CN105017357B - Polyphenol chromocor class compound and its preparation method and application - Google Patents
Polyphenol chromocor class compound and its preparation method and application Download PDFInfo
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Abstract
The invention discloses polyphenol chromocor class compound of one kind such as formula (I) and its preparation method and application,Formula (I), wherein R1、R3Can simultaneously be H or OH or in which 1；R2、R4And R5It is glycosyl wherein at least to have 1, remaining is H or OH；Glycosyl type has glucose (Glc), galactolipin (Gala), mannose (Man), rhamnose (Rham)；It can also connect to form bioside or three glucosides between sugar, it is the flavone compound containing polyphenol structure extracted from Chinese chestnut involucre, chestnut flower, chestnut shell, Chinese chestnut Seed, it can be combined with pharmaceutically acceptable carrier with preferable anti-human lung cancer, human liver cancer, human pancreas cancer, human gastric cancer and human colon carcinoma activity and prepare clinically acceptable drug, be used for oncotherapy.
Description
Technical field
The present invention relates to pharmaceutical technology fields, and in particular to a kind of polyphenol chromocor class compound and preparation method thereof and answers
With.
Background technology
Chinese chestnut (Castanea mollissima Blume) also known as chestnut, chestnut, wind wax are Fagaceae (Fagceae) Castaneas
(Castanea) plant, Chinese chestnut kernel are known as the good reputation of " king of dry fruit ", are China's specialties, and in foreign countries, it is also known as " ginseng
Fruit ".Chinese medicine thinks that Chinese chestnut is warm-natured, hides sweet flat, enters spleen, stomach, kidney channel.About the medical value of Chinese chestnut, Chinese medicine thinks, Li Guoyou
Nourishing stomach and spleen, Kidney-invigorating and tendon-reinforcing, the function of promoting blood circulation and hemostasis, and it is beneficial to the prevention of hypertension, coronary heart disease.Clinically, Chinese chestnut is also
It can be used for treating the diseases such as gastric disorder causing nausea, diarrhea, waist-leg is weak, spit blood, have blood in stool, metal-inflicted wound.Meanwhile Chinese chestnut is former plus other Chinese medicines or food
Herbal cuisine, treatment tracheitis, kidney deficiency, the diseases such as indigestion, diarrhea, apoplexy can be made in material.The pharmacological action of Chinese chestnut is related to
The various aspects such as anticoagulation, increasing leukocyte, treatment bacillary dysentery, treatment chronic bronchitis.
Phenolic acid compound is rich in Castanea, main chemical composition is all more in Chinese chestnut involucre, titbit, shell
Phenol.Our early periods isolated several polyphenol chromocor class compounds, structure from Chinese chestnut Seed, chestnut flower, Chinese chestnut involucre, chestnut shell
Flavonoid glycoside compound structure novel containing cinnamyl, coumaric acyl, asafoetide acyl group, gallic acid acyl group, antitumor
In activity rating, there is inhibitory activity, the constituents and compound, which can be used as, one or more can be used as treatment or complementary control
Treat the drug of cancer.
Invention content
To solve the above problems, the present invention provides a kind of polyphenol chromocor class compound and its preparation method and application, system
Standby process is simple and easy to do, and prepared component, monomeric compound have the antitumor activity of wide spectrum.
To achieve the above object, the technical solution that the present invention takes is：
Polyphenol chromocor class compound shown in a kind of formula (I),
Wherein R1、R3Can simultaneously be H or OH or in which 1；R2、R4And R5It is glycosyl wherein at least to have 1, remaining is H
Or OH；Glycosyl type has glucose (Glc), galactolipin (Gala), mannose (Man), rhamnose (Rham)；Can also it connect between sugar
It connects to form bioside or three glucosides, connection type has 1-2 connections, 1-3 connections, 1-4 connections to connect with 1-6.The C-2 of the sugar
Position, C-3, C-4 and C-6 can also connect formula (II) or formula (III),
R in formula (II) and formula (III)6、R7And R8Can be H, OH, OCH simultaneously with identical meaning3Or H, OH, OCH3
One or two of.
The present invention also provides three kinds of preparation methods of above compound：
Method one, includes the following steps：
S11, it takes medicinal material coarse powder using ultrasound or solvent heating extraction, water decoction or 30%-100% is measured using 3-15 times
Alcohol reflux extracts or the extraction of 30%-100% methanol eddies, water or alcoholic solvent is recovered under reduced pressure, residue is waved to no alcohol taste or alcoholic degree
After 10%, with centrifuging and taking supernatant after water dissolution.
Gained supernatant polyamide or macroporous resin treatment are enriched with 30%- by S12 with water-alcohol solution gradient elution
80% alcohol elution fraction, obtains crude flavonoid powder；
S13, gained crude flavonoid powder is detached through silica gel column chromatography, gradient elution, knot is carried out using different solvents system
Mesolow preparative separation chromatography, high performance liquid chromatography progress preparative separation are closed, is divided with acetonitrile-water system, methanol-water system
From obtaining polyphenol flavone compound.
Method two, includes the following steps：
S21, take medicinal material coarse powder using ultrasound or solvent heating and refluxing extraction method, the 30%-100% second measured using 3-10 times
Alcohol or methanol eddy extraction, are recovered under reduced pressure alcoholic solvent, after residue is waved to no alcohol taste or alcoholic degree less than 10%, with water dissolution, obtain mixed
Suspension；
S22, gained suspension is extracted with ethyl acetate, n-butanol successively, extraction times 1-5 times, extracts volume
(1: 1-1: 8) each extraction position is obtained, solvent is separately recovered, obtains medicinal extract, wherein ethyl acetate layer medicinal extract is thick total-flavonoid aglycone
Component；N-butanol layer is total polyphenols flavonoid glycoside；
S23, gained medicinal extract is detached through silica gel column chromatography, gradient elution is carried out using different solvents system, in conjunction with
Mesolow preparative separation chromatography, high performance liquid chromatography carry out preparative separation, are divided with acetonitrile/water system, methanol/water system
From obtaining polyphenol flavone compound.
Method three, includes the following steps：
S31, it takes medicinal material coarse powder using ultrasound or solvent heating and refluxing extraction method, ethyl acetate or acetone is measured using 3-10 times
Refluxing extraction is recovered under reduced pressure solvent and obtains medicinal extract, is thick general flavone；
S32, the thick general flavone of gained is detached through silica gel column chromatography, using petroleum ether, ethyl acetate, dichloromethane
The different solvents systems of the compositions such as alkane, chloroform, acetone, methanol carries out gradient elution, in conjunction with mesolow preparative separation chromatography,
High performance liquid chromatography carries out preparative separation, is detached with acetonitrile-water system, methanol-water system, obtains polyphenol flavonoid
Object.
Wherein, a concentration of 10%-100% of the methanol, a concentration of 10%-100% of the ethyl alcohol.
Wherein, the polyamide is one kind in PA6, PA66, PA11, PA12, PA46, PA610, PA612, PA1010
Or several mixtures, the macroreticular resin include but not limited to polarity, non-polar macroporous resin D-101, DM-301, HPD-
100、AB-8。
Wherein, the different solvents system includes but not limited to petroleum ether, ethyl acetate, dichloromethane, chloroform, third
Ketone, methanol.
Wherein, the medicinal material coarse powder is Chinese chestnut involucre, chestnut flower, chestnut shell, Chinese chestnut branch using Chinese medicines or plants such as Castaneas
One kind in item, Chinese chestnut Seed.
Wherein, mobile phase is methanol/water or acetonitrile/water or ethanol/water in standby separation chromatography, high performance liquid chromatography, is mixed
Composition and division in a proportion example is 1: 10-90: 10 or methanol/acetonitrile/water, mixed proportion 1: 1: 10-40: 40: 20.
On compound and thick general flavone can be used to prepare the drug of antitumor cell growth, the tumour be human lung cancer,
One kind in human liver cancer, human pancreas cancer, human gastric cancer and human colon carcinoma, while a kind of pharmaceutical composition can also be prepared for treating
Cancer, the composition include compound, thick general flavone and the pharmaceutically acceptable carrier for the treatment of effective dose.
The invention has the advantages that：
Preparation process is simple and easy to do, and prepared component, monomeric compound have the antitumor activity of wide spectrum.
Specific implementation mode
In order to make objects and advantages of the present invention be more clearly understood, the present invention is carried out with reference to embodiments further
It is described in detail.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to limit this hair
It is bright.
Embodiment 1
Thick polyphenol chromocor 1 is detached from Chinese chestnut
Chestnut involucre, chestnut flower, chestnut shell or each 2kg of Chinese chestnut Seed coarse powder are taken respectively, are carried with 8 times of 70% alcohol refluxs of amount
It takes 3 times, each 2h, ethyl alcohol is recovered under reduced pressure, residue is waved to no alcohol taste, with centrifuging and taking supernatant after water dissolution.
The supernatant open column chromatography processing of 2Kg macroporous absorbent resins (AB-8), with water-alcohol solution gradient elution, enrichment
30%-80% alcohol elution fractions, respectively obtain crude flavonoid powder 180g, 167g, 110g, 109g.
Embodiment 2
Crude flavonoid powder 2 is detached from Chinese chestnut
Medicinal material coarse powder i.e. each 0.5kg of chestnut involucre, chestnut flower, chestnut shell, Chinese chestnut Seed is taken, with 6 times of 70% EtOH Sonicates of amount
1hr is extracted, alcoholic solvent is recovered under reduced pressure, residue, which is waved to no alcohol taste or alcoholic degree, is less than 5%, with centrifuging and taking supernatant after water dissolution.On
Clear liquid exploitation polyamide (PA66) column chromatography processing, is eluted, enrichment 30%-70% alcohol elutions portion with water-ethanol solution gradient
Point, respectively obtain crude flavonoid powder 27g, 23g, 18g, 16g.
Embodiment 3
Crude flavonoid powder 3 is detached from Chinese chestnut
Chestnut involucre, chestnut flower, chestnut shell or each 2kg of Chinese chestnut Seed coarse powder are taken respectively, are carried with 8 times of 70% alcohol refluxs of amount
It takes 3 times, each 2h, ethyl alcohol is recovered under reduced pressure, residue is waved to no alcohol taste, and each suspension 2L is obtained with water dissolution.Suspension according to this use etc.
The ethyl acetate and n-butanol of volume are extracted, each extraction 3 times, and each extraction position is separately recovered solvent and obtains medicinal extract, wherein second
Ethyl acetate layer medicinal extract is thick total-flavonoid aglycone component, respectively 45-67g.N-butanol layer medicinal extract is total polyphenols flavonoid glycoside, respectively
For 61-102g, ethyl acetate and n-butanol layer component are merged to get crude flavonoid powder.
Embodiment 4
Crude flavonoid powder 4 is detached from Chinese chestnut
Chestnut involucre, chestnut flower, chestnut shell or each 2Kg of Chinese chestnut Seed coarse powder are taken respectively, and acetone heating and refluxing extractions are measured with 8 times,
Each 2h, is recovered under reduced pressure solvent, can obtain crude flavonoid powder 210-320g.
Embodiment 5
Polyphenol chromocor class compound is detached from Chinese chestnut crude flavonoid powder
Thick general flavone 100g in above-described embodiment 1 is taken, is detached through the open column chromatography of silica gel (100-300 mesh), with chlorine
It is imitative：Methanol (100: 0-1: 100) carries out gradient elution, and gained fraction merges same composition, obtain 10 through thin-layer chromatographic analysis
Fraction.Fraction 1-10 is handled through medium and low pressure column chromatographic isolation respectively, according to this with methanol: water (1: 9；2:8；3:7；6:4；5: 5) into
It goes and elutes, assistant merges identical fraction with thin-layer chromatographic analysis, obtains each eluate after removing solvent, eluate is laggard through HPLC analyses
Prepared by preparative HPLC, chromatographic condition is methanol-water-formic acid (10: 90: 0.02-65: 35: 0.02), obtains 15 structures as follows
Compound.
Compound 1-15 structures are as follows
The hydrocarbon modal data of compound 1-15 Structural Identifications is as follows：
Compound 1：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molisch reactions are the positive.1H-NMR (600MHz,
DMSO-d6)δH：6.70 (1H, br s, H-6), 6.70 (1H, br s, H-8), 8.45 (2H, d, J=8.4Hz, H-2 ', 6 '),
7.16 (2H, d, J=8.4Hz, H-3 ', 5 '), 5.27 (1H, d, J=7.6Hz, H-1 "), 7.51 (2H, d, J=8.4Hz, H-
2 " ', 6 " '), 7.16 (2H, d, J=8.4Hz, H-3 " ', 5 " '), 7.84 (1H, d, J=16.2Hz, H-7 " '), 6.51 (1H, d, J
=16.2Hz, H-8 " ')13C-NMR (150MHz, DMSO-d6)δC：157.8 (C-2), 133.4 (C-3), 178.4 (C-4),
162.8 (C-5), 99.9 (C-6), 166.0 (C-7), 94.7 (C-8), 157.7 (C-9), 105.2 (C-10), 121.9 (C-1 '),
131.9 (C-2 ', 6 '), 116.1 (C-3 ', 5 '), 161.8 (C-4 '), 104.1 (C-1 "), 76.0 (C-2 "), 78.4 (C-3 "),
(71.3 C-4 "), 76.0 (C-5 "), 64.2 (C-6 "), 126.1 (C-1 " '), 130.8 (C-2 " ', 6 " '), 116.8 (C-3 " ',
5 " '), 161.4 (C-4 " '), 145.2 (C-7 " '), 114.9 (C-8 " '), 167.3 (C-9 " ')
Compound 2：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molisch reactions are the positive.1H NMR (600MHz,
DMSO-d6)δH：6.11 (1H, d, J=2.1Hz, H-6), 6.27 (1H, d, J=2.1Hz, H-8), 7.60 (1H, d, J=2.1Hz,
H-2 '), 6.79 (1H, m, J=8.4Hz, H-5 '), 7.57 (1H, dd, J1=8.4,2.1Hz, H-6 '), 5.25 (1H, d, J=
7.2Hz, H-1 "), 7.29 (2H, d, J=8.7Hz, H-2 " ', 6 " '), 6.79 (2H, H-3 " ', 5 " '), 6.06 (1H, d, J=
15.9Hz, H-7 " '), 7.39 (1H, d, J=15.9Hz；H-8″′).13C-NMR (150MHz, DMSO-d6)δC：157.8 (C-2),
133.4 (C-3), 178.1 (C-4), 162.9 (C-5), 100.0 (C-6), 165.9 (C-7), 94.8 (C-8), 159.1 (C-9),
105.6 (C-10), 123.1 (C-1 '), 117.4 (C-2 '), 145.9 (C-3 '), 149.7 (C-4 '), 115.9 (C-5 '), 123.4
(C-6 '), 103.9 (C-1 "), 75.7 (C-2 "), 78.1 (C-3 "), 71.8 (C-4 "), 75.9 (C-5 "), 64.4 (C-6 "),
127.1 (C-1 " '), 131.2 (C-2 " '), 116.8 (C-3 " '), 161.1 (C-4 " '), 131.3 (C-5 " '), 116.8 (C-
6 " '), 146.5 (C-7 " '), 114.8 (C-8 " '), 168.9 (C-9 " ')
Compound 3：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molisch reactions are the positive.1H-NMR (400MHz,
CD3OD)δH：6.09 (1H, br s H-6), 6.27 (1H, br s, H-8), 7.99 (2H, d, J=9.0Hz, H-2 ', 6 '), 6.87
(2H, d, J=8.5Hz, H-3 ', 5 '), 5.67 (1H, d, J=7.5Hz, H-1 "), 5.24 (1H, br s, H-1 " '), 7.27 (2H,
D, J=8.0Hz, H-2 " ', 6 " '), 6.81 (2H, d, J=8.0Hz, H-3 " ', 5 " '), 7.38 (1H, d, J=16.0Hz, H-
7 " '), 6.03 (1H, d, J=16.0Hz, H-8 " ')13C-NMR (100MHz, CD3OD)δC：161.6 (C-2), 134.4 (C-3),
178.7 (C-4), 164.3 (C-5), 100.4 (C-6), 166.5 (C-7), 95.1 (C-8), 158.7 (C-9), 106.6 (C-10),
123.4 (C-1 '), 132.4 (C-2 ', 6 '), 116.4 (C-3 ', 5 '), 159.9 (C-4 '), 100.5 (C-1 "), 80.3 (C-2 "),
79.2 (C-3 "), 72.4 (C-4 "), 76.0 (C-5 "), 64.3 (C-6 "), 100.3 (C-1 " '), 72.7 (C-2 " '), 72.6 (C-
3 " '), 74.4 (C-4 " '), 70.3 (C-5 " '), 17.9 (C-6 " '), 127.5 (C-1 " "), 131.5 (C-2 " ", 6 " "), 117.2
(C-3 " ", 5 " "), 161.6 (C-4 " "), 146.9 (C-7 " ") and, 114.9 (C-8 " "), 169.1 (C-9 " ") and
Compound 4：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz, DMSO-
d6)δH：6.16 (1H, d, J=2.0Hz, H-6), 6.36 (1H, d, J=2.0Hz, H-8), 7.96 (2H, d, J=8.8Hz, H-2 ',
6 '), 6.87 (2H, d, J=8.8Hz, H-3 ', 5 '), 5.72 (1H, d, J=8.0Hz, H-1 "), 7.40 (2H, d, J=8.5Hz,
H-2 " ', 6 " '), 6.79 (2H, d, J=8.5Hz, H-3 " ', 5 " '), 7.35 (1H, d, J=15.8Hz, H-7 " '), 6.15 (1H,
D, J=15.8Hz, H-8 " '), 7.55 (2H, d, J=8.6Hz, H-2 " ", 6 " "), 6.79 (2H, d, J=8.6Hz, H-3 " ",
5 " "), 7.61 (1H, d, J=15.8Hz, H-7 " "), 6.43 (1H, d, J=15.8Hz, H-8 " ") and13C-NMR (150MHz,
DMSO-d6)δC：156.5 (C-2), 133.2 (C-3), 177.2 (C-4), 161.2 (C-5), 98.8 (C-6), 164.3 (C-7),
93.8 (C-8), 156.4 (C-9), 103.9 (C-10), 120.0 (C-1 '), 130.9 (C-2 ', 6 '), 115.2 (C-3 ', 5 '),
160.2 (C-4 '), 100.3 (C-1 "), 73.9 (C-2 "), 73.9 (C-3 "), 70.2 (C-4 "), 74.4 (C-5 "), 62.8 (C-
6 "), 125.0 (C-1 " '), 130.3 (C-2 " ', 6 " '), 115.9 (C-3 " ', 5 " '), 159.9 (C-4 " '), 144.8 (C-7 " '),
113.7 (C-8 " '), 166.2 (C-9 " '), 125.2 (C-1 " "), 130.4 (C-2 " ", 6 " "), 115.9 (C-3 " ", 5 " "),
160.1 (C-4 " "), 145.2 (C-7 " "), 114.3 (C-8 " ") and, 165.9 (C-9 " ")
Compound 5：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (400MHz, DMSO-
d6)δH：6.18 (1H, d, J=1.5Hz, H-6), 6.37 (1H, d, J=1.5Hz, H-8), 7.19 (2H, s, H-2 ', 6 '), 5.46
(1H, d, J=7.5Hz, H-1 "), 4.2 (1H, d, J=12Hz, Ha- 6 "), 4.3 (1H, dd, J1=12.0,5.0Hz, Hb- 6 "),
6.88 (2H, s, H-2 " ', H-6 " ')13C-NMR (100MHz, DMSO-d6)δC：155.9 (C-2, C-9), 133.5 (C-3),
177.7 (C-4), 161.1 (C-5), 98.1 (C-6), 164.0 (C-7), 93.9 (C-8), 103.2 (C-10), 121.1 (C-1 '),
108.9 (C-2 ', C-6 ', C-2 " ', C-6 " '), 145.0 (C-3 ', C-5 ', C-3 " ', C-5 " '), 136.1 (C-4 '), 103.9
(C-1 "), 73.1 (C-2 "), 70.0 (C-4 "), 76.1 (C-5 "), 64.0 (C-6 "), 120.8 (C-1 " '), 74.2 (C-3 "),
138.1 (C-4 " '), 165.9 (C-7 " ')
Compound 6：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz, C5D5N)
δH：6.69 (1H, d, J=2.2Hz, H-6), 6.74 (1H, d, J=2.2Hz, H-8), 8.26 (1H, d, J=2.2Hz, H-2 '),
7.23 (1H, d, J=8.2Hz, H-5 '), 3.81 (3H, s, OCH3), 7.38 (1H, dd, J1=8.2,2.2Hz, H-6 '), 5.23
(1H, d, J=7.5Hz, H-1 "), 7.71 (2H, s, H-2 " ', H-6 " ')13C-NMR (150MHz, C5D5N)δC：157.8 (C-2),
135.1 (C-3), 178.6 (C-4), 162.8 (C-5), 100.0 (C-6), 166.0 (C-7), 94.8 (C-8), 157.7 (C-9),
105.3 (C-10), 121.3 (C-1), 114.6 (C-2 '), 151.4 (C-3 '), 148.1 (C-4 '), 116.3 (C-5 '), 123.1
(C-6 '), 56.4 (OCH3), 104.7 (C-1 "), 76.0 (C-2 "), 78.4 (C-3 "), 71.2 (C-4 "), 76.2 (C-5 "),
64.2 (C-6 "), 121.0 (C-1 " '), 110.3 (C-2 " ', C-6 " '), 147.2 (C-3 " ', C-5 " '), 140.6 (C-4 " '),
167.0(C-7″′).
Compound 7：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (400MHz, CD3OD)
δH：6.56 (1H, d, J=2.3Hz, H-6), 6.87 (1H, d, J=2.3Hz, H-8), 7.64 (1H, d, J=2.1Hz, H-2 '),
6.80 (1H, m, J=8.4Hz, H-5 '), 7.58 (1H, dd, J18.4,2.1Hz, H-6 '), 5.86 (1H, d, J=7.9Hz, H-
1 "), 7.63 (2H, s, H-2 " '/6 " ')13C-NMR (100MHz, CD3OD)δC：158.2 (C-2), 135.1 (C-3), 178.4
(C-4), 163.1 (C-5), 99.2 (C-6), 166.1 (C-7), 94.5 (C-8), 158.8 (C-9), 105.6 (C-10), 123.0
(C-1 '), 117.2 (C-2 '), 145.7 (C-3 '), 149.7 (C-4 '), 115.9 (C-5 '), 123.6 (C-6 '), 100.0 (C-
1 "), 76.6 (C-2 "), 76.1 (C-3 "), 71.1 (C-4 "), 77.6 (C-5 "), 61.3 (C-6 "), 121.2 (C-1 " '), 111.0
(C-2 " ', 6 " '), 146.3 (C-3 " ', 5 " '), 140.2 (C-4 " '), 169.1 (C-7 " ')
Compound 8：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (400MHz, CD3OD)
δH：6.56 (1H, d, J=2.3Hz, H-6), 6.87 (1H, d, J=2.3Hz, H-8), 8.55 (2H, d, J=8.2Hz, H-2 ',
6 '), 7.66 (2H, d, J=8.2Hz, H-3 ', 5 '), 5.86 (1H, d, J=7.9Hz, H-1 "), 7.63 (2H, s, H-2 " '/
6″′).13C-NMR (100MHz, CD3OD)δC：158.2 (C-2), 133.1 (C-3), 178.4 (C-4), 163.1 (C-5), 99.2
(C-6), 166.1 (C-7), 94.5 (C-8), 158.8 (C-9), 105.6 (C-10), 123.0 (C-1 '), 133.0 (C-2 ', 6 '),
117.2 (C-3 ', 5 '), 162.7 (C-4 '), 100.0 (C-1 "), 76.6 (C-2 "), 76.1 (C-3 "), 71.1 (C-4 "), 77.6
(C-5 "), 61.3 (C-6 "), 121.2 (C-1 " '), 111.0 (C-2 " ', 6 " '), 146.3 (C-3 " ', 5 " '), 140.2 (C-
4 " '), 169.1 (C-7 " ')
Compound 9：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (400MHz, CD3OD)
δH：6.13 (1H, d, J=2.1Hz, H-6), 6.30 (1H, d, J=2.1Hz, H-8), 7.20 (2H, s, H-2 ', 6 '), 5.76
(1H, d, J=7.9Hz, H-1 "), 7.12 (2H, s, H-2 " ', 6 " ')13C-NMR (100MHz, CD3OD)δC：158.2 (C-2),
133.1 (C-3), 178.9 (C-4), 1 δ 3.1 (C-5), 99.9 (C-6), 166.3 (C-7), 94.6 (C-8), 158.4 (C-9),
(105.7 C-10), 122.2 (C-1 '), 109.9 (C-2 ', 6 '), 146.4 (C-3 ', 5 '), 137.9 (C-4 '), 100.7 (C-
1 "), 76.2 (C-2 "), 76.6 (C-3 "), 71.5 (C-4 "), 78.7 (C-5 "), 62.5 (C-6 "), 121.4 (C-1 " '), 110.6
(C-2 " ', 6 " '), 146.5 (C-3 " ', 5 " '), 140.0 (C-4 " '), 168.1 (C-7 " ')
Compound 10：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz,
C5D5N)δH：6.65 (1H, d, J=2.2Hz, H-6), 6.67 (1H, d, J=2.2Hz, H-8), 8.83 (1H, d, J=2.2Hz, H-
2 '), 7.21 (1H, d, J=8.2Hz, H-5 '), 3.81 (3H, s, OCH3 ), 7.80 (1H, dd, J=8.2,2.2Hz, H-6 '),
5.80 (1H, d, J=8.0Hz, H-1 "), 7.41 (2H, d, J=8.8Hz, H-2 " ', H-6 " '), 7.12 (2H, d, J=8.8Hz,
H-3 " ', H-5 " '), 7.70 (1H, d, J=16.0Hz, H-7 " '), 6.35 (1H, d, J=16.0Hz, H-8 " ')13C-NMR
(150MHz, C5D5N)δC：157.6 (C-2), 135.1 (C-3), 178.5 (C-4), 162.7 (C-5), 99.9 (C-6), 166.0
(C-7), 94.7 (C-8), 157.6 (C-9), 105.2 (C-10), 121.2 (C-1 '), 114.6 (C-2 '), 151.3 (C-3 '),
148.0 (C-4 '), 116.3 (C-5 '), 123.1 (C-6 '), 56.4 (OCH3), 104.7 (C-1 "), 76.0 (C-2 "), 78.3 (C-
3 "), 71.2 (C-4 "), 76.2 (C-5 "), 64.2 (C-6 "), 125.2 (C-1 " '), 131.0 (C-2 " ', C-6 " '), 116.1 (C-
3 " ', C-5 " '), 160.3 (C-4 " '), 144.5 (C-7 " '), 112.9 (C-8 " '), 167.1 (C-9 " ')
Compound 11：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (400MHz,
CD3OD)δH：6.29 (1H, br s H-6), 6.57 (1H, br s, H-8), 7.11 (1H, d, J=2.1Hz, H-2 '), 6.23 (1H,
M, J=8.4Hz, H-5 '), 7.12 (1H, dd, J=8.4,2.1Hz, H-6 '), 5.46 (1H, d, J=7.5Hz, H-1 "), 5.12
(1H, br s, H-1 " '), 7.60 (2H, s, H-2 " ', 6 " ')13C-NMR (100MHz, CD3OD)δC：158.1 (C-2), 133.9
(C-3), 178.7 (C-4), 163.4 (C-5), 100.1 (C-6), 166.2 (C-7), 95.1 (C-8), 158.5 (C-9), 106.9
(C-10), 122.9 (C-1 '), 116.9 (C-2 '), 145.1 (C-3 '), 149.5 (C-4 '), 116.2 (C-5 '), 100.1 (C-
1 "), 80.1 (C-2 "), 79.0 (C-3 "), 71.8 (C-4 "), 76.9 (C-5 "), 64.2 (C-6 "), 103.3 (C-1 " '), 72.1
(C-2 " '), 72.2 (C-3 " '), 74.8 (C-4 " '), 70.9 (C-5 " '), 18.2 (C-6 " '), 119.5 (C-1 " "), 108.1 (C-
2 " ", 6 " "), 145.4 (C-3 " ", 5 " "), 138.2 (C-4 " ") and, 165.0 (C-7 " ")
Compound 12：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz,
DMSO-d6)δH：6.05 (1H, br s, H-6), 6.24 (1H, br s, H-8), 7.89 (2H, d, J=8.8Hz, H-2 ', 6 '),
6.78 (2H, d, J=8.8Hz, H-3 ', 5 '), 5.57 (1H, d, J=8.0Hz, H-1 "), 7.34 (2H, d, J=8.4Hz, H-
2 " ', 6 " '), 6.70 (2H, d, J=8.4Hz, H-3 " ', 5 " '), 7.55 (1H, d, J=16.0Hz, H-7 " '), 6.25 (1H, d, J
=16.0Hz, H-8 " ')13C-NMR (150MHz, DMSO-d6)δC：158.1 (C-2), 134.6 (C-3), 178.8 (C-4),
162.8 (C-5), 100.4 (C-6), 165.4 (C-7), 94.5 (C-8), 158.0 (C-9), 105.7 (C-10), 122.6 (C-
1 '), 132.0 (C-2 ', 6 '), 116.0 (C-3 ', 5 '), 161.2 (C-4 '), 99.6 (C-1 "), 75.7 (C-2 "), 76.2 (C-
3 "), 71.5 (C-4 "), 78.6 (C-5 "), 62.5 (C-6 "), 127.1 (C-1 " '), 131.0 (C-2 " ' 6 " '), 116.6 (C-3 " '
5 " '), 160.9 (C-4 " '), 146.6 (C-7 " '), 115.0 (C-8 " '), 168.1 (C-9 " ')
Compound 13：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H NMR (400MHz,
CD3OD)：6.17 (1H, d, J=2.1Hz, H-6), 6.33 (1H, d, J=2.1Hz, H-8), 7.56 (1H, d, J=2.4Hz, H-
2 '), 6.72 (1H, d, J=9.0Hz, H-5 '), 7.54 (1H, m, H-6 '), 5.19 (1H, d, J=7.5Hz, H-1 "), δ 6.94
(2H, s, H-2 " ', 6 " ')13C NMR (100MHz, CD3OD)：δ：158.3 (C-2), 133.2 (C-3), 179.3 (C-4),
162.8 (C-5), 99.9 (C-6), 165.8 (C-7), 94.9 (C-8), 159.3 (C-9), 105.6 (C-10), 123.0 (C-1 '),
117.2 (C-2 '), 145.7 (C-3 '), 149.7 (C-4 '), 115.9 (C-5 '), 123.6 (C-6 '), 104.2 (C-1 "), 75.7
(C-2 "), 78.0 (C-3 "), 71.5 (C-4 "), 75.9 (C-5 "), 64.3 (C-6 "), 121.3 (C-1 " '), 110.2 (C-2 " ',
6 " '), 146.3 (C-3 " ', 5 " '), 139.7 (C-4 " '), 165.8 (C-7 " ')
Compound 14：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz,
DMSO-d6)δH：6.70 (1H, br s, H-6), 6.70 (1H, br s, H-8), 8.45 (2H, d, J=8.4Hz, H-2 ', 6 '),
7.16 (2H, d, J=814Hz, H-3 ', 5 '), 5.26 (1H, d, J=7.6Hz, H-1 "), 7.08 (1H, d, J=1.5Hz, H-
2 " '), 6.85 (1H, d, J=8.0Hz, H-5 " '), 6.99 (1H, dd, J=8.0,1.5Hz, H-6 " '), 7.54 (1H, d, J=
16.0Hz, H-7 " '), 6.24 (1H, d, J=16.0Hz, H-8 " ')13C-NMR (150MHz, DMSO-d6)δC：157.1 (C-2),
133.6 (C-3), 179.2 (C-4), 163.1 (C-5), 99.9 (C-6), 165.9 (C-7), 94.2 (C-8), 157.1 (C-9),
104.8 (C-10), 122.3 (C-1 '), 132.1 (C-2 ', 6 '), 116.0 (C-3 ', 5 '), 161.9 (C-4 '), 104.1 (C-
1 "), 76.5 (C-2 "), 73.2 (C-3 "), 71.3 (C-4 "), 76.0 (C-5 "), 64.2 (C-6 "), 127.1 (C-1 " '), 131.8
(C-2 " ', 6 " '), 116.1 (C-3 " ', 5 " '), 161.9 (C-4 " '), 145.0 (C-7 " '), 115.2 (C-8 " '), 167.0 (C-
9″′).
Compound 15：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz,
DMSO-d6)δH：6.70 (1H, br s, H-6), 6.71 (1H, br s, H-8), 8.45 (2H, d, J=8.4Hz, H-2 ', 6 '),
7.16 (2H, d, J=8.4Hz, H-3 ', 5 '), 5.23 (1H, d, J=7.6Hz, H-1 "), 7.53 (1H, d, J=16.0Hz, H-
7 " '), 6.24 (1H, d, J=16.0Hz, H-8 " ') .6.85 (1H, d, J=8.5Hz, H-5 " '), 6.99 (1H, dd, J=8.5,
2.5Hz, H-6 " '), 7.07 (1H, d, J=2.5Hz, H-2 " '), 3.78 (3H, s ,-OCH3)。13C-NMR (150MHz, DMSO-
d6)δC：157.8 (C-2), 133.3 (C-3), 178.7 (C-4), 162.8 (C-5), 100.0 (C-6), 166.0 (C-7), 94.7
(C-8), 157.7 (C-9), 105.2 (C-10), 121.9 (C-1 '), 131.9 (C-2 ', 6 '), 116.1 (C-3 ', 5 '), 161.8
(C-4 '), 104.1 (C-1 "), 76.0 (C-2 "), 78.4 (C-3 "), 71.3 (C-4 "), 76.0 (C-5 "), 64.2 (C-6 "),
125.5 (C-1 " '), 110.0 (C-2 " '), 147.9 (C-3 " ') 149.4 (C-4 " '), 115.4 (C-5 " '), 123.4 (C-6 " '),
145.3 (C-7 " '), 114.5 (C-8 " '), 166.5 (C-9 " '), 55.7 (- OCH3)
Crude flavonoid powder 50g in 6 Example 2 of embodiment, is detached, eluant, eluent is dichloromethane with silica gel dry column chromatography
Full column is divided into 8 components by alkane-acetone (100: 0-1: 2) after the completion of elution, and each component elution can be obtained fraction 1-8.By each stream
Part carries out gradient elution by filler of ODS with mesolow flash chromatography, mobile phase for methanol-water (30: 70,50: 50,80: 20,
100: 0), wherein the elution of 50% methanol, 30% eluate are prepared into preparative HPLC, chromatographic condition is acetonitrile: water: formic acid (15:
85: 0.04-40: 60: 0.04), compound 16-22 can be obtained.Structure is as follows：
The hydrocarbon modal data of compound 16-22 is as follows：
Compound 16：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz,
DMSO-d6)δH：6.71 (1H, br s, H-6), 6.72 (1H, br s, H-8), 8.49 (2H, d, J=8.4Hz, H-2 ', 6 '),
7.18 (2H, d, J=8.4Hz, H-3 ', 5 '), 6.14 (1H, d, J=7.8Hz, H-1 "), 7.52 (2H, d, J=8.4Hz, H-
2 " ', 6 " '), 7.16 (2H, d, J=8.4Hz, H-3 " ', 5 " '), 7.84 (1H, d, J=15.6Hz, H-7 " '), 6.49 (1H, d, J
=15.6Hz, H-8 " ')13C-NMR (150MHz, DMSO-d6)δC：157.8 (C-2), 133.4 (C-3), 178.8 (C-4),
162.7 (C-5), 99.9 (C-6), 166.1 (C-7), 94.7 (C-8), 157.6 (C-9), 105.2 (C-10), 121.8 (C-1 '),
131.9 (C-2 ', 6 '), 116.1 (C-3 ', 5 '), 161.8 (C-4 '), 105.0 (C-1 "), 70.8 (C-2 "), 73.1 (C-3 "),
69.9 (C-4 "), 74.7 (C-5 "), 64.4 (C-6 "), 126.1 (C-1 " '), 130.7 (C-2 " ', 6 " '), 116.8 (C-3 " ',
5 " '), 161.4 (C-4 " '), 145.3 (C-7 " '), 114.8 (C-8 " '), 167.2 (C-9 " '),
Compound 17：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (400MHz,
CD3OD)δH：6.33 (1H, d, J=2.3Hz, H-6), 6.12 (1H, d, J=2.3Hz, H-8), 8.19 (2H, d, J=8.4Hz, H-
2 ', 6 '), 7.20 (2H, d, J=8.4Hz, H-3 ', 5 '), 5.24 (1H, d, J=7.2Hz, H-1 "), 7.16 (2H, s, H-2 " ',
6″′).13C-NMR (100MHz, CD3OD)δC：158.1 (C-2), 135.5 (C-3), 178.3 (C-4), 163.9 (C-5), 100.2
(C-6), 166.1 (C-7), 94.2 (C-8), 158.1 (C-9), 105.2 (C-10), 121.8 (C-1 '), 131.9 (C-2 ', 6 '),
116.1 (C-3 ', 5 '), 161.8 (C-4 '), 100.1 (C-1 "), 76.1 (C-2 "), 76.0 (C-3 "), 69.8 (C-4 "), 77.2
(C-5 "), 61.1 (C-6 "), 122.8 (C-1 " '), 112.0 (C-2 " ', 6 " '), 147.6 (C-3 " ', 5 " '), 137.8 (C-
4 " '), 171.2 (C-7 " ')
Compound 18：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molisch reactions are the positive.1H-NMR (600MHz,
DMSO-d6)δH：6.19 (1H, d, J=1.8Hz, H-6), 6.40 (1H, d, J=1.8Hz, H-8), 8.49 (2H, d, J=8.4Hz,
H-2 ', 6 '), 7.18 (2H, d, J=8.4Hz, H-3 ', 5 '), 5.36 (1H, d, J=7.8Hz, H-1 "), 6.87 (2H, s, H-
2 " ', 6 " ')13C-NMR (150MHz, DMSO-d6)δC：156.3 (C-2), 133.5 (C-3), 177.4 (C-4), 161.2 (C-
5), 98.7 (C-6), 164.2 (C-7), 93.6 (C-8), 156.4 (C-9), 103.9 (C-10), 121.8 (C-1 '), 131.9 (C-
2 ', 6 '), 116.1 (C-3 ', 5 '), 161.8 (C-4 '), 100.1 (C-1 "), 71.1 (C-2 "), 72.8 (C-3 "), 67.8 (C-
4 "), 72.4 (C-5 "), 62.0 (C-6 "), 119.1 (C-1 " '), 108.6 (C-2 " ', 6 " '), 145.5 (C-3 " ', 5 " '),
138.5 (C-4 " '), 165.4 (C=O)
Compound 19：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molisch reactions are the positive.1H-NMR (400MHz,
DMSO-d6)δH：6.23 (1H, d, J=2.5Hz, H-6), 6.37 (1H, d, J=2.5Hz, H-8), 8.52 (2H, d, J=8.3Hz,
H-2 ', 6 '), 7.21 (2H, d, J=8.4Hz, H-3 ', 5 '), 5.44 (1H, d, J=7.5Hz, H-1 "), 4.25 (d, J=12Hz,
Ha- 6 "), 4.32 (dd, J1=5Hz, J2=12Hz, Hb- 6 "), 6.71 (2H, d, J=8.7Hz, H-2 " ', H-6 " '), 7.56
(2H, d, J=8.7Hz, H-3 " ', H-5 " ')13C-NMR (100MHz, DMSO-d6)δC：157.2 (C-2), 133.5 (C-3),
177.9 (C-4, C=O), 161.8 (C-5), 99.6 (C-6), 165.4 (C-7), 94.6 (C-8), 157.7 (C-9), 104.5 (C-
10), 121.3 (C-1 '), 130.9 (C-2 ', 6 '), 115.6 (C-3 ', 5 '), 161.1 (C-4 '), 103.5 (C-1 "), 72.4 (C-
2 "), 74.1 (C-3 "), 69.5 (C-4 "), 74.8 (C-5 "), 64.0 (C-6 "), 120.1 (C-1 " '), 107.5 (C-2 " ', C-
6 " '), 142.5 (C-3 " ', C-5 " '), 136.62 (C-4 " '), 166.8 (C-7 " ')
Compound 20：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reacting positives.1H-NMR (400MHz, DMSO)
δH：6.20 (1H, d, J=2.2Hz, H-6), 6.44 (1H, d, J=2.2Hz, H-8), 6.60 (1H, d, J=8.1Hz, H-5 '),
7.49 (1H, d, J=3.0Hz, H-2 '), 7.12 (1H, dd, J1=8.1,3.0Hz, H-6 '), 5.28 (1H, d, J=7.8Hz, H-
1 "), 6.71 (2H, d, J=8.7Hz, H-2 " ', H-6 " '), 7.58 (2H, d, J=8.7Hz, H-3 " ', H-5 " ')13C-NMR
(100MHz, DMSO) δC：157.2 (C-2), 134.0 (C-3), 177.9 (C-4, C=O), 161.8 (C-5), 99.9 (C-6),
165.0 (C-7), 93.2 (C-8), 159.9 (C-9), 103.6 (C-10), 121.8 (C-1 '), 116.6 (C-2 '), 144.8 (C-
3 '), 148.2 (C-4 '), 115.2 (C-5 '), 121.7 (C-6 '), 103.3 (C-1 "), 73.1 (C-2 "), 74.1 (C-3 "),
69.0 (C-4 "), 75.2 (C-5 "), 63.9 (C-6 "), 161.9 (C-1 " '), 114.8 (C-2 " ', C-6 " '), 132.5 (C-3 " ',
C-5 " '), 121.1 (C-4 " '), 166.8 (C-7 " ')
Compound 21：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molisch reactions are the positive.1H-NMR (400MHz,
DMSO)δH：6.18 (1H, d, J=2.5Hz, H-6), 6.37 (1H, d, J=2.5Hz, H-8), 6.73 (1H, d, J=8.0Hz, H-
5 '), 7.45 (1H, d, J=3.0Hz, H-2 '), 7.59 (1H, dd, J1=8.0,3.0Hz, H-6 '), 5.44 (1H, d, J=
7.5Hz, H-1 "), 4.25 (1H, d, J=12.0Hz, Ha- 6 "), 4.32 (dd, J1=12.0,5.0Hb- 6 "), 3.3-3.8 (m,
Sugar protons), 6.90 (2H, s, H-2 " ', H-6 " ')13C-NMR (100MHz, DMSO) δC：156.3 (C-2), 133.2
(C-3), 177.0 (C-4, C=O), 161.2 (C-5), 98.9 (C-6), 165.0 (C-7), 93.6 (C-8), 160.9 (C-9),
103.6 (C-10), 121.7 (C-1 '), 115.6 (C-2 '), 144.8 (C-3 '), 148.8 (C-4 '), 115.2 (C-5 '), 120.7
(C-6 '), 104.3 (C-1 "), 73.9 (C-2 "), 74.1 (C-3 "), 69.3 (C-4 "), 76.2 (C-5 "), 62.9 (C-6 "),
119.1 (C-1 " '), 108.5 (C-2 " ', C-6 " '), 145.5 (C-3 " ', C-5 " '), 138.62 (C-4 " '), 165.8 (C-
7″′).
Compound 22：Yellow powder (methanol).Hydrochloric acid-magnesium powder, Molish reactions are the positive.1H-NMR (600MHz,
DMSO-d6)δH：6.13 (1H, d, J=2.0Hz, H-6), 6.38 (1H, d, J=2.0Hz, H-8), 8.01 (2H, d, J=8.5Hz,
H-2 ', 6 '), 6.87 (2H, d, J=8.5Hz, H-3 ', 5 '), 5.70 (1H, d, J=4.0Hz, H-1 "), 7.36 (2H, d, J=
8.5Hz H-2 " ', 6 " '), 6.79 (2H, d, J=8.5Hz, H-3 " ', 5 " '), 7.55 (1H, d, J=15.9Hz, H-7 " '),
6.15 (1H, d, J=15.9Hz, H-8 " '), 7.44 (2H, d, J=8.5Hz, H-2 " ", 6 " "), 6.77 (2H, d, J=8.5Hz,
H-3 " ", 5 " "), 7.61 (1H, d, J=15.8Hz, H-7 " "), 6.43 (1H, d, J=15.8Hz, H-8 " ") and13C-NMR
(150MHz, DMSO-d6)δC：157.1 (C-2), 133.1 (C-3), 177.5 (C-4), 161.1 (C-5), 98.5 (C-6), 164.5
(C-7), 94.0 (C-8), 156.9 (C-9), 104.3 (C-10), 121.0 (C-1 '), 131.1 (C-2 ', 6 '), 115.4 (C-3 ',
5 '), 160.1 (C-4 '), 98.5 (C-1 "), 73.1 (C-2 "), 73.5 (C-3 "), 70.1 (C-4 "), 74.2 (C-5 "), 62.3
(C-6 "), 125.5 (C-1 " '), 130.0 (C-2 " ', 6 " '), 115.2 (C-3 " ', 5 " '), 159.2 (C-4 " '), 144.1 (C-
7 " '), 113.3 (C-8 " '), 166.7 (C-9 " '), 125.1 (C-1 " "), 130.4 (C-2 " ", 6 " "), 115.5 (C-3 " ",
5 " "), 160.0 (C-4 " "), 145.5 (C-7 " ") and, 114.8 (C-8 " "), 165.2 (C-9 " ") and
Embodiment 7
The external anti-human lung carcinoma cell (H292) of 22 chromocor compounds and four kinds of Chinese chestnut crude flavonoid powders, gastric carcinoma cells
(N87), human liver cancer cell (HuH-7), the determination of activity of human pancreatic cancer cell (PANC-1) and human colon cancer cell (HT-29).
Sample is detected to the inhibited proliferation of tumor cell line by CCK-8 methods, obtains IC50。
Experimental implementation：
(1) cell culture
Exponential phase cell is collected, is counted, with complete medium again suspension cell, then adjusts cell concentration to conjunction
Suitable concentration is inoculated with 96 orifice plates, is inoculated with 100 μ l cell suspensions per hole, and last cell is at 37 DEG C, 100% relative humidity, 5%CO2Training
It supports and is incubated 24 hours in case.
(2) sample preparation
With DMSO diluted compounds to respective concentration.
(3)IC50Experiment
(a) exponential phase cell is collected, is counted, with complete medium again suspension cell, adjustment cell concentration to conjunction
Suitable concentration (being determined according to cell density optimization test result), is inoculated with 96 orifice plates, adds 100 μ l cell suspensions per hole.Cell is 37
DEG C, 100% relative humidity, 5%CO2It is incubated 24 hours in incubator.
(b) untested compound is diluted with DMSO, compound is from final concentration to 0 μM, totally 10 concentration points.
(c) after dosing, cell is placed in 37 DEG C, 100% relative humidity, 5%CO2It is incubated 72 hours in incubator；
(d) it inhales and abandons culture medium, complete medium of the addition containing 10%CCK-8, which is placed in 37 DEG C of incubators, to be incubated 2-6 hours.
(e) gently shake after in microplate reader measure 450nm wavelength at absorbance, using 650nm at absorbance as join
Than calculating inhibiting rate.
(4) inhibiting rate of the drug to growth of tumour cell is calculated as follows：
Growth of tumour cell inhibiting rate %=[(Ac-As)/(Ac-Ab)]× 100%
As：The OA (cell+CCK-8+ untested compounds) of sample
Ac：The OA (cell+CCK-8+DMSO) of negative control
Ab：The OA (culture medium+CCK-8+DMSO) of positive control
IC is fitted using software Graphpad Prism 550Curve simultaneously calculates IC50Value.
(5) Activity Results such as table l.
Table 1：The IC of 22 compounds and Chinese chestnut general flavone50Value.
Preferable in view of polyphenol chromocor compound and general flavone composition activity, the constituents and compound can be used as one kind or more
Kind can be used as the drug for the treatment of or auxiliary therapy cancer.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the principle of the present invention, it can also make several improvements and retouch, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (3)
1. polyphenol chromocor class compound shown in a kind of formula (I),
Wherein R1、R3Can simultaneously be H or OH or in which 1；R2、R4And R5It is glycosyl wherein at least to have 1, remaining is H or OH；
Glycosyl type has glucose (Glc), galactolipin (Gala), mannose (Man), rhamnose (Rham)；Shape can be also connected between sugar
At bioside or three glucosides, connection type has 1-2 connections, 1-3 connections, 1-4 connections to be connected with 1-6.The positions C-2 of the sugar, C-3
Position, C-4 and C-6 can also connect formula (II) or formula (III),
R in formula (II) and formula (III)6、R7And R8Can be H, OH, OCH simultaneously with identical meaning3Or H, OH, OCH3In
One or two；
The polyphenol chromocor class compound structure is as follows：
2. the preparation method of compound as described in claim 1, which is characterized in that include the following steps：
S21, medicinal material coarse powder is taken to utilize ultrasonic extraction, 70% ethyl alcohol measured using 6 times is extracted, and alcoholic solvent, residue is recovered under reduced pressure
It waves after being less than 5% to no alcohol taste or alcoholic degree, with water dissolution, obtains suspension；
S22, by gained suspension centrifuging and taking supernatant, supernatant exploitation polyamide column chromatography processing, with water-ethanol solution ladder
Degree elution, is enriched with 30%-70% alcohol elution fractions, obtains crude flavonoid powder；
S23, gained crude flavonoid powder is detached through silica gel dry column chromatography, eluant, eluent is dichloromethane-acetone, after the completion of elution
Full column is divided into 8 components, each component elution can be obtained into fraction 1-8, by each fraction mesolow flash chromatography using ODS as filler
Gradient elution is carried out, mobile phase is methanol-water, wherein the elution of 50% methanol, 30% eluate are prepared into preparative HPLC, chromatography
Condition is acetonitrile: water: formic acid obtains polyphenol flavone compound；
Wherein, the medicinal material coarse powder is one kind in Chinese chestnut involucre, chestnut flower, chestnut shell, Chinese chestnut Seed；
The mixed proportion of eluant dichloromethane-acetone is：100:0-1:2；
The mixed proportion of mobile phase methanol-water is：30:70,50:50,80:20,100:0；
Chromatographic condition acetonitrile: water: the mixed proportion of formic acid is：15:85:0.04-40:60:0.04.
3. the application of compound and thick general flavone that such as prepared by claim 2 in the drug for preparing antitumor cell growth,
It is characterized in that, the tumour is one kind in human lung cancer, human liver cancer, human pancreas cancer, human gastric cancer and human colon carcinoma.
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| WO2018028511A1 (en) * | 2016-08-12 | 2018-02-15 | 青岛海洋生物医药研究院股份有限公司 | Myricetin derivative and preparing method thereof, and application of same for treating colitis, preventing and treating colitis tumorigenesis, and treating colorectal cancer |
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