CN103288911A - Bufalin glycosylation derivative and preparation method and application thereof in preparation of anti-tumour medicaments - Google Patents
Bufalin glycosylation derivative and preparation method and application thereof in preparation of anti-tumour medicaments Download PDFInfo
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Abstract
The invention discloses a bufalin glycosylation derivative and a preparation method and application thereof in preparation of anti-tumour medicaments. The structure of the bufalin glycosylation derivative is shown in a formula I or formula II. The synthesis method of the bufalin glycosylation derivative comprises the following steps of: oxidizing bufalin into a ketone-form derivative by pyridinium chloropyridine hydrochloride; then performing a nucleophilic addition reaction with methoxyamine hydrochloride; removing monomolecular water to generate an oxime-form intermediate; enabling the oxime-form intermediate to react with a tert-butylamine borane hydrochloride compound to generate alpha-configuration aglycone and beta-configuration aglycone; and finally enabling the aglycones of two configurations to react with reducing sugar respectively to generate the bufalin glycosylation derivative. According to the method disclosed by the invention, bufalin is modified to be the glycosylation derivative thereof, and the water solubility can be improved while the anti-tumour activity is kept and enhanced; and moreover, in-vivo experiments prove that the toxic and side effects of the bufalin glycosylation derivative disclosed by the invention on normal cells are reduced compared with those of a parent compound of bufalin.
Description
Technical field
The invention belongs to the pharmaceutical chemistry field, be specifically related to Toadpoison Medicine glycosylated derivative and method for making thereof and the purposes in the preparation antitumor drug.
Background technology
Malignant tumour is the major disease of harm humans health always.According to WHO statistics, the newly-increased case of malignant tumour in whole world every year is about 1,000 ten thousand, dead about 8,000,000 examples.In China, malignant tumour accounts for first of city resident's mortality ratio gradually.At present, the main methods for the treatment of of malignant tumour comprises operation, radiation and chemotherapy.Wherein chemotherapy still plays an important role, but that the most of chemotherapeutics that uses clinically has toxicity more is big, and security is low and easily produce shortcoming such as resistance.The antitumor drug of research high-efficiency low-toxicity is the focus of whole world the world of medicine always.
Natural product is the important source of antitumor drug.Antitumor drugs such as taxol, vinealeucoblastine(VLB), camptothecine are all from natural product.In addition, some natural product toxicity are big, poorly water-soluble, but through behind the structural modification, also become the antitumor drug of clinical use, for example Etoposide and teniposide.Therefore natural constituent is carried out the important channel that structure of modification also is the antitumor drug research and development.
The dried venom of toads is that the white juice of bufo gargarizans Cantor (Bufo bufo gargarizans Cantor) or Bufo melanostictus (Bufo melanostictus Schneider) ear rear gland and skin gland secretion processes (" Chinese pharmacopoeia).Contain B-mode cardenolide in the dried venom of toads.This constituents not only has good cardiac activity, also has very strong anti-tumor activity.Toadpoison Medicine is the main active ingredient in the dried venom of toads.It is reported, Toadpoison Medicine has its propagation of inhibition and induces its effect of apoptosis [Tian Xin human leukemia cell HL60, gastric carcinoma cells MGc-803, human liver cancer cell SMMC7721 etc., Luo Ying, Liu Yunpeng, Deng. Toadpoison Medicine is induced the influence of in the HL-60 apoptosis process Bcl-2, Survivin, Smac/DIABLO being expressed. Chinese Journal of Hematology, 2006, (01): 21-24; Chen Xiaoyi, Xu Ruicheng, Chen Li, etc. Toadpoison Medicine is to the cytotoxicity of SGC-7901 MGc-803. herbal medicine, 2000, (12): 920-922; Chen Xiaoyi exhales literary composition bright, Xu Ruicheng, etc. the influence that Toadpoison Medicine is expressed cytotoxicity and the growth related gene of liver cancer cell SMMC7721. Chinese J Pharmacol Toxicol, 2001, (04): 293-296.].Study on mechanism shows that the antitumor action of Toadpoison Medicine may influence apoptosis relevant gene (bcl-2, c-myc and Tiam1) relevant (Y.Masuda, N et al.Leuk.Res.1995,19,549 – 556 with it; K.Nobuko, et al.Oncogene1999,18:2413 – 2421.).
Though Toadpoison Medicine has remarkable antitumor effect, it is suitable with Normocellular toxicity to tumour cell, non-selectivity, and its poorly water-soluble.
Summary of the invention
In order to overcome the defective of Toadpoison Medicine poor selectivity (suitable with Normocellular toxicity to tumour cell) and poorly water-soluble, primary and foremost purpose of the present invention is to introduce in the 3-position of Toadpoison Medicine to replace glycosyl, and a kind of Toadpoison Medicine glycosylated derivative with anti-tumor activity is provided.Toadpoison Medicine glycosylated derivative of the present invention has significantly improved water-soluble, still has very strong anti-tumor activity simultaneously, and Normocellular toxicity is obviously reduced.
The preparation method of the Toadpoison Medicine glycosylated derivative that another object of the present invention is to provide above-mentioned.
The purposes of the Toadpoison Medicine glycosylated derivative that a further object of the present invention is to provide above-mentioned in the preparation antitumor drug.
Purpose of the present invention is achieved through the following technical solutions:
A kind of Toadpoison Medicine glycosylated derivative, its structure is suc as formula shown in I or the formula II:
Wherein R is the reducing sugar glycosyl, is preferably D-glucose sugar (D-Glu), L-glucose sugar (L-Glu), Fucose (Fuc), wood sugar (Xyl), lyxose (Lxy) or Arabic glycosyl (Ara).
The synthetic method of above-mentioned Toadpoison Medicine glycosylated derivative may further comprise the steps:
(1) Toadpoison Medicine is mixed with mol ratio 1:2 with oxygenant, be dissolved in the organic solvent, after the reaction, silica gel column chromatography separates under the room temperature, obtains the intermediate B ufalinone of 3 oxidations of Toadpoison Medicine;
(2) oxidation products Bufalinone is mixed with mol ratio 1:2.5 with the organic amine salt hydrochlorate, be dissolved in the organic solvent, add alkaline reagents, react under the room temperature condition, obtain oxime formula intermediate;
(3) reductive agent of 3 times of oxime formula intermediate molar weights of adding in oxime formula intermediate is used organic solvent dissolution then, reacts under the ice-water bath condition, and silica gel column chromatography separates, and obtains the aglycon isomer of 3 α of Toadpoison Medicine and two kinds of configurations of β;
(4) the two kinds of aglycon isomer and the reducing sugar that respectively step (3) are obtained place organic solvent to react, and separate the Toadpoison Medicine glycosylated derivative that obtains suc as formula structure shown in I or the formula II through high performance liquid phase;
The preferred pyridinium chlorochromate hydrochloride of the described oxygenant of step (1) (PCC), the preferred CH of organic solvent
2Cl
2, preferred 2.5h of reaction times;
The described silica gel column chromatography of step (1) separates, and is the cyclohexane/ethyl acetate mixing solutions gradient elution with volume ratio 1:1;
The preferred methoxamine hydrochloride of the described organic amine salt hydrochlorate of step (2), organic solvent is methyl alcohol, and alkaline reagents is pyridine, and the reaction times is 2.5-3h;
The described reductive agent preferred tertiary of step (3) butylamine base borane hydrochloride mixture, organic solvent is that volume ratio is dioxane/alcohol mixeding liquid of 2:1, reaction times 3.5h;
The described silica gel column chromatography of step (3) separates, and is the cyclohexane/ethyl acetate mixing solutions gradient elution with volume ratio 4:1;
The described reducing sugar of step (4) is glucose, Fucose, wood sugar, lyxose or pectinose;
The described organic solvent of step (4) is that volume ratio is dimethyl formamide/alcohol mixeding liquid of 3:1, and the reaction times is 48h;
The described high performance liquid phase of step (4) separates, specifically: performance liquid chromatographic column (waters, C18,5 μ m, 20 * 250mm), flow velocity 8ml/min detects wavelength 296nm, the acetonitrile-water of moving phase 45%.
The compound of gained among this preparation method, by UV spectrum, infrared spectra, mass spectrum and magnetic nuclear resonance method carry out structure to be identified, and carries out purity detecting with the HPLC method.
Toadpoison Medicine glycosylated derivative of the present invention and pharmaceutically acceptable one-tenth salinization product thereof can be combined with medicinal common auxiliary material and carrier, the composition of preparation antitumor drug, thereby the effect that reaches prevention or treat.Said medicine can be selected suitable formulation according to the actual needs, as tablet, injection, suppository, aerosol, nanometer formulation etc.
The pharmaceutical composition that Toadpoison Medicine glycosylated derivative of the present invention and pharmaceutically acceptable one-tenth salinization product thereof are prepared into, can be used for treating kinds of tumors, comprising: prostate cancer, lung cancer, cancer of the stomach, liver cancer, mammary cancer, colorectal carcinoma, cervical cancer, skin carcinoma, nasopharyngeal carcinoma, oral carcinoma or leukemia.
The present invention has following advantage and effect with respect to prior art:
(1) the present invention can also increase water-soluble when keeping, having strengthened anti-tumor activity by Toadpoison Medicine being modified to its glycosylated derivative.And prove that through experiment in vitro Toadpoison Medicine glycosylated derivative provided by the present invention is compared with its parent compound Toadpoison Medicine, has reduced Normocellular toxic side effect.
(2) novel structure of Toadpoison Medicine glycosylated derivative of the present invention has good inhibition to tumour cell, good water solubility, and toxic side effect is low, possesses higher development and is worth.
(3) synthetic method security of the present invention is good, the productive rate height.
Embodiment
The present invention is described in further detail below in conjunction with embodiment, but embodiments of the present invention are not limited thereto.
Embodiment 1
Synthesizing of Toadpoison Medicine-3-N-D-glucoside, may further comprise the steps:
(1) gets bufalin(4.13g, 0.0107mol) place the 50ml round-bottomed flask, add CH
2Cl
2(35ml) be stirred to dissolving fully, slowly add PCC(pyridinium chlorochromate hydrochloride) (4.60g 0.0214mol), at the uniform velocity stirs 2.5h under the room temperature.Mixture is filtered, and filtrate decompression concentrates.Get reaction product Bufalinone(3.9g, 0.0102mol, productive rate 95% by silica gel column chromatography (the cyclohexane/ethyl acetate mixed solution of volume ratio 1:1)).
The product that obtains in the step (1) is carried out structural analysis.
Physico-chemical property: white powder (TLC R
f=0.35 cyclohexane/ethyl acetate 1:1).
Spectral information: ESI-MS m/z:385.5[M+H]
+, 407.4[M+Na]
+, 791.4[2M+H]
+, 823.3[2M+Na]
+ 1H?NMR（CDCl
3，300MHz）δ7.83（1H，dd，J=9.7，2.6Hz,H-22），7.22（1H，d，J=2.6Hz，H-21），6.23（1H，d，J=9.7Hz,H-23），2.62（1H，t，J=14.2Hz,H-4a,2.46（1H，dd，J=9.4,6.1Hz,H-17），2.33（m，1H），2.19（m，2H），2.19-1.20（m，19H），0.983（s，3H），0.703（s，3H）。
13C（CDCl
3，75MHz）δ212.9，162.5，148.6，146.9，122.7，115.3，85.0，51.2，48.4，43.7，42.15，42.13，40.7，37.1，36.8，36.7，35.2，32.7，28.7，26.6，22.6，21.5，21.0，16.6。
According to above spectral information, authenticating compound Bufalinone chemical formula is C
24H
32O
4, its structure is shown below.
(2) get Bufalinone(3.9g, 0.0102mol) place the 50ml round-bottomed flask, add 28mlCH
3OH is ultrasonic to add pyridine (4.88ml) to all dissolvings, and methoxamine hydrochloride (2.56g, 0.0306mol), stirring at room 2.5-3h.With mixture concentrating under reduced pressure solvent evaporated, with behind the HCl solution washing of 1mol/l, again with saturated common salt washing, CH
2Cl
2Extraction (4 * 15ml), MgSO
4Drying, the concentrating under reduced pressure organic layer gets oxime formula isomer mixture Bufalinone oximes(3.78g, 0.009mol, productive rate 90%).Get part oxime formula isomer mixture and separate, get the pure product IIa of oxime formula isomer and IIb.
The oxime formula isomer mixture that obtains in (2) is carried out structure to be identified.
The physico-chemical property of compound I Ia: white powder (TLC R
f=0.57 cyclohexane/ethyl acetate 1:1).
Spectral information: ESI-MS414.4[M+H]
+, 436.4[M+Na]
+, 849.3[2M+Na]
+ 1H?NMR（CDCl
3，300MHz）δ7.83（1H，dd，J=9.7，2.5Hz），7.21（1H，d，J=2.5Hz），6.23（1H，d，J=9.7Hz），3.77（s，3H），2.44（m，1H），2.00，1.68（m，2H），2.18，1.76（m，2H），2.12，1.84（m，2H），1.51，1.24（m，2H），1.68（m，H），1.50（m，H），0.91（s，3H），0.68（s，3H）。
13C?NMR（CDCl
3，75MHz）δ162.6，160.5，148.6，147.0，122.8，115.3，85.2，61.1，51.2，48.5，43.5，42.2，40.8，36.3，35.8，35.7，32.7，32.0，28.8，26.5，22.9，21.4，21.2，20.5，16.6。
According to above mass spectrum, one dimension spectral data and two-dimensional spectrum information, authenticating compound IIa chemical formula is C
25H
35NO
4, its structure is shown below.
The physico-chemical property of compound I Ib: white powder (TLC Rf=0.63 cyclohexane/ethyl acetate 1:1).
Spectral information: ESI-MS414.5[M+H]
+, 436.3[M+Na]
+, 849.5[2M+Na]
+ 1H?NMR（CDCl
3，300MHz）δ7.77（1H，dd，J=9.7，2.6Hz），7.15（1H，d，J=2.6Hz），6.17（1H，d，J=9.7Hz），3.73（s，3H），2.03，1.70（m，2H），2.19，1.74（m，2H），2.15，1.85（m，2H），1.89，1.23（m，2H），1.55，1.25（m，2H），2.46（m，H），1.65（m，H），1.54（m，H），0.87（s，3H），0.62（s，3H）。
13C?NMR（CDCl
3，75MHz）δ162.4，160.5，148.4，146.8，122.7，115.1，85.0，60.9，51.0，48.2，42.1，41.6，40.5，36.6，36.5，35.4，32.6，28.6，26.7，26.4，25.4，22.9，21.3，20.9，16.4。
According to above mass spectrum, one dimension spectral data and two-dimensional spectrum information, authenticating compound Bufalinone oxime IIb chemical formula is C
25H
35NO
4, its structure is shown below.
As seen, IIa and IIb are the configurational isomer of carbon-to-nitrogen double bon.
(3) get oxime formula mixture Bufalinone oximes(400mg, 0.969mmol) place 35ml reaction under high pressure pipe, add dioxane 3ml, dehydrated alcohol 1.5ml ultrasonicly dissolves fully to sample.After 20min to 0 ℃ of the ice bath cooling, place ice-water bath, add in the reaction tubes TERTIARY BUTYL AMINE base borane hydrochloride mixture (261.0mg 3.0mmol), slowly dropwise adds the 10%HCl aqueous solution (2.61ml) again, at the uniform velocity stir 3.5h after, add NaCO
3Termination reaction, CH
2Cl
2Extraction (4 * 15ml), MgSO
4Dry organic layer, concentrating under reduced pressure gets aglycon 3a, the 3b mixture.By silica gel column chromatography, the cyclohexane/ethyl acetate mixed solution gradient elution with volume ratio 4:1 obtains aglycon III(88.5mg, 0.213mmol, productive rate 22%), use eluent ethyl acetate at last, obtain aglycon IV(175mg, 0.426mmol, productive rate 43%).
The product III that obtains in (3) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.35 cyclohexane/ethyl acetate 1:1).
Spectral data: ESI-MS416.4[M+H]
+, 438.4[M+Na]
+, 831.4[2M+H]
+, 853.3[2M+Na]
+ 1H?NMR（CDCl
3，300MHz）δ7.84（1H，dd，J=9.7，2.5Hz），7.21（1H，d，J=2.5Hz），6.24（1H，d，J=9.7Hz），3.53（s，3H），3.24（s，1H），2.46（m，1H），2.17,1.70（m，2H），2.04，1.67（m，2H），1.44，1.33（m，2H），1.38，1.23（m，2H），1.45，1.35（m，2H），1.69，1.15（m，2H），1.86，1.27（m，2H），1.81，1.23（m，2H），1.84，1.35（m，2H），1.62（m，1H），1.51（m，1H），1.43（m，1H），0.90（s，3H），0.67（s，3H）。
13C?NMR（CDCl
3，75MHz）δ162.3，148.4，146.8，122.7，115.1，85.3，62.3，54.9，51.1，48.2，42.3，40.8，36.5，35.7，35.4，32.7，30.2，28.6，28.5，26.5，23.7，22.7，21.2，21.1，16.4。
According to above spectral information, authenticating compound III chemical formula is C
25H
37NO
4, its structure is shown below.
The product IV that obtains in (3) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.25 cyclohexane/ethyl acetate 1:1).
Wave spectrum character: ESI-MS416.3[M+H]
+, 438.3[M+Na]
+, 831.5[2M+H]
+, 853.4[2M+Na]
+ 1H?NMR（CDCl
3，300MHz）δ7.83（1H，dd，J=9.7，2.5Hz），7.21（1H，d，J=2.5Hz），6.24（1H，d，J=9.7Hz），3.54（s，3H），2.90（s，1H），2.43（m，1H），2.17,1.71（m，2H），2.06，1.66（m，2H），1.80，1.02（m，2H），1.83，1.36（m，2H），1.47，1.31（m，2H），1.71，1.27（m，2H），1.60，1.17（m，2H），1.54，1.45（m，2H），1.38，1.16（m，2H），1.63（m，1H），1.51（m，1H），1.40（m，1H），0.91（s，3H），0.67（s，3H）。
13C?NMR（CDCl
3，75MHz）δ162.4，148.4，146.8，122.7，115.2，85.3，62.6，60.3，51.1，48.2，42.5，41.4，40.8，36.4，35.4，35.1，32.7，31.0，28.6，27.1，25.3，23.4，21.5，21.1，16.4。
According to above spectral information, authenticating compound IV chemical formula is C
25H
37NO
4, its structure is shown below.
(4) get aglycon III(45mg, 0.108mmol), (39.0mg 0.217mmol) adds in the reaction tubes D-(+)-glucose, is dissolved in DMF/AcOH3:1 (1204 μ l), at the uniform velocity stirs 48h in 40 ℃.Reaction back mixture is removed reaction solvent through concentrating under reduced pressure, resistates is dissolved in the methyl alcohol, select preparative high performance liquid chromatography post (waters for use, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=15.02min obtains glycation product glycosides 1a(26mg, 0.045mmol, productive rate 42%).
The glycation product 1a that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.59 cyclohexane/ethyl acetate 3:7).
Wave spectrum character: ESI-MS m/z:578.5[M+H]
+, 600.5[M+Na]
+, 1177.5[2M+Na]
+ 1H?NMR（Pyridine-d
5，300MHz）δ8.23（1H，dd，J=9.7，2.5Hz），7.48（1H，d，J=2.5Hz），6.32（1H，d，J=9.7Hz），4.69（1H，d，J=8.7Hz），4.47（m，1H），4.26（m，1H），4.25（m，1H），3.89（m，1H），4.54，4.36（m，2H），3.96（s，3H），2.20，1.85（m，2H），2.02，1.86（m，2H），2.00，1.58（m，2H），1.43，1.33（m，2H），1.35，1.13（m，2H），1.63，1.07（m，2H），3.87（m，1H），2.49（m，1H），2.02（m，2H），1.87（m，2H），0.87（s，3H），0.82（s，3H）。
13C?NMR（Pyridine-d
5，75MHz）δ162.1，149.2，147.5，123.9，115.1，91.4，84.2，80.5，80.3，71.7，71.3，63.8，62.9，62.2，51.3，48.7，42.3，42.0，40.7，36.5，35.9，35.3，32.9，32.1，29.4，27.8，25.2，23.6，22.4，21.5，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound aglycon 1a chemical formula is C
31H
47NO
9, structure is shown below, and is Toadpoison Medicine-3 β-N-D-glucoside:
(5) get aglycon IV(50mg, 0.125mmol), D-(+)-Glucose(43.4mg 0.241mmol) adds in the reaction tubes, is dissolved in DMF/AcOH3:1 (1339 μ l), at the uniform velocity stirs 48h in 40 ℃.After the mixture concentrating under reduced pressure is removed reaction solvent after will reacting, resistates is dissolved in the methyl alcohol, by preparative high performance liquid chromatography post (waters, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=17.38min gets glycation product glycosides 1b(35mg, 0.061mmol, productive rate 48%).
The glycation product 1b that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.48 cyclohexane/ethyl acetate 3:7).
Spectral data: ESI-MS m/z578.4[M+H]
+, 600.3[M+Na]
+, 1177.3[2M+Na]
+ 1H?NMR（Pyridine-d
5，300MHz）δ8.33（1H，dd，J=9.7，2.5Hz），7.56（1H，d，J=2.5Hz），6.44（1H，d，J=9.7Hz），4.90（1H，d，J=8.7Hz），4.57（m，1H），4.39（m，1H），4.31（m，1H），3.98（m，1H），4.63，4.47（m，2H），4.06（s，3H），2.27，1.93（m，2H），1.90，1.60（m，2H），3.66（m，1H），2.58（m，1H），2.07（m，2H），2.26（m，2H），1.02（m，2H），0.97（s，3H），0.89（s，3H）。
13C?NMR（Pyridine-d
5，75MHz）δ162.1，149.2，147.5，123.9，115.1，91.4，84.2，80.5，80.3，71.7，71.3，63.8，62.9，62.2，51.3，48.7，42.3，42.0，40.7，36.5，35.9，35.3，32.9，32.1，29.4，27.8，25.2，23.6，22.4，21.5，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 1b chemical formula is C
31H
47NO
9Reaching structure and be shown below, is Toadpoison Medicine-3 α-N-D-glucoside.
The anti-tumor activity experiment of Toadpoison Medicine-3-N-D-glucoside comprises following project:
(1) adopt mtt assay detect Toadpoison Medicine-3-N-D-glucoside that present embodiment makes to prostate cancer cell DU145(available from American Type Culture Collection ATCC) the inhibition of proliferation effect.
Get the DU145 cell (RPMI1640 contains the 10%v/v foetal calf serum) of monolayer culture, with the trypsin solution digestion of mass volume ratio 0.25%, and be inoculated in (every sky adds cell suspending liquid 100 μ L, and cell count is 3000) on 96 orifice plates.Inoculate after 24 hours, add the Toadpoison Medicine-3-N-D-glucoside of different concns, and with equal volume than DMSO in contrast.Cultivate after 72 hours, every hole adds 20 μ lMTT solution (5mg/ml), the centrifugal supernatant liquor of abandoning after 4 hours, add DMSO(100 μ L/ hole), about vibration 15min, put microplate reader and measure the OD value, wavelength is 570nm, and the calculating cell survival rate, map simultaneously and try to achieve half-inhibition concentration (IC with SPASS13.0 software
50).
(2) adopt mtt assay detect Toadpoison Medicine-3-N-D-glucoside that present embodiment makes to prostate cancer cell PC3(available from American Type Culture Collection ATCC) the inhibition of proliferation effect.
Get the PC3 cell (RPMI1640 contains the 10%v/v foetal calf serum) of monolayer culture, with the trypsin solution digestion of mass volume ratio 0.25%, and be inoculated in (every sky adds cell suspending liquid 100 μ L, and cell count is 3000) on 96 orifice plates.Inoculate after 24 hours, add the Toadpoison Medicine-3-N-D-glucoside of different concns, and with equal volume than DMSO in contrast.Cultivate after 72 hours, every hole adds 20 μ L MTT solution (5mg/ml), the centrifugal supernatant liquor of abandoning after 4 hours, add DMSO(100 μ L/ hole), about vibration 15min, put microplate reader and measure the OD value, wavelength is 570nm, and the calculating cell survival rate, map simultaneously and try to achieve half-inhibition concentration (IC with SPASS13.0 software
50).
(3) adopt mtt assay detect Toadpoison Medicine-3-N-D-glucoside that present embodiment makes to hormonal dependent prostate cancer cell LNCaP(available from American Type Culture Collection ATCC) the inhibition of proliferation effect.
Get the LNCaP cell (DMEM contains the 10%v/v foetal calf serum) of monolayer culture, with the trypsin solution digestion of mass volume ratio 0.25%, and be inoculated in (every sky adds cell suspending liquid 100 μ L, and cell count is 3000) on 96 orifice plates.Inoculate after 24 hours, add the different concns testing sample, and with equal volume than DMSO in contrast.Cultivate after 72 hours, every hole adds 20 μ L MTT solution (5mg/ml), the centrifugal supernatant liquor of abandoning after 4 hours, add DMSO(100 μ L/ hole), about vibration 15min, put microplate reader and measure the OD value, wavelength is 570nm, and the calculating cell survival rate, map simultaneously and try to achieve half-inhibition concentration (IC with SPASS13.0 software
50).
(4) adopt mtt assay detect Toadpoison Medicine-3-N-D-glucoside that present embodiment makes to breast cancer cell MCF-7(available from American Type Culture Collection ATCC) the inhibition of proliferation effect.
Get the MCF-7 cell (RPMI1640 contains the 10%v/v foetal calf serum) of monolayer culture, with the trypsin solution digestion of mass volume ratio 0.25%, and be inoculated in (every sky adds cell suspending liquid 100 μ L, and cell count is 3000) on 96 orifice plates.Inoculate after 24 hours, add the Toadpoison Medicine-3-N-D-glucoside of different concns, and with equal volume than DMSO in contrast.Cultivate after 72 hours, every hole adds 20 μ LMTT solution (5mg/ml), the centrifugal supernatant liquor of abandoning after 4 hours, add DMSO(100 μ L/ hole), about vibration 15min, put microplate reader and measure the OD value, wavelength is 570nm, and the calculating cell survival rate, map simultaneously and try to achieve half-inhibition concentration (IC with SPASS13.0 software
50).
(5) adopt mtt assay detect Toadpoison Medicine-3-N-D-glucoside that present embodiment makes to adenocarcinoma of the uterine cervix cell HeLa(available from American Type Culture Collection ATCC) the inhibition of proliferation effect.
Get the HeLa cell (RPMI1640 contains the 10%v/v foetal calf serum) of monolayer culture, with the trypsin solution digestion of mass volume ratio 0.25%, and be inoculated in (every sky adds cell suspending liquid 100 μ L, and cell count is 3000) on 96 orifice plates.Inoculate after 24 hours, add the Toadpoison Medicine-3-N-D-glucoside of different concns, and with equal volume than DMSO in contrast.Cultivate after 72 hours, every hole adds 20 μ L MTT solution (5mg/ml), the centrifugal supernatant liquor of abandoning after 4 hours, add DMSO(100 μ L/ hole), about vibration 15min, put microplate reader and measure the OD value, wavelength is 570nm, and the calculating cell survival rate, map simultaneously and try to achieve half-inhibition concentration (IC with SPASS13.0 software
50).
(6) adopt mtt assay detect Toadpoison Medicine-3-N-D-glucoside that present embodiment makes to normal cell Vero(available from American Type Culture Collection ATCC) the inhibition of proliferation effect.
Get the Vero cell (RPMI1640 contains the 10%v/v foetal calf serum) of monolayer culture, with the trypsin solution digestion of mass volume ratio 0.25%, and be inoculated in (every sky adds cell suspending liquid 100 μ L, and cell count is 3000) on 96 orifice plates.Inoculate after 24 hours, add the Toadpoison Medicine-3-N-D-glucoside of different concns, and with equal volume than DMSO in contrast.Cultivate after 72 hours, every hole adds 20 μ L MTT solution (5mg/ml), the centrifugal supernatant liquor of abandoning after 4 hours, add DMSO(100 μ L/ hole), about vibration 15min, put microplate reader and measure the OD value, wavelength is 570nm, and the calculating cell survival rate, map simultaneously and try to achieve half-inhibition concentration (IC with SPASS13.0 software
50).
Adopt aforesaid method to compare Toadpoison Medicine-3 β-N-D-glucoside (1a) and Toadpoison Medicine-3 α-N-D-glucoside (1b) to tumour cell DU145, PC3, LNCaP, MCF-7, the inhibition activity of HeLa and normal cell Vero.Wherein glycation product 1a is to the half-inhibition concentration (IC of above-mentioned tumor cell line
50) be respectively 0.48 ± 0.13,0.32 ± 0.12,0.51 ± 0.09,0.71 ± 0.12,0.90 ± 0.14 μ M, and to the IC of normal cell Vero
50Value is greater than 20 μ M.Glycation product 1b is to the IC of above-mentioned tumor cell line
50Value is respectively 1.82 ± 0.21,2.44 ± 0.31,2.05 ± 0.20,2.01 ± 0.12,2.32 ± 0.23 μ M.Similar to 1a, the IC of the normal cell Vero of 1b
50Value is also greater than 20 μ M.Toadpoison Medicine is to the IC of above-mentioned tumor cell line
50Value is 0.34 ± 0.06,0.58 ± 0.04,0.67 ± 0.05,0.49 ± 0.08,0.54 ± 0.07 μ M, and Toadpoison Medicine is to the IC of normal cell Vero
50Value is 0.71 ± 0.14 μ M.
Toadpoison Medicine-3-N-D-glucoside profit partition ratio determination experiment
Adopt classical fask oscillating method to measure the profit partition ratio (logP) of Toadpoison Medicine-3 β-N-D-glucoside (1a) and Toadpoison Medicine-3 α-N-D-glucoside (1b), the logP of 1a is 1.915, the logP of 1b is 1.926, and the logP of Toadpoison Medicine is 3.625 under the same terms.As seen, 1a and 1b's is water-soluble all good than Toadpoison Medicine.
Brief summary: as can be seen from the above results, the anti-tumor activity of Toadpoison Medicine-3 β-N-D-glucoside (1a) is similar to Toadpoison Medicine.And the anti-tumor activity of Toadpoison Medicine-3 α-N-D-glucoside (1b) is weaker than Toadpoison Medicine, but two kinds of D-glucosides are all low than Toadpoison Medicine to Normocellular toxicity.And 1a and 1b's is water-soluble all good than Toadpoison Medicine.
Embodiment 2
Synthesizing of Toadpoison Medicine-L-glucoside, may further comprise the steps:
(1)-(3) with preparation aglycon method among the embodiment 1.
(4) get aglycon III(46mg, 0.108mmol), (39.0mg 0.217mmol) adds in the reaction tubes L-glucose, is dissolved in DMF/AcOH3:1 (1204 μ l), at the uniform velocity stirs 48h in 40 ℃.Reaction back mixture concentrating under reduced pressure drying is removed reaction solvent, sample is dissolved in the methyl alcohol, select preparative high performance liquid chromatography post (waters for use, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=16.02min obtains glycation product glycosides 2a(26mg, 0.045mmol, productive rate 42%).
The glycation product 2a that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.61 cyclohexane/ethyl acetate 3:7).
Wave spectrum character: ESI-MS m/z:578.4[M+H]
+, 600.5[M+Na]
+, 1177.2[2M+Na]
+ 1H?NMR（Pyridine-d
5，400MHz）δ8.25（1H，dd，J=9.6，2.4Hz），7.48（1H，d，J=2.4Hz），6.35（1H，d，J=9.6Hz），4.72（1H，d，J=8.6Hz），4.46（m，1H），4.28（m，1H），4.22（m，1H），3.88（m，1H），4.55，4.35（m，2H），3.94（s，3H），3.83（m，1H），2.50（m，1H），2.18，1.87（m，2H），2.16，1.45（m，2H），1.79，1.33（m，2H），1.69，1.51（m，2H），1.28，1.09（m，2H），2.19（m，1H），1.87（m，1H），1.73（m，1H），1.75（m，1H），1.42（m，1H），0.89（s，3H），0.81（s，3H）。
13C?NMR（Pyridine-d
5，100MHz）δ162.1，149.2，147.5，123.8，115.1，90.9，84.3，80.5，80.2，71.7，71.6，63.6，62.9，57.1，51.4，48.8，42.2，40.8，37.0，36.5，35.8，32.8，31.0，29.5，29.4，27.2，23.9，23.7，22.1，21.6，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 2a is Toadpoison Medicine-3 α-N-L-glucoside, and its chemical formula is C
31H
47NO
9, structure is shown below.
(5) get aglycon IV(50mg, 0.125mmol), L-(-)-Glucose(43.4mg 0.241mmol) adds in the reaction tubes, is dissolved in DMF/AcOH3:1 (1.3ml), at the uniform velocity stirs 48h in 40 ℃.After mixture concentrating under reduced pressure drying is removed reaction solvent after will reacting, sample is dissolved in the methyl alcohol, by preparative high performance liquid chromatography post (waters, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=13.29min gets glycation product glycosides 2b(35mg, 0.061mmol, productive rate 48%).
The glycation product 2b that obtains in (5) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.44 cyclohexane/ethyl acetate 3:7).
Spectral data: ESI-MS m/z578.3[M+H]
+, 600.3[M+Na]
+, 1177.0[2M+Na]
+ 1H?NMR（Pyridine-d
5，400MHz）δ8.24（1H，dd，J=9.7，2.5Hz），7.47（1H，d，J=2.5Hz），6.35（1H，d，J=9.7Hz），4.88（1H，d，J=8.8Hz），4.53（m，1H），4.38（m，1H），4.28（m，1H），3.61（m，1H），4.49，4.25（m，2H），3.98（s，3H），2.16，1.85（m，2H），2.15-0.90（m，19H），0.89（s，3H），0.81（s，3H）。
13C?NMR（Pyridine-d
5，100MHz）δ162.1，149.2，147.5，123.9，115.1，91.1，84.2，80.5，80.3，71.7，71.4，63.8，62.9，62.0，51.4，48.7，42.3，42.1，40.8，36.5，35.6，35.2，32.8，30.9，29.4，27.8，26.4，23.6，22.2，21.5，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 2b is Toadpoison Medicine-3 α-N-L-glucoside, and its chemical formula is C
31H
47NO
9, structure is shown below.
The anti-tumor activity experiment of Toadpoison Medicine-L-glucoside may further comprise the steps:
Method same among employing and the embodiment 1 detects glycation product 2a and the tumour cell DU145 of 2b, PC3, LNCaP, MCF-7, the inhibition activity of HeLa and normal cell Vero.Wherein glycation product glycation product 2a is to the IC of above-mentioned tumor cell line
50Value is respectively 0.16 ± 0.05,0.49 ± 0.12,0.42 ± 0.09,0.59 ± 0.14,0.88 ± 0.14 μ M.And to the IC of the normal cell Vero of 2a
50Value is 11.7 ± 1.3 μ M.2b is to the half-inhibition concentration (IC of above-mentioned tumor cell line
50) be respectively 1.47 ± 0.11,2.46 ± 0.103,1.15 ± 0.40,1.66 ± 0.32,2.43 ± 0.13 μ M.And the IC of the normal cell Vero of 2b
50Value is greater than 20 μ M.Toadpoison Medicine is to the IC of above-mentioned tumor cell line
50Value is 0.34 ± 0.06,0.58 ± 0.04,0.67 ± 0.05,0.49 ± 0.08,0.54 ± 0.07 μ M.Toadpoison Medicine is to the IC of normal cell Vero
50Value is 0.71 ± 0.14 μ M.
Toadpoison Medicine-L-glucoside profit partition ratio is measured
Adopt classical fask oscillating method to measure the profit partition ratio (logP) of Toadpoison Medicine-3 β-N-L-glucoside (2a) and Toadpoison Medicine-3 α-N-L-glucoside (2b), the logP of 2a is 1.896, the logP of 2b is 1.913, and the logP of Toadpoison Medicine is 3.625 under the same terms.As seen, 2a and 2b's is water-soluble all good than Toadpoison Medicine.
Brief summary: by above result as can be seen, the anti-tumor activity of Toadpoison Medicine-3 β-N-L-glucoside (2a) is similar to Toadpoison Medicine, and the anti-tumor activity of Toadpoison Medicine-3 α-N-L-glucoside (2b) is weaker than Toadpoison Medicine.But two kinds of L-glucosides are all low than Toadpoison Medicine to Normocellular toxicity.And 2a and 2b's is water-soluble all good than Toadpoison Medicine.
Embodiment 3
Synthesizing of Toadpoison Medicine-D-fucoside, may further comprise the steps:
(1)-(3) with the step among the embodiment 1.
(4) get aglycon III(42mg, 0.101mmol), (33.6mg 0.205mmol) adds in the reaction tubes D-Fucose, is dissolved in DMF/AcOH3:1 (1122 μ l), at the uniform velocity stirs 48h in 40 ℃.Reaction back mixture concentrating under reduced pressure drying is removed reaction solvent, sample is dissolved in the methyl alcohol, select preparative high performance liquid chromatography post (waters for use, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=11.57min obtains glycation product glycosides 3a(22mg, 0.039mmol, productive rate 39%).
The glycation product 3a that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.52 cyclohexane/ethyl acetate 3:7)
Wave spectrum character: ESI-MS m/z:562.4[M+H]
+, 584.4[M+Na]
+, 1145.3[2M+Na]
+ 1H（Pyridine-d
5，300MHz）δ8.25（1H，dd，J=9.7，2.5Hz），7.48（1H，d，J=2.5Hz），6.35（1H，d，J=9.7Hz），4.51（1H，d，J=8.7Hz），4.69（m，1H），4.15（m，1H），4.10（m，1H），3.83（m，1H），3.88（s，3H），2.19，1.89（m，2H），2.14，1.56（m，2H），2.13，1.75（m，2H），1.67，1.26（m，2H），1.43，1.13（m，2H），3.80（m，1H），2.51（m，1H），1.83（m，1H），2.28（m，2H），1.91（m，2H），1.78（m，1H），1.45（m，2H），1.59（d，3H，J=6.4Hz），0.89（s，3H），0.84（s，3H）。
13C（Pyridine-d
5，75MHz）δ162.1，149.2，147.5，123.9，115.1，91.4，84.3，77.1，73.1，72.8，68.3，63.3，57.2，51.4，48.8，42.3，40.8，37.1，36.3，35.9，32.8，31.1，30.0，29.4，27.6，24.0，23.9，21.8，21.7，17.4，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound aglycon 3a chemical formula is C
31H
47NO
8, structure is shown below.
(5) get aglycon IV(51mg, 0.125mmol), D-(+)-Fucose(38.2mg 0.233mmol) adds in the reaction tubes, is dissolved in DMF/AcOH3:1 (1300 μ l), at the uniform velocity stirs 48h in 40 ℃.After mixture concentrating under reduced pressure drying is removed reaction solvent after will reacting, sample is dissolved in the methyl alcohol, by preparative high performance liquid chromatography post (waters, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=13.81min gets glycation product glycosides 3b(35mg, 0.062mmol, productive rate 49%).
The glycation product 3b that obtains in (5) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.42 cyclohexane/ethyl acetate 3:7).
Spectral data: ESI-MS m/z562.3[M+H]
+, 584.3[M+Na]
+, 1145.3[2M+Na]
+ 1H?NMR（Pyridine-d
5，400MHz）δ8.23（1H，dd，J=9.7，2.5Hz），7.47（1H，d，J=2.5Hz），6.36（1H，d，J=9.7Hz），4.62（1H，d，J=8.7Hz），4.73（m，1H），4.15（m，1H），4.10（m，1H），3.87（s，3H），3.68（m，1H），2.51（m，1H），2.20，1.89（m，2H），2.20-0.90（m，19H）,1.58（d，3H，J=6.3Hz），0.90（s，3H），0.86（s，3H）。
13C?NMR（Pyridine-d
5，100MHz）δ162.1，149.2，147.5，123.9，115.2，91.9，84.2，77.2，73.1，72.8，68.5，63.5，62.2，51.3，48.7，42.4，40.8，36.58，36.52，35.2，32.9，32.2，31.4，29.4，27.7，25.3，23.6，22.3，21.5，17.5，17.2。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 3b is Toadpoison Medicine-3 α-N-D-fucoside, and its chemical formula is C
31H
47NO
8, structure is shown below.
The anti-tumor activity experiment of Toadpoison Medicine-D-fucoside
Method same among employing and the embodiment 1 detects glycation product 3a and the tumour cell DU145 of 3b, PC3, LNCaP, MCF-7, the inhibition activity of HeLa and normal cell Vero.Wherein glycation product 3a is to the IC of above-mentioned tumor cell line
50Value is respectively 0.55 ± 0.02,0.25 ± 0.05,0.36 ± 0.09,0.39 ± 0.10,0.85 ± 0.12 μ M.And to the IC of the normal cell Vero of 3a
50Value is greater than 20 μ M.Glycation product 3b is to the half-inhibition concentration (IC of above-mentioned tumor cell line
50) be respectively 2.01 ± 0.13,4.68 ± 0.24,1.58 ± 0.21,2.06 ± 0.10,3.32 ± 0.34 μ M.And the IC of the normal cell Vero of 3b
50Value is greater than 20 μ M.Toadpoison Medicine is to the IC of above-mentioned tumor cell line
50Value is 0.34 ± 0.06,0.58 ± 0.04,0.67 ± 0.05,0.49 ± 0.08,0.54 ± 0.07 μ M.Toadpoison Medicine is to the IC of normal cell Vero
50Value is 0.71 ± 0.14 μ M.
The profit partition ratio of Toadpoison Medicine-D-fucoside is measured
Adopt classical fask oscillating method to measure the profit partition ratio (logP) of Toadpoison Medicine-3 β-N-D-fucoside (3a) and Toadpoison Medicine-3 α-N-D-fucoside (3b), the logP of 3a is 2.902, the logP of 3b is 2.918, and the logP of Toadpoison Medicine is 3.625 under the same terms.As seen, 3a and 3b's is water-soluble all good than Toadpoison Medicine.
Brief summary: by above result as can be seen, anti-tumor activity and the Toadpoison Medicine of Toadpoison Medicine-3 β-N-D-fucoside (3a) are suitable, and the anti-tumor activity of Toadpoison Medicine-3 α-N-D-fucoside (3b) is weaker than Toadpoison Medicine.But two kinds of D-fucosides are all low than Toadpoison Medicine to Normocellular toxicity.And 3a and 3b's is water-soluble all good than Toadpoison Medicine.
Embodiment 4
Synthesizing of Toadpoison Medicine-L-fucoside, may further comprise the steps:
(1)-(3) with preparation aglycon method among the embodiment 1.
(4) get aglycon III(45mg, 0.108mmol), (35.6mg 0.217mmol) adds in the reaction tubes L-Fucose, is dissolved in DMF/AcOH3:1 (1204 μ l), at the uniform velocity stirs 48h in 40 ℃.Reaction back mixture concentrating under reduced pressure drying is removed reaction solvent, sample is dissolved in the methyl alcohol, select preparative high performance liquid chromatography post (waters for use, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile solution with 45% is as moving phase, retention time t
R=16.52min obtains glycation product glycosides 4a(27mg, 0.048mmol, productive rate 45%).
The glycation product 4a that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.60 cyclohexane/ethyl acetate 3:7).
Spectral data: ESI-MS m/z:562.5[M+H]
+, 584.3[M+Na]
+, 1145.2[2M+Na]
+ 1H?NMR（Pyridine-d
5，400MHz）δ8.23（1H，dd，J=9.7，2.4Hz），7.47（1H，d，J=2.4Hz），6.34（1H，d，J=9.7Hz），4.49（1H，d，J=8.7Hz），4.66（m，1H），4.11（m，1H），4.07（m，1H），3.72（m，1H），3.81（s，3H），2.17，1.86（m，2H），2.11，1.88（m，2H），1.81，1.32（m，2H），1.45，1.38（m，2H），1.37，1.14（m，2H），3.79（m，1H），2.50（m，1H），1.78（m，1H），2.16（m，2H），2.15（m，2H），1.98（m，2H），1.78（m，1H），1.65（m，2H），1.53（s，3H，J=6.3Hz），0.90（s，3H），0.84（m，3H）。
13C?NMR（Pyridine-d
5，100MHz）δ162.1，149.2，147.5，123.9，115.2，91.3，84.4，77.1，73.0，72.8，68.7，63.3，57.1，51.4，48.8，42.3，40.9，37.1，36.6，35.9，32.9，31.2，29.6，29.4，27.2，24.0，23.8，22.1，21.7，17.3，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 4a is Toadpoison Medicine-3 β-N-L-fucoside, and its chemical formula is C
31H
47NO
8, structure is shown below.
(5) get aglycon IV(45mg, 0.108mmol), (38mg 0.232mmol) adds in the reaction tubes L-Fucose, is dissolved in DMF/AcOH3:1 (1205 μ l), at the uniform velocity stirs 48h in 40 ℃.After mixture concentrating under reduced pressure drying is removed reaction solvent after will reacting, sample is dissolved in the methyl alcohol, by preparative high performance liquid chromatography post (waters, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=14.38min gets glycation product glycosides 4b(31mg, 0.055mmol, productive rate 51%).
The glycation product 4b that obtains in (5) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.47 cyclohexane/ethyl acetate 3:7).
Spectral data: ESI-MS m/z562.2[M+H]
+, 584.2[M+Na]
+, 1145.1[2M+Na]
+ 1H?NMR（Pyridine-d
5，400MHz）δ8.23（1H，dd，J=9.7，2.5Hz），7.47（1H，d，J=2.5Hz），6.36（1H，d，J=9.7Hz），4.63（1H，d，J=8.7Hz），4.71（m，1H），4.20（m，1H），4.10（m，1H），3.96（s，3H），3.67（m，1H），2.49（m，1H），2.19，1.87（m，2H），2.18-0.90（m，19H）,1.57（d，3H，J=6.4Hz），0.88（s，3H），0.81（s，3H）。
13C?NMR（Pyridine-d
5，100MHz）δ162.1，149.2，147.5，123.9，115.2，91.9，84.2，77.2，73.1，72.8，68.5，63.5，62.2，51.3，48.7，42.4，40.8，36.58，36.52，35.2，32.9，32.2，31.4，29.4，27.7，25.3，23.6，22.3，21.5，17.5，17.2。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 4b is Toadpoison Medicine-3 α-N-L-fucoside, and its chemical formula is C
31H
47NO
8, structure is shown below.
The anti-tumor activity experiment of Toadpoison Medicine-L-Fucose glucosides
Method same among employing and the embodiment 1 detects glycation product 4a and the tumour cell DU145 of 4b, PC3, LNCaP, MCF-7, the inhibition activity of HeLa and normal cell Vero.Wherein glycation product 4a is to the IC of above-mentioned tumor cell line
50Value is respectively 0.22 ± 0.01,0.12 ± 0.04,0.32 ± 0.06,0.55 ± 0.14,0.45 ± 0.10 μ M, and to the IC of the normal cell Vero of 4a
50Value is 10.8 ± 1.2 μ M.Glycation product 4b is to the half-inhibition concentration (IC of above-mentioned tumor cell line
50) be respectively 1.46 ± 0.04,2.07 ± 0.53,1.25 ± 0.12,1.69 ± 0.22,2.42 ± 0.20 μ M, the IC of the normal cell Vero of 4b
50Value is greater than 20 μ M.Toadpoison Medicine is to the IC of above-mentioned tumor cell line
50Value is 0.34 ± 0.06,0.58 ± 0.04,0.67 ± 0.05,0.49 ± 0.08,0.54 ± 0.07 μ M.Toadpoison Medicine is to the IC of normal cell Vero
50Value is 0.71 ± 0.14 μ M.
Toadpoison Medicine-L-Fucose glucosides profit partition ratio is measured
Adopt classical fask oscillating method to measure the profit partition ratio (logP) of Toadpoison Medicine-3 β-N-L-fucoside (4a) and Toadpoison Medicine-3 α-N-L-fucoside (4b), the logP of 4a is 2.898, the logP of 4b is 2.907, and the logP of Toadpoison Medicine is 3.625 under the same terms.As seen, 4a and 4b's is water-soluble all good than Toadpoison Medicine.
Brief summary: by above result as can be seen, the anti-tumor activity of Toadpoison Medicine-3 β-N-L-fucoside (4a) is similar to Toadpoison Medicine, and the anti-tumor activity of Toadpoison Medicine-3 α-N-D-fucoside (4b) is weaker than Toadpoison Medicine.But two kinds of L-fucosides are all low than Toadpoison Medicine to Normocellular toxicity.And 4a and 4b's is water-soluble all good than Toadpoison Medicine.
Embodiment 5
Synthesizing of Toadpoison Medicine-D-lyxoside, may further comprise the steps:
(1)-(3) with preparation aglycon method among the embodiment 1.
(4) get aglycon III(41mg, 0.099mmol), (28.9mg 0.217mmol) adds in the reaction tubes D-lyxose, is dissolved in DMF/AcOH3:1 (1098 μ l), at the uniform velocity stirs 48h in 40 ℃.Reaction back mixture concentrating under reduced pressure drying is removed reaction solvent, sample is dissolved in the methyl alcohol, select preparative high performance liquid chromatography post (waters for use, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=11.65min obtains glycation product glycosides 5a(29mg, 0.053mmol, productive rate 54%).
The glycation product 5a that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.63 cyclohexane/ethyl acetate 3:7)
Wave spectrum character: ESI-MS m/z:548.1[M+H]
+, 570.3[M+Na]
+, 1117.2[2M+Na]
+ 1H（Pyridine-d
5，400MHz）δ8.25（1H，dd，J=9.9，2.8Hz），7.48（1H，d，J=2.8Hz），6.35（1H，d，J=9.9Hz），4.71（1H），4.81（m，1H），4.38（m，1H），4.21（m，1H），4.40，3.64（m，2H），3.89（m，1H），3.92（s，3H），2.46（m，1H），2.16，1.83（m，2H），2.08，1.34（m，2H），1.80，1.35（m，2H），1.29，1.08（m，2H），2.03（m，2H），1.77（m，1H），1.81（m，1H），1.49（m，2H），1.35（m，2H），0.89（s，3H），0.79（s，3H）。
13C（Pyridine-d
5，100MHz）δ162.1，149.2，147.5，123.0，115.1，84.3，82.6，73.2，71.3，67.8，63.3，61.8，57.2，51.4，48.8，42.3，40.8，36.59，36.53，35.8，32.8，31.0，29.7，29.4，27.,2，23.8，22.1，21.6，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 5a is Toadpoison Medicine-3 β-N-D-lyxoside, and its chemical formula is C
30H
45NO
8, structure is shown below.
The anti-tumor activity experiment of Toadpoison Medicine-D-lyxoside
Method same among employing and the embodiment 1 detects the tumour cell DU145 of glycation product 5a, PC3, LNCaP, MCF-7, the inhibition activity of HeLa and normal cell Vero.Wherein glycation product 5a is to the half-inhibition concentration (IC of above-mentioned tumor cell line
50) be respectively 0.75 ± 0.10,0.48 ± 0.03,0.36 ± 0.08,0.28 ± 0.08,0.48 ± 0.09 μ M, and the IC of the normal cell Vero of 5a
50Value is 10.40 ± 1.20 μ M.Toadpoison Medicine is to the IC of above-mentioned tumor cell line
50Value is 0.34 ± 0.06,0.58 ± 0.04,0.67 ± 0.05,0.49 ± 0.08,0.54 ± 0.07 μ M.Toadpoison Medicine is to the IC of normal cell Vero
50Value is 0.71 ± 0.14 μ M.
Toadpoison Medicine-D-lyxoside profit partition ratio is measured
Adopt classical fask oscillating method to measure the profit partition ratio (logP) of Toadpoison Medicine-3 β-N-D-lyxoside (5a), the logP of 5a is 2.313, and the logP of Toadpoison Medicine is 3.625 under the same terms.As seen, 5a's is water-soluble good than Toadpoison Medicine.
Brief summary: by above result as can be seen, anti-tumor activity and the Toadpoison Medicine of Toadpoison Medicine-3 β-N-D-lyxoside (5a) are suitable, but low than Toadpoison Medicine to Normocellular toxicity.And 5a's is water-soluble all good than Toadpoison Medicine.
Embodiment 6
Synthesizing of Toadpoison Medicine-D-Arabinoside, may further comprise the steps:
(1)-(3) with preparation aglycon method among the embodiment 1.
(4) get aglycon 3b(40mg, 0.096mmol), (28.9mg 0.192mmol) adds in the reaction tubes D-pectinose, is dissolved in DMF/AcOH3:1 (1071 μ l), at the uniform velocity stirs 48h in 40 ℃.Reaction back mixture concentrating under reduced pressure drying is removed reaction solvent, sample is dissolved in the methyl alcohol, select preparative high performance liquid chromatography post (waters for use, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile solution with 45% is as moving phase, retention time t
R=15.52min obtains glycation product glycosides 6a(28mg, 0.051mmol, productive rate 53%).
The glycation product 6a that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.57 cyclohexane/ethyl acetate 3:7).
Spectral data: ESI-MS m/z:548.5[M+H]
+, 570.5[M+Na]
+, 1117.6[2M+Na]
+ 1H?NMR（Pyridine-d
5，400MHz）δ8.25（1H，dd，J=9.7，2.4Hz），7.46（1H，d，J=2.4Hz），6.36（1H，d，J=9.7Hz），4.50（1H，d，J=8.7Hz），4.77（m，1H），4.29（m，2H），4.16（m，1H），3.71（m，1H），3.86（s，3H），2.17，1.84（m，2H），1.81，1.27（m，2H），1.42，1.14（m，2H），1.35，1.12（m，2H），3.79（m，1H），2.48（m，1H），1.79（m，1H），1.93（m，2H），1.84（m，1H），1.46（m，2H），0.90（s，3H），0.84（m，3H）。
13C?NMR（Pyridine-d
5，100MHz）δ162.1，149.2，147.5，123.9，115.2，91.8，84.3，76.5，70.2，69.3，69.2，63.3，57.3，51.4，48.8，42.3，40.8，37.1，36.6，35.9，32.8，31.2，29.7，29.4，27.2，24.1，23.8，22.1，21.7，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 6a is Toadpoison Medicine-3 β-N-D-Arabinoside, and its chemical formula is C
30H
45NO
8, structure is shown below.
The anti-tumor activity experiment of Toadpoison Medicine-D-Arabinoside
Method same among employing and the embodiment 1 detects the tumour cell DU145 of glycation product 6a, PC3, LNCaP, MCF-7, the inhibition activity of HeLa and normal cell Vero.Wherein glycation product 6a is to the half-inhibition concentration (IC of above-mentioned tumor cell line
50) be respectively 0.80 ± 0.10,0.36 ± 0.07,0.17 ± 0.05,0.19 ± 0.04,0.32 ± 0.05 μ M, and the IC of the normal cell Vero of 6a
50Value is 11.3 ± 0.80 μ M.Toadpoison Medicine is to the IC of above-mentioned tumor cell line
50Value is 0.34 ± 0.06,0.58 ± 0.04,0.67 ± 0.05,0.49 ± 0.08,0.54 ± 0.07 μ M.Toadpoison Medicine is to the IC of normal cell Vero
50Value is 0.71 ± 0.14 μ M.
Toadpoison Medicine-D-Arabinoside profit partition ratio is measured
Adopt classical fask oscillating method to measure the profit partition ratio (logP) of Toadpoison Medicine-3 β-N-D-Arabinoside (6a), the logP of 6a is 2.324, and the logP of Toadpoison Medicine is 3.625 under the same terms.As seen, 6a's is water-soluble good than Toadpoison Medicine.
Brief summary: by above result as can be seen, anti-tumor activity and the Toadpoison Medicine of Toadpoison Medicine-3 β-N-D-Arabinoside (6a) are suitable, but low than Toadpoison Medicine to Normocellular toxicity.And 6a's is water-soluble all good than Toadpoison Medicine.
Embodiment 7
Synthesizing of Toadpoison Medicine-D-xyloside, may further comprise the steps:
(1)-(3) with preparation aglycon method among the embodiment 1.
(4) get aglycon III(43mg, 0.104mmol), (28.9mg 0.192mmol) adds in the reaction tubes D wood sugar, is dissolved in DMF/AcOH3:1 (1151 μ l), at the uniform velocity stirs 48h in 40 ℃.Reaction back mixture concentrating under reduced pressure drying is removed reaction solvent, sample is dissolved in the methyl alcohol, select preparative high performance liquid chromatography post (waters for use, C18,5 μ m, 20 * 250mm) separation and purification samples, flow velocity 8ml/min, detect wavelength 296nm, the acetonitrile/water solution with 45% is as moving phase, retention time t
R=12.49min obtains glycation product glycosides 7a(22mg, 0.040mmol, productive rate 38%).
The glycation product 7a that obtains in (4) is carried out structure to be identified.
Physico-chemical property: white powder (TLC R
f=0.54 cyclohexane/ethyl acetate 3:7)
Wave spectrum character: ESI-MS m/z:548.5[M+H]
+, 570.6[M+Na]
+, 1117.7[2M+Na]
+ 1H（Pyridine-d
5，400MHz）δ8.23（1H，dd，J=9.7，2.5Hz），7.47（1H，d，J=2.5Hz），6.35（1H，d，J=9.7Hz），4.59（1H，d，J=8.6Hz），4.43（m，1H），4.25（m，1H），4.18（m，1H），4.41，3.69（m，2H），3.98（s，3H），3.85（m，1H），2.10，1.86（m，2H），2.0，1.39（m，2H），1.97，1.50（m，2H），2.50（m，1H），2.16（m，H），1.86（m，1H），1.78（m，1H），1.75（m，1H），1.69（m，1H），1.42（m，1H），0.89（s，3H），0.84（d，3H）。
13C（Pyridine-d
5，100MHz）δ162.0，149.3，147.5，123.8，115.1，92.0，84.3，80.5，71.5，71.0，69.1，63.6，57.4，51.5，48.8，42.3，40.9，37.1，35.9，35.8，32.9，31.1，30.1，29.4，27.5，24.1，24.0，23.2，21.8，17.1。
According to mass spectrum, nucleus magnetic resonance one dimension and two-dimensional spectrum information, authenticating compound 7a is Toadpoison Medicine-3 β-N-D-xyloside, and its chemical formula is C
30H
45NO
8, structure is shown below.
The anti-tumor activity experiment of Toadpoison Medicine-D-xyloside
Method same among employing and the embodiment 1 detects the tumour cell DU145 of glycation product 7a, PC3, LNCaP, MCF-7, the inhibition activity of HeLa and normal cell Vero.Glycation product 7a is to the IC of above-mentioned tumor cell line
50Value is respectively 0.24 ± 0.06,0.56 ± 0.03,0.35 ± 0.08,0.28 ± 0.16,0.86 ± 0.13 μ M.And to the IC of the normal cell Vero of 7a
50Value is 14.7 ± 2.6 μ M.Toadpoison Medicine is to the IC of above-mentioned tumor cell line
50Value is 0.34 ± 0.06,0.58 ± 0.04,0.67 ± 0.05,0.49 ± 0.08,0.54 ± 0.07 μ M.Toadpoison Medicine is to the IC of normal cell Vero
50Value is 0.71 ± 0.14 μ M.
Toadpoison Medicine-D-xyloside profit partition ratio is measured
Adopt classical fask oscillating method to measure the profit partition ratio (logP) of Toadpoison Medicine-3 β-N-D-xyloside (7a), the logP of 7a is 2.338, and the logP of Toadpoison Medicine is 3.625 under the same terms.As seen, 7a's is water-soluble good than Toadpoison Medicine.
Brief summary: by above result as can be seen, anti-tumor activity and the Toadpoison Medicine of Toadpoison Medicine-3 β-N-D-xyloside (7a) are suitable, but low than Toadpoison Medicine to Normocellular toxicity.And 7a's is water-soluble all good than Toadpoison Medicine.
Above-described embodiment is preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. Toadpoison Medicine glycosylated derivative, be characterised in that: its structure is suc as formula shown in I or the formula II:
Wherein R is the reducing sugar glycosyl.
2. Toadpoison Medicine glycosylated derivative according to claim 1, it is characterized in that: R is D-glucose sugar, L-glucose sugar, Fucose, wood sugar, lyxose or Arabic glycosyl.
3. the synthetic method of claim 1 or 2 described Toadpoison Medicine glycosylated derivatives is characterized in that may further comprise the steps:
(1) Toadpoison Medicine is mixed with mol ratio 1:2 with oxygenant, be dissolved in the organic solvent, after the reaction, silica gel column chromatography separates under the room temperature, obtains the intermediate B ufalinone of 3 oxidations of Toadpoison Medicine;
(2) oxidation products Bufalinone is mixed with mol ratio 1:2.5 with the organic amine salt hydrochlorate, be dissolved in the organic solvent, add alkaline reagents, react under the room temperature condition, obtain oxime formula intermediate;
(3) reductive agent of 3 times of oxime formula intermediate molar weights of adding in oxime formula intermediate is used organic solvent dissolution then, reacts under the ice-water bath condition, and silica gel column chromatography separates, and obtains the aglycon isomer of 3 α of Toadpoison Medicine and two kinds of configurations of β;
(4) the two kinds of aglycon isomer and the reducing sugar that respectively step (3) are obtained place organic solvent to react, and separate the Toadpoison Medicine glycosylated derivative that obtains suc as formula structure shown in I or the formula II through high performance liquid phase.
4. the synthetic method of Toadpoison Medicine glycosylated derivative according to claim 3 is characterized in that:
The described oxygenant of step (1) is the pyridinium chlorochromate hydrochloride, and organic solvent is CH
2Cl
2, reaction times 2.5h;
The described silica gel column chromatography of step (1) separates, and is the cyclohexane/ethyl acetate mixing solutions gradient elution with volume ratio 1:1.
5. the synthetic method of Toadpoison Medicine glycosylated derivative according to claim 3, it is characterized in that: the described organic amine salt hydrochlorate of step (2) is methoxamine hydrochloride, and organic solvent is methyl alcohol, and alkaline reagents is pyridine, and the reaction times is 2.5-3h.
6. the synthetic method of Toadpoison Medicine glycosylated derivative according to claim 3 is characterized in that:
The described reductive agent of step (3) is TERTIARY BUTYL AMINE base borane hydrochloride mixture, and organic solvent is that volume ratio is dioxane/alcohol mixeding liquid of 2:1, reaction times 3.5h;
The described silica gel column chromatography of step (3) separates, and is the cyclohexane/ethyl acetate mixing solutions gradient elution with volume ratio 4:1.
7. the synthetic method of Toadpoison Medicine glycosylated derivative according to claim 3 is characterized in that:
The described reducing sugar of step (4) is glucose, Fucose, wood sugar, lyxose or pectinose;
The described organic solvent of step (4) is that volume ratio is dimethyl formamide/alcohol mixeding liquid of 3:1, and the reaction times is 48h.
8. claim 1 or the 2 described Toadpoison Medicine glycosylated derivatives application in the preparation antitumor drug.
9. the application of Toadpoison Medicine glycosylated derivative according to claim 8 in the preparation antitumor drug is characterized in that: described antitumor drug is prevention or treatment prostate cancer, lung cancer, cancer of the stomach, liver cancer, mammary cancer, colorectal carcinoma, cervical cancer, skin carcinoma, nasopharyngeal carcinoma, oral carcinoma or leukemic medicine.
10. the application of Toadpoison Medicine glycosylated derivative according to claim 8 in the preparation antitumor drug, it is characterized in that: the formulation of described antitumor drug is tablet, injection, suppository, aerosol or nanometer formulation.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105030804A (en) * | 2015-07-14 | 2015-11-11 | 暨南大学 | Application of Bufalin-3beta-N-methoxyl-N-beta-D-heteroside in preparing cardiotonic drug |
| WO2018102973A1 (en) * | 2016-12-06 | 2018-06-14 | 江苏大学 | Pegylated vitamin e periplocymarin conjugate nanoparticles and preparation method therefor |
| CN111875660A (en) * | 2020-08-26 | 2020-11-03 | 四川大学 | Ouabain 1-position secondary hydroxyl derivative and preparation method and application thereof |
| CN111961106A (en) * | 2020-08-26 | 2020-11-20 | 四川大学 | Ouabain sugar ring 3' position hydroxyl derivative and its preparation method and use |
| CN112920253A (en) * | 2019-12-05 | 2021-06-08 | 中国科学院上海药物研究所 | Bufonis venenum diene derivative, and preparation method, application and pharmaceutical composition thereof |
-
2013
- 2013-06-04 CN CN201310217520.3A patent/CN103288911B/en active Active
Non-Patent Citations (4)
| Title |
|---|
| JOSEPH M. LANGENHAN ET AL.: "Enhancing the anticancer properties of cardiac glycosides by neoglycorandomization", 《PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA》 * |
| 邱峰: "《天然药物化学》", 30 September 2011 * |
| 陈大朋等: "强心苷类抗癌作用研究进展", 《中国药理学通报》 * |
| 陈少萍: "强心苷的抗肿瘤治疗研究进展", 《药学服务与研究》 * |
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| CN105030804A (en) * | 2015-07-14 | 2015-11-11 | 暨南大学 | Application of Bufalin-3beta-N-methoxyl-N-beta-D-heteroside in preparing cardiotonic drug |
| CN105030804B (en) * | 2015-07-14 | 2018-10-09 | 暨南大学 | - 3 β-N- methoxyl groups-N- β of Bufalin-application of the D-Glucose glycosides in preparing cardiac drug |
| WO2018102973A1 (en) * | 2016-12-06 | 2018-06-14 | 江苏大学 | Pegylated vitamin e periplocymarin conjugate nanoparticles and preparation method therefor |
| CN112920253A (en) * | 2019-12-05 | 2021-06-08 | 中国科学院上海药物研究所 | Bufonis venenum diene derivative, and preparation method, application and pharmaceutical composition thereof |
| CN111875660A (en) * | 2020-08-26 | 2020-11-03 | 四川大学 | Ouabain 1-position secondary hydroxyl derivative and preparation method and application thereof |
| CN111961106A (en) * | 2020-08-26 | 2020-11-20 | 四川大学 | Ouabain sugar ring 3' position hydroxyl derivative and its preparation method and use |
| CN111961106B (en) * | 2020-08-26 | 2022-05-20 | 四川大学 | Ouabain sugar ring 3' hydroxyl derivative, preparation method and application thereof |
| CN111875660B (en) * | 2020-08-26 | 2022-05-20 | 四川大学 | Ouabain 1-position secondary hydroxyl derivative and preparation method and application thereof |
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