CN103127054B - Applications of dihydro chromone framework compounds in preparing medicine for curing malignant tumor - Google Patents
Applications of dihydro chromone framework compounds in preparing medicine for curing malignant tumor Download PDFInfo
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Abstract
The invention discloses applications of dihydro chromone framework compounds of CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 in preparing medicine for curing malignant tumors. According to experimental proof, any kind of the six compounds can kill a plurality kinds of tumor cells of human bodies effectively, the tumor cells comprise cervical cancer cells, non-small cell lung cancer cells, breast cancer cells, ovarian carcinoma cells, hepatoma carcinoma cells, nasopharyngeal carcinoma cells, gastric carcinoma cells, laryngocarcinoma cells, pancreatic cancer cells, melanoma cells, bladder cancer cells and leukemia cells. Besides, with increase of dosages of the compounds, injured and killed cancers are increased with the increase of the dosages, and thus the new applications are proofed and discovered in the medicine for curing malignant tumors, and a new application range is exploited. Besides, new medicine is provided for the malignant tumors, and the difficulty in the chemical treatment of diseases that tumor cells are tolerant of chemotherapeutics is solved.
Description
Technical field
The present invention relates to the purposes of chromanone framework compound, the particularly application of chromanone framework compound in the medicine of preparation treatment malignant tumor.
Background technology
Malignant tumor is the major disease of harm humans health, has been first of China resident cause of the death.Chemotherapy is still the Main Means of many treating malignant tumors at present, but the tolerance of tumor cell to chemotherapeutics comprises primary drug resistance and acquired drug-resistance, is the matter of utmost importance that perplexs clinically and restrict Chemotherapy for Malignant Tumors always.Thereby, seek that to have the original new drug of efficient active anticancer very urgent.In recent years, natural product and derivant thereof have become the important sources of anticancer original new drug.No matter develop a kind new medicine by complete synthesis means, be for solving medicament sources, realizes the suitability for industrialized production of medicine, or all significant for resources and environment protection.
Patent CN102516217 discloses " chirality chromanone framework compound and asymmetric synthesis ", the method be under the condition of 0 DEG C～50 DEG C in organic solvent, taking chromone electron deficiency diolefin compound and olefine aldehydr as raw material, carry out the Diels-Alder reaction of antielectron demand or series connection Diels-Alder/ vinylogy Aldol, the reaction of series connection Diels-Alder/ hemiacetalization with Chiral Amine (particularly α, α-diaryl Prolinol silicon ether) as catalyst.The structural formula of one of products therefrom is as follows:
Wherein * is chiral carbon atom.R1 is selected from functional group's (for example ester group, cyano group, sulfuryl, nitro) with sucting electronic effect, R2, R3, R4, R5 are selected from H, the alkyl of C1-C16 or aryl, R6, R7, R8, R9 are selected from the oxyl of H, F, Cl, Br, C1-C16, alkyl or the aryl of C1-C16.But this patent does not disclose the biological activity of this compounds, more do not disclose its anti-tumor activity and to tumor cytotoxicity effect.
Summary of the invention
The object of the present invention is to provide the new purposes of chromanone framework compound, i.e. new purposes in pharmacy, for treatment malignant tumor provides novel drugs.
The present invention proves by experiment: chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23, any in CPQ-24 can effectively kill people's kinds of tumor cells, comprise cervical cancer cell, non-small cell lung cancer cell, breast cancer cell, ovarian cancer cell, hepatoma carcinoma cell, nasopharyngeal carcinoma cell, stomach cancer cell, laryngeal cancer cell, pancreatic cancer cell, melanoma cell, transitional cell bladder carcinoma cell line and leukaemia, and along with the increase of compound dosage, by killed and wounded with dead cell also dose dependent increase, its tumor killing cell effect presents dose-dependence.Thereby prove that chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 can apply in the medicine of preparation treatment malignant tumor.
Chromanone framework compound CPQ-16 of the present invention, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 meet disclosed following general structural formula in CN102516217:
In described general structure, work as R
1for CO
2et, R
2, R
4, R
5, R
6, R
7, R
9for H, R
3for
r
8during for F, be Compound C PQ-16, its structural formula is:
In described general structure, work as R
1for CO
2et, R
2, R
4, R
5, R
6, R
9for H, R
3for
r
7, R
8for Me(methyl) time, being compound is CPQ-17, its structural formula is:
In described general structure, work as R
1for CO
2et, R
2, R
4, R
5, R
6, R
7, R
9for H, R
3for
r
8during for F, be Compound C PQ-20, its structural formula is:
In described general structure, work as R
1for CO
2et, R
2, R
4, R
5, R
6, R
8, R
9for H, R
3for
r
7during for OAc, be Compound C PQ-22, its structural formula is:
In described general structure, work as R
1for CO
2et, R
2, R
4, R
5, R
7, R
9for H, R
3for
r
6, R
8during for F, be Compound C PQ-23, its structural formula is:
In described general structure, work as R
1for CO
2et, R
2, R
4, R
5, R
6, R
8, R
9for H, R
3for
r
7during for F, be Compound C PQ-24, its structural formula is:
The present invention is by adding respectively chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 are to after cultivating in Non-small cell lung carcinoma cell A549, under inverted phase contrast microscope, observe the morphological change of tumor cell, find that part cell volume dwindles, become circle, cell membrane shrinkage, depart from adjacent normal cell, Cytoplasm density increases, part cell has been cracked into cell debris, be suspended in culture fluid, prove these six kinds of chromanone framework compound energy Efficient killing effects and killed Non-small cell lung carcinoma cell A549.Killed and wounded simultaneously, dead lung cell A549 increases along with the increase of these six kinds of chromanone framework compound dosage dependency.
The present invention adopts lactic acid dehydrogenase release experiment to detect respectively chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the cytotoxicity of CPQ-24 to human cervical carcinoma cell, non-small cell lung cancer cell, breast cancer cell, ovarian cancer cell, hepatoma carcinoma cell, nasopharyngeal carcinoma cell, stomach cancer cell, laryngeal cancer cell, pancreatic cancer cell, melanoma cell, transitional cell bladder carcinoma cell line and leukaemia.Prove that these six kinds of chromanone framework compounds all can effectively kill above-mentioned tumor cell, and killed and wounded, dead cell increases along with the increase of these six kinds of chromanone framework compound dosage dependency, be dose dependent and kill and wound these human tumor cells.
In above-mentioned lactic acid dehydrogenase (being called for short LDH) release experiment, adopt lactic dehydrogenase enzyme detection kit (Cyto Tox
non-Radioactive Cytotoxicity Assay, Promega, article No. G1780, lot number 0000015327) detection death of neoplastic cells rate.Its ultimate principle is: LDH is the intracytoplasmic a kind of enzyme of living cells, in the time of cell injury, cell membrane fragmentation, LDH can be released in cell culture fluid, the LDH catalytic substrate discharging produces coloured compound, there is a high absworption peak at 490nm place, according to the OD value reading, can calculate cell mortality
Beneficial effect of the present invention:
1, the present invention has excavated new purposes to chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24, and the application in the medicine of preparation treatment malignant tumor, has opened up new application.
2, the present invention provides new active drug to human cervical carcinoma, nonsmall-cell lung cancer, breast carcinoma, ovarian cancer, hepatocarcinoma, nasopharyngeal carcinoma, gastric cancer, laryngeal carcinoma, cancer of pancreas, melanoma, bladder cancer and leukemic treatment, promote the development in treating malignant tumor field, there is very large Social benefit and economic benefit.
Brief description of the drawings
Fig. 1 non-small cell lung cancer cell A549 schematic diagram of morphological change after chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 process 24h respectively of behaving.(-) negative matched group in figure.
Detailed description of the invention
Embodiment 1: the preparation of chromanone framework compound CPQ-16
In reaction tube, add successively 0.02mmol hexahydropyridine catalyst, 0.1mmol chromone electron deficiency diolefin compound,
0.2mmol long-chain olefine aldehydr,
0.02mmol acidic additive o-fluorobenzoic acid and 1mL solvent 1,4-dioxane, under agitation react in normal pressure, 25 DEG C, TLC monitors reaction, after 12 hours, reaction finishes, decompression and solvent recovery, and residue obtains target product CPQ-16 by column chromatography for separation, yield 68%, structural formula is as follows:
1h NMR (400MHz, CDCl
3): δ=8.04 (dd, J=8.8Hz, J=6.4Hz, 1H), 7.19 (s, 1H), 6.82 (td, J=8.4Hz, J=2.4Hz, 1H), 6.67 (dd, J=10.0Hz, J=2.4Hz, 1H), 5.15 (s, 1H), 4.57 (s, 1H), 4.38 (dd, J=10.0Hz, J=2.8Hz, 1H), 4.21 (q, J=7.2Hz, 2H), 2.27 (dd, J=13.6Hz, J=8.0Hz, 1H), 2.12-2.05 (m, 1H), 1.97-1.94 (m, 4H), 1.70 (s, 3H), 1.61 (s, 3H), 1.57-1.53 (m, 1H), 1.44-1.34 (m, 1H), 1.30 (t, J=7.2Hz, 3H) ppm,
13c NMR (100MHz, CDCl
3): δ=191.4,167.5,165.2,162.7,143.3,133.5,131.7,130.6,124.1,116.7,110.6,104.7,78.5,67.4,60.9,55.0,44.5,41.0,37.9,34.9,25.7,23.8,17.7,14.2ppm, high resolution mass spectrum calculating value: C
24h
27fO
5+ Na437.1740, measured value: 437.1740.
Embodiment 2: the preparation of chromanone framework compound CPQ-17
In reaction tube, add successively 0.02mmol hexahydropyridine catalyst, 0.1mmol chromone electron deficiency diolefin compound,
0.2mmol long-chain olefine aldehydr,
0.02mmol acidic additive o-fluorobenzoic acid and 1mL solvent 1,4-dioxane, under agitation react in normal pressure, 25 DEG C, TLC monitors reaction, after 12 hours, reaction finishes, decompression and solvent recovery, and residue obtains target product CPQ-17 by column chromatography for separation, yield 73%, structural formula is as follows:
1h NMR (400MHz, CDCl
3): δ=7.74 (s, 1H), 7.20 (s, 1H); 6.76 (s, 1H), 5.16-5.14 (m, 1H); 4.56 (dd, J=8.0Hz, J=1.6Hz, 1H); 4.31 (dd, J=10.0Hz, J=3.2Hz, 1H); 4.21 (q, J=7.2Hz, 2H); 2.28 (s, 3H), 2.24-2.22 (m; 4H), 2.09-1.92 (m, 5H); 1.70 (s, 3H), 1.61 (s; 3H), 1.55 (dd, J=13.2Hz; J=3.2Hz, 1H), 1.40 (dt; J=13.6Hz, J=2.8Hz, 1H); 1.32 (t, J=7.2Hz, 3H) ppm;
13cNMR (100MHz, CDCl
3): δ=192.8,165.3,159.5,146.9,142.8,134.4,131.5,130.8,127.9,124.3,118.3,117.5,77.9,67.5,60.7,55.2,44.3,41.0,38.0,34.9,25.7,23.8,20.5,18.8,17.6,14.1ppm; High resolution mass spectrum calculating value: C
26h
32o
5+ Na447.2147, measured value: 447.2149.
Embodiment 3: the preparation of chromanone framework compound CPQ-20
In reaction tube, add successively 0.02mmol hexahydropyridine catalyst, 0.1mmol chromone electron deficiency diolefin compound,
0.2mmol long-chain olefine aldehydr,
0.02mmol acidic additive o-fluorobenzoic acid and 1mL solvent 1,4-dioxane, under agitation react in normal pressure, 25 DEG C, TLC monitors reaction, after 12 hours, reaction finishes, decompression and solvent recovery, and residue obtains target product CPQ-20 by column chromatography for separation, yield 91%, structural formula is as follows:
1h NMR (400MHz, CDCl
3): δ=8.02 (dd, J=8.8Hz, J=6.4Hz, 1H); 7.18 (s, 1H), 6.81 (td, J=8.4Hz; J=2.4Hz, 1H), 6.66 (dd, J=9.6Hz; J=2.0Hz, 1H), 5.16-5.09 (m, 2H); 4.56 (s, 1H), 4.37 (dd, J=10.4Hz; J=2.8Hz, 1H), 4.20 (m, 2H); 2.30-2.24 (m, 1H), 2.10-1.96 (m, 9H); 1.68 (s, 3H), 1.60-1.54 (m; 7H), 1.43-1.38 (m, 1H); 1.31 (t, J=7.2Hz, 3H) ppm;
13c NMR (100MHz, CDCl
3): δ=191.4,167.6,165.3,162.7,143.3,135.3,133.5,131.4,130.6,124.8; 124.3,116.7,110.5,104.8,78.5,67.4,60.9,55.0,44.5,41.1; 40.0,37.9,34.9,32.0,26.7,25.7,23.7,17.7,14.2ppm; High resolution mass spectrum calculating value: C
29h
35fO
5+ Na505.2366, measured value: 505.2367.
Embodiment 4: the preparation of chromanone framework compound CPQ-22
In reaction tube, add successively 0.02mmol hexahydropyridine catalyst, 0.1mmol chromone electron deficiency diolefin compound,
0.2mmol long-chain olefine aldehydr,
0.02mmol acidic additive o-fluorobenzoic acid and 1mL solvent 1,4-dioxane, under agitation react in normal pressure, 25 DEG C, TLC monitors reaction, after 12 hours, reaction finishes, decompression and solvent recovery, and residue obtains target product CPQ-22 by column chromatography for separation, yield 51%, structural formula is as follows:
1h NMR (400MHz, CDCl
3): δ=8.04 (d, J=8.8Hz, 1H), 7.20 (s, 1H), 6.84 (dd, J=8.8Hz, J=2.0Hz, 1H), 6.77 (d, J=2.0Hz, 1H), 5.15 (s, 1H), 4.59 (d, J=7.2Hz, 1H), 4.39 (dd, J=10.0Hz, J=3.2Hz, 1H), 4.22 (q, J=7.2Hz, 2H), 2.33 (s, 3H), 2.27 (dd, J=13.6Hz, J=8.4Hz, 1H), 2.12-2.05 (m, 1H), 2.01-1.92 (m, 4H), 1.70 (s, 3H), 1.61-1.56 (m, 4H), 1.42 (dt, J=13.6Hz, J=2.8Hz, 1H), 1.33 (t, J=7.2Hz, 3H) ppm,
13c NMR (100MHz, CDCl
3): δ=191.8,168.5,165.3,162.0,156.6,143.1,133.3,131.6,129.5,123.8,117.7,116.2,110.9,78.4,67.3,60.8,55.1,44.4,41.0,37.9,34.9,25.7,23.8,21.1,17.7,14.1ppm, high resolution mass spectrum calculating value: C
26h
30o
7+ K493.1629, measured value: 493.1638.
Embodiment 5: the preparation of chromanone framework compound CPQ-23
In reaction tube, add successively 0.02mmol hexahydropyridine catalyst, 0.1mmol chromone electron deficiency diolefin compound,
0.2mmol long-chain olefine aldehydr,
0.02mmol acidic additive o-fluorobenzoic acid and 1mL solvent 1,4-dioxane, under agitation react in normal pressure, 25 DEG C, TLC monitors reaction, after 12 hours, reaction finishes, decompression and solvent recovery, and residue obtains target product CPQ-23 by column chromatography for separation, yield 75%, structural formula is as follows:
1h NMR (400MHz, CDCl
3): δ=7.49 (m, 1H), 7.16-7.10 (m; 2H), 5.16-5.09 (m, 2H); 4.61 (s, 1H), 4.42 (dd; J=10.0Hz, J=2.4Hz, 1H); 4.25-4.19 (m, 2H), 2.36-2.30 (m; 1H), 2.15-1.94 (m, 9H); 1.74-1.65 (m, 7H), 1.61 (s; 3H); 1.42 (m, 1H), 1.33 (t; J=7.2Hz, 3H) ppm;
13c NMR (100MHz, CDCl
3): δ=190.8,165.2,156.1,151.4,143.7,135.4,132.8,131.4,124.7,124.2; 123.9,121.6,111.1,108.3,79.2,67.2,61.0,55.5,44.5,41.1; 39.6,37.8,34.8,31.9,26.6,25.7,23.6,17.7,14.1ppm; High resolution mass spectrum calculating value: C
29h
34f
2o
5+ Na523.2272, measured value: 523.2272.
Embodiment 6: the preparation of chromanone framework compound CPQ-24
In reaction tube, add successively 0.02mmol hexahydropyridine catalyst, 0.1mmol chromone electron deficiency diolefin compound,
0.2mmol long-chain olefine aldehydr,
0.02mmol acidic additive o-fluorobenzoic acid and 1mL solvent 1,4-dioxane, under agitation react in normal pressure, 25 DEG C, TLC monitors reaction, after 12 hours, reaction finishes, decompression and solvent recovery, and residue obtains target product CPQ-24 by column chromatography for separation, yield 43%, structural formula is as follows:
1h NMR (400MHz, CDCl
3): δ=8.04 (dd, J=8.8Hz, J=6.4Hz, 1H); 7.20 (s, 1H), 6.82 (td, J=8.4Hz; J=2.4Hz, 1H), 6.67 (dd, J=9.6Hz; J=2.4Hz, 1H), 5.17-5.10 (m, 2H); 4.57 (d, J=7.2Hz, 1H); 4.39 (dd, J=9.6Hz, J=2.4Hz; 1H), 4.21 (m, 2H); 2.32-2.25 (m, 1H), 2.12-1.98 (m; 9H), 1.69 (s, 3H); 1.61 (s, 6H), 1.57 (m; 1H), 1.42 (m, 1H); 1.32 (t, J=7.2Hz, 3H) ppm;
13c NMR (100MHz, CDCl
3): δ=191.4,167.6,165.3,162.7,143.3,135.3,133.5,131.4,130.6,124.8; 124.3,116.7,110.5,104.8,78.5,67.4,60.9,55.0,44.5,41.1; 40.0,37.9,34.9,32.0,26.7,25.7,23.7,17.7,14.2ppm; High resolution mass spectrum calculating value: C
29h
35fO
5+ Na505.2366, measured value: 505.2369.
In the detection embodiment of following chromanone framework compound anti-tumor activity, the PRMI1640 culture fluid of described 10% hyclone is purchased from Hyclone Thermo Scientific company, specification 500ml/ bottle, lot number NXK0719; Described inverted phase contrast microscope is Olympus(Olympus), CKX41 model; Described lactic dehydrogenase enzyme detection kit is Cyto Tox
non-Radioactive Cytotoxicity Assay, Promega, article No. G1780, lot number 0000015327; Described microplate reader is Infinite M2000 global function microplate reader, TECAN; Described tumor cell line is all purchased from the biological product collecting center of Unite States Standard (American Type Culture Collection, ATCC).
7: six kinds of chromanone framework compound CPQ-16 of embodiment, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the killing experiments of CPQ-24 to Non-small cell lung carcinoma cell A549.
1, experimental technique
With the PRMI1640 culture fluid containing 10% hyclone, 37 DEG C of juxtapositions, 5%CO
2in incubator, cultivate Non-small cell lung carcinoma cell A549 cell, within every 3 days, go down to posterity with 0.25% trypsin solution peptic cell, and change culture fluid.CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 are used respectively to DMSO(dimethyl sulfoxide) be made into the storage liquid of 20mM.A549 cell is pressed to 0.7 × 10
4/ hole concentration is inoculated on 96 orifice plates, cultivates after 24h, adds respectively CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24, and the final concentration of every kind of compound adds according to 2.5 μ M, 5 μ M, 7.5 μ M, 10 μ M.The DMSO final concentration (V/V) adding due to above-mentioned four kinds of concentration is respectively 0.0125%, 0.025%, 0.0375%, 0.05%, and for getting rid of the lethal effect of solvent DMSO to cell, negative control group cell adds the DMSO that final concentration (V/V) is 0.05%.Establish four multiple holes, then 96 orifice plates are put to 37 DEG C, 5%CO for each group
2incubator is cultivated 24h again.
2, fragmentation effect is observed: Tissue Culture Plate is placed in and under inverted phase contrast microscope, observes tumor cell morphological change, and take a picture, negative control group, CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 final concentration are that Fig. 1 is shown in by the picture of 5 μ M, 10 μ M.As shown in Figure 1, negative control group A549 cell attachment well-grown, has no dead floating cell.The A549 cell partial volume of six kinds of chromanone framework compound processing 24h dwindles, becomes circle, and cell membrane shrinkage departs from adjacent normal cell, and Cytoplasm density increases, and part cell has been cracked into cell debris, is suspended in culture fluid.Can find out simultaneously, along with the increase of compound dosage, be killed and wounded, dead cell also increases dose dependent, experimental result shows, six kinds of chromanone framework compounds all can efficiently kill A549 cell, and lethal effect presents dose-dependence.
Embodiment 8: adopt lactic acid dehydrogenase release experiment, detect chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the lethal effect of CPQ-24 to Non-small cell lung carcinoma cell A549.
1, experimental technique
The method conservation that A549 cell is pressed in embodiment 7 is cultivated, by 0.7 × 10
4/ hole concentration is inoculated on 96 orifice plates, puts 37 DEG C, 5%CO
2in incubator, cultivate after 24h, add respectively Compound C PQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24, every kind of compound adds according to final concentration 0.625 μ M, 1.25 μ M, 2.5 μ M, 5 μ M, 7.5 μ M, 10 μ M, 15 μ M, 20 μ M, negative control group cell adds 0.1%(v/v) DMSO(and compound concentration for the DMSO concentration that the experimental group of 20 μ M adds identical), separately establish zeroing group (be there is no cell in cell culture hole, only add cell culture fluid).Establish four multiple holes, then 96 orifice plates are put to 37 DEG C, 5%CO for each group
2incubator is cultivated 72h again, then the lethal effect of detection compound to tumor cell as follows.
2, detection method:
By lactic acid dehydrogenase (LDH) release experiment, adopt lactic dehydrogenase enzyme detection kit to detect death of neoplastic cells rate, concrete operation method carries out according to the description of described lactic dehydrogenase enzyme detection kit.With detecting the resuspended reaction substrate of buffer (Assay Buffer) (Substrate Mix).Get in 96 porocyte culture plates in each group l to the 96 new orifice plates of culture supernatant 50 μ in 3 multiple holes, it is 0.9%(V/V that a remaining multiple hole of each group adds final concentration) Triton-x100 (test kit provides), and be placed in 37 DEG C of incubators 50 minutes with after cell lysis, getting 50 μ l supernatant adds in 96 new orifice plates, then in 96 new orifice plates, every hole adds isopyknic resuspended good reaction substrate liquid again, after incubated at room 30 minutes, it is acetic acid (providing in the test kit) cessation reaction of 1M that every hole adds 50 μ l stop buffers, survey the OD value in every hole by microplate reader (490nm wavelength), the OD value that cell lysis hole is detected is designated as " the maximum OD of release of cell value ", the OD value of negative control group is designated as " Spontaneous release matched group OD value ".
According to the OD490 value detecting, calculate according to the following formula cell mortality, result represents with mean ± standard deviation, and adopts SPSS software Pro bit module calculating half to kill cell concentration, i.e. IC50.
3, testing result:
As shown in table 1, these six kinds of compounds all can effectively kill A549 cell, and along with the increase of compound concentration, they to the lethal effect of A549 also dose dependent increase, by SPSS software Pro bit module calculate, CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 cause the effective dose (IC50) of half A549 cell death to be respectively 7.2 μ M, 9.3 μ M, 8.2 μ M, 7.2 μ M, 4.0 μ M and 4.9 μ M.
Table 1.CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the external Mortaility results of CPQ-24 to A549 cell
Embodiment 9: adopt lactic acid dehydrogenase release experiment, detect chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the lethal effect of CPQ-24 to human cervical carcinoma cell HeLa.
According to the cell culture processes in embodiment 7, human cervical carcinoma cell HeLa cell is cultivated and is inoculated in 96 well culture plates by conservation.Establish experimental group, matched group and zeroing group by the method in embodiment 8, and by lactic acid dehydrogenase (LDH) release experiment, detect respectively the HeLa cell mortality that CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 cause, result represents with mean ± standard deviation, and calculates the dosage (IC that causes half HeLa cell death
50).
Testing result: as shown in table 2, these six kinds of compounds all can effectively kill HeLa cell, and along with the increase of compound concentration, they to the lethal effect of HeLa also dose dependent increase, by SPSS software Pro bit module calculate, CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 cause the effective dose of half HeLa cell death to be respectively 5.5 μ M, 7.1 μ M, 5.8 μ M, 6.2 μ M, 2.9 μ M and 4.2 μ M.
Table 2.CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the external Mortaility results of CPQ-24 to HeLa cell
Embodiment 10: adopt lactic acid dehydrogenase release experiment, detect chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24 to multiple human tumor cell, comprise breast carcinoma, ovarian cancer, hepatocarcinoma, nasopharyngeal carcinoma, gastric cancer, laryngeal carcinoma, cancer of pancreas, melanoma, bladder cancer and leukaemia's lethal effect.
The method conservation of pressing in embodiment 7 is cultivated breast cancer cell (MCF-7), ovarian cancer cell (SKOV3), hepatoma carcinoma cell (Bel-7402), nasopharyngeal carcinoma cell (HNE), stomach cancer cell (MKN-45), laryngeal cancer cell (Hep-2), pancreatic cancer cell (Pan-1), melanoma cell (A375), transitional cell bladder carcinoma cell line (Biu-87).The same employing of leukaemia (Jurkat) contains 10% hyclone PRMI1640, is placed in 37 DEG C, 5%CO
2in incubator, cultivate, and suspension cell culture mode goes down to posterity routinely, in the time going down to posterity, first do centrifugal treating and remove old culture fluid, then add new culture fluid.The tumor cell of cultivating is pressed to 0.7 × 10
4/ hole concentration is inoculated in 96 orifice plates, cultivate after 24h, according to every kind of compound 2.5 μ M, the final concentration of 5 μ M and 10 μ M adds six kinds of chromanone framework compound CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and CPQ-24, continue to cultivate 72h, then establish experimental group, matched group and zeroing group by embodiment 8 methods, and detect death of neoplastic cells rate by lactic acid dehydrogenase (LDH) release experiment, result represents with mean ± standard deviation.
Testing result: kill and wound these human tumor cells, as shown in Table 3 and Table 4, they have very strong antitumor action as seen above-mentioned six kinds of chromanone framework compound dose dependents.
Table 3. final concentration is that CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the CPQ-24 of 5 μ M are to the external Mortaility results of various human tumor cell
Table 4. final concentration is that CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23 and the CPQ-24 of 10 μ M are to the external Mortaility results of various human tumor cell
Claims (2)
1. the application of chromanone framework compound in the medicine of preparation treatment malignant tumor, described chromanone framework compound is the one in CPQ-16, CPQ-17, CPQ-20, CPQ-22, CPQ-23, CPQ-24,
The structural formula of described CPQ-16 is
The structural formula of described CPQ-17 is
The structural formula of described CPQ-20 is
The structural formula of described CPQ-22 is
The structural formula of described CPQ-23 is
The structural formula of described CPQ-24 is
2. application according to claim 1, is characterized in that described malignant tumor is the one in cervical cancer, nonsmall-cell lung cancer, breast carcinoma, ovarian cancer, hepatocarcinoma, nasopharyngeal carcinoma, gastric cancer, laryngeal carcinoma, cancer of pancreas, melanoma, bladder cancer, leukemia.
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| WO2009108392A2 (en) * | 2008-02-29 | 2009-09-03 | Northwestern University | Catalytic enantioselective synthesis of flavanones and chromanes |
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| CN102516217A (en) * | 2011-12-15 | 2012-06-27 | 四川大学 | Compound containing chiral chromanone skeletons and asymmetric synthetic method thereof |
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| Title |
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| JP特开平9-301915A 1997.11.25 |
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| 应华州.含氮姜黄素和类黄酮衍生物的设计、合成和抗肿瘤活性研究.《中国博士论文全文数据库》.2009,全文. |
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