Oridonin fluorescent probe and its preparation method and use in target cell positioning

A technology of Rubescensin A and fluorescent probes, which is applied in the direction of luminescent materials, biochemical equipment and methods, and the determination/testing of microorganisms, and can solve the incomplete elucidation of the anti-tumor molecular mechanism and molecular effects of Rubescensin A. The mechanism needs to be explored, the target of cell action and the mode of action are not clear, etc., to achieve the effect of mild conditions and simple process

Inactive Publication Date: 2016-10-05
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the anti-tumor molecular mechanism of oridonin has not been fully elucidated, and the intracell...

Method used

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  • Oridonin fluorescent probe and its preparation method and use in target cell positioning
  • Oridonin fluorescent probe and its preparation method and use in target cell positioning
  • Oridonin fluorescent probe and its preparation method and use in target cell positioning

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Synthesis of Oridonin A Fluorescent Probe

[0037] Compound a (1g, 3.83mmol, 1equiv.) and 1,4-butanediol (3.45g, 38.27mmol, 10equiv.) were dissolved in dichloromethane (20.0mL), stirred at room temperature for 18h, concentrated and added to water, Extract with ethyl acetate (30mL×3), combine the organic layers, wash with water, wash with saturated brine, anhydrous Na 2 SO 4 After drying, column chromatography (MeOH / CH 2 Cl 2 1:100, v / v) to obtain the target compound b (1.06 g, 83%). 1 H NMR (300MHz, CDCl 3 ): δ (ppm) 1.23 (t, J = 6.0Hz, 6H), 1.26 (m, 2H), 1.89 (m, 2H), 2.08 (br, 1H), 3.44 (q, J = 6.0Hz, 4H) , 3.73(t, J=6.0Hz, 2H), 4.35(t, J=6.0Hz, 2H), 6.46(d, J=2.4Hz, 1H), 6.61(dd, J=9.0, 2.4Hz, 1H) , 7.36(d, J=9.0Hz, 1H), 8.44(s, 1H); 13 C NMR (CDCl 3 , 75MHz): δ (ppm) 164.0, 158.0, 152.5, 149.0, 130.6, 109.1, 107.2, 96.2, 64.7, 61.5, 44.6, 29.0, 24.5, 12.0; MS (ESI) m / z: 334.3 [M+H] + .

[0038] Dissolve compound b (1g, 3.0mmol, 1equiv.) and glutaric anhydr...

Embodiment 2

[0041] Antiproliferative Activity of Oridonin A Fluorescent Probe

[0042] The tumor cells in the logarithmic growth phase were digested, counted, and prepared at a concentration of 5×10 4 cells / mL of cell suspension, add 100 μl of cell suspension to each well of a 96-well plate (5×10 per well 3 cells). Place the 96-well plate at 37°C, 5% CO 2 After culturing in the incubator for 24 hours, the drug was diluted to the desired concentration with complete medium, and 100 μL of the corresponding drug-containing medium was added to each well. Place the 96-well plate at 37°C, 5% CO 2 After culturing in the incubator for 72 hours, stain the 96-well plate with MTT; add 20 μL MTT (5 mg / mL) to each well, and continue culturing in the incubator for 4 hours; discard the medium, add 150 μL DMSO to each well to dissolve, and shake gently for 10 minutes. Mix well; λ=490nm, read the OD value of each well with a microplate reader, and calculate the inhibition rate. Experimental results (s...

Embodiment 3

[0043] Example 3 Rubescensin A fluorescent probe staining of HepG2 cells

[0044] HepG2 cells in the logarithmic growth phase were selected and seeded on glass coverslips placed in a 6-well plate, 5×10 per well 5 After 24 hours of attachment, compound d (1 μM in DMSO) was added to the complete medium and incubated at different time points (0, 5, 10, 15, 30, 60 minutes). After incubation for the corresponding time, cells were fixed with 4% paraformaldehyde (in 1×PBS) for 10 minutes, and after washing with PBS, the glass coverslips were removed for fluorescence imaging. Under the fluorescent microscope, observe the coloring part of the cells, the distribution of fluorescence and the change of brightness, etc., the results are shown in figure 2 . It can be seen from the figure that the oridonin fluorescent probe can quickly enter the cells.

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Abstract

The invention relates to a novel oridonin fluorescent derivative and concretely relates to an oridonin fluorescent probe prepared by connecting a fluorophore to an oridonin framework. The invention also discloses a preparation method of the oridonin fluorescent derivative and a potential use of the oridonin fluorescent derivative in antitumor mechanism research and cancer treatment and especially relates to a use of the oridonin fluorescent probe in detection of oridonin target cell positioning.

Description

technical field [0001] The present invention relates to the preparation of anti-tumor activity natural product oridonin A fluorescent probe derivatives, in particular to the application of this type of fluorescent probe in detecting oridonin A in the localization of target cells. Background technique [0002] Prevention and treatment of tumor diseases is the dream of all mankind, and scientists are working hard and unremittingly for this purpose. With the rapid development of life science research, people's research on tumor mechanism is deepening, and many anti-tumor drugs have been developed, of which natural drugs account for a considerable proportion. Although natural products have good anti-tumor effects, the anti-tumor mechanism of many natural products is still unclear, which limits the clinical application of these natural products. Therefore, it is urgent to explore the anti-tumor mechanism of natural products. [0003] my country is rich in enantio-kaurene diterpe...

Claims

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Application Information

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IPC IPC(8): C07D493/08C09K11/06A61K31/352A61K47/48A61P35/00C12Q1/02
Inventor 徐进宜徐盛涛姚鸿罗姗姗王诚倩张晨曦李达翃裴玲玲王光雨胡梅
Owner CHINA PHARM UNIV
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