CN104721210B - Application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared - Google Patents
Application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared Download PDFInfo
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- CN104721210B CN104721210B CN201510114766.7A CN201510114766A CN104721210B CN 104721210 B CN104721210 B CN 104721210B CN 201510114766 A CN201510114766 A CN 201510114766A CN 104721210 B CN104721210 B CN 104721210B
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Abstract
The invention belongs to antineoplastic field, application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared is disclosed.Application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared is proposed according to the present invention, oroxylin glycosides can be prepared into the medicine of the anti-tumor metastasis of various formulations, more preferable selection is provided for tumour patient.
Description
Technical field
The invention belongs to antineoplastic field, it is related to application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared.
Background technology
Metastases are multistage complicated processes, including the coming off of tumour cell, migrate, stick, grow
All influenceed and regulation and control by different factors in, each stage, be related to various adhesion molecules, stromatin enzyme, cell factor
And corresponding signal transduction mechanism and related gene alteration.Such as tumour cell relies on multiple protein enzyme early stage transfer,
Especially matrix metalloproteinase and urokinase degradation of cell epimatrix and basilar memebrane.Then, tumour cell is by different adhesions
Molecular action produce external force and itself some motion associated proteins and depart from original tissue, intrusion blood vessel or lymphatic vessel in
And distally organ and tissue are swum, now adhesion molecule assists the attached wall of tumour cell and finally swims out of tube wall, so as to form one
Transfer stove.But, not all antineoplastic is all acted on by anti-tumor metastasis mechanisms play, and some medicines are to original
Morbidity kitchen range have preferable therapeutic effect, but without the effect for suppressing tumour DISTANT METASTASES IN, some medicines, which have, preferably suppresses swollen
Tumor metastasis is acted on, but not good to the therapeutic action of primary lesion tumour, such as simply influences coming off, migrate, gluing for tumour cell
The a certain factor in the stage such as attached, without influenceing the growth of tumour cell, therefore can suppress metastases but not necessarily suppress tumour
Growth.In addition, the transfer of tumour has certain relation with the biological characteristics of tumour cell in itself, some tumours such as incidence
Tumour rarely has transfer based on local infiltration and recurrence, and easily occurs to turn if some tumours such as ED-SCLC, osteosarcoma etc.
Move.In addition, the type of tumour is different, the approach of transfer is also different, and in general, cancer is common with lymphatic metastasis, and sarcoma is then
Using Blood route metastasis to be common, some tumours are then by Implantation matastasis form, and these difference are produced to the treatment and prognosis of tumour
Different influences, therefore, reasonable selection relative medicine are the keys of tumour successful treatment, can specify medicine belong to which kind of resist
Tumour medicine helps to carry out the rational use of medicines for different types of tumour.
Oroxylin is one of active component of radix scutellariae, with obvious antitumor action, and being one has wide application
The natural antitumor medicine of prospect.However, oroxylin pharmacological action is extensively, the mechanism of its antitumor action is not yet complete at present
Illustrate.Qroxylin A -7- glucuronides (oroxylin glycosides, Oroxyloside, OAG) are also that the activearm of radix scutellariae divides it
One, with oroxylin not same substance, the research to oroxylin glycosides is less at present, the research of related pharmacological action there is not yet
Report.
The content of the invention
It is an object of the invention to provide application of the oroxylin glycosides in antineoplastic is prepared.
The purpose of the present invention is realized by following technical measures：
Application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared, in particular for the medicine for treating tumor metastasis of liver cancer, its
The mechanism for playing inhibiting effect on tumor metastasis is that oroxylin glycosides has suppression tumor cell invasion and migration for tumour cell
Ability.Its preparation can be any one formulation pharmaceutically allowed, including but not limited to tablet, granule, pill, oral
Liquid, injection, film, capsule, liposome etc..The consumption of oroxylin glycosides can according to route of administration, patient age, body weight,
The change such as body surface area, the disease type treated and the order of severity and course for the treatment of arrangement, its daily dose can be 200-8000mg,
It can be applied with one or many.
Beneficial effect of the present invention：
Oroxylin glycosides is with the ability for preferably suppressing tumor cell invasion and migration, available for preparation anti-tumor metastasis
Medicine, the medicine of the anti-tumor metastasis of various formulations can be prepared into, more preferable selection is provided for tumour patient.
Brief description of the drawings
The influence that Fig. 1 .OAG are migrated to human liver cancer cell HepG2.
The influence that Fig. 2 .OAG are attacked to human liver cancer cell HepG2.
Embodiment
The invention will be further elaborated by the following examples.
Test examples 1：The influence (Cell migration assay) that OAG is migrated to human liver cancer cell HepG2
1st, experiment material
(1) medicine
Qroxylin A -7- glucuronides (oroxylin glycosides, Oroxyloside, OAG) are bought from market, light yellow
Powder.Content：90%, mother liquor is made into DMSO, -20 DEG C of preservations are placed in.It is dense needed for being made into before use with RPMI-1640 nutrient solutions
Degree.
(2) cell line
Human liver cancer cell HepG2 is ground from Shanghai Life Sciences Research Institute, Chinese Academy Of Sciences' biochemistry with cell biology
Institute's cell bank is studied carefully, with the 1640 culture medium culture containing 10% calf serum.
(3) reagent
1) nutrient solution：RPMI-1640 culture mediums, U.S.'s GIBCO Products.RPMI-1640 powder 10.4g is taken to be dissolved in
In 1000mL sterilizing tri-distilled waters, NaHCO is used3PH value is adjusted to 7.3~7.4, cylindric style filter filtration sterilization, packing, 4 DEG C of refrigerators
Preserve.Use 10% hyclone of preceding addition, 100U/mL penicillin and 100mg/L streptomysins.
2) calf serum：Hangzhou Chinese holly bio-engineering corporation product.30min is inactivated through 56 DEG C of water-baths, dispenses and preserves
In -20 DEG C of low temperature refrigerators.
3) PBS：Weigh NaCl 8.0g, KCl 0.20g, Na2HPO4·H2O 1.56g、KH2PO42.0g, is dissolved in
In 1000mL tri-distilled waters, autoclaving, 4 DEG C of refrigerators are preserved.
4) 0.02%EDTA solution：EDTA 20mg are weighed, are dissolved in 100mL PBSs, stirring at low speed, pH value is adjusted
To 7.3~7.4, cylindric style filter filtration sterilization, packing, 4 DEG C of refrigerators are preserved.
5) 0.25% pancreatin：Pancreatin 0.25g is weighed, is dissolved in 100mL PBSs, autoclaving.
6) DMSO solution：U.S. Sigma-Aldrich (St.Louis, Mo) Products
2nd, laboratory apparatus：
1) YJ-875 types Medical purification workbench：Suzhou Decontamination Equipment Plant produces.
2) 3111 type water-jacket typ CO2Incubator：U.S.'s Thermo electron Products.
3) OlympusIX51 types inverted fluorescence microscope：Japanese Olympus Products.
4) electronic balance：Beijing Sai Duolisi instrument systems Co., Ltd product.
5) LD4-2 generic centrifuges：Beijing Medical Centrifugal Machine Factory's product.
6) Centrifuge 5810R types centrifuge：German Eppendorf Products.
3rd, experimental method：
By HepG2 digestive inoculations to 6 orifice plates, culture is to preparing cut during density 90%.With 10 μ l plastics lancet
Head reaches that the hole center of requirement draws a straight line in cell, and straight line answers equal thickness.After having drawn, the cell hiked up is washed with PBS
Fall.Each hole is accordingly added after the culture medium of the glycosides of oroxylin containing various concentrations (50 μM, 100 μM, 200 μM), is put into 37 DEG C of cultures
Case culture.Respectively at 0h, 12h, 24h, 36h, 48h takes pictures to each dosage group cut.Different time points are taken a picture, intuitively
Observation HepG2 is giving the change of oroxylin glycosides migration distance before and after the processing, as a result sees Fig. 1.
4th, experimental result
As seen from Figure 1, oroxylin glycosides can suppress hepatocellular carcinoma H22 migration, and in regular hour and concentration dependant
Property.
Test examples 2：The influence (cell invasion experiment) attacked to human liver cancer cell HepG2
1st, experiment material
(1) medicine
Qroxylin A -7- glucuronides (oroxylin glycosides, Oroxyloside, OAG) are bought from market, light yellow
Powder.Content：90%, mother liquor is made into DMSO, -20 DEG C of preservations are placed in.It is dense needed for being made into before use with RPMI-1640 nutrient solutions
Degree.
(2) cell line
Human liver cancer cell HepG2 is ground from Shanghai Life Sciences Research Institute, Chinese Academy Of Sciences' biochemistry with cell biology
Institute's cell bank is studied carefully, with the 1640 culture medium culture containing 10% calf serum.
(3) reagent
1) nutrient solution：RPMI-1640 culture mediums, U.S.'s GIBCO Products.RPMI-1640 powder 10.4g is taken to be dissolved in
In 1000mL sterilizing tri-distilled waters, NaHCO is used3PH value is adjusted to 7.3~7.4, cylindric style filter filtration sterilization, packing, 4 DEG C of refrigerators
Preserve.Use 10% hyclone of preceding addition, 100U/mL penicillin and 100mg/L streptomysins.
2) calf serum：Hangzhou Chinese holly bio-engineering corporation product.30min is inactivated through 56 DEG C of water-baths, dispenses and preserves
In -20 DEG C of low temperature refrigerators.
3) PBS：Weigh NaCl 8.0g, KCl 0.20g, Na2HPO4·H2O 1.56g、KH2PO42.0g, is dissolved in
In 1000mL tri-distilled waters, autoclaving, 4 DEG C of refrigerators are preserved.
4) 0.02%EDTA solution：EDTA 20mg are weighed, are dissolved in 100mL PBSs, stirring at low speed, pH value is adjusted
To 7.3~7.4, cylindric style filter filtration sterilization, packing, 4 DEG C of refrigerators are preserved.
5) 0.25% pancreatin：Pancreatin 0.25g is weighed, is dissolved in 100mL PBSs, autoclaving.
6) DMSO solution：U.S. Sigma-Aldrich (St.Louis, Mo) Products.
7) haematine；Yihong；Fixer (methanol).
2nd, laboratory apparatus：
1) YJ-875 types Medical purification workbench：Suzhou Decontamination Equipment Plant produces.
2) 3111 type water-jacket typ CO2Incubator：U.S.'s Thermo electron Products.
3) OlympusIX51 types inverted fluorescence microscope：Japanese Olympus Products.
4) electronic balance：Beijing Sai Duolisi instrument systems Co., Ltd product.
5) LD4-2 generic centrifuges：Beijing Medical Centrifugal Machine Factory's product.
6) Centrifuge 5810R types centrifuge：German Eppendorf Products.
7) transwell cells；24 orifice plates.
3rd, experimental method：
Cell suspension sub-bottle agent-feeding treatment.HepG2 is clicked and entered in 6 orifice plates, and when density is 60%, oroxylin is given respectively
(25 μM, 50 μM, 100 μM of glycosides；200 μM), it is put into 48h in cell culture incubator.Dilute matrigel (matrigel：Serum-free is trained
Support base=1：8) matrigels of the 100 μ l to have diluted, 37 DEG C of standing more than 30min, modeling, are added per cell.Digestion is through thousand
The treated HepG2 cells of layer paper factor glycosides, are resuspended with the culture medium of serum-free, are diluted to 2 × 105Individual cell/ml.Take out
Transwell cells are put into 24 orifice bores.Per hole, existing 600 μ l contain the culture medium of serum, and cell upper strata adds cell and hanged
The μ l of liquid 400.It is put into incubator culture 4h.Cell is taken out, the cell not passed through on the inside of miillpore filter is wiped with cotton swab.Fixer is fixed
2 minutes, pure water cleaning.Brazilwood extract dyeing 15 minutes, pure water cleaning；Eosin stains 20 seconds, blow water cleaning, and cotton swab is dried.It is micro-
Five visuals field, take pictures in being taken under mirror above and below left and right, average, and as a result see Fig. 2.
4th, experimental result：
From Figure 2 it can be seen that oroxylin glycosides can suppress hepatocellular carcinoma H22 invasion and attack, and in concentration dependent.
Conclusion：Oroxylin glycosides has the ability for suppressing tumor cell invasion and migration.
Example of formulations 1
Oroxylin glycosides 10g is taken, the appropriate auxiliary material of injection (including freeze drying powder injection and aseptic subpackaged dry powder injection) is added,
Antineoplastic injection is prepared into by injection (including freeze drying powder injection and aseptic subpackaged dry powder injection) technique, every contains thousand layers
Paper factor glycosides 500mg.
Example of formulations 2
Take oroxylin glycosides 10g, add tablet (including slow-release tablet, matrix tablet, coating tablet, dispersible tablet etc.) appropriate auxiliary
Material, is prepared into antineoplastic tablet, every contains thousand by tablet (including slow-release tablet, matrix tablet, coating tablet, dispersible tablet etc.) technique
Layer paper factor glycosides 300mg.
Example of formulations 3
Chinese oroxylin glycosides 10g is taken, the appropriate auxiliary material of capsule is added, antineoplastic capsule is prepared into by capsule technique,
Every 300mg of glycosides containing oroxylin.
Example of formulations 4
Oroxylin glycosides 10g is taken, the appropriate auxiliary material of oral liquid is added, antineoplastic oral liquid is prepared into by oral liquid technique,
Every 300mg containing glycosides containing oroxylin.
Claims (2)
1. application of the oroxylin glycosides in anti-liver cancer and anti-transfer medicine is prepared.
2. application according to claim 1, it is characterised in that described anti-liver cancer and anti-transfer refers to suppress tumor cell invasion
And migration.
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| CN201510114766.7A CN104721210B (en) | 2015-03-16 | 2015-03-16 | Application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared |
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| CN201510114766.7A CN104721210B (en) | 2015-03-16 | 2015-03-16 | Application of the oroxylin glycosides in medicine for treating tumor metastasis is prepared |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1798568A (en) * | 2003-04-04 | 2006-07-05 | 尤尼根制药公司 | Formulation of dual cycloxygenase (cox) and lipoxygenase (lox) inhibitors for mammal skin care |
| CN101244055A (en) * | 2008-03-03 | 2008-08-20 | 中国药科大学 | Application of oroxylin in pharmacy |
| CN101837003A (en) * | 2002-04-30 | 2010-09-22 | 尤尼根制药公司 | Formulation of a mixture of free-B-ring flavonoids and flavans as a therapeutic agent |
-
2015
- 2015-03-16 CN CN201510114766.7A patent/CN104721210B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101837003A (en) * | 2002-04-30 | 2010-09-22 | 尤尼根制药公司 | Formulation of a mixture of free-B-ring flavonoids and flavans as a therapeutic agent |
| CN1798568A (en) * | 2003-04-04 | 2006-07-05 | 尤尼根制药公司 | Formulation of dual cycloxygenase (cox) and lipoxygenase (lox) inhibitors for mammal skin care |
| CN101244055A (en) * | 2008-03-03 | 2008-08-20 | 中国药科大学 | Application of oroxylin in pharmacy |
Non-Patent Citations (1)
| Title |
|---|
| Oroxylin A suppresses invasion through down-regulating the expression of matrix metalloproteinase-2/9 in MDA-MB-435 human breast cancer cells;Yu Sun 等;《European Journal of Pharmacology》;20081210;第603卷;摘要，结果，讨论 * |
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