CN103342728A - Compound for preventing cancers and preparation method thereof - Google Patents
Compound for preventing cancers and preparation method thereof Download PDFInfo
- Publication number
- CN103342728A CN103342728A CN2013102677515A CN201310267751A CN103342728A CN 103342728 A CN103342728 A CN 103342728A CN 2013102677515 A CN2013102677515 A CN 2013102677515A CN 201310267751 A CN201310267751 A CN 201310267751A CN 103342728 A CN103342728 A CN 103342728A
- Authority
- CN
- China
- Prior art keywords
- compound
- prevention effect
- cancer prevention
- preparation
- obtains
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a compound for preventing cancers. The structure of the compound is shown in the formula II, wherein R represents Alpha-OH or Beta-OH. The compound has the quinone reductase inducing activity and can be used for preventing cancers. The invention further discloses a preparation method of the compound for preventing cancers. The method is simple, reliable, easy to implement, strong in operability and good in controllability.
Description
Technical field
The present invention relates to the biological chemistry field of medicaments, be specifically related to a kind of compound with cancer prevention effect that from Mao Ku Zhi, extracts and preparation method thereof.
Background technology
WITHANOLIDE type compound is the steroid compound that a class has the ergostane type of 28 carbon atoms, and its agent structure is shown in the formula III structure, and wherein, T in formula III structure part may be a kind of among a, b, c, d, e, f, g, the h.WITHANOLIDE type compound is divided into classic and non-classical type.Classical WITHANOLIDE type compound occupies absolute majority at nature, and these compounds mainly are distributed in the several genus such as Physalis (physalis) of Solanaceae (Solanaceae).That the biological activity of WITHANOLIDE type compound mainly shows is antitumor, anti-inflammatory, antibiotic and as the aspects such as antifeedant for insect of plant.Since the complex structure of this compounds, and chiral centre is a lot, and therefore, this compounds over-borrowing helps two dimensional NMR spectrum (2DNMR), circular dichroism spectrum (CD), X-single crystal diffraction modern techniquies such as (X-ray) is carried out structure and identified.
Mao Ku Zhi (Physalis angulata L.var.villosa Bonati) is the mutation of Solanaceae monkey flower Ku Zhi, be distributed in the province such as Zhejiang, Jiangxi, Hubei, Yunnan of Vietnam and China's Mainland more, generally be grown in the thick grass, do not have as yet at present and manually quote cultivation.Mao Ku Zhi, the nature and flavor bitter cold is returned lung channel, has clearing heat and detoxicating, relieve sore throat, reduces phlegm, effect such as diuresis, can be applicable to treatment of diseases such as pharyngalgia hoarsen, phlegm heat cough, dysuria, but up to now, very few to the report of Mao Ku Zhi both at home and abroad.
Whether have the effective constituent of quinone reductase induced activity in order to explore this plant with pharmaceutical use (being Mao Ku Zhi), the contriver studies the chemical ingredients that has the quinone reductase induced activity in the calyx extract of Mao Ku Zhi place and the chemical ingredients with cancer prevention effect.
Publication number is the immunosuppressor that the Chinese invention patent application of CN101422467A discloses a kind of WITHANOLIDE of containing type compound, and described immunosuppressor comprises compound or its optical isomer shown in following general formula (1) and/or the general formula (2);
Formula (1) formula (2)
Wherein, R1, R2, R3, R4, R5, R6, R7 are selected from H, OH, O-acyl, OMe, OEt, O
nPr, O
iPr, F, Cl, Br, NH
2, NO
2Or CN, wherein, 5,6 epoxies or two key, 14,27 ehter bonds connect or disconnection in the general formula (1), and 14,15 and 16,17 is epoxy and two keys in the general formula (2).
Summary of the invention
The invention provides a kind of compound with cancer prevention effect, have the quinone reductase induced activity, can be used for cancer prevention.
A kind of compound with cancer prevention effect is formula II structure;
Formula II;
Wherein, R is α-OH or β-OH.
This result with the proton nmr spectra of compound of cancer prevention effect and carbon-13 nmr spectra is as shown in table 2, sign by proton nmr spectra and carbon-13 nmr spectra, can show that the structure of the compound with cancer prevention effect that extracts is suc as formula shown in the II from Mao Ku Zhi, wherein, in the table 2, B is the formula II structure of R when being α-OH, and C is that R is the formula II structure of β-OH.
Table 2
(in?DMSO-d
6,B
1H?NMR500MHz,
13C?NMR125MHz;C
1H?NMR400MHz,
13C?NMR100MHz)
| B | C δ | H δ | C | C δ | H δ |
| 1 | 204.0 | ? | 1 | 200.8 | ? |
| 2 | 127.4 | 5.67dd(10.0，2.2) | 2 | 127.2 | 5.82dd(10.1，2.2) |
| 3 | 142.6 | 6.61ddd(10.0，4.8，2.2) | 3 | 143.2 | 6.79dd(10.1，5.0，2.2) |
| 4α | 35.1 | 3.08dt(19.8，2.2) | 4α | 37.0 | 3.44dt(20.4，2.2) |
| 4β | ? | 1.95dd(19.8，4.8) | 4β | ? | 2.46t(20.4) |
| 5 | 75.6 | ? | 5 | 75.3 | ? |
| 6 | 72.9 | 3.42t(2.2) | 6 | 72.9 | 3.82t(2.3) |
| 7α | 27.5 | 1.55td(11.6，2.4) | 7α | 27.6 | 1.77td(9.0，2.3) |
| 7β | ? | 1.72dd(11.6，3.1) | 7β | ? | 1.94dd(9.0，2.5) |
| 8 | 35.0 | 2.37m | 8 | 34.9 | 1.79t(8.9) |
| 9 | 34.5 | 1.69td(6.1，3.1) | 9 | 35.8 | 2.39m |
| 10 | 50.8 | ? | 10 | 52.1 | ? |
| 11α | 21.4 | 1.00dd(12.0，4.4) | 11α | 21.4 | 1.04dd(11.6，4.1) |
| 11β | ? | 2.06dd(12.0，2.9) | 11β | ? | 2.14dd(11.6，3.1) |
| 12α | 32.2 | 2.17dd(17.3，2.7) | 12α | 32.3 | 2.21dd(17.2，2.8) |
| 12β | ? | 1.63td(17.3，2.4) | 12β | ? | 1.62td(17.2，2.7) |
| 13 | 46.1 | ? | 13 | 46.1 | ? |
| 14 | 75.8 | ? | 14 | 82.9 | ? |
| 15 | 76.3 | 4.93s | 15 | 76.5 | 4.90s |
| 16 | 58.4 | 3.50s | 16 | 58.4 | 3.52s |
| 17 | 81.2 | ? | 17 | 80.7 | ? |
| 18 | 14.4 | 1.06s | 18 | 15.5 | 1.07s |
| 19 | 15.5 | 1.07s | 19 | 15.4 | 1.22 |
| 20 | 33.0 | 2.42t(6.4) | 20 | 33.3 | 2.41d(6.0) |
| 21 | 13.7 | 0.96d(7.0) | 21 | 13.8 | 0.95d |
| 22 | 76.2 | 4.32ddd(12.4，6.2，2.4) | 22 | 76.3 | 4.32m |
| 23α | 32.1 | 2.40dd(12.0，2.1) | 23α | 31.7 | 2.38dd(12.2，2.2) |
| 23β | ? | 1.63dd(12.0，2.4) | 23β | ? | 1.67dt(12.2，2.6) |
| 24 | 150.0 | ? | 24 | 149.9 | ? |
| 25 | 120.3 | ? | 25 | 120.3 | ? |
| 26 | 165.4 | ? | 26 | 165.4 | ? |
| 27 | 12.2 | 1.74s | 27 | 12.2 | 1.75s |
| 28 | 20.1 | 1.89s | 28 | 20.1 | 1.89s |
| OAc | 169.4 | ? | OAc | 169.2 | ? |
| ? | 20.8 | 2.01s | ? | 20.9 | 2.03s |
A kind of compound with quinone reductase induced activity has the quinone reductase induced activity, can be used for cancer prevention.
A kind of compound with quinone reductase induced activity is formula I structure;
Formula I.
This result with the proton nmr spectra of compound of quinone reductase induced activity and carbon-13 nmr spectra is as shown in table 1, by the sign of proton nmr spectra and carbon-13 nmr spectra, can show that the structure of the compound with quinone reductase induced activity that extracts is suc as formula shown in the I from Mao Ku Zhi.
Table 1
(in?DMSO-d
6,A
1H?NMR400MHz,
13C?NMR100MHz)
| A | C δ | H δ |
| 1 | 200.7 | ? |
| 2 | 126.5 | 5.90d(10.1) |
| 3 | 143.5 | 6.82ddd(10.1，4.8，2.4) |
| 4α | 31.6 | 2.42dd(12.0，2.4) |
| 4β | ? | 2.06td(12.0，2.4) |
| 5 | 76.0 | ? |
| 6 | 66.8 | 4.30m |
| 7α | 30.4 | 2.62d(4.2) |
| 7β | ? | 2.62d(4.2) |
| 8 | 39.5 | 2.04td(12.5，3.1) |
| 9 | 39.3 | 1.82td(12.5，3.5) |
| 10 | 55.9 | ? |
| 11α | 22.0 | 1.13t(14.0) |
| 11β | ? | 2.14d(14.0) |
| 12α | 37.5 | 1.68dd(13.1，4.3) |
| 12β | ? | 1.16d(13.1) |
| 13 | 52.1 | ? |
| 14 | 80.0 | ? |
| 15 | 83.0 | 5.10d(2.1) |
| 16 | 121.3 | 5.60d(2.5) |
| 17 | 160.8 | ? |
| 18 | 15.6 | 0.93s |
| 19 | 7.4 | 1.01s |
| 20 | 34.1 | 2.43dd(11.0，3.8) |
| 21 | 17.4 | 1.05d(7.1) |
| 22 | 78.0 | 4.30m |
| 23α | 33.1 | 2.43dd(12.0，3.8) |
| 23β | ? | 1.70dd(12.0，4.3) |
| 24 | 150.2 | ? |
| 25 | 120.3 | ? |
| 26 | 165.6 | ? |
| 27 | 12.2 | 1.74s |
| 28 | 20.1 | 1.88s |
| OAc | 169.2 | ? |
| ? | 21.0 | 1.94s |
The present invention also provides a kind of to have the compound of quinone reductase induced activity and has the preparation method of the compound of cancer prevention effect, and simple and reliable, easy to implement, workable, controllability is good.
Described have the compound of quinone reductase induced activity and have the preparation method of the compound of cancer prevention effect, may further comprise the steps:
1) Mao Ku Zhi place calyx is extracted with extracting solvent refluxing, obtain extracting solution;
Described extraction solvent is alcohol or the aqueous solution that contains the alcohol of pure percent by volume 50%～99.99%;
2) extracting solution that obtains in the step 1) is concentrated, use organic solvent extraction behind the dilute with water, obtain extraction liquid;
3) with step 2) in the extraction liquid that obtains cross chromatographic column through concentrating the back, through concentrating the blending ingredients that obtains containing the compound with quinone reductase induced activity and have the compound of cancer prevention effect behind the first eluent system wash-out;
Described first eluent system is the eluting solvent of being made up of sherwood oil and ethyl acetate;
4) blending ingredients that obtains in the step 3) is crossed the preparative liquid chromatography post, through concentrating the compound that obtains having the compound of quinone reductase induced activity and have the cancer prevention effect behind the second eluent system wash-out;
Described second eluent system is the eluting solvent of being made up of the first alcohol and water.
In order to obtain better invention effect, below as of the present invention preferred:
In the step 1), described alcohol is methyl alcohol or ethanol, selects these two kinds when pure, and the extraction efficiency of extracting solvent is higher.More preferably, described extraction solvent is methyl alcohol or the aqueous ethanolic solution that contains ethanol percent by volume 95%, the character of this extraction solvent is very suitable for extracting Mao Ku Zhi place calyx, and the classical WITHANOLIDE type that can more fully obtain among the Mao Ku Zhi has the compound of quinone reductase induced activity and the compound that classical WITHANOLIDE type has the cancer prevention effect.
Each extraction solvent that adds with respect to 3～5 times of weight of Mao Ku Zhi place calyx weight, 90 ℃～100 ℃ refluxing extraction, each time of extracting is 2h～3h, extracts 2～4 times.Under such extraction conditions, not only saved the usage quantity of Mao Ku Zhi place calyx and extraction solvent, and the compound that has the quinone reductase induced activity in the calyx of Mao Ku Zhi place fully can have been extracted with the compound with cancer prevention effect, improved extraction yield.
Step 2) in, described organic solvent is ethyl acetate, according to the similar principle that mixes, adopt ethyl acetate extraction, make the compound that has the compound of quinone reductase induced activity among the Mao Ku Zhi and have a cancer prevention effect almost all concentrate in the acetic acid ethyl acetate extract, be conducive to extract high purity more and more have the compound of quinone reductase induced activity and have the compound of cancer prevention effect.
In the step 3), the used filler of described chromatographic column is aluminum oxide or 100～200 purpose column layer chromatography silicone rubbers.The medicinal extract that described extraction liquid obtains after concentrating and the weight ratio of filler are 1:10～20, more preferably 1:15～20.The volume ratio of the described first eluent system PetroChina Company Limited. ether and ethyl acetate is 5～15:1, more preferably 10:1.
In the step 4), the used filler of described preparative liquid chromatography post is the C-18 column layer chromatography silicone rubber, be specially ODS(octadecyl silane) the C-18 column layer chromatography silicone rubber, be to be the C18 filler of matrix bonding with silica gel, octadecyl silane is octadecyl silane.Described second eluent system is the methanol aqueous solution that contains methyl alcohol percent by volume 60%～80%, the methanol aqueous solution that more preferably contains methyl alcohol percent by volume 65%～75% is by concentrating the compound that obtains having the compound of quinone reductase induced activity respectively and have the cancer prevention effect behind the isocratic elution.
Phase II metabolic enzyme is significant for the generation of prophylaxis of tumours, quinone reductase (NAD (P) H:quinone reductase as one of phase II metabolic enzyme, QR) be that a class has detoxifcation, remove the cytoprotective enzyme of free radical effect, the composition that quinone reductase is had induced activity is considered to have the cancer prevention effect at the tumour initial period, the rise of quinone reductase level is considered to the important mechanisms (reference: Su BN that preventing cancer takes place, Gu JQ, Kang YH, Park EJ, Pezzuto JM, Kinghorn AD.Induction of the phase II enzyme, Quinone Reductase, by withanolides and norwithanolides from solanaceous species.Mini-Rev Org Chem2004; 1:115-123.Rosana I.Misico, Lynda L.Song, Adriana S.Veleiro, et.al.Induction of Quinone Reductase by Withanolides.J.Nat.Prod.2002; 65:677-680.).Carry out the chemoprophylactic screening of cancer with mtt assay, be that quinone reductase is induced experiment, the result shows that the present invention has the compound of quinone reductase induced activity and has the compound of cancer prevention effect, all can induce the expression of quinone reductase, have the cancer prevention effect.The present invention has the compound of quinone reductase induced activity can use the medicine for preparing cancer prevention with the compound with cancer prevention effect.
Compared with prior art, the present invention has following advantage:
The present invention has the compound of quinone reductase induced activity and has compound content height in Mao Ku Zhi plant of cancer prevention effect, and production cost is lower.The compound that the present invention has the compound of quinone reductase induced activity and has the cancer prevention effect also has cytotoxic activity to cancer cells except the expression that can induce quinone reductase.Therefore, have the compound of quinone reductase induced activity and have the compound of cancer prevention effect can be for the preparation of the medicine of prophylaxis of tumours in the present invention.
The present invention has the compound of quinone reductase induced activity and has the preparation method of the compound of cancer prevention effect, from Mao Ku Zhi plant, separate to obtain the compound monomer that the WITHANOLIDE type has the quinone reductase induced activity with some common cheap reagent such as alcohol, ethyl acetate and silica gel, reduced production cost.This preparation method is simple and reliable, and is workable, and the efficient height can carry out industrialized production, is conducive to apply, and has a good application prospect.
Description of drawings
Fig. 1 is the proton nmr spectra of the compd A with quinone reductase induced activity of embodiment 1 preparation;
Fig. 2 is the carbon-13 nmr spectra of the compd A with quinone reductase induced activity of embodiment 1 preparation;
Fig. 3 is the proton nmr spectra of the compd B with cancer prevention effect of embodiment 1 preparation;
Fig. 4 is the carbon-13 nmr spectra of the compd B with cancer prevention effect of embodiment 1 preparation;
Fig. 5 is the proton nmr spectra of the Compound C with cancer prevention effect of embodiment 1 preparation;
Fig. 6 is the carbon-13 nmr spectra of the Compound C with cancer prevention effect of embodiment 1 preparation.
Embodiment
By the following examples content of the present invention is described in further detail, but embodiment should be interpreted as limitation of the present invention.Do not breaking away under the above-mentioned state of mind of the present invention, various replacement means or change according to ordinary skill knowledge and conventional means are made all are included within the protection domain of the present invention.
Embodiment 1
1) 1Kg Mao Ku Zhi place calyx is packed into extractor, each aqueous ethanolic solution that contains ethanol percent by volume 95% (being aqueous ethanolic solution 5Kg) that adds with respect to 5 times of Mao Ku Zhi place calyx weight, 90 ℃ of refluxing extraction, each time of extracting is 2h, extract 4 times (aqueous ethanolic solution amounts to 20Kg) united extraction liquid altogether;
2) extracting solution that obtains in the step 1) is concentrated (recovery ethanol) and do near, obtain thick medicinal extract 130g, after the dilution of 1L water, use the 1L ethyl acetate extraction, coextraction 3 times, combining extraction liquid at every turn;
3) with step 2) in the extraction liquid that obtains concentrate the medicinal extract 20g that obtains through silica gel chromatographic column (wet method dress post, sample on the dry method), wherein, the used filler of silica gel chromatographic column is 100 purpose column layer chromatography silicone rubbers, the weight ratio of medicinal extract and column layer chromatography silicone rubber is 1:15, and through being the sherwood oil of 10:1 and the eluting solvent wash-out that ethyl acetate is formed by volume ratio, this eluting solvent uses 4L at every turn again, co-elute 3 times merges elutriant;
4) elutriant that obtains in the step 3) is crossed the preparative liquid chromatography post after concentrating, wherein, the used filler of chromatographic column is ODS C-18 column layer chromatography silicone rubber, be 65% methanol aqueous solution wash-out through containing the methyl alcohol percent by volume again, obtain having the compd A (C of quinone reductase induced activity after concentrating respectively
30H
39ClO
7) 14mg, name PA-14J
2, i.e. formula I structure; Compd B (C with cancer prevention effect
30H
42O
10) 16mg, name PA-11K
1, namely in the formula II structure, R is α-OH; Compound C (C with cancer prevention effect
30H
42O
10) 20mg, name PA-19I
2, namely in the formula II structure, R is β-OH;
Formula I; Formula II.
The proton nmr spectra of the compd A with quinone reductase induced activity of embodiment 1 preparation as shown in Figure 1, the carbon-13 nmr spectra of the compd A with quinone reductase induced activity of embodiment 1 preparation as shown in Figure 2, the proton nmr spectra of the compd B with cancer prevention effect of embodiment 1 preparation as shown in Figure 3, the carbon-13 nmr spectra of the compd B with cancer prevention effect of embodiment 1 preparation as shown in Figure 4, the proton nmr spectra of the Compound C with cancer prevention effect of embodiment 1 preparation as shown in Figure 5, the carbon-13 nmr spectra of the Compound C with cancer prevention effect of embodiment 1 preparation is as shown in Figure 6.
Can characterize by Fig. 1 and Fig. 2, the compd A structure with quinone reductase induced activity of embodiment 1 preparation is suc as formula shown in the I.
Can characterize by Fig. 3 and Fig. 4, the compd B structure with cancer prevention effect of embodiment 1 preparation is suc as formula shown in the II, and R is α-OH in the formula II structure.
Can characterize by Fig. 5 and Fig. 6, the Compound C structure with cancer prevention effect of embodiment 1 preparation is suc as formula shown in the II, and R is β-OH in the formula II structure.
1) 1Kg Mao Ku Zhi place calyx is packed into extractor, each aqueous ethanolic solution that contains ethanol percent by volume 95% (being aqueous ethanolic solution 3Kg) that adds with respect to 3 times of Mao Ku Zhi place calyx weight, 100 ℃ of refluxing extraction, each time of extracting is 3h, extract 2 times (aqueous ethanolic solution amounts to 6Kg) united extraction liquid altogether;
2) extracting solution that obtains in the step 1) is concentrated (recovery ethanol) and do near, obtain thick medicinal extract 110g, after the dilution of 1L water, use the 1L ethyl acetate extraction, coextraction 3 times, combining extraction liquid at every turn;
3) with step 2) in the extraction liquid that obtains concentrate the medicinal extract 16g that obtains through silica gel chromatographic column (wet method dress post, sample on the dry method), wherein, the used filler of silica gel chromatographic column is 200 purpose column layer chromatography silicone rubbers, the weight ratio of medicinal extract and column layer chromatography silicone rubber is 1:20, and through being the sherwood oil of 10:1 and the eluting solvent wash-out that ethyl acetate is formed by volume ratio, this eluting solvent uses 4L at every turn again, co-elute 3 times merges elutriant;
4) elutriant that obtains in the step 3) is crossed the preparative liquid chromatography post after concentrating, wherein, the used filler of chromatographic column is ODS C-18 column layer chromatography silicone rubber, be 75% methanol aqueous solution wash-out through containing the methyl alcohol percent by volume again, obtain having the compd A (C of quinone reductase induced activity after concentrating respectively
30H
39ClO
7) 11mg, name PA-14J
2, i.e. formula I structure; Compd B (C with cancer prevention effect
30H
42O
10) 12mg, name PA-11K
1, namely in the formula II structure, R is α-OH; Compound C (C with cancer prevention effect
30H
42O
10) 15mg, name PA-19I
2, namely in the formula II structure, R is β-OH.
The compd A with quinone reductase induced activity of preparation and have the compd B of cancer prevention effect and C characterizes by proton nmr spectra and carbon-13 nmr spectra among the embodiment 2 has above-mentioned structure.
Embodiment 3(quinone reductase is induced experiment)
The take the logarithm rat liver cancer cell (Hepa1c1c7 purchases in ATCC) of phase, every hole 1 * 10
4Be inoculated on 96 orifice plates, every hole adds the DMEM substratum of 200 μ L, behind the 12h, abandon supernatant, the compd A with quinone reductase activity (0.106mg) that in 3 multiple holes, adds embodiment 1 preparation then, select 3 else and add the compd B with cancer prevention effect in the holes again, adding has the Compound C of cancer prevention effect in 3 other multiple holes at last, corresponds to compound dosing group A, compound dosing group B and compound dosing group C respectively; In 3 multiple holes, add 4 '-bromine flavones (0.106mg), positive contrast dosing group; In 3 multiple holes, add dimethyl sulfoxide (DMSO) (0.106mg), negative control group; Blank group is the DMEM substratum of 200 μ L of not dosing.After cultivating 24h, abandon supernatant liquor.Add digitonin (10 μ g) in compound dosing group (A, B, C), positive control dosing group, negative control group and the blank group respectively and make lysis, the solution 200 μ L that add band MTT again cultivate 5min, the solution of MTT is by MTT (tetrazolium bromide, green skies reagent company) is dissolved in phosphoric acid buffer (PBS, pH=7.3) form in, MTT concentration is 0.5mg/mL, and the 550nm place surveys the IR value on microplate reader.(induce multiple; IR=[(dosing group OD value-blank group OD value)/(negative control group OD value-blank group OD value)]/(1-proliferation inhibition rate %); Wherein, the blank group of proliferation inhibition rate %=(OD value-dosing group OD value)/blank group OD value.）
The result shows, the compd B with cancer prevention effect of the compd A with quinone reductase induced activity of embodiment 1 preparation and embodiment 1 preparation and C IR value under 1mM (0.106mg) administration final concentration be respectively 1.51,1.79 and 2.85(positive control drug 4 '-the IR value of bromine flavones is 2.46); The multiple of IR value for inducing with respect to the quinone reductase of negative control DMSO, the result shows that compound can induce the expression of quinone reductase, can think to have the cancer prevention effect at the tumour initial period.Described cell is all purchased in ATCC.
Table 1
| Group | The IR value |
| Compound dosing group A | 1.51±0.05 |
| Compound dosing group B | 1.79±0.01 |
| Compound dosing group C | 2.85±0.02 |
| Blank group | 1.00±0.09 |
| Positive control dosing group | 2.46±0.07 |
Experimental result shows, the compd B with cancer prevention effect of the compd A with quinone reductase induced activity of embodiment 1 preparation and embodiment 1 preparation and C have quinone reductase induced activity in various degree under 1mM (0.106mg) administration final concentration, wherein, have the cancer prevention effect Compound C the IR value greater than positive control drug 4 '-the bromine flavones, show that the Compound C with cancer prevention effect has good quinone reductase induced activity, has cancer prevention effect preferably at the tumour initial period.
The experiment of embodiment 4(cytotoxic activity)
The take the logarithm every hole 1 * 10 of cell of phase
4Be inoculated on 96 orifice plates, every hole adds the DMEM substratum of 200 μ L, behind the 12h, abandon supernatant liquor, in multiple hole, add the compd A with quinone reductase induced activity of embodiment 1 preparation and the compd B with cancer prevention effect and the C of embodiment 1 preparation then, respectively by 0,2.5,5,10,20,30,3 multiple holes are established in 50 μ M concentration administrations, each concentration, are compound administration group A, compound administration group B and compound administration group C; After cultivating 24h, abandon supernatant liquor.In compound administration group A, compound administration group B, compound administration group C, add the solution 50 μ L cultivation 4h that is with MTT respectively, the solution of MTT is by MTT (tetrazolium bromide, green skies reagent company) is dissolved in phosphoric acid buffer (PBS, pH=7.3) form in, MTT concentration is 0.5mg/mL, add 100 μ L dimethyl sulfoxide (DMSO) (DMSO) more respectively to each hole, vibration 1h, the 570nm place surveys OD(optical density(OD) optical density on microplate reader) value.(proliferation inhibition rate that the gradient dosing dosage of each strain tumor cell line is corresponding with it carries out The Fitting Calculation, obtains Nonlinear regression equation, IC
50Value refers to that the tumor cell line proliferation inhibition rate is 50% o'clock dosing dosage.Wherein, the blank group of proliferation inhibition rate %=(OD value-administration group OD value)/blank group OD value.) result's demonstration, after the dosing, activity of tumor cells obviously descends.This experiment is carried out the detection of cytotoxic activity to human lung carcinoma cell Calu-1, H1975, H460 respectively, and above-mentioned cell strain is all purchased in ATCC, and concrete result is as shown in table 2.
Table 2
Experimental result shows, the compd B with cancer prevention effect of the compd A with quinone reductase induced activity of embodiment 1 preparation and embodiment 1 preparation and C have in various degree restraining effect to different tumour cells, all can play certain cancer prevention effect.Wherein, have the cancer prevention effect compd B cytotoxic activity relatively a little less than, the compd A with quinone reductase induced activity has medium cytotoxic activity, has the IC of the Compound C of cancer prevention effect
50Value all below 14 μ M, shows that it has the good cell cytotoxic activity, can play cancer prevention effect preferably.
Claims (9)
1. the compound with cancer prevention effect is characterized in that, is formula II structure;
Formula II;
Wherein, R is α-OH or β-OH.
2. the preparation method with compound of cancer prevention effect according to claim 1 is characterized in that, may further comprise the steps:
1) Mao Ku Zhi place calyx is extracted with extracting solvent refluxing, obtain extracting solution;
Described extraction solvent is alcohol or the aqueous solution that contains the alcohol of pure percent by volume 50%～99.99%;
2) extracting solution that obtains in the step 1) is concentrated, use organic solvent extraction behind the dilute with water, obtain extraction liquid;
3) with step 2) in the extraction liquid that obtains cross chromatographic column through concentrating the back, through concentrating the blending ingredients that obtains containing the compound with quinone reductase induced activity and have the compound of cancer prevention effect behind the first eluent system wash-out;
Described first eluent system is the eluting solvent of being made up of sherwood oil and ethyl acetate;
4) blending ingredients that obtains in the step 3) is crossed the preparative liquid chromatography post, through concentrating the compound that obtains having the cancer prevention effect behind the second eluent system wash-out;
Described second eluent system is the eluting solvent of being made up of the first alcohol and water.
3. the preparation method with compound of cancer prevention effect according to claim 2 is characterized in that, in the step 1), described alcohol is methyl alcohol or ethanol.
4. the preparation method with compound of cancer prevention effect according to claim 2, it is characterized in that, in the step 1), each extraction solvent that adds with respect to 3～5 times of weight of Mao Ku Zhi place calyx weight, 90 ℃～100 ℃ refluxing extraction, each time of extracting is 2h～3h, extracts 2～4 times.
5. the preparation method with compound of cancer prevention effect according to claim 2 is characterized in that step 2) in, described organic solvent is ethyl acetate.
6. the preparation method with compound of cancer prevention effect according to claim 2, it is characterized in that, in the step 3), the used filler of described chromatographic column is aluminum oxide or 100～200 purpose column layer chromatography silicone rubbers, and the medicinal extract that described extraction liquid obtains after concentrating and the weight ratio of filler are 1:10～20.
7. the preparation method with compound of cancer prevention effect according to claim 2 is characterized in that, in the step 3), the volume ratio of the described first eluent system PetroChina Company Limited. ether and ethyl acetate is 5～15:1.
8. the preparation method with compound of cancer prevention effect according to claim 2 is characterized in that, in the step 4), the used filler of described preparative liquid chromatography post is the C-18 column layer chromatography silicone rubber.
9. the preparation method with compound of cancer prevention effect according to claim 2 is characterized in that, in the step 4), described second eluent system is the methanol aqueous solution that contains methyl alcohol percent by volume 60%～80%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310267751.5A CN103342728B (en) | 2013-06-28 | 2013-06-28 | Compound for preventing cancers and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310267751.5A CN103342728B (en) | 2013-06-28 | 2013-06-28 | Compound for preventing cancers and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103342728A true CN103342728A (en) | 2013-10-09 |
| CN103342728B CN103342728B (en) | 2015-05-20 |
Family
ID=49277563
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310267751.5A Active CN103342728B (en) | 2013-06-28 | 2013-06-28 | Compound for preventing cancers and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103342728B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103961369A (en) * | 2014-05-16 | 2014-08-06 | 富阳科兴生物化工有限公司 | Application of physagulin P in preparation of antitumor drug |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101422467A (en) * | 2007-11-02 | 2009-05-06 | 华东理工大学 | Immunosuppressant containing Withanolide type compound |
-
2013
- 2013-06-28 CN CN201310267751.5A patent/CN103342728B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101422467A (en) * | 2007-11-02 | 2009-05-06 | 华东理工大学 | Immunosuppressant containing Withanolide type compound |
Non-Patent Citations (1)
| Title |
|---|
| QING-PING HE ET AL: "Cytotoxic Withanolides from Physalis angulata L.", 《CHEMISTRY & BIODIVERSITY》, vol. 4, 31 December 2007 (2007-12-31), pages 443 - 449 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103961369A (en) * | 2014-05-16 | 2014-08-06 | 富阳科兴生物化工有限公司 | Application of physagulin P in preparation of antitumor drug |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103342728B (en) | 2015-05-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101240005A (en) | Method for preparing platycodin D from balloon-flower root and application thereof in anti-cancer medicament | |
| CN102940687B (en) | A kind of Fructus Toosendan extract and uses thereof | |
| CN105503786B (en) | Split Lignanoids compounds-smartweed diffractive ring lignan glucoside E and preparation method thereof | |
| CN108003214A (en) | A kind of saponin compound and its methods and applications extracted from the rhizoma bolbostemmae | |
| CN102516344B (en) | Compound with antitumor activity and preparation method and application thereof | |
| CN103333217B (en) | Compound having quinone reductase induced activity and preparation method thereof | |
| CN104586945A (en) | Hop total flavonoid extract as well as preparation method and application thereof in preparing medicines for preventing and treating liver injury and cancers | |
| Zhan et al. | Flavans with potential anti-inflammatory activities from Zephyranthes candida | |
| CN103342728B (en) | Compound for preventing cancers and preparation method thereof | |
| CN104774232B (en) | Hemsleya amabilis cucurbitane tetracyclotriterpene compounds, pharmaceutical composition containing compounds and application thereof | |
| CN110922440A (en) | Indole nitrogen glycoside compound, extraction method and application in preparation of medicine for preventing and treating nervous system diseases | |
| CN105949266A (en) | Withana lactide compound, method for extracting same and application of withana lactide compound | |
| CN107880084B (en) | Method for extracting high-purity verbascoside from campsis plant flowers | |
| CN107033156B (en) | Aporphine alkaloid Laurodionine B and its preparation method and application | |
| CN109879921A (en) | Compound with anti-tumor activity separated from rhizoma anemarrhenae and preparation method thereof | |
| CN101856382B (en) | Novel method for extracting and separating platycodin effective ingredients | |
| CN102675252B (en) | There is Cesong alkyl type diterpine compound and the application thereof of anti-tumor activity | |
| CN106008641A (en) | Withania somnifera lactide compound, method for extracting same and application of withania somnifera lactide compound | |
| CN102775461A (en) | Method for preparing 20 (R)-ginseniside Rg3 | |
| CN102379973B (en) | Preparation method and application of effective part of solanaceae physalis plant | |
| CN101851271A (en) | Glaucocalyxin D derivative, preparation method and application thereof | |
| CN106243184B (en) | Extraction separation method and the application of triterpenoid and the compound drop in a kind of chinaberry bitter principle class four | |
| CN104059950B (en) | A kind of preparation method of compound | |
| CN108456136B (en) | Extraction and separation method and application of pimaric dienoic acid | |
| CN113582828B (en) | Natural compound for resisting liver cancer, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |