CN104231042A - 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, antitumor activity and application thereof - Google Patents

9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, antitumor activity and application thereof Download PDF

Info

Publication number
CN104231042A
CN104231042A CN201310225711.4A CN201310225711A CN104231042A CN 104231042 A CN104231042 A CN 104231042A CN 201310225711 A CN201310225711 A CN 201310225711A CN 104231042 A CN104231042 A CN 104231042A
Authority
CN
China
Prior art keywords
carboline
amino acid
obzl
beta
acid benzyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201310225711.4A
Other languages
Chinese (zh)
Inventor
赵明
彭师奇
王玉记
吴建辉
张攀
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Capital Medical University
Original Assignee
Capital Medical University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Capital Medical University filed Critical Capital Medical University
Priority to CN201310225711.4A priority Critical patent/CN104231042A/en
Publication of CN104231042A publication Critical patent/CN104231042A/en
Pending legal-status Critical Current

Links

Landscapes

  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, an antitumor activity and an application thereof. The invention discloses 16 kinds of 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters represented by the formula I, wherein in the formula I, AA is selected from the group consisting of L-Leu, L-Glu, L-Val, L-Trp, L-Arg, L-Tyr, L-Met, L-Phe, L-Ala, L-Gly, L-His, L-Asp, L-Ile, L-Lys, L-Ser and L-Thr residues. The invention discloses the preparation method of the 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters. The invention discloses an inhibitory effect of the 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters on proliferation of HL60 (human promyelocytic leukemia cells), K562 (human chronic myelocytic leukemia cells), HeLa (epithelial-derived cervical cancer cells), HepG2 (hepatocellular carcinoma cells) and HT-29 (human colon cancer cells). The invention further discloses an inhibitory effect of the 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters on tumor growth of S180-burdened mice. Therefore, the invention discloses clinical application prospects of the 16 kinds of 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters represented by the formula I as antitumor drugs.

Description

9-methoxyl methyl-β-carboline-3-formyl amino acid formyl amino acid benzyl ester, its synthesis, anti-tumor activity and application
Invention field
The present invention relates to 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester, relate to their preparation, relate to them to K562, HL60, Hela, HepG2, the inhibit activities of HT-29 five strain tumor cell proliferation, and relate to their restraining effect to mice bearing S180 tumor propagation further.Thus the present invention relates to the potential applicability in clinical practice of 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester as antitumor drug.The invention belongs to biomedicine field.
Background technology
Tumour be body under the effect of various tumorigenesis factor, the cell of local organization loses the regulation and control to growth at gene level, causes monoclonicity paraplasm and the true tumor that formed.Worldwide, tumour serious harm human health, affects human life quality, becomes one of topmost lethal cause of disease.One of important sources of natural product anti-cancer agent.β-carboline is from Herba pegani harmalae, be separated the antineoplastic compound obtained, and kills tumour cell in cytotoxic mode.This mode of action makes the toxicity of β-carboline large.Modify the structure of β-carboline to reduce poison and to improve anti-tumor activity, attracted to study interest evaluation widely.Contriver's understanding in the structural modification of β-carboline, introduces carboxyl and generates the 3-formyl amino acid formyl amino acid benzyl ester of β-carboline further with amino-acid benzyl ester coupling for 3 at β-carboline, tentatively just can fall the low toxicity of β-carboline and improve the target of anti-tumor activity.Contriver recognizes further, introduces 9-methoxyl methyl again can reach the low toxicity falling β-carboline and the target improving anti-tumor activity further 9 of 3-formyl amino acid formyl amino acid benzyl ester of β-carboline.On the basis of these understanding of contriver, inventors herein propose and 16 kinds of inventions that 9-methoxyl methyl-β-carboline-3-formyl amino acid formyl amino acid benzyl ester is relevant.The understanding of the contriver just spoken of, makes the present invention show two outstanding creativeness compared with the invention mentioned before contriver: the event resolves 1) killing tumour cell in cytotoxic mode; 2) activity of Tumor suppression growth increases substantially.
Summary of the invention
First content of the present invention is the 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester of design formula 1.
Second content of the present invention is to provide the preparation method of the 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester of formula 1, and the method comprises:
1) L-Trp is changed into 3S-9-methylol-tetrahydro-beta-carboline-3-carboxylic acid (1);
2) 3S-9-methylol-tetrahydro-beta-carboline-3-carboxylic acid (1) is changed into 3S-9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylate methyl ester (2);
3) make to change 9-methoxyl methyl-β-carboline-3-carboxylate methyl ester (3) into potassium permanganate oxidation 3S-9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylate methyl ester (2);
4) 9-methoxyl methyl-β-carboline-3-carboxylate methyl ester (3) is changed into 9-methoxyl methyl-β-carboline-3-carboxylic acid (4) in the basic conditions;
5) 9-methoxyl methyl-β-carboline-3-carboxylic acid (4) and 16 seed amino acid benzyl esters are obtained by reacting 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester (5a-p).
The method can describe by the synthetic route of the 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester of Fig. 1.
3rd content of the present invention evaluates the anti-tumour cell proliferative activity of 5a-p.Namely employing mtt assay evaluation 5a-p suppression K562, HL60, Hela, HepG2, HT-29 five strain proliferative activity o f tumor, calculates the IC that 5a-p suppresses these five kinds of tumor cell proliferations 50value.
4th content of the present invention evaluates 6a-p to the restraining effect of mice bearing S180 tumor propagation.
figure of description
The synthetic route .i of Fig. 1 .16 kind 9-methoxyl methyl-β-carboline-3-formyl amino acid formyl amino acid benzyl ester) dilute sulphuric acid, formaldehyde; Ii) methyl alcohol, thionyl chloride; Iii) DMF, potassium permanganate, ethanol; Iv) methyl alcohol, 1N NaOH, 1N HCl; V) .DCC, HOBt, NMM, DMF, amino-acid benzyl ester wherein AA=alanine residue in 5a; AA=Nitro-Arginine residue in 5b; AA=aspartic acid benzyl ester residue in 5c; AA=benzyl glutamate residue in 5d; AA=glycine residue in 5e; AA=histidine residues in 5f; AA=isoleucine residues in 5g; AA=leucine residue in 5h; AA=lysine residue in 5i; AA=methionine residues in 5j; AA=phenylalanine residue in 5k; AA=serine residue in 5l; AA=threonine residues in 5m; AA=tryptophan residue in 5n; AA=tyrosine residues in 5o; AA=valine residue in 5p.
Embodiment
In order to set forth the present invention further, provide a series of embodiment below.These embodiments are illustrative completely, and they are only used for being specifically described the present invention, not should be understood to limitation of the present invention.
Embodiment 1 prepares 3S-9-methylol-tetrahydro-beta-carboline-3-carboxylic acid (1)
In 800ml distilled water, add 6 98% vitriol oils, add 10g L-Trp, stir, make to dissolve completely, then add 40ml formalin, stir 48 hours, adjust pH7 with ammoniacal liquor, leave standstill 30min, have a large amount of colorless solid to settle out.Suction filtration, obtains 11g (90%) title compound, is colorless solid.ESI-MS(m/e)246[M-H] -。
Embodiment 2 prepares 3S-9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylate methyl ester (2)
In 120ml methyl alcohol, slowly drip 8ml thionyl chloride under ice bath stirs, dropwise rear continuation and stir 30min.In mixed solution, add 11g3S-9-methylol-tetrahydro-beta-carboline-3-carboxylic acid (1), stir 48 hours.Suction filtration, obtaining 6 (50%) title compounds, is faint yellow solid.ESI-MS(m/e)274[M-H] -。
Embodiment 3 prepares 9-methoxyl methyl-β-carboline-3-carboxylate methyl ester (3)
Be added to by 300ml DMF (DMF) in 5g3S-9-methyl-tetrahydro-β-carboline-3-carboxylate methyl ester (2), ice bath stirs, and slowly adds 6g potassium permanganate in batches.Stirring at room temperature 48 hours, inwardly adds 300ml ethanol, stirs 3 hours.Filter, filtrate reduced in volume, the red brown solid obtained adds dissolve with ethanol, elimination insolubles, the light yellow clear liquid obtained is evaporated to dry again, the yellow solid obtained uses column chromatography purification, and (sherwood oil: acetone=4: 1), 2.3g (45%) title compound are colorless solid.ESI-MS(m/e)270[M-H] -。
Embodiment 4 prepares 9-methoxyl methyl-β-carboline-3-carboxylic acid (4)
By 3.0g9-methoxyl methyl-β-carboline-3-carboxylate methyl ester (3) methyl alcohol to ultrasonic positive good dissolving.Stir, ice bath, drips NaOH solution (1mol/L) to pH=10 in reaction solution.Stirring reaction, uses TLC to monitor (methylene dichloride: methyl alcohol=15: the phosphor dot 1) to Rf=0.3 place disappears.Obtain light yellow clear liquid.Ice bath, inwardly drips HCl solution (1mol/L) to pH value of solution=7.White solid is separated out in orange-yellow clear solution.Filter, obtain white solid and be about 2g, productive rate 70%.ESI-MS(m/e)256[M-H] -
Embodiment 5 prepares 9-methoxyl methyl-β-carboline-3-formyl alanyl benzyl ester (5a)
Dissolved by 842mg (2.4mmol) ALANINE benzyl ester tosilate 15mL DMF (DMF), ice bath adds N-methylmorpholine (NMM) under stirring and adjusts pH10, stirs 30min, obtains A liquid.512mg 40mL DMF is dissolved, after cooling, adds 326mg HOBt, stir 10min.494mg DCC is dissolved in 5mL DMF and obtains B liquid.B liquid is added in A liquid, stirs 24h, TLC monitoring reaction and terminate.Reaction mixture is evaporated to dry.The residue with Ethyl acetate obtained dissolves, the solution obtained uses saturated sodium bicarbonate aqueous solution to wash (30mL × 3) successively, saturated sodium-chloride water solution is washed (30mL × 3), 5% aqueous potassium hydrogen sulfate is washed (30mL × 3), saturated sodium-chloride water solution is washed (30mL × 3), saturated sodium bicarbonate aqueous solution is washed (30mL × 3), and saturated sodium-chloride water solution is washed (30mL × 3).The ethyl acetate layer anhydrous sodium sulfate drying merged, filters, filtrate reduced in volume.With column chromatography purification, (sherwood oil: acetone=3: 1), obtains 0.450g (54%) title compound to residue, is colorless solid.Mp88-89℃。 ESI-MS(m/e)418[M+H] +。IR(KBr)：3373，1745，1652，1623，1558，1525，1492，1449，1386，1346，1203，1115，755，530，431。 1HNMR(300MHz，CDCl 3)8.9(d，J＝2.7Hz，2H)，8.64(d，J＝5.2Hz，1H)，8.20(d，J＝7.8Hz，1H)，7.65(d，J＝2.7Hz，2H)，7.40(m，6H)，5.79(s，2H)，5.26(s，2H)，4.94(m，1H)，3.27(s，3H)，1.64(d，J＝4.8Hz2H)。 13C-NMR(DMSO-d6，75MHz)：δ/ppm＝172.91，164.89，141.71，140.68，138.06，135.59，130.67，129.87，129.16，128.59，128.32，128.11，122.27，122.00，121.46，114.49，110.25，74.61，67.03，56.44，33.96，18.63。
Embodiment 6 prepares 9-methoxyl methyl-β-carboline-3-formyl Nitro-Arginine benzyl ester (5b)
According to the method for embodiment 5, obtaining 0.280mg (26%) title compound from 577mg (1.2mmol) L-NG-nitroarginine benzyl ester tosilate, is colorless solid.Mp78.7-79.5℃。 ESI-MS(m/e)548[M+H] +。IR(KBr)：3296，2951，1734，1652，1623，1557，1521，1495，1463，1419，1394，1251，1191，1151，1113，1052，1019，749，726。 1HNMR(300MHz，CDCl 3)8.9(d，J＝8.7Hz，2H)，8.80(d，J＝10.6Hz，2H)，8.23(d，J＝7.5Hz，1H)，7.65(m，2H)，7.21(s，6H)，5.76(s，2H)，5.21(s，2H)，4.94(m，1H)，3.79(s，1H)，3.27(s，3H)，2.23(s，2H)，2.18(s，1H)，1.90(s，1H)，1.75(s，2H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝171.94，166.36，159.43，141.68，139.54，138.17，135.07，130.99，129.91，129.33，128.73，128.67，128.45，122.64，121.84，121.63，114.88，110.22，74.55，67.60，56.46，50.80，40.38，30.95，24.53。
Embodiment 7 prepares the two benzyl ester (5c) of 9-methoxyl methyl-β-carboline-3-carbamyl aspartic acid
According to the method for embodiment 5, obtaining 857mg (52%) title compound from 873mg (1.8mmol) L-Aspartic acid benzyl ester tosilate, is clear yellow viscous solid. ESI-MS(m/e)548?[M+H] +。IR(KBr)：3032，2946，2890，2821，2358，1868，1800，1748，1733，1717，1699，1653，1647，1635，1623，1616，1558，1540，1521，1501，1496，1457，1336，1193，1114，750.418。 1HNMR(300MHz，CDCl 3)9.10(d，J＝8.9Hz，1H)，8.84(s，2H)，8.12(d，J＝7.8Hz，1H)，7.65(d，J＝6.3Hz2H)，7.21(d，J＝4.8Hz，11H)，5.76(s，2H)，5.32(m，2H)，5.24(d，J＝3.0Hz，2H)，5.15(m，1H)，3.32(d，J＝3.3Hz，1H)，3.27(s，3H)，3.14(m，1H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝171.04，170.89，164.94，141.94，140.54，138.25，136.29，136.24，132.17，129.60，129.22，128.83，128.46，128.46，128.45，128.36，128.11，127.07，122.98，121.64，121.61，114.79，111.55，74.33，66.84，66.36，56.24，49.29，36.40。
Embodiment 8 prepares the two benzyl ester (5d) of 9-methoxyl methyl-β-carboline-3-carbamylglutamic
According to the method for embodiment 5, obtaining 329mg (58%), target compound from the two benzyl ester tosilate of 647mg (1.3mmol) Pidolidone, is yellow solid. Mp78.7-79.5℃。ESI-MS(m/e)548[M+H] +。IR(KBr)：3033，2568，1733，1716，1683，1558，1539，1520，1507，1489，1457，1186，1154，920，587，431。 1HNMR(300MHz，CDCl 3)8.9(d，J＝8.7Hz，2H)，8.80(d，J＝10.6Hz，2H)，8.23(d，J＝7.5Hz，1H)，7.65(m，2H)，7.21(s，6H)，5.76(s，2H)，5.21(s，2H)，4.94(m，1H)，3.79(s，1H)，3.27(s，3H)，2.23(s，2H)，2.18(s，1H)，1.90(s，1H)，1.75(s，2H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝172.46，171.74，165.26，141.71，140.45，138.11，135.80，135.37，130.74，129.84，129.18，128.62，128.51，128.41，128.26，128.19，126.98，122.28，122.00，121.48，114.59，110.28，74.61，67.27，66.44，56.45，51.77，30.47，27.94。
Embodiment 9 prepares 9-methoxyl methyl-β-carboline-3-formylglycine benzyl ester (5e)
According to the method for embodiment 5, obtaining 240mg (59.5%) title compound from 404mg (1.2mmol) L-glycine benzyl ester p-toluene sulfonic acid salt, is colorless solid.Mp98-99℃。 IR(KBr)：3902，3750，3648，3378，3057，3033，2946，1742，1658，1624，1521，1465，1361，1344，1185，1158，1109，913，748，695ESI-MS(m/e)404[M+H] +1HNMR(300MHz，CDCl 3)8.9(d，J＝8.7Hz，2H)，8.89(s，1H)，8.25(d，J＝7.5Hz，1H)，7.69(m，2H)，7.41(m，6H)，5.83(s，2H)，5.27(s，2H)，4.41(d，J＝5.7Hz，1H)，3.35(s，3H) 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝169.44，135.28，128.64，128.43，123.24，110.9，77.43，77.2，77.01，76.59，75.54，67.44，56.96，41.99.Chemical?Formula：C 23H 21N 3O 4，Exact?Mass：403.15；Elemental?Analysis：C，68.47；H，5.25；N，10.42；O，15.86。
Embodiment 10 prepares 9-methoxyl methyl-β-carboline-3-formyl Histidine benzyl ester (5f)
According to the method for embodiment 5, obtaining 150mg (28%) title compound from 500mg (1.2mmol) L-Histidine benzyl ester tosilate, is light yellow solid.Mp83-84℃.。 ESI-MS(m/e)484[M+H] +。IR(KBr)：3852，3739，3648，3377，3057，2945，1742，1648，1521，1465，1339，11841158，1109，1061，1010，913，748，694。 1HNMR(300MHz，CDCl 3)8.94(d，J＝8.7Hz，2H)，8.20(d，J＝16.9Hz，2H)，8.17(d，J＝13.5Hz，2H)，7.64(s，4H)，7.40(m，2H)，7.29(s，1H)，5.78(s，4H)，3.82(s，1H)，3.66(d，J＝3.9Hz，2H)，3.35(s，3H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝169.96，144.95，141.65，136.87，136.74，130.34，129.75，129.23，129.13，122.16，121.76，121.71，121.33，121.22，116.56，115.82，110.18，74.54，71.54，61.42，56.46，36.19。
Embodiment 11 prepares 9-methoxyl methyl-β-carboline-3-formyl Isoleucine benzyl ester (5g)
According to the method for embodiment 5, obtaining 501mg (54%) title compound from 943mg (2.4mmol) ILE benzyl ester tosilate, is clear yellow viscous solid. ESI-MS(m/e)460[M+H] +。IR(KBr)：3853，3735，3649，3545，2964，2877，2359，1733，1683，1646，1558，1521，1507，1490，1457，1338，1045，1018，756，634，521。 1HNMR(300MHz，CDCl 3)8.9(d，J＝8.7Hz，2H)，8.66(d，J＝9.0Hz，1H)，8.22(d，J＝7.8Hz，1H)，7.65(d，J＝5.4Hz，2H)，7.41(m，6H)，5.83(s，2H)，5.24(m，2H)，4.94(m，1H)，3.35(s，3H)，2.15(m，1H)，1.58(m，1H)，1.40(m，1H)，1.03(d，J＝6.6Hz，3H)1.00(t，J＝14.7Hz，3H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝171.62，142.53，137.45，135.60，130.35，128.54，128.34，128.29，122.67，122.13，121.54，115.95，110.55，74.88，66.88，57.61，56.63，37.79，25.39，15.78，11.60。
Embodiment 12 prepares 9-methoxyl methyl-β-carboline-3-formyl leucine benzyl ester (5h)
According to the method for embodiment 5, from 943mg (2.4mmol) L-Leu benzyl ester tosilate.Obtaining 504mg (55%) title compound, is colorless solid.Mp89-91℃。 ESI-MS(m/e)460[M+H] +。IR(KBr)：3735，3649，3547，3377，2945，1730，1674，1522，1492，1465，1336，1266，1194，1158，1119，974，768，725，677。 1HNMR(300MHz，CDCl 3)8.9(d，J＝8.7Hz，2H)，8.53(d，J＝9.0Hz，1H)，8.22(d，J＝7.8Hz，1H)，7.65(d，J＝5.4Hz，2H)，7.41(m，6H)，5.78(s，2H)，?5.24(m，2H)，4.94(m，1H)，3.30(s，3H)，2.15(m，1H)，1.86(m，3H)，1.00(m，6H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝172.93，165.10，141.71，140.69，138.04，135.65，130.70，129.86，129.14，128.55，128.26，128.16，122.24，122.00，121.45，114.53，110.24，74.59，66.93，56.42，51.06，33.97，25.00，22.96，21.96。
Embodiment 13 prepares 9-methoxyl methyl-β-carboline-3-formyl Methionin benzyl ester (5i)
According to the method for embodiment 5, obtaining 290mg (31%) title compound from 979mg (2.4mmol) 1B benzyl ester tosilate, is colorless solid.Mp71-72℃。 ESI-MS(m/e)475[M+H] +。IR(KBr)：3648，3379，1868，1732，1647，1558，1457，1428，1340，1241，1189，950，867，741，486。 1HNMR(300MHz，CDCl 3)8.92(d，J＝18.5Hz，1H)，8.53(d，J＝27.9Hz，1H)，8.22(d，J＝7.8Hz，1H)，8.10(s，1H)，7.65(m，2H)，7.50(m，1H)，7.40(m，2H)，7.34(s，3H)，5.83(d，J＝8.1Hz，2H)，5.10(m，2H)，3.69(d，J＝8.7Hz，1H)，3.10(m，2H)，1.64(s，2H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝171.48，170.02，141.65，137.04，136.46，135.54，130.53，129.90，129.24，128.37，127.99，127.57，122.25，121.79，121.38，110.89，110.23，74.59，66.78，57.13，56.50，51.92，40.42，24.11，23.07。
Embodiment 14 prepares 9-methoxyl methyl-β-carboline-3-formylmethionine benzyl ester (5j)
According to the method for embodiment 5, obtaining 270mg (28%) title compound from 986mg (2.4mmol) METHIONINE benzyl ester tosilate, is light yellow solid.Mp98-99℃。 ESI-MS(m/e)478[M+H] +。IR(KBr)：3902，3755，3679，3648，3629，3587，3566，3375，2970，2941，2889，2820，1725，1682，1516，1457，1375，1338，1175，1119，1051，753，699，419。 1HNMR(300MHz，CDCl 3)8.92(d，J＝7.2Hz，2H)，8.73(d，J＝2.7Hz，1H)，8.22(d，J＝5.4Hz，1H)，7.65(s，2H)，7.41(m，6H)，5.80(s，2H)，5.25(m，2H)，5.04(m，1H)，3.30(s，3H)，2.51(m，2H)，2.31(m，1H)，2.21(m，1H)，2.09(s，3H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝172.93，165.10，141.71，140.69，138.04，135.65，130.70，129.86，129.14，128.55，128.26，128.16，122.24，122.00，121.45，114.53，110.24，74.59，66.93，56.42，51.06，33.97，25.00，22.96，21.96。
Embodiment 15 prepares 9-methoxyl methyl-β-carboline-3-formyl phenylalanine benzyl ester (5k)
According to the method for embodiment 5, obtaining 313mg (85%) title compound from 384mg (0.9mmol) L-Phe benzyl ester tosilate, is clear yellow viscous solid.Mp79-80℃。 ESI-MS(m/e)494[M+H] +。IR(KBr)：3948，3852，3749，3710，3648，3545，3502，2881，2821，2358，1733，1683，1657，1558，1507，1457，452。 1HNMR(300MHz，CDCl 3)8.92(s，2H)，8.63(d，J＝8.4Hz，1H)，8.22(d，J＝7.8Hz，1H)，7.68(s，2H)，7.28(m，10H)，5.80(s，2H)，5.20(m，3H)，5.04(m，1H)，3.30(m，5H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝171.50，164.93，141.70，140.61，138.06，136.12，135.35，130.78，129.81，129.41，129.13，128.54，12.51，128.46，128.35，126.94，122.25，122.02，121.45，114.44，110.23，74.61，67.08，56.45，53.57，49.11。
Embodiment 16 prepares 9-methoxyl methyl-β-carboline-3-formyl Serine benzyl ester (51)
According to the method for embodiment 5, obtaining 305mg (70%) title compound from 440mg (1.2mmol) Serine benzyl ester tosilate, is colorless solid.Mp92-93℃。 ESI-MS(m/e)434[M+H] +。IR(KBr)：3853，3751，3675，3629，3372，2940，2883，2821，1742，1640，1622，1532，1495，1463，1395，1263，1228，1201，1118，1062，756。 1HNMR(300MHz，CDCl 3)9.00(d，J＝7.5Hz，1H)，8.86(s，2H)，8.18(d，J＝7.8Hz，1H)，7.65(m，2H)，7.40(m，6H)，5.72(s，2H)，5.30(s，2H)，5.01(m，1H)，4.18(d，J＝3.0Hz，2H)3.30(s，3H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝170.36，165.74，141.63，140.23，138.00，135.32，130.72，129.73，129.21，128.64，128.44，128.17，122.24，121.87，121.50，114.59，110.26，74.52，67.47，63.97，56.46，55.43。
Embodiment 17 prepares 9-methoxyl methyl-β-carboline-3-formyl threonine benzyl ester (5m)
According to the method for embodiment 5, obtaining 314mg (29.3%) target compound from 914mg (2.4mmol) L-threonine benzyl ester tosilate, is white solid.Mp98-99℃。 ESI-MS(m/e)448[M+H] +。IR(KBr)：3948，3752，3745，3710，3676，3648，3545，3290，1748，1652，16222，1532，1496，1458，1399，1267，1254，1105，890，878。 1HNMR(300MHz，CDCl 3)8.92(d，J＝2.7Hz，2H)，8.86(m，1H)，8.18(d，J＝4.5Hz，1H)，7.66(m，2H)，7.38(m，6H)，5.79(s，2H)，5.28(d，J＝9.0Hz，2H)，4.95(m，1H)，4.53(s，1H)，3.32(s，3H)，2.40(m，1H)，1.36(d，J＝6.3Hz，3H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝170.95，165.9l，141.70，138.10，130.78，129.19，128.62，128.39，128.18，122.27，121.97，121.50，114.87，111.89，110.29，77.45，77.02，74.59，68.64，58.06。
Embodiment 18 prepares 9-methoxyl methyl-β-according to the method for embodiment 5, from carboline-3-formyl tryptophan benzyl ester (5n)
1120mg (2.4mmol) L-Trp benzyl ester tosilate obtains 310mg (29%) title compound, is light reddish brown color solid.Mp82-83℃。 ESI-MS(m/e)533[M+H] +。IR(KBr)：3931，3853，3801，3735，3649，3291，1733，1717，1653，1622，1521，1457，1338，1248，1191，1113，1052，744。 1HNMR(300MHz，CDCl 3)9.12(s，1H)，8.86(s，1H)，8.45(d，J＝7.8Hz，1H)，7.85(d，J＝8.4Hz，1H)，7.680(m，1H)，7.54(d，J＝7.8Hz，1H)，7.36(m，7H)，7.16(s，1H)，7.07(t，J＝14.7Hz，1H)，6.96(t，J＝14.7Hz，1H)，5.96(s，1H)，5.15(t，J＝15.3Hz，2H)，4.96(m，1H)，3.21(s，3H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝170.95，165.91，141.70，140.39，138.10，135.42，130.78，129.84，129.19，128.62，128.39，128.18，122.27，121.97，121.50，114.69，110.29，74.59，68.64，67.29，57.84，56.46，20.09。
Embodiment 19 prepares 9-methoxyl methyl-β-carboline-3-formyltyrosine benzyl ester (5o)
According to the method for embodiment 5, obtaining 280mg (55%) title compound from 531mgL-tyrosine benzyl ester tosilate, is colorless solid.Mp94-95℃。 ESI-MS(m/e)510[M+H] +。IR(KBr)：3940，3832，3745，3719，3608，3595，3512，2841，1757，1646，1533，1461，1167，1111，1058，768，733。 1HNMR(300MHz，CDCl 3)8.82(s，2H)，8.70(d，J＝8.1Hz，1H)，8.19(d，J＝7.8Hz，1H)，7.64(s，2H)，7.35(m，7H)，7.01(d，J＝7.8Hz，2H)，6.70(d，J＝7.8Hz，2H)，5.77(s，1H)，5.23(m，2H)，3.32(s，3H)，3.22(m，2H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝171.63，164.98，155.04，141.71，138.01，135.30，130.67，130.51，129.89，129.24，128.57，128.51，128.39，127.71，122.31，121.92，121.51，115.52，114.63，110.24，74.58，67.15，56.48，53.84，37.65。
Embodiment 20 prepares 9-methoxyl methyl-β-carboline-3-formyl α-amino-isovaleric acid benzyl ester (5p)
According to the method for embodiment 5, obtaining 370mg (42%) title compound from 910mg (2.4mmol) Valine benzyl ester tosilate, is thick pale yellow solid.Mp87-88℃。 ESI-MS(m/e)446[M+H] +。IR(KBr)：3853，3750，3734，3675，3648，3628，3587，3586，3545，3526，3502，2963，1733，1717，1669，1653，1558，1521，1507，1496，1471，1457，1361，1338，1248，1192，1152，1115，750，418。 1HNMR(300MHz，CDCl 3)8.92(d，J＝8.7Hz，2H)，8.68(d，J＝9.3Hz，1H)，8.23(d，J＝7.8Hz，1H)，7.65(s，2H)，7.35(m，6H)，5.80(s，2H)，5.25(m，2H)，4.90(m，1H)，3.63(m，1H)，3.36(s，3H)，0.97(m，6H)。 13C-NMR(DMSO-d 6，75MHz)：δ/ppm＝?171.94，165.21，141.72，140.70，138.05，135.60，130.69，129.89，129.16，128.56，128.33，122.28，122.01，121.46，114.56，110.25，74.61，66.90，57.42，56.44，31.67，19.24，17.86。
Experimental example 1 evaluates 5a-p anti-tumour cell proliferative activity
1640 substratum of 5a-p containing 0.25%DMSO are mixed with desired concn.K562, HL60, Hela, HepG2 and HT-29 five strain tumour cell equal purchased from American standard type culture collection institute (ATCC).RPMI-1640 culture medium dry powder, purchased from Gibco company, contains 8.2g NaCl, 0.2g KCl, 1.56g Na in often liter of PBS buffered soln 2hPO 4h 2o, 0.2gKH 2pO 4, pH value 7.4, foetal calf serum is purchased from Hyclone company, and 0.25% trypsin solution is purchased from Hyclone company, and penicillin and Streptomycin sulphate are purchased from solarbio company.Four tetrazolium bromides (MTT), purchased from solarbio company, are dissolved in PBS solution, make the solution of 5mg/mL, use after filtration sterilization, keep in Dark Place.Zorubicin (ADR) is purchased from Beijing Hua Feng United Technologies Corp., and DMSO (DMSO) is purchased from Hyclone company.HepG2 cell selects DMEM substratum, and other cells select RPMI-1640 substratum.All containing the foetal calf serum and 1 × 10 of 10% fire extinguishing in nutrient solution 5u/L penicillin and 100mg/L Streptomycin sulphate.
1) growth conditions is good, be in the attached cell A549 of logarithmic phase, HepG2, HT-29 are by 5 × 10 4sub 96 orifice plates of cell density inoculation of individual/mL, every hole 100 μ L.Be placed in 37 DEG C, 5%CO 2cultivating in incubator treats adherent in 4 hours.Every hole adds the 5a-p solution of 25 μ L through sterilising treatment by the concentration gradient preset, and control wells adds 25 μ L RPMI-1640, parallel 6 holes.At 37 DEG C, 5%CO 2cultivate 48 hours in incubator, every hole adds the MTT solution that 25 μ L concentration are 5mg/mL, continues to be placed in 37 DEG C, 5%CO 2cultivate 4 hours in incubator.Careful sucking-off supernatant liquor, every hole adds 100 μ LDMSO dissolve purple residues (first a ceremonial jade-ladle, used in libation), and Oscillating Flat makes precipitation all dissolve in 10 minutes, measures OD value (absorbancy), wavelength 570nm in 570nm microplate reader.
According to relative survival rate=(5a-p OD value-RPMI-1640OD value)/(it is blank that D contrasts-D) × 100% " calculate the inhibiting rate of the sample under each sample concentration to tumour cell.
Test parallel repetition 3 times, with inhibiting rate, compound concentration is mapped, calculate the IC of 5a-p 50value.
2) growth conditions is good, be in suspension cell K562 and HL60 of logarithmic phase by 5 × 10 4the cell density of individual/mL is inoculated in 96 orifice plates, every hole 100 μ L.Be placed in 37 DEG C, 5%CO 2cultivating in incubator treats adherent in 4 hours.Every hole by preset concentration gradient add the 5a-p solution of 25 μ L through sterilising treatment, control wells adds 25 μ L RPMI-1640, parallel 6 holes at 37 DEG C, 5%CO 2cultivate 48 hours in incubator, every hole adds the MTT solution that 25 μ L concentration are 5mg/mL, continues to be placed in 37 DEG C, 5%CO 2cultivate 4 hours in incubator.Careful sucking-off supernatant liquor, every hole adds 100 μ LDMSO dissolve purple residues (first a ceremonial jade-ladle, used in libation), and Oscillating Flat makes precipitation all dissolve in 10 minutes, measures OD value (absorbancy), wavelength 570nm in 570nm microplate reader.
According to relative survival rate=(5a-p OD value-RPMI-1640OD value)/(it is blank that D contrasts-D) × 100% " calculate the inhibiting rate of the sample under each sample concentration to tumour cell.
Test parallel repetition 3 times, with inhibiting rate, compound concentration is mapped, calculate the IC of 5a-p 50value.
Experimental result lists table 1 in.Data declaration 5a-p to K562, HL60, Hela, HepG2 and HT-29 five strain tumour cell all only show faint cytotoxicity, embody creativeness of the present invention.
Activity (the IC of table 15a-p inhibition tumor cell propagation 50, μM)
Experimental example 2 evaluates the activity of 5a-p Tumor suppression growth
5a-p adds a small amount of tween 80 hydrotropy, adds physiological saline gradually to desired concn, and single intraperitoneal injection dosage is 0.1 μm of ol/kg.Zorubicin is normal saline solution, and single intraperitoneal injection dosage is 1 μm of ol/kg..Zorubicin is normal saline solution.5a-p and Zorubicin all successive administrations 7 days, altogether administration 7 times.
Inoculation eugonic S180 ascitic tumor knurl liquid after 7 days is extracted under aseptic condition, the liquid of (1: 3) is become fully to mix with normal saline dilution, by freshly prepared 0.2% Trypan Blue of tumor cell suspension, by white blood cell count(WBC) method counting after mixing, contaminate blue person for dead cell, tinter is not viable cell, and is calculated as follows cell concn and cell survival rate.Adopt anti-tumor in vivo armpit subcutaneous vaccination model.Knurl liquid homogenate method survival rate being greater than 90% is prepared into 1.5 × 10 7the cell suspension of individual/mL, in mouse armpit subcutaneous vaccination 0.2mL/ only, causes solid tumor animal model.
Inoculation eugonic S180 ascitic tumor knurl liquid after 7 days is extracted under aseptic condition, the liquid of (1: 3) is become fully to mix with normal saline dilution, by freshly prepared 0.2% Trypan Blue of tumor cell suspension, by white blood cell count(WBC) method counting after mixing, contaminate blue person for dead cell, tinter is not viable cell, and presses cell concn=(in 4 block plaid viable count/4) × 10 4extension rate=cell count/mL and cell survival rate=viable count/(viable count+dead cell number) × 100% calculate cell concn and cell survival rate.
ICR male mice (cleaning grade, often organize 12 mouse, body weight is 20 ± 2g).Knurl liquid homogenate method S180 viable cell survival rate being greater than 90% is prepared into 1.5 × 10 7the cell suspension of individual/mL, in the subcutaneous vaccination of ICR male mice armpit (0.2mL/ only), causes lotus S180 solid tumor mouse and accepts 5a-p and doxorubicin.Blank is the physiological saline (dosage is 3ml/kg) containing a small amount of tween 80.Every day abdominal injection 1 time, successive administration 7 days.Treatment, to the 8th day, claims Mouse Weight, and de-cervical vertebra puts to death mouse, and takes the tumour of each group of mouse and each main organs is weighed, and finally adds up the knurl body when dirty body ratio of each group of medicine.Knurl body ratio=knurl heavy (mg)/put to death front body weight (g).This experimental data statistics all adopts t inspection and variance analysis.Experimental result table 2 represents.
Data show that 5a-p can suppress mice bearing S180 tumor growth effectively under 1 μm of ol/kg dosage, embodies creativeness of the present invention.
Table 25a-p is on the impact of mice bearing S180 tumor weight a
Note: a) n=12; A) with physiological saline group than P < 0.01, with Zorubicin than p > 0.05; B) with physiological saline group than P < 0.01.

Claims (4)

1. the 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester of formula 1, the AA in formula I is selected from L-Leu, L-Glu, L-Val, L-Trp, L-Arg, L-Tyr, L-Met, L-Phe, L-Ala, L-Gly, L-His, L-Asp, L-Ile, L-Lys, L-Ser and L-Thr residue.
Disclose them.Disclose them to HL60 (human promyelocytic leukemia), K562 (human chronic polymorpho nuclear leukemia cells), the restraining effect that HeLa (cervical cancer cell of epithelial origin), HepG2 (hepatocellular carcinoma cells) and HT-29 (human colon cancer cell) breed.Further disclose their restraining effect to mice bearing S180 tumor growth.Thus the invention discloses the potential applicability in clinical practice of 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester as antitumor drug of formula I representative.
2. the preparation method of the 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester of claim 1, the method comprises:
1) L-Trp is changed into 3S-9-methylol-tetrahydro-beta-carboline-3-carboxylic acid (1);
2) 3S-9-methylol-tetrahydro-beta-carboline-3-carboxylic acid (1) is changed into 3S-9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylate methyl ester (2);
3) make to change 9-methoxyl methyl-β-carboline-3-carboxylate methyl ester (3) into potassium permanganate oxidation 3S-9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylate methyl ester (2);
4) 9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylate methyl ester (3) is changed into 9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylic acid (4) in the basic conditions;
5) 9-methoxyl methyl-tetrahydro-beta-carboline-3-carboxylic acid (4) and 16 seed amino acid benzyl esters are obtained by reacting 16 kinds of 9-methoxyl methyls-β-carboline-3-formyl amino acid formyl amino acid benzyl ester (5a-p).
3., according to method described in claim 2, it is characterized in that step 5) described in amino-acid benzyl ester be selected from L-Leu-OBzl, L-Glu-OBzl, L-Val-OBzl, L-Trp-OBzl, L-Arg-OBzl, L-Tyr-OBzl, L-Met-OBzl, L-Phe-OBzl, L-Ala-OBzl, Gly-OBzl, L-His-OBzl, L-Asp (OBzl)-OBzl, L-Ile-OBzl, L-Lys-OBzl, L-Ser-OBzl and L-Thr-OBzl.
4. the compound of claim 1 and 2 is as the application of antineoplastic agent.
CN201310225711.4A 2013-06-05 2013-06-05 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, antitumor activity and application thereof Pending CN104231042A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201310225711.4A CN104231042A (en) 2013-06-05 2013-06-05 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, antitumor activity and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201310225711.4A CN104231042A (en) 2013-06-05 2013-06-05 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, antitumor activity and application thereof

Publications (1)

Publication Number Publication Date
CN104231042A true CN104231042A (en) 2014-12-24

Family

ID=52220030

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201310225711.4A Pending CN104231042A (en) 2013-06-05 2013-06-05 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, antitumor activity and application thereof

Country Status (1)

Country Link
CN (1) CN104231042A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107629056A (en) * 2015-06-23 2018-01-26 首都医科大学 Benzimidazole quinazoline Salicyloy lamino acid benzyl ester, its preparation, antitumor activity and application

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101591335A (en) * 2008-05-30 2009-12-02 首都医科大学 N-(L-aminoacyl)-1,2,3,4-tetrahydrochysene carboline acyl aminoacid benzyl ester and synthetic method thereof and application
CN101906102A (en) * 2009-06-02 2010-12-08 首都医科大学 Beta-carboline alkaloid derivative, preparation method and application thereof
CN102250202A (en) * 2010-05-19 2011-11-23 首都医科大学 1-paranitrophenyl-beta-carboline-3-formyl amine acid carbamates and synthesis method and use thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101591335A (en) * 2008-05-30 2009-12-02 首都医科大学 N-(L-aminoacyl)-1,2,3,4-tetrahydrochysene carboline acyl aminoacid benzyl ester and synthetic method thereof and application
CN101906102A (en) * 2009-06-02 2010-12-08 首都医科大学 Beta-carboline alkaloid derivative, preparation method and application thereof
CN102250202A (en) * 2010-05-19 2011-11-23 首都医科大学 1-paranitrophenyl-beta-carboline-3-formyl amine acid carbamates and synthesis method and use thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
RIHUI CAO: "Synthesis, acute toxicities, and antitumor effects of novel 9-substituted β-carboline derivatives", 《BIOORGANIC & MEDICINAL CHEMISTRY》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107629056A (en) * 2015-06-23 2018-01-26 首都医科大学 Benzimidazole quinazoline Salicyloy lamino acid benzyl ester, its preparation, antitumor activity and application
CN107629056B (en) * 2015-06-23 2020-01-14 首都医科大学 Benzimidazole quinazoline salicyloyl amino acid benzyl ester, preparation thereof, anti-tumor activity thereof and application thereof

Similar Documents

Publication Publication Date Title
CN103450329B (en) 3H-imidazo[4,5-c]pyridine-6-formyl-amido acid benzyl esters and their synthesis, anti-tumor activity and use
CN103159828B (en) 1-(4-hydroxy-3-methoxycarbonyl)-beta-carboline-3-formyl tryptophyl amino acid benzyl ester, and synthesis and application thereof
CN103450199B (en) Theanine modify carboline acyl aminoacid benzyl ester, its preparation, anti-tumor activity and application
JP6929234B2 (en) Pharmaceutical co-crystal composition and its uses
CN105198961A (en) RGD modified 5-fluorouracil and preparation method, nanostructure, activity and application thereof
CN105218634A (en) The indoles quinolizine that YIGSR modifies, its preparation, nanostructure, active and application
CN105218637A (en) The indoles quinolizine that LPNISKP modifies, its preparation, nanostructure, active and application
CN111848544A (en) Amino acid derivative capable of being traced by fluorescence and preparation method and application thereof
Kamaludin et al. Cytotoxicity evaluation and the mode of cell death of K562 cells induced by organotin (IV)(2-methoxyethyl) methyldithiocarbamate compounds
CN104231042A (en) 9-methoxymethyl-beta-carboline-3-formyl amino acid benzyl esters, and synthesis, antitumor activity and application thereof
CN104211700B (en) Carboline carboxylate analog, its synthesis, nanostructured, anti-tumor activity and application
CN106083925B (en) Diphosphonic acid compound and preparation method and application thereof
CN103509011B (en) 1-(4-hydroxyl-3-formamido group acidic group-phenyl)-β-carboline, preparation, anti-inflammatory and anti-tumor activity and application
CN110041342A (en) A kind of selenium-containing compound and application thereof
Singh et al. Cancer stem cell activity of copper (ii)-terpyridine complexes with aryl sulfonamide groups
CN108030777B (en) Chloroguanide application in preparation of anti-tumor drugs
Varol et al. Anti-lung cancer and anti-angiogenic activities of new designed boronated phenylalanine metal complexes
CN105294835A (en) LDV modified 5-fluorouracil, preparation therefor, nanostructure thereof, vigor thereof and application thereof
CN110003142A (en) A kind of synthesis and application thereof
WO2016070712A1 (en) Amino acid derivatives and applications thereof
CN105237618A (en) Amino acid benzyl ester-modified beta-carbolines, synthesis, nanostructures, activities, and applications
CN102250190A (en) Oleanolic saponin derivative, preparation method of salt thereof and application of derivative
CN103450198B (en) Imidazo pyrido imidazole-3-substituted benzyl acetates, and synthesis, antineoplastic activities and application thereof
US20200361973A1 (en) Combination product comprising dicycloplatin and preparation method and use thereof
CN106146615A (en) Imidazopyridine-6-formyl-amino-acid benzyl ester, its synthesis, activity and application

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20141224