CN103127101A - Application of 8-diethyl malonate berberine in preparation of antitumor drug - Google Patents
Application of 8-diethyl malonate berberine in preparation of antitumor drug Download PDFInfo
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- CN103127101A CN103127101A CN2011103750311A CN201110375031A CN103127101A CN 103127101 A CN103127101 A CN 103127101A CN 2011103750311 A CN2011103750311 A CN 2011103750311A CN 201110375031 A CN201110375031 A CN 201110375031A CN 103127101 A CN103127101 A CN 103127101A
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- diethyl malonate
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Abstract
The invention belongs to the chemical industry field and the medicine field, and relates to an application of 8-diethyl malonate berberine in preparation of antitumor drugs. The invention provides the application of 8-diethyl malonate berberine in preparation of antitumor drugs, and the tumor cells comprise leukemia cells and pancreas cancer cells. The 8-diethyl malonate berberine belongs to a natural product, has little toxic and side effect, high bioavailability, and stable properties, and has clinical application value. The micromolecular compound of the invention can be developed as a new antitumor drug or its auxiliary component, has obvious tumor suppression effect, is green and environment-friendly, and provides a new approach and means for treating and curing tumors.
Description
Technical field
The invention belongs to chemical field and field of medicaments, relate to the application of 8-diethyl malonate berberine in the preparation antitumor drug.
Background technology
8-diethyl malonate berberine is that 8,8-two replaces-13,13a-dihydro berberine derivant, and its structural formula is as follows:
This compound be seen in first in 1979 report (Chemistry of 8-chloroberberine.Journal ofPharmaceutical Sciences (1979), 68 (6), 705-8.).
application number is 200910049688.1, name is called " 8, 8-two replaces-13, 13a-dihydro berberine derivant and pharmaceutical composition thereof and purposes " Chinese invention patent application the synthesis preparation method of 8-diethyl malonate berberine is disclosed, and show that by experiment this compounds has the effect that promotes glucose absorption to muscle cell, the whole animal test shows that such compound has the carbohydrate tolerance of improvement and insulin resistant, alleviate obesity, alleviate the effects such as fatty liver, and bioavailability is high, strengthened the drug effect in body, while Stability Analysis of Structures under acid condition.This compounds can be used for treating type 2 diabetes mellitus, obesity, fatty liver or its complication.
8-diethyl malonate berberine is natural product, and side effect is little, bioavailability is high, stable in properties, has clinical use value.But, up to now, not yet see the report about the relevant anti-tumor aspect of this compound.
Summary of the invention
The new medicinal usage that the purpose of this invention is to provide 8-diethyl malonate berberine.
The invention provides the application of 8-diethyl malonate berberine in the preparation antitumor drug.Described tumor can be hepatocarcinoma, leukemia, adenocarcinoma of lung or cancer of pancreas.Described antitumor drug is injection or tablet.Described antitumor drug can be leukemia or cancer of pancreas medicine.
The invention provides a kind of method that suppresses tumor cell in vitro propagation, 8-diethyl malonate berberine is added in the culture fluid of tumor cell.Described tumor cell is hepatoma carcinoma cell, leukaemia, lung adenocarcinoma cell or pancreatic cancer cell.Generally speaking, adding the final concentration of 8-diethyl malonate berberine is 1-100 μ M.For example, 1-5 μ M, 1-10 μ M, 1-20 μ M, 1-30 μ M, 1-50 μ M, 3-60 μ M, 3-50 μ M, 3-30 μ M, 3-20 μ M, 3-10 μ M, 3-5 μ M, 5-20 μ M, etc.
The present invention also provides a kind of antitumor drug, and the active component of described antitumor drug is 8-diethyl malonate berberine.Described tumor can be hepatocarcinoma, leukemia, adenocarcinoma of lung or cancer of pancreas.
Micromolecular compound of the present invention can adopt the preparation method preparation of various routines.For example, adopt the synthetic method of artificial chemistry.
Utilize micromolecular compound of the present invention, by various conventional screening techniques, can filter out and interactional material occurs 8-diethyl malonate berberine, as receptor, inhibitor or pick up anti-dose etc.
The present invention and inhibitor thereof, pick up anti-dose etc., when using (administration) in treatment, can provide different effects.Usually, these materials can be formulated in nontoxic, inertia with pharmaceutically acceptable aqueous carrier medium in, wherein pH is about 5-8 usually, preferably pH is about 6-8, pH value can change to some extent with the character that is formulated material and disease to be treated.The pharmaceutical composition for preparing can carry out administration by conventional route, comprising (but being not limited to): intramuscular, intraperitoneal, subcutaneous, Intradermal or topical.
Take 8-diethyl malonate berberine of the present invention as example, can be with itself and suitable pharmaceutically acceptable carrier coupling.This class pharmaceutical composition contains compound and pharmaceutically acceptable carrier or the excipient for the treatment of effective dose.This class carrier comprises (but being not limited to): saline, buffer, glucose, water, glycerol, ethanol and combination thereof.Pharmaceutical preparation should be complementary with administering mode.8-diethyl malonate berberine of the present invention can be made into the injection form, for example is prepared by conventional method with normal saline or the aqueous solution that contains glucose and other adjuvant.Pharmaceutical composition such as Tablet and Capsula can be prepared by conventional method.Pharmaceutical composition such as injection, solution, Tablet and Capsula should be made under aseptic condition.The dosage of active component is the treatment effective dose, for example every day about 5 mg/kg body weight of 1 microgram/kg body weight-Yue.In addition, 8-diethyl malonate berberine of the present invention also can use together with the other treatment agent.
When 8-diethyl malonate berberine of the present invention is used as medicine, the 8-diethyl malonate berberine for the treatment of effective dose can be applied to mammal, wherein should treat effective dose usually at least about 10 micrograms/kg body weight, and in most of the cases be no more than approximately 8 mg/kg body weight, preferably this dosage is the about 1 mg/kg body weight of 10 micrograms/kg body weight-Yue.Certainly, concrete dosage also should be considered the factors such as route of administration, patient health situation, and these are all within the skilled practitioners skill.
The invention provides the application of 8-diethyl malonate berberine in the preparation antitumor drug.8-diethyl malonate berberine is natural product, obviously the propagation of inhibition tumor cell.Micromolecular compound of the present invention is developed as new antitumor drug or its auxiliary element, and tumor killing effect is obvious, environmental protection, and will and cure tumor for treatment provides a kind of new approach and means.
The specific embodiment
Experimental technique:
1. cell recovery
1) take out cryopreservation tube from liquid nitrogen container, directly drop in 37 ℃ of warm water, and frequently shake and make it melt as early as possible.
2) take out cryopreservation tube from 37 ℃ of water-baths, with suction pipe sucking-off cell suspension, inject centrifuge tube and add culture fluid more than 10 times, low-speed centrifugal after mixing is abandoned supernatant, then is repeated to wash once with culture fluid.
3) with after the suitable dilution of culture fluid, the inoculated and cultured bottle is placed on 37 ℃ of standing cultivations of incubator, changes culture fluid next day, continues to cultivate.Go down to posterity when being cultured to finite concentration.The PANC-1 cell culture is in containing the DMEM high glucose medium of 10%Gibico hyclone, and the K562 cell culture contains 100U/ml penicillin and 100 μ g/ml streptomycins in culture medium in containing 1640 culture medium of 10% hyclone.
2. passage is cultivated
The situation of observation of cell growth every day is grown to approximately 90% and is gone down to posterity when converging (attached cell) in culture bottle when cell, approximately went down to posterity once every 2-4 days.One bottle goes down to posterity into three bottles, or a 25cm
2Go down to posterity in a 75cm
2Culture bottle in.Method:
1) with 1 * phosphate buffer washed cell once.
2) add 2-3ml trypsinization liquid digestion, be placed in 37 ℃ of incubator numbers minute.Pat Tissue Culture Flask with hands, make cell separation.
3) suitable culture medium with the Gibico hyclone that contains 10-15% stops trypsinization.Cell is sub-packed in new culture bottle, continues to cultivate.
3. cell cryopreservation
1) get the cell tryptase protease digestion that is cultured to exponential phase, be collected in centrifuge tube and counting, centrifugal.
2) reject trypsin and old culture fluid add the frozen culture fluid (containing 10%DMSO, 40%DMEM and 50%Gibico hyclone) that configures, and in cryopreserving liquid, the ultimate density of cell is 0.5-1 * 10
7/ ml.Blow and beat gently with suction pipe and make cell even, then be distributed in aseptic cryopreservation tube, every pipe adds 1-1.5ml.
3) cryopreservation tube is put into freezing storing box and put-80 ℃ of quick-freezings, move in liquid nitrogen container after 5 hours and preserve.
The growth inhibited effect of embodiment 18-diethyl malonate berberine to human pancreatic cancer cell
PANC-1 cell (available from Chinese Academy of Sciences's cell bank) 3 * 10
3/ hole is seeded to 96 orifice plates, cultivates to make it in 24 hours to add 8-diethyl malonate berberine (available from Shanghai Pharmaceutical Inst., Chinese Academy of Sciences) after adherent, establishes 6 Concentraton gradient, and each concentration is established 3 multiple holes.Cell is at 37 ℃, 5%CO
2Cultivate after 72 hours under condition, outwell culture fluid, measure cell survival rate with MTS test kit (Promega company).
Method of testing is: cell is washed one time with serum-free medium, added the MTS chromophoric solution (adding 2ml solution 1 and 100 μ l solution 2 in the 10ml serum-free medium, fully mixing) for preparing in advance according to the amount of 100 μ l/well.Do not have the hole of cell to be made as this bottom outlet with one, absorb in order to the bias light of calibration solution.Cell is put into cell culture incubator to be continued to cultivate 2～4 hours, then read absorbance value (reference wavelength 630-700nm with microplate reader, measure wavelength 490nm), calculate cell survival rate, to measure hole absorbance value/control wells absorbance value as the numerical value of cell survival rate.According to cell survival rate, calculate 8-diethyl malonate berberine to the IC50 value of HGC cell.
IC50 refers to the concentration of a suppressed half inhibitor.Here be the PANC-1 cell quantity and be the concentration of a half 8-diethyl malonate berberine of contrast.The calculating of IC50 generally need to be measured the dosage effect more than 5, then obtains function calculation by curve fitting and get.
Result: 8-diethyl malonate berberine is 3.79 μ M to the IC50 value of PANC-1 cell.
The growth inhibited effect of embodiment 28-diethyl malonate berberine to the human leukemia cell
Utilize cck-8 test kit (Japanese colleague's chemistry institute) to detect.
Step:
1) K562 cell (available from Chinese Academy of Sciences's cell bank) is planted in 96 orifice plates uniformly, every porocyte number is 10000.
2) treat adherent, the rear dosing of spending the night, dosing (8-diethyl malonate berberine concentration is respectively 50,16.67,5.56,1.85,0.62 μ M), each concentration has 3 multiple holes.
3) cultivated 48 hours, complete medium is replaced to the mixture (10: 1) of serum-free medium and CCK8, hatched 2 hours in 37 ℃ of incubators.
4) take 450nm as measuring wavelength, take 650nm as the contrast wavelength, measure reading on microplate reader.
Result:
8-diethyl malonate berberine is 22.59 μ M to the IC50 value of K562 cell.
Claims (10)
1.8-the application of diethyl malonate berberine in the preparation antitumor drug.
2. application as claimed in claim 1, is characterized in that, described tumor is leukemia or cancer of pancreas.
3. application as claimed in claim 1, is characterized in that, described antitumor drug is anti-leukemia medicine.
4. application as claimed in claim 1, is characterized in that, described antitumor drug is anti-cancer of pancreas medicine.
5. application as claimed in claim 1, is characterized in that, the dosage form of described antitumor drug is injection, tablet or capsule.
6. a method that suppresses tumor cell in vitro propagation, is characterized in that, 8-diethyl malonate berberine is added in the culture fluid of tumor cell.
7. method as claimed in claim 6, is characterized in that, described tumor cell is leukemia or pancreatic cancer cell.
8. method as claimed in claim 6, is characterized in that, the final concentration that adds 8-diethyl malonate berberine is 1-50 μ M.
9. an antitumor drug, is characterized in that, the active component of described antitumor drug is 8-diethyl malonate berberine.
10. antitumor drug as claimed in claim 9, is characterized in that, described tumor is leukemia or cancer of pancreas.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011103750311A CN103127101A (en) | 2011-11-22 | 2011-11-22 | Application of 8-diethyl malonate berberine in preparation of antitumor drug |
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| CN2011103750311A CN103127101A (en) | 2011-11-22 | 2011-11-22 | Application of 8-diethyl malonate berberine in preparation of antitumor drug |
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| CN2011103750311A Pending CN103127101A (en) | 2011-11-22 | 2011-11-22 | Application of 8-diethyl malonate berberine in preparation of antitumor drug |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101870694A (en) * | 2009-04-21 | 2010-10-27 | 中国科学院上海药物研究所 | 8,8-disubstituted-13,13a-dihyrdroberberine derivative as well as pharmaceutical composition and application thereof |
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- 2011-11-22 CN CN2011103750311A patent/CN103127101A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101870694A (en) * | 2009-04-21 | 2010-10-27 | 中国科学院上海药物研究所 | 8,8-disubstituted-13,13a-dihyrdroberberine derivative as well as pharmaceutical composition and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 李波 等: ""小檗碱及其衍生物的研究进展", 《药学学报》, vol. 43, no. 8, 31 December 2008 (2008-12-31), pages 773 - 787 * |
| 杨勇等: "8-烷基小檗碱的合成", 《有机化学》, vol. 27, no. 11, 31 December 2007 (2007-12-31), pages 1438 - 1440 * |
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Application publication date: 20130605 |