Method for preparing sPD1-loaded microbubbles targeting CD133 and application thereof in preparing drugs for inhibiting cervical cancer

A microbubble, targeted technology, applied in the field of lipid microbubbles, can solve the problems of obvious systemic side effects, easy generation of drug resistance, and difficult treatment.

Active Publication Date: 2019-10-18
CHINA THREE GORGES UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgery is one of the main methods for the treatment of cervical cancer. Although surgery can temporarily remove the lesion, the tumor cells are highly invasive and metastatic, and the lesion cannot be completely removed.
Adjuvant chemotherapy is widely used in preoperative and postoperative treatment in clinical practice. It can effectively reduce tumor staging, reduce parametrial invasion, and provide good conditions for surgery. However, chemotherapy is poorly targeted, prone to drug resistance, and systemic toxicity. Obvious side effects
Cervical cancer is a radiosensitive tumor. The effect of radiotherapy on advanced patients and recurrent patients is affirmed, but its serious complications bring great difficulties to the treatment

Method used

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  • Method for preparing sPD1-loaded microbubbles targeting CD133 and application thereof in preparing drugs for inhibiting cervical cancer
  • Method for preparing sPD1-loaded microbubbles targeting CD133 and application thereof in preparing drugs for inhibiting cervical cancer
  • Method for preparing sPD1-loaded microbubbles targeting CD133 and application thereof in preparing drugs for inhibiting cervical cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Preparation and Characterization of Microbubbles Carrying sPD1 Targeting CD133

[0035]1) Distearoylphosphatidylcholine (DSPC), distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), distearoylphosphatidylethanolamine-polyethylene glycol 2000-biotin (DSPE-PEG2000-Biotin), stearic acid-polyethylimide (PEI-600) were mixed in a glass test tube at a molar ratio of 85:5:5:5, shaken gently, mixed well, and preheated in a 60°C water bath After heating, place the glass test tube on a vortex shaker and rinse with N 2 Blow the liquid in the glass test tube and vortex while blowing to form a homogeneous film at the bottom of the glass tube wall.

[0036] 2) Connect the glass tube after film formation to a vacuum pump, and vacuum for 3 hours to fully volatilize the liquid.

[0037] 3) After taking out the glass test tube, add 5ml of Tris buffer solution to it, put it in an ultrasonic cleaner for 5 minutes in a water bath, so that the film on the glass tube wa...

Embodiment 2

[0053] Application of microbubbles carrying sPD1 targeting CD133 in the preparation of drugs for inhibiting cervical cancer

[0054] Establishment of subcutaneous xenograft tumor model of U14 cervical cancer in mice

[0055] Ten days after the U14 cells were inoculated in the right anterior armpit of the mice, tumors were formed, most of which were round, with a diameter of about 7 cm, and a few were irregular. 50 mice were used in the experiment, and 45 mice were successfully modeled, with a tumor formation rate of 90%. The mice were growing well, and no abnormal death was found. The model mice were randomly divided into five groups, with 9 mice in each group.

[0056] Inhibitory effect of ultrasound combined with targeted gene-loaded microbubbles on cervical cancer in mice

[0057] Tumor inhibition rate of mice in each group

[0058] The mouse developed a bulging mass in the right armpit. Tumor tissues excised from the axilla of mice were nodular, slightly firm in textu...

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Abstract

The invention relates to a method for preparing ultrasound-assisted sPD1-loaded microbubbles targeting CD133. The method specifically comprises the steps that PE labeled streptavidin is added dropwiseinto biotinylated lipid microbubbles for incubation, then a biotinylated CD133 antibody is added, and after the incubation is completed, centrifugation is carried out to obtain the microbubbles targeting CD133; the obtained microbubbles targeting CD133 and sPD1 plasmids are incubated at room temperature, then washed with PBS and centrifuged to obtain the sPD1-loaded microbubbles targeting CD133.The sPD1-loaded lipid microbubbles targeting CD133 are used for preparing drugs for inhibiting cervical cancer transplantation tumors, results show that the sPD1-loaded lipid microbubbles targeting CD133 have a higher antitumor effect, and the volume inhibition rate and body weight inhibition rate of the sPD1-loaded lipid microbubbles targeting CD133 are 78.01% and 72.25% respectively.

Description

technical field [0001] The invention relates to a lipid microvesicle, specifically, ultrasound combined with CD133-carrying sPD1 microvesicle, which is applied to the preparation and treatment of cervical cancer. Background technique [0002] Cervical cancer is one of the most common gynecological malignancies in women all over the world. Its incidence rate is second only to breast cancer and ranks second in gynecological malignancies. Cervical cancer is mainly caused by human papillomavirus (HPV) infection. At present, it is believed that the interaction between persistent high-risk HPV infection, decreased immune function of the body and the local microenvironment of cervical cancer promotes the occurrence and development of cervical cancer. Current treatment options include surgical resection, radiotherapy, and chemotherapy. Surgery is one of the main methods for the treatment of cervical cancer. Although surgery can temporarily remove the lesion, the tumor cells are hig...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K9/127A61K47/42A61K47/34A61P35/00A61K49/22
CPCA61K48/005A61K9/1273A61K47/42A61K47/34A61P35/00A61K49/223Y02P20/55
Inventor 赵云刘朝奇郑智唯
Owner CHINA THREE GORGES UNIV
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