CN107441110A - A kind of Exocarpium Citri Grandis extract and its application in cancer therapy drug is prepared - Google Patents
A kind of Exocarpium Citri Grandis extract and its application in cancer therapy drug is prepared Download PDFInfo
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- exocarpium citri
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- 239000000284 extract Substances 0.000 title claims abstract description 66
- 239000003814 drug Substances 0.000 title claims abstract description 24
- 229940079593 drug Drugs 0.000 title claims abstract description 18
- 238000011275 oncology therapy Methods 0.000 title claims abstract description 12
- IYRMWMYZSQPJKC-UHFFFAOYSA-N kaempferol Chemical compound C1=CC(O)=CC=C1C1=C(O)C(=O)C2=C(O)C=C(O)C=C2O1 IYRMWMYZSQPJKC-UHFFFAOYSA-N 0.000 claims abstract description 42
- 239000000203 mixture Substances 0.000 claims abstract description 30
- OBIOZWXPDBWYHB-UHFFFAOYSA-N Nobiletin Natural products C1=CC(OC)=CC=C1C1=C(OC)C(=O)C2=C(OC)C(OC)=C(OC)C(OC)=C2O1 OBIOZWXPDBWYHB-UHFFFAOYSA-N 0.000 claims abstract description 22
- MRIAQLRQZPPODS-UHFFFAOYSA-N nobiletin Chemical compound C1=C(OC)C(OC)=CC=C1C1=CC(=O)C2=C(OC)C(OC)=C(OC)C(OC)=C2O1 MRIAQLRQZPPODS-UHFFFAOYSA-N 0.000 claims abstract description 22
- UBSCDKPKWHYZNX-UHFFFAOYSA-N Demethoxycapillarisin Natural products C1=CC(O)=CC=C1OC1=CC(=O)C2=C(O)C=C(O)C=C2O1 UBSCDKPKWHYZNX-UHFFFAOYSA-N 0.000 claims abstract description 21
- RPMNUQRUHXIGHK-PYXJVEIZSA-N apigenin 7-O-neohesperidoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C=C(C=3C=CC(O)=CC=3)OC2=C1 RPMNUQRUHXIGHK-PYXJVEIZSA-N 0.000 claims abstract description 21
- 229930034861 apigenin-7-O-neohesperidoside Natural products 0.000 claims abstract description 21
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 claims abstract description 21
- 235000008777 kaempferol Nutrition 0.000 claims abstract description 21
- UXOUKMQIEVGVLY-UHFFFAOYSA-N morin Natural products OC1=CC(O)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 UXOUKMQIEVGVLY-UHFFFAOYSA-N 0.000 claims abstract description 21
- RPMNUQRUHXIGHK-SBDOOABHSA-N rhoifolin Natural products O([C@@H]1[C@@H](O)[C@H](O)[C@H](CO)O[C@H]1Oc1cc(O)c2C(=O)C=C(c3ccc(O)cc3)Oc2c1)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](C)O1 RPMNUQRUHXIGHK-SBDOOABHSA-N 0.000 claims abstract description 21
- 201000007270 liver cancer Diseases 0.000 claims abstract description 16
- 208000014018 liver neoplasm Diseases 0.000 claims abstract description 16
- 239000004480 active ingredient Substances 0.000 claims abstract description 7
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 30
- 239000000741 silica gel Substances 0.000 claims description 29
- 229910002027 silica gel Inorganic materials 0.000 claims description 29
- 229960001866 silicon dioxide Drugs 0.000 claims description 29
- 239000002904 solvent Substances 0.000 claims description 29
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 27
- BQFCCCIRTOLPEF-UHFFFAOYSA-N chembl1976978 Chemical compound CC1=CC=CC=C1N=NC1=C(O)C=CC2=CC=CC=C12 BQFCCCIRTOLPEF-UHFFFAOYSA-N 0.000 claims description 26
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- 239000012046 mixed solvent Substances 0.000 claims description 21
- 238000000605 extraction Methods 0.000 claims description 18
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 16
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 14
- 238000010828 elution Methods 0.000 claims description 14
- 239000003208 petroleum Substances 0.000 claims description 13
- 239000000470 constituent Substances 0.000 claims description 12
- 238000010898 silica gel chromatography Methods 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- 239000000469 ethanolic extract Substances 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 10
- 239000000725 suspension Substances 0.000 claims description 9
- 230000001093 anti-cancer Effects 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 7
- 239000012535 impurity Substances 0.000 claims description 7
- 238000010992 reflux Methods 0.000 claims description 7
- 239000012259 ether extract Substances 0.000 claims description 6
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000003560 cancer drug Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims description 2
- 244000044283 Toxicodendron succedaneum Species 0.000 claims 1
- 239000012141 concentrate Substances 0.000 claims 1
- QGBSISYHAICWAH-UHFFFAOYSA-N dicyandiamide Chemical compound NC(N)=NC#N QGBSISYHAICWAH-UHFFFAOYSA-N 0.000 claims 1
- XYIBRDXRRQCHLP-UHFFFAOYSA-N ethyl acetoacetate Chemical compound CCOC(=O)CC(C)=O XYIBRDXRRQCHLP-UHFFFAOYSA-N 0.000 claims 1
- 229930182470 glycoside Natural products 0.000 claims 1
- 150000002338 glycosides Chemical class 0.000 claims 1
- 239000004575 stone Substances 0.000 claims 1
- 201000011510 cancer Diseases 0.000 description 6
- 238000012360 testing method Methods 0.000 description 5
- 244000276331 Citrus maxima Species 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 235000001759 Citrus maxima Nutrition 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 239000012980 RPMI-1640 medium Substances 0.000 description 3
- 230000001644 anti-hepatocarcinoma Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000003810 ethyl acetate extraction Methods 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 206010011224 Cough Diseases 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 229910052710 silicon Inorganic materials 0.000 description 2
- 239000010703 silicon Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 1
- 206010073069 Hepatic cancer Diseases 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 206010036590 Premature baby Diseases 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000007761 synergistic anti-cancer Effects 0.000 description 1
- 210000001519 tissue 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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- 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
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- 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)
- A61K36/75—Rutaceae (Rue family)
- A61K36/752—Citrus, e.g. lime, orange or lemon
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to field of pharmaceutical chemistry technology, specifically discloses a kind of Exocarpium Citri Grandis extract and its application in cancer therapy drug is prepared.The extract includes following active ingredient：Nobiletin, Kaempferol and Rhoifolin, the percentage by weight of each composition are：Nobiletin 30~40%；Kaempferol 20~30%；Rhoifolin 20~30%.Exocarpium Citri Grandis extract active ingredient and content of the present invention are clear and definite, and anti-liver cancer efficacy is notable.
Description
Technical field
The present invention relates to field of pharmaceutical chemistry technology, and in particular to a kind of Exocarpium Citri Grandis extract and its is preparing cancer therapy drug
In application.
Background technology
Exocarpium Citri Grandis, i.e. Huajuhong, source this product are the prematurity or maturescent dry of rutaceae Citrus grandis or shaddock
Dry outside rind.There is preventing phlegm from forming and stopping coughing, treatment cough due to wind-cold evil.
Liver cancer is liver malignancy, can be divided into primary and the major class of Secondary cases two.Characters of Primary Malignant Tumors of Liver originates from
In the epithelium or mesenchymal tissue of liver, the former is referred to as primary carcinoma of liver, is that China is occurred frequently, very harmful malignant tumour.
It is more using pummelo pee medicinal material as wherein medicinal material carrys out treating cancer simply in existing anticancer report, have no exclusive useization
Report of the Exocarpium Citri Rubrum active component as cancer therapy drug.If number of patent application is 201310669278.3 a kind of to treat in lung cancer
The Chinese medicine for being used to treat lung cancer of drug composition and Application No. 201510164588.9, wherein using pummelo pee medicinal material as
Wherein medicinal material carrys out treating cancer simply.
Therefore, based on Exocarpium Citri Grandis, studying a kind of anticancer effect, good cancer therapy drug has wide market prospects.
The content of the invention
The technical problems to be solved by the invention are, in order to overcome in the prior art to anticancer component in Exocarpium Citri Grandis not
Foot, there is provided a kind of Exocarpium Citri Grandis extract, described Exocarpium Citri Grandis extract have the anticancer effect of highly significant.
Above-mentioned technical problem to be solved by this invention, is achieved by the following technical programs：
A kind of Exocarpium Citri Grandis extract, include following active ingredient：Nobiletin, Kaempferol and Rhoifolin.
Preferably, described Exocarpium Citri Grandis extract, the active ingredient of following percentage by weight is included：Nobiletin 30~
40%；Kaempferol 20~30%；Rhoifolin 20~30%.
The preparation method of above-mentioned Exocarpium Citri Grandis extract, is comprised the following steps：
(1) Exocarpium Citri Grandis is taken, with ethanol heating and refluxing extraction, concentrated extracting solution obtains Exocarpium Citri Grandis ethanol extract；
(2) Exocarpium Citri Grandis ethanol extract is dissolved in water into suspension, first with petroleum ether extraction, discards petroleum ether extraction
Thing, then be extracted with ethyl acetate, concentration acetic acid ethyl ester extract obtains Exocarpium Citri Grandis extract；
(3) Exocarpium Citri Grandis extract is produced into Exocarpium Citri Grandis extract through silica gel column chromatography is active constituent-enriched.
Preferably, the ethanol described in step (1) is the ethanol water that volume fraction is 80~100%；Described second
The dosage of alcohol and the amount ratio of Exocarpium Citri Grandis are 8~15mL:1g；The time of described heating and refluxing extraction is 1~3h.
Preferably, petroleum ether extraction is discarded first with the petroleum ether extraction isometric with suspension 1~3 time in step (2)
Thing, then extracted 1~3 time with the isometric ethyl acetate of suspension, concentration acetic acid ethyl ester extract obtains Exocarpium Citri Grandis extract.
Preferably, the active constituent-enriched specific method of silica gel column chromatography is in step (3)：By silicon on Exocarpium Citri Grandis extract
Glue post, it is first 90 with volume ratio:10~87:The mixed solvent system of 13 solvent orange 2 A and solvent B composition carries out elution removal of impurities；So
It is 82 to use volume ratio again afterwards:18~80:The mixed solvent system of 20 solvent orange 2 A and solvent B composition is eluted and collects elution
Position, produce Exocarpium Citri Grandis active component；Described solvent orange 2 A is chloroform, and solvent B is methanol；
Volume ratio is 90:10~87:The dosage of the mixed solvent system of 13 solvent orange 2 A and solvent B composition is silicagel column post
3~5 times of volume；Volume ratio is 82:18~80:The dosage of the mixed solvent system of 20 solvent orange 2 A and solvent B composition is silica gel
5~8 times of post column volume.
Preferably, the active constituent-enriched specific method of silica gel column chromatography is in step (3)：By silicon on Exocarpium Citri Grandis extract
Glue post, it is first 87 with volume ratio:13 solvent orange 2 A：The mixed solvent system of solvent B compositions carries out elution removal of impurities；Then body is used again
Product is than being 82:18 solvent orange 2 A：The mixed solvent system of solvent B compositions is eluted and collects elution position, and producing Exocarpium Citri Grandis has
Imitate position；Described solvent orange 2 A is chloroform, and solvent B is methanol；
Volume ratio is 87:The dosage of the mixed solvent system of 13 solvent orange 2 A and solvent B composition is the 3 of silicagel column column volume
Times；Volume ratio is 82:The dosage of the mixed solvent system of 18 solvent orange 2 A and solvent B composition is 8 times of silicagel column column volume.
The silicagel column column volume refers to after filling post with silica gel, from post bottom to the volume of silica gel deposition surface.
Preferably, the weight of used silica gel is 20~40 times of Exocarpium Citri Grandis extract dry weight in silicagel column.
Application of the above-mentioned Exocarpium Citri Grandis extract in cancer therapy drug is prepared；Described cancer therapy drug is medicines resistant to liver cancer.
The present invention also provides a kind of anticancer pharmaceutical composition from Exocarpium Citri Grandis, and it includes having for following percentage by weight
Imitate composition：Nobiletin, Kaempferol, the weight ratio of Rhoifolin are 3~4：2~3：2~3；Described cancer therapy drug is anti-liver
Cancer drug.
Beneficial effect：The present invention develops a kind of brand-new Exocarpium Citri Grandis extract, and described extract has highly significant
Antihepatocarcinoma effect, overcome prior art to Exocarpium Citri Grandis anticancer position research deficiency；In addition, described Exocarpium Citri Grandis extract
Active ingredient and content are clear and definite, when using the Exocarpium Citri Grandis extract as medicines resistant to liver cancer in use, being advantageous to the quality of medicine
Control and drug safety.
Embodiment
The present invention is explained further below in conjunction with specific embodiment, but embodiment does not do any type of limit to the present invention
It is fixed.
The preparation of the Exocarpium Citri Grandis extract of embodiment 1
(1) Exocarpium Citri Grandis is taken, with the ethanol water heating and refluxing extraction 1.5h that volume fraction is 95%, concentrated extracting solution obtains
Exocarpium Citri Grandis ethanol extract；
(2) Exocarpium Citri Grandis ethanol extract is dissolved in water into suspension, first with isometric petroleum ether extraction, repeats to extract
3 times, petroleum ether extract is discarded, then is extracted with isometric ethyl acetate, repeats extraction 3 times, merges concentration ethyl acetate extraction
Thing is taken to obtain Exocarpium Citri Grandis extract；
(3) Exocarpium Citri Grandis extract is produced into Exocarpium Citri Grandis extract through silica gel column chromatography is active constituent-enriched；
The active constituent-enriched method of the silica gel column chromatography is：By (the institute in silicagel column of silicagel column on Exocarpium Citri Grandis extract
With 30 times that the weight of silica gel is Exocarpium Citri Grandis extract dry weight, the specification of silica gel is 200 mesh), it is first 87 with volume ratio:13 it is molten
Agent A：The mixed solvent system (dosage is 3 times of column volumes) of solvent B compositions carries out elution removal of impurities；Then it is 82 to use volume ratio again:
18 solvent orange 2 A：The mixed solvent system (dosage is 8 times of column volumes) of solvent B compositions is eluted and collects elution position, is produced
Exocarpium Citri Grandis active component；Described solvent orange 2 A is chloroform, and solvent B is methanol.
After testing, Nobiletin of the percentage by weight for 37%, 28% Kaempferol are contained in the Exocarpium Citri Grandis extract
And 26% Rhoifolin.
The content assaying method of mentioned component is (similarly hereinafter)：Using liquid chromatographic detection, with Nobiletin, Kaempferol and
Rhoifolin is measured as standard items using external standard method.
Described liquid phase chromatogram condition is：With Agilent Zorbax SB C18(column length 250mm, internal diameter 4.6mm, particle diameter 5
μm) it is chromatographic column；Using acetonitrile as mobile phase A, using 0.05% acetic acid solution as Mobile phase B, Detection wavelength 330nm；Column temperature
25 DEG C, flow velocity 1.0mL/min；Mobile phase A：Mobile phase B=15:85.The wherein retention time of Rhoifolin is 6.3min, mountain
How the retention time of phenol is 8.8min, and the retention time of Nobiletin is 14.4min.
The preparation of the Exocarpium Citri Grandis extract of embodiment 2
(1) Exocarpium Citri Grandis is taken, with the ethanol water heating and refluxing extraction 1.5h that volume fraction is 95%, concentrated extracting solution obtains
Exocarpium Citri Grandis ethanol extract；
(2) Exocarpium Citri Grandis ethanol extract is dissolved in water into suspension, first with isometric petroleum ether extraction, repeats to extract
3 times, petroleum ether extract is discarded, then is extracted with isometric ethyl acetate, repeats extraction 3 times, merges concentration ethyl acetate extraction
Thing is taken to obtain Exocarpium Citri Grandis extract；
(3) Exocarpium Citri Grandis extract is produced into Exocarpium Citri Grandis extract through silica gel column chromatography is active constituent-enriched；
The active constituent-enriched method of the silica gel column chromatography is：By (the institute in silicagel column of silicagel column on Exocarpium Citri Grandis extract
With 30 times that the weight of silica gel is Exocarpium Citri Grandis extract dry weight, the specification of silica gel is 200 mesh), it is first 90 with volume ratio:10 it is molten
Agent A：The mixed solvent system (dosage is 3 times of column volumes) of solvent B compositions carries out elution removal of impurities；Then it is 82 to use volume ratio again:
18 solvent orange 2 A：The mixed solvent system (dosage is 8 times of column volumes) of solvent B compositions is eluted and collects elution position, is produced
Exocarpium Citri Grandis active component；Described solvent orange 2 A is chloroform, and solvent B is methanol.
After testing, Nobiletin of the percentage by weight for 35%, 24% Kaempferol are contained in the Exocarpium Citri Grandis extract
And 25% Rhoifolin.
The preparation of the Exocarpium Citri Grandis extract of embodiment 3
(1) Exocarpium Citri Grandis is taken, with the ethanol water heating and refluxing extraction 1.5h that volume fraction is 95%, concentrated extracting solution obtains
Exocarpium Citri Grandis ethanol extract；
(2) Exocarpium Citri Grandis ethanol extract is dissolved in water into suspension, first with isometric petroleum ether extraction, repeats to extract
3 times, petroleum ether extract is discarded, then is extracted with isometric ethyl acetate, repeats extraction 3 times, merges concentration ethyl acetate extraction
Thing is taken to obtain Exocarpium Citri Grandis extract；
(3) Exocarpium Citri Grandis extract is produced into Exocarpium Citri Grandis extract through silica gel column chromatography is active constituent-enriched；
The active constituent-enriched method of the silica gel column chromatography is：By (the institute in silicagel column of silicagel column on Exocarpium Citri Grandis extract
With 30 times that the weight of silica gel is Exocarpium Citri Grandis extract dry weight, the specification of silica gel is 200 mesh), it is first 90 with volume ratio:10 it is molten
Agent A：The mixed solvent system (dosage is 3 times of column volumes) of solvent B compositions carries out elution removal of impurities；Then it is 80 to use volume ratio again:
20 solvent orange 2 A：The mixed solvent system (dosage is 8 times of column volumes) of solvent B compositions is eluted and collects elution position, is produced
Exocarpium Citri Grandis active component；Described solvent orange 2 A is chloroform, and solvent B is methanol.
After testing, Nobiletin of the percentage by weight for 32%, 21% Kaempferol are contained in the Exocarpium Citri Grandis extract
And 22% Rhoifolin.
The extract active anticancer of embodiment 4 is tested
Take human liver cancer cell BEL-7404, be placed in culture medium (culture mediums of RPMI 1640) and cultivate, cancer cell with 10 ×
103Added per hole in 96 orifice plates, contain 5%CO at 37 DEG C2Incubator in cultivate 24 hours after it is adherent after suck original culture
Base.Then blank group adds the culture mediums of RPMI 1640 containing 10% hyclone；Experimental group is separately added into containing 0.01~
The culture mediums of RPMI 1640 of 100 μ g/mL medicines to be measured；Continue after cultivating 48h, add concentration 5mg/mL MTT, continue to train
4h is supported, sucks supernatant, adds 100 μ L DMSO, 10min is placed in dark place, using ELIASA (Sunrise companies) under 570nm
Light absorption value is determined, and cell survival is calculated according to light absorption value, each processing sets 6 repeating holes.Cell survival rate (%)=
ΔODDrug-treated group/ΔODBlank group×100.Cell 503nhibiting concentration (IC is calculated further according to dose-effect curve50), it the results are shown in Table 1.It is real
The medicine difference for testing group is as follows：Experimental group 1 uses exclusive use Nobiletin；Kaempferol is used alone in experimental group 2；Experimental group 3
Rhoifolin is used alone；Experimental group 4 is 3 by weight using Nobiletin, Kaempferol and Rhoifolin:2:2 compositions
Composition 1；Experimental group 5 is 4 by weight using Nobiletin, Kaempferol and Rhoifolin:2:The composition 2 of 2 compositions；It is real
It is 4 by weight that group 6, which is tested, using Nobiletin, Kaempferol and Rhoifolin:3:The composition 2 of 3 compositions；Experimental group 7 uses
The Exocarpium Citri Grandis extract that embodiment 1 is prepared；The Exocarpium Citri Grandis extract that experimental group 8 is prepared using embodiment 2；Experimental group
9 Exocarpium Citri Grandis extracts being prepared using embodiment 3.
IC of the medicine of table 1. to human liver cancer cell BEL-740450Test result
From the experimental group 1~6 in table 1 as can be seen that Nobiletin, Kaempferol and Rhoifolin, which are used alone, has one
Fixed anti-liver cancer efficacy, when Nobiletin, Kaempferol and Rhoifolin three are mixed in ratio of the present invention, it is anti-
Liver cancer efficacy is more notable, and Synergistic anti-cancer effect can be produced after this explanation Nobiletin, Kaempferol and Rhoifolin three mixing.
Therefore, Nobiletin, Kaempferol and Rhoifolin three combination can be used as medicines resistant to liver cancer.
From the experimental group 7~9 in table 1 as can be seen that Exocarpium Citri Grandis extract of the present invention has the anti-of highly significant
Liver cancer acts on, and its antihepatocarcinoma effect is better than the antihepatocarcinoma effect of any of which monomer, this be due to it is therein effectively into
Nobiletin, Kaempferol and Rhoifolin is divided to produce the effect of collaboration anti-liver cancer and anti-.As can be seen here, the present invention have developed first
With very excellent anti-liver cancer efficacy and the clear and definite Exocarpium Citri Grandis extract of active ingredient, the extract can be used as anti-liver cancer drug
Thing uses.
Claims (10)
1. a kind of Exocarpium Citri Grandis extract, it is characterised in that include following active ingredient：Nobiletin, Kaempferol and Rhus succedanea
Glycosides.
2. Exocarpium Citri Grandis extract according to claim 1, it is characterised in that comprising following percentage by weight it is effective into
Point：Nobiletin 30~40%；Kaempferol 20~30%；Rhoifolin 20~30%.
3. the preparation method of the Exocarpium Citri Grandis extract described in claim 1 or 2, it is characterised in that comprise the following steps：
(1) Exocarpium Citri Grandis is taken, with ethanol heating and refluxing extraction, concentrated extracting solution obtains Exocarpium Citri Grandis ethanol extract；
(2) Exocarpium Citri Grandis ethanol extract is dissolved in water into suspension, first with petroleum ether extraction, discards petroleum ether extract, then
It is extracted with ethyl acetate, concentration acetic acid ethyl ester extract obtains Exocarpium Citri Grandis extract；
(3) Exocarpium Citri Grandis extract is produced into Exocarpium Citri Grandis extract through silica gel column chromatography is active constituent-enriched.
4. preparation method according to claim 3, it is characterised in that the ethanol described in step (1) is that volume fraction is
80~100% ethanol water；The dosage of described ethanol and the amount ratio of Exocarpium Citri Grandis are 8~15mL:1g；Described heating
The time of refluxing extraction is 1~3h.
5. preparation method according to claim 3, it is characterised in that first with the stone isometric with suspension in step (2)
Oily ether extracts 1~3 time, discards petroleum ether extract, then is extracted 1~3 time with the isometric ethyl acetate of suspension, concentrates second
Acetoacetic ester extract obtains Exocarpium Citri Grandis extract.
6. preparation method according to claim 3, it is characterised in that silica gel column chromatography is active constituent-enriched in step (3)
Specific method be：It is first 90 with volume ratio by silicagel column on Exocarpium Citri Grandis extract:10~87:13 solvent orange 2 A and solvent B groups
Into mixed solvent system carry out elution removal of impurities；Then it is 82 to use volume ratio again:18~80:20 solvent orange 2 A and solvent B composition
Mixed solvent system is eluted and collects elution position, produces Exocarpium Citri Grandis active component；Described solvent orange 2 A is chloroform, solvent B
For methanol；
Volume ratio is 90:10~87:The dosage of the mixed solvent system of 13 solvent orange 2 A and solvent B composition is silicagel column column volume
3~5 times；Volume ratio is 82:18~80:The dosage of the mixed solvent system of 20 solvent orange 2 A and solvent B composition is silicagel column post
5~8 times of volume.
7. preparation method according to claim 6, it is characterised in that silica gel column chromatography is active constituent-enriched in step (3)
Specific method be：It is first 87 with volume ratio by silicagel column on Exocarpium Citri Grandis extract:13 solvent orange 2 A：The mixing of solvent B compositions
Dicyandiamide solution carries out elution removal of impurities；Then it is 82 to use volume ratio again:18 solvent orange 2 A：The mixed solvent system of solvent B compositions is carried out
Elute and collect elution position, produce Exocarpium Citri Grandis active component；Described solvent orange 2 A is chloroform, and solvent B is methanol；
Volume ratio is 87:The dosage of the mixed solvent system of 13 solvent orange 2 A and solvent B composition is 3 times of silicagel column column volume；Body
Product is than being 82:The dosage of the mixed solvent system of 18 solvent orange 2 A and solvent B composition is 8 times of silicagel column column volume.
8. preparation method according to claim 6, it is characterised in that the weight of used silica gel extracts for Exocarpium Citri Grandis in silicagel column
Take 20~40 times of thing dry weight.
9. application of the Exocarpium Citri Grandis extract described in claim 1 or 2 in cancer therapy drug is prepared；Described cancer therapy drug is anti-
Liver-cancer medicine.
A kind of 10. anticancer pharmaceutical composition from Exocarpium Citri Grandis, it is characterised in that comprising following percentage by weight it is effective into
Point：Nobiletin, Kaempferol, the weight ratio of Rhoifolin are 3~4：2~3：2~3；Described cancer therapy drug is anti-liver cancer drug
Thing.
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| CN111643564A (en) * | 2020-07-17 | 2020-09-11 | 广东省农业科学院茶叶研究所 | Application of pummelo peel tea in preparation of anti-liver cancer drugs |
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| CN111643564A (en) * | 2020-07-17 | 2020-09-11 | 广东省农业科学院茶叶研究所 | Application of pummelo peel tea in preparation of anti-liver cancer drugs |
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