CN107693528A - Ginseng saponin Rh 2 promotes the new application that medicine penetrates in solid tumor - Google Patents
Ginseng saponin Rh 2 promotes the new application that medicine penetrates in solid tumor Download PDFInfo
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- CN107693528A CN107693528A CN201710949831.7A CN201710949831A CN107693528A CN 107693528 A CN107693528 A CN 107693528A CN 201710949831 A CN201710949831 A CN 201710949831A CN 107693528 A CN107693528 A CN 107693528A
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- solid tumor
- adriamycin
- ginseng saponin
- medicine
- promotes
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- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 34
- 241000208340 Araliaceae Species 0.000 title claims abstract description 25
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 title claims abstract description 25
- 235000003140 Panax quinquefolius Nutrition 0.000 title claims abstract description 25
- 235000008434 ginseng Nutrition 0.000 title claims abstract description 25
- 239000001397 quillaja saponaria molina bark Substances 0.000 title claims abstract description 23
- 229930182490 saponin Natural products 0.000 title claims abstract description 23
- 150000007949 saponins Chemical class 0.000 title claims abstract description 23
- 239000003814 drug Substances 0.000 title claims abstract description 18
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims abstract description 44
- 229940009456 adriamycin Drugs 0.000 claims abstract description 22
- 230000000694 effects Effects 0.000 claims abstract description 6
- 230000000149 penetrating effect Effects 0.000 claims abstract description 6
- 238000009826 distribution Methods 0.000 claims abstract description 4
- 206010006187 Breast cancer Diseases 0.000 claims description 9
- 208000026310 Breast neoplasm Diseases 0.000 claims description 9
- 230000000118 anti-neoplastic effect Effects 0.000 claims description 8
- 241001465754 Metazoa Species 0.000 abstract description 8
- 230000007774 longterm Effects 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 4
- 241000050051 Chelone glabra Species 0.000 abstract description 3
- 230000001413 cellular effect Effects 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 15
- 241000699660 Mus musculus Species 0.000 description 5
- 238000011580 nude mouse model Methods 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 238000003304 gavage Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000012679 serum free medium Substances 0.000 description 4
- 210000003462 vein Anatomy 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 235000010603 pastilles Nutrition 0.000 description 2
- 229960004964 temozolomide Drugs 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 208000019065 cervical carcinoma Diseases 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 230000027288 circadian rhythm Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- OORMXZNMRWBSTK-LGFJJATJSA-N dammarane Chemical group C1CCC(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@H]([C@H](C)CCCC(C)C)[C@H]4CC[C@@H]3[C@]21C OORMXZNMRWBSTK-LGFJJATJSA-N 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000009513 drug distribution Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 230000003014 reinforcing effect Effects 0.000 description 1
- 238000012764 semi-quantitative analysis Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 210000005166 vasculature Anatomy 0.000 description 1
Classifications
-
- 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
Abstract
The new application that the present invention penetrates for research ginseng saponin Rh 2 promotion medicine in solid tumor.This technology invention belongs to natural medicine field, is related to the new application that the active skull cap components ginseng saponin Rh 2 of Chinese medicine ginseng promotes medicine to be penetrated in solid tumor.Simulate in vitro and study and confirm the new application that ginseng saponin Rh 2 promotes medicine to be penetrated in solid tumor on the sphere cells model and the Transplanted tumor model that whole animal is horizontal of solid tumor.Main research is as follows：Based on sphere cells model, single combination and the long-term influence penetrated in advance to Rh2 to adriamycin are investigated.Whole animal it is horizontal investigate single combination and it is long-term in advance to Rh2 after, penetrate behavior in the knurl of adriamycin.The present invention with entirety and cellular pharmacokinetics research meanses has found and demonstrated that adriamycin penetrating in sphere cells model and solid tumor can be promoted to Rh2 in advance for a long time, and Rh2 is obvious in central area effect, heterogeneity distribution few in more outside adriamycin is improved.
Description
Technical field
The present invention relates to natural medicine field, and in particular to the active skull cap components ginseng saponin Rh 2 of Chinese medicine ginseng promotes
Medicine penetrated in solid tumor in new application.
Background technology
At present, tumour is still to threaten the major disease of human health, and one of Global mortality highest disease.2017
The data of year newest issue of National Cancer Center show that about 10,000 people make a definite diagnosis cancer daily in China, and per minute just to there are about 7 people true
Examine and suffer from cancer.So far, there has been no the effect method for the treatment of tumour, chemotherapy to remain one of Main Means of oncotherapy.Grind
Study carefully and show, antineoplastic enter body after by metabolism disposal process mainly include：Integral level, tissue level and cell
It is horizontal.Wherein absorption, distribution, metabolism and the excretion of antineoplastic integral level can be divided by classical pharmacokinetics
Analysis；It can be investigated in the intake of cellular level and subcellular redistribution by target cell pharmacokinetics；But in tissue water
In flat, antineoplastic passes through penetrating so as to reach this process of each region cell in knurl often by people in tumor tissues
Ignore.Meanwhile antineoplastic penetrates process by many limitations in knurl, including abnormal tumor vasculature,
Fine and close extracellular matrix and the Fluid pressure in tumor tissues gap etc..Therefore, finding one kind can be with antineoplastic Internet of Things
With it is very necessary to improve the compound that antineoplastic penetrates in knurl.
Ginseng is herbaceos perennial, sweet, slight bitter, warm-natured, flat.With reinforcing vital energy, veins takes off admittedly, tonifying spleen benefit
Lung, promote the production of body fluid, calm the nerves and other effects.Life saponin(e Rh2 is isolated active skull cap components from ginseng, and its content is extremely low, carries
It is only 0.001% to take rate.Ginseng saponin Rh 2 has dammarane's structure, belongs to protopanaxadiol-type's saponin(e, the entitled protoplast's ginseng of chemistry
Glycol -3- oxygen-B-D- glucopyranosides, have many pharmacological activity, enjoy the good reputation of " shield life element ".Its structural formula is as follows：
Patent CN104337823A discloses a kind of pharmaceutical composition for suppressing tumour, be it is a kind of with ginseng saponin Rh 2 and
Temozolomide is the compound medicinal formulation of active constituents of medicine, in range of doses, ginseng saponin Rh 2 and Temozolomide connection
Share medicine has the function that Synergistic to glioma and cervical carcinoma.Patent CN104232584A is disclosed in multi-layer cellular mould
In type, ginseng saponin Rh 2 is combined with adriamycin, and the intracellular intake and transcellular transport that can increase antineoplastic are realized and reversed
Tumor drug resistance.However, above-mentioned patent is not directed to the effect that ginseng saponin Rh 2 promotes medicine to be penetrated in solid tumor.Therefore, base
In above-mentioned reason, we simulate in vitro grinds on the sphere cells model and the Transplanted tumor model that whole animal is horizontal of solid tumor
Study carefully and confirm the new application that ginseng saponin Rh 2 promotes medicine to be penetrated in solid tumor.
The content of the invention
The purpose of the present invention is the transplantable tumor for the sphere cells model and whole animal level for simulating solid tumor in vitro
The new application that ginseng saponin Rh 2 promotes medicine to be penetrated in solid tumor is studied and confirmed on model.
The research approach of the present invention is that (1) investigates single combination with long-term in advance to ginseng based on breast cancer sphere cells model
The influence that saponin(e Rh2 confrontation anti-neoplastic drug doxorubicins penetrate.(2) it is combined in the horizontal investigation single of whole animal and gives people in advance for a long time
After joining saponin(e Rh2, behavior is penetrated in the knurl of adriamycin.
The result of study of the present invention, which mainly includes single combination ginseng saponin Rh 2, can significantly improve adriamycin in breast cancer
Penetrating in sphere cells model and solid tumor, but Rh2 effect is mainly in outer region.And give ginseng saponin Rh 2 in advance for a long time
Adriamycin penetrating in breast cancer sphere cells model and solid tumor can be significantly improved, and Rh2 is bright in central area effect
It is aobvious, the drug distribution of central area and outer region is reached unanimity, it is existing to improve heterogeneity distribution few in more outside adriamycin
As.
Brief description of the drawings
Fig. 1；After A, single combination Rh2, Confocal Images that adriamycin penetrates in breast cancer sphere cells model and glimmering
The sxemiquantitative image of luminous intensity.B, it is long-term in advance to the common focused view that after Rh2, adriamycin penetrates in breast cancer sphere cells model
The sxemiquantitative image of picture and fluorescence intensity.
Fig. 2：After A, single combination Rh2, Confocal Images and the semidefinite of fluorescence intensity that adriamycin penetrates in solid tumor
Spirogram picture.B, after giving Rh2 in advance for a long time, Confocal Images and the semidefinite spirogram of fluorescence intensity that adriamycin penetrates in solid tumor
Picture.
Embodiment
Embodiment 1：It is combined based on breast cancer sphere cells model investigation single and adriamycin is penetrated to Rh2 in advance for a long time
It is as follows to influence key step：
Be divided into three groups, respectively control group, single combination group, give group in advance for a long time.(1) control group：The mammary gland that will be established
Cancer sphere cells model (7.5 × 103The density in individual/hole, with 96 U-shaped board cultures of SUMILON 7 days) it is transferred to sterile 5ml
In EP pipes (12 ball/pipes), serum free medium 2ml is added, in 37 DEG C, containing 5%CO2, in the incubator of relative humidity 90%
After being incubated 72h, culture medium is discarded.Then add the serum free medium 2ml containing 5 μM of adriamycins, respectively at administration 1,4,8,16,
After 24h, pastille culture medium is removed, PBS is washed 3 times, and Z axis scanning is carried out with 10 μm of intervals under Laser Scanning Confocal Microscope.(2) single
Combination group：By the breast cancer sphere cells Model transfer established into sterile 5ml EP pipes (12 ball/pipes), no blood is added
Clear culture medium 2ml, in 37 DEG C, containing 5%CO2, after being incubated 71h in the incubator of relative humidity 90%, award the nothing containing 5 μM of Rh2
Blood serum medium 2ml is incubated 1h, not withdrawal, is further continued for awarding 5 μM of adriamycins, after adriamycin 1,4,8,16,24h is awarded, removes
Pastille culture medium is gone, PBS is washed 3 times, and Z axis scanning is carried out with 10 μm of intervals under Laser Scanning Confocal Microscope.(3) group is given in advance for a long time：
By the breast cancer sphere cells Model transfer established into sterile 5ml EP pipes (12 ball/pipes), add containing 5 μM of Rh2
Serum free medium 2ml, in 37 DEG C, containing 5%CO2, after being incubated 72h in the incubator of relative humidity 90%, discard drug containing culture
Base.The serum free medium 2ml containing 5 μM of adriamycins is awarded again, after administration 1,4,8,16,24h, removes drug containing culture
Base, PBS are washed 3 times, and Z axis scanning is carried out with 10 μm of intervals under Laser Scanning Confocal Microscope.530~560nm of exciter filter, transmitting
573~647nm of optical filter.Progress fluorescence semi-quantitative analysis at maximum gauge is taken, fluorescence intensity calculates with Zen softwares.
Embodiment 2：Based on the horizontal influence master for investigating single combination and being penetrated in advance to Rh2 to adriamycin for a long time of whole animal
Want step as follows：
Female BAl BIc/c nude mices (18~22g, 8~10 week old) of health are purchased from the limited public affairs of Shanghai Si Laike experimental animals
Department.Nude mice is raised in SPF level Animal Houses, and rearing conditions are 5/cage, room temperature (22 ± 1 DEG C), 50~60% relative humidity, 12h
Circulate circadian rhythm.5 × 10 are subcutaneously injected on the right side of nude mice6Individual exponential phase MCF-7 cells.Tumor mouse model is established
Afterwards, tumour growth situation is observed, it is random by body weight and knurl volume after the heterologous graftings of MCF-7 reach 8~10mm of average diameter
It is divided into three groups, every group 10.All animals fasting 12h before last time is administered, but can be with free water.(1) control group：It is naked
The daily gavage of mouse gives 0.5%CMC-Na (0.1ml/10g), altogether be administered 7 days, in the 8th day again tail vein give 30mg/kg Ah
Mycin (3mg/mL, 0.1ml/10g), a length of 30min during administration.(2) single combination group：The daily gavage of nude mice awards 0.5%
CMC-Na (0.1ml/10g), it is administered 7 days altogether, 50mg/kg (5mg/mL, 0.1ml/10g) Rh2,2h is awarded in the 8th day gavage
Tail vein gives 30mg/kg adriamycin (3mg/mL, 0.1ml/10g) again afterwards, a length of 30min during administration.(3) give in advance for a long time
Group：The daily gavage of nude mice awards 50mg/kg (5mg/mL, 0.1ml/10g) Rh2, is administered 7 days altogether, in the 8th day, tail vein was given again
Give 30mg/kg adriamycin (3mg/mL, 0.1ml/10g), a length of 30min during administration.When terminal is administered, plucks eyeball and take at blood
Extremely, tumor tissue and is taken out immediately, is cleaned with physiological saline, and is dried with filter paper, and OCT glue coating, -80 DEG C quick-frozen.Then, ice is used
Freeze slicer to be cut into slices, thickness is 10 μm, and is attached on slide.The red fluorescence of adriamycin uses Laser Scanning Confocal Microscope
Observed (530~560nm of exciter filter, 573~647nm of transmitting optical filter).Fluorescence intensity carries out half with Zen softwares
It is quantitative.
Claims (2)
1. ginseng saponin Rh 2 can promote medicine penetrating in solid tumor.
2. as claimed in claim 1, ginseng saponin Rh 2 promotes penetrating for antineoplastic, it is characterised in that：Single is combined people
The effect that ginseng saponin(e Rh2 promotes adriamycin to penetrate is confined to the outer region of breast cancer sphere cells model and solid tumor, and grows
The central area that acts on that phase promotes adriamycin to penetrate to ginseng saponin Rh 2 in advance becomes apparent, and improves few in more outside adriamycin
Heterogeneity distribution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710949831.7A CN107693528A (en) | 2017-10-09 | 2017-10-09 | Ginseng saponin Rh 2 promotes the new application that medicine penetrates in solid tumor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710949831.7A CN107693528A (en) | 2017-10-09 | 2017-10-09 | Ginseng saponin Rh 2 promotes the new application that medicine penetrates in solid tumor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107693528A true CN107693528A (en) | 2018-02-16 |
Family
ID=61185050
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710949831.7A Pending CN107693528A (en) | 2017-10-09 | 2017-10-09 | Ginseng saponin Rh 2 promotes the new application that medicine penetrates in solid tumor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107693528A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232584A (en) * | 2014-09-15 | 2014-12-24 | 中国药科大学 | Building of in-vitro three-dimensional cell model of breast cancer and application of in-vitro three-dimensional cell model in research of drug resistance mechanism and reversal agents screening |
| CN106997423A (en) * | 2016-07-22 | 2017-08-01 | 中国药科大学 | A kind of structure of medicine penetration PK model based on three-dimensional cell model and its application in drug evaluation |
-
2017
- 2017-10-09 CN CN201710949831.7A patent/CN107693528A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232584A (en) * | 2014-09-15 | 2014-12-24 | 中国药科大学 | Building of in-vitro three-dimensional cell model of breast cancer and application of in-vitro three-dimensional cell model in research of drug resistance mechanism and reversal agents screening |
| CN106997423A (en) * | 2016-07-22 | 2017-08-01 | 中国药科大学 | A kind of structure of medicine penetration PK model based on three-dimensional cell model and its application in drug evaluation |
Non-Patent Citations (1)
| Title |
|---|
| 朴丽花: "人参皂苷Rh2以人乳腺癌细胞多药耐药的逆转作用及其相关机制的实验研究", 《延边大学博士学位论文》 * |
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Application publication date: 20180216 |