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Use of brusatol and analogues thereof in treatment of pituitary adenoma

A technology of Brucea Brucea and pituitary adenoma, which is applied in the field of medicine and can solve the problems that pituitary adenoma has not been reported.

Active Publication Date: 2021-04-13
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] However, there is no report on the application of Brucea Brucea in the treatment of pituitary adenoma at home and abroad at present.

Method used

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  • Use of brusatol and analogues thereof in treatment of pituitary adenoma
  • Use of brusatol and analogues thereof in treatment of pituitary adenoma
  • Use of brusatol and analogues thereof in treatment of pituitary adenoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Brusatol inhibits cell activity and hormone secretion of pituitary adenomas

[0041] 1) Take the GH3 cells and MMQ cells in the logarithmic growth phase in good condition, wash 2-3 times with PBS, and use F12 medium at 37 ° C, 5% CO 2 cultured under the conditions of 2.5% fetal bovine serum, 15% horse serum and 100U / ml penicillin / streptomycin; take fresh tumor specimens from neurosurgical pituitary adenoma surgery in the First Affiliated Hospital of Wenzhou Medical University, in Under sterile conditions, the tumor tissue was cut into pieces with ophthalmic scissors, and then the tumor tissue was mechanically digested with type IV collagenase and trypsin to obtain primary pituitary tumor cells. The culture method was the same as that of GH3 and MMQ cells.

[0042] 2) Inoculate in the culture plate at a ratio of 100,000 to 200,000 cells / ml medium, and culture in a 96-well plate. The medium in each well is 100 μl for the detection of cell activity experiments; ...

Embodiment 2

[0048] Example 2 Verification test of Brusatol on subcutaneous tumor formation of pituitary adenoma in nude mice

[0049] 1) Purchase 5-week-old female BALB / c(nu / nu) nude mice from Shanghai Experimental Animal Center, and feed them adaptively in an SPF animal room for one week. When the nude mice have no obvious abnormalities or discomfort, GH3 cells and MMQ can be performed. Tumor formation experiment of subcutaneous implantation of cells.

[0050] 2) The inoculation site is subcutaneous in the lower back, avoiding the forelimbs, not touching the muscles, and inoculating 2×10 6 GH3 and MMQ cells were inoculated on one side of a nude mouse. Use F12 medium, add 2.5% fetal bovine serum, 15% horse serum and 100U / ml penicillin / streptomycin to resuspend the cells, and wipe and disinfect the needle insertion site with 75% alcohol before inoculation.

[0051] 3) Take out the cell suspension with a disposable needle, remove the air, puncture about 1 cm forward from the needle insert...

Embodiment 3

[0053] Example 3 Verification of Brusatol-induced apoptosis in pituitary adenoma cells

[0054] 1) Take the GH3 cells and MMQ cells in the logarithmic growth phase in good condition, wash 2-3 times with PBS, and use F12 medium at 37 ° C, 5% CO 2 cultured under the conditions of 2.5% fetal bovine serum, 15% horse serum and 100U / ml penicillin / streptomycin.

[0055] 2) Inoculate in the culture plate at a ratio of 300,000 cells / ml culture medium, use a 6-well plate for culture, and the medium in each well is 2ml; then add Brusatol in the 6-well plate according to the working concentration of 0.5 μM as above, and culture After 24 hours, the cells were collected in 1.5 mL EP tubes and placed on ice.

[0056] 3) The proportion of apoptotic cells in GH3 and MMQ cells treated with Brusatol was detected by flow cytometry with Annexin-V and FITC double staining. The results showed that the proportion of apoptotic cells after Brusatol treatment increased significantly.

[0057] 4) Take ...

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Abstract

The invention discloses use of brusatol and analogues thereof or pharmaceutically acceptable salts thereof in preparing medicines for treating pituitary adenoma. Different from a classic approach of promoting tumor cell apoptosis by DRD2 dependence of a dopamine receptor agonist, the brusatol and analogues thereof achieve inhibition effects of medicine on pituitary adenoma such as prolactinoma, clinical non-functional pituitary adenoma and growth hormone adenoma by promoting cells to generate ROS to induce cell apoptosis.

Description

technical field [0001] The invention relates to the field of medicines, in particular to a new application of Brucea javanica extract, Brucea brucea picrol or analogues thereof, in the treatment of human pituitary adenomas. Background technique [0002] Pituitary adenoma is the third most common intracranial tumor, with an incidence rate of 7.5-15 per 100,000 people. In recent years, the incidence of pituitary adenoma has been increasing. A recent Belgian epidemiological survey found that the incidence of pituitary adenoma was as high as 75-100,000. 113 / 100,000, and the autopsy discovery rate is even as high as 20%-30%. Pituitary adenomas are mainly divided into non-functioning adenomas, prolactinomas, growth hormone adenomas, ACTH adenomas, etc., and prolactinomas are the most common pituitary adenomas, accounting for 40% to 45% of all pituitary adenomas . The clinical manifestations of patients with prolactinoma are mainly amenorrhea and galactorrhea, sexual dysfunction,...

Claims

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Application Information

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IPC IPC(8): A61K31/366A61P35/00
CPCA61K31/366A61P35/00
Inventor 苏志鹏吴泽睿蔡霖卢江龙李群王成德
Owner THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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