CN102688493A - Pharmaceutical composition containing resveratrol, resveratrol derivative and Bc1-2 inhibitor, and application thereof - Google Patents
Pharmaceutical composition containing resveratrol, resveratrol derivative and Bc1-2 inhibitor, and application thereof Download PDFInfo
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- CN102688493A CN102688493A CN201110080495XA CN201110080495A CN102688493A CN 102688493 A CN102688493 A CN 102688493A CN 201110080495X A CN201110080495X A CN 201110080495XA CN 201110080495 A CN201110080495 A CN 201110080495A CN 102688493 A CN102688493 A CN 102688493A
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Abstract
The present invention relates to a pharmaceutical composition containing resveratrol, a resveratrol derivative and a Bc1-2 inhibitor, and applications of the pharmaceutical composition for preparing pharmaceuticals to cure colorectal carcinoma, liver cancer, lung cancer, kidney cancer, stomach cancer, brain tumor, sarcoma, neuroglioma, pancreas cancer, ovary cancer, breast cancer or prostate cancer. The pharmaceutical composition has an obvious synergistic effect, increases the pharmic curative effect, reduces the administration dosage and reduces the adverse reactions.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition and the application in the medicine of preparation treatment cancer thereof, be specifically related to contain the pharmaceutical composition and the application in the medicine of preparation treatment colon cancer, hepatocarcinoma, pulmonary carcinoma, renal carcinoma, gastric cancer, cerebroma, sarcoma, glioma, cancer of pancreas, ovarian cancer, breast carcinoma or carcinoma of prostate thereof of resveratrol and resveratrol analog derivative and Bcl-2 inhibitor.
Background technology
World Health Organization's investigation report shows that global cancer condition is serious day by day, and 20 years from now on new patients' number will be increased to 1,500 ten thousand by present every year 1000 ten thousand, because of the number that cancer is dead also will be by increasing to 1,000 ten thousand 6,000,000 of every year.Primary hepatocarcinoma is one of human modal malignant tumor for occurring in the epithelial canceration of hepatocyte and stones in intrahepatic bile duct; The morbidity of colon cancer and environment, living habit, especially diet style is relevant, it is generally acknowledged that high fat diet and cellulose deficiency are main pathogenic factors, along with growth in the living standard, the change of dietary structure, the sickness rate of colon cancer is ascendant trend year by year; Glioma is to originate from neurogliocyte, betides neuroectodermal tumor, and its principal character is the growth of tumor cell diffuse infiltrating, does not have clear and definite border, infinite multiplication and have the height aggressive.Although the neurosurgery skill is constantly perfect in recent years, radiotherapy is accurately located and the continuous research and development of chemotherapeutics, glioma patient more after still barely satisfactory, the medicine of therefore studying glioma is extremely urgent.
The antitumor drug that has gone on the market at present is more, and like alkylating agent medicine, antimetabolite, AGPM, immunomodulator etc., but medicine is because toxicity is bigger mostly, and patient does not tolerate.Lots of clinical facts have proved that malignant tumor can be effectively treated in the Chinese medicine or the combination of Chinese and Western medicine, can alleviate the toxic and side effects of chemicotherapy simultaneously.Utilization modern medicine means find that some bioactive natural products can effectively suppress the growth of tumor cell, have the effect of cell death inducing.Numerous antibiotic and the antitumor drug that uses at present or be directed to natural product, or get through its structure of modification.Therefore, safe bioactive natural product applies to clinically will have broad prospects with the treatment cancer.Along with the Study on Molecular Mechanism to the incidence and development of tumor is more and more clearer, the multiple malignant tumor of molecular targeted treatment has received to be paid close attention to widely and pays much attention to.The molecular targeted agents selectivity is high, wide spectrum is effective, and its safety is superior to the cytotoxicity chemotherapeutics, is the new direction of present oncotherapy field development.
Resveratrol (Resveratrol) is a kind of polyphenolic substance that extensively is present in the various plants such as Fructus Vitis viniferae, Rhizoma Polygoni Cuspidati, Semen arachidis hypogaeae, is a kind of natural phytoalexin, is the green cancer therapy drug of anti-curing oncoma.Experiment in vivo and vitro shows, resveratrol to most of tumors initial, breed, develop these three Main Stage and all have and suppress and even reverse effect.Its antitumor mechanism can play a role through antioxidation, retardance cell cycle, promotion apoptosis of tumor cells, inducing tumor cell differentiation, the activity that suppresses epoxide hydrolase and cytochrome enzyme P450, interference coherent signal transduction path, inhibition tumor-blood-vessel growth etc.
Apoptosis (program cell death) is the natural way that body is removed unusual or unwanted cells, if it is affected and then possibly causes the generation of various diseases such as cancer.The Bcl-2 family protein is the important regulator of apoptosis; Bcl-2 and Bcl-xL overexpression in polytype tumor wherein; Being considered to maybe be relevant with generation, development and the drug resistance generation of tumor, so become the research focus of antineoplaston in recent years to the drug development of Bcl-2 and Bcl-xL anti-apoptotic proteins.ABT-263 and ABT-737 are the micromolecule Bcl-2 inhibitor by U.S. Abbott Laboratories (Abbott) pharmacy exploitation, and be remarkable to the kinds of tumors effect, and the orally-ingestible use, has a good application prospect.
Along with the progress of oncomolecularbiology, the molecular targeted treatment of tumor has become the focus of tumor research, in the treatment of kinds of tumors, has brought into play important effect.Yet the biological behaviour of most of tumor is not to be arranged by single signal transduction pathway, but a plurality of signal transduction pathway concurs, and Chinese medicine receives publicity with the effect advantage of the many target spots of its polygenes just day by day.Therefore reasonable joint medication; The incomparable superiority of single medicament is arranged; Drug combination carries out targeted therapy to many target spots will not only be intended to minimizing or delay chemical sproof appearance, reduction toxicity, and through multiple medicine the synergism that cancerous cell kills and wounds obtained better therapeutic.
Summary of the invention
To above technological deficiency; The present invention provides a kind of pharmaceutical composition and the application in the medicine of preparation treatment cancer thereof, is specially the pharmaceutical composition and the application in the medicine of preparation treatment colon cancer, hepatocarcinoma, pulmonary carcinoma, renal carcinoma, gastric cancer, cerebroma, sarcoma, glioma, cancer of pancreas, ovarian cancer, breast carcinoma or carcinoma of prostate thereof that contain resveratrol and resveratrol analog derivative and Bcl-2 inhibitor.
The present invention contains in the pharmaceutical composition of resveratrol and resveratrol analog derivative and Bcl-2 inhibitor, and said resveratrol and resveratrol analog derivative are resveratrol; The Bcl-2 inhibitor can be ABT-263 or ABT-737, or both corresponding structure analog, derivant.
Resveratrol in the pharmaceutical composition of the present invention and resveratrol analog derivative are preferably resveratrol, and its corresponding structure formula is suc as formula shown in the I.
In the pharmaceutical composition of the present invention, said component is not limited to resveratrol medicine itself, can also be its pharmaceutically useful salt, hydrate or derivant etc.
Among the present invention, said Bcl-2 inhibitor can be preferably ABT-263 or ABT-737 for the medicine of the Bcl-2 inhibitor of any structure type.Wherein ABT-263 is the chemical compound shown in the formula II that is put down in writing among the US2007027135:
Wherein ABT-737 is the chemical compound shown in the formula III of being put down in writing among WO2005049594 and the WO2005049593:
In the pharmaceutical composition of the present invention, said component is not limited to above-mentioned ABT-263 and ABT-737 itself, can also be the salt of their hydrate, analog, derivant and other organic or inorganic.
The present invention contains in the pharmaceutical composition of resveratrol and resveratrol analog derivative and Bcl-2 inhibitor, and the mol ratio of resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 15.0-200.0: 0.5-4.0; The mol ratio of further preferred resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 30.0-100.0: 1.0-2.0.
The pharmaceutical composition that the present invention contains resveratrol and resveratrol analog derivative and Bc l-2 inhibitor can be used to treat various tumors, and said tumor includes but not limited to colon cancer, hepatocarcinoma, pulmonary carcinoma, renal carcinoma, gastric cancer, cerebroma, sarcoma, glioma, cancer of pancreas, ovarian cancer, breast carcinoma or carcinoma of prostate.
The pharmaceutical composition of preferred resveratrol of the present invention and resveratrol analog derivative and Bcl-2 inhibitor is used for preparing the application of treating colon cancer, hepatocarcinoma and gliomatous medicine.
In the application of pharmaceutical composition of the present invention in preparation treatment colon cancer, hepatocarcinoma and gliomatous medicine, the mol ratio of resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 30.0-100.0: 1.0-2.0; The mol ratio of preferred resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 50.0-100.0: 1.5-2.0; The mol ratio of further preferred resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 100.0: 2.0.
Contain resveratrol and resveratrol analog derivative and Bcl-2 inhibitor combination in the application of the medicine of preparation treatment colon cancer, hepatocarcinoma, pulmonary carcinoma, renal carcinoma, gastric cancer, cerebroma, sarcoma, glioma, cancer of pancreas, ovarian cancer, breast carcinoma or carcinoma of prostate; In the scheme of the medicament of the present composition being processed administration simultaneously; Resveratrol and resveratrol analog derivative and Bcl-2 inhibitor can be contained in in a kind of pharmaceutical preparation such as tablet or the capsule; Also can resveratrol and resveratrol analog derivative and Bcl-2 inhibitor be made preparation respectively; As make tablet or capsule respectively; And adopting the conventional mode in this area with their packings or combine, the patient takes according to the indication of package insert then simultaneously; In the scheme of the medicament of the present composition being processed administration successively; Can resveratrol and resveratrol analog derivative be made different preparations respectively with the Bcl-2 inhibitor; And adopt the conventional mode in this area with their packings or combine; The patient takes according to the sequencing of package insert indication then; Or two kinds of compositions in the above-mentioned composition are processed a kind of preparation of controlled release, and a kind of composition in the first release composition and then the another kind of composition in the release composition, the patient only need take this controlled release composition preparation; In the scheme of the medicament that the present composition is prepared into the intersection administration; Can resveratrol and resveratrol analog derivative be made different preparations respectively with the Bcl-2 inhibitor; And adopt the conventional mode in this area with their packings or combine; The patient takes according to the chi sequence of package insert indication then, the controlled release preparation that perhaps becomes resveratrol and resveratrol analog derivative and Bcl-2 inhibitor intersection to discharge this preparation of pharmaceutical compositions.
In the application in the medicine of preparation treatment colon cancer, hepatocarcinoma, pulmonary carcinoma, renal carcinoma, gastric cancer, cerebroma, sarcoma, glioma, cancer of pancreas, ovarian cancer, breast carcinoma or carcinoma of prostate of resveratrol and resveratrol analog derivative and Bcl-2 inhibitor combination; Resveratrol in the said compositions and resveratrol analog derivative and Bcl-2 inhibitor can use or with the using in order of any priority simultaneously, as can resveratrol and resveratrol analog derivative and Bcl-2 inhibitor being taken to the patient simultaneously; Also can earlier the Bcl-2 inhibitor taken, taken then to resveratrol and resveratrol analog derivative medicine to the patient; Or take the Bcl-2 inhibitor earlier, take resveratrol and resveratrol analog derivative medicine then; The interval of taking for both does not have special demands, but the interval of preferably taking two kinds of medicines is no more than one day; Perhaps two kinds of medicines replace administration.
Among the present invention; Can resveratrol of the present invention and the conventional method of resveratrol analog derivative and Bcl-2 inhibitor employing this area be prepared into the pharmaceutical preparation that is suitable for gastrointestinal administration or parenteral administration; The pharmaceutical preparation that the present invention preferably processes gastrointestinal administration with resveratrol and resveratrol analog derivative and Bcl-2 inhibitor, its dosage form can be conventional tablet or capsule or controlled release, slow releasing preparation.In the pharmaceutical preparation of resveratrol of the present invention and resveratrol analog derivative and Bcl-2 inhibitor combination, according to different dosage forms and preparation specification, the content of said compositions in preparation can be counted 1-99% for quality, is preferably 10%-90%; The adjuvant that preparation uses can adopt the conventional adjuvant in this area, reacts or the curative effect that do not influence medicine of the present invention is a prerequisite with the discord present composition; The method for preparing of said preparation can adopt the conventional method for preparing in this area to prepare.
Among the present invention; The not special restriction of preparation of compositions method; Resveratrol and resveratrol analog derivative and Bcl-2 inhibitor can directly mix makes preparation then; Or respectively and/or corresponding auxiliary material mix and to make preparation respectively, and then packaging together, or mix and then mix and make preparation with corresponding auxiliary material respectively according to the conventional mode in this area.
The dosage of the pharmaceutical composition among the present invention can carry out suitable variation according to the dosage form difference of administration object, route of administration or medicine, but is prerequisite to guarantee that this pharmaceutical composition can reach effective blood drug level in mammalian body.
The present invention has carried out the test that resveratrol and resveratrol analog derivative and the combination of Bcl-2 inhibitor kill DLD1 (colon cancer cell line), HUH-7 (hepatoma cell strain) and U251 (neuroglial cytoma strain) respectively; The result shows; Resveratrol of the present invention and resveratrol analog derivative and Bcl-2 inhibitor combined therapy colon cancer, hepatocarcinoma and glioma have significant cooperative effect; Improve the curative effect of medicine, reduced dosage, reduced the generation of side effect.
The specific embodiment
In conjunction with following examples the present invention is done further elaboration, but the present invention is not limited to this.
Embodiment
Reagent and method:
Cell: DLD1 (colon cancer cell line), HUH-7 (hepatoma cell strain) and U251 (neuroglial cytoma strain) are all available from American Type Culture Collection (ATCC), Maryland, USA Rockwell.
Medicine: institute's pharmaceutical composition is all by following method 1 or 2 said preparations of method in following examples; Resveratrol replaces this Chinese mugwort institute of Chinese materia medica available from Nanjing; The Bcl-2 inhibitor all is synthesized into by document, and ABT-263 and ABT-737 synthesized reference document are: Synthesis, 15,2398-2404, WO2005049594, WO2005049593 and US2007027135.
Method 1: each component of accurate weighing corresponding pharmaceutical compositions, dissolve respectively with dimethyl sulfoxide, be made into the stock solution of 10mM separately; Preserve down at-20 ℃; Be diluted to suitable concentration with fresh culture medium during use, the solution of each component of 1 microlitre of respectively asking for then mixes subsequent use.In all tests, the ultimate density of dimethyl sulfoxide is answered≤5g/L, so that do not influence cell activity.
With all cells in RPMI 1640 culture medium that contain 10% calf serum, 100kU/L penicillin, 100mg/L streptomycin, 37 ℃, 5%CO
2Damp condition cultivate down, in the previous day of dosing, on six orifice plates, carry out cell inoculation 2 * 10
5/ hole adds the pharmaceutical composition solution of preparation as stated above then in cell, make each component reach its working concentration, specifically sees 1-6 in the table 1.
After the drug treating, measure cell death through trypan blue (Trypan Blue), cell turns into 10 minutes through carrying out pancreatin at 37 ℃ with trypsin sodium/EDTA.Get into the culture medium because dead cell comes off from incubator,, and then, mix with the trypan blue dyestuff with culture medium suspended sediment again through all cells of centrifugal collection under 1200 rev/mins.After the dyeing, count with optical microscope and hematimeter.Dyed the blue dead cell of counting by dyestuff.500 cells of picked at random are counted, and dead cell is recently expressed with the percentage that accounts for the grand total cell.
Method 2: each component of accurate weighing corresponding pharmaceutical compositions, dissolve respectively with dimethyl sulfoxide, be made into the stock solution of 10mM separately, preserve down at-20 ℃.Be diluted to suitable concentration with fresh culture medium during use, the solution for standby of each component of 1 microlitre of respectively asking for then.In all tests, the ultimate density of dimethyl sulfoxide is answered≤5g/L, so that do not influence cell activity.
With all cells in RPMI 1640 culture medium that contain 10% calf serum, 100kU/L penicillin, 100mg/L streptomycin, 37 ℃, 5%CO
2Damp condition cultivate down, in the previous day of dosing, on six orifice plates, carry out cell inoculation 2 * 10
5/ hole adds each component solution of the pharmaceutical composition of preparation as stated above with any order then in cell, make each component reach its working concentration, specifically sees 7-12 in the table 1.
After the drug treating, measure cell death through trypan blue (Trypan Blue), cell turns into 10 minutes through carrying out pancreatin at 37 ℃ with trypsin sodium/EDTA.Get into the culture medium because dead cell comes off from incubator,, and then, mix with the trypan blue dyestuff with culture medium suspended sediment again through all cells of centrifugal collection under 1200 rev/mins.After the dyeing, count with optical microscope and hematimeter.Dyed the blue dead cell of counting by dyestuff.500 cells of picked at random are counted, and dead cell is recently expressed with the percentage that accounts for the grand total cell.
Tabulate down in the drug regimen shown in 1, the combination of 1-6 is by method 1 preparation, and the combination of 7-12 is by method 2 preparations.
Table 1
The resveratrol of embodiment 1 different proportion and the combination Synergistic of ABT-263 promote the test of DLD1 cell death, see table 2.
Table 2
Cause in the test of colon cancer cell line DLD1 cell death at the investigation related compound; Find almost acellular death when use 2.0 μ M ABT-263 or lower concentration separately, even only have an appointment 20% cell death when using 100.0 μ M resveratrols separately; (50.0 μ M resveratrols+1.5 μ M ABT-263) then produce the obvious synergistic effect when both share under low concentration, cause 48% cancer cell death; When both share with the ratio of 100.0 μ M resveratrols+2.0 μ M ABT-263, then produce significant more synergism, cause 68% cancer cell death.
The resveratrol of embodiment 2 different proportions and the combination Synergistic of ABT-737 promote the test of DLD1 cell death, see table 3.
Table 3
Cause in the test of colon cancer cell line DLD1 cell death the cell death of the 15-20% that finds when use 2.0 μ M ABT-737 or 100.0 μ M resveratrols separately, only to have an appointment investigating related compound; (50.0 μ M resveratrols+1.5 μ M ABT-737) then produce than the obvious synergistic effect when both share under low concentration, cause 31% cancer cell death; When both share with the ratio of 100.0 μ M resveratrols+2.0 μ M ABT-737, then produce significant more synergism, cause 55% cancer cell death.
The resveratrol of embodiment 3 different proportions and the combination Synergistic of ABT-263 promote the test of HUH-7 cell death, see table 4.
Table 4
Cause in the test of hepatoma cell strain HUH-7 cell death at the investigation related compound; 10% the cell death of finding when use 2.0 μ M ABT-263 or lower concentration separately, only to have an appointment; Almost there is not cell death when separately using 50.0 μ M resveratrols, even only have an appointment 25% cell death when using 100.0 μ M resveratrols separately; (50.0 μ M resveratrols+1.5 μ M ABT-263) then produce than the obvious synergistic effect when both share under low concentration, cause 31% cancer cell death; When both share with the ratio of 100.0 μ M resveratrols+2.0 μ M ABT-263, then produce significant more synergism, cause 86% cancer cell death.
The resveratrol of embodiment 4 different proportions and the combination Synergistic of ABT-263 promote the test of U251 cell death, see table 5.
Table 5
Cause in the test of neuroglial cytoma strain U251 cell death 10% the cell death of finding when use 1.5 μ M ABT-263 or 50.0 μ M resveratrols separately, only to have an appointment investigating related compound; The cell death of about 20-35% when using 2.0 μ M ABT-263 or 100.0 μ M resveratrols separately; (50.0 μ M resveratrols+1.5 μ M ABT-263) then produce the obvious synergistic effect when both share under low concentration, cause 44% cancer cell death; When both share with the ratio of 100.0 μ M resveratrols+2.0 μ M ABT-263, then produce significant more synergism, cause 90% cancer cell death.
Although the foregoing description describes in detail technical scheme of the present invention; But technical scheme of the present invention is not limited to above embodiment; Under the situation that does not break away from thought of the present invention and aim, any change that technical scheme of the present invention is done all will fall into claims of the present invention institute restricted portion.
Claims (10)
1. a pharmaceutical composition is characterized in that, said pharmaceutical composition contains resveratrol and resveratrol analog derivative and Bcl-2 inhibitor.
2. pharmaceutical composition according to claim 1 is characterized in that, the mol ratio of said resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 15.0-200.0: 0.5-4.0.
3. pharmaceutical composition according to claim 2 is characterized in that, the mol ratio of said resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 30.0-100.0: 1.0-2.0.
4. according to each described pharmaceutical composition of claim 1-3, it is characterized in that said resveratrol and resveratrol analog derivative are resveratrol; Said Bcl-2 inhibitor is ABT-263 or ABT-737.
5. the application of each described pharmaceutical composition of claim 1-4 in the medicine of preparation treatment cancer.
6. application according to claim 5 is characterized in that, said cancer is colon cancer, hepatocarcinoma, pulmonary carcinoma, renal carcinoma, gastric cancer, cerebroma, sarcoma, glioma, cancer of pancreas, ovarian cancer, breast carcinoma or carcinoma of prostate.
7. application according to claim 6; It is characterized in that; In the application in preparation treatment colon cancer, hepatocarcinoma and gliomatous medicine, the mol ratio of said resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 30.0-100.0: 1.0-2.0.
8. application according to claim 7 is characterized in that, the mol ratio of said resveratrol and resveratrol analog derivative and Bcl-2 inhibitor is 50.0-100.0: 1.5-2.0.
9. application according to claim 8 is characterized in that, said resveratrol and resveratrol analog derivative are resveratrol; Said Bcl-2 inhibitor is ABT-263 or ABT-737.
10. according to each described application of claim 5-9, it is characterized in that resveratrol in the said pharmaceutical composition and resveratrol analog derivative and Bcl-2 inhibitor use or using in order with any priority simultaneously.
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Cited By (5)
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| CN104288156A (en) * | 2014-09-22 | 2015-01-21 | 新乡医学院 | Pharmaceutical composition for treating stomach cancer and application thereof |
| CN106967049A (en) * | 2017-03-22 | 2017-07-21 | 杭州师范大学 | A kind of resveratrol line fluorescent mark molecule and its synthetic method |
| US10105357B2 (en) * | 2014-10-24 | 2018-10-23 | Launx Biomedical Co., Ltd. | Indication of antibiotic drugs for preparation of cancer inhibition pharmaceutical composition |
| WO2019108773A1 (en) * | 2017-11-29 | 2019-06-06 | Stc.Unm | Substituted stilbenes as inhibitors of nf-kb and activators of nrf2 |
| CN112789036A (en) * | 2018-08-02 | 2021-05-11 | G·拉瓦尼安 | Compositions comprising resveratrol glycosides and curcuminoids |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104288156A (en) * | 2014-09-22 | 2015-01-21 | 新乡医学院 | Pharmaceutical composition for treating stomach cancer and application thereof |
| CN104288156B (en) * | 2014-09-22 | 2017-01-18 | 新乡医学院 | Pharmaceutical composition for treating stomach cancer and application thereof |
| US10105357B2 (en) * | 2014-10-24 | 2018-10-23 | Launx Biomedical Co., Ltd. | Indication of antibiotic drugs for preparation of cancer inhibition pharmaceutical composition |
| CN106967049A (en) * | 2017-03-22 | 2017-07-21 | 杭州师范大学 | A kind of resveratrol line fluorescent mark molecule and its synthetic method |
| CN106967049B (en) * | 2017-03-22 | 2018-06-26 | 杭州师范大学 | A kind of resveratrol line fluorescent mark molecule and its synthetic method |
| US10125119B2 (en) | 2017-03-22 | 2018-11-13 | Hangzhou Normal University | Series of resveratrol-derivative fluorescently labeled molecules and synthesis method thereof |
| WO2019108773A1 (en) * | 2017-11-29 | 2019-06-06 | Stc.Unm | Substituted stilbenes as inhibitors of nf-kb and activators of nrf2 |
| CN112789036A (en) * | 2018-08-02 | 2021-05-11 | G·拉瓦尼安 | Compositions comprising resveratrol glycosides and curcuminoids |
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