Application of anti-CD47 monoclonal antibody to medicines used for treating acute lymphocytic leukemia cells with amycin resistance

A technology of acute lymphocytes and leukemia cells, which is applied in the field of medicine and can solve problems that have not been reported

Pending Publication Date: 2021-04-16
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Currently, anti-CD47 monoclonal antibodies have not been reported in the treatment of ADM-resistant hematological malignancies

Method used

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  • Application of anti-CD47 monoclonal antibody to medicines used for treating acute lymphocytic leukemia cells with amycin resistance
  • Application of anti-CD47 monoclonal antibody to medicines used for treating acute lymphocytic leukemia cells with amycin resistance
  • Application of anti-CD47 monoclonal antibody to medicines used for treating acute lymphocytic leukemia cells with amycin resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: In vitro detection of antibody-dependent cell-mediated phagocytosis (ADCP)

[0016] PBMC-derived macrophages were prepared from healthy donors, incubated with trypsin / EDTA (Gibco) for 3–5 min and scraped gently to obtain macrophages. 5.5×10 4 macrophages per well in a 24-well plate. Tumor cells were labeled with the standard dye 3 μM carboxyfluorescent succinimidyl ester (CFSE) according to standard protocols. After the macrophages were incubated in serum-free IMDM for 2-4 hours, 2.5×10 5 CFSE-labeled live tumor cells. Add 10-15 μg / mL anti-CD47 monoclonal antibody (Bio X Cell, USA), and incubate at 37°C for 2-4 hours. After incubation, the microwells were thoroughly washed 3-5 times with PBS and subsequently imaged with a fluorescence microscope. The phagocytosis index was calculated according to the number of effector cells (macrophages) that phagocytized CFSE-positive tumor cells. Analysis of results by fluorescence microscopy showed that anti-CD47 mo...

Embodiment 2

[0017] Example 2: In vivo effect of anti-CD47 monoclonal antibody against doxorubicin-resistant acute lymphoblastic leukemia systemic mouse model

[0018] 3-4 weeks old NOD / SCID mice were selected, and the doxorubicin-resistant acute lymphoblastic leukemia cell line Nalm6-Luc / ADR was used to establish a systemic B-ALL drug-resistant mouse model. First, a doxorubicin-resistant human B-ALL cell line Nalm6-Luc / ADR expressing luciferase in logarithmic growth phase was isolated. Count 3 x 10 4 Cells were injected into NOD / SCID mice through the tail vein. After the mouse model was established successfully, they were divided into PBS control group and anti-CD47 monoclonal antibody treatment group, with 10 mice in each group, and the dosage of anti-CD47 monoclonal antibody was 100ug / Only the control group was treated with PBS. Administration for 3 days, observation time 14 days. Mouse experiments were performed according to the guidelines for animal experiments of Fujian Medical U...

Embodiment 3

[0020] Example 3: In vivo effect of anti-CD47 monoclonal antibody against doxorubicin-resistant acute lymphoblastic leukemia subcutaneous xenograft tumor mouse model

[0021] NOD / SCID mice aged 3-4 weeks were selected, and the B-ALL drug-resistant mouse model with subcutaneous xenograft tumor was established with the doxorubicin-resistant acute lymphoblastic leukemia cell line Nalm6-Luc / ADR. Isolate the doxorubicin-resistant human B-ALL cell line Nalm6-Luc / ADR expressing luciferase in the logarithmic growth phase, counting 5×10 6 Cells were subcutaneously injected into the dorsal side of the right hind leg. After the successful construction of the subcutaneous transplanted tumor mouse model, they were divided into PBS negative control group and anti-CD47 monoclonal antibody treatment group, with 5 mice in each group, and the dosage of anti-CD47 monoclonal antibody was 100ug. per mouse, administered for 3 days, observed tumor growth, and recorded by IVIS bioluminescent imaging ...

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Abstract

The invention discloses application of an anti-CD47 monoclonal antibody to acute lymphocytic leukemia cells with amycin resistance, and relates to the activity of the anti-CD47 monoclonal antibody in treating acute lymphocytic leukemia with amycin resistance. According to the invention, PBMC-derived macrophages are prepared from a healthy donor; anti-CD47 monoclonal antibody-mediated phagocytosis is detected by antibody-dependent cell-mediated phagocytosis (ADCP); and by constructing an amycin-resistant acute lymphocytic leukemia cell strain Nalm6-Lu / ADR subcutaneous transplantation tumor mouse model and a systemic xenograft mouse model, the anti-CD47 monoclonal antibody acts on a model mouse, and the effect of the anti-CD47 monoclonal antibody on in-vivo treatment of amycin-resistant acute lymphocytic leukemia is observed.

Description

technical field [0001] The novel application relates to the field of medicine, and is the application of an anti-CD47 monoclonal antibody against doxorubicin-resistant acute lymphoblastic leukemia. Background technique [0002] Acute lymphoblastic leukemia (ALL) is a group of highly heterogeneous blood system tumors in which the proliferation and differentiation of malignant clones of hematopoietic stem / progenitor cells are blocked, accounting for about 20% of acute leukemia in adults and 80-85% of acute leukemia in children %. Despite the deepening research on the biological characteristics of leukemia, the current progress in the treatment of ALL is still limited. Although the first remission rate of induction therapy can reach 89%-90%, the disease-free survival rate of 3-5 years is only 10%-30%, and there are still some patients who do not respond to induction chemotherapy or are in complete remission (CR). After relapse, it eventually evolves into relapsed or refractor...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P35/02
Inventor 胡建达
Owner FUJIAN MEDICAL UNIV UNION HOSPITAL
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