Summary of the invention
The present invention is directed to overcome the shortage of prior art, a kind of alkaloid for reversing lung cancer cisplatin-resistant is provided.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
As the alkaloid of flowering structure is preparing the application in the drug for reversing patients with lung cancer cisplatin-resistant:
Wherein, R is-CH3Or-CH2CH2CH3。
It is a kind of reverse patients with lung cancer cisplatin-resistant pharmaceutical preparation also contain medicine using above-mentioned alkaloid as active constituent
Acceptable auxiliary material on.
As the alkaloid of flowering structure is preparing the application in the drug for reversing patients with lung cancer cisplatin-resistant:
Wherein, R is
It is a kind of reverse patients with lung cancer cisplatin-resistant pharmaceutical preparation also contain medicine using above-mentioned alkaloid as active constituent
Acceptable auxiliary material on.
As the alkaloid of flowering structure is preparing the application in the drug for reversing patients with lung cancer cisplatin-resistant:
Wherein, R is
It is a kind of reverse patients with lung cancer cisplatin-resistant pharmaceutical preparation also contain medicine using above-mentioned alkaloid as active constituent
Acceptable auxiliary material on.
As the alkaloid of flowering structure is preparing the application in the drug for reversing patients with lung cancer cisplatin-resistant:
Wherein, R is
It is a kind of reverse patients with lung cancer cisplatin-resistant pharmaceutical preparation also contain medicine using above-mentioned alkaloid as active constituent
Acceptable auxiliary material on.
As the alkaloid of flowering structure is preparing the application in the drug for reversing patients with lung cancer cisplatin-resistant:
Wherein, R is
It is a kind of reverse patients with lung cancer cisplatin-resistant pharmaceutical preparation also contain medicine using above-mentioned alkaloid as active constituent
Acceptable auxiliary material on.
The utility model has the advantages that
It is a discovery of the invention that compound TA-1, TA-2, TB-1, TB-2, TB-3, TC-1, TD-1, TE-1, TE-2 can be effective
Reverse lung carcinoma cell to the drug resistance of cis-platinum, wherein TC-1, TD-1 are not only only capable of reversing drug resistance, additionally it is possible to enhance lung carcinoma cell
To the sensibility of cis-platinum.Compound TB-4 does not have this effect.
Specific embodiment
It is specific with reference to the accompanying drawings and examples to introduce essentiality content of the present invention, but guarantor of the invention cannot be limited with this
Protect range.
One, experimental material
The alkaloid for studying test is prepared with literature method according to a conventional method, is divided into five major class according to chemical structure, specifically
Chemical structure and number are as shown in the table.
Lung cancer cell line H460 is purchased from ATCC, and DMEM culture medium, fetal calf serum are purchased from GIBCO company.
Two, experimental method
1, cell culture
Human lung carcinoma cell line H460 is routinely after recovery, with containing 10%FBS, 50U/ml streptomysin, 50U/ml penicillin and
The DMEM culture medium of 4mmol/L L-Glutamine, in 37 DEG C, 5%CO2Under the conditions of cultivate, every 2~3d changes liquid.
2, cisplatin resistance strain H460/DDP is constructed
It is incremented by the human lung cancer cisplatin-resistant cell strain H460/DDP that revulsion is established with cisplatin continuous exposure concentration.
The specific method is as follows: the H460 cell of logarithmic growth phase is cultivated with the DMEM that cis-platin concentrations are 0.1 μ g/ml, after 4 weeks
Cell dissociation is passed on to and used normal full training culture, after cell is adherent, cis-platin concentrations is improved to 0.2 μ g/ml, continue to cultivate
4 weeks and had digestive transfer culture;Successively cis-platin concentrations are improved to 0.5,1.0,2.0 μ g/ml again, it is suitable to obtain lung cancer for continuous culture 5 months
Platinum persister H460/DDP.
3, it is grouped and is administered
Administration group: the H460/DDP cell of logarithmic growth phase is inoculated in 96 orifice plates, 37 DEG C, 5% with 1000/hole
CO2It is cultivated in incubator.After cell is adherent, be separately added into containing 5 μM of TA-1, TA-2, TB-1, TB-2, TB-3, TB-4,
The complete medium culture of TC-1, TD-1, TE-1 or TE-2 (DMSO is solvent);
H460/DDP group: equivalent solvent DMSO, not dosing, other same administration groups are only added in H460/DDP cell;
H460 group: equivalent solvent DMSO, not dosing, other same administration groups are only added in H460 cell.
4, measurement of the group of cells to cisplatin sensitivity
After group of cells culture 48h, old culture medium is discarded, the cis-platinum of gradient concentration is added, each concentration is arranged 3 again
Hole, 37 DEG C, 5%CO2After cultivating 48h in incubator, old culture medium is abandoned, the fresh culture containing 10%CCK-8 is added, 37 DEG C incubate
3h is educated, OD value at 450nm, calculation of half inhibitory concentration IC50 are surveyed.
5, data processing
Statistical analysis is carried out using SPSS 17.0, data are indicated using means standard deviation, using Student ' s t
It examines and carries out two comparison among groups, P < 0.05 is that difference is statistically significant.
Three, experimental result
Cis-platinum is to group of cells half-inhibitory concentration IC50 value as shown in table 1 and Fig. 1.Compared with H460 group, H460/DDP
The IC50 value of group lung carcinoma cell significantly increases (P < 0.05), and it is apparent resistance to illustrate that H460/DDP lung carcinoma cell produces cis-platinum
Pharmacological property;Compared with H460/DDP group, TA-1, TA-2, TB-1, TB-2, TB-3, TC-1, TD-1, TE-1, TE-2 group lung carcinoma cell
IC50 value significantly reduce (P < 0.05), illustrate that TA-1, TA-2, TB-1, TB-2, TB-3, TC-1, TD-1, TE-1, TE-2 are incubated
Educate the sensibility (P < 0.05) for enhancing H460/DDP lung carcinoma cell to cis-platinum.
Compound TB-4 acts on unobvious (P > 0.05).
1 cis-platinum of table is to group of cells half-inhibitory concentration IC50 value
Group |
IC50 value (μ g/ml) |
Group |
IC50 value (μ g/ml) |
H460 group |
14.21±1.25 |
TB-3 administration group |
20.06±1.19 |
H460/DDP group |
52.85±1.63 |
TB-4 administration group |
48.72±1.55 |
TA-1 administration group |
21.07±1.47 |
TC-1 administration group |
7.61±0.93 |
TA-2 administration group |
23.42±1.38 |
TD-1 administration group |
7.33±0.88 |
TB-1 administration group |
18.95±1.10 |
TE-1 administration group |
15.62±1.27 |
TB-2 administration group |
17.18±1.08 |
TE-2 administration group |
9.24±0.96 |
It is above-mentioned the experimental results showed that, compound TA-1, TA-2, TB-1, TB-2, TB-3, TC-1, TD-1, TE-1, TE-2 can
Effectively to reverse lung carcinoma cell to the drug resistance of cis-platinum, wherein TC-1, TD-1 are not only only capable of reversing drug resistance, additionally it is possible to enhance lung
Sensibility of the cancer cell to cis-platinum.Compound TB-4 does not have this effect.
The effect of above-described embodiment is specifically to introduce essentiality content of the invention, but cannot be by protection of the invention
Range is confined to above-mentioned specific embodiment.