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Combined drug for treating leukemia and its application in treating leukemia

A leukemia and chemotherapy drug technology, applied in the field of medicine, can solve the problems of easy relapse, poor chemotherapy sensitivity of leukemia, difficult clinical treatment of leukemia, etc., and achieve the effect of prolonging survival and obvious inhibitory effect

Active Publication Date: 2019-09-03
SHANGHAI TISSUEBANK BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chemotherapy sensitivity of refractory leukemia is extremely poor, and it is easy to relapse, which is a difficult problem in the clinical treatment of leukemia

Method used

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  • Combined drug for treating leukemia and its application in treating leukemia
  • Combined drug for treating leukemia and its application in treating leukemia
  • Combined drug for treating leukemia and its application in treating leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Targeted knockdown of histone deacetylase 3 can enhance the apoptosis-inducing effect of doxorubicin on K562 doxorubicin-resistant cells.

[0035] In this example, small hairpin RNA (shRNA, whose carrier is pLVX-shRNA2, purchased from Clontech Company) was used to knock down K562 doxorubicin-resistant cells (cells were purchased from the Institute of Hematology, Hematology Hospital, Chinese Academy of Medical Sciences) (hereinafter referred to as KA The expression of histone deacetylase 3 (HDAC3) in cells), and the knockdown efficiency was verified by western blotting.

[0036] Take 2×10 5 Cells were resuspended at a density of / ml, treated in different ways, and then collected at 24 hours, 48 ​​hours and 72 hours, and the proportion of positive cells (ie, apoptotic cells) was detected by Annexin V-APC single staining flow cytometry.

[0037] The treatment groups were KA cell group, KA cell HDAC3 knockdown group, KA cell HDAC3 knockdown and 0.5 μg / ml doxorub...

Embodiment 2

[0041]Example 2: Targeted inhibition of histone deacetylase 3 can enhance the apoptosis-inducing effect of cytarabine on THP-1 cells.

[0042] In this embodiment, 2×10 5 Resuspend THP-1 cells at a density of / ml (the cells were purchased from the Cell Bank of the Chinese Academy of Sciences), adopt different treatment methods, and then collect the cells at 24 hours and 48 hours, and use the Annexin V-FITC / PI double staining method to detect positive cells (i.e. the proportion of apoptotic cells).

[0043] Each treatment group is control group, cytarabine (hereinafter referred to as Ara-c) treatment group, RGFP966 (HDAC3 specific inhibitor) treatment group, Ara-c and RGFP966 combined treatment group. The concentration of Ara-c is 5 μM, and the concentration of RGFP is 2 μM.

[0044] The Annexin V-FITC / PI staining protocol is as follows: (1) collect single cell suspension, centrifuge and wash the cells twice with ice-cold PBS; (2) take 1×10 5 Cells were resuspended in 100 μl ...

Embodiment 3

[0047] Example 3: Targeted knockdown of histone deacetylase 3 can increase the sensitivity of KA cells to doxorubicin.

[0048] In this example, after KA cells were infected with control virus or HDAC3-specific knockdown virus, the IC50 of doxorubicin on each cell was detected using CCK-8 kit (the kit was purchased from Dongren Chemical Technology (Shanghai) Co., Ltd.) value (half inhibition rate).

[0049] The following is the principle of CCK-8 detection of cell survival and proliferation: CCK-8 reagent contains WST-8, which can be reduced to highly water-soluble yellow formazan product (Formazan) under the action of mitochondrial dehydrogenase. The amount of formazan product produced is directly proportional to the number of viable cells. Then, the absorbance value (A) at a wavelength of 450nm is detected by an enzyme-linked immunoassay instrument, which can reflect the number of living cells.

[0050] Take cells in the logarithmic growth phase, adjust the concentration t...

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Abstract

The invention belongs to the field of medicines, and discloses a combined medicine for treating leukemia and application thereof to treatment of the leukemia. The combined medicine combines a histone deacetylase 3 inhibitor and chemotherapeutic medicines commonly used by clinical acute myeloid leukemia, so that complete remission rates of new leukemia and refractory relapsed leukemia can be significantly increased, and sensibility of refractory relapsed leukemia cells to the chemotherapeutic medicines can be reversed more effectively.

Description

technical field [0001] The invention relates to the field of medicines, in particular to a combination medicine for treating leukemia and its application in treating leukemia. Background technique [0002] Acute myeloid leukemia is a common hematological malignancy, characterized by sudden onset, dangerous clinical manifestations, and poor prognosis, and poses a great threat to life and health. [0003] In current clinical practice, chemotherapy regimens based on cytarabine combined with an anthracycline antibiotic are often used for acute myeloid leukemia. Cytarabine is a pyrimidine anti-metabolite drug that inhibits DNA synthesis in cells and causes DNA damage, thereby causing tumor cell death. Anthracyclines are type II DNA topoisomerase inhibitors that can cause double-strand breaks in DNA, resulting in DNA damage. In addition, there are some improved chemotherapy regimens on this basis. But for now, although about 60% of acute myeloid leukemia patients can achieve cl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/06A61K31/704A61K31/7068A61K31/19A61P35/02
Inventor 龙俊胡炯李军民郑仲征
Owner SHANGHAI TISSUEBANK BIOTECH
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