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Application of compound 28d in preparation of medicine for increasing expression quantity of gamma-globin

A 28d, compound technology, applied in the field of application of compound 28d in the preparation of drugs that increase the expression of γ-globin

Active Publication Date: 2020-08-04
CHINA AUSTRALIA INST OF TRANSLATIONAL MEDICINE CO LTD NANJING CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these treatments can only relatively alleviate the disease. Studies have shown that drug induction and reactivation of the closed expression of γ-globin in adults can significantly reduce or treat the clinical symptoms of the above-mentioned anemia patients

Method used

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  • Application of compound 28d in preparation of medicine for increasing expression quantity of gamma-globin
  • Application of compound 28d in preparation of medicine for increasing expression quantity of gamma-globin
  • Application of compound 28d in preparation of medicine for increasing expression quantity of gamma-globin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Compound 28d increases the expression of γ-globin in K562 cells

[0020] experimental method

[0021] 1) Cell culture

[0022] Human erythroid K562 cells were cultured at 37°C in RPMI1640 medium, 10% (V / V) calf serum, 10 5 U / L Penicillin, 100mg / L Streptomycin, 5% CO 2 . K562 cells were treated with a final concentration of 5 μM for 28 days, and the control group was treated with DMSO (volume ratio <0.01%), and the cells were collected 48 hours later.

[0023] HUDEP-2 cells were treated with 10 μM dexamethasone, 1 μg / ml Dox, 50 ng / ml SCF, 3 U / ml EPO, 2% (V / V) double antibody, 1% (V / V) L-glutamine Incubate in SFEM medium at 37°C, add 5 μM for 28 days, and add DMSO (volume ratio <0.01%) for the control group, and collect cells after 48 hours.

[0024] ) RNA extraction, real-time PCR analysis

[0025] Add 1ml Trizol reagent (Invitrogen, USA) to an appropriate amount of cells. After the cells are fully broken and lysed, add 200 μl chloroform and centrifuge at...

Embodiment 2

[0030] Example 2 Compound 28d increases the expression of γ-globin in HUDEP-2 cells

[0031] experimental method:

[0032] 1) Cell culture: same as Example 1.

[0033] 2) RNA extraction, real-time PCR analysis: same as Example 1.

[0034] 3) Western blot experiment: same as Example 1.

[0035] 4) Flow cytometry analysis

[0036] After HUDEP-2 cells were treated with drugs (28d, 5μM) for 48 hours (2.5×10 7 ) were collected by centrifugation, fully washed with PBS and fixed with glutaraldehyde at a final concentration of 0.05% (V / V). Then wash 3 times with PBS containing 0.1% (W / V, g / 100ml) BSA and permeabilize with Triton X-100 with a final concentration of 0.1% (V / V), wash the permeabilized cells with PBS for 3 times and incubated with fetal hemoglobin antibody for 20 minutes, and finally measured by flow cytometry.

[0037] see results figure 2 .

[0038] 28d can increase the expression of γ-globin in HUDEP-2 cells: Compared with the control, the analysis found that th...

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Abstract

The invention belongs to the technical field of biological medicines, and discloses application of a compound 28d in preparation of a medicine for increasing the expression quantity of gamma-globin, and application in preparation of a medicine for treating beta-thalassemia or sickle-cell anemia.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to the new application of compound 28d (1,2-{N1-[2(2-chlorophenyl)thiobenzyl], N1-methylene}hydrazine), especially Application of compound 28d in the preparation of a drug for increasing the expression of γ-globin, and in the preparation of a drug for treating β-thalassemia or sickle cell anemia. Background technique [0002] Human embryonic, fetal, and adult hemoglobin is temporally expressed during red blood cell development and ontogeny. Different globins in the human body are mainly expressed by two gene clusters - the α gene cluster on chromosome 16 (containing the embryonic ζ gene and the adult α1 and α2 genes) and the b gene cluster on chromosome 11 (containing fetus G gamma and A γ, adult ε and β genes). Embryonic hemoglobin (ζ 2 ε 2 ) converts to fetal hemoglobin (HbFα 2 gamma 2 ), gradually replaced by adult hemoglobin (HbAα 2 beta 2 ) replaced. After two year...

Claims

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Application Information

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IPC IPC(8): A61K31/145A61P7/06
CPCA61K31/145A61P7/06
Inventor 赵权罗成王莹孔令东张明
Owner CHINA AUSTRALIA INST OF TRANSLATIONAL MEDICINE CO LTD NANJING CHINA
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