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GM-CSF targeted medicine liposome for resisting acute myelogenous leukemia and preparation method thereof

An acute myeloid and liposome technology, applied in liposome delivery, antineoplastic drugs, drug combinations, etc., can solve the problems of less than 15% cure rate, expensive monoclonal antibodies, and can not be reused, etc., to achieve the cost of treatment Reduced, low cost, less toxic and side effects

Inactive Publication Date: 2008-06-04
THE SECOND AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Acute myeloid leukemia is usually treated with chemotherapy. The standard DA chemotherapy regimen can make 60%-80% of the initial patients achieve remission, but the cure rate of chemotherapy alone is less than 15%
However, monoclonal antibodies are very expensive, and if mouse antibodies are used, they will easily lead to the production of antibodies and cannot be reused

Method used

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  • GM-CSF targeted medicine liposome for resisting acute myelogenous leukemia and preparation method thereof
  • GM-CSF targeted medicine liposome for resisting acute myelogenous leukemia and preparation method thereof
  • GM-CSF targeted medicine liposome for resisting acute myelogenous leukemia and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0050] Preparation, purification and identification of embodiment 1DHAQ-GM-CSF targeted drug liposome

[0051] 1. Preparation of DHAQ liposomes

[0052] DHAQ liposomes were prepared by reverse evaporation method (see literature Szoke F. Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse-phase evaporation [J]. Proc Natl Acad Sci USA, 1978, 75 (9): 4194) .

[0053] Take 10mg of DHAQ raw material, dissolve it in 20ml of distilled water for later use, take 2g of lecithin (Pc) and 0.4g of cholesterol (Cho) (Pc:Cho=1:0.2, W / W), add 60ml of ether, stir until completely dissolved, The DHAQ solution was slowly added to the ether solution of Pc and Cho (W Pc :W DHAQ =200:1, organic phase:aqueous phase=3:1), stirred while adding, and then placed in an ultrasonic oscillator for about 30 minutes to vibrate until the aqueous phase and organic phase were mixed evenly, and a blue emulsion suspension was formed. The emulsion was evaporated ...

Embodiment 2

[0090] Embodiment 2. Cytotoxicity experiment

[0091] Using the MTT method. Take HL60 cells cultured for 1 week without drugs, and make 5×10 4 / ml of cell suspension, inoculated in 96-well plate, try to make the cells into single cells when preparing the cell suspension, shake the cell suspension gently when inoculating, so that the number of cells added to each well is consistent, Add 200 μl of cell suspension (1×10 4 cells / well).

[0092] The experiments were divided into four groups:

[0093] ①DHAQ-GM-CSF targeted drug liposome group: Add targeted drugs with concentrations (according to DHAQ content) of 0.0025 μg / ml, 0.025 μg / ml, 0.25 μg / ml, 2.5 μg / ml, and 25 μg / ml in sequence Liposome solution, 20 μl / well.

[0094] ②DHAQ liposome group: Add liposome solutions with concentrations (according to the concentration of DHAQ) of 0.0025 μg / ml, 0.025 μg / ml, 0.25 μg / ml, 2.5 μg / ml, and 25 μg / ml in sequence, 20 μl / well.

[0095] ③DHAQ group: DHAQ solutions with concentrations of...

Embodiment 3

[0112] Example 3 DHAQ-GM-CSF targeting drug liposome GM-CSF guiding effect verification experiment

[0113] In the present invention, we carried out the verification experiment of GM-CSF orientation while carrying out the MTT experiment. This experiment shows that GM-CSF on DHAQ-GM-CSF targeting liposome can play a guiding role, and its guiding effect is effected through ligand-receptor binding. If a saturated amount of GM-CSF is added in advance to occupy HL60 The receptors on the cell surface, then the specific killing effect of the DHAQ-GM-CSF targeted drug liposome added later will be blocked, leaving only the non-specific killing effect of the liposome. Adopt MTT method to carry out simultaneously with embodiment 2.

[0114] Method is with embodiment 2.

[0115] The experiment was divided into 5 dosage groups:

[0116] Each group first added 7.5ng saturated amount of GM-CSF (containing 3×10 11 GM-CSF molecules, 10 of the cell binding sites per well 4 times), placed i...

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Abstract

The invention discloses a GM-CSF target-direction drug liposome and preparing method of anti acute leukemia myelomatosis, which is composed of cytokine guiding matter with initiative target-tropism connecting to the surface of liposome and anti acute leukemia myelomatosis drug in liposome. The external model MTT of the invention experiments and checks the toxic effect on the cell of acute leukemia myelomatosis cell strain HL60 and the effect on cell death. The method improves the curative effect, which reduces the cost.

Description

technical field [0001] The invention relates to a drug for treating acute myeloid leukemia, specifically, a targeting agent formed by linking active targeting guide GM-CSF on the surface of a liposome encapsulated with an anti-acute myeloid leukemia drug. Drug liposome and its preparation method. Background technique [0002] Acute leukemia is a malignant tumor of the hematopoietic system, with an incidence of about 3 / 100,000 residents, ranking first among adolescent malignant tumors. The cause of the disease is not very clear at present, and it may be related to viruses, physical and chemical carcinogens. Acute leukemia often leads to normal hematopoietic dysfunction and leukemic cell infiltration symptoms, clinical manifestations such as severe infection, bleeding, anemia, etc., and the median survival period without treatment is less than three months, posing a great threat to life and property. Acute leukemia is divided into two main types: acute myeloid leukemia and a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/127A61K45/00A61K47/48A61P35/00A61K47/62
Inventor 娄世锋张颖陈林陈姝翁春岚彭薇周慷邓建川罗云
Owner THE SECOND AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIV
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