CN104327152A - Triptolide derivatives and application thereof - Google Patents
Triptolide derivatives and application thereof Download PDFInfo
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- CN104327152A CN104327152A CN201410538497.2A CN201410538497A CN104327152A CN 104327152 A CN104327152 A CN 104327152A CN 201410538497 A CN201410538497 A CN 201410538497A CN 104327152 A CN104327152 A CN 104327152A
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- triptolide
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J73/00—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms
- C07J73/001—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom
- C07J73/003—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom by oxygen as hetero atom
Abstract
Triptolide derivatives and application thereof relate to triptolide. Triptolide and the derivatives thereof has different-degree anticancer activity on HepG2 cell and MCF-7 cell. A method for using tRXRalpha and RXRalpha to evaluate the anticancer activity and toxic and side effects of triptolide and the derivatives thereof is firstly provided. The triptolide and the derivatives thereof are confirmed to have the anticancer activity closely related to the tRXRalpha protein expression lowering capacity, and the stronger rRXRalpha lowering capacity, the higher anticancer acitivty. Also the technical scheme firstly confirms that reduction of the toxic and side effects of the triptolide derivatives is related to the stability of normal-cell full-length RXRalpha protein, and the stronger protection effect of the derivatives on full-length RXRalpha, the smaller toxic and side effects.
Description
Technical field
The present invention relates to triptolide, especially relate to triptolide derivative and application thereof.
Background technology
Retinol X receptor (Retinoid X Receptor, RXR) on-steroidal hormone receptor superfamily is belonged to, the unique member of in nuclear receptor family one, can not only homodimerization, and Heterodimerization can be formed with other nuclear receptors (as RAR, PPAR, VDR, TR and Nur77 etc.).Therefore, RXR α plays very crucial effect, the expression of RXR α and the change of function in the biological function regulating Growth of Cells, differentiation and apoptosis, participates in the generating process of numerous disease and cancer.RXR alpha expression amount reduces relevant with the formation of malignant tumour, as t cell lymphoma, thyroid carcinoma, prostate cancer and nonsmall-cell lung cancer.In adult tissue, RXR α melts and causes skin and prostatic precancerous lesion.Moreover, the phosphorylation by changing RXR α is relevant with the generation of liver cancer and colorectal carcinoma.RXR α is attached to PML/RAR, and to be that acute promyelocytic leukemic is formed necessary, proves that abnormal RXR α has oncogenic potential further.In a lot of cancer cells, by proteolytic enzyme cutting RXR α, and then change its function.The applicant reported that the tRXR α albumen having N-end to shorten in many cancer cells and infantile tumour occurred recently, but cancer is other and do not exist in healthy tissues.RXR α unlike total length is mainly positioned at nucleus, and tRXR α is positioned at tenuigenin, interacts with the subunit p85 α of PI3K, and this promotes the signal path of tumor growth, generation and resistance to activate PI3K/AKT.Therefore, the rise of tRXR α in cancer cells, and the New function different from RXR α, imply that can be innovation target spot based on tRXR α, screen and develop new antitumor drug.
Triptolide (Triptolide) is the crystalline monomer compound obtained by Celastraceae plant tripterygium root (peeling) extraction, purifying, it is a kind of diterpene with abietane skeleton, and containing unique three epoxy constructions and α, a β-unsaturated lactone ring five membered.Triptolide is one of trypterygine main active ingredient, there is antitumor, anti-inflammatory, the multiple pharmacological effect such as immunosuppression and antifertility, be widely used in treatment autoimmune disorder, organ transplantation etc., but the target spot of its cell death inducing and mechanism are still unclear.
Research display, triptolide has the effect activating p53 apoptosis path, thus the cracking of induction Bcl-2 albumen, make the apoptosis of mitochondrial.Meanwhile, triptolide can also reduce short growth factor and lifetime because of expression, as cyclin D1 and bcl-X.In addition, triptolide can also reduce the expression of heat shock protein(HSP), as Hsp70, and the inactivation (HSF) of mediation Features of The Heat Shock Transcription Factor, and can the generation of inflammation-inhibiting.But triptolide has its weak point as cancer therapy drug: its poorly water-soluble, and target spot specificity is not high, and toxicity is larger.Investigator both domestic and external carries out structural modification a large amount of work to triptolide, wishes to synthesize target spot specificity high, the tumor that good effect, toxicity are low.
Summary of the invention
The first object of the present invention is to provide the triptolide derivative that a kind of toxicity reduces, antitumour activity is high.
The second object of the present invention is to provide a kind of triptolide derivative and is preparing the application in cancer therapy drug.
The third object of the present invention is to provide the application of a kind of triptolide derivative in antitumour activity and evaluating toxic and side effect.
Described triptolide derivative, its structural formula is:
Wherein, R takes from amine, various lipid acid or aromatic amine, and R can be the one in tetramethyleneimine (1), methyl piperidine (2), butylamine (3), thanomin (4), chloroaniline (5), piperazine (6), diethanolamine (7) etc.; R-structural formula is as shown in table 1.
Table 1
Research finds, triptolide can target tRXR α-PI3K/AKT signal path inducing apoptosis of tumour cell and Tumor suppression growth, and this acts under the help of TNF α and is exaggerated, and plays more special antitumous effect.
Described triptolide derivative is through cell MTT experiment, Western blot experiment and immunofluorescence technique etc., show triptolide derivative of the present invention, i.e. compound shown in logical formula I, all there is anti-tumor activity in various degree, comprise hepatoma cell line (HepG2), breast cancer cell line (MCF-7).Detect through reporter gene and Western blot, triptolide derivative antitumour activity of the present invention is relevant with selective degradation tRXR α, and the reduction of toxic side effect is relevant with stablizing of total length RXR α.As can be seen here, triptolide derivative can be applied preparing in cancer therapy drug, and described cancer therapy drug includes but not limited to anti-hepatoma cell line (HepG2) medicine, breast cancer cell line (MCF-7) medicine etc.
In addition, described triptolide derivative can be applied in antitumour activity and evaluating toxic and side effect.
The present invention relates to and assess the antitumous effect of triptolide derivative and the application of toxic side effect based on RXR α/tRXR α target spot.Described triptolide and derivative thereof have antitumour activity in various degree to HepG2 cell and MCF-7 cell.Provide the method that a kind of tRXR α and RXR α assesses triptolide and derivative antitumour activity and toxic side effect first.The present invention confirms that the ability of triptolide and derivative antitumour activity and downward tRXR α protein expression is closely related, and the ability lowering tRXR α is stronger, and antitumour activity is stronger.The present invention also confirms that the reduction of triptolide derivative toxic side effect is relevant with its stabilizing power for normal cell total length RXR α albumen first, and derivative is stronger for the provide protection of total length RXR α, and its toxic side effect is less.
Compared with prior art, feature of the present invention is that to report and study more natural compounds triptolide be lead compound, its C-14 position is carried out to modification and the transformation of nitrogen-containing group, thus obtains a class and have anticancer compound in various degree.Meanwhile, present invention also offers a kind of with the antitumous effect of the tRXR α assessment triptolide that is target spot and derivative thereof, evaluate the application of its toxic side effect with RXR α.
Accompanying drawing explanation
Fig. 1 represents the dose curve of triptolide (TR01) and derivative 1 ~ 4 T suppression cell vigor thereof.
Fig. 2 represents the schematic diagram that triptolide (TR01) and derivative 1 ~ 7 thereof affect HepG2 cell PARP cutting, tRXR α and RXR alpha levels.
Embodiment
Below in conjunction with specific embodiment, the present invention is described further, but specific embodiment does not do any restriction to the present invention.
Embodiment 1: the dose curve of triptolide and derivative T suppression cell vigor thereof and IC
50pH-value determination pH
The present embodiment adopts mtt assay to measure triptolide derivative to the impact of various cancer cell multiplication, and its concrete operations are as follows:
(1) experiment material
Clone: HepG2 (ATCC HB-8065), MCF-7 (ATCC HTB-22).
Cancerous cell line DMEM (Gibco company)+10% foetal calf serum (Hyclone company) is cultivated.Experiment also needs triptolide and derivative (oneself synthesis) thereof, 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide bromine salt (Sigma).
(2) experimental technique
Have restraining effect with mtt assay detection triptolide and the propagation of derivative to cell thereof, concrete measuring method is as follows:
A. trysinization logarithmic phase cell, collected by centrifugation after stopping, make cell suspension, cell counting adjusts its concentration to 5 ~ 10 × 10
4individual/ml.
B. after being prepared by cell suspension, mix gently, every hole adds 100 μ l, and the density of cell to be measured is like this 5000 ~ 10000/ holes (the aseptic PBS of marginal pore fills).
C. the Tissue Culture Plate inoculated is put into incubator to cultivate, after spending the night, add the medicine of different concns gradient, every hole 100 μ l, if 3 ~ 5 multiple holes.
D.5%CO
2, hatch 16 ~ 48h for 37 DEG C, under inverted microscope, observe the action effect of medicine.
E. every hole adds 10 μ l MTT solution (5mg/ml, i.e. 0.5%MTT), continues to cultivate 4h.
After F.MTT adds cultivation 4h, crystallization can fully be formed.Removed by supernatant, this process will be noted Formazan crystallization can not being removed.Every hole adds 150 μ l dimethyl sulfoxide (DMSO), puts low-speed oscillation 10min on shaking table, crystallisate is fully dissolved.The light absorption value (A value) in each hole is measured at enzyme-linked immunosorbent assay instrument OD490nm place.
G., zeroing hole (substratum, MTT, dimethyl sulfoxide (DMSO)) is set simultaneously, control wells (the medicine dissolution medium of cell, same concentrations, nutrient solution, MTT, dimethyl sulfoxide (DMSO)).
By following formulae discovery cell viability (Cell Viability):
Cell viability=(drug treating group A value-blank group A value)/(control group A value-blank group A value) × 100%.Finally draw cell viability and drug level graph of relation.
H. Graphpad prism 5 is utilized to calculate the IC of each compound
50value.
(3) experimental result:
As can be seen from Fig. 1 and Fig. 2, triptolide derivative still remains the very strong antitumour activity of triptolide, antitumour activity is descending is respectively No. >7, derivative No. 4 >3 >2 >1 >5 >6, the IC of especially No. 4, No. 3 and No. 2
50within remaining on 100nM.
Embodiment 2:Western blot detects triptolide and derivative induced PARP cuts, RXR α and tRXR α degrades
Select hepatocellular carcinoma H22, be incubated in 6 hole tissue culturing plates at 37 DEG C and 5% CO2gas incubator, with the triptolide of 50nM (TR01) and derivative (No. 1, No. 2, No. 3, No. 4, No. 5, No. 6 and No. 7) thereof, serum-free DMEM processes 24h respectively.Then, with RIPA lysis buffer (50mM Tris-HCl, the pH 7.4 improved; 150mM NaCl; 5mM EDTA; 1%NP-40,0.5% Sodium desoxycholate, 0.1%SDS) in cracking 30min on ice.With identical applied sample amount, 8%SDS-PAGE electrophoresis, transferring film, with the TBST of 5% skim-milk (50mM Tris-HCl (pH 7.4), the NaCl and 0.1%Tween-20 of 150mM) room temperature closes 1h, in 4 DEG C of night incubation primary antibodies, the anti-1h of incubated at room two, ECL develops the color, exposure.Finally, PARP, RXR α, tRXR α and β-actin is detected.
As shown in Figure 2, the antitumour activity of compound is closely related with the ability of degraded tRXR α for experimental result.Existing research shows, tRXR α high expression level in cancerous tissue, very large promoter action is played in the formation for tumour.Triptolide (TR01) and partial derivatives (No. 2, No. 3 and No. 4) can be degraded tRXR α in various degree, and cut along with the PARP of respective degrees.But the other a part of derivative of triptolide (No. 5, No. 6 and No. 7) is not only degraded tRXR α, raise tRXR α on the contrary, corresponding short apoptosis function obviously weakens.This means the change based on tRXR α target spot, can be used to the antitumour activity assessing triptolide derivative.
RXR α is the unique member that in nuclear receptor family, has a various biological function, take part in body from fetal development to orga-nogenesis, and numerous process such as the physiological regulating control of the various new katabolism of adult.Visible the existence of cell will directly be had influence on to the disorder of RXR α function in cell.Fig. 2 shows, triptolide (TR01) not only significantly reduces the expression of tRXR α, and significantly reduce the expression of total length RXR α, and No. 2, derivative, No. 3 and No. 4 without obvious degradation effect, even play stabilization to RXR α to total length RXR α.Because total length RXR α is the oligogene maintaining normal cell function, No. 2, derivative, No. 3 may be relevant with its protection for total length RXR α with the reduction of No. 4 toxic side effect.
Triptolide (TR01) and derivative (No. 1, No. 2, No. 3, No. 4, No. 5, No. 6 and No. 7) thereof are to HepG2 cell and MCF-7 cell medium lethal dose IC
50be worth as shown in table 2.
Table 2
The structural formula of triptolide TR01, derivative 1 ~ 8 is as follows:
The antitumour activity of described medicine using tRXR α as action target spot, but does not affect the total length RXR α in normal cell.
Described compound (No. 2, No. 3 and No. 4) inducing apoptosis of tumour cell is by optionally inducing the degraded of tRXR α and suppress AKT to grow path, starting the apoptosis pathway of tumour cell, cause apoptosis of tumor cells.No. 2, No. 3 and No. 4 compounds have significant protective effect to normal cell total length RXR α, and its poison is secondary obviously to be lowered.
Claims (4)
1. triptolide derivative, is characterized in that its structural formula is:
Wherein, R takes from amine, various lipid acid or aromatic amine, and R is the one in tetramethyleneimine (1), methyl piperidine (2), butylamine (3), thanomin (4), chloroaniline (5), piperazine (6), diethanolamine (7); R-structural formula is as follows:
2. triptolide derivative is applied preparing in cancer therapy drug as claimed in claim 1.
3. apply as claimed in claim 2, it is characterized in that described cancer therapy drug includes but not limited to anti-hepatoma cell line (HepG2) medicine, breast cancer cell line (MCF-7) medicine.
4. triptolide derivative is applied in antitumour activity and evaluating toxic and side effect as claimed in claim 1.
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| CN201410538497.2A CN104327152A (en) | 2014-10-14 | 2014-10-14 | Triptolide derivatives and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016181312A1 (en) * | 2015-05-11 | 2016-11-17 | Versitech Limited | Polycyclic epoxides and compositions thereof with anti-cancer activities |
| CN106749496A (en) * | 2016-12-05 | 2017-05-31 | 张奇军 | New triptolide derivative and its preparation and use |
| CN109021061A (en) * | 2018-09-29 | 2018-12-18 | 郭可点 | Triptolide targeted prodrug and its preparation method and application |
| CN110511202A (en) * | 2019-08-13 | 2019-11-29 | 厦门大学 | Polycyclic polyisocyanate pentenyl acyl phloroglucinol class compound and the preparation method and application thereof |
| CN111821308A (en) * | 2019-04-15 | 2020-10-27 | 中国科学院上海药物研究所 | Application of tripterygium wilfordii in preparation of medicine for treating non-alcoholic fatty liver disease |
| CN114478684A (en) * | 2022-01-21 | 2022-05-13 | 南京中医药大学 | Triptolide prodrug, preparation method and medical application thereof |
| WO2023143330A1 (en) * | 2022-01-25 | 2023-08-03 | 杭州卫本医药科技有限公司 | Tripterygium wilfordii diterpene epoxide having function of killing myc positive cell, preparation method therefor, and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101759763A (en) * | 2009-12-29 | 2010-06-30 | 中山大学 | Triptolide derivative, and preparation method and application thereof |
| CN102634561A (en) * | 2012-03-14 | 2012-08-15 | 厦门大学 | Application of triptolide, triptolide derivant and triptolide analogue in preparation of antitumor drugs |
-
2014
- 2014-10-14 CN CN201410538497.2A patent/CN104327152A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101759763A (en) * | 2009-12-29 | 2010-06-30 | 中山大学 | Triptolide derivative, and preparation method and application thereof |
| CN102634561A (en) * | 2012-03-14 | 2012-08-15 | 厦门大学 | Application of triptolide, triptolide derivant and triptolide analogue in preparation of antitumor drugs |
Non-Patent Citations (1)
| Title |
|---|
| FANG XU ET AL: "Design, synthesis, and biological evaluation of novel water-soluble triptolide derivatives: Antineoplastic activity against imatinib-resistant CML cells bearing T315I mutant Bcr-Abl", 《BIOORGANIC & MEDICINAL CHEMISTRY》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016181312A1 (en) * | 2015-05-11 | 2016-11-17 | Versitech Limited | Polycyclic epoxides and compositions thereof with anti-cancer activities |
| CN106749496A (en) * | 2016-12-05 | 2017-05-31 | 张奇军 | New triptolide derivative and its preparation and use |
| CN109021061A (en) * | 2018-09-29 | 2018-12-18 | 郭可点 | Triptolide targeted prodrug and its preparation method and application |
| CN109021061B (en) * | 2018-09-29 | 2019-07-12 | 郭可点 | Triptolide targeted prodrug and its preparation method and application |
| CN111821308A (en) * | 2019-04-15 | 2020-10-27 | 中国科学院上海药物研究所 | Application of tripterygium wilfordii in preparation of medicine for treating non-alcoholic fatty liver disease |
| CN111821308B (en) * | 2019-04-15 | 2021-10-08 | 中国科学院上海药物研究所 | Application of tripterygium wilfordii in preparation of medicine for treating non-alcoholic fatty liver disease |
| CN110511202A (en) * | 2019-08-13 | 2019-11-29 | 厦门大学 | Polycyclic polyisocyanate pentenyl acyl phloroglucinol class compound and the preparation method and application thereof |
| CN114478684A (en) * | 2022-01-21 | 2022-05-13 | 南京中医药大学 | Triptolide prodrug, preparation method and medical application thereof |
| WO2023138147A1 (en) * | 2022-01-21 | 2023-07-27 | 南京中医药大学 | Triptolide prodrug, preparation method therefor and pharmaceutical use thereof |
| CN114478684B (en) * | 2022-01-21 | 2023-10-31 | 南京中医药大学 | Tripterine prodrug, preparation method and medical application thereof |
| WO2023143330A1 (en) * | 2022-01-25 | 2023-08-03 | 杭州卫本医药科技有限公司 | Tripterygium wilfordii diterpene epoxide having function of killing myc positive cell, preparation method therefor, and application thereof |
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