CN101906102A - Beta-carboline alkaloid derivative, preparation method and application thereof - Google Patents
Beta-carboline alkaloid derivative, preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a beta-carboline alkaloid derivative with an anti-tumor activity, a preparation method and application thereof. In the invention, 3S-1-(2-carbonylethyl)-beta-carboline-3-benzyl ester carboxylate and benzyl ester amino acid are condensed or the 3S-1-(2-carbonylethyl)-beta-carboline-3-benzyl ester carboxylate and dibenzyl ester glutamic acid are condensed to obtain the derivative, wherein the benzyl ester amino acid is selected from glycine residues, alanine residues, valine residues, leucine residues, isoleucine residues, methionine residues, proline residues, phenylalanine residues, tryptophan residues, serine residues, threonine residues, asparagine residues, glutamine residues, tyrosine residues, benzyl aspartate residues, N omega-benzoxycarbonyl lysine residues and histidine residues. Proved by pharmacological experiments, the derivative has good anti-tumor cell proliferation activity and can be clinically used as an anti-tumor agent.
Description
Technical field
The present invention relates to alcaloid-derivatives, relate in particular to beta-carboline alkaloid derivative that has anti-tumor activity and preparation method thereof, the invention still further relates to this beta-carboline alkaloid derivative, belong to biomedicine field in application as antineoplastic agent.
Background technology
Malignant tumour is a kind of common disease and frequently-occurring disease of serious threat human health, and the mortality ratio that the mankind cause because of malignant tumour is second of all disease death rates, is only second to cardiovascular and cerebrovascular diseases.The tumor treatment method has operative treatment, radiotherapy and pharmacological agent (chemotherapy).At present, chemotherapy remains the main means of clinical treatment tumour.Seeking antitumor drug is one of focus of new drug research.The β-Ka Lin compounds is the natural indole alkaloid of a class, is distributed in nature widely.This compounds has anti-tumor activity and has caused extensive concern.The contriver recognizes, introduces benzyls and introduce amino-acid benzyl ester at 1 to produce antitumor action 3 of β-Ka Lin-3-carboxylic acid.According to this conception, the contriver proposes the present invention.
Summary of the invention
One of purpose of the present invention provides the beta-carboline alkaloid derivative that a class has anti-tumor activity;
Two of purpose of the present invention provides a kind of above-mentioned method with beta-carboline alkaloid derivative of anti-tumor activity for preparing;
Above-mentioned purpose of the present invention is achieved through the following technical solutions;
Beta-carboline alkaloid derivative with anti-tumor activity, its structural formula are shown in general formula I or the II:
Wherein, AA is a glycine residue, alanine residue, the Xie Ansuan residue, leucine residue, Isoleucine residue, methionine residue, proline residue, phenylalanine residue, tryptophan residue, serine residue, threonine residues, asparagine residue, glutamine residue, tyrosine residues, aspartic acid benzyl ester residue, N
ωSweet lysine residue of-carbobenzoxy-(Cbz) or histidine residues; R is a L-glutamic acid lactan residue.
A kind of method for preparing above-mentioned compound of Formula I may further comprise the steps:
(1) preparation 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate;
(2) with 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate and amino-acid benzyl ester condensation, promptly; Wherein, described amino-acid benzyl ester is selected from glycine benzyl ester, alanine benzyl ester, Xie Ansuan benzyl ester, leucine benzyl ester, Isoleucine benzyl ester, methionine(Met) benzyl ester, proline(Pro) benzyl ester, phenylalanine benzyl ester, tryptophan benzyl ester, Serine benzyl ester, threonine benzyl ester, l-asparagine benzyl ester, glutamine benzyl ester, tyrosine benzyl ester, aspartic acid benzyl ester, N
ω-carbobenzoxy-(Cbz) sweet Methionin benzyl ester or Histidine benzyl ester;
A kind of method for preparing above-mentioned general formula I I compound may further comprise the steps:
(1) in the presence of polyphosphoric acid and benzylalcohol, the L-glutamic acid rotating become ammonia propylhomoserin dibenzyl ester;
(2) preparation 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate;
(3) with 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate and the condensation of L-glutamic acid dibenzyl ester, promptly.
Wherein, described 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate can prepare with reference to following method:
(1) at SOCl
2Existence becomes the L-tryptophan benzyl ester with the L-tryptophan transfer down with methyl alcohol;
(2) in the presence of concentrated hydrochloric acid with L-tryptophan benzyl ester and 1,1,3,3-tetramethyl Ethylene Oxide condensation prepared 3S-1-(2, the 2-dimethoxy ethyl)-tetrahydrochysene-β-Ka Lin-3-benzyl carboxylate;
(3) at KMnO
4Exist down 3S-1-(2, the 2-dimethoxy ethyl)-tetrahydrochysene-β-Ka Lin-3-benzyl carboxylate is oxidized to 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate;
(4) in the MeOH of NaOH solution with 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate saponification, the preparation 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-carboxylic acid
(5) at K
2CO
3Use BrCH under existing
2C
6H
53 carboxyl esterifications of 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-carboxylic acid are generated 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate;
(6) at HOAc, HCl and H
2The O existence is hydrolyzed to 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate with 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate.
At tumor models and mouse S
180Estimate the anti-tumor activity of The compounds of this invention on the sarcoma model, test-results shows that The compounds of this invention has outstanding anti-tumor activity, can be used as potential antineoplastic agent clinically.
Another purpose of the present invention provides the pharmaceutical composition of the above-mentioned general formula compound of a kind of the present invention of containing, and this pharmaceutical composition is gone up effective dose by treatment The compounds of this invention is with pharmaceutically acceptable excipient or assist and add agent and form; That is: with the The compounds of this invention of significant quantity with after pharmaceutically acceptable carrier or thinner cooperate, by the formulation method of this area routine it is prepared into any one appropriate drug composition.Usually said composition is suitable for oral administration and drug administration by injection, also is fit to other medication.Said composition can be liquid preparation forms such as tablet, capsule, pulvis, granule, lozenge, suppository, or oral liquid.According to different medications, pharmaceutical composition of the present invention can contain 0.1%-99% weight, the The compounds of this invention of preferred 10-6O% weight.
Description of drawings
The synthetic route chart of Fig. 1 compound of Formula I of the present invention; I) SOCl
2, MeOH; Ii) HCl, 1,1,3,3-tetramethyl Ethylene Oxide, MeOH; Iii) KMnO
4, DMF; Iv) NaOH, H
2O, MeOH; V) K
2CO
3, BrCH
2C
6H
5, DMF/THF; Vi) HCl, HOAc, H
2O; Vii) NaOH, NaBH
3CN and glycine benzyl ester, L-alanine benzyl ester, L-Xie Ansuan benzyl ester, L-leucine benzyl ester, Isoleucine benzyl ester, the L-tryptophan benzyl ester, L-Serine benzyl ester, L-threonine benzyl ester, altheine benzyl ester, L-glutaminate benzyl ester, L-tyrosine benzyl ester, L-aspartic acid dibenzyl ester, L-N
ω-carbobenzoxy-(Cbz) Methionin benzyl ester, L-Histidine benzyl ester.AA is a glycine residue in 6a, and AA is an alanine residue among the 6b, and AA is the Xie Ansuan residue among the 6c, and AA is a leucine residue among the 6d, and AA is the Isoleucine residue among the 6e, R=CH among the 6f
2CH
2SCH
3, R=NH=NCH among the 6g
2CH
2CH
2, R=CH among the 6h
2C
6H
5, AA is a tryptophan residue among the 6i, and AA is a serine residue among the 6j, and AA is a threonine residues among the 6k, and AA is an asparagine residue among the 6l, and AA is a glutamine residue among the 6m, and AA is a tyrosine residues among the 6n, and AA is an aspartic acid benzyl ester residue among the 6o, AA is N among the 6p
ωThe sweet lysine residue of-carbobenzoxy-(Cbz), AA is a histidine residues among the 6q.
The synthetic route chart of Fig. 2 general formula I I of the present invention compound.
Embodiment
In order further to set forth the present invention, provide a series of embodiment below.These embodiment are illustrative fully, and they only are used for the present invention is specifically described, and not should be understood to limitation of the present invention.
Method is led in the preparation of embodiment 1L-amino-acid benzyl ester hydrochloride
With polyphosphoric acid (10g, 29.60mmol) with the 50ml phenylcarbinol after the dissolving of 90 ℃ of heating stirrers, add L-amino acid 20mmol.Reacted 8 hours.TLC shows no amino acid residue, and stopped reaction is cooled to room temperature.The aqueous sulfuric acid of adding 10%, and add the 20ml ether, separatory, the sodium bicarbonate aqueous solution of water 5% is adjusted pH=10.Extracted with diethyl ether three times (20ml * 3) merges organic phase, anhydrous sodium sulfate drying.Filter, feed HCl gas in the filtrate, getting L-amino-acid benzyl ester hydrochloride is white solid, and yield is between 23-72%.
Logical according to this method has been synthesized HClGly-OBzl, HClAla-OBzll, HClVal-OBzl, HClLeu-OBzl HClIle-OBzl, HClMet-OBzl, HClPro-OBzl, HClPhe-OBzl, HClTrp-OBzl, HClSer-OBzl, HClThr-OBzl, HClAsn-OBzl, HClGln-OBzl, HClTyr-OBzl, the amino acid whose benzyl ester hydrochloride of 18 kinds of L-of HClAsp-OBzl, HClLys (Z)-OBzl, HClHis-OBzl, HClGlu-OBzl.Physical constant such as fusing point, optically-active and the data consistent of having reported.
Embodiment 21-(2, the 2-dimethoxy ethyl)-2,3,4,9-tetrahydrochysene-β-Ka Lin benzyl carboxylate (2)
In the round-bottomed flask of 250ml, add L-tryptophan benzyl ester hydrochloride 5g (19.6mmol), 50ml methyl alcohol, 1,1,3,3-tetramethoxy propane 6ml (23.6mol) regulates pH to 2 with 5N hydrochloric acid, be warmed up to 45 ℃ under stirring, insulation reaction 48 hours, TLC monitoring raw material spot disappears, and reaction solution is cooled to room temperature, drip triethylamine and regulate pH greater than 8, decompression and solvent recovery adds 50ml trichloromethane and 50ml10% aqueous sodium carbonate in the resistates, water layer chloroform extraction (30ml * 3) behind the separatory, merge organic layer, anhydrous sodium sulfate drying filters, and filtrate decompression is concentrated into dried, resistates gets title compound with purification by silica gel column chromatography, is yellow oil 5.4g (86%).ESI-MS：m/e(M
+)318
Embodiment 31-(2, the 2-dimethoxy ethyl)-3-carboline carboxylate benzyl ester (3)
In the round-bottomed flask of 250ml, add 4.3g (13.783mmol) compound (2), be dissolved among the 100mlDMF, be stirred to the solution clarification, then under the ice bath cooling, add potassium permanganate 3.04g (19.296mmol) in batches, after about half hour, all add, room temperature reaction 3 hours, TLC monitoring raw material spot disappears, in reaction flask, add 20ml ethanol, stir after 20 minutes, filter, after filter cake extracts with hot ethyl acetate, waste residue discards, after the extracting solution cooling, difference water, saturated common salt water washing, anhydrous sodium sulfate drying, reclaim solvent, get the pure product of target compound, 520mg.After filtrate is reclaimed solvent, after residuum dissolves with the 50ml trichloromethane, water, saturated common salt water washing respectively, anhydrous sodium sulfate drying reclaims solvent, and residuum gets white powder 3.47g with acetone recrystallization, and yield is 80.32%.m.p.：134.5-136℃。ESI/MS(m/e)315[M+H]
+。
Embodiment 41-(2, the 2-dimethoxy ethyl)-3-carboline carboxylate carboxylic acid (4)
Under the ice bath, in the round-bottomed flask of 250ml, add 1.59mmol 1-(2, the 2-dimethoxy ethyl)-3-carboline carboxylate benzyl ester, be dissolved in the 7ml methanol solution, stir to clarify, pH to 11. ice bath of adjusting reaction solution with the aqueous sodium hydroxide solution of 2N reacts 3.5h, TLC (CHCl down
3/ CH
3OH, 10: 1) monitoring reaction no 1-(2, the 2-dimethoxy ethyl)-3-carboline carboxylate benzyl ester residue.Use KHSO
4After solution is transferred pH to 7. reclaim under reduced pressure methyl alcohol, residuum KHSO
4Solution is transferred pH to 2, with ethyl acetate extraction (30ml * 3), merges organic phase, water, saturated common salt water washing respectively, and anhydrous sodium sulfate drying reclaims solvent, obtains light yellow oil, not purifiedly directly applies to next step reaction.
Embodiment 51-(2, the 2-dimethoxy ethyl)-3-carboline carboxylate benzyl carboxylate (5)
Add compound (4) 3.81g (12.7mmol) in the round-bottomed flask of 250ml, the anhydrous THF of 20ml dry DMF and 20ml is stirred to the solution clarification, adds K
2CO
32.6g (19mmol) stirring at room 4h slowly drips bromobenzyl 2.37g (13.97mmol), room temperature reaction 8h, filtering reacting liquid, decompression and solvent recovery.Residuum gets white solid powder 3.51g through purification by silica gel column chromatography, and yield is 70.7%.Mp℃；ESI/MS(m/e)391[M+H]
+。
Embodiment 6S-1-(2-carbonyl ethyl)-9-H-β-Ka Lin-3-benzyl carboxylate (6)
In the round-bottomed flask of 100ml, add compound (4) 3.2g (8.26mmol), 80ml acetic acid, 10ml water and 10ml hydrochloric acid, stirring at room 4h, TLC monitoring raw material spot disappears, add the 300ml frozen water, ice bath stirs the 10min. after-filtration down, filter cake water-5% sodium bicarbonate aqueous solution-water washing dries, and is applied directly to next step reaction.
The synthetic logical method (6a-r) of embodiment 7N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethylamino acid benzyl ester
In the round-bottomed flask of 100ml, add 8.7mmol amino-acid benzyl ester hydrochloride, 2.4mmol sodium hydroxide, 25ml methyl alcohol, stirring at room 30min, add 5.79mmol compound 3S-1-(2-carbonyl ethyl)-9-H-β-Ka Lin-3-benzyl carboxylate (6) then, behind the 10min, add the 2.9mmol sodium cyanoborohydride, stirring at room 19h, TLC monitoring raw material spot disappears, add 3N hydrochloric acid and regulate pH to 2, decompression and solvent recovery adds 50ml water and 50ml acetic acid ethyl dissolution in the residuum, regulate pH to 8 with the 2N aqueous sodium hydroxide solution, separatory, ethyl acetate extraction (50ml * 3) merges organic phase, drying, decompression and solvent recovery, residuum gets title compound through purification by silica gel column chromatography.
Embodiment 8N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl glycine benzyl ester (6a)
Yield: 77.20%; ESI/MS (m/e) 494[M+H]
+
Embodiment 9N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl alanine benzyl ester (6b)
Yield: 68.02%.[α]
D 20=30.49 (c=0.01, CHCl
3), ESI/MS (m/e) 508[M+H]
+
Embodiment 10N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl Xie Ansuan benzyl ester (6c)
Yield: 64.95%.[α]
D 20=3.80 (c=0.01, CHCl
3), ESI/MS (m/e) 536[M+H]
+
Embodiment 11N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl leucine benzyl ester (6d)
Yield: 70.25%.[α]
D 20=-3.03 (c=0.01, CHCl
3), ESI/MS (m/e) 550[M+H]
+
Embodiment 12N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl Isoleucine benzyl ester (6e)
Yield: 65.32%.[α]
D 20=6.57 (c=0.01, CHCl
3), ESI/MS (m/e) 550[M+H]
+
Embodiment 13N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl methionine(Met) benzyl ester (6f)
Yield: 68.82%.[α]
D 20=-26.55 (c=0.01, DMSO), ESI/MS (m/e) 568[M+H]
+
Embodiment 14N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl proline(Pro) benzyl ester (6g)
Yield: 77.32%.[α]
D 20=-1.13 (c=0.01, DMSO), ESI/MS (m/e) 534[M+H]
+
Embodiment 15N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethylbenzene alanine benzyl ester (6h)
Yield: 70.29%.[α]
D 20=10.92 (c=0.01, CHCl
3); ESI/MS (m/e) 584[M+H]
+
Embodiment 16N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl tryptophan benzyl ester (6i)
Yield: 64.38%; [α]
D 20=1.13 (c=0.01, CHCl3); ESI/MS (m/e) 623[M+H]
+
Embodiment 17N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl Serine benzyl ester (6j)
Yield: 55.67%; [α]
D 20=-7.89 (c=0.01, CHCl
3); ESI/MS (m/e) 524[M+H]
+
Embodiment 18N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl threonine benzyl ester (6k)
Yield: 60.39%; [α]
D 20=-11.02 (c=0.01, CHCl
3); ESI/MS (m/e) 538[M+H]
+
Embodiment 19N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl asparagine benzyl ester (6l)
Yield: 50.28%.[α]
D 20=-31.14 (c=0.01, CHCl
3); ESI/MS (m/e) 551[M+H]
+
Embodiment 20N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethylglutamine benzyl ester (6m)
Yield: 67.5%; [α]
D 20=-5.8 (c=0.01, CHCl
3); ESI/MS (m/e) 474[M+H]
+
Embodiment 21N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl tyrosine benzyl ester (6n)
Yield: 92.76%.[α]
D 20=8.83 (c=0.01, CHCl
3); ESI/MS (m/e) 600[M+H]
+
Embodiment 22N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl aspartic acid benzyl ester (6o)
Yield: 49.73%.[α]
D 20=-7.96 (c=0.01, CHCl
3); ESI/MS (m/e) 642[M+H]
+
Embodiment 23N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl-N '-(carbobenzoxy-(Cbz)) Methionin benzyl ester (6p)
Yield：70.28％；[α]
D 20＝2.53(c＝0.01，CHCl
3)；ESI/MS(m/e)699[M+H]
+。
Embodiment 24N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl Histidine benzyl carboxylate (6q)
Yield: 42.29%; [α]
D 20=-7.99 (c=0.01, CHCl
3); ESI/MS (m/e) 574[M+H]
+
Embodiment 25N-(3-carbobenzoxy-(Cbz)-9-H-β-Ka Lin-1-yl) ethyl 5-oxo-pyrrolidine-2-benzyl carboxylate (6r)
Yield: 56.96%.[α]
D 20=-30.05 (c=0.01, DMSO), ESI/MS (m/e) 548[M+H]
+
Experimental example 1 The compounds of this invention is to the antiproliferative activity evaluation experimental of tumour cell
1) given the test agent
2) human cancer cell strain
Hela (cervical cancer cell of epithelial origin), HepG
2(hepatocellular carcinoma cells), Bel7402 (hepatocellular carcinoma cells), K562 (human leukemia cell), B16 (melanoma cell), Eahy926 (fusion cell line of human lung adenocarcinoma cell line A549 and Human umbilical vein endothelial cells hybridization).
3) key instrument
Microplate reader: 450, Biorad company
High-pressure sterilizing pot: 400Ep-Z, Bruckmanning company
Cell incubator: INC153, memmer company
Refrigerated centrifuge: SPD111V, Thermo company
96 porocyte culture plates: Costar company
Quartzy automatic dual pure water distiller: 1810-B, Jiangsu high honour instrument Manufacturing Co., Ltd
4) main agents
MTT: four tetrazolium bromides (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl-tetrazolium bromide) (Flka), is dissolved among the PBS, and concentration is 5mg/ml, filtration sterilization, and 4 ℃ keep in Dark Place.
PBS: contain NaCl 8.2g in every liter of solution, KCl 0.20g, Na
2HPO
4.H
2O1.56g, KH
2PO
40.2g pH is 7.4
RPMI-1640 substratum: Gibco company
DMEM substratum: Gibco company
Mccy ' the s 5A medium+L-Glutamine substratum of improvement: Gibco company
Foetal calf serum: Hyclone company
Penicillin: Zhongnuo Pharmaceutical (Shijiazhuang) Co., Ltd., Shiyao Group
Streptomycin sulphate: Zhongnuo Pharmaceutical (Shijiazhuang) Co., Ltd., Shiyao Group
DMSO:Hyclone company.
All the other agents useful for same are commercially available analytical pure.
5) experimental technique.
Respectively that growth conditions is good, be in the tumour cell of logarithmic phase with 5 * 10
4The density of individual/mL is inoculated in 96 orifice plates, every hole 100 μ l, by default concentration gradient add to be measured, through the sample of sterilising treatment, control group adds the solvent of isopyknic sample dissolution.Continue to cultivate after 48 hours, it is the MTT solution of 5mg/ml that every hole adds 25 μ l concentration, continue to place 37 ℃ to hatch four hours, nutrient solution in the careful sucking-off hole, every hole adds the DMSO dissolve purple residue (first a ceremonial jade-ladle, used in libation) of 100 μ l, about 15min crystallisate that vibrates all dissolves, and measures each hole O.D. value (absorption value) on the 570nm microplate reader.
Calculate sample under each sample concentration to the inhibiting rate of tumour cell according to formula " relative survival rate=(D pastille-D blank)/(D contrast-D blank) * 100% ".
Experiment repeats 3 times, to draw the cell viability curve, obtains IC
50Value.
6) experimental result
In cell in vitro poison examination experiment, estimated the effect of The compounds of this invention 6a-r to six kinds of human tumour cell lines' such as HepG2, Hela, Bel7402, K562, eahy926, B16 competence for added value, experimental result sees Table 1.
Table 1 The compounds of this invention 6a-r extracorporeal anti-tumor cell-proliferation activity (IC
50± SD, μ M)
| Compd a | HepG 2 | Hela | Bel7402 | | eahy926 | B16 | |
| 6a | 50.0±8.3 | 26.8±4.2 | 53.2±7.6 | 47.1±6.2 | 28.7±3.7 | 75.4±9.1 | |
| 6b | ＞100 | 76.4±1.2 | 100±5.2 | 48.2±8.4 | 4.6±1.3 | 20.5±2.3 | |
| 6c | 29.2±1.2 | 25.5±2.2 | 20.3±3.1 | 20.9±1.8 | 15.0±1.4 | 28.8±1.3 | |
| 6d | 25.2±2.1 | 22.5±3.1 | 43.5±1.4 | 21.5±1.8 | 17.8±2.1 | 50.2±3.2 | |
| 6e | ＞100 | ＞100 | ＞100 | ＞100 | ＞100 | 19.2±2.1 | |
| 6f | 18.7±1.2 | 28.8±1.5 | 25.8±2.1 | 13.5±1.0 | 18.5±2.9 | 61.1±2.8 | |
| 6g | 17.12±1.2 | 25.81±1.5 | 52.81±2.2 | 20.12±1.9 | 26.11±1.7 | 71.83±2.2 | |
| 6h | ＞100 | ＞100 | ＞100 | ＞100 | 23.0±2.1 | 28.1±2.1 | |
| 6i | 60.0±3.2 | 59.5±2.7 | 39.0±2.1 | 14.9±1.2 | 34.9±4.7 | 20.7±1.3 |
| 6j | 19.9±1.1 | 20.3±2.0 | 54.6±6.8 | 19.7±2.1 | 13.2±2.3 | 60.3±8.4 |
| 6k | 26.0±2.1 | 15.2±1.2 | ＞100 | 38.3±3.2 | 37.2±1.9 | 38.4±3.3 |
| 6l | ＞100 | 97.2±15.2 | ＞100 | 63.6±10.3 | ＞100 | 17.1±4.5 |
| 6m | 35.9±5.2 | 71.0±8.2 | 81.3±6.7 | ＞100 | 96.1±9.8 | 42.5±6.2 |
| 6n | ＞100 | 16.7±2.0 | 23.7±2.3 | 18.9±1.9 | 16.6±2.3 | 24.5±2.8 |
| 6o | ＞100 | ＞100 | 60.3± | ＞100 | 69.5±8.7 | 11.1±4.5 |
| 6p | 36.28±4.2 | 33.85±4.8 | 14.91±3.2 | 15.37±2.8 | 10.26±1.7 | 49.4±5.4 |
| 6q | ＞100 | 63.56±7.2 | 36.85±5.3 | 14.43±3.1 | 26.17±2.8 | 60.02±4.6 |
| 6r | 63.09±10.2 | 54.94±8.2 | ＞100 | 18.53±4.5 | 96.1±11.2 | 66.37±5.2 |
| ADR | 0.19±0.1 | 34.18±2.1 | 33.45±1.8 | 27.22±2.3 |
The ADR=cytosine arabinoside
Experiment in vitro is the result show, The compounds of this invention 6a-r all has outstanding anti-tumor activity.
Experimental example 2 The compounds of this invention anti-tumor in vivo activity experiments
1) experiment material
The compound 6a-r that test-compound: embodiment is prepared.
Positive reference substance is a cytosine arabinoside.
Laboratory animal: Kunming mouse (KM cleans level), male, body weight
Provide by experimentation on animals center, Department Of Medicine, Peking University.One group of per 10 mouse, each one group of blank and positive control.
Knurl source: mouse S
180Sarcoma is provided by experimentation on animals center, Department Of Medicine, Peking University, goes down to posterity voluntarily and keeps.
Solvent: physiological saline, 0.5%CMC-Na solution.
2) dosage setting
Test-compound 6a-r be made as height (89 μ mol/kg), in (8.9 μ mol/kg), low (0.89 μ mol/kg) three dosage groups, all adopt the abdominal cavity single-dose.
3) medicine preparation
Test-compound 6a-r indissoluble in water adds the wetting hydrotropy of a spot of tween 80 during experiment, add 0.5%CMC-Na solution to required concentration gradually and get final product.The positive reference substance cytosine arabinoside is water-soluble, adopts physiological saline solution to get final product.
4) dosage and dosage regimen
Test-compound 6a-r is all with the abdominal cavity single-dose.By corresponding dosage once a day, the 0.2ml/ mouse, successive administration 7 days, administration is 7 times altogether.
Negative control is with isopyknic corresponding solution, all with intraperitoneal administration.By corresponding dosage once a day, the 0.2ml/ mouse, successive administration 7 days, administration is 7 times altogether.
The positive reference substance cytosine arabinoside is by the dosage of 89 μ mol/kg, intraperitoneal administration.Once a day, the 0.2ml/ mouse, successive administration 7 days, administration is 7 times altogether.
5) foundation of animal model
Adopt anti-tumor in vivo armpit subcutaneous vaccination model: under aseptic condition, extract inoculation 7d, after get the vigorous S of growth
180Ascitic tumor knurl liquid, be diluted to the liquid thorough mixing of (1: 2) with physiological saline, the tumour cell suspension is dyeed with freshly prepared 0.2% trypan blue, count by the white blood cell count(WBC) method behind the mixing, dye blue person and be dead cell, tinter is not a viable cell, is calculated as follows cell concn and cell survival rate.
Viable count/4 * 10 in the big grid in cell concn=4
4* extension rate=cell count/ml
Cell survival rate=viable count/(viable count+dead cell number) * 100%
Survival rate is prepared into 1 * 10 greater than 90% knurl liquid with the homogenate method
7The cell suspension of individual/ml in corresponding host's armpit subcutaneous vaccination 0.2ml/ mouse, is made the solid tumor animal model.
6) detect index and method
7) neurotoxicity is observed in the body
Observe autonomic activities, the mental status, hair, breathing, the diet of the reaction mouse of each treated animal of administration every day, the ight soil proterties.
8) mensuration of solid tumor tumour inhibiting rate and body weight gain
After each organizes successive administration 7d, take off cervical vertebra in 8d and put to death mouse, take by weighing body weight (execution body weight), with the fixing right armpit tumor location of mouse of tweezers, cut off skin then, the exposure tumour, blunt separation is weighed, and is calculated as follows tumour inhibiting rate.
The average knurl of the average knurl weight-administration of tumour inhibiting rate %=[(negative control group group is heavy)/the average knurl of negative control group is heavy] * 100%.
Body weight gain (g)=execution body weight-original body weight-knurl is heavy.
10) statistical method
This experimental data statistics all adopts t check and variance analysis, with
Expression.
11) experimental result
Compound 6a-r is to lotus S
180The anti-tumor in vivo activity experiment result of sarcoma is as shown in table 2.Dosage is the compound 6a-r administration group of 8.9 μ mol/kg, and after 7 days, average tumor weight is from 0.71 ± 0.18g to 1.70 ± 0.55g through continuous intraperitoneal administration; The average tumor weight of negative control group is 1.62 ± 0.28, there are 12 compounds to have remarkable antitumor effect among the compound 6a-r, compound 6b wherein, i, l, m (the heavy scope of knurl is 0.74 ± 0.09g to 0.85 ± 0.21g, compares p＞0.05 with the cytosine arabinoside positive controls) anti-tumor activity is suitable with cytosine arabinoside.Compound 6d, e, h, p (the heavy scope of knurl is 1.08 ± 0.15g to 0.94 ± 0.16g, compares p＜0.05 with the physiological saline group) anti-tumor activity and blank group have significant difference.The anti-tumor activity of finding this type compound from analysis of experimental data is relevant with the substituent character of the 1-of R, for example, when the R group be that the little fat group (methyl or sec.-propyl) or the compound of acidic-group (carboxymethyl, phenylol) or basic group (guanidine radicals and indyl) have goodish anti-tumor in vivo activity.
Table 2 The compounds of this invention 6a-p is to lotus S
180The influence of mouse tumor weight
a
Arc (cytosine arabinoside and 6a-r dosage are 8.9 μ mol/kg, the NS=solvent, and n=12, average knurl heavily is expressed as
B) compare p＜0.05 with NS, c) compare p＜0.01, compare p＞0.05 with cytosine arabinoside with NS; D) compare p＜0.01 with NS, compare p＜0.05 with cytosine arabinoside; E) compare its p＜0.01 with NS, cytosine arabinoside.
Compound 6b, j, q are the active ideal compound of anti-tumor in vivo among the The compounds of this invention 6a-p.They strengthen (table 3) to the restraining effect dose-dependently ground of tumour when 0.89,8.9 and 89.0 μ mol/kg dosage.
Table 3 various dose 6b, j, q is to lotus lotus S
180The influence of mouse tumor weight
a
A NS=solvent, n=12, average knurl heavily is expressed as
Claims (8)
1. the beta-carboline alkaloid derivative that has anti-tumor activity, its structural formula are shown in general formula I or the II:
Wherein, AA is a glycine residue, alanine residue, the Xie Ansuan residue, leucine residue, Isoleucine residue, methionine residue, proline residue, phenylalanine residue, tryptophan residue, serine residue, threonine residues, asparagine residue, glutamine residue, tyrosine residues, aspartic acid benzyl ester residue, N
ωSweet lysine residue of-carbobenzoxy-(Cbz) or histidine residues; R is a L-glutamic acid lactan residue.
2. method for preparing the described compound of Formula I of claim 1 may further comprise the steps:
(1) preparation 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate;
(2) with 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate and amino-acid benzyl ester condensation, promptly; Wherein, described amino-acid benzyl ester is selected from glycine benzyl ester, alanine benzyl ester, Xie Ansuan benzyl ester, leucine benzyl ester, Isoleucine benzyl ester, methionine(Met) benzyl ester, proline(Pro) benzyl ester, phenylalanine benzyl ester, tryptophan benzyl ester, Serine benzyl ester, threonine benzyl ester, l-asparagine benzyl ester, glutamine benzyl ester, tyrosine benzyl ester, aspartic acid benzyl ester, N
ω-carbobenzoxy-(Cbz) sweet Methionin benzyl ester or Histidine benzyl ester.
3. it is characterized in that in accordance with the method for claim 2: at NaOH and NaBH
3CN exists down 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate and amino-acid benzyl ester condensation.
4. method for preparing the described general formula I I of claim 1 compound may further comprise the steps:
(1) in the presence of polyphosphoric acid and benzylalcohol, the L-glutamic acid rotating become ammonia propylhomoserin dibenzyl ester;
(2) preparation 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate;
(3) with 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate and the condensation of L-glutamic acid dibenzyl ester, promptly.
5. it is characterized in that in accordance with the method for claim 4: at NaOH and NaBH
3CN exists down 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate and the condensation of L-glutamic acid dibenzyl ester.
6. according to claim 2 or 4 described methods, it is characterized in that described 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate prepares in accordance with the following methods:
(1) at SOCl
2Existence becomes the L-tryptophan benzyl ester with the L-tryptophan transfer down with methyl alcohol;
(2) in the presence of concentrated hydrochloric acid with L-tryptophan benzyl ester and 1,1,3,3-tetramethyl Ethylene Oxide condensation prepared 3S-1-(2, the 2-dimethoxy ethyl)-tetrahydrochysene-β-Ka Lin-3-benzyl carboxylate;
(3) at KMnO
4Exist down 3S-1-(2, the 2-dimethoxy ethyl)-tetrahydrochysene-β-Ka Lin-3-benzyl carboxylate is oxidized to 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate;
(4) in the MeOH of NaOH solution with 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate saponification, the preparation 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-carboxylic acid;
(5) at K
2CO
3Use BrCH under existing
2C
6H
53 carboxyl esterifications of 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-carboxylic acid are generated 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate;
(6) at HOAc, HCl and H
2O is hydrolyzed to 3S-1-(2-carbonyl ethyl)-β-Ka Lin-3-benzyl carboxylate with 3S-1-(2, the 2-dimethoxy ethyl)-β-Ka Lin-3-benzyl carboxylate under existing.
7. pharmaceutical composition for the treatment of tumour is made up of the described beta-carboline alkaloid derivative of the claim 1 of significant quantity and pharmaceutically acceptable carrier or auxiliary material.
8. the described beta-carboline alkaloid derivative of claim 1 is in the purposes of preparation in the antitumor drug.
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