CN106902101A - Application of the thelephora ganbajun in antineoplastic sensitizer is prepared - Google Patents
Application of the thelephora ganbajun in antineoplastic sensitizer is prepared Download PDFInfo
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- CN106902101A CN106902101A CN201510975676.7A CN201510975676A CN106902101A CN 106902101 A CN106902101 A CN 106902101A CN 201510975676 A CN201510975676 A CN 201510975676A CN 106902101 A CN106902101 A CN 106902101A
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- antineoplastic
- thelephora ganbajun
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/12—Ketones
- A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/475—Quinolines; Isoquinolines having an indole ring, e.g. yohimbine, reserpine, strychnine, vinblastine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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Abstract
The invention belongs to cell biology and field of medicaments, it is related to application of the thelephora ganbajun in antineoplastic sensitizer is prepared.Thelephora ganbajun is natural products, the effect with reversing multiple medicine resistance of tumor cells, can be as the reversal agent of tumor multi-medicine drug-resistant；Thelephora ganbajun also has increases the effect of tumor multi-medicine drug-resistant cells against neoplastic drug susceptibility, can be used as chemotherapeutic sensitizer.Present invention also offers the method that the pharmaceutical composition that antineoplastic and thelephora ganbajun are used in combination suppresses tumor multi-medicine drug-resistant cell propagation.Micromolecular compound thelephora ganbajun in the present invention is developed as new antineoplastic or its auxiliary element, and substantially, environmental protection will provide a kind of new approach and means to tumor killing effect to treat and curing tumour.
Description
Technical field
The invention belongs to cell biology and field of medicaments field, specifically, the present invention relates to the new use of thelephora ganbajun
On the way.The invention further relates to corresponding pharmaceutical composition and its application process.
Background technology
Tumour is body under the effect of various carcinogenic factors, and some cell of local organization loses right on gene level
The normal regulation of its growth, the abnormality for causing its clonal abnormality hyperplasia and being formed.The neoplastic disease number of cases of China is quite huge
Greatly, there is data to show and account for the 55% of whole world case load.
Influence of the benign tumour to body is smaller, is mainly shown as local compression and obstructive symptom.Malignant tumour is due to dividing
Change immature, growth very fast, infiltration destroys the 26S Proteasome Structure and Function of organ, and can shift, thus serious on body influence.Dislike
Property tumour can have heating, intractable pain in addition to it can cause local compression and the obstructive symptom similar to above-mentioned benign tumour, also,
Late period may occur in which seriously become thin, the state of weak, anaemia and cachexia.
The traditional Chinese medical science thinks that the cause of cancer is equilibrium between yin and yang in human body first, and histocyte is long-term in different carcinogenic factors
Under effect, cell mutation and cause.Cancerous tissue is also a part for human body in fact, only in this imbalance between YIN and YANG of people, five
Raw gram of row multiplies on the premise of insult changes, and the immunosurveillance system of human body can just lose monitoring to it, develop as one pleases.Chinese herbal medicine
Can with recuperating qi-blood, adjustment equilibrium between yin and yang, maintain normal vital signs and save one's life；To train correction gas, produce antibody, cleaning "
Malicious source " and effect a permanent cure.Thus, Chinese herbal medicine extract turns into treatment one important directions of tumour.
Thelephora ganbajun is natural products, and bioavilability is high, Nature comparison is stable, with Clinical practice value.With people
Going deep into the research of its chemistry and biology, its molecular mechanism of action will progressively clearly, and this will further promote such
The modifying for chemical structure and structure activity study of compound, and it is favorably improved its medical value.
The content of the invention
It is an object of the invention to provide the new medicinal usage of thelephora ganbajun.
It is an object of the invention to provide a kind of antineoplastic sensitizer.
On the one hand, the application the invention provides thelephora ganbajun in antineoplastic sensitizer is prepared.Wherein, wizened bacterium
The structure of element is as shown in Figure 1.
Described tumour includes carcinoma of mouth, breast cancer, liver cancer, lung cancer, cervical carcinoma, colon cancer or cancer of pancreas.
Described antineoplastic is vincristine, daunorubicin, taxol or 5 FU 5 fluorouracil.
Thelephora ganbajun can be as tumor multi-drug resistance reversal agents.
On the other hand, the invention provides a kind of antineoplastic pharmaceutical compositions, described antineoplastic pharmaceutical compositions have
Effect composition is antineoplastic and thelephora ganbajun.
Described antineoplastic is CCSA or cell cycle nonspecific agent (CCNSA).
Described antineoplastic is vincristine, daunorubicin, taxol or 5 FU 5 fluorouracil.
Present invention also offers a kind of method for suppressing tumor cell in vitro growing multiplication, i.e., in the culture medium of tumour cell
It is middle to add thelephora ganbajun and antineoplastic.
Wherein, the final concentration of 1-100 μm of ol/L of thelephora ganbajun is added.
Described tumour includes carcinoma of mouth, breast cancer, liver cancer, lung cancer, cervical carcinoma or cancer of pancreas.
When implementing the above method, thelephora ganbajun is first added, be subsequently adding antineoplastic.Wizened bacterium can also simultaneously be added
Element and antineoplastic.
The invention provides the new application of thelephora ganbajun, i.e. its application in antineoplastic sensitizer is prepared.It is wizened
Rhzomorph is natural products, the effect with reversing multiple medicine resistance of tumor cells, can be as the reversal agent of tumor multi-medicine drug-resistant；It is dry
Bar rhzomorph also has increases the effect of tumor multi-medicine drug-resistant cells against neoplastic drug susceptibility, can make as chemotherapeutic sensitizer
With.Micromolecular compound thelephora ganbajun in the present invention is developed as new antineoplastic or its auxiliary element, suppression
Substantially, environmental protection will provide a kind of new approach and means to knurl effect to treat and curing tumour.
Brief description of the drawings
Fig. 1 is the structure of thelephora ganbajun.
Specific embodiment
The invention provides a kind of antineoplastic, the active component of described antineoplastic is thelephora ganbajun.It is described
Tumour can be HCC, stomach cancer cell, cervical cancer cell or blood cell.
Micromolecular compound of the invention can be prepared using various conventional preparation methods.For example, using artificial chemistry
The method of synthesis.
Using micromolecular compound of the present invention, by various conventional screening assays, can filter out and phase occurs with thelephora ganbajun
The material of interaction, such as acceptor, inhibitor or antagonist.
The present invention and its inhibitor, antagonist etc., when (administration) is administered in treatment, it is possible to provide different effects.
Generally, these materials can be formulated in nontoxic, inert and pharmaceutically acceptable aqueous carrier medium, wherein pH is usual
About 5-8, preferably pH be about 6-8, pH value can be varied from the property for being formulated material and illness to be treated.
The pharmaceutical composition for preparing can be administered by conventional route, including（But it is not limited to）：Intramuscular, intraperitoneal,
Subcutaneous, intracutaneous or local administration.
By taking thelephora ganbajun of the invention as an example, it can be combined with suitable pharmaceutically acceptable carrier.This kind of medicine
Compositions contain the compound and pharmaceutically acceptable carrier or excipient of therapeutically effective amount.This kind of carrier includes（But simultaneously
It is not limited to）：Salt solution, buffer solution, glucose, water, glycerine, ethanol, and combinations thereof.Pharmaceutical preparation should match with administering mode.
Thelephora ganbajun of the invention can be made into injection form, such as water-soluble with other assistant agents with physiological saline or containing glucose
Liquid is prepared by conventional method.The pharmaceutical composition of such as tablet and capsule etc, can be prepared by conventional method.
Pharmaceutical composition such as injection, solution, tablet and capsule are preferably aseptically manufactured.The dosage of active component is effective treatment
Amount, such as daily mg/kg body weight of about 1 microgram/kg body weight-about 5.Additionally, thelephora ganbajun of the invention can also be with other
Therapeutic agent is used together.Certainly, specific dosage is also contemplated that the factors such as method of administration, patient health situation, and these are all skilled
Within the scope of doctor's technical ability.
Experimental technique：
1. cell recovery
1) cryopreservation tube is taken out from liquid nitrogen container, in direct plungeing into 37 DEG C of warm water, and shake makes it melt as early as possible frequently.
2) take out cryopreservation tube from 37 DEG C of water-baths, with suction pipe suction out cell suspension, injection centrifuge tube and addition more than 10 times
Nutrient solution, low-speed centrifugal after mixing, abandons supernatant, repeats and is washed once with nutrient solution.
3) after suitably being diluted with nutrient solution, inoculated and cultured bottle is placed on 37 DEG C of incubator quiescent cultures, and next day changes culture
Liquid, continues to cultivate.Passed on when culture is to finite concentration.PANC-1 cell culture is containing 10% Gibico hyclones
In DMEM high glucose mediums；The cell culture such as K562, HGC, QGY, MCF-7, PC-3 are in 1640 cultures containing 10% hyclone
In base, SK-hep1, HeLa, HepG2, etc. cell culture in the DMEM high glucose mediums containing 10% hyclone, in culture medium
Penicillin containing 100U/ml and 100 μ g/ml streptomysins.
2. passage culture
The situation of daily observation of cell growth, passes on when when cell, length is converged to about 90% in blake bottle, is about passed every 2-4 days
In generation, is once.One bottle passes on into three bottles, or 25 cm2Pass in 75 cm2Blake bottle in.Method：
1) with 1 × phosphate buffer washed cell once.
2) digestion of 2-3ml pancreatin digestive juice is added, several minutes in 37 DEG C of incubators are placed in.Tissue Culture Flask is patted with hand,
Make cell separation.
3) terminate pancreatin with the suitable culture medium of the Gibico hyclones containing 10-15% to digest.Cell is sub-packed in newly
Blake bottle in, continue cultivate.
When suspension cell is passed on, centrifugation in centrifuge tube is collected directly from, abandons old culture medium.General one bottle passes on into three bottles
Ratio is sub-packed in new blake bottle, adds fresh culture to continue to cultivate.
3. cell cryopreservation
1) the cell Trypsin Induced of culture to exponential phase is taken, is collected in centrifuge tube and is counted, be centrifuged.
2) reject trypsase and old nutrient solution, what addition had been configured freezes nutrient solution (containing 10% DMSO, 40%
DMEM and 50% Gibico hyclones), the ultimate density of cell is 0.5- 1 × 10 in frozen stock solution7/ml.Gently blown with suction pipe
Beating makes cell uniform, is then distributed into aseptic cryopreservation tube, often pipe -1.5ml of Jia 1.
3) cryopreservation tube is put into freezing storing box and puts -80 DEG C of quick-frozen, preservations in immigration liquid nitrogen container after 5 hours.
4. experiment material
Medicine prepares：
Thelephora ganbajun is dissolved in DMSO (dimethyl sulfoxide (DMSO)), and the mother liquor for being configured to 100mM or 50mM is standby.
Thelephora ganbajun can be bought from Shanghai Yuan Mu bio tech ltd, and its structure is as shown in Figure 1.
Verapamil（VPL）, vincristine (VCR) and adriamycin (ADR) be purchased from Roche chemical company, purity is both greater than
99%。
Cell derived:
Human oral cancer cell line KB and its drug-resistant cell strain KB/VCR are provided by Chinese Academy of Sciences's medicine, Breast cancer lines MCF-
7 and its drug-resistant cell strain MCF-7/ADR, human colon carcinoma HCT-8 and its drug-resistant cell strain HCT-8/VCR are purchased from Nanjing Kai Jisheng
Thing company.
Kit:
CCK-8 kits are purchased from colleague company.
CCK-8 is tested
KB and KB/VCR cells and MCF-7 and MCF-7/ADR cells are all inoculated into 96 orifice plates according to the density in 3500/ hole,
The ADR of various concentrations after 24 h, VCR, thelephora ganbajun and VCR and thelephora ganbajun one are reinstated the α-MEM containing 10% hyclone and are matched somebody with somebody
It is added in each hole after good.After cultivating 48 h, nutrient solution is discarded, culture medium and 10 μs of the 90 μ L without serum are added per hole
L CCK-8 reagents.After 37 DEG C of 2 h of reaction, ELIASA reads the light absorption value of 450 nm wavelength（OD450）.By being calculated use
Cell growth fraction of the medicine group relative to control group.Blank group only adds culture medium to be not added with cell, and control group is addition and medicine
The DMSO of same volume, cell survival rate=（Experimental group OD450- blank groups OD450）/（Control group OD450- blank groups OD450）.
By IC50Value calculates resistance multiple again（Resistance Fold, RF）.
The IC of RF=drug-resistant cell strain50The IC of value/parental cell strain50Value.Each concentration sets 3 repeating holes, real
Test repetition 3 times.
All mdr cells grow 3 days before tying up to Cell suppression test in without pharmaceutical culture medium.Each numerical value is 3
Independent experiment result, IC50Represented in " means standard deviation " form.VCR, vincristine；ADR, daunorubicin；RF, resistance times
Number.
Embodiment 1
Experimental result：
KB/VCR and MCF-7/ADR are two kinds of conventional multidrug resistance cell strains, and in the present embodiment, both cells are also showed
Go out the characteristic of MDR.As shown in table 1, KB/VCR cells are respectively relative to KB cells to the resistance multiple of VCR and ADR
81.9 times and 94.4 times, and MCF-7/ADR cells are respectively 38.1 times compared to the resistance multiple to VCR and ADR with MCF-7 cells
With 20.9 times, display experiment mdr cell used has multidrug resistance, and MDR activity similar to the result of document report.
Half-inhibition concentration IC of the thelephora ganbajun to parental cell strain KB and MCF-750Respectively 169.88 μ Μ and 170.28
μ Μ, to the IC of drug-resistant cell strain KB/VCR and MCF-7/ADR50Respectively 191.15 μ Μ and 185.40 μ Μ, between the two without
Difference, multidrug resistance cell strain KB/VCR and MCF-7/ADR do not show resistance to the intersection of compound thelephora ganbajun
Medicine, illustrates that thelephora ganbajun can escape the multidrug resistance of mdr cell.
Conclusion：Thelephora ganbajun can overcome the drug resistance of drug-resistant cell strain, sensitiveness of the drug-resistant cell strain to thelephora ganbajun
It is essentially identical.
Embodiment 2：CCK-8 methods detect reverse effect of the thelephora ganbajun to tumor cell multidrug resistance activity
KB/VCR, MCF-7/ADR and HCT-8/VCR cell are inoculated into 96 orifice plates according to the density in 3500/ hole, after 24 h not
Reinstated after the α-MEM containing 10% hyclone are prepared with the VCR and thelephora ganbajun one of the VCR of concentration, and various concentrations and be added to
In each hole.After cultivating 48 h, discard nutrient solution, with method in embodiment 1, with CCK-8 kits survey drug-resistant cell strain and
Its parental cell plots curve to the activity of VCR and VCR+ thelephora ganbajuns.Each concentration sets 3 repeating holes, and experiment repeats 3
It is secondary.
Experimental result：
KB/VCR cells are 81.9 times to the resistance multiple of VCR, and MCF-7/ADR is 20.9 times to the resistance multiple of ADR.Work as addition
After 20 μM of thelephora ganbajun, make sensitiveness increase of the KB/VCR cells to VCR not notable, but ADR is to mdr cell MCF-7/
The sensitiveness of ADR increased 4.16 times.Thelephora ganbajun can reverse multiple drug resistance of tumor cell to the drug resistance of chemotherapeutics.
Conclusion：
Thelephora ganbajun can be with the drug resistance of reverse multiple drug resistance of tumor cell MCF-7/ADR, can be as the increasing of antineoplastic
Quick dose, or it is made pharmaceutical composition with antineoplastic.
The all documents referred in the present invention are all incorporated as reference in this application, independent just as each document
It is incorporated as with reference to such.In addition, it is to be understood that after above-mentioned instruction content of the invention has been read, those skilled in the art can
Made various changes or modifications with to the present invention, these equivalent form of values equally fall within the model that the application appended claims are limited
Enclose.
Claims (10)
1. application of the thelephora ganbajun in antineoplastic is prepared, described application is that thelephora ganbajun is preparing antineoplastic increasing
Application in quick dose.
2. application as claimed in claim 1, it is characterised in that thelephora ganbajun is tumor multi-drug resistance reversal agents.
3. application as claimed in claim 1, it is characterised in that described tumour includes carcinoma of mouth, nasopharyngeal carcinoma, breast cancer, liver
Cancer, lung cancer, cervical carcinoma, colon cancer or cancer of pancreas.
4. application as claimed in claim 1, it is characterised in that described antineoplastic is vincristine, daunorubicin, purple
China fir alcohol or 5 FU 5 fluorouracil.
5. a kind of antineoplastic pharmaceutical compositions, it is characterised in that the active ingredient of described antineoplastic pharmaceutical compositions is anti-swollen
Tumor medicine and thelephora ganbajun.
6. antineoplastic pharmaceutical compositions as claimed in claim 5, it is characterised in that described antineoplastic is the cell cycle
Specific drug or cell cycle nonspecific agent (CCNSA).
7. antineoplastic pharmaceutical compositions as claimed in claim 5, it is characterised in that described antineoplastic is new Changchun
Alkali, daunorubicin, taxol or 5 FU 5 fluorouracil.
8. it is a kind of suppress tumor cell in vitro growing multiplication method, it is characterised in that in the culture medium of tumour cell add
Thelephora ganbajun and antineoplastic.
9. method as claimed in claim 8, it is characterised in that described tumour includes carcinoma of mouth, breast cancer, nasopharyngeal carcinoma, liver
Cancer, lung cancer, cervical carcinoma or cancer of pancreas.
10. method as claimed in claim 8, it is characterised in that first add thelephora ganbajun, be subsequently adding antineoplastic.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510975676.7A CN106902101A (en) | 2015-12-23 | 2015-12-23 | Application of the thelephora ganbajun in antineoplastic sensitizer is prepared |
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| CN201510975676.7A CN106902101A (en) | 2015-12-23 | 2015-12-23 | Application of the thelephora ganbajun in antineoplastic sensitizer is prepared |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111315446A (en) * | 2017-08-08 | 2020-06-19 | 中山大学 | Methods and compositions for treating multidrug resistant tumors |
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2015
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111315446A (en) * | 2017-08-08 | 2020-06-19 | 中山大学 | Methods and compositions for treating multidrug resistant tumors |
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