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Photodynamic combined calcium overload nano-carrier and preparation method and application thereof

A nanocarrier and antibody technology, applied in the field of biotechnology and biomedicine, can solve the problems of physical harm, the measurement cannot kill tumors, and the radiation dose is difficult to control, and achieve the effect of promoting apoptosis.

Pending Publication Date: 2021-12-10
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional surgical resection cannot completely remove the lesion, which lays a hidden danger for postoperative recurrence and unsatisfactory prognosis
Clinical treatment mostly uses synergistic means of surgery, chemotherapy, and radiotherapy, but the current chemotherapy method is extremely harmful to the body due to the nature of the drug itself and the drug delivery mechanism in the body, and often brings more serious side effects
Radiation therapy is difficult to control because of its radiation dose, too small dose can not achieve the effect of killing tumors, and too large dose can cause damage to normal tissues, so the general treatment of tumors is still in a dilemma

Method used

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  • Photodynamic combined calcium overload nano-carrier and preparation method and application thereof
  • Photodynamic combined calcium overload nano-carrier and preparation method and application thereof
  • Photodynamic combined calcium overload nano-carrier and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1: BPQD@CaO 2 - Synthesis of PEG-GPC3Ab nanocarriers.

[0062] This example is mainly for the synthesis of BPQD@CaO with good shape and suitable size 2 - PEG-GPC3Ab nanocarriers. details as follows:

[0063] 1. Preparation of black phosphorus quantum dots and calcium peroxide

[0064] ①Sonicate 10 mL of black phosphorus nanosheet (BPNs) mixture (200 μg / mL) dispersed in aqueous solution for 12 h (or may be extended appropriately), and keep the temperature below 5 °C in an ice-water bath;

[0065] ② The frequency of ultrasonic treatment is 20kHz, the ultrasonic head works for 5s, and the interval between each cycle is 5s. ;

[0066] ③Then the sonicated mixture was centrifuged at 13000rpm for 30min. ;

[0067] ④ The primary product of black phosphorus quantum dots (BPQDs) was washed three times with ultrapure water and stored at 4 °C for future use;

[0068] ⑤ Add 1.0g calcium hydroxide (Ca(OH) 2 ) was added to 20mL ultrapure water, and then ultrasonically...

Embodiment 2

[0076] Physicochemical property characterization of embodiment 2 nano drug delivery carrier

[0077] BPQDs and CaO by FEI Tecnai G2 transmission electron microscope 2 The morphology of the nano-based nanoparticles was characterized; sample preparation method: the sample was dispersed to a reasonable concentration with water, and then a small drop of the solution was dropped onto a copper grid to dry overnight. see results figure 1 .

Embodiment 3B

[0078] Example 3BPQD@CaO 2 - PEG-GPC3Ab targeting liver cancer cell Huh7 experiment

[0079] In cell targeting experiments, Fluo-4 AM is used to detect fluorescence intensity to indirectly represent calcium content.

[0080] ① Seed cells of appropriate density on glass slides and incubate at 37°C, 5% CO 2 Under culture for 24h;

[0081] ② Then, based on CaO 2 The nanoparticles were added to the medium, and then incubated for 2h;

[0082] ③Then according to the standard experimental procedure: wash with PBS three times after removing the medium;

[0083] ④ Then fix with PFA for 15min, then wash away PFA with PBS, stain the cells with DAPI for 5-10min, and measure the fluorescence by confocal laser scanning microscope (CLSM), see figure 2 . The experimental results show that the BPQD@CaO prepared by the present invention 2 - PEG-GPC3Ab nanocarriers can target tumor cells.

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Abstract

The invention relates to a photodynamic combined calcium overload nano-carrier and a preparation method and application thereof, and particularly discloses a tumor cell targeted anti-tumor nano-carrier. The tumor cell targeted anti-tumor nano-carrier is characterized in that the nano-carrier comprises calcium peroxide particles, black phosphorus quantum dots are electrostatically adsorbed on the surfaces of the calcium peroxide particles. The calcium peroxide particles adsorbed with the black phosphorus quantum dots are wrapped by a polymer to form nanoparticles, and the polymer is coupled with a modifier of a targeted tumor surface marker; the modifier is selected from an antibody, folic acid, polypeptide and an aptamer of the targeted tumor surface marker. According to the photodynamic combined calcium overload nano-carrier and the preparation method and application thereof, the nano-carrier can promote apoptosis of tumor cells with high expression of GPC3 or inhibit proliferation of the tumor cells, can be used as a biological nano-drug of a GPC3 binding site, and can be used for preparing a drug for preventing and / or treating liver cancer.

Description

technical field [0001] The invention relates to the fields of biotechnology and biomedicine, in particular to a photodynamically combined calcium-overloaded nanocarrier and a preparation method and application thereof. Background technique [0002] 1.1. Liver cancer [0003] Hepatocellular carcinoma (HCC) is a common malignant tumor worldwide, and more than 53% of HCC patients are from China. Currently, the main curative treatment for HCC is surgical resection, which is also the first choice for patients with early-stage HCC whose tumor size is less than 3 cm. However, current treatment outcomes are still unsatisfactory, partly due to postoperative tumor recurrence. It has been reported that approximately 50% of patients may experience tumor recurrence, manifested as intrahepatic or extrahepatic metastases within 2 years after curative hepatectomy. Therefore, exploring the molecular mechanism of HCC recurrence is still of great significance for improving clinical prognosi...

Claims

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Application Information

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IPC IPC(8): A61K47/69A61K41/00A61K45/00A61K47/54A61K47/62A61K47/68B82Y5/00B82Y30/00B82Y40/00A61K31/341A61K33/40
CPCA61K41/0057A61K31/341A61K33/40A61K47/6935A61K47/6937A61K47/6939A61K47/62A61K47/6835A61K47/545A61K47/549A61K45/00B82Y5/00B82Y30/00B82Y40/00A61K2300/00
Inventor 黄来强过冬冬代小勇王坤王丽君冯春燕刘昱宏
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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