Pharmaceutical composition for treating T-cell acute lymphocytic leukemia

An acute lymphocyte and composition technology, applied in the field of biomedicine, can solve the problems of easy recurrence, heterogeneity and poor prognosis, limited effective diagnosis and treatment methods, etc., and achieve the effect of improving the effect.

Active Publication Date: 2021-09-28
WUHAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

T-ALL is a rapidly developing and aggressive hematological malignancy, with great heterogeneity, poor prognosis, and easy relapse clinical characteristics, and effective clinical diagnosis and treatment methods are very limited

Method used

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  • Pharmaceutical composition for treating T-cell acute lymphocytic leukemia
  • Pharmaceutical composition for treating T-cell acute lymphocytic leukemia
  • Pharmaceutical composition for treating T-cell acute lymphocytic leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the impact of gene knockout on T-ALL cell proliferation

[0030]1. Acquisition of siRNA: siRNA (5 nmol) of WTAP, METTL3, METTL14, and FTO, 2 pairs for each and a negative control, designed and synthesized by Ribobio;

[0031]

[0032]

[0033] It should be noted that shRNA, that is, small hairpin RNA or short hairpin RNA (shRNA) is an RNA sequence with a tight hairpin turn, which is often used for RNA interference to silence the expression of target genes . The shRNA is introduced into the cells using a vector, and the U6 promoter in the vector ensures that the shRNA is always expressed; this shRNA-loaded vector can be passed to daughter cells, so that gene silencing can be inherited. The hairpin structure of shRNA can be cut into siRNA by cellular machinery, and then siRNA binds to RNA-induced silencing complex (RISC), which can bind to target mRNAs and degrade them.

[0034] siRNA, small or short interfering RNA (small / short interfering RNA, siRN...

Embodiment 2

[0041] Total RNA extraction: Total RNA extraction was performed according to the instructions of Yisheng Total RNA Extraction Kit (MolPure Cell / Tissue Total RNA Kit, Yeasen) as follows:

[0042] (1) Sample pretreatment: collect the cell pellet by centrifugation, add 1mL lysate LB for every 5×106-1×107 cells, and pipette repeatedly.

[0043] (2) total RNA extraction:

[0044] Set the RNA adsorption column A1 into a 2mL collection tube for later use.

[0045] Add the above pretreatment mixture (and precipitate) into the RNA adsorption column A1, centrifuge at 12,000 rpm for 1 min, and discard the filtrate.

[0046] Add 500 μL protein-removing solution PL, centrifuge at 12,000 rpm for 45 sec, and discard the filtrate.

[0047] Put the RNA adsorption column A1 back into the collection tube, add 500 μL of washing solution W*, centrifuge at 12,000 rpm for 45 sec at room temperature, and discard the filtrate. Repeat step 4 again.

[0048] Put the RNA adsorption column A1 back int...

Embodiment 3

[0065] Example 3: CO-IP experiment to verify the interaction between the target protein WTAP and JMJD6

[0066] (1) Preparation of related reagents and related antibodies:

[0067] Sucrose-NP40 Lysis buffer:

[0068]

[0069] Add before use: 10mM NaF, 1mM NaVO 3 , 1mM DTT, 1mM PMSF, protease inhibitors

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Abstract

The invention discloses a pharmaceutical composition for treating T cell acute lymphocytic leukemia (T-ALL). The composition comprises an active component for inhibiting expression of WTAP and an active component for inhibiting expression of JMJD6. According to the pharmaceutical composition disclosed by the invention, the proliferation of T-ALL cells can be synergistically inhibited by simultaneously inhibiting the expression of WTAP and JMJD6.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a pharmaceutical composition for treating T-cell acute lymphoblastic leukemia and its application. Background technique [0002] Acute lymphoblastic leukemia (ALL) is a hematological malignancy in which lymphoid precursor cells are arrested in their immature stages of differentiation leading to malignant proliferation. ALL can arise in T-cell or B-cell precursors of lymphocytes and is therefore classified as T-ALL or B-ALL. Among them, T-cell acute lymphoblastic leukemia (T-ALL) accounts for 10%-15% of cases in children and 25% in adults. The heterogeneity of clinical characteristics and prognosis makes its treatment effect far inferior to that of B-lymphoblastic leukemia ( B-ALL), and the current treatment options for T-ALL are very limited. [0003] T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy that primarily affects children and adolescents. Mu...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K48/00A61K31/713A61P35/02
CPCA61K45/06A61K48/005A61K48/0008A61K31/713A61P35/02A61K2300/00
Inventor 祝海川
Owner WUHAN UNIV OF SCI & TECH
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