CN107551011A - A kind of plant extracts for treating metastasis of cancer and preparation method thereof - Google Patents
A kind of plant extracts for treating metastasis of cancer and preparation method thereof Download PDFInfo
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of plant extracts for treating metastasis of cancer and preparation method thereof.Inventor has found first through capturing for many years, confirmation oldenlandia diffusa extract lockable simultaneously can optionally treat metastasis of cancer, oldenlandia diffusa extract reaches 99.8 99.9% to the effective percentage of metastasis of cancer, it is expected to the treatment metastasis of cancer as very attractive, the particularly medicine of brain metastasis and cancer through Lymph Node Metastasis.Possible mechanism of action is by suppressing cell propagation, inducing cell apoptosis and suppressing angiogenesis to play Anticancer metastasis action.There is stronger targeting to treatment metastasis of cancer, drug resistance is not produced, with classic chemotherapy medicine without crossing drug resistant.Oldenlandia diffusa extract is safe and nontoxic, is expected to turn into a kind of new treatment metastasis of cancer, particularly the plant new drug of brain metastasis and cancer through Lymph Node Metastasis.
Description
Technical field
The present invention relates to field of medicaments, and in particular to Plant Extracts and its preparation method and application, more particularly to
Herba hedyotis diffusae extract and its preparation method and application.
Background technology
It is well known that malignant tumour is the frequently-occurring disease and common disease of a kind of serious threat human health, with viral disease,
The three challenges of geriatric disease and referred to as modern medicine, its pathogenesis not yet illustrate completely, and therapeutic effect is also unsatisfactory.
According to incompletely statistics, the research and development of the average annual tumour in the whole world and medical expense are paid wages not low tens billion of, and to country, society and individual bring
Massive losses.
Metastatic carcinoma refers to that tumour cell invades lymphatic vessel from original site, blood vessel or other continue to give birth to by way of being brought at it
It is long, form the tumour with original site tumour same type, this process is referred to as shifting, the tumour formed turn into metastatic tumor or
Metastatic carcinoma.Transfer is the feature of malignant tumour.
Common route of metastasis includes lymphatic metastasis, blood vessel shifts, implantation metastasis etc..Once cancer cell invades lymph
Pipe, can come off to form embolus, or in pipe internal breeding, the swollen thing of formation continuity, but majority is to be drenched by lymphatic vessel into region
Fawn on and formed in lymph node and shifted.
Brain metastasis mainly has the place of several attentions：
1st, pectoralgia：Developed into late period for intermittent secret anguish or vexed pain, lung cancer, tumors invading and pleura cause that the pain increased, lung
On one's deathbed symptom also has peripheral type carcinoma of lung to have pectoralgia, painful shoulder and back, brachialgia and intercostal neuralgia etc. to cancer brain metastes, can typically make disease
People is in great pain, and current clinic can take a little anodyne and carry out pain of alleviation symptom, improve the quality of life of advanced lung cancer patient.
2nd, trachyphonia：It is advanced lung cancer common sympton, is due to that compressing recurrent nerve on the left of diaphragm is invaded and indulged in tumour continuation enlargement,
Cause patient's hoarseness, patient typically reacts without upper airway symptoms or pharyngalgia.
3rd, shortness of breath：Carcinomatous obstruction lung lymph gland or larger bronchus can cause hydrothorax or shortness of breath, patient chest is occurred
Vexed, out of breath or asphyxia, threat to life.
4th, tube chamber is pressurized, generated heat：Tumor-infiltrated phenomena such as causing bronchial obstruction, tumour to form cavity, inflammatory can be caused
Heating paresthesia.
5th, spit blood：Lung cancer development reaches an advanced stage and discontinuity a small amount of bloody sputum repeatedly can occur, and general color and luster is more bright-coloured, occasionally has big
Spitting of blood symptom.
6th, be limited lung cancer pain be advanced lung cancer it is dead before common brain metastasis one kind in symptom on one's deathbed, this carcinous pain
Nerve fibre of the pain mainly from neck, the sensation of domination upper limbs and motion, tumour are once dipped into area and often cause side
It is the pain of upper limbs, weak.Here it is a kind of extremely preceding common sympton of the often advanced lung cancer based on shoulder pain.
7th, face, neck oedema are also the extremely preceding brain metastasis of advanced lung cancer Symptoms on one's deathbed.
The time that general lymphatic metastasis occurs is more early, and its scope may be also more extensive.When the lymph containing cancer cell
(along ductus thoracicus) after into blood, or cancer cell directly invades thin vessels, it is possible to hematogenous metastasis occurs.Into the cancer in blood
Cell moves by individual cells or in the form of by cellulose being linked to be one in blood flow.Generally enter cancer cell in blood circulation not
It can survive, but work as the chance that they are stopped in the process of running, then can invade out tube wall and enter circumvascular interstitial, it is raw
Grow up to transfer stove.Anti-coagulants and chemotherapy are possible to reduce the transfer of tumour, and extrude, partial operation then possible increase transfer
Chance.The different tissue of body has different compatibilities to transfer, and liver, lung, marrow, brain and adrenal gland are common transfer portion
Position, and spleen, muscle etc. then seldom shift.The general more deuterogenesis in the state of an illness of hematogenous metastasis, but lung cancer, breast cancer, kidney
The early stage such as cancer, the cancer of the brain, prostate cancer and thyroid cancer can have hematogenous metastasis.
Metastatic carcinoma is different from primary carcinoma, is because its formation is multifactor, too many levels a continuous dynamic process.
Metastasis cancer cell has more active motility and more powerful drug resistance, particularly possesses abundant new vessels and breeds speed
Degree is greatly accelerated, and has bigger fastness to place side's immunologic function.Treat metastatic carcinoma, it should be Mutiple Targets, be oriented to more
Composite construction medicine.The method of existing treatment metastasis of cancer is extremely limited, less effective.
Oldenlandia diffusa, Chinese medicine name.For Rubiaceae plants of Hedyotis oldenlandia diffusa Hedyotis diffusa Willd.
The herb of [Oldenlandia diffusa (Willd.) Roxb.].Alias：Herba Turczaninowiae Fastigiatae, Radix Picriae felterrae, oldenlandia diffusa..Have
It is clearing heat and detoxicating, carbuncle that disappears dissipating bind, the effect of promoting urination and removing dampness.Herb is very thin, there is most branches, Glabrous, celadon.Single leaf pair
Raw, stockless, linear, full edge, small tooth is arranged at top, and 2, stipule, tiny, tip has small tooth, spend it is tiny, 1-2 it is miscellaneous be born in axil,
Without obstructing or having short stalk, respectively there is a longitudinal furrow capsule nakedness oblate spheroid, both sides, and there are 4 calyx teeth on top.The place of production is in Yunnan, Guangdong, Guangxi, good fortune
Build, Zhejiang, the ground such as Anhui.Harvesting summer and autumn, which uproots, takes herb, picks weeds, shrugs off spare, dries.Meridian distribution of property and flavor：Sweetness and bitterness
It is cold, stomach, large intestine, small intestinl channel are included into, it is nontoxic.For dyspnea and cough due to lung-heat, abscess of throat, acute appendicitis, furuncle sore, venomous snake bite, heat gonorrhea
Puckery pain, oedema, dysentery, enteritis, jaundice with damp-heat pathogen, cancerous swelling.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of eutherapeutic for treating the white of metastasis of cancer
Flower HERBA HEDYOTIS DIFFUSAE extract.
It is another object of the present invention to provide a kind of oldenlandia diffusa alkaloid and its application.
The technical solution used in the present invention is：
A kind of oldenlandia diffusa extract, its preparation method are as follows：
1) oldenlandia diffusa crushed after being dried is taken, adds 5~15 times of water, 40~70 DEG C of leachings are complete；
2) insoluble matter in leachate is removed, is dried to obtain the water extract of oldenlandia diffusa.
As the further improvement of above-mentioned oldenlandia diffusa extract, the insoluble matter in leachate is removed after separation of solid and liquid,
Filtrate then uses 0.45 μm of filter membrane initial filter, is filtered again using 0.22 μm of filter membrane afterwards, further removes insoluble matter.
As the further improvement of above-mentioned oldenlandia diffusa extract, cool drying is used when drying.
Contain 88~95g oldenlandia diffusa alkaloids in per kg oldenlandia diffusa extracts.
A kind of oldenlandia diffusa alkaloid, its preparation method are as follows：
1) above-mentioned oldenlandia diffusa extract is taken, 500~700 DEG C of ashing are complete, obtain calcination；
2) calcination is mixed with water, stirring and dissolving, removes insoluble matter, take filtrate after refined filtration, be dried to obtain oldenlandia
Careless alkaloid.
As the further improvement of above-mentioned oldenlandia diffusa alkaloid, the operation of refined filtration is at the beginning of to use 0.45 μm of filter membrane
Filter, is filtered again using 0.22 μm of filter membrane afterwards.
Oldenlandia diffusa extract is preparing the application in treating metastasis of cancer medicine, wherein, oldenlandia diffusa extract is such as
It is upper described.
A kind of medicine for treating metastasis of cancer, its action composition are above-mentioned oldenlandia diffusa extract.
Medicine is oral formulations.It is common that oral formulations include but is not limited to tablet, granule, capsule, oral liquid etc.
Oral formulations.Further, oral formulations are enteric oral preparation.Particularly, oral formulations are sustained release preparation.
Function is with curing mainly：It is mainly used in treating metastasis of cancer, particularly brain metastasis, cancer is through Lymph Node Metastasis.
Usage and dosage：Freeze-dried powder or spray drying, granule and electuary, standard/grain 500mg, Coming-of-Age Day take 4-5 times,
Four hours clothes/time, one time 5-10 grams, 30 days courses for the treatment of, warm water auf nuechternen Magen einnehmen.
Side effect：Not it was observed that obvious toxic side effect, no drug dependence.
Taboo：Prohibit clothes during pregnant woman and lactation.
The beneficial effects of the invention are as follows：
Inventor has found first through capturing for many years, in 1000 grams of oldenlandia diffusa water extracts (dry weight) containing 88~
95g oldenlandia diffusa alkaloids, there is no pertinent literature to report so far, confirmation oldenlandia diffusa extract (oldenlandia diffusa biology
The mixture of alkali and spreading hedvotis herb polysaccharide) lockable and it can optionally treat metastasis of cancer, particularly brain metastasis and cancer
Through Lymph Node Metastasis, oldenlandia diffusa extract be expected to the treatment metastasis of cancer as very attractive, particularly brain metastasis and
Cancer is through Lymph Node Metastasis medicine.Oldenlandia diffusa extract reaches 99.8-99.9% to treatment brain metastasis effective percentage, or even reaches
100%.Possible mechanism of action is by suppressing cell propagation, inducing cell apoptosis and suppressing angiogenesis to play anticancer turn
Shifting acts on.There is stronger targeting to treatment metastasis of cancer, drug resistance is not produced, with classic chemotherapy medicine without crossing drug resistant.White flower
HERBA HEDYOTIS DIFFUSAE extract is safe and nontoxic, is expected to turn into a kind of plant new drug of new treatment metastasis of cancer.
Oldenlandia diffusa extract has the features such as novelty, Mutiple Targets, too many levels, manifold effect is presented in treatment metastasis of cancer,
Have no toxic side effect etc. and to be furtherd investigate, be likely to become the new drug of the treatment metastasis of cancer with clinical value.
Oldenlandia diffusa extract is a kind of most fast most rapid autonomic drug of current treatment metastasis of cancer, and its appearance will give
Treatment metastasis of cancer, which is brought, once to be broken through.It can save ten hundreds of patient vitals, cytology toxicity test result rapidly simultaneously
It has been shown that, oldenlandia diffusa extract do not have inhibitory action to normal cell, have selective killing effect to cancer cell substantially.
The herba hedyotis diffusae extract of the present invention, there is obvious inhibitory action, its IC to brain metastasis cell propagation50
For KP-N-NS 17.965mg/ml, there is obvious apoptosis-promoting effect to brain metastasis cell, and there is concentration-effect relation；
Can be by the cell-cycle arrest of brain metastasis cell in G0/G1Phase, and there is concentration-effect relation.Can be effective after oral
Treat brain metastasis in ground.
The oldenlandia diffusa alkaloid extracting method of the present invention, without using organic solvent, simple and safe operation, extraction effect
Rate is high, and HPLC testing results show obtained oldenlandia diffusa alkaloid high purity more than 99.9%.
Brief description of the drawings
Fig. 1 is absorbance curve of the oldenlandia diffusa extract aqueous solution under 200nm~600nm wavelength；
Fig. 2 is the liquid chromatogram of oldenlandia diffusa extract；
Fig. 3 is the mass spectrogram of another liquid chromatogram and its main peak of oldenlandia diffusa extract；
Fig. 4 is the HPLC chromatogram of oldenlandia diffusa alkaloid；
Fig. 5 is concentration-inhibiting rate curve of the oldenlandia diffusa extract to KP-N-NS cells；
Fig. 6 is the influence that oldenlandia diffusa extract is bred to human lung cancer (brain metastes) cell KP-N-NS；
Fig. 7 is influence of the various concentrations oldenlandia diffusa extract to human lung cancer (brain metastes) cell KP-N-NS apoptosis；
Fig. 8 is influence of the oldenlandia diffusa extract to human lung cancer (brain metastes) cell KP-N-NS apoptosis；
Fig. 9 is influence of the oldenlandia diffusa extract to human lung cancer (brain metastes) the cell KP-N-NS cell cycles.
Embodiment
Oldenlandia diffusa extracts the preparation of thing：
1) by 55~65 DEG C of crushed after being dried of oldenlandia diffusa to 80~120 mesh, the water of 5~15 times of addition, in 45~55
Stirring extraction 4h at DEG C；
2) insoluble matter is removed, using 0.45 μm of filter membrane initial filter, is filtered afterwards using 0.22 μm of filter membrane two, collects what is obtained
Filtrate is freeze-dried, and obtains oldenlandia diffusa extract.
Above-mentioned oldenlandia diffusa extract is taken to be detected, testing result is as shown in Figures 1 to 3.Wherein, Fig. 1 is white flower
The HERBA HEDYOTIS DIFFUSAE extract aqueous solution (300mg/mL), the absorbance curve under 200nm~600nm wavelength, shows it in 282.5nm
Place has a peak value, and its absorbance is up to 0.981；Fig. 2 is the liquid chromatogram of oldenlandia diffusa extract；Fig. 3 is long-noded pit viper
The mass spectrogram of another liquid chromatogram and its main peak of tongue grass extract.
After testing, 88~95g oldenlandia diffusa alkaloids are contained in every 1000g oldenlandia diffusas extract (dry weight), it is remaining
Measure the impurity for spreading hedvotis herb polysaccharide and very small amount (being less than 0.5%).
The extraction of oldenlandia diffusa alkaloid：
1) foregoing extract (containing 88~95g oldenlandia diffusas alkaloid in per 1000g dry weights) is taken, 500 DEG C~700
DEG C 2~3h of high-temperature digestion, obtains the grey powder of charged material weight 30~35% or so；
2) grey powder is mixed with 3~4 times of water, stirring and dissolving, removes insoluble matter, obtain filtrate；
3) 0.45 μm of filter membrane initial filter is used, is filtered again using 0.22 μm of filter membrane afterwards, obtains refined filtration liquid；
4) refined filtration liquid is freeze-dried to obtain oldenlandia diffusa alkaloid.
Oldenlandia diffusa alkaloid is the transparent needle crystals of pure white, and commission Shanghai Zhangjiang medicine-valley public service platform has
Its purity detects in limit company, and test basis are Chinese Pharmacopoeia 2010 edition one, annex VID high performance liquid chromatographies.Its HPLC color
For spectrogram as shown in figure 4, according to area normalization method, its main peak content is 99.94%, is a kind of high-purity extract.
With reference to experiment, technical scheme is further illustrated.
In testing below, XL-007 refers to the drying oldenlandia diffusa extract that the above method is prepared.
Autonomic drug XL-007 brain metastasis cell growth inhibition check experiments
Experiment commission Hunan Province's Experimental Animal Center (Drug Safety Evaluation Center of Hunan Province) is carried out.
1 experiment material
1.1 tested material：Autonomic drug is numbered：XL-007.Autonomic drug is prepared：XL-007 9g are weighed, add 0.9% sodium chloride
Parenteral solution 15ml, and vibrate to whole dissolvings, 600mg/ml plant drug solns are produced, this is maximum concentration working solution, by mother liquor
It is standby after progress bacteria removing.Mother liquor is configured to 200 successively with 0.9% sodium chloride injection, 60,20,6,2,0.6mg/ml
Working solution.
1.2 positive control drug：Cis-platinum (DDP), lot number：SJJMI-IE, Tokyo HuaCheng Industry Co., Ltd；5 FU 5 fluorouracil
(5-FU), lot number：HFBM160120325008, Amresco company.Positive control drug is prepared：DDP or 5-FU2mg is weighed, with new
Fresh complete medium is configured to 100mM mother liquor, then mother liquor is configured to fresh complete medium 200 successively, 60,20,6,
2nd, 0.6 μM of working solution.
1.3 main material：
1.4 key instrument：
2 test methods
2.1 cell culture
The KP-N-NS cells covered with are taken, using the DMEM in high glucose complete medium containing 10%FBS, in 37 DEG C, 5%CO2
Cultivated in incubator, according to cell growth status, 1~2d is passed on or changed liquid, standby to exponential phase.
2.2CCK-8 methods detect cell proliferation test
Take the logarithm the phase growth KP-N-NS cells with every hole 5 × 103Individual cell is inoculated in 96 porocyte culture plates, is treated
After 12h cell attachments, vehicle control group, positive control drug (DDP) group, positive control drug (5-FU) group, autonomic drug XL- are set
007 group (0.3-300mg/ml), every group of 5 multiple holes.Vehicle control group is with fresh DMEM culture medium incubated cells completely, autonomic drug
Group respectively with the fresh DMEM incubated cells completely containing final concentration of 0.3-300mg/ml autonomic drugs, DDP and 5-FU groups respectively with
Fresh DMEM incubated cells completely containing final concentration of 100,30,10,3,1,0.3 μM of autonomic drugs.It is incubated by above-mentioned processing mode
The μ l of CCK-8 10 are added after cell 72h in every hole, continue to cultivate the extinction for using ELIASA to measure each hole at 450nm after 1h
Degree.Using vehicle control group OD values as 100% cell viability, the ratio of remaining each group OD values and vehicle control group OD values is relatively living
Power.Toxicity of the autonomic drug to KP-N-NS cells is evaluated with cell proliferation inhibition rate, if having cell proliferation inhibition rate ＞
When 100%, the systematic error of instrument is judged to, based on 100%.
The detection of 2.3 Apoptosis
2.3.1Annexin the double dye method detection Apoptosis of V-FITC and PI
The KP-N-NS cells in exponential phase are taken, conventional digestion collects cell, with 5 × 105The density inoculation in/hole
Into 6 orifice plates, after cultivating 12h, vehicle control group, autonomic drug XL-007 groups (10,30,100mg/ml) are set, and every group 5 multiple
Hole.Vehicle control group with fresh DMEM culture medium incubated cells completely, autonomic drug group respectively with containing final concentration of 10,30,
The fresh DMEM incubated cells completely of 100mg/ml autonomic drugs.After 6h, conventional digestion collects cell, adds 500 μ l Binding
Cell is resuspended in Buffer buffer solutions, and cell is transferred in 1.5ml EP pipes, adds 5 μ l Annexin V-FITC, 5 μ l PI,
15min is incubated under the conditions of room temperature lucifuge, with flow cytomery apoptosis situation.
2.3.2 fluorescence colour detects Apoptosis
Cell is handled according to 2.3.1 methods, after autonomic drug handles 6h, adds Hoechst 33342 per hole in 6 orifice plates
Dyeing liquor 1ml, fully covers cell, is placed in 37 DEG C of culture 20-30min.Dyeing liquor is discarded, glimmering after being washed 2-3 times with PBS
Fluoroscopic examination is carried out under light microscope.
Influence of the 2.4 Flow cytometry autonomic drugs to human lung cancer (brain metastes) the cell KP-N-NS cycles
The KP-N-NS cells in exponential phase are taken, conventional digestion collects cell, with 5 × 105The density inoculation in/hole
Into 6 orifice plates, after 12h cell attachments, vehicle control group, autonomic drug XL-007 groups (100,30,10mg/ml) be set, every group 5
Individual multiple holes.Vehicle control group with fresh DMEM culture medium incubated cells completely, autonomic drug group respectively with containing final concentration of 10,30,
The fresh DMEM incubated cells completely of 100mg/ml autonomic drugs.After 6h, conventional digestion collects cell, is fixed with 70% cold ethanol
At night, add 5 μ l PI, room temperature lucifuge is incubated 30min, with the flow cytomery cell cycle.
2.5 statistical analysis
Data are handled using the statistical softwares of SPSS 16.0, measurement data withRepresent, the ratio of two sample averages
Examined compared with using Student T-Test, the comparison of mean is using One-way ANOVA inspections, P between multisample group<0.05 represents
It is statistically significant, P<0.01 represents that examined difference is very significant.
Evaluation of result
The influence that 3.1 autonomic drug XL-007 breed to human lung cancer (brain metastes) cell KP-N-NS
After the autonomic drug processing cell of micro- Microscopic observation various concentrations, there is cell proliferation rate and slow down, cell fragment
Increase, space between cells increases, phenomena such as shape of sand vacuole occurs in cell.Incubation time has clear and definite correlation to cell state together,
About after 12h is co-cultured, being rounded occurs in cell, the phenomenon of shrinkage；After 24h is co-cultured, there is part cell and swell, cell is saturating
Photosensitiveness is deteriorated, intercellular gap increase；After 48h is co-cultured, there is shape of sand vacuole in cell, the situations such as cell rupture occurs；
After 72h is co-cultured, shape of sand vacuole like cell substantially completely ruptures, without complete under 300,100,30mg/ml concentration conditions
The cell of cellular morphology, only see and be condensed into stain shape on a small quantity.Cell co-cultures with test sample or positive control drug DDP and 5-FU
After 72h, cell propagation is substantially suppressed, and has concentration-effect relation, with significant difference compared with vehicle control group
(P<0.01).Cell inhibitory effect is the results detailed in Table 1.
The influence that table 1, autonomic drug XL-007 breed to human lung cancer (brain metastes) cell KP-N-NS
Note：* represent compared with vehicle control group, P<0.05, * * expressions are compared with vehicle control group, P<0.01.IC50Represent
Suppress the concentration of 50% tumour cell, Emax represents the maximal percentage inhibition to tumour cell.
XL-007 is as shown in Figure 5 to concentration-inhibiting rate curve of KP-N-NS cells.
Autonomic drug XL-007 shows that human lung cancer (brain metastes) cell KP-N-NS influence such as Fig. 6 bred arrow represents necrosis
Cell.It can be seen that autonomic drug XL-007 can promote human lung cancer (brain metastes) cell KP-N-NS downright bad well.
Influences of the 3.2 autonomic drug XL-007 to human lung cancer (brain metastes) cell KP-N-NS apoptosis
Use concentration for 10,30, after 100mg/ml autonomic drug intervenes human lung cancer (brain metastes) cell KP-N-NS 6h, receive
Collect cell, Apoptosis is detected after the double dyes of Annexin V-FITC/PI.Cell after double dyes can be divided into 4 groups by flow cytometer：
Q1-UL represents mechanical damage cell (Annexin V-/PI+)；Q1-UR non-viable apoptotic cells (Annexin V+/PI+)；Q1-LL
Survivaling cell (Annexin V-/PI-)；Q1-LR viable apoptotic cells (Annexin V+/PI-).
The influence of table 2, autonomic drug XL-007 to human lung cancer (brain metastes) cell KP-N-NS apoptosis
Note：* represent compared with vehicle control group, P<0.05, * * expressions are compared with vehicle control group, P<0.01.
As a result show：Through autonomic drug XL-007 handle KP-N-NS cells late apoptic and non-viable non-apoptotic cell number apparently higher than
Vehicle control group (P<0.05), and there is concentration-effect relation.
Influences of the various concentrations autonomic drug XL-007 to human lung cancer (brain metastes) cell KP-N-NS apoptosis is as shown in fig. 7, figure
In, A is vehicle control group, and B is autonomic drug 10mg/ml groups, and C is autonomic drug 30mg/ml groups, and D is autonomic drug 100mg/ml groups.
Influences of the autonomic drug XL-007 to human lung cancer (brain metastes) cell KP-N-NS apoptosis is as shown in figure 8, arrow represents thin
Karyon shrinkage, prompts for apoptotic cell.It can clearly be seen that XL-007 can promote human lung cancer (brain metastes) cell from figure
KP-N-NS apoptosis.
Influences of the 3.3 autonomic drug XL-007 to human lung cancer (brain metastes) the cell KP-N-NS cycles
Use concentration for 10,30, after 100mg/ml autonomic drug intervenes human lung cancer (brain metastes) cell KP-N-NS 6h, receive
Collect cell, after being fixed with 70% cold ethanol, after PI dyeing, the flow cytometry analysis cell cycle.Experimental result is as shown in table 3：
The influence of table 3, autonomic drug XL-007 to human lung cancer (brain metastes) the cell KP-N-NS cell cycles
Note：* represent compared with vehicle control group, P<0.05, * * expressions are compared with vehicle control group, P<0.01
As a result show：After using autonomic drug XL-007, G0/G1Cell proportion substantially increases, G2/ M phase ratios substantially subtract
It is few, show it to human lung cancer (brain metastes) cell KP-N-NS inhibited proliferations mainly by human lung cancer (brain metastes) cell KP-
N-NS is arrested in G0/G1Phase, it is prevented to enter the S phases.
Influences of the autonomic drug XL-007 to human lung cancer (brain metastes) cell KP-N-NS cell cycles is as shown in figure 9, in figure, A
For vehicle control group, B is autonomic drug 10mg/ml groups, and C is autonomic drug 30mg/ml groups, and D is autonomic drug 100mg/ml groups.
4 conclusions are with discussing
In summary, under this experiment condition, autonomic drug XL-007 can significantly inhibit a kind of human lung cancer (brain metastes) cell
KP-N-NS breeds, and has concentration-effect relation, under a high concentration condition, with the extension of time, can be complete by cancer cell
Complete to kill, it can be by cell-cycle arrest in G0/G1Phase, inducing cell apoptosis.This autonomic drug play suppress cancer cell concentration compared with
Height, it is mg/ml levels, may be related to its distinctive mechanism of action.
Described above is only the preferred embodiment of the present invention, it is noted that for the common skill of the art
For art personnel, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications
Also within protection scope of the present invention.
Claims (10)
1. a kind of oldenlandia diffusa extract, its preparation method are as follows：
1) oldenlandia diffusa crushed after being dried is taken, adds 5~15 times of water, 40~70 DEG C of leachings are complete；
2) insoluble matter in leachate is removed, is dried to obtain the water extract of oldenlandia diffusa.
2. oldenlandia diffusa extract according to claim 1, it is characterised in that：Removed after separation of solid and liquid in leachate
Insoluble matter, filtrate are then used 0.45 μm of filter membrane initial filter, filtered again using 0.22 μm of filter membrane afterwards, are further removed not
Molten thing.
3. oldenlandia diffusa extract according to claim 1 or 2, it is characterised in that：Cool drying is used when drying.
4. oldenlandia diffusa extract according to claim 1 or 2, it is characterised in that：Per kg oldenlandia diffusa extracts
In contain 88~95g oldenlandia diffusa alkaloids.
5. a kind of oldenlandia diffusa alkaloid, its preparation method are as follows：
1) the oldenlandia diffusa extract described in claims 1 to 33 is taken, 500~700 DEG C of ashing are complete, obtain calcination；
2) calcination is mixed with water, stirring and dissolving, removes insoluble matter, filtrate is taken after refined filtration, be dried to obtain oldenlandia diffusa life
Alkaloids.
A kind of 6. oldenlandia diffusa alkaloid according to claim 5, it is characterised in that：The operation of refined filtration is to use 0.45
μm filter membrane initial filter, filtered again using 0.22 μm of filter membrane afterwards.
7. oldenlandia diffusa extract is preparing the application in treating metastasis of cancer medicine, it is characterised in that：Oldenlandia diffusa extracts
Thing is as described in any one of claims 1 to 3.
A kind of 8. medicine for treating metastasis of cancer, it is characterised in that：Its action composition is white described in any one of Claims 1 to 4
Flower HERBA HEDYOTIS DIFFUSAE extract.
9. medicine according to claim 8, it is characterised in that：Medicine is oral formulations.
10. medicine according to claim 9, it is characterised in that：Metastasis of cancer is selected from brain metastasis, cancer Lymph Node Metastasis.
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| 李洁等: "白花蛇舌草抑制Lewis肺癌小鼠自发转移的实验研究", 《中医药导报》 * |
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