CN116621891A - Dictamni-containing phenolic glycoside C and preparation method and application thereof - Google Patents
Dictamni-containing phenolic glycoside C and preparation method and application thereof Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 229930182487 phenolic glycoside Natural products 0.000 title claims abstract description 20
- 150000007950 phenolic glycosides Chemical class 0.000 title claims abstract description 20
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims abstract description 63
- 235000018274 Cunila origanoides Nutrition 0.000 claims abstract description 28
- 235000014866 Dictamnus albus Nutrition 0.000 claims abstract description 28
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 10
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 201000007270 liver cancer Diseases 0.000 claims abstract description 9
- 208000014018 liver neoplasm Diseases 0.000 claims abstract description 9
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 claims abstract description 6
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 6
- 201000005249 lung adenocarcinoma Diseases 0.000 claims abstract description 6
- 239000002024 ethyl acetate extract Substances 0.000 claims abstract description 5
- 239000003208 petroleum Substances 0.000 claims abstract description 5
- 240000000774 Cunila origanoides Species 0.000 claims abstract 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 78
- 238000000605 extraction Methods 0.000 claims description 18
- WIONIXOBNMDJFJ-UHFFFAOYSA-N dictamnine Natural products C1=CC=C2C(OC)=C(C=CO3)C3=NC2=C1 WIONIXOBNMDJFJ-UHFFFAOYSA-N 0.000 claims description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 239000012046 mixed solvent Substances 0.000 claims description 12
- QZMAEZWZCGBZFK-AOJWCAIYSA-N (2s,3s,4s,5r,6r)-6-[[(3s,4ar,6ar,6bs,8as,12as,14ar,14br)-4,4,6a,6b,11,11,14b-heptamethyl-8a-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxycarbonyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3,5-dihydroxy-4-[(2s,3r Chemical compound O([C@H]1[C@H](O)[C@H](O[C@H]([C@@H]1O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@]1(CCC(C[C@H]14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C(O)=O)[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O QZMAEZWZCGBZFK-AOJWCAIYSA-N 0.000 claims description 10
- QZMAEZWZCGBZFK-UHFFFAOYSA-N 28-(beta-D-Glucopyranosyloxy)-28-oxoolean-12-en-3beta-yl 3-O-(beta-D-glucopyranosyl)-beta-D-glucopyranosiduronic acid Natural products C12CC(C)(C)CCC2(C(=O)OC2C(C(O)C(O)C(CO)O2)O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC(C1O)OC(C(O)=O)C(O)C1OC1OC(CO)C(O)C(O)C1O QZMAEZWZCGBZFK-UHFFFAOYSA-N 0.000 claims description 10
- KPJWZUAARPJYSB-UHFFFAOYSA-N glycoside C Natural products CC1(C)OC(=O)C23CCC1C2(O)CCC(C1(CCC24)C)(C)C3CCC1C2(C)CCCC4(C)COC(C(C(O)C1O)OC2C(C(O)C(CO)O2)O)OC1COC1OC(CO)C(O)C(O)C1O KPJWZUAARPJYSB-UHFFFAOYSA-N 0.000 claims description 10
- 239000002246 antineoplastic agent Substances 0.000 claims description 8
- 229940041181 antineoplastic drug Drugs 0.000 claims description 8
- 239000000469 ethanolic extract Substances 0.000 claims description 8
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 239000000725 suspension Substances 0.000 claims description 7
- 238000010521 absorption reaction Methods 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 239000003405 delayed action preparation Substances 0.000 claims description 5
- 239000004480 active ingredient Substances 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 239000007902 hard capsule Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 239000007901 soft capsule Substances 0.000 claims description 3
- 239000002552 dosage form Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 230000002459 sustained effect Effects 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 abstract description 8
- 230000005918 in vitro anti-tumor Effects 0.000 abstract description 4
- 238000002953 preparative HPLC Methods 0.000 abstract description 2
- 239000000284 extract Substances 0.000 abstract 1
- -1 methyl2- (2-hydroxy-6-methoxy-4- ((3, 4,5-trihydroxy-6- (hydroxy-methyl) tetra-hydro-2H-pyran-2-yl) oxy) phenyl) -4-methoxybenzoate Chemical compound 0.000 abstract 1
- 238000010298 pulverizing process Methods 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 244000182625 Dictamnus albus Species 0.000 description 20
- 239000003814 drug Substances 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 5
- 201000005202 lung cancer Diseases 0.000 description 5
- 208000020816 lung neoplasm Diseases 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
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- 239000000243 solution Substances 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 241000123855 Dictamnus dasycarpus Species 0.000 description 1
- VHLJDTBGULNCGF-UHFFFAOYSA-N Limonin Natural products CC1(C)OC2CC(=O)OCC23C4CCC5(C)C(CC(=O)C6OC56C4(C)C(=O)CC13)c7cocc7 VHLJDTBGULNCGF-UHFFFAOYSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 241001093501 Rutaceae Species 0.000 description 1
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- 230000003110 anti-inflammatory effect Effects 0.000 description 1
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- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000003560 cancer drug Substances 0.000 description 1
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- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- KBDSLGBFQAGHBE-MSGMIQHVSA-N limonin Chemical compound C=1([C@H]2[C@]3(C)CC[C@H]4[C@@]([C@@]53O[C@@H]5C(=O)O2)(C)C(=O)C[C@@H]2[C@]34COC(=O)C[C@@H]3OC2(C)C)C=COC=1 KBDSLGBFQAGHBE-MSGMIQHVSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/203—Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
Dictamni-side C (dascarpusolside C), a preparation method and application thereof, and relates to a novel compound, a preparation method and application thereof. The invention aims to provide a dittany phenolic glycoside C (dascarpusolside C), a preparation method and application thereof. Dictamni-glycoside C is known as methyl2- (2-hydroxy-6-methoxy-4- ((3, 4,5-trihydroxy-6- (hydroxy-methyl) tetra-hydro-2H-pyran-2-yl) oxy) phenyl) -4-methoxybenzoate, and has a molecular formula of C 22 H 26 O 12 The molecular weight is 482.1424. The preparation method comprises the following steps: 1. pulverizing cortex Dictamni Radicis, and extracting with ethanol; 2. dispersing the ethanol-recovered extract with water, and sequentially extracting with petroleum ether, dichloromethane and ethyl acetate; 3. and (3) repeatedly performing silica gel column chromatography on the ethyl acetate extract, and performing preparative high performance liquid chromatography separation. The invention relates to a dittany phenolic glycosideC (dascarpusolside C) has strong in vitro anti-tumor activity, and has good in vitro anti-tumor activity with IC50 of 13.72 μm/L and 15.20 μm/L for inhibiting lung adenocarcinoma cells (A549) and liver cancer tissue cells (HepG 2), respectively.
Description
Technical Field
The invention relates to a novel compound, a preparation method and application.
Background
Lung cancer is one of the common malignant tumors in China, and the incidence rate and death rate of lung cancer are the first. The lung cancer is hidden, most patients have metastasis during diagnosis, molecular targeted therapy and immunotherapy have great breakthroughs, traditional Chinese medicine plays an important role in the field of treating lung cancer, and the excavation of single medicines for treating lung cancer from traditional Chinese medicine is also the key content in the field of pharmacy.
Hepatocellular carcinoma is one of the common cancers and is also the third leading cause of cancer-related death. Liver cancer is receiving great attention due to its high incidence, high malignancy, strong invasion and metastasis, and poor prognosis. In recent years, the incidence rate of liver cancer continuously rises worldwide, and in terms of treatment, the local and systemic treatment is mainly carried out, and the traditional Chinese medicine has better curative effect in the aspect of treating liver cancer. Finding anti-liver cancer drugs from natural drugs is also a hotspot field of natural drug chemistry research.
The cortex Dictamni is dried root bark of Dictamni (Dictamnus dasycarpus Turcz.) belonging to Rutaceae. The chemical components mainly comprise limonin components, alkaloids and the like, and the cortex dictamni has various biological activities such as anti-inflammatory, antibacterial, antitumor and other activities, but specific anticancer medicinal components are not clear. There are many unknown compounds for the cortex dictamni yet to be developed and there is a need to discover and identify more active compounds.
Disclosure of Invention
The invention aims to: the first object of the present invention is to provide a novel compound, dictamnin C (dascarpusolside C), having in vitro antitumor activity. The second object of the present invention is to provide a method for preparing dittany phenolic glycoside C.
The third object of the invention is to provide the application of the dictamnin C in resisting tumors.
The molecular formula of the dittany phenolic glycoside C is C 22 H 26 O 12 The molecular weight is 482.1424, and the molecular structural formula is:
the preparation method of the dittany phenolic glycoside C (dascarpusolside C) comprises the following steps:
(1) Crushing cortex dictamni, and placing the crushed cortex dictamni into an extraction tank of a Soxhlet dynamic extraction concentration unit to extract the cortex dictamni with ethanol with the volume concentration of 95% to obtain ethanol extract;
(2) Concentrating the ethanol extract obtained in the step (1), dispersing with water to obtain a suspension, and then sequentially extracting with petroleum ether, dichloromethane and ethyl acetate;
(3) Subjecting the ethyl acetate extract obtained in the step (2) to silica gel column chromatography, sequentially eluting with mixed solvents of dichloromethane and methanol in volume ratios of 100:0, 100:1, 100:2, 100:5 and 100:10, subjecting the eluate with the dichloromethane and methanol in volume ratio of 100:10 to reverse phase silica gel (ODS, 50 μm, G1P 4S6 CANADA) column chromatography, sequentially eluting with mixed solvents of methanol and water in volume ratios of 10:90 and 20:80, subjecting the eluate with the methanol and water in volume ratio of 20:80 to silica gel column chromatography, sequentially eluting with mixed solvents of dichloromethane and methanol in volume ratios of 100:10 and 100:20, collecting the eluate with the dichloromethane and methanol in volume ratio of 100:20, subjecting the eluate to preparative high performance liquid chromatography, and collecting absorption peaks occurring in the eluting time of 8.54 min to obtain the dictamnin C;
the high performance liquid chromatography prepared in the step (3) takes acetonitrile and water with volume ratio of 10:90 as mobile phases, and the flow rate is 3ml/min.
And (3) setting the pressure of the Soxhlet dynamic extraction concentration unit in the step (1) as normal pressure extraction, and setting the extraction temperature at 85-90 ℃.
The concentration condition of the step (2) is-0.05 to-0.08 MPa.
The suspension density in step (2) was 1.25g/ml.
The application of the dittany phenol glycoside C is that the dittany phenol glycoside C is used for preparing antitumor drugs.
The preparation form of the antitumor drug comprises injection, freeze-dried powder injection or oral preparation.
The oral preparation comprises tablets, granules, soft capsules, hard capsules, oral liquid or sustained and controlled release preparations.
The dittany phenol glycoside C is used as an active ingredient of an anti-tumor medicament for inhibiting liver cancer cells and lung adenocarcinoma cells.
The dittany phenolic glycoside C is colorless crystal and is dissolved in methanol.294。IR：3394.0(cm -1 ) Is a hydroxyl absorption peak; 1707.2 (cm) -1 ) Is the carbonyl absorption peak. HR-ESI-MS m/z 505.1310[ M+Na ]] + (calculated 505.1316, ppm=1.18), molecular formula C was determined 22 H 26 O 12 The calculated unsaturation was 10.
The beneficial effects are that: compared with the prior art, the invention has the following advantages:
1. the invention obtains the new compound dittany phenol glycoside C for the first time, and the purity of the dittany phenol glycoside C obtained by the preparation method is high and reaches more than 98.4 percent.
2. The dictamnin C has stronger in-vitro anti-tumor activity, and has the IC50 of 13.72 mu M/L and 15.20 mu M/L for inhibiting lung adenocarcinoma cells (A549) and liver cancer tissue cells (HepG 2) respectively.
Detailed Description
The technical scheme of the invention is not limited to the specific embodiments listed below, and also includes any combination of the specific embodiments.
The first embodiment is as follows: the molecular formula of the dittany phenolic glycoside C in the embodiment is C 22 H 26 O 12 The molecular weight is 482.1424, and the molecular structural formula is:
the second embodiment is as follows: the preparation method of the dittany phenolic glycoside C in the embodiment is carried out according to the following steps:
(1) Crushing 100kg of cortex dictamni, and placing the crushed cortex dictamni into an extraction tank of a Soxhlet dynamic extraction concentration unit to extract the crushed cortex dictamni with 95% ethanol by volume concentration to obtain ethanol extract;
(2) Concentrating the ethanol extract obtained in the step (1), dispersing with water to obtain a suspension, and then sequentially extracting with petroleum ether, dichloromethane and ethyl acetate;
(3) Subjecting the ethyl acetate extract obtained in the step (2) to silica gel column chromatography, eluting with mixed solvents with the volume ratio of dichloromethane to methanol of 100:0, 100:1, 100:2, 100:5 and 100:10 in sequence, subjecting the eluate with the volume ratio of dichloromethane to methanol of 100:10 to reverse phase silica gel (ODS, 50 μm, G1P 4S6 CANADA) column chromatography, eluting with mixed solvents with the volume ratio of methanol to water of 10:90 and 20:80 in sequence, subjecting the eluate with the volume ratio of methanol to 20:80 to silica gel column chromatography, eluting with mixed solvents with the volume ratio of dichloromethane to methanol of 100:10 and 100:20 in sequence, collecting the eluate with the volume ratio of dichloromethane to methanol of 100:20, performing preparation high performance liquid chromatography, and collecting absorption peaks occurring in the elution time of 8.54 min to obtain the dictamnin C;
and (3) preparing the high performance liquid chromatography by taking acetonitrile and water with volume ratio of 10:90 as mobile phases, wherein the flow rate is 3ml/min.
And a third specific embodiment: the second difference between the present embodiment and the specific embodiment is that the pressure of the soxhlet dynamic extraction and concentration unit in the step (1) is set to normal pressure extraction, and the extraction temperature is set to 85-90 ℃. The other is the same as in the second embodiment.
The specific embodiment IV is as follows: the difference between the present embodiment and the second to third embodiments is that the concentration condition in the step (2) is-0.05 to-0.08 MPa. The other is the same as in one of the second to third embodiments.
Fifth embodiment: this embodiment differs from the second to fourth embodiments in that the suspension density in step (2) is 1.25g/ml. The others are the same as in the second to fourth embodiments.
Specific embodiment six: the application of the dittany phenol glycoside C in the preparation of antitumor drugs is provided in the specific embodiments one to five.
Seventh embodiment: the sixth difference between the present embodiment and the specific embodiment is that the dosage form of the antitumor drug includes injection, lyophilized powder for injection and oral preparation. The other is the same as in the sixth embodiment.
Eighth embodiment: the sixth difference between the present embodiment and the specific embodiment is that the oral preparation includes a tablet, a granule, a soft capsule, a hard capsule, an oral liquid or a sustained-release preparation. The other is the same as in the sixth embodiment.
Detailed description nine: the sixth embodiment differs from the sixth embodiment in that dictamnin C is used as an active ingredient of an antitumor drug for inhibiting hepatoma cells and lung adenocarcinoma cells. The other is the same as in the sixth embodiment.
The following experiments are adopted to verify the effect of the invention:
experiment one:
the preparation method of the dittany phenolic glycoside C comprises the following steps:
(1) Crushing 100kg of cortex dictamni, and placing the crushed cortex dictamni into an extraction tank of a Soxhlet dynamic extraction concentration unit to extract the crushed cortex dictamni with 95% ethanol by volume concentration to obtain ethanol extract;
(2) Concentrating the ethanol extract obtained in the step (1), dispersing with water to obtain a suspension with the density of 1.25g/ml, and then sequentially extracting with petroleum ether, dichloromethane and ethyl acetate;
(3) Subjecting the ethyl acetate extract obtained in the step (2) to silica gel column chromatography, sequentially eluting with mixed solvents of dichloromethane and methanol in a volume ratio of 100:0, 100:1,100:5 and 100:10, subjecting the eluate with the dichloromethane and methanol in a volume ratio of 100:10 to reverse phase silica gel (ODS, 50 μm, G1P 4S6 CANADA) column chromatography, sequentially eluting with mixed solvents of methanol and water in a volume ratio of 10:90 and 20:80, subjecting the eluate with methanol and water in a volume ratio of 20:80 to silica gel column chromatography, eluting with mixed solvents of dichloromethane and methanol in a volume ratio of 100:10 and 100:20, collecting the eluate with the dichloromethane and methanol in a volume ratio of 100:20, preparing high performance liquid chromatography, and collecting absorption peaks occurring in the eluting time of 8.54 min to obtain the monomer compound, which is identified as a new compound, named as dictamnide C. And (3) preparing the high performance liquid chromatography by taking acetonitrile and water with volume ratio of 10:90 as mobile phases, wherein the flow rate is 3ml/min.
Wherein the extraction conditions of the cortex dictamni and 95% ethanol in the step (1) are as follows: the normal pressure and the temperature are 90 ℃.
Wherein the concentration condition in the step (2) is-0.05 MPa.
The experiment adopts a Waters2545 preparation type high performance liquid chromatography, the detector is a Waters2489 type ultraviolet detector, and the detection wavelength is 210nm. The packing of the chromatographic column is C-18 reverse phase silica gel.
100kg of dittany bark is crushed and extracted for the experiment, and finally, 6mg of dittany phenolic glycoside C is obtained, the purity reaches 98.4%, and nuclear magnetic resonance data of the dittany phenolic glycoside C are shown in table 1.
TABLE 1 Dictamni phenolic glycoside C 1 H-NMR (600 MHz, MEOD) 13 C-NMR(150MHz,MEOD)
Experiment II:
the inhibition effect of the dittany phenolic glycoside C prepared in the experiment one on lung adenocarcinoma cells (A549) and liver cancer tissue cells (HepG 2) is detected:
1. a549 cells and HepG2 cells in logarithmic growth phase were cultured at 1X 10 5 Wells were seeded in 96-well plates in 5% CO 2 Culturing in an incubator at 37 ℃ for 24 hours.
2. Preparing the dittany phenol glycoside C prepared in the first experiment into a DMSO solution with the concentration of 600 mu M/L, and then carrying out gradient dilution by using a culture medium, wherein the concentration of a sample group is set to be 60 mu M/L, 30 mu M/L, 15 mu M/L, 7.5 mu M/L and 3.75 mu M/L; the blank group was set as a blank medium without dittany phenolic glycoside C.
3. And (3) adding the drug in the second step into the A549 cells and the HepG2 cells which are treated in the first step for 4 hours, discarding the supernatant, adding the MTT solution, and detecting the cell inhibition rate. Cell inhibition (%) = (MTT blank OD 570nm MTT sample group OD 570nm ) MTT blank OD 570nm 。
4. The procedure was repeated three times in succession, and the IC50 value was calculated by Logit method. IC50 of the dittany phenol glycoside C prepared in the experiment I for inhibiting A549 and HepG2 cells is measured to be 13.72 mu M/L and 15.20 mu M/L respectively.
Claims (9)
1. The dittany phenolic glycoside C is characterized in that the molecular formula of the dittany phenolic glycoside C is C 22 H 26 O 12 The molecular weight is 482.1424, and the molecular structural formula is:
2. the method for preparing the dictamnin C as claimed in claim 1, which is characterized by comprising the following steps:
(1) Crushing cortex dictamni, and placing the crushed cortex dictamni into an extraction tank of a Soxhlet dynamic extraction concentration unit to extract the cortex dictamni with ethanol with the volume concentration of 95% to obtain ethanol extract;
(2) Concentrating the ethanol extract obtained in the step (1), dispersing with water to obtain a suspension, and then sequentially extracting with petroleum ether, dichloromethane and ethyl acetate;
(3) Subjecting the ethyl acetate extract obtained in the step (2) to silica gel column chromatography, sequentially eluting with mixed solvents with the volume ratio of dichloromethane to methanol of 100:0, 100:1, 100:2, 100:5 and 100:10, subjecting the eluate with the volume ratio of dichloromethane to methanol of 100:10 to reverse phase silica gel column chromatography, sequentially eluting with mixed solvents with the volume ratio of methanol to water of 10:90 and 20:80, subjecting the eluate with the volume ratio of methanol to silica gel column chromatography, subjecting the eluate with the volume ratio of methanol to 20:80 to silica gel column chromatography, sequentially eluting with mixed solvents with the volume ratio of dichloromethane to methanol of 100:10 and 100:20, collecting the eluate with the volume ratio of dichloromethane to methanol of 100:20, and collecting the absorption peak occurring in the eluting time of 8.54 min to obtain the dictaphenol glycoside C;
the high performance liquid chromatography prepared in the step (3) takes acetonitrile and water with volume ratio of 10:90 as mobile phases, and the flow rate is 3ml/min.
3. The method for preparing dittany phenolic glycoside C according to claim 2, characterized in that the Soxhlet dynamic extraction and concentration unit pressure in step (1) is set to normal pressure extraction, and the extraction temperature is set to 85-90 ℃.
4. The method for preparing dittany phenolic glycoside C according to claim 2, characterized in that the concentration condition in step (2) is-0.05 to-0.08 MPa.
5. The method for preparing dittany phenolic glycoside C according to claim 2, characterized in that the suspension density in step (2) is 1.25g/ml.
6. The use of the dictamnin C as claimed in claim 1, characterized in that the dictamnin C is used for preparing antitumor drugs.
7. The application of the dittany phenol glycoside C according to claim 6, which is characterized in that the dosage forms of the antitumor drugs comprise injection, freeze-dried powder injection and oral preparation.
8. The use of the dictamnine C as claimed in claim 7, wherein the oral preparation comprises tablets, granules, soft capsules, hard capsules, oral liquid and sustained and controlled release preparations.
9. The use of the dictamnin C as claimed in claim 6, characterized in that the dictamnin C is used as an active ingredient of an antitumor drug for inhibiting liver cancer cells and lung adenocarcinoma cells.
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