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Folic acid modified liposome complex and preparation method and use thereof

A liposome complex and liposome technology, applied in the field of medicine, can solve the problem that the immunosuppressive effect of the tumor microenvironment cannot be ignored.

Active Publication Date: 2019-06-14
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most current targeted cancer therapies focus on killing tumor cells, but the immunosuppressive effect of the tumor microenvironment cannot be ignored

Method used

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  • Folic acid modified liposome complex and preparation method and use thereof
  • Folic acid modified liposome complex and preparation method and use thereof
  • Folic acid modified liposome complex and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Tissue Microarray Analysis

[0042] 1. Experimental method

[0043] Prior to data extraction and tissue microarray (TMA) construction, approval for the study was obtained from the Ethics Committee of Shanghai Xinchao Biotechnology Co., Ltd. In the present experimental study, 184 patients diagnosed with primary lung squamous cell carcinoma and lung adenocarcinoma between 2004 and 2009 were selected. Extract clinical and pathological data from surgical pathology and medical records. Histological designations follow the World Health Organization classification, and grading and staging are based on the 8th edition of the American Joint Committee on Cancer (AJCC).

[0044]Primary tumors from 184 patients with pathologically confirmed lung cancer were formalin-fixed and paraffin-embedded for alignment. First put the tissue chip into the oven, adjust the temperature to 63 degrees, and bake the wax for one hour. Dewaxing with xylene and dehydration with graded alc...

Embodiment 2

[0049] Preparation and characterization of embodiment 2 folic acid liposome and liposome complex

[0050] In this experiment, liposomes (PLP) and folic acid-modified liposomes (F-PLP) were prepared by thin film hydration method, and complexed with BIM-S gene to form folate-modified liposome complex (F-PLP / pBIM) . The physicochemical properties of F-PLP / pBIM were measured, especially particle size and zeta potential.

[0051] 1. Materials

[0052] 1,2-Dioleoyl-3-trimethylammonium-propane (chloride salt) (DOTAP) and cholesterol (Chol) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA) and Shanghai Bio Life Science & Technology, respectively Co., Ltd. (Shanghai, China). The mPEG-succinyl cholesterol conjugate (mPEG-suc-Chol) and the folic acid-PEG-succinyl cholesterol conjugate (F-PEG-suc-Chol) were synthesized and purified (see the experimental preparation section). The BIM-S plasmid is constructed by using pVAX as a vector and operably inserting BIM-S coding...

Embodiment 3

[0068] Example 3 In vitro transfection experiment

[0069] 1. Cell culture

[0070] LL / 2 cells and MH-S cells were obtained from the State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University (Chengdu, Sichuan). Cells were incubated at 37°C and 5% CO with Dulbecco's modified Eagle's medium (DMEM) and RPMI1640 medium supplemented with amikacin and 10% fetal bovine serum without folic acid, respectively. 2 cultured in an incubator.

[0071] 2. Experimental method

[0072] F-PLP / eGFP complexes containing 4 μg of eGFP plasmid were used to transfect LL / 2 cells and MH-S cells for 48 hours in a 6-well plate. Transfected cells were stained with Hoechst33342 and photographed using ArrayScanVTI HCS Reader (Waltham, MA, USA). Transfection efficiency was determined using a FACS Calibur flow cytometer (BD Biosciences, San Jose, CA, USA).

[0073] Real-time quantitative RT- PCR detection of BIM gene expression. Approximately 48 hours after transfect...

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Abstract

The invention relates to a folic acid modified liposome complex and a preparation method and use thereof, and belongs to the field of medicine. The folic acid modified liposome complex is prepared from the following raw materials by weight: 1 part of recombinant expression vector loaded with a BIM-S gene and 5 or more parts of folic acid modified liposome, wherein the folic acid modified liposomeis prepared from the raw materials including (2,3-dioleoyloxy-propyl)-trimethylammonium, cholesterol, polyethylene glycol-succinyl-cholesterol and folic acid-polyethylene glycol-succinyl-cholesterol.The invention also provides a preparation method and use of the liposome complex. The folic acid modified liposome complex not only can kill lung cancer cells, but also can affect the tumor microenvironment by targeting TAM, and provides a new drug choice for clinical treatment of lung cancer.

Description

technical field [0001] The invention relates to a folic acid-modified liposome complex, a preparation method and application thereof, and belongs to the field of medicine. Background technique [0002] Lung cancer is the most commonly diagnosed cancer and is the leading cause of cancer death worldwide. According to global cancer statistics, there are approximately 1.8 million new cases of lung cancer each year. The 5-year survival rate is 6% to 14% in male lung cancer patients and 7% to 18% in female patients. Molecularly targeted therapy for lung cancer has been an important area of ​​research over the past few years. The most widely studied types of targeted therapy include blocking overexpressed genes, increasing the activation of tumor suppressor genes, targeting tumor cell antigens, inducing immunity against tumor cell antigens, and inhibiting or altering signals that control tumor cell growth and survival. Most current targeted cancer therapies focus on killing tumo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K9/127A61K47/22A61P35/00
Inventor 魏霞蔚魏于全铁艳
Owner SICHUAN UNIV
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