CN105232565B - The application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared - Google Patents
The application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared Download PDFInfo
- Publication number
- CN105232565B CN105232565B CN201510498383.4A CN201510498383A CN105232565B CN 105232565 B CN105232565 B CN 105232565B CN 201510498383 A CN201510498383 A CN 201510498383A CN 105232565 B CN105232565 B CN 105232565B
- Authority
- CN
- China
- Prior art keywords
- triterpene glucoside
- pharmaceutically acceptable
- acceptable salt
- triterpene
- tumour radiotherapy
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0038—Radiosensitizing, i.e. administration of pharmaceutical agents that enhance the effect of radiotherapy
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/42—Cucurbitaceae (Cucumber family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Mycology (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Steroid Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides triterpene glucoside or a kind of new application of its pharmaceutically acceptable salt, tumour radiotherapy sensitizer can be prepared with it.It during using being applied to tumour radiotherapy as the tumour radiotherapy sensitizer that active component is prepared by triterpene glucoside and its pharmaceutically acceptable salt, can not only increase the radiosusceptibility of tumour, and the side effect of radiotherapy can also be reduced.Meanwhile triterpene glucoside is nontoxic, human normal cell will not be injured, therefore the present invention has potential applicability in clinical practice.
Description
Technical field
The present invention relates to the application field of triterpene glucoside, in particular to a kind of triterpene glucoside or its pharmaceutically
Application of the acceptable salt in tumour radiotherapy sensitizer is prepared.
Background technology
Tumour includes malignant tumour and benign tumour.Benign tumour be able to can typically be cured by surgery excision, to human body
Harm is smaller.But malignant tumour harm to the human body is larger, it is high to turn into China's incidence of disease height, fatal rate for malignancy disease at present
Disease.At present, radiotherapy, chemotherapy, surgery excision are treatment malignant tumour main methods.Radiotherapy experienced the hair of over one hundred year
Exhibition, turns into a kind of ripe and effectively treatment tumor disease treatment means, malignant tumor patient more than half in the state of an illness not
Need to make radiotherapy with the stage.
Although the therapeutic effect of Radiotherapy in Malignant is reliable, adverse reaction is clear and definite, and most of tumour can be made to obtain not
With the control of degree, but there is also defect.
First, radiotherapy also can more or less damage certain amount while killing or killing malignant cell
Normal cell, and cause some infringements and reaction topically or systemically occur.I.e. using radiation means treatment malignant tumour
During some side reactions, such as local organization swelling, dermohemia, pigmentation occurs, follicular keratosis, skin dryness take off
Skin, alopecia etc.；There is weak, nauseous, anorexia, insomnia, leucocyte decline etc. in whole body.
Second, only radiosensitive malignant tumour has the effect of relatively good to single radiotherapy means.But many evils
Property tumour to radiation and it is insensitive.Now, although increase radiological dose is advantageous to the sensitiveness of increase radiation, but also can be a large amount of
Normal cell is killed or killed, unsurmountable injury is brought to patient.Therefore, when malignant tumour is to radiating insensitive,
Radiotherapy means can only be abandoned.
Triterpene glucoside is a kind of a variety of triterpene compounds extracted from the fruit of cucurbitaceous plant Momordica grosvenori
General name.It not only has the characteristic flavor on basis of Momordica grosvenori, and is used as triterpene glucoside, and it has very high sugariness, its
Sugariness is about 300 times of sucrose, and does not produce heat.Therefore, triterpene glucoside is used frequently as the food additives such as sweetener, essence
Add agent to be applied to the industries such as beverage, candy, food, improve the taste and flavor of product.As active component preparing
Tumour radiotherapy sensitizer, have no that its correlation is recorded so far.
In view of this, it is special to propose the present invention.
The content of the invention
It is an object of the invention to provide triterpene glucoside or its pharmaceutically acceptable salt to prepare tumor chemotherapy sensitizing
Application in agent.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
The application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared.
Using radiation means treatment malignant tumour when, some side effects usually occur, or the malignant tumour having for
Radiate it is insensitive, so as to can not be treated using radiation means.The present invention has found that triterpene glucoside has well by studying
Tumor radiotherapy enhanced sensitivity effect, therefore can be used to prepare radiosensitizer.So far there is not yet on triterpene glucoside this
The report of application.
Preferably, the triterpene glucoside has following structure formula：
In formula, R and R1For glucose residue, the chemical constitution of the glucose residue includes
In any one.
Preferably, the triterpene glucoside is saponin Ⅴ, and the CAS number of the saponin Ⅴ are
88901-36-4, molecular formula C60H102O29, its chemical structural formula is as follows：
Preferably, the triterpene glucoside is triterpene glucoside VI, and the CAS number of the triterpene glucoside VI are
89590-98-7, molecular formula C66H112O34, its chemical structural formula is as follows：
Preferably, the triterpene glucoside is triterpene glucoside III, and the CAS number of the triterpene glucoside III are
130567-83-8, molecular formula C48H82O19, its chemical structural formula is as follows：
Preferably, the triterpene glucoside is triterpene glucoside II, and the CAS number of the triterpene glucoside II are
88901-38-6, molecular formula C42H72O14, there is following chemical structural formula：
Preferably, the triterpene glucoside is triterpene glucoside IV, and the CAS number of the triterpene glucoside IV are
89590-95-4, molecular formula C54H92O24, there is following chemical structural formula：
Preferably, the radiosensitizer includes the triterpene glucoside and its pharmaceutically acceptable salt and acceptable
Carrier；The acceptable carrier is one or more biocompatible solids or liquid filler or gelatinous mass.They are suitable for people
Use and it is necessary to have enough purity and sufficiently low toxicity.Between " compatibility " referred to herein as each component and each group
Point and the present invention in can mutually admix between triterpene glucoside, and significantly reduce drug effect.The acceptable carrier includes
Cellulose and its derivates, gelatin, talcum, kollag, calcium sulfate, vegetable oil, polyalcohol, emulsifying agent, wetting agent, coloring
One or more in agent, flavor enhancement, stabilizer, antioxidant, preservative, apirogen water.
Preferably, the radiosensitizer is oral liquid, granule, tablet, electuary, capsule and pill, capsule, sustained release
Any one in agent, pill or mouth disintegrant.
The method of application for the tumour radiotherapy sensitizer being prepared using triterpene glucoside is not particularly limited, representational
Method of application includes：Orally, inject, knurl is interior and topical modes.
Solid dosage forms for oral administration includes any one in capsule, tablet, pill, powder or granule.
In these solid dosage forms, reactive compound mixes with least one conventional inert excipients or carrier, such as Dicalcium Phosphate or lemon
Lemon acid sodium, or mixed with following component：(a) filler or solubilizer, for example, lactose, starch, sucrose, mannitol, glucose or silicon
Acid；(b) adhesive, for example, hydroxymethyl cellulose, alginates, gelatin, PVP, sucrose and Arabic gum；
(c) NMF, for example, glycerine；(d) disintegrant, for example, agar, calcium carbonate, farina or tapioca, alginic acid, some
Composition silicate and sodium carbonate；(e) retarding solvent, such as paraffin；(f) absorbsion accelerator, for example, quaternary ammonium compound；(g) soak
Agent, such as cetanol and glycerin monostearate；(h) adsorbent, for example, kaolin；Lubricant, for example, talcum, hard (i)
Resin acid calcium, magnesium stearate, solid polyethylene glycol, lauryl sodium sulfate, or its mixture.
Solid dosage forms such as tablet, sugar-pill, capsule, pill and granule can use coating and shell material to prepare, such as casing and
Other materials well known in the art.They can include opacifying agent, also, reactive compound or compound in this composition
Release can be discharged in certain part in alimentary canal in a delayed fashion.The example of adoptable embedding component is polymeric material
And Wax.If necessary, reactive compound also can be with one or more formation microencapsulation forms in above-mentioned excipient.
Liquid formulation for oral administration includes pharmaceutically acceptable emulsion, solution, suspension, syrup or tincture.
Except active ingredient beyond the region of objective existence, liquid dosage form can include the inert diluent routinely used in this area, such as water or other solvents, increase
Solvent and emulsifying agent, example know, ethanol, ethyl carbonate, isopropanol, 1,3-BDO ethyl acetate, propane diols, dimethylformamide
And oil, the particularly mixture of cottonseed oil, peanut oil, maize germ, olive oil, castor oil and sesame oil or these materials
Deng.
In addition to these inert diluents, the tumour radiotherapy sensitizer can also include auxiliary agent, as wetting agent, emulsifying agent and
Suspending agent, sweetener, flavouring and spices.
Except active ingredient beyond the region of objective existence, suspension can include suspending agent, for example, ethoxylation isooctadecane alcohol, polyoxyethylene
Sorbierite and the mixture of Isosorbide Dinitrate, microcrystalline cellulose, aluminium methoxide and agar or these materials etc..
Tumour radiotherapy sensitizer for parenteral injection can include physiologically acceptable sterile, aqueous or anhydrous solution,
Dispersion liquid, suspension or emulsion, and for being dissolved into the aseptic powdery of sterile Injectable solution or dispersion liquid again.Suitable
Aqueous and nonaqueous carrier, diluent, solvent or excipient include water, ethanol, polyalcohol and its suitable mixture.
The formulation of tumour radiotherapy sensitizer for being locally administered includes ointment, powder, patch, propellant and suction
Agent.Active component aseptically with physiologically acceptable carrier and any preservative, buffer, or if necessary may need
The propellant wanted is mixed together.
Tumour radiotherapy sensitizer of the present invention can be administered alone, or with other pharmaceutically acceptable compounds
Administering drug combinations.
It is that the medicine of safe and effective amount is applicable to treatment during using tumour radiotherapy sensitizer of the present invention
Mammal (such as people), wherein dosage is the effective dosage pharmaceutically thought when applying,
The scope of each dosage, such as the people of 65kg body weight, day dosage be usually 100~400mg.Tool
Body dosage is also contemplated that the factors such as method of administration, patient health situation.
Preferably, the tumour radiotherapy sensitizer is being prepared with the expression and/or downward that can raise P53 tumor suppressor genes
Application in the medicine of the expression of Bcl-2 albumen.
P53 can make apoptosis of tumor cells, so as to prevent histocyte canceration as important antioncogene.While P53 is also
With the function of helping cytogene to repair defect.Bcl-2 is anti-apoptotic proteins, can suppress the procedural apoptosis of cell.Through
Research finds that p53, Bcl-2 gene have equally played important function in radio therapy sensitization, by promoting p53 to express or suppressing
The Bcl-2 effect for reaching radio therapy sensitization.In the present invention, it has been investigated that, it is thin that triterpene glucoside can raise tumour
The expression of P53 tumor suppressor genes or the expression of downward Bcl-2 albumen in born of the same parents.This discovery discloses triterpene glucoside as radiation
Mechanism of action during sensitizer：I.e. triterpene glucoside is possibly through the expression for promoting P53 tumor suppressor genes or suppression Bcl-2 table
Reach to promote the apoptosis of tumour cell, so as to improve radiosensitivity.
Compared with prior art, beneficial effects of the present invention are：
(1) application of the triterpene glucoside in tumour radiotherapy sensitizer is prepared is provided, it is so far there is not yet related
Report.
(2) following nude mice is proposed：Triterpene glucoside may be by promoting tumour cell P53 suppression cancer bases
The expression of cause suppresses Bcl-2 expression to improve radiosensitivity.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the required accompanying drawing used in technology description to be briefly described.
Fig. 1 is the experimental result that triterpene glucoside II regulates and controls P53, Bcl-2 protein expression in Hep-G2 liver cancer cells；
Fig. 2 is different disposal (blank group, pure medicine group, pure radiation group and medicine, radiation joint group) to Hep-G2 liver cancer
The influence of Apoptosis；
Fig. 3 is different disposal (blank group, pure medicine group, pure radiation group and medicine, radiation joint group) to Hep-G2 liver cancer
The influence of cellular morphology；
Fig. 4 is the experimental result that saponin Ⅴ regulates and controls P53, Bcl-2 protein expression in lung cell A549；
Fig. 5 is that different disposal (blank group, pure medicine group, pure radiation group and medicine, radiation joint group) is thin to A549 lung cancer
The influence of born of the same parents' apoptosis；
Fig. 6 is that different disposal (blank group, pure medicine group, pure radiation group and medicine, radiation joint group) is thin to A549 lung cancer
The influence of born of the same parents' form.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition suggested according to normal condition or manufacturer are carried out.Agents useful for same or the unreceipted production firm person of instrument, it is
The conventional products that can be obtained by commercially available purchase.
The triterpene glucoside being previously mentioned in the present invention is prepared with the following method：
1) Momordica grosvenori is crushed, weighed, be 1 by Momordica grosvenori and water quality ratio:6-1:8 ratio adds water, at 80-95 DEG C
At a temperature of stirring extraction 1-3 hour, supernatant is collected by centrifugation, will precipitation repeat stirring extraction 1-4 time, every time centrifuge receipts
Collect supernatant, merge supernatant to obtain extract solution；
2) flocculant is added into above-mentioned extract solution, removes tannin and soluble protein in the extract solution, obtain clear
The clear aqueous solution；
3) the above-mentioned aqueous solution is adsorbed using XAD-16 resins, is then eluted, obtained with 30-50% ethanol
It is enriched with the water-ethanol mixed solution of Momordica grosvenori triterpenoid saponin；
4) above-mentioned water-ethanol mixed solution is concentrated under reduced pressure, and reclaims ethanol, be concentrated into the medicinal extract and title of paste
Weight, the deionized water that 3-6 times of quality is added into the medicinal extract dilutes medicinal extract, to obtain crude product Momordica grosvenori triterpenoid saponin water-soluble
Liquid；
5) decolorization is carried out to the above-mentioned crude product Momordica grosvenori triterpenoid saponin aqueous solution using Diaion PA resins, under collection
Fluid injection, pregnant solution is obtained, recycle semi-preparative liquid chromatography separation that the Momordica grosvenori triterpenoid saponin that purity is more than 98% is made.
On step 1), the stirring 1-3 hour of extraction at a temperature of 80-95 DEG C, 2 hours are preferably extracted；Centrifugation is received
Collect supernatant, precipitation is repeated into stirring extraction 1-4 times, supernatant is collected by centrifugation every time, preferably repeats extraction 1 time, but
It is that the extract solution that the number for repeating to extract more at most obtains is more.
On step 2), flocculant used is chitosan, is existing matured product in the prior art, belongs to organic high
Molecular flocculant.
On step 3), XAD-16 resins are specially Amberlite XAD-16 nonionic macroreticular resins, are generally used for
The absorption of the small molecules such as antibiotic, terpene.
On step 5), Diaion PA resins are specially Mitsubishi Chemical anion exchange resin porous type-Diaion PA
Series, it is mainly used in the decolouring of Momordica grosvenori triterpenoid saponin herein；The chromatographic condition of semi-preparative liquid chromatography：Chromatographic column is anti-phase
C18 posts (it is nonpolar chromatographic column), ultraviolet detection wavelength is 214 ± 2.0nm；Mobile phase is acetonitrile-water, gradient elution 0-
The acetonitriles of 20min 40%, 20-40min 40-60% acetonitriles, the acetonitriles of 40-60min 60%；Flow velocity is 1.0mL/min；Column temperature 25
℃；Sample is collected by different retention times, according to contained glycosyl number is more, retention time is shorter, appearance is faster, is received successively
Collect triterpene glucoside VI (22min), saponin Ⅴ (27min), the triterpene glucoside IV being discussed further below
(35min), triterpene glucoside III (41min), triterpene glucoside II (46min).
Experimental example 1
It is 88901-38-6, molecular formula C that this experimental example, which demonstrates CAS number,42H72O14Triterpene glucoside II it is right
The radiosensitization effect of liver cancer cells.
1st, using Western Blot methods (i.e. WB methods) detect triterpene glucoside II to P53 in liver cancer cells Hep-G2,
Bcl-2 influence
By Hep-G2 cells in various concentrations medicine (0 μm of olL-1, 10 μm of olL-1, 30 μm of olL-1, 60 μm of olL-1)
Cell culture is terminated after thing culture 24h, absorbs nutrient solution afterwards, uses PBS (concentration is 0.01mol L-1, pH 7.4) to wash, adds
Enter the μ L/ holes of lysate containing PMSF 50, put in ice bath environment and crack 30min.Afterwards in 14000r min-1Rotating speed under centrifuge 10min,
Obtain total protein.Protein concentration is surveyed using BCA colorimetric methods, takes 50 μ g total proteins, through 12%SDS polyacrylamide gel electrophoresises point
From rear, electrotransfer to pvdf membrane, 5% skim milk (containing 0.1%Tween 20) closing lh, add ANTIBODY-p53, Bcl-2 and β-
Actin, 4 DEG C of primary antibody are incubated overnight (β-actin compare as applied sample amount)；TBS-T washes film 3 times, each 5min；Add horseradish peroxide
The secondary antibody of compound enzyme (HRP) mark, is incubated at room temperature 1h, washes film 3 times, each 10min with rinsing liquid (TBS-T), adds ECL lucifuges
5min is incubated, the development of fluoroscopic image analyzer, scanning, is analyzed, detection detection P53, Bcl-2, β-actin protein expression levels,
Its result is as shown in Figure 1.
Such as Fig. 1 as can be seen that the expression of the Bcl-2 albumen of the Hep-G2 liver cancer cells handled by triterpene glucoside II is bright
It is aobvious to decline, and the concentration of the expression quantity of Bcl-2 albumen and triterpene glucoside II present it is negatively correlated, and P53 protein expressions it is obvious on
Adjust, and positive correlation is presented in the concentration of expression quantity and triterpene glucoside II.
It can be seen that triterpene glucoside has the function that to raise tumour cell p53 and lowers Bcl-2, therefore, triterpene glucoside II
The potentiality of radiation sensitivity with increase tumour cell, can be prepared tumour radiotherapy sensitizer.
2nd, the influence of blank group, pure medicine group, pure radiation group, joint group (irradiation+medicine) to hepatoma cell apoptosis
Utilize influence of the Flow Cytometry detection triterpene glucoside II to hepatoma cell apoptosis.Collect different grouping culture
Cell, wash cell twice with cold 200 μ L PBS, collect cell；Add 50 μ L combination liquid (Binding Buffer) weight
Outstanding cell, the Annexin V-FITC for adding 2 μ L are mixed, and are added 5 μ L PI and are mixed, and lucifuge, room temperature effect 10min, carry out streaming
Cell instrument detects.Its testing result is as shown in Figure 2.
From figure 2 it can be seen that medicine group, radiation group play the role of substantially to suppress growth of tumour cell；Joint group
Action effect is best, and hepatoma cell apoptosis number substantially increases, and illustrates that triterpene glucoside II has the effect of enhanced sensitivity.
3rd, the influence of blank group, pure medicine group, pure radiation group, joint group (irradiation+medicine) to Morphology of Hepatocellular Carcinoma
Observation irradiation group, the influence of medicine group, joint group (irradiation+medicine) to Morphology of Hepatocellular Carcinoma are dyed using tunel：
Cell is washed with PBS 1 time；Cell is fixed with 4% paraformaldehyde 30 minutes；Washed 1 time with PBS；Addition contains 0.1%Triton
X-100 PBS, ice bath are incubated 2 minutes, rupture of membranes；Washed 1 time with PBS；0.3% hydrogenperoxide steam generator prepared with methanol
(0.3%H2O2In Methanol) in incubation at room temperature 20 minutes, cut into slices endogenous peroxidase with inactivation.Then washed with PBS
Wash 3 times；Add 50 μ l biotin labeling liquid on sample, 37 DEG C are incubated 60 minutes；Washed 1 time with PBS, it is anti-that 0.2ml marks are added dropwise
Terminate liquid is answered, is incubated at room temperature 10 minutes；Washed 3 times with PBS；Add 50 μ l Streptavidin-HRP working solutions, room on sample
Temperature is incubated 30 minutes；Washed 3 times with PBS；0.4ml DAB nitrite ions are added dropwise, are incubated at room temperature 15 minutes；Washed 3 times with PBS；With
Haematoxylin dyeing liquid carries out nuclear targeting.Then washed 3 times with PBS；Direct microscope is observed.Its result such as Fig. 3 institutes
Show.
After tunel dyeing, normal cancer cell does not colour；Occur apoptosis cancer cell by brown, darken, wither
The liver cancer cells died diminish, karyopycnosis.As can be seen from Figure 3：Pure medicine group, pure radiation group, joint group have promotion liver cancer
Hepatoma cell apoptosis phenomenon is the most obvious in the effect of Apoptosis, wherein joint group, and nuclei dyeing is into depth in liver cancer cells
Brown.This result also indicates that triterpene glucoside II has radiosensitizing effect.
4th, colony formation
Colony formation determines cellular radiosensitivity.Hep-G2 cells are diluted to 1 × 104/ mL, 96 orifice plates are added,
Per the μ L of hole 100.It is divided into pure irradiation group, joint group (medicine+irradiation group).Pre-irradiation, to being handled addition sieve in medicine+irradiation group
Chinese fruit saponin(e II (its concentration in joint group is 10 μm of ol/L) acts on 24h.Then, at room temperature, using 6MV-X line singles
Irradiation, dosage are respectively 0Gy, 2Gy, 4Gy, 6Gy, 8Gy, irradiate condition：6MV-X lines, room temperature irradiation, field size 15cm ×
15cm, add the equivalent tissue fillers of 1.5cm.After irradiation, change nutrient solution and continue to discard each hole supernatant after being incubated 2 weeks,
Formaldehyde is fixed, Giemsa dyeing, and meter statistical result, calculates thin containing clone's numbers more than more than 50 cells under inverted microscope
Born of the same parents' survival rate.CNN surviving fraction SF2=(experimental group mean OD value/blank control group mean OD value) × 100%, enhance-ment ratio
SER=radiocontrast groups SF/ (medicine group+radiation group SF).Model SF=1- (1-e are clicked using more targets-D/D0)NIt is quick to calculate radiation
Perceptual related parameter D0、Dq、N、K.Experiment is repeated 3 times, and is averaged, and is carried out curve fitting and is counted with the softwares of prism 5
Calculate related radiobiological parameters.
Liver cancer cells CNN surviving fraction after the various dose radiation exposure of table 1
Influence of the triterpene glucoside II of table 2 for Hep-G2 radiosensitivities
| Different disposal | D0/Gy | SF2/ % | Dq/Gy | N | SER |
| Irradiation group | 2.32 | 58.01 | 1.20 | 1.68 | -- |
| Joint group | 1.68 | 39.23 | 0.55 | 1.38 | 1.48 |
Experimental data shows the D of medication group0、SF2, Dq and N be significantly lower than simple irradiation group.D0As multitarget theory
Important parameter, represent D0Refer to primary emission and kill dosage needed for 63% cell, D0The meaning that diminishes medicine adds cell to ray
Sensitiveness, add D after medicine01.68 are down to from 2.32, triterpene glucoside II adds sensitiveness of the liver cancer cells to ray；SF2
It can directly reflect that triterpene glucoside II on cell sensitive influence, adds SF after medicine239.23% is reduced to, experimental result is said
Bright triterpene glucoside II has sensitization to liver cancer cells.Dq (Dq) reflects the reparation energy of cell sublethal damage
Power, Dq, which diminishes, illustrates that liver cancer cells subdirectory irreducible rings ability reduces.The result of experiment shows that triterpene glucoside II is right
Hep-G2 liver cancer cells have enhancement effect in radiation sensitivity experiment.
Embodiment 2
It is 88901-36-4, molecular formula C that this experimental example, which demonstrates CAS number,60H102O29Saponin Ⅴ pair
The radiosensitization effect of lung carcinoma cell.
1st, influence of the saponin Ⅴ to P53, Bcl-2 in lung carcinoma cell
Utilize influence of the protein immunization experiment Western Blot detection saponin Ⅴs to P53, Bcl-2.
By Hep-G2 cells in various concentrations medicine (0 μm of olL-1, 10 μm of olL-1, 30 μm of olL-1, 60 μm of olL-1)
Cell culture is terminated after thing culture 24h, absorbs nutrient solution afterwards, uses PBS (concentration is 0.01mol L-1, pH 7.4) to wash, adds
Enter the μ L/ holes of lysate containing PMSF 50, put in ice bath environment and crack 30min.Afterwards in 14000r min-1Rotating speed under centrifuge 10min,
Obtain total protein.Protein concentration is surveyed using BCA colorimetric methods, takes 50 μ g total proteins, through 12%SDS polyacrylamide gel electrophoresises point
From rear, electrotransfer to pvdf membrane, 5% skim milk (containing 0.1%Tween 20) closing lh, add antibody P53, Bcl-2 and β-
Actin, 4 DEG C of primary antibody are incubated overnight (β-actin compare as applied sample amount)；TBS-T washes film 3 times, each 5min；Add horseradish peroxide
The secondary antibody of compound enzyme (HRP) mark, is incubated at room temperature 1h, washes film 3 times, each 10min with rinsing liquid (TBS-T), adds ECL lucifuges
5min is incubated, the development of fluoroscopic image analyzer, scanning, is analyzed, detection detection P53, Bcl-2, β-actin protein expression levels,
Its result is as shown in Figure 4.
As shown in figure 4, the expression of the Bcl-2 albumen of the A549 cells handled by saponin Ⅴ is decreased obviously, and
Negative correlation is presented in the expression quantity of Bcl-2 albumen and the concentration of saponin Ⅴ, and P53 protein expressions substantially raise, and expresses
Positive correlation is presented in amount and the concentration of saponin Ⅴ.
In embodiment, saponin Ⅴ has the function that to raise tumour cell P53 and lowers Bcl-2, therefore, Momordica grosvenori
Saponin(e has the potentiality of the radiation sensitivity of increase tumour cell, can be used as preparing tumour radiotherapy sensitizer.2nd, blank group,
The influence of pure medicine group, pure radiation group, joint group (irradiation+medicine) to Increase Apoptosis of Lung Cancer Cells
Utilize influence of the Flow Cytometry detection saponin Ⅴ to Increase Apoptosis of Lung Cancer Cells.Collect different grouping culture
Cell, wash cell twice with cold 200 μ L PBS, collect cell；Add 50 μ L combination liquid (Binding Buffer) weight
Outstanding cell, the Annexin V-FITC for adding 2 μ L are mixed, and are added 5 μ L PI and are mixed, and lucifuge, room temperature effect 10min, carry out streaming
Cell instrument detects.Its testing result is as shown in Figure 5.
From figure 5 it can be seen that medicine group, radiation group play the role of substantially to suppress growth of tumour cell；Joint group
Action effect is best, and hepatoma cell apoptosis number substantially increases, and illustrates that saponin Ⅴ has the effect of enhanced sensitivity.
3rd, the influence of blank group, pure medicine group, pure radiation group, joint group (irradiation+medicine) to pneumonocyte form
After the packet culture of A549 cells, waste liquid is suctioned out, adds 0.5mL fixers per hole, fixed 25min, PBS wash 2 times, often
It is secondary to wash 3min, add the dye liquors of Hoechst 33258, room temperature lucifuge dyeing 20min.Become using fluorescence microscope cellular morphology
Change.Its testing result is as shown in Figure 6.
From fig. 6 it can be seen that blank group nucleus is complete, uniform coloring, fluorescence does not occur Apoptosis into disperse shape
Phenomenon；In medicine group, radiation group, there is part and be in granular form fluorescence shape apoptotic cell, the effect of radiation group is better than medicine group；
Occur a large amount of shrinkages, the cell of apoptosis in joint group, illustrate that joint group has strong inhibition effect to lung carcinoma cell.
4th, colony formation
Colony formation determines cellular radiosensitivity.A549 cells are diluted to 1 × 104/ mL, 96 orifice plates are added, often
The μ L of hole 100.It is divided into pure irradiation group, joint group (medicine+irradiation group).Pre-irradiation, to being handled addition arhat in medicine+irradiation group
Fruit saponin(e V (its concentration in joint group is 10 μm of ol/L) acts on 24h.Then, at room temperature, shone using 6MV-X lines single
Penetrate, dosage is respectively 0Gy, 2Gy, 4Gy, 6Gy, 8Gy, irradiates condition：6MV-X lines, room temperature irradiation, field size 15cm ×
15cm, add the equivalent tissue fillers of 1.5cm.After irradiation, change nutrient solution and continue to discard each hole supernatant after being incubated 2 weeks,
Formaldehyde is fixed, Giemsa dyeing, and meter statistical result, calculates thin containing clone's numbers more than more than 50 cells under inverted microscope
Born of the same parents' survival rate.CNN surviving fraction SF2=(experimental group mean OD value/blank control group mean OD value) × 100%, enhance-ment ratio
SER=radiocontrast groups SF/ (medicine group+radiation group SF).Model SF=1- (1-e are clicked using more targets-D/D0)NIt is quick to calculate radiation
Perceptual related parameter D0、Dq、N、K.Experiment is repeated 3 times, and is averaged, and is carried out curve fitting and is counted with the softwares of prism 5
Calculate related radiobiological parameters.
Lung carcinoma cell CNN surviving fraction after the various dose radiation exposure of table 3
Influence of the saponin Ⅴ of table 4 for A549 radiosensitivities
| Different disposal | D0/Gy | SF2/% | Dq/Gy | N | SER |
| Irradiation group | 2.33 | 64.01 | 1.60 | 1.99 | -- |
| Joint group | 1.91 | 47.23 | 0.79 | 1.51 | 1.33 |
Experimental data shows the D of medication group0、SF2, Dq and N be significantly lower than simple irradiation group.D0As multitarget theory
Important parameter, represent D0Refer to primary emission and kill dosage needed for 63% cell, D0The meaning that diminishes medicine adds cell to ray
Sensitiveness, add D after medicine01.91 are down to from 2.33, saponin Ⅴ adds sensitiveness of the liver cancer cells to ray；SF2
It can directly reflect that saponin Ⅴ on cell sensitive influence, adds SF after medicine247.23% is reduced to, experimental result is said
Bright saponin Ⅴ has sensitization to liver cancer cells.The reparation of the sublethal damage of Dq (Dq) reflection cells
Ability, the explanation liver cancer cells subdirectory irreducible rings ability that diminishes reduce.The result of experiment shows saponin Ⅴ to lung
JEG-3 A549 has enhancement effect in radiation sensitivity experiment.
Embodiment 3
This experimental example demonstrates CAS number89590-98-7, molecular formula C66H112O34Triterpene glucoside VI to palace
The radiosensitization effect of neck cancer Hela cells.
Clone forming experiment
Colony formation determines cellular radiosensitivity.Hela cells are diluted to 1 × 104/ mL, 96 orifice plates are added, often
The μ L of hole 100.It is divided into pure irradiation group, joint group (medicine+irradiation group).Pre-irradiation, to being handled addition arhat in medicine+irradiation group
Fruit saponin(e VI (its concentration in joint group is 10 μm of ol/L) acts on 24h.Then, at room temperature, shone using 6MV-X lines single
Penetrate, dosage is respectively 0Gy, 2Gy, 4Gy, 6Gy, 8Gy, irradiates condition：6MV-X lines, room temperature irradiation, field size 15cm ×
15cm, add the equivalent tissue fillers of 1.5cm.After irradiation, change nutrient solution and continue to discard each hole supernatant after being incubated 2 weeks,
Formaldehyde is fixed, Giemsa dyeing, and meter statistical result, calculates thin containing clone's numbers more than more than 50 cells under inverted microscope
Born of the same parents' survival rate.CNN surviving fraction SF2=(experimental group mean OD value/blank control group mean OD value) × 100%, enhance-ment ratio
SER=radiocontrast groups SF/ (medicine group+radiation group SF).Model SF=1- (1-e are clicked using more targets-D/D0)NIt is quick to calculate radiation
Perceptual related parameter D0、Dq、N、K.Experiment is repeated 3 times, and is averaged, and is carried out curve fitting and is counted with the softwares of prism 5
Calculate related radiobiological parameters.
Hela cell survival fractions after the various dose radiation exposure of table 5
Influence of the triterpene glucoside VI of table 6 for Hela radiosensitivities
| Different disposal | D0/Gy | SF2/% | Dq/Gy | N | SER |
| Irradiation group | 4.01 | 82 | 3.31 | 2.29 | -- |
| Joint group | 3.57 | 75 | 2.23 | 1.87 | 1.09 |
Experimental data shows the D of medication group0、SF2, Dq and N be below simple irradiation group.D0It is important as multitarget theory
Parameter, represent D0Refer to primary emission and kill dosage needed for 63% cell, D0The meaning that diminishes medicine adds sensitivity of the cell to ray
Property, add D after medicine03.57 are down to from 4.01, triterpene glucoside VI adds sensitiveness of the Hela cells to ray；SF2Can be straight
The reversed triterpene glucoside VI that reflects adds SF after medicine to cell sensitive influence2Reduce, experimental result illustrates triterpene glucoside VI
There is sensitization to cervical cancer cell.The repair ability of the sublethal damage of Dq (Dq) reflection cells, becomes novel
Bright liver cancer cells subdirectory irreducible rings ability reduces.The result of experiment shows triterpene glucoside VI to cervical carcinoma JEG-3
Hela has enhancement effect in radiation sensitivity experiment.
Application examples 1
By triterpene glucoside IV to the oral tablet for preparing tumour radiotherapy effect of enhanced sensitivity.
The 500g of triterpene glucoside IV is taken, adds after appropriate dextrin is well mixed and adds the obtained softwood of distilled water, with system
Grain mechanism obtains dry particle, and a diameter of 1cm semi-finished product tablet is made using tablet press machine after sterilizing, after be put into ultraviolet rays environment
15 minutes sterilizing, thus obtaining the product oral tablet product of middle irradiation, the content of every saponin(e of tablet IV is 60mg.
Selected condition：Cattell scoring >=70；Proved by pathology is non-small cell lung cancer；Clinical stages：II~III phase；Without obvious
The heart, lung, Liver and kidney function are abnormal；80 Patients with Non-small-cell Lung that will be admitted to hospital are randomly divided into two groups, enhanced sensitivity group 39：In radiotherapy
Start to take oral tablet within 1st day, each one (the every middle 60mg of saponin(e IV), 3 times a day, until radiotherapy terminates.Irradiation side
Method uses conventional fractionation fluconazole ear drops, is irradiated using 15MVX lines, 66~70Gy/33 of accumulated dose~35f, completes within 6~7 weeks.
Control group 41：Radiation alone, method is the same as enhanced sensitivity group.
The enhanced sensitivity group of table 7 is compared with the recent radiotherapeutic effect of control group.
1) P compared with control group<0.05
Note：Bracket is percentage
As a result shown in table 5, efficacy assessment standard, including complete incidence graph (CR), part alleviate (PR), objective effectively (CR+
PR), display enhanced sensitivity group significant effect is better than control group, and saponin(e IV has certain effect of enhanced sensitivity.
Although illustrate and describing the present invention with specific embodiment, but will be appreciated that without departing substantially from the present invention's
Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (9)
1. the application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared, it is characterised in that
The triterpene glucoside has following structure formula：
In formula, R and R1For glucose residue, the chemical constitution of the glucose residue includes
In any one.
2. triterpene glucoside according to claim 1 or its pharmaceutically acceptable salt are in tumour radiotherapy sensitizer is prepared
Application, it is characterised in that the triterpene glucoside is saponin Ⅴ, and the CAS number of the saponin Ⅴ are
88901-36-4, molecular formula C60H102O29。
3. triterpene glucoside according to claim 1 or its pharmaceutically acceptable salt are in tumour radiotherapy sensitizer is prepared
Application, it is characterised in that the triterpene glucoside is triterpene glucoside VI, and the CAS number of the triterpene glucoside VI are
89590-98-7, molecular formula C66H112O34。
4. triterpene glucoside according to claim 1 or its pharmaceutically acceptable salt are in tumour radiotherapy sensitizer is prepared
Application, it is characterised in that the triterpene glucoside is triterpene glucoside III, and the CAS number of the triterpene glucoside III are
130567-83-8, molecular formula C48H82O19。
5. triterpene glucoside according to claim 1 or its pharmaceutically acceptable salt are in tumour radiotherapy sensitizer is prepared
Application, it is characterised in that the triterpene glucoside is triterpene glucoside II, and the CAS number of the triterpene glucoside II are
88901-38-6, molecular formula C42H72O14。
6. triterpene glucoside according to claim 1 or its pharmaceutically acceptable salt are in tumour radiotherapy sensitizer is prepared
Application, it is characterised in that the triterpene glucoside is triterpene glucoside IV, and the CAS number of the triterpene glucoside IV are
89590-95-4, molecular formula C54H92O24。
7. the triterpene glucoside or its pharmaceutically acceptable salt according to claim any one of 1-6 are preparing tumour radiotherapy
Application in sensitizer, it is characterised in that the radiosensitizer includes the triterpene glucoside and acceptable carrier；
The acceptable carrier is one or more biocompatible solids or liquid filler or gelatinous mass, including cellulose and its
Derivative, gelatin, kollag, calcium sulfate, vegetable oil, polyalcohol, emulsifying agent, wetting agent, colouring agent, flavor enhancement, stably
One or more in agent, antioxidant, preservative, apirogen water.
8. triterpene glucoside according to claim 7 or its pharmaceutically acceptable salt are in tumour radiotherapy sensitizer is prepared
Application, it is characterised in that the radiosensitizer is oral liquid, granule, tablet, electuary, capsule and pill, capsule, slow
Release any one in agent, pill or mouth disintegrant.
9. triterpene glucoside according to claim 1 or its pharmaceutically acceptable salt are in tumour radiotherapy sensitizer is prepared
Application, it is characterised in that the tumour radiotherapy sensitizer prepare with can raise P53 tumor suppressor genes expression and/or
Lower the application in the medicine of the expression of Bcl-2 albumen.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510498383.4A CN105232565B (en) | 2015-08-13 | 2015-08-13 | The application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared |
| GB1610189.1A GB2544360B (en) | 2015-08-13 | 2016-06-10 | Use of mogrosides or the pharmaceutically acceptable salts thereof in the manufacture of a tumor radiation sensitizing agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510498383.4A CN105232565B (en) | 2015-08-13 | 2015-08-13 | The application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105232565A CN105232565A (en) | 2016-01-13 |
| CN105232565B true CN105232565B (en) | 2017-12-08 |
Family
ID=55030564
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510498383.4A Active CN105232565B (en) | 2015-08-13 | 2015-08-13 | The application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN105232565B (en) |
| GB (1) | GB2544360B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105801658B (en) * | 2014-12-30 | 2019-03-19 | 北京农学院 | The preparation of sweet tea glucoside and the like and application as STAT3, ERK signal path target site drug in antitumor |
| CN105784882A (en) * | 2016-04-15 | 2016-07-20 | 广西壮族自治区梧州食品药品检验所 | Method for ASE extraction of triterpene glucoside V in siraitia grosvenorii |
| CN109602757B (en) * | 2019-01-11 | 2020-11-17 | 桂林医学院附属医院 | Application of mogroside IIE in preparation of trypsin inhibitor |
| CN110628703A (en) * | 2019-10-30 | 2019-12-31 | 复旦大学附属金山医院 | Radioactive lung injury in-vitro cell model and construction method and application thereof |
| CN114699394B (en) * | 2022-05-24 | 2022-09-13 | 中山大学附属第一医院 | Pharmaceutical composition for radiotherapy sensitization of liver cancer and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526073A (en) * | 2012-01-06 | 2012-07-04 | 复旦大学 | Application of mogrol H9 for preparing antitumor drugs |
| CN102860498A (en) * | 2011-07-04 | 2013-01-09 | 吉林博大农林生物科技有限公司 | Method for processing Chinese forest frog radiation resistant oral liquid |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003277270A (en) * | 2002-03-20 | 2003-10-02 | Univ Nihon | Carcinogensis preventing agent |
| US9603848B2 (en) * | 2007-06-08 | 2017-03-28 | Senomyx, Inc. | Modulation of chemosensory receptors and ligands associated therewith |
| KR20140133016A (en) * | 2013-05-09 | 2014-11-19 | (주)화평디엔에프 | Manufacturing methods of Siraitia grosvenorii extracts, and composition comprising the same |
| WO2016001907A1 (en) * | 2014-07-02 | 2016-01-07 | Prendergast Patrick T | Mogroside iv and mogroside v as agonist/stimulator/un-blocking agent for toll-like receptor 4 and adjuvant for use in human/animal vaccine and to stimulate immunity against disease agents. |
-
2015
- 2015-08-13 CN CN201510498383.4A patent/CN105232565B/en active Active
-
2016
- 2016-06-10 GB GB1610189.1A patent/GB2544360B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102860498A (en) * | 2011-07-04 | 2013-01-09 | 吉林博大农林生物科技有限公司 | Method for processing Chinese forest frog radiation resistant oral liquid |
| CN102526073A (en) * | 2012-01-06 | 2012-07-04 | 复旦大学 | Application of mogrol H9 for preparing antitumor drugs |
Non-Patent Citations (1)
| Title |
|---|
| 罗汉果皂苷生物活性研究进展;曹建平 等;《食品工业科技》;20141231;第35卷(第24期);第387页左栏第2段-右栏第1段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| GB2544360A (en) | 2017-05-17 |
| CN105232565A (en) | 2016-01-13 |
| GB2544360B (en) | 2017-11-01 |
| GB201610189D0 (en) | 2016-07-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105232565B (en) | The application of triterpene glucoside or its pharmaceutically acceptable salt in tumour radiotherapy sensitizer is prepared | |
| Gupta et al. | Neuroprotective potential of ellagic acid: a critical review | |
| Hong et al. | Novel ginsenoside-based multifunctional liposomal delivery system for combination therapy of gastric cancer | |
| Yu et al. | Zinc phthalocyanine encapsulated in polymer micelles as a potent photosensitizer for the photodynamic therapy of osteosarcoma | |
| US20130259893A1 (en) | Food recipes for nourishing, maintaining and cultivating a variety of stem cells and a method for manufacturing the same | |
| Yuan et al. | In vitro and in vivo inhibitory effects of a Pleurotus eryngii protein on colon cancer cells | |
| EP2908833B1 (en) | Formulations comprising herbal extracts | |
| JP6656316B2 (en) | How to use cucumbers, how to use cucumbers extract and how to use drug mixtures | |
| CN106139148A (en) | A kind of tumor chemotherapeutic medicinal preparation combination | |
| Yapasert et al. | Anticancer effects of a traditional Thai herbal recipe Benja Amarit extracts against human hepatocellular carcinoma and colon cancer cell by targeting apoptosis pathways | |
| Ma et al. | The ethyl acetate extract of Gynura formosana Kitam. leaves inhibited cervical cancer cell proliferation via induction of autophagy | |
| Ou et al. | Autophagy is involved in Ficus carica fruit extract-induced anti-tumor effects on pancreatic cancer | |
| WO2015192758A1 (en) | Anti-tumor pharmaceutical application of pentacyclic triterpene saponin compounds of szechuan melandium root | |
| WO2017173611A1 (en) | Pharmaceutical product for treating tumor and combined immune defect, and preparation and application thereof | |
| CN1994352B (en) | Use of total glucosides extract of cocklebur fruit in preparation of product for resisting inflammatory reaction | |
| US20050147701A1 (en) | Anti-prostate cancer composition and therapeutic uses therefor | |
| TW200400828A (en) | A composition for cancer therapy | |
| KR102360622B1 (en) | Composition comprising extracts of Inonotus obliquus for the Prevention, Treatment or Improving of Radioresistant Cancer | |
| Gupta | In-Vitro anti-inflammatory activity of S. xanthocarpum and A. officinarum herb by Human red blood cell membrane stabilization method | |
| CN104814959B (en) | Magnetic sodium cantharidinate vitamin B6 compound preparation and preparation method thereof | |
| CN104367612B (en) | A kind of application of dog ant grass extract | |
| CN100464760C (en) | Use of cocklebur fruit total glucoside extract in preparing anti inflammation reaction product | |
| KR101021975B1 (en) | A composition for enhancing the radiotherapy of cancer | |
| Khosravi et al. | Pharmacognosy and pharmacology of Calotropis gigantea for discovery of anticancer therapeutics | |
| JP2021109836A (en) | Composition for reducing side effect associated with cancer chemotherapy |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |