CN102250203B - Beta-carboline aminoacid benzyl ester, preparation method and application thereof - Google Patents
Beta-carboline aminoacid benzyl ester, preparation method and application thereof Download PDFInfo
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- CN102250203B CN102250203B CN201010177167.7A CN201010177167A CN102250203B CN 102250203 B CN102250203 B CN 102250203B CN 201010177167 A CN201010177167 A CN 201010177167A CN 102250203 B CN102250203 B CN 102250203B
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Abstract
The invention relates to a beta-carboline aminoacid benzyl ester, a preparation method and application thereof. According to the invention, L-tryptophan and formaldehyde are treated with Pictet-Spengler condensation under catalysis of dilute sulphuric acid to obtain a series of intermediates; and the intermediates are coupled with a plurality of aminoacid benzyl esters to obtain 2-[tetrahydro-beta-carbolino(2,2-dimethyl-imidazole-4-ketone-3-base)]-4-(methyl propyl)- valeryl aminoacid benzyl ester. The method of the invention is simply operated; raw materials are easily available; and the obtained derivative has excellent antitumor activity in vitro and in vivo. Therefore, the invention discloses 18 varieties of novel 2-[tetrahydro-beta-carbolino(2,2-dimethyl-imidazole-4-ketone-3-base)]-4-(methyl propyl)- valeryl aminoacid benzyl esters to be used as future clinic antitumor drugs.
Description
Technical field
The present invention relates to a series antineoplastic medicament, be particularly related to β-carboline amino acid benzyl ester, be 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeryl amino-acid benzyl ester and its preparation method and application, biomedicine field belonged to.
Background technology
Malignant tumour is that a kind of common disease and frequently-occurring disease ，Jin 20Nian Lai， China tumor mortality rate of serious threat human health risen 29.42%.In the middle prime of life crowd of 35 to 59 years old, tumour has been listed as and has occupied first of all kinds of causes of the death.Data show: China's tumor incidence is about 2,00/,100,000 people, more than annual new cases approximately 2,200,000 people, more than controlling patient approximately 6,000,000 people.The methods for the treatment of of tumour has operative treatment, radiotherapy and pharmacological agent (chemotherapy).At present, chemotherapy remains the Main Means of clinical treatment tumour.Seeking antitumor medicine is one of focus of new drug research.β-carboline analog derivative has biologic activity widely, and they can suppress the synthetic of topoisomerase, cell cycle protein dependent kinase (CDK) and DNA, and in energy intercalation of DNA two strands, all these effects make them have very strong anti-tumor activity.Known, 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid has certain anti-tumor activity, therefore, at 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)] introduce natural amino acid benzyl ester on the carboxyl of-4-methyl-valeric acid and likely produce more outstanding anti-tumor activity.Based on above theory, the invention discloses and 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)] invention that-4-methyl-valeryl amino-acid benzyl ester is relevant.
Summary of the invention
The object of the present invention is to provide a 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeryl amino-acid benzyl ester, has confirmed the application of these derivatives in preparing antitumor drug by the inside and outside antitumor activity to them.
The present invention is achieved through the following technical solutions this purpose:
The compound of structural formula 3,4,5 and 6a-r representative,
Wherein R is CH
3, H, CH
2cH (CH
3)
2, CH (CH
3) CH
2cH
3, CH (CH
3)
2, CH
2c
6h
5, CH
2cH
2cO
2bzl, CH
2cO
2bzl, CH
2oH, CHOHCH
3, (CH
2)
4nH
2, (CH
2)
2cONH
2, CH
2cONH
2, (CH
2)
2sCH
3, CH
2sC (CH
3)
3, Indole-5-yl-CH
2, CH
2-imidazole or CH
2-C
6h
4-OH-p.
The present invention also provides a kind of 3S-N-Boc-1 of preparation, 2,3, the method of 4-tetrahydro-beta-carboline-3-formyl-L-Leu benzyl ester (3), be 3S-N-Boc-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid and TosHLeu-OBzl, under the existence of I-hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (DCC) and N-methylmorpholine (NMM), react under condition of ice bath.
The present invention also provides a kind of 2-[of preparation tetrahydro-beta-carboline also (2, the method of 2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-benzyl valerianate (4), be 3S-N-Boc-1,2,3,4-tetrahydro-beta-carboline-3-formyl-L-Leu benzyl ester (3), under hydrochloric acid and ethyl acetate existence, takes off Boc under condition of ice bath.
The present invention also provides a kind of 2-[of preparation tetrahydro-beta-carboline also (2, the method of 2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), be 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-benzyl valerianate (4) hydrogenolysis debenzylation under palladium/carbon exists.
The invention provides a kind of 2-[of preparation tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)] method of-4-methyl-valeryl amino-acid benzyl ester (6a-r), it is 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), under the existence of DCC, HOBt and NMM, reacts with various amino-acid benzyl esters under condition of ice bath.
The present invention also provides 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-benzyl valerianate (4) is as preparing the application of antitumor drug.
The present invention also provides 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeryl amino-acid benzyl ester (6a-r) is as preparing the application of antitumor drug.
The synthetic route of the inventive method as shown in Figure 1.
In Fig. 1, i) dilute sulphuric acid, formaldehyde; Ii) (Boc)
2o, DMF, Et
3n; Iii) DCC, HOBt, NMM, anhydrous tetrahydro furan (THF); Iv) 4N HCl/EtOAc, methyl alcohol, acetone, Et
3n; v)H
2, Pd/C, methyl alcohol; Vi) DCC, HOBt, NMM, anhydrous THF, amino-acid benzyl ester.R=CH in 6a
3; R=H in 6b; R=CH in 6c
2cH (CH
3)
2; R=CH (CH in 6d
3) CH
2cH
3; R=CH (CH in 6e
3)
2; R=CH in 6f
2c
6h
5; R=CH in 6g
2cH
2cO
2bzl; R=CH in 6h
2cO
2bzl; R=CH in 6i
2oH; R=CHOHCH in 6j
3; R=(CH in 6k
2)
4nH
2; R=(CH in 6l
2)
2cONH
2; R=CH in 6m
2cONH
2; R=(CH in 6n
2)
2sCH
3; R=CH in 6o
2sC (CH
3)
3; R=Indole-5-yl-CH in 6p
2; R=CH in 6q
2-imidazole; R=CH in 6r
2-C
6h
4-OH-p.
The present invention obtains a series of intermediates by L-Trp is carried out to Pictet-Spengler condensation with formaldehyde under dilute sulphuric acid catalysis, again intermediate and multiple amino acids benzyl ester are carried out to coupling, obtain 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeryl amino-acid benzyl ester derivative.The inventive method is simple to operate, and raw material is easy to get, and gained derivative has outstanding inside and outside anti-tumor activity, is conducive to the development of antitumor drug research and development.
Accompanying drawing explanation
Fig. 1 is the synthetic route of the inventive method.
Embodiment
Below in conjunction with embodiment, the invention will be further described, it should be understood that these embodiment, only for the object of illustration, never limit the scope of the invention.
Embodiment 1 preparation 3S-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (1)
200mL water is placed in to the round-bottomed flask of 250mL, slowly adds the 0.1mL vitriol oil.In the dilution heat of sulfuric acid obtaining, add 2.520g (12.4mmol) L-Trp sonic oscillation to L-Trp to dissolve completely.Toward adding 5mL concentration in the solution obtaining, it is the formalin of 37-40%.Reaction mixture stirring at room 6 hours, thin-layer chromatography monitors L-Trp and disappears, termination reaction.In reaction soln, slowly drip strong aqua, regulate the pH to 6 of reaction mixture, standing half an hour.Büchner funnel decompression leaches the throw out of generation, water and acetone is drip washing gained throw out repeatedly, and drip washing 3 times, is laid in culture dish by the colorless solid leaching, and be placed in after stink cupboard dries up and obtain 2.640g (98.9%) title compound, be colourless powder.ESI-MS(m/e)215[M-H]
-。
Embodiment 2 preparation 3S-N-Boc-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (2)
In 100mL eggplant bottle, add 20mL DMF, add 2.033g (9.41mmol) 3S-1 under stirring, 2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (1), obtains white opacity suspension.Ice bath adds 2.752g (12.6mmol) Boc under stirring in this suspension
2o, adds triethylamine reacting liquid pH value is adjusted to 10.Reaction mixture stirring at room 48 hours, thin-layer chromatography is monitored to the disappearance of S-carboline carboxylate, termination reaction.Pour reaction solution into watch-glass, in fan blowing down, within approximately 24 hours, extremely do.By the oily matter drying up 100mL acetic acid ethyl dissolution, be then placed in 250mL separating funnel, use KHSO
4(5%) solution washing (10mL * 3).Separate the ethyl acetate layer of merging, in 150mL triangular flask, add anhydrous sodium sulfate drying 2 hours, filtration under diminished pressure.Filtrate decompression is concentrated into dry, separates out white solid.In the white solid obtaining, add chloroform, filtration under diminished pressure, obtains 2.230g (75.0%) title compound, is colourless powder.ESI-MS(m/e)315[M-H]
-。
Embodiment 3 preparation 3S-N-Boc-1,2,3,4-tetrahydro-beta-carboline-3-formyl-L-Leu benzyl ester (3)
Ice bath is cooling adds 2.243g (7.10mmol) 3S-N-Boc-1,2,3 down in 250mL eggplant bottle, 4-tetrahydro-beta-carboline-3-carboxylic acid (2), 4.900g (12.4mmol) TosHLeu-OBzl, 1.402g (10.4mmol) HOBt, add the anhydrous THF of 80mL, stirring and dissolving.2.589g (12.6mmol) DCC is dissolved in the anhydrous THF of 10mL, slowly drops in reaction flask under ice bath cooling conditions, fully stirs 5 minutes.NMM regulates reaction solution pH to 8, and reaction mixture stirs 2 filtering dicyclohexylurea (DCU) of suction filtration (DCU) after 4 hours at 0 ℃, and filtrate decompression is concentrated into dry.By 150mL acetic acid ethyl dissolution for the resistates obtaining, be then placed in 250ml separating funnel, with saturated sodium bicarbonate aqueous solution, wash (30mL * 3), saturated sodium-chloride water solution successively and wash (30mL * 3), concentration is 5% by weight aqueous potassium hydrogen sulfate and wash (30mL * 3), saturated sodium-chloride water solution and wash that (30mL * 3), saturated sodium bicarbonate aqueous solution are washed (30mL * 3), saturated sodium-chloride water solution is washed (30mL * 3).Combined ethyl acetate layer with anhydrous sodium sulfate drying 2 hours, then filters, filtrate decompression is concentrated into dryly, obtains faint yellow oily matter.Add a small amount of ether dissolution, water pump decompressing and extracting is solidified, 3 times repeatedly.Obtaining 3.565g (96.7%) title compound, is colourless powder.R
f=0.65 (chloroform: methyl alcohol=20:1), ESI-MS (m/e) 520[M+H]
+.
1h-NMR (DMSO-d
6, 300MHz): δ=10.822 (d, J=18.9Hz, 1H), 8.397 (m, 1H); 7.293 (s, 5H), 7.208 (m, 2H), 6.991 (m, 2H); 5.104 (m, 1H), 4.941 (m, 2H), 4.694 (d, J=18.9Hz; 1H), 4.542 (m, 1H), 3.230 (m, 1H); 3.040 (m, 1H), 1.570 (m, 3H), 1.423 (s; 9H), 0.839 (s, 3H), 0.764 (s, 3H);
13c-NMR (DMSO-d
6, 75MHz): δ=172.56,172.02,155.54,136.55,136.29,131.10,128.80,128.42,128.22,126.87,121.12,118.83,117.79,111.35,104.41,103.85,80.15,79.99,66.38,53.50,52.23,50.75,50.41,33.83,28.41,24.93,24.59,24.17,23.37,23.17,15.64.
Embodiment 4 preparation 2-[tetrahydro-beta-carbolines are (2,2-dimethyl-imidazol-4-one-3-yl) also]-4-methyl-benzyl valerianate (4)
By 2.147g (4.14mmol) 3S-N-Boc-1,2,3,4-tetrahydro-beta-carboline-3-formyl-L-Leu benzyl ester (3) is placed in 100mL eggplant-shape bottle, adds 5mL acetic acid ethyl dissolution.Under ice bath, in the solution obtaining, slowly splash into 20mL4N HCl/EtOAc.Reaction mixture stirs water pump after 2 hours and drains at 0 ℃.Residue adds ethyl acetate fully to stir, and water pump decompressing and extracting repeats 3 times.Add ether fully to stir rear water pump decompressing and extracting, repeat 3 times.The colourless powder obtaining is dissolved in the mixed solvent of 60mL methyl alcohol and 20mL acetone, with triethylamine, adjusts pH value to 9, lucifuge reaction 7 days, utilizes thin-layer chromatography to monitor to the disappearance of N-Boc-S-carboline acyl-L-Leu benzyl ester, termination reaction.By reaction mixture be evaporated to dry, for residue, 200mL acetic acid ethyl dissolution, solution are placed in 250mL separating funnel wash (30mL * 8) with 30mL saturated sodium-chloride water solution.Ethyl acetate layer after washing filters after 2 hours with anhydrous sodium sulfate drying.Filtrate decompression is concentrated into dry.For residue, 5mL dissolve with methanol, standing, then leaches 0.35g (37.7%) title compound, and it is pale yellow powder.R
f=0.32 (sherwood oil: acetone, 4:1), ESI-MS (m/e) 460[M+H]
+; Mp190-192 ℃;
1h-NMR (CDCl
3), 300MHz δ=7.882 (s, 1H), 7.524 (d, J=8.1Hz, 1H), 7.341 (m, 6H), 7,154 (m, 2H), 5.204 (m, 2H), 3.962 (m, 2H), 3.846 (d, J=13.5Hz, 1H), 3.481 (m, 1H), 3.190 (m, 1H), 2.769 (m, 2H), 1.612 (m, 1H), 1.461 (s, 3H), 1.245 (s, 3H), 0.955 (m, 6H).
13c-NMR (CDCl
3, 75MHz) δ=171.78,170.62,136.29,135.33,131.02,128.74,128.66,128.51,128.27,127.26,121.75,119.68,118.22,110.68,108.47,78.92,66.87,61.12,57.42,41.60,34.07,29.69,25.65,25.03,23.89,18.87,16.59,11.22.
By 131mg (0.285mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-benzyl valerianate (4) is placed in 100mL eggplant bottle, add 15mL dissolve with methanol, under stirring, add 15mg Pd/C, connect good threeway and hydrogen gas bag, after water pump decompression deflates, pass into hydrogen, repeatedly repeat 2 times.Keep passing into the state stirring reaction 2 hours of hydrogen.Thin-layer chromatography monitoring is to 4 disappearances, stopped reaction, and careful filtering Pd/C, concentrates filtrate to dryly, obtains 102mg (96.9%) title compound, and it is pale yellow powder.R
f=0.40 (chloroform: methyl alcohol: Glacial acetic acid=20:1:0.10), ESI-MS (m/e) 368[M-H]-; Mp236-237 ℃;
(c=0.60, CH
3oH);
1h-NMR (DMSO-d
6, 300MHz) δ=10.979 (s, 1H), 7.356 (dd, J
1=30.3Hz, J
2=7.6Hz, 2H), 6.988 (m, 2H); 3.928 (m, 2H), 3.768 (d, J=14.1Hz; 1H), 3.416 (m, 1H), 2.870 (m; 1H), 1.998 (m, 2H), 1.657 (m; 2H), 1.420 (s, 3H), 1.287 (s; 3H), 0.930 (t, 6H);
13c-NMR (DMSO-d
6, 75MHz) δ=172.73,171.00,136.53,133.17,127.17,120.94,118.83,117.91,111.41,106.34,78.55,57.23,52.77,47.05,41.77,38.95,33.80,25.81,25.20,24.94,24.90,23.85,23.22,22.77,19.70.
Embodiment 6 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetylalanine benzyl esters (6a)
Ice bath is cooling lower to adding 176mg (0.48mmol) 2-[tetrahydro-beta-carboline also (2 in 50mL eggplant bottle, 2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 65mg (0.48mmol) HOBt, 207mg (0.59mmol) TosHAla-OBzl, adds the anhydrous THF of 25mL to dissolve and stirs.118mg (0.57mmol) DCC is dissolved in the anhydrous THF of 5mL, under ice bath is cooling, drops in reaction flask, fully stirs 5 minutes.It is 8 that NMM regulates reaction solution pH, and reaction mixture stirs after 4 hours and utilizes TLC to detect 5 disappearances at 0 ℃, and stopped reaction, by 2 filtering DCU of reaction solution suction filtration, concentrates filtrate to dry.Gained resistates 30mL acetic acid ethyl dissolution, and be transferred in 100mL separating funnel, with saturated sodium bicarbonate aqueous solution, wash (10mL * 3), saturated sodium-chloride water solution successively and wash (10mL * 3), 5% aqueous potassium hydrogen sulfate and wash (10mL * 3), saturated sodium-chloride water solution and wash that (10mL * 3), saturated sodium bicarbonate aqueous solution are washed (10mL * 3), saturated sodium-chloride water solution is washed (10mL * 3).Combined ethyl acetate layer with anhydrous sodium sulfate drying 2 hours, then filters, and concentrates filtrate to dryly, obtains faint yellow oily matter.In gained oily matter, add a small amount of sherwood oil repeatedly to wear away 3 times, obtain 235mg (93.0%) title compound, it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 140mg (60%).R
f=0.27 (sherwood oil: acetone=2:1).ESI-MS(m/e)531[M+H]
+;Mp128-129℃;
(c=0.65,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.868(s,1H),8.238(d,J=6.3Hz,1H),7.428(d,J=6.3Hz,1H),7.321(s,6H),7.005(m,2H),5.096(s,2H),4.374(m,1H),3.961(m,2H),3.742(d,J=14.1Hz,1H),3.454(d,J=8.1Hz,1H),2.917(d,J=14.7Hz,1H),2.071(m,1H),1.890(m,1H),1.701(m,1H),1.509(m,1H),1.362(s,3H),1.314(d,J=6.9Hz,3H),1.269(s,3H),0.910(s,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.49,172.24,171.08,136.40,136.24,132.85,128.83,128.47,128.31,127.15,121.04,118.92,117.98,111.36,106.38,79.29,66.53,57.52,55.87,48.16,47.87,41.72,39.03,33.77,25.33,24.97,24.14,23.08,22.74,18.43,17.14。
Embodiment 7 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl glycine benzyl esters (6b)
According to the operation of preparation 6a, by 359mg (0.97mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 403mg (1.19mmol) TosHGly-OBzl, 130mg (0.96mmol) HOBt, 245mg (1.19mmol) DCC prepares 462mg (92.0%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 280mg (60%).R
f=0.53 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)517[M+H]
+;Mp86-87℃;
(c=0.60,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.878(s,1H),8.147(m,1H),7.418(d,J=7.5Hz,1H),7.344(s,5H),7.322(m,1H),7.001(m,2H),5.110(s,2H),3.956(m,2H),3.932(m,1H),3.754(d,J=13.8Hz,1H),3.453(m,1H),2.906(m,1H),2.099(m,1H),1.934(m,1H),1.722(m,1H),1.562(m,2H),1.388(s,3H),1.274(s,3H),0.921(s,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.19,171.83,169.96,136.58,136.27,133.01,128.84,128.50,128.46,127.18,126.87,121.01,118.90,117.90,111.42,106.44,79.24,66.36,57.53,56.02,47.99,41.52,39.00,33.81,25.80,25.31,23.99,23.04,22.77,18.41。
Embodiment 8 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-N-Acetylleucine benzyl esters (6c)
According to the operation of preparation 6a, by 402mg (1.09mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 528mg (1.34mmol) TosHLeu-OBzl, 148mg (1.09mmol) HOBt, 275mg (1.33mmol) DCC prepares 570mg (91.5%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 350mg (62%).R
f=0.52 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)573[M+H]
+;Mp116-117℃;
(c=0.60,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.892(s,1H),8.222(d,J=7.8Hz,1H),7.430(d,J=7.5Hz,1H),7.325(m,1H),7.312(s,5H),7.004(m,2H),5.091(s,2H),4.367(m,1H),3.954(m,2H),3.750(d,J=13.8Hz,1H),3.458(m,1H),2.916(m,1H),2.100(m,1H),1.870(m,1H),1.725(m,1H),1.624(m,2H),1.611(m,1H),1.350(s,3H),1.266(s,3H),0.910(s,3H),0.903(s,6H),0.836(d,J=5.7Hz,3H).
13C-NMR(DMSO-d
6,75MHz)δ=172.40,172.29,171.53,136.54,136.20,133.01,128.84,128.50,128.40,127.15,121,02,118.90,117.97,111.42,106.38,79.26,66.55,57.53,56.14,50.83,47.97,41.77,39.00,33.82,25.79,25.29,24.95,24.73,24.12,23.27,23.08,22.73,21.51,18.15。
Embodiment 9 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl Isoleucine benzyl esters (6d)
According to the operation of preparation 6a, by 308mg (0.84mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 394mg(1.00mmol) TosHIle-OBzl, 132mg (0.98mmol) HOBt, 225mg (1.09mmol) DCC prepares 462mg (96.9%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 270mg (59%).R
f=0.51 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)573[M+H]
+;Mp96-97℃;
(c=0.65,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.886(s,1H),8.024(d,J=8.1Hz,1H),7.436(d,J=7.5Hz,1H),7.197(s,5H),7.307(m,1H),7.008(m,2H),5.086(m,2H),4.328(m,1H),3.922(m,2H),3.770(d,J=13.5Hz,1H),3.499(m,1H),2.932(m,1H),2.052(m,2H),1.932(m,1H),1.726(m,1H),1.556(m,3H),1.346(s,3H),1.290(s,3H),0.932(d,J=6.3Hz,6H),0.836(m,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.73,171.80,171.32,136.58,136.11,133.00,128.79,128.56,128.51,127.16,121,03,118.91,117.97,111.43,106.33,79.37,66.53,57.53,56.92,56.69,41.82,38.98,37.00,33.81,25.80,25.42,25.04,24.91,24.14,23.01,22.83,18.32,16.04,11.64。
Embodiment 10 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetylvaline benzyl esters (6e)
According to the operation of preparation 6a, by 602mg (1.63mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 745mg (1.96mmol) TosHVal-OBzl, 221mg (1.64mmol) HOBt, 415mg (2.01mmol) DCC prepares 846mg (93.1%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 367mg (43%).R
f=0.51 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)559[M+H]
+;Mp100-101℃;
(c=0.70,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.907(s,1H),8.013(d,J=8.4Hz,1H),7.438(d,J=7.5Hz,1H),7.301(s,5H),7.289(m,1H),7.006(m,2H),5.094(t,2H),4.278(m,1H),3.926(m,2H),3.766(d,J=13.8Hz,1H),3.506(m,1H),2.936(m,1H),2.121(m,2H),1.992(m,1H),1.720(d,J=11.7Hz,1H),1.543(m,1H),1.351(s,3H),1.292(s,3H),0.940(s,3H),0.920(s,3H),0.892(d,J=7.2Hz,3H),0.835(d,J=6.6Hz,3H).
13C-NMR(DMSO-d
6,75MHz)δ=172.73,171.88,171.35,136.54,136.13,133.00,128.81,128.58,128.53,127.13,121.04,118.91,118.01,111.42,106.32,79.37,66.55,57.51,57.45,56.88,41.82,33.82,30.50,25.41,25.79,25.02,24.93,24.16,23.05,22.82,19.44,18.33,17.85。
Embodiment 11 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetylphenylalanine benzyl esters (6f)
According to the operation of preparation 6a, by 792mg (2.15mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 1100mg (2.58mmol) TosHPhe-OBzl, 298mg (2.21mmol) HOBt and 539mg (2.62mmol) DCC prepare 1198mg (91.5%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 579mg (48%).R
f=0.52 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)607[M+H]
+;Mp63-64℃;
(c=0.55,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.862(s,1H),8.236(d,J=7.8Hz,1H),7.442(d,J=7.5Hz,1H),7.282(s,5H),7.249(m,6H),7.009(m,2H),5.085(s,2H),4.620(m,1H),3.850(m,2H),3.702(d,J=14.1Hz,1H),3.426(m,1H),3.132(m,1H),2.926(m,2H),2.444(d,J=13.5Hz,1H),1.919(m,1H),1.819(m,1H),1.335(m,1H),1.200(s,3H),1.150(s,3H),0.852(d,J=6.9Hz,3H),0.821(d,J=6.6Hz,3H).
13C-NMR(DMSO-d
6,75MHz)δ=172.38,171.38,171.33,137.16,136.38,136.03,132.81,129.70,129.62,128.81,128.73,128.60,128.45,128.31,127.12,127.05,126.74,121.04,118.93,117.95,111.37,106.37,79.22,66.68,57.49,56.42,56.32,53.76,41.70,38.84,37.13,25.27,24.84,24.06,22.85,22.79,18.14。
The two benzyl esters (6g) of embodiment 12 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetylglutamate
According to the operation of preparation 6a, by 363mg (0.98mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 593mg (1.19mmol) TosHGlu-OBzl
2, 134mg (0.99mmol) HOBt, 250mg (1.21mmol) DCC prepares 627mg (94.0%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 308mg (49%).R
f=0.52 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)679[M+H]
+;Mp133-134℃;
(c=0.50,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.873(s,1H),8.213(d,J=9Hz,1H),7.408(d,J=7.5Hz,1H),7.324(s,11H),7.005(m,2H),5.097(s,2H),5.073(s,2H),4.414(m,1H),3.950(m,2H),3.742(d,J=13.5Hz,1H),3.638(s,1H),3.433(m,1H),2.908(d,J=12Hz,1H),2.430(t,2H),2.050(m,3H),1.702(m,1H),1.529(m,1H),1.345(s,3H),1.252(s,3H),0.905(m,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.37,172.22,171.55,171.41,136.52,136.40,136.14,132.94,132.85,128.85,128.83,128.51,128.41,128.33,127.12,126.87,121.03,118.91,117.95,111.36,106.39,106.35,79.22,66.70,66.03,57.52,55.81,51.54,41.72,39.06,33.77,29.99,26.38,25.27,24.98,24.92,24.06,23.05,22.72,18.52。
The two benzyl esters (6h) of embodiment 13 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl aspartic acid
According to the operation of preparation 6a, by 374mg (1.01mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 590mg (1.22mmol) TosHAsp-OBzl
2, 141mg (1.04mmol) HOBt, 256mg (1.24mmol) DCC prepares 619mg (92.2%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 285mg (46%).R
f=0.52 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)665[M+H]
+;Mp127–128℃;
(c=0.50,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.884(s,1H),8.302(d,J=8.1Hz,1H),7.412(d,J=7.5Hz,1H),7.335(d,J=5.7Hz,1H),7.325(s,5H),7.295(s,5H),7.005(m,2H),5.068(s,2H),4.784(m,1H),3.929(m,2H),3.748(d,J=13.8Hz,1H),3.445(m,1H),2.904(d,J=6.6Hz,3H),2.581(d,J=12Hz,1H),2.026(m,1H),1.921(m,1H),1.529(m,1H),1.326(s,3H),1.254(s,3H),0.899(s,3H),0.877(s,3H).
13C-NMR(DMSO-d
6,75MHz)δ=172.25,171.23,170.57,170.42,136.56,136.41,136.18,135.95,132.84,128.85,128.79,128.52,128.49,128.39,128.34,128.25,127.12,126.87,121.03,118.92,117.91,111.38,106.42,79.22,66.96,66.41,57.43,55.93,49.09,48.98,41.75,38.86,36.08,25.27,24.99,24.01,23.01,22.79,18.41。
Embodiment 14 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetylserine benzyl esters (6i)
According to the operation of preparation 6a, by 410mg (1.11mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 310mg (1.35mmol) HClSer-OBzl, 154mg (1.14mmol) HOBt, 284mg (1.38mmol) DCC prepares 538mg (89.2%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 215mg (40%).R
f=0.49 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)547[M+H]
+;Mp93-94℃;
(c=0.50,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.874(s,1H),8.134,(d,J=7.8Hz,1H),7.438(d,J=7.5Hz,1H),7.320(s,5H),7.288(m,1H),7.004(m,2H),5.184(t,1H),5.101(s,2H),4.441(m,1H),3.948,(m,2H),3.788(m,2H),3.633(m,1H),3.475(m,1H),2.913(d,J=13.2Hz,1H),2.539(m,1H),2.090(s,1H),1.946(m,1H),1.561(m,1H),1.367(s,3H),1.286(s,3H),0.929(s,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.34,171.56,170.64,136.55,136.27,133.01,128.78,128.37,128.18,127.16,121.02,118.90,117.95,111.42,106.46,79.26,66.50,61.66,57.53,56.42,54.89,41.79,38.85,31.15,25.35,25.02,24.18,23.12,23.00,22.74,18.46。
Embodiment 15 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl threonine benzyl esters (6j)
According to the operation of preparation 6a, by 418mg (1.13mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 338mg (1.38mmol) HClThr-OBzl, 155mg (1.15mmol) HOBt, 286mg (1.39mmol) DCC prepares 570mg (90.0%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 240mg (42%).R
f=0.49 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)561[M+H]
+;Mp93-94℃;
(c=0.65,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.882(s,1H),7.843(d,J=8.4Hz,1H),7.424(d,J=7.5Hz,1H),7.308(s,5H),7.287(m,1H),7.104(m,2H),5.189(s,1H),5.084(t,2H),4.335(d,J=8.4Hz,1H),4.243(s,1H),4.058(m,1H),3.912(m,1H),3.770(d,J=13.5Hz,1H),3.487(d,J=3.9Hz,1H),2.912(d,J=12.9Hz,1H),2.129(m,1H),2.033(m,1H),1.995(s,1H),1.594(m,1H),1.373(s,3H),1.298(s,3H),1.008(d,J=5.7Hz,3H),0.940(s,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.38,171.89,170.78,136.56,136.26,132.99,128.76,128.35,128.24,127.16,121.02,118.90,117.96.111.43,106.51,79.20,66.54,66.50,60.22,57.93,57.53,56.64,41.85,38.83,25.27,25.07,24.17,23.11,22.77,20.67,18.47,14.55。
Embodiment 16 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl-l-lysine benzyl esters (6k)
According to the operation of preparation 6a, by 373mg (1.01mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 614mg (1.65mmol) HClLys (Boc)-OBzl, 143mg (1.06mmol) HOBt, 253mg (1.23mmol) DCC is prepared into 611mg[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-2-(methyl-propyl)-acetyl-N-Boc-Methionin thick product of benzyl ester (88.0%), it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carry out purifying (chloroform: methyl alcohol=200:1), obtain 281mg (46%) pure [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl-N-Boc-Methionin benzyl ester.Rf=0.49 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)688[M+H]+;1H-NMR(DMSO-d6,300MHz)δ=10.862(s,1H),8.181(d,J=7.5Hz,1H),7.429(d,J=7.8Hz,7.309(s,6H),7.002(m,2H),6.741(t,1H),5.092(s,2H),4.308(d,J=4.2Hz,1H),3.937(d,J=12.6Hz,2H),3.756(d,J=13.8Hz,1H),3.462(m,1H),2.938(s,1H),2.868(d,J=6.3Hz,2H),2.072(m,1H),1.919(m,1H),1.720(m,2H),1.529(m,2H),1.362(s,15H),1.272(s,3H),0.916(t,6H).13C-NMR(DMSO-d6,75MHz)δ=172.35,171.97,171.50,171.42,156.01,136.55,136.39,136.20,133.01,132.84,128.82,128.47,128.36,127.16,127.12,121.02,118.90,117.97,111.35,106.36,79.29,77.81,66.53,57.53,56.19,52.34,52.26,41.73,39.03,31.00,29.38,28.73,25.36,25.00,24.13,23.04,22.84,22.77,18.42。
By sterling 250mg (0.36mmol) [tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-2-(methyl-propyl)-acetyl-N-Boc-Methionin benzyl ester is placed in 50mg eggplant bottle, drip 2mL anhydrous ethyl acetate stirring and dissolving, under condition of ice bath, slowly in reaction flask, drip 2.5mL4N HCl/EtOAc, add drying tube.Reacting after 2 hours utilizes TLC chromatography to track to the disappearance of raw material point, stopped reaction.By reaction solution water pump decompressing and extracting, add anhydrous ethyl acetate and fully stir rear water pump decompressing and extracting, three times repeatedly.Add anhydrous diethyl ether and fully stir rear water pump decompressing and extracting, three times repeatedly.Obtain the pure title compound of 202mg (94.6%), it is pale yellow powder.R
f=0.49 (chloroform: methyl alcohol: Glacial acetic acid=20:1:0.10).ESI-MS(m/e)588[M+H]
+;Mp93-94℃;
(c=0.75,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=8.422(d,J=7.2Hz,1H),8.025(s,2H),7.519(d,J=7.5Hz,1H),7.372(d,J=12.3Hz,1H),7.351(s,5H),7.067(m,2H),5.097(s,2H),4.482(m,1H),4.325(d,J=5.1Hz,2H),4.105(m,1H),3.146(m,2H),2.719(s,2H),1.988(m,2H),1.915(s,2H),1.818(m,4H),1.641(m,2H),1.547(s,4H),1.366(s,3H),0.917(t,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.40,171.90,170.41,136.96,136.80,136.29,128.87,128.44,128.34,126.44,121.97,119.36,118.49,111.78,105.88,66.51,57.64,55.11,52.67,52.51,42.42,38.68,38.57,33.73,29.93,26.59,25.19,23.17,22.77,22.64,22.50,21.46,19.75,18.96。
Embodiment 17 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-aceglutamide benzyl esters (6l)
According to the operation of preparation 6a, by 331mg (0.90mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 309mg (1.13mmol) HClGln-OBzl, 122mg (0.90mmol) HOBt, 233mg (1.13mmol) DCC prepares 431mg (82.4%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=100:1), obtain the pure title compound of 108mg (25%).R
f=0.23 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)588[M+H]
+;Mp99-100℃;
(c=0.55,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.872(s,1H),8.246(d,J=7.2Hz,1H),7.428(d,J=7.5Hz,1H),7.320(s,5H),7.307(d,J=7.5Hz,2H),7.002(m,2H),6.789(s,1H),5.093(s,2H),4.336(d,J=4.5Hz,1H),3.961(m,2H),3.752(d,J=13.8Hz,1H),3.450(m,1H),2.912(d,J=12Hz,1H),2.562(d,J=16.5Hz,1H),2.134(m,2H),2.058(m,2H),1.869(m,2H),1.521(m,1H),1.359(s,3H),1.270(s,3H),0.910(t,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.48,172.29,171.83,171.42,136.43,136.23,132.85,128.83,128.46,128.36,127.14,121.02,118.90,117.97,111.36,106.42,79.62,79.28,66.56,57.53,55.96,52.26,52.16,41.72,39.09,31.30,27.08,25.36,25.01,24.11,23.07,22.75,18.47。
Embodiment 18 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl l-asparagine benzyl esters (6m)
According to the operation of preparation 6a, by 495mg (1.34mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 416mg (1.61mmol) HClAsn-OBzl, 186mg (1.38mmol) HOBt, 333mg (1.62mmol) DCC prepares 653mg (85.6%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=100:1), obtain the pure title compound of 170mg (25%).R
f=0.23 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)574[M+H]
+;Mp110-111℃;
(c=0.75,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=8.193(d,J=8.1Hz,1H),7.422(d,J=8.4Hz,2H),7.305(s,5H),7.288(d,J=10.2Hz,1H),7.006(m,3H),5.068(s,2H),3.907(m,2H),3.746(d,J=13.8Hz,1H),3.422(m,1H),3.370(m,2H),2.884(m,1H),2.635(m,2H),2.061(m,1H),1.904(m,1H),1.555(m,1H),1.353(s,3H),1.262(s,3H),0.906(m,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.09,171.48,171.33,171.00,136.58,136.39,136.26,132.98,132.84,128.79,128.37,128.16,127.13,121.01,118.88,117.97,111.38,106.56,79.15,66.56,57.58,55.92,49.12,41.81,38.91,36.69,25.24,25.01,23.99,23.09,22.77,18.43。
Embodiment 19 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-Thiomedon benzyl esters (6n)
According to the operation of preparation 6a, by 450mg (1.22mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 603mg (1.47mmol) TosHMet-OBzl, 167mg (1.24mmol) HOBt, 307mg (1.49mmol) DCC prepares 665mg (93.1%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 300mg (45%).R
f=0.53 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)591[M+H]
+;Mp110-111℃;
(c=0.60,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.886(s,1H),8.225(d,J=7.8Hz,1H),7.424(d,J=7.5Hz,1H),7.327(s,5H),7.297(d,J=18Hz,1H),7.001(m,2H),5.113(m,2H),4.496(m,1H),3.964(m,2H),3.748(d,J=13.5Hz,1H),3.454(m,1H),2.910(m,1H),2.450(m,2H),2.061(m,1H),2.036(m,1H),2.003(s,3H),1,902(m,2H),1.530(m,1H),1.367(s,3H),1.267(s,3H),0.896(m,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.19,171.72,171.40,136.53,136.18,133.02,128.85,128.54,128.47,127.14,121.02,118.91,117.96,111.42,106.39,79.23,66.69,57.50,55.80,51.38,41.76,30.69,29.74,25.30,24.99,24.09,23.07,22.77,18.44,14.94。
Embodiment 20 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetylcysteine benzyl esters (6o)
According to the operation of preparation 6a, by 435mg (1.18mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 389mg (1.476mmol) H-Cys (But)-OBzl, 171mg (1.27mmol) HOBt, 294mg (1.43mmol) DCC prepares 670mg (92.1%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 310mg (46%).R
f=0.52 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)619[M+H]
+;Mp90-91℃;
(c=0.50,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.884(s,1H),8.276(d,J=7.2Hz,1H),7.418(d,J=7.5Hz,1H),7.333(s,5H),7.305(d,J=8.4Hz,1H),(7.006(m,2H),5.117(s,2H),4.595(d,J=5.7Hz,1H),3.942(m,2H),3.758(d,J=13.5Hz,1H),3.465(d,J=7.8Hz,1H),2.916(m,2H),2.009(m,2H),1.724(d,J=9.6Hz,1H),1.560(m,2H),1.366(s,3H),1.251(s,12H),0.920(d,J=4.2Hz,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.35,171.44,170.49,136.55,136.04,133.01,128.80,128.49,128.38,127.16,127.07,126.84,121.01,118.91,117.90,111.43,106.42,79.27,66.83,57.54,56.40,52.74,42.67,40.81,38.89,33.83,30.99,29.92,25.79,25.42,24.94,24.12,22.95,22.79,18.42。
Embodiment 21 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyltryptophan benzyl esters (6p)
According to the operation of preparation 6a, by 786mg (2.13mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 942mg (2.57mmol) HClTrp-OBzl, 291mg (2.15mmol) HOBt, 544mg (2.64mmol) DCC prepares 1161mg (84.5%) title compound, and it is pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 766mg (66%).R
f=0.52 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)646[M+H]
+;Mp101-102℃;
(c=0.85,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.945(s,1H),10.860(s,1H),8.228(d,J=7.5Hz,1H),7.482(m,2H),7.349(m,2H),7.246(s,5H),7.195(m,1H),7.030(m,4H),5.032(s,2H),4.644(m,1H),3.868(m,2H),3.671(t,1H),3.404(m,2H),3.170(m,2H),2.910(d,J=12.3Hz,1H),2.430(d,J=13.2Hz,1H),1.961(m,1H),1.831(m,1H),1.230(s,3H),1.115(s,3H),0.849(m,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.35,171.79,171.34,136.65,136.56,136.07,132.95,128.76,128.39,128.27,127.51,127.20,124.39,121.43,121.01,118.89,118.48,118.02,113.98,111.84,111.40,109.29,106.49,79.23,66.59,57.49,56.30,53.46,41.73,38.91,27.57,25.31,24.93,24.14,22.92,22.72,18.14。
Embodiment 22 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-acetyl proline(Pro) benzyl esters (6q)
According to the operation of preparation 6a, by 700mg (1.90mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 377mg (2.28mmol) HClPro-OBzl, 257mg (1.90mmol) HOBt, 470mg (2.28mmol) DCC prepares 1010mg (90.1%) title compound, pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 500mg (49%).R
f=0.51 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)557[M+H]
+;Mp106-107℃;
(c=0.60,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.861(s,1H),7.376(s,5H),7.364(m,2H),6.999(m,2H),5.128(m,2H),4.907(m,1H),4.361(m,1H),3.953(d,J=13.5Hz,1H),3.713(d,J=13.8Hz,1H),3.555(m,1H),3.472(m,2H),2.958(m,1H),2.478(m,1H),2.209(m,1H),1.963(m,1H),1.892(m,2H),1.710(m,2H),1.625(m,1H),1.463(s,3H),1.189(s,3H),0.862(m,6H).
13C-NMR(DMSO-d
6,75MHz)δ=171.82,171.43,168.03,136.55,136.36,132.99,128.87,128.50,128.27,127.15,121.03,118.91,117.96,111.41,106.34,79.21,66.43,59.58,56.97,50.65,47.01,41.38,33.81,29.14,25.80,25.43,25.11,24.98,24.92,24.31,22.89,22.83,18.19。
Embodiment 23 preparation [tetrahydro-beta-carboline is (2,2-dimethyl-imidazol-4-one-3-yl) also]-2-(methyl-propyl)-Acetyl tyrosine benzyl esters (6r)
According to the operation of preparation 6a, by 607mg (1.64mmol) 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid (5), 876mg (1.98mmol) TosHTyr-OBzl, 231mg (1.71mmol) HOBt, 409mg (1.98mmol) DCC prepares 878mg (85.8%) title compound, pale yellow powder.Chloroform wet method dress post for this powder, dry method is mixed sample, carries out purifying (chloroform: methyl alcohol=200:1), obtain the pure title compound of 410mg (47%).R
f=0.52 (chloroform: methyl alcohol=20:1).ESI-MS(m/e)623[M+H]
+;Mp103-104℃;
(c=0.60,CH
3OH);
1H-NMR(DMSO-d
6,300MHz)δ=10.846(s,1H),9.203(s,1H),8.134(d,J=8.1Hz,1H),7.442(d,J=7.5Hz,1H),7.322(s,1H),7.275(s,5H),6.994(m,4H),6.634(d,J=8.4Hz,2H),5.069(s,2H),4.526(m,1H),3.831(m,2H),3.719(d,J=13.8Hz,1H),3.430(m,1H),2.988(m,1H),2.896(m,1H),2.772(m,1H),2.447(d,J=12.6Hz,1H),1.868(m,1H),1.388(m,2H),1.240(s,3H),1.171(s,3H),0.858(t,6H).
13C-NMR(DMSO-d
6,75MHz)δ=172.40,171.44,171.23,156.45,136.39,136.10,136.05,132.95,132.81,132.63,130.56,128.80,128.43,128.31,127.13,127.00,121.04,118.92,118.00,115.44,111.36,106.40,79.25,66.60,57.44,56.38,54.05,53.97,41.72,38.90,36.47,31.14,25.32,24.89,24.10,22.82,18.14。
Experimental example 16a-r anti-tumour cell proliferative activity is evaluated
1) given the test agent
6a-r of the present invention is all mixed with desired concn with the PBS containing 1% dimethyl sulfoxide (DMSO) (DMSO).
2) cell strain
S180 (mice sarcoma cell), HL60 (human promyelocytic leukemia), K562 (human chronic polymorpho nuclear leukemia cells), H22 (murine hepatocarcinoma cell), SH-sy5y (human neuroblastoma cell) 5 strain tumour cells are all purchased from USS type culture collection institute (ATCC).
3) key instrument and material
Quartzy automatic dual pure water distiller: 1018-B, Jiangsu high honour instrument manufacturing company limited;
High-pressure sterilizing pot: 400Ep-Z ，H+P company;
Water-bath: YLE-1000, Beijing De Tianyou development in science and technology company limited;
Super clean bench: VS-1300-U clean bench, SuZhou Antai Air Tech Co., Ltd.;
Cell incubator: INC153, east, the Five continents, Beijing development in science and technology company limited;
Refrigerated centrifuge: SPD111V, Thermo company;
Microplate reader: MULTISJKAN MK3， Thermo company;
Oscillator plate: MS2Minishaker ，IKA company;
96 porocyte culture plates, 25cm
2culturing bottle: Corning Costar company;
2mL cryopreservation tube: Corning Costar company;
0.22 μ m, 0.45 μ m millipore filtration: upper Haixing County sub-scavenging material factory.
4) main agents
RPMI-1640 culture medium dry powder: Gibco company;
PBS damping fluid: contain NaCl8.2g, KCl0.2g, Na in every liter of solution
2hPO
4h
2o1.56g, KH
2pO
40.2g, pH value 7.4;
Foetal calf serum: Hyclone company;
0.25% pancreatin solution: Hyclone company;
Penicillin, Streptomycin sulphate: solarbio company;
MTT (four tetrazolium bromides): solarbio company, be dissolved in PBS solution, make the solution of 5mg/mL, after filtration sterilization, use, keep in Dark Place;
Zorubicin (ADR): Beijing Hua Feng United Technologies Corp..
DMSO: DMSO, Hyclone company;
5) test method
Growth conditions is good, in the SH-sy5y of logarithmic phase cell by 3 * 10
4the density of individual/mL is inoculated in 96 orifice plate ，Mei hole 100 μ l.Be placed in 37 ℃, 5%CO
2in incubator, cultivate and treat adherent in 4 hours.Growth conditions is good, S180, HL60 in logarithmic phase, K562, H22 cell are according to 4 * 10 respectively
4the density of individual/mL is inoculated in 96 orifice plate ，Mei hole 100 μ l.5 strain cells by default concentration gradient add to be measured, through the 6a-r of sterilising treatment ，Mei hole 25 μ l, control wells adds isopyknic PBS.At 37 ℃, 5%CO
2in incubator, cultivate 48 hours ，Mei holes and add the MTT solution that 25 μ l concentration are 5mg/mL, continue to be placed in 37 ℃, 5%CO
2in incubator, cultivate 4 hours.Centrifugal 4 minutes (3000rpm/min).Careful sucking-off supernatant liquor ，Mei hole adds 120 μ l DMSO dissolve purple residues (first a ceremonial jade-ladle, used in libation), and Oscillating Flat makes to precipitate whole dissolvings for 10 minutes, measures O.D. value (absorbancy), wavelength 570nm in 570nm microplate reader.
According to formula " relative survival rate=(D pastille-D blank)/(D contrast-D blank) * 100% ", calculate sample under each sample concentration inhibiting rate to tumour cell.
Test parallel repetition 3 times, with inhibiting rate, compound concentration is mapped, calculate the IC of the compounds of this invention (6a-r)
50(half effective inhibition concentration) value.Experimental result is as shown in table 1.
Table 16a-r anti-tumour cell proliferative activity (IC
50± SD μ M)
a
A) n=9b) inhibiting rate of this compound to this kind of tumour cell during 100 μ M
Experimental example 26a-r anti-tumor in vivo activity rating
1) experiment material
All compound 6a-r that test-compound: embodiment of the present invention 6-23 obtains;
Positive control is Zorubicin;
Laboratory animal: ICR male mice (clean level), body weight 20 ±2g，You Beijing Vital River Experimental Animals Technology Co., Ltd. provide.One group of every 12 mouse, each group of blank and positive control;
Knurl source: experimentation on animals center, mouse S 180 sarcoma ，You Department Of Medicine, Peking University provides, and goes down to posterity and maintains voluntarily;
Solvent: physiological saline.
2) dosage setting
The dosage of 6a-r and Zorubicin is 1 μ mol/kg, all adopts abdominal cavity single-dose.
3) medicine preparation
6a-r adds a small amount of tween 80 hydrotropy, adds gradually physiological saline to desired concn.Zorubicin is normal saline solution.
4) dosage and administration setting
6a-r Jun Yi abdominal cavity single-dose, by the dosage of 1 μ mol/kg once a day, the every mouse of 0.2mL, successive administration 7 days, administration is 7 times altogether.
Physiological saline contrasts with isopyknic corresponding solution ，Jun Yi abdominal cavity single-dose, the every mouse of 0.2mL, and successive administration 7 days, administration is 7 times altogether.
Zorubicin Jun Yi abdominal cavity single-dose, by the dosage of 1 μ mol/kg once a day, the every mouse of 0.2mL, successive administration 7 days, administration is 7 times altogether.
5) foundation of animal model
Adopt anti-tumor in vivo armpit subcutaneous vaccination model: eugonic S180 ascitic tumor knurl liquid after extraction inoculation 7d under aseptic condition, with normal saline dilution, become the liquid of (1:3) fully to mix, by freshly prepared 0.2% Trypan Blue for tumour cell suspension, after mixing, by white blood cell count(WBC) method, count, dye blue person for dead cell, tinter is not viable cell, and is calculated as follows cell concn and cell survival rate.
Cell concn=(viable count/4 in 4 large grids) * 10
4extension rate=cell count/mL
Cell survival rate=viable count/(viable count+dead cell number) * 100%
Survival rate is greater than to 90% knurl liquid and is prepared into 1.5 * 10 by homogenate method
7the cell suspension of individual/mL, in mouse armpit subcutaneous vaccination 0.2mL/ only, causes solid tumor animal model.
6) mensuration of solid tumor tumour inhibiting rate and the mensuration of spleen index
Experiment proceeds to the 8th day, claims Mouse Weight, and de-cervical vertebra is put to death mouse, and cuts open and get tumour and each main organs of respectively organizing mouse and weigh, and finally statistics is respectively organized the tumour inhibiting rate of medicine.The curative effect of solid tumor represents with the heavy inhibition percentage of knurl, is calculated as follows: the heavy inhibiting rate %=of knurl (heavy/blank group knurl of 1-administration group knurl weight) * 100%.Spleen index=spleen heavy (mg)/execution body weight (g).
7) statistical method
This experimental data statistics all adopts t check and variance analysis.
6a-r is as shown in table 2 to the active result of the anti-tumor in vivo of lotus S180 sarcoma mouse.
The data of table 2 show that 6a-r nearly all shows antitumor action under 1 μ mol/kg dosage, and wherein the antitumor action of 6d and 6h is the strongest.
Table 2.5 and the 6a-r impact on lotus S180 mouse tumor weight
a
Note: a) n=12; *) compare P < 0.001 with physiological saline group
Most antitumor drugs, in killing tumor cell, also have a great impact immunologic function, often cause that spleen index significantly reduces.From spleen index, with blank comparison, under 1 μ mol/kg dosage, Zorubicin causes that spleen index reduces by 50%, 6a-r and do not cause that spleen index reduces.
Claims (3)
1. the compound of a following general formula
Wherein R is CH
3, H, CH
2cH (CH
3)
2, CH (CH
3) CH
2cH
3, CH (CH
3)
2, CH
2c
6h
5, CH
2cH
2cO
2bzl, CH
2cO
2bzl, CH
2oH, CHOHCH
3, (CH
2)
4nH
2, (CH
2)
2cONH
2, CH
2cONH
2, (CH
2)
2sCH
3, CH
2sH, Indole-3-yl-CH
2or CH
2-C
6h
4-OH-p.
2. a method of preparing compound described in claim 1, it is characterized in that, 2-[tetrahydro-beta-carboline also (2,2-dimethyl-imidazol-4-one-3-yl)]-4-methyl-valeric acid, under I-hydroxybenzotriazole, dicyclohexylcarbodiimide and N-methylmorpholine exist, reacts with hydrochloride or the sulfonate of amino-acid benzyl ester under condition of ice bath.
Described in claim 1 compound as the application of preparing antitumor drug.
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| CN106349341A (en) * | 2015-07-13 | 2017-01-25 | 首都医科大学 | Pyridino-indolo-imidazole ketone butyryl-Asp(glucosamine)-OBzl, as well as preparation, activity and application thereof |
| CN106349338A (en) * | 2015-07-13 | 2017-01-25 | 首都医科大学 | Pyridino-indolo-imidazole ketone butyryl-Thr-Phe-glucosamine, as well as preparation, activity and application thereof |
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