematopoietic cells and drugs, applied in the field of biotechnology and medicine, can solve problems such as limited sources, achieve normal cell structure and promote cell proliferation
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[0030] The preparation method of the short peptide adopts a chemical synthesis method, that is, a very mature solid-phase peptide synthesis method well known in the art, and either the Boc method or the Fmoc method can be used. The specific method is to couple the protected amino acids to the inert solid phase carrier one by one, and then use strong acid to cleave the peptide chain from the carrier, and remove the side chain protection at the same time.
[0031]The amino acid main sequence of the short peptide is a main sequence with fixed amino acid positions, and the corresponding amino acids are selected and arranged at the interval positions. For example, there are two kinds of Ala or Ile amino acids at the J position; these two kinds of amino acids can be placed in a single place At the position of J, no matter which one is selected, the short peptide still has its original biological activity, that is, it promotes the proliferation of bone marrow hematopoietic cells, and ...
Embodiment 1
[0038] Promoting the proliferation of bone marrow nucleated cells in patients with aplastic anemia (synthesizing the short peptide derivatives involved in the present invention, modifying serine with a phosphate group: Ile-Val-Thr-Serp-Leu-Thr-Lys)
[0039] Bone marrow cells from human aplastic anemia patients were isolated from mononuclear cells by lymphocyte separation medium, and planted into 96-well cell plates. 5 Cell density per well was planted, and the short peptides involved in the present invention were added to the cell culture plate according to different concentrations, and an equal volume of PBS was used as the control group. The experimental results showed that the bone marrow nucleated cells were under the action of short peptide drugs. , the rate of promoting proliferation reached 121% in 3 days.
Embodiment 2
[0041] Morphological examination after promoting bone marrow nucleated cell proliferation (synthesis of short peptide fragments involved in the present invention, truncating 4 or 5 amino acids in the sequence: Ile-Val-Thr-Asp-Leu or Thr-Asp-Leu-Thr- Lys)
[0042] Bone marrow nucleated cells were cultured with DMEM medium containing 10% fetal calf serum, and the culture medium was 10 6 After the cell density per well was planted into a 24-well cell plate, the short peptides involved in the present invention were immediately added to the cell culture plate at different concentrations, and an equal volume of PBS was used as the control group. After 24 hours, the cells were centrifuged and removed. Supernatant, pre-fixed cell pellets with 3% glutaraldehyde-1.5% paraformaldehyde for 1 day (4°C), post-fixed with 1% osmic acid-1.5% potassium ferrocyanide for 1.5h, rinsed with PBS; 70% alcohol Block staining with saturated uranyl acetate dye solution, alcohol-acetone gradient dehydra...
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