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

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Problems solved by technology

However, these treatments can only relatively alleviate the disease. Studies have shown that drug induction and reactivation of the c

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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PUM

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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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