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Preparation method of propranolol stealth liposome modified by vascular endothelial growth factor receptor-2 (VEGFR-2) monoclonal antibody and casein phosphopeptides (CPP) jointly, and products thereof

A stealth lipid technology of VEGFR-2 and naphthalene, applied in liposome delivery, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the abnormal development of tissues and organs and the lack of targeting Therapeutic effect, increased patient pain and other issues

Inactive Publication Date: 2013-08-14
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of targeted therapeutic effect of urea in the treatment of hemangioma, it is difficult to form high-concentration accumulation in the tumor body, and multiple injections are required, which prolongs the treatment cycle and increases the suffering of patients; and because the concentration of urea is too high, the dosage per unit time is too large , local injection can cause skin tissue necrosis, and can also cause developmental deformities of adjacent normal tissues and organs

Method used

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  • Preparation method of propranolol stealth liposome modified by vascular endothelial growth factor receptor-2 (VEGFR-2) monoclonal antibody and casein phosphopeptides (CPP) jointly, and products thereof
  • Preparation method of propranolol stealth liposome modified by vascular endothelial growth factor receptor-2 (VEGFR-2) monoclonal antibody and casein phosphopeptides (CPP) jointly, and products thereof
  • Preparation method of propranolol stealth liposome modified by vascular endothelial growth factor receptor-2 (VEGFR-2) monoclonal antibody and casein phosphopeptides (CPP) jointly, and products thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Establish the HPLC-FID analysis method of measuring body fluid propranolol concentration:

[0040] The concentration of propranolol in blood, tissue and cell culture medium was determined by HPLC-FID (fluorescence) detection method.

[0041] Specific method: Carvedilol was used as internal standard. Detection wavelength: excitation light 290nm, emission light 340nm. Chromatographic column: C18 column (250×4.6mm, 2.5μm). Add 10 microliters of internal standard solution to biological samples, add 50 microliters of saturated sodium carbonate and 2.5mL of a mixed solvent (n-hexane-dichloromethane-isopropanol=65:30:5) and vortex for 3 minutes, centrifuge at 4000rpm for 10 minutes, and separate The supernatant was blown dry with nitrogen, reconstituted in 100 μL methanol and injected 50 μL for determination.

Embodiment 2

[0043] Determination of the composition and ratio of complex phospholipids: using distearoylphosphatidylcholine (DSPC) and hydrogenated soybean phosphatidylcholine (HSPC) as high phase transition temperature phospholipids; using dipalmitoylphosphatidylcholine (DPPC) and dimyristoyl Phosphatidylcholine (DMPC) is a phospholipid with a low phase transition temperature, and two combinations are used to prepare complex phospholipid liposomes (a total of 4 combinations). In the case of adding the stealth modification material DSPE-PEG2000, different combinations and different combinations under the same combination are compared. The effect of phospholipid ratio on drug loading of propranolol and its stability in plasma (that is, the leakage rate of propranolol). Finally, the composition and ratio of the complex phospholipids with good drug-loading effect and stability are determined for the following liposome research. The optimal ratio is: distearoylphosphatidylcholine (DSPC), hydr...

Embodiment 3

[0045] Preparation of complex phospholipid liposomes: prepared by thin film-ultrasonic method. Weigh 120mg of phospholipid material and dissolve in 10mL of chloroform, put it in a 50mL eggplant-shaped bottle and evaporate it under reduced pressure at 60°C to form a thin film, take out the eggplant-shaped bottle, and place it in a vacuum drying oven at 55°C for 12h. Add 5mL of ammonium sulfate to the eggplant-shaped bottle, and rotate and hydrate at 60°C for 40min. The obtained liposome suspension is ultrasonicated (400W, 50 times) in a water bath at 60°C, centrifuged at 1000rpm at low speed to remove titanium powder, and dialyzed with pH7.4PBS to remove the outer layer. Water phase ammonium sulfate, add propranolol powder and mix, 60 ℃ 20min, that is. Sephadex G-50 column (inner diameter 1cm, length 27cm) on drug-loaded liposomes, eluted with pH 7.4 PBS, flow rate 1mL min -1 , the liposome part and the free part were collected after loading on the column, and the concentratio...

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Abstract

The invention discloses a preparation method of a propranolol stealth liposome modified by a vascular endothelial growth factor receptor-2 (VEGFR-2) monoclonal antibody and casein phosphopeptides (CPP) jointly; the method comprises the following steps of: (1) confirming the composition and the proportion of liposome compound phosphatide; (2) preparing a propranolol stealth liposome modified by the CPP; and (3) connecting the VEGFR-2 monoclonal antibody and the surface of the propranolol stealth liposome modified by the CPP to obtain the propranolol stealth liposome modified by the VEGFR-2 monoclonal antibody and the CPP jointly. The preparation method has the characteristics of high stability, long circulation, active targeting, high-efficiency endocytosis and the like, and can guarantee the toxicity reducing and efficacy enhancing of anti-hemangioma medicines. The research result of the method provides a preparation technology platform for the clinical targeted therapy of the anti-hemangioma medicines.

Description

technical field [0001] The invention relates to the fields of molecular cloning and biomedicine, in particular to a preparation method of propranolol stealth liposomes modified in combination with VEGFR-2 monoclonal antibody and CPP, as well as its products and applications. Background technique [0002] Hemangiomas are common benign tumors, and there are obvious racial and gender differences in their incidence, and their incidence may be higher in low birth weight and premature infants. The potential physical and psychological harm of hemangiomas to patients, especially children, cannot be ignored. Especially for hemangiomas in special locations and types, active intervention and treatment must be taken during the proliferation period. Scholars have proposed that the main mechanism of hemangioma is caused by the imbalance of pro-angiogenic factors and inhibitory factors. Angiogenesis is an important factor in the formation of hemangiomas, the main feature of which is the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P35/00A61K31/138A61K47/48A61K47/24A61K47/42A61K9/127
Inventor 李俊
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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