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Tumor cell membrane drug loading system as well as construction method and application thereof

A construction method and cell membrane technology are applied to the tumor cell membrane drug-carrying system of long-circulating polypeptide drugs in vivo and the field of construction thereof, which can solve the problems of inability to produce effective blood drug concentration, low oral bioavailability, and reduced patient compliance, etc. Enhanced MRI effect, good biocompatibility, and increased concentration

Pending Publication Date: 2022-01-04
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although peptide drugs are made into sustained-release microsphere formulations, their oral bioavailability is still very low, so that they cannot produce high enough effective blood drug concentrations
[0006] However, when subcutaneously injecting polypeptide drugs, due to the presence of proteases in the body, the half-life of the drug in the body is very short, requiring frequent injections, which will increase the pain of the patient and reduce the patient's compliance

Method used

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  • Tumor cell membrane drug loading system as well as construction method and application thereof
  • Tumor cell membrane drug loading system as well as construction method and application thereof
  • Tumor cell membrane drug loading system as well as construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] This embodiment provides a tumor cell membrane drug-carrying system, the preparation method of which includes the following steps:

[0107] 1. Preparation of tumor cell membrane

[0108] H460 cells were cultured in RPMI-1640 complete medium containing 10% fetal bovine serum and 1% double antibodies (penicillin and streptomycin) at 37°C in 5% CO 2 cultured in a cell culture incubator until the cell density reached 80%.

[0109] Wash twice with PBS, digest with trypsin diluted 4 times in PBS containing 2mM EDTA, centrifuge at 300g, wash twice with PBS, collect cells and resuspend in hypotonic buffer containing protease inhibitors, rotate at 80rpm at 4°C 1h, sonicate in an ice bath for 30min, sonicate with an ultrasonic cell disruptor for 3min to disrupt the cells, centrifuge at 20,000g for 20min, discard the precipitate, and centrifuge the supernatant at 100,000g for 1h at 4°C to harvest the cell membrane pellet. Then use PBS solution to resuspend, and sonicate for 5 mi...

Embodiment 2

[0135] This example is used to study the nuclear magnetic properties of the cell membrane drug-loaded system.

[0136] The cell membrane prepared in embodiment 1 is wrapped with Fe 3 o 4 The drug-loading system (mem / Fe 3 o 4 ) by Fe 3 o 4 The concentration is made into a series of aqueous solutions with different concentrations, Fe 3 o 4 Concentrations were 20 μg / mL, 10 μg / mL, 5 μg / mL, 1 μg / mL and 0.2 μg / mL, and the Fe 3 o 4 T2 imaging and relaxation time T2 before and after wrapping with cell membrane, different concentrations of Fe 3 o 4 and mem / Fe 3 o 4 The magnetic resonance analysis images of Fe are shown in Figure 6(a) and Figure 6(b). 3 o 4 , mem / Fe 3 o 4 Still has a good imaging effect;

[0137] The relaxation rate r2 of the sample is shown in Figure 6(c), the Fe wrapped in the cell membrane 3 o 4 There is some decrease in the relaxation rate, which may be due to the coating of the cell membrane covering the Fe to a certain extent 3 o 4 imaging effe...

Embodiment 3

[0139] This example detects and studies the cytotoxicity of the cell membrane drug-loaded system.

[0140] Naked Fe 3 o 4 and membrane-wrapped Fe 3 o 4 Drug loading system, according to Fe 3 o 4 The amount of CCK8 (CellCounting Kit-8) reagent was added to each well after being co-cultured with H460 cells in a 37°C cell culture incubator for 24 hours. The standard instrument detects the absorbance at 450nm.

[0141] The result is as Figure 7 As shown, the exposed Fe 3 o 4 No cytotoxicity, and the Fe after cell membrane wrapping 3 o 4 It has stronger biocompatibility and can better promote cell proliferation.

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Abstract

The invention provides a tumor cell membrane drug loading system as well as a construction method and application thereof. The drug loading system comprises a tumor cell membrane and a polypeptide drug connected to the surface of the tumor cell membrane. The tumor cell membrane has good biocompatibility, so that the half-life period of the polypeptide drug in vivo can be prolonged; the tumor-associated antigen capable of being expressed on the surface of a tumor cell membrane can play a role of a tumor vaccine after being phagocytized by macrophages; homogeneous adhesion antigens can be expressed on the surfaces of tumor cells, so that the drug loading system is actively targeted to tumor parts, and active targeted drug delivery is realized. Meanwhile, the contrast agent is wrapped in the tumor cell membrane, and the contrast agent is gathered at the tumor part and the concentration of the contrast agent is increased by utilizing the active targeting property of the tumor cell membrane to the tumor part, so that the nuclear magnetic imaging effect of the tumor part is enhanced. Therefore, the tumor cell membrane drug loading system has important significance for realizing tumor diagnosis and treatment integration.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a tumor cell membrane drug-carrying system and its construction method and application, in particular to a tumor cell membrane drug-carrying system of long-circulating polypeptide drugs in the body, its construction method and application. Background technique [0002] Cancer has become an increasingly serious health problem, and immunotherapy has become an effective means of treating cancer. Antibody drugs with high specificity, high sensitivity, and good stability have played an important role in immunotherapy, but they have the disadvantages of high cost, large batch-to-batch variance, and high immunogenicity. [0003] With the maturity of biotechnology and peptide synthesis technology, more and more peptide drugs have been developed and applied clinically. Compared with antibody drugs, peptide drugs have wide indications, low cost, small batch-to-batch variation, low im...

Claims

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Application Information

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IPC IPC(8): A61K47/69A61K49/18A61K38/16A61P35/00
CPCA61K47/6903A61K49/1896A61K38/16A61P35/00
Inventor 朱毅敏李霖王姣姣孟祥州郄博
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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