CN102875549B - 7-virtue (first) amido-5-amido-6-7-azaindole derivatives and preparation and purposes - Google Patents

7-virtue (first) amido-5-amido-6-7-azaindole derivatives and preparation and purposes Download PDF

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CN102875549B
CN102875549B CN201210340355.6A CN201210340355A CN102875549B CN 102875549 B CN102875549 B CN 102875549B CN 201210340355 A CN201210340355 A CN 201210340355A CN 102875549 B CN102875549 B CN 102875549B
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carbamate
base
pyrrolo
piperidin
pyridine
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CN102875549A (en
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刘滔
胡永洲
董晓武
严晶颖
何俏军
杨波
罗沛华
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Zhejiang University ZJU
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Zhejiang University ZJU
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Abstract

The invention provides class 7-virtue (first) amido-5-amido-6-7-azaindole derivatives and a pharmaceutical salts thereof.React with 2,6-dichloro-3-nitropyridine by corresponding aminomethyl virtue heterocycle, and protect via Boc, be catalyzer with Dimethylamino pyridine in tetrahydrofuran solution, react with tert-Butyl dicarbonate under room temperature after 9 hours and obtain.Synthetic method desired raw material provided by the invention is easy to get, and it is convenient to prepare, and is suitable for suitability for industrialized production.Pharmacological evaluation finds that they are to various tumor cell strains, comprise human colon cancer cell, human leukemia cell, human lung carcinoma cell, Proliferation of Human Ovarian Cell etc. all have good vitro inhibition proliferation function, generally show better vitro inhibition proliferation function and preliminary Inhibiting enzyme activity, can the application in antitumor drug prepared.The general structure of described derivative is as follows.

Description

7-virtue (first) amido-5-amido-6-7-azaindole derivatives and preparation and purposes
Technical field
The invention belongs to organic compound preparation, relate generally to 7-virtue (first) amido-5-amido-6-7-azaindole derivatives and preparation method thereof and the application in antitumor drug.
Background technology
The health of cell cycle is run, and depends on the precision monitoring that perfect cell cycle completes in turn to numerous cell cycle events, thus advances stable running and the conversion in order of cell cycle each phase.Wherein, as the core regulation and control person of cell cycle, cyclin-dependent kinase (cyclin dependent kinases, CDKs) is shown one's talent in the protein kinase of numerous participation cell cycle regulating, is subject to the close attention of antitumor drug research staff; Mammals experiment simultaneously shows, interkinesis CDKs(CDK2, CDK4 and CDK6) between there is the complementary phenomenon of compensatory, and wherein only having CDK1 in Cycle Regulation, show functional uniqueness to a certain degree and irreplaceability, it is mainly through forming mixture with Cyclin B protein binding and then regulating G in the cell cycle 2the carrying out of phase and the transition to M phase (m period).In addition, recent research proves, to the kinase whose suppression of CDK1 by effective inducing apoptosis of tumour cell, only causes the Normocellular cycle arrest of reversibility in short-term simultaneously.Above-mentioned multinomial result of study all effectively confirms that CDK1 kinases will show significant advantage as cancer target in raising drug selectivity, reduction toxic side effect etc.The CDK1 kinases of overexpression in tumour cell is suppressed, thus effectively containment tumor cell proliferation will become the ideas of cancer therapy of a great potential.
Up to now, the CDK1 micromolecular inhibitor reported in domestic and international research is all ATP competitive inhibitor, and as one of CDK1 inhibitor occurred the earliest, purines derivative is the study hotspot of design novel potent CDK micromolecular inhibitor always, larger proportion is occupied in the CDK micromolecular inhibitor structure type entering II/III clinical trial phase and preclinical test at present, being subject to the extensive concern of research staff, is one of CDK1 inhibitor structure type of great exploitation potential for its.
As entered purines CDK micromolecular inhibitor R-roscovitine(Seliciclib or CYC202 of phase II clinical trials at present, CDK1/cyclin B IC 50=0.45 μM, CDK2/cyclin E IC 50=0.7 μM) and just entered at present the purines CDK micromolecular inhibitor SCH 727965(Figure 1-1 that the clinical II phase studies, CDK1/cyclin B IC 50=3 nM, CDK2/cyclin E IC 50=1 nM) etc., all show stronger inhibition tumor cell proliferation activity and significant CDK1 kinase inhibiting activity, for purine analogue provides solid foundation and new direction in the follow-up study in this field.
Summary of the invention
First object of the present invention is to provide 7-virtue (first) amido-5-amido-6-7-azaindole derivatives having no bibliographical information and the pharmaceutical salts thereof of a class formation novelty.Described derivative has following general structure:
Wherein:
Ar be without replacing, mono-substituted phenyl ring or fragrant heterocycle, the substituting group wherein on monosubstituted phenyl ring be fluorine, chlorine, bromine, nitro, containing the alkoxyl group of the straight or branched of 1 to 3 carbon atom and 2-pyridine, 3-pyridine and 4-pyridine ring; Virtue heterocycle selects pyridine, pyrimidine, thiophene, furans or pyrroles;
N is 0 or 1;
R 1with R 2identical or different, select hydrogen, containing the alkyl of the straight or branched of 1 to 3 carbon atom and hydroxyalkyl; Or NR 1r 2for the six-ring replaced, its six-ring can be piperidines, morpholine or piperazine.
Second object of the present invention is to provide the preparation method of 7-virtue (first) amido-5-amido-6-7-azaindole derivatives and pharmaceutical salts thereof, is realized by following steps:
Synthetic route prepared by the present invention:
Target product IV can be obtained according to above-mentioned reaction.Wherein chemical compounds I can according to literature method (Schmid; S. et al; Org Biomol Chem. 2005; 3; 3408.), reacted with 2,6-dichloro-3-nitropyridines by corresponding aminomethyl virtue heterocycle (amino aryl heterocycle); and obtain afterwards via Boc protection (in tetrahydrofuran solution be catalyzer with Dimethylamino pyridine, react 9 hours with tert-Butyl dicarbonate under room temperature).Its reaction is as described below:
(1) in polar solvent, under alkaline condition and 50-80 DEG C of temperature of reaction, there is aminated reacting generating compound II in chemical compounds I and corresponding aminated compounds, products therefrom can obtain sterling through column chromatography.Polar solvent is DMF, methyl alcohol, ethanol, Virahol and methyl-sulphoxide etc., and alkaline matter can select the one in salt of wormwood, sodium carbonate or sodium bicarbonate.
(2) in the anhydrous tetrahydrofuran solution of compound ii; the tetrahydrofuran solution of vinyl magnesium bromide is dripped under nitrogen protection ,-78 DEG C of temperature of reaction; and under-20 DEG C of conditions, react 16-24 hour generation compound III; the ammonium chloride solution adding 20% after reaction terminates in the solution carries out cancellation, and products therefrom can obtain sterling through column chromatography.
(3) compound III removes Boc protecting group, reaction times 12-24 hour in the saturated ethyl acetate solution of hydrochloric acid, generates the dihydrochloride of compounds Ⅳ, can obtain target compound sterling through suction filtration.
3rd object of the present invention is to provide 7-virtue (first) amido-5-amido-6-7-azaindole derivatives and pharmaceutical salts is preparing the application in antitumor drug.Preliminary pharmacological evaluation finds that they are to various tumor cell strains, comprise human colon cancer cell (HCT116), human leukemia cell (HL60), human lung carcinoma cell (A549), Proliferation of Human Ovarian Cell (A2780) etc. all has good vitro inhibition proliferation function, wherein part of compounds has the Inhibit proliferaton effect of highly significant, IC 50reach 10 -2-10 -4μM order of magnitude; In addition, preliminary externally press down enzyme experiment and show, under 10 μMs of concentration, in this compounds, the overwhelming majority has the restraining effect of highly significant to CDK1/Cyclin B, wherein part of compounds (IV a, IV c, IV d, IV e, IV i, IV j) 100%(10 μM is reached to the inhibiting rate of CDK1).
Feature of the present invention is: 7-virtue (first) amido-5-amido-6-7-azaindole derivatives having no bibliographical information and the pharmaceutical salts thereof that construct a class formation novelty, and synthesis desired raw material is easy to get, and it is convenient to prepare, and is suitable for suitability for industrialized production.Meanwhile, preliminary pharmacological activity screening experiment shows, compared with positive control, this compounds generally shows better vitro inhibition proliferation function and preliminary Inhibiting enzyme activity.
Embodiment
The present invention is further described in conjunction with the embodiments.Following embodiment illustrates of the present invention, instead of limits the present invention by any way.
Embodiment 1, t-butylbenzyl (6-piperidines-3-nitropyridine-2-base) carbamate (II a)
By 4g(11.0mmol) t-butylbenzyl (6-chloro-3-nitropyridine-2-base) carbamate is dissolved in the anhydrous DMF of 10ml, adds 1.1g(1.3ml, 13.2mmol in the solution) piperidines and 1.8g (13.2mmol) K 2cO 3, in about 50 DEG C reacting by heating 7h(TLC monitorings).Reaction terminate after, with extraction into ethyl acetate, organic layer through saturated common salt water washing, anhydrous sodium sulfate drying, recycling design.And after through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtain the semi-solid 4.3g of aureus, yield 95%.
1H NMR (500 MHz, DMSO- d 6): δ1.29 (s, 9 H, CH 3×3), 1.48 (m, 4 H, Piperidine-H), 1.61 (m, 2 H, Piperidine-H), 3.58 (m, 4 H, Pip-H), 5.12 (s, 2 H, CH 2), 6.70 (d, 1 H, J= 9.0 Hz, Ar-H), 7.23 (m, 1 H, Ar-H), 7.31 (t, 2 H, J= 7.0 Hz, Ar-H), 7.37 (m, 2 H, Ar-H), 8.13 (d, 1 H, J= 9.0 Hz, Ar-H)。
Embodiment 2, t-butylbenzyl (6-morpholine-3-nitropyridine-2-base) carbamate (II b)
Operating process is identical with embodiment 1, just substitutes piperidines with morpholine, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtains bright yellow solid, yield 95%.
1H NMR (500 MHz, DMSO- d 6): δ1.29 (s, 9 H, CH 3×3), 3.56 (s, 4 H, Morpholine-H), 3.62 (s, 4 H, Morpholine-H), 5.12 (s, 2 H, CH 2), 6.72 (d, 1 H, J= 9.0 Hz, Ar-H), 7.23 (m, 1 H, Ar-H), 7.31 (t, 2 H, J= 7.0 Hz, Ar-H), 7.37 (d, 2 H, J= 7.0 Hz, Ar-H), 8.19 (d, 1 H, J= 9.5 Hz, Ar-H)。
Embodiment 3, t-butylbenzyl (6-diethylin-3-nitropyridine-2-base) carbamate (II c)
Operating process is identical with embodiment 1, just substitutes piperidines with diethylamine, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 9:1), obtains aureus semisolid, yield 90%.
1H NMR (500 MHz, DMSO- d 6): δ0.88 (m, 6 H, CH 3×2), 1.29 (s, 9 H, CH 3×3), 3.38 (m, 4 H, CH 2×2), 5.14 (m, 2 H, CH 2), 6.55 (d, 1 H, J= 9.5 Hz, Ar-H), 7.23 (m, 1 H, Ar-H), 7.32 (t, 2 H, J= 7.0 Hz, Ar-H), 7.37 (m, 2 H, Ar-H), 8.14 (d, 1 H, J= 9.0 Hz, Ar-H)。
Embodiment 4, t-butylbenzyl (6-(2-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate (II d)
Operating process is identical with embodiment 1; just substitute piperidines with 2-piperidine ethanol; obtain aureus semi-solid intermediate; protect via Boc (is catalyzer with Dimethylamino pyridine in tetrahydrofuran solution; 9 hours are reacted with tert-Butyl dicarbonate under room temperature) after; by silica gel column chromatography (eluent: petrol ether/ethyl acetate 9:1), obtain aureus semisolid, yield 93%.
1H NMR (500 MHz, DMSO- d 6): δ1.29 (s, 9 H, OtBu), 1.36 (s, 9 H, CH 3×3), 2.05 (m, 8 H, Piperidine-H, CH 2), 3.41 (m, 3 H, Piperidine-H), 3.89 (m, 2 H, CH 2), 5.11 (s, 2 H, CH 2), 6.66 (d, 1 H, J= 9.5 Hz, Ar-H), 7.22 (t, 1 H, J= 7.5 Hz, Ar-H), 7.30 (t, 2 H, J= 7.5 Hz, Ar-H), 7.36 (d, 2 H, J= 7.0 Hz, Ar-H), 8.12 (d, 1 H, J= 9.0 Hz, Ar-H)。
Embodiment 5, t-butylbenzyl (6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate (II e)
Operating process is identical with embodiment 4, just substitutes 2-piperidine ethanol with 4-piperidine ethanol, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 8:1), obtains aureus semisolid, yield 91%.
1H NMR (500 MHz, DMSO- d 6): δ1.17 (m, 2 H, Piperidine-H), 1.38 (s, 9 H, CH 3×3), 1.52 (s, 9 H, CH 3×3), 1.64 (q, 3 H, J= 6.5 Hz, Piperidine-H), 1.79 (d, 2 H, J= 13.5 Hz, CH 2), 2.90 (m, 2 H, Piperidine-H), 4.15 (m, 2 H, Piperidine-H), 4.30 (d, 2 H, J= 13.0 Hz, CH 2), 5.10 (s, 2 H, CH 2), 6.35 (d, 1 H, J= 9.0 Hz, Ar-H), 7.22 (t, 1 H, J= 7.5 Hz, Ar-H), 7.30 (t, 2 H, J= 7.5 Hz, Ar-H), 7.43 (d, 2 H, J= 7.5 Hz, Ar-H), 8.14 (d, 1 H, J= 9.0 Hz, Ar-H)。
Embodiment 6, t-butylbenzyl (6-(2-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate (II f)
Operating process is identical with embodiment 4, just substitutes 2-piperidine ethanol with 2-piperidine carbinols, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 8:1), obtains aureus semisolid, yield 93%.
1H NMR (500 MHz, DMSO- d 6): δ1.28 (s, 9 H, CH 3×3), 1.38 (s, 9 H, CH 3×3), 1.55 (m, 4 H, Piperidine-H), 1.67 (m, 2 H, Piperidine-H), 3.04 (m, 1 H, Piperidine-H), 4.04 (m, 3 H, Piperidine-H, CH), 4.43 (m, 1 H, CH), 5.10 (s, 2 H, CH 2), 6.68 (d, 1 H, J= 9.0 Hz, Ar-H), 7.22 (t, 1 H, J= 7.5 Hz, Ar-H), 7.35 (m, 4 H, Ar-H), 8.13 (d, 1 H, J= 9.0 Hz, Ar-H)。
Embodiment 7, t-butylbenzyl (6-(4-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate (II g)
Operating process is identical with embodiment 4, just substitutes 2-piperidine ethanol with 4-piperidine carbinols, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtains aureus semisolid, yield 91%.
1H NMR (500 MHz, DMSO- d 6): δ1.27 (m, 2 H, Piperidine-H), 1.38 (s, 9 H, CH 3×3), 1.49 (s, 9 H, CH 3×3), 1.82 (d, 2 H, J= 12.5 Hz, Piperidine-H), 1.98 (m, 1 H, Piperidine-H), 2.91 (t, 2 H, J= 12.0 Hz, Piperidine-H), 3.95 (d, 2 H, J= 6.5 Hz, Piperidine-H), 4.32 (d, 2 H, J= 12.5 Hz, CH 2), 5.10 (s, 2 H, CH 2), 6.53 (d, 1 H, J= 9.0 Hz, Ar-H), 7.22 (t, 1 H, J= 7.5 Hz, Ar-H), 7.30 (t, 2 H, J= 8.0 Hz, Ar-H), 7.42 (d, 2 H, J= 7.5 Hz, Ar-H), 8.15 (d, 1 H, J= 9.5 Hz, Ar-H)。
Embodiment 8, butyl benzyl (6-((2-tertbutyloxycarbonyl oxygen base) ethyl-(N-tertbutyloxycarbonyl) amido)-3-nitropyridine-2-base) carbamate (II h)
Operating process is identical with embodiment 4, just substitutes 2-piperidine ethanol with thanomin, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 9:1), obtains aureus liquid, yield 90%.
1H NMR (500 MHz, DMSO- d 6): δ1.29 (s, 9 H, CH 3×3), 1.32 (s, 9 H, CH 3×3), 1.49 (s, 9 H, CH 3×3), 4.04 (m, 4 H, CH 2), 5.09 (s, 2 H, CH 2), 7.25 (m, 1 H, Ar-H), 7.35 (m, 4 H, Ar-H), 7.75 (d, 1 H, J= 9.0 Hz, Ar-H), 8.41 (d, 1 H, J= 9.0 Hz, Ar-H)。
Embodiment 9, butyl benzyl (6-((2-tertbutyloxycarbonyl oxygen base) ethyl-(N-tertbutyloxycarbonyl) amido)-3-nitropyridine-2-base) carbamate (II i)
Operating process is identical with embodiment 4, just substitutes 2-piperidine ethanol with 2-amino butanol, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 9:1), obtains aureus liquid, yield 89%.
1H NMR (500 MHz, DMSO- d 6): δ0.62 (m, 3 H, CH 3), 1.38 (m, 2 H, CH 2), 1.29 (s, 9 H, CH 3×3), 1.32 (s, 9 H, CH 3×3), 1.45 (s, 9 H, CH 3×3), 4.07 (m, 1 H, CH), 4.31 (m, 1 H, CH), 4.56 (m, 1 H, CH), 5.09 (s, 2 H, CH 2), 7.23 (m, 1 H, Ar-H), 7.32 (m, 4 H, Ar-H), 7.44 (d, 1 H, J= 9.0 Hz, Ar-H), 8.40 (d, 1 H, J= 9.0 Hz, Ar-H)。
Embodiment 10, butyl benzyl (6-bis-(2-tertbutyloxycarbonyl oxygen ethylamine)-3-nitropyridine-2-base) carbamate (II j)
Operating process is identical with embodiment 4, just substitutes 2-piperidine ethanol with diethanolamine, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 9:1), obtains aureus semisolid, yield 93%.
1H NMR (500 MHz, DMSO- d 6): δ1.37 (s, 9 H, CH 3×3), 1.45 (s, 9 H, CH 3×3), 1.47 (s, 9 H, CH 3×3), 3.69 (m, 4 H, CH 2×2), 4.14 (m, 4 H, CH 2×2), 5.19 (s, 2 H, CH 2), 6.43 (d, 1 H, J= 9.0 Hz, Ar-H), 7.21 (t, 1 H, J= 7.5 Hz, Ar-H), 7.30 (t, 2 H, J= 7.5 Hz, Ar-H), 7.37 (d, 2 H, J= 7.5 Hz, Ar-H), 8.17 (d, 1 H, J= 9.5 Hz, Ar-H)。
Embodiment 11, t-butylbenzyl 6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base (pyridine-2-methyl) carbamate (II k)
Operating process is identical with embodiment 4, just substitute t-butylbenzyl (the chloro-3-nitropyridine of 6--2-base) carbamate with tertiary butyl 6-chloro-3-nitropyridine-2-base (pyridine-2-methyl) carbamate, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 2:1), obtain brown color semi-solid, two step yields 75%.
1H NMR (500 MHz, DMSO- d 6): δ1.12 (m, 2 H, Piperidine-H), 1.38 (s, 9 H, CH 3×3), 1.49 (d, 9 H, J= 10.0 Hz, CH 3×3), 1.61 (q, 2 H, J= 6.5 Hz, Piperidine-H), 1.72 (m, 3 H, Piperidine-H, CH 2), 2.82 (t, 2 H, J= 12.5 Hz, Piperidine-H), 4.11 (m, 2 H, CH 2), 4.18 (d, 2 H, J= 13.0 Hz, Piperidine-H), 5.21 (s, 2 H, CH 2), 6.34 (d, 1 H, J= 9.0 Hz, Ar-H), 7.13 (t, 1 H, J= 5.5 Hz, Ar-H), 7.56 (d, 1 H, J= 8.0 H, Ar-H z), 7.63 (t, 1 H, J= 7.5 Hz, Ar-H), 8.15 (d, 1 H, J= 9.5 Hz, Ar-H), 8.50 (d, 1 H, J= 4.5 Hz, Ar-H)。
Embodiment 12, t-butylbenzyl 6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base (pyridine-3-methyl) carbamate (II l)
Operating process is identical with embodiment 4, just substitute t-butylbenzyl (the chloro-3-nitropyridine of 6--2-base) carbamate with tertiary butyl 6-chloro-3-nitropyridine-2-base (pyridine-3-methyl) carbamate, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 1:2), obtain brown color semi-solid, two step yields 60%.
1H NMR (500 MHz, DMSO- d 6): δ1.18 (m, 2 H, Piperidine-H), 1.37 (s, 9 H, CH 3×3), 1.49 (s, 9 H, CH 3×3), 1.64 (q, 2 H, J= 8.0 Hz, Piperidine-H), 1.76 (m, 1 H, Piperidine-H), 1.81 (d, 2 H, J= 16.0 Hz, Piperidine-H), 2.93 (t, 2 H, J= 15.5 Hz, CH 2), 4.14 (t, 2 H, J= 8.0 Hz, CH 2), 4.30 (d, 2 H, J= 10.0 Hz, Piperidine-H), 5.10 (s, 2 H, CH 2), 6.36 (d, 1 H, J= 11.5 Hz, Ar-H), 7.24 (m, 1 H, Ar-H), 7.89 (d, 1 H, J= 8.5 Hz, Ar-H), 8.12 (d, 1 H, J= 11.5 Hz, Ar-H), 8.47 (d, 1 H, J= 5.0 Hz, Ar-H), 8.57 (s, 1 H, Ar-H)。
Embodiment 13, t-butylbenzyl 6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base (pyridine-4-methyl) carbamate (II m)
Operating process is identical with embodiment 4, just substitute t-butylbenzyl (the chloro-3-nitropyridine of 6--2-base) carbamate with tertiary butyl 6-chloro-3-nitropyridine-2-base (pyridine-4-methyl) carbamate, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 1:3), obtain brown color semi-solid, two step yields 40%.
1H NMR (500 MHz, DMSO- d 6): δ1.14 (m, 2 H, Piperidine-H), 1.37 (s, 9 H, CH 3×3), 1.49 (s, 9 H, CH 3×3), 1.62 (q, 2 H, J= 8.5 Hz, Piperidine-H), 1.70 (m, 1 H, Piperidine-H), 1.77 (d, 2 H, J= 16.5 Hz, Piperidine-H), 2.88 (t, 2 H, J= 16.0 Hz, CH 2), 4.13 (t, 2 H, J= 8.5 Hz, CH 2), 4.21 (d, 2 H, J= 16.5 Hz, Piperidine-H), 5.15 (s, 2 H, CH 2), 6.36 (d, 1 H, J= 11.5 Hz, Ar-H), 7.36 (d, 2 H, J= 7.5 Hz, Ar-H), 8.15 (d, 1 H, J= 11.5 Hz, Ar-H), 8.52 (d, 2 H, J= 7.0 Hz, Ar-H)。
Embodiment 14, butyl benzyl (5-(piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III a)
By compound 2g(4.8mmol) II abe dissolved in anhydrous tetrahydro furan 25ml, N 2protection, is cooled to-78 DEG C by solution.Drip 14.6ml(1 Mol/L, 14.6mmol at this temperature) tetrahydrofuran solution of vinyl magnesium bromide, dropwise rear continuation and stir 1 hour at-78 DEG C.Then slowly be warming up to-20 DEG C, react 16 hours at this temperature.Add the ammonium chloride solution 25ml cancellation of 20% after reaction terminates in the solution, reclaim a large amount of solvent.Then with extraction into ethyl acetate, organic layer through saturated common salt water washing, anhydrous sodium sulfate drying, recycling design.And through silica gel column chromatography (eluent: petrol ether/ethyl acetate 6:1), obtain pale yellow semi-solid 190mg, yield 10%.
H NMR (500 MHz, DMSO- d 6): δ1.33 (s, 9 H, CH 3×3), 1.51 (m, 6 H, Piperidine-H), 4.23 (m, 4 H, Piperidine-H), 4.79 (s, 2 H, CH 2), 6.58 (d, 1 H, J= 8.5 Hz, Ar-H), 7.07 (s, 1 H, Ar-H), 7.20 (m, 1 H, Ar-H), 7.27 (t, 3 H, J= 7.5 Hz, Ar-H), 7.39 (m, 3 H, Ar-H)。
Embodiment 15, butyl benzyl (5-morpholine-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III b)
Operating process is identical with embodiment 14, just with compound ii balternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 6:1), obtain pale yellow semi-solid, yield 13%.
1H NMR (500 MHz, DMSO- d 6): δ1.39 (s, 9 H, CH 3×3), 3.30 (t, 4 H, J= 4.5 Hz, Morpholine-H), 3.80 (t, 4 H, J= 4.5 Hz, Morpholine-H), 4.94 (s, 2H, CH 2), 7.04 (d, 1 H, J= 8.5 Hz, Ar-H), 7.23 (t, 1 H, J= 7.5 Hz, Ar-H), 7.30 (m, 3 H, Ar-H), 7.38 (m, 3 H, Ar-H)。
Embodiment 16, butyl benzyl (5-diethylamide-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III c)
Operating process is identical with embodiment 14, just with compound ii calternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtain pale yellow semi-solid, yield 10%.
1H NMR (500 MHz, DMSO- d 6): δ0.81 (m, 6 H, CH 3×2), 1.30 (s, 9 H, CH 3×3), 3.22 (m, 4 H, CH 2×2), 4.79 (s, 2 H, CH 2), 6.34 (d, 1 H, J= 8.5 Hz, Ar-H), 7.06 (d, 1 H, J= 7.5 Hz, Ar-H), 7.20 (m, 1 H, Ar-H), 7.28 (t, 3 H, J= 7.5 Hz, Ar-H), 7.41 (d, 2 H, J= 7.5 Hz, Ar-H)。
Embodiment 17, butyl benzyl (5-(2-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III d)
Operating process is identical with embodiment 14, just with compound ii dalternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 6:1), obtain pale yellow semi-solid, yield 15%.
1H NMR (500 MHz, DMSO- d 6): δ1.33 (s, 9 H, CH 3×3), 1.38 (s, 9 H, CH 3×3), 1.55 (m, 6 H, Piperidine-H, CH 2), 1.80 (m, 2 H, Piperidine-H), 2.80 (m, 1 H, Piperidine-H), 3.95 (m, 2 H, CH 2, Piperidine-H), 4.09 (m, 2 H, CH 2), 4.81 (s, 2 H, CH 2), 6.51 (d, 1 H, J= 8.5 Hz, Ar-H), 7.05 (s, 1 H, Ar-H), 7.19 (m, 1 H, Ar-H), 7.27 (t, 3 H, J= 7.0 Hz, Ar-H), 7.37 (m, 3 H, Ar-H)。
Embodiment 18, butyl benzyl (5-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III e)
Operating process is identical with embodiment 14, just with compound ii ealternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtain pale yellow semi-solid, yield 13%.
1H NMR (500 MHz, DMSO- d 6): δ1.14 (m, 2 H, Piperidine-H),1.33 (s, 9 H, CH 3×3), 1.41 (s, 9 H, CH 3×3), 1.52 (m, 3 H, Piperidine-H), 1.67 (d, 2 H, J= 12.0 Hz, Piperidine-H), 3.94 (d, 2 H, J= 12.0 Hz, Piperidine-H), 4.07 (t, 2 H, J= 6.5 Hz, CH 2), 4.14 (m, 2 H, CH 2), 4.79 (s, 2 H, CH 2), 6.59 (d, 1 H, J= 8.5 Hz, Ar-H), 7.07 (d, 1 H, J= 8.0 Hz, Ar-H), 7.19 (m, 1 H, Ar-H), 7.27 (t, 3 H, J= 7.5 Hz, Ar-H), 7.39 (d, 3 H, J= 8.0 Hz, Ar-H)。
Embodiment 19, butyl benzyl (5-(2-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III f)
Operating process is identical with embodiment 14, just with compound ii falternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 6:1), obtain pale yellow semi-solid, yield 9%.
1H NMR (500 MHz, DMSO- d 6): δ1.32 (s, 9 H, CH 3×3), 1.36 (s, 9 H, CH 3×3), 1.54 (m, 4 H, Piperidine-H), 1.68 (m, 2 H, Piperidine-H), 3.72 (m, 1 H, Piperidine-H), 4.13 (m, 3 H, Piperidine-H, CH), 4.46 (m, 1 H, CH), 4.78 (s, 2 H, CH 2), 6.55 (d, 1 H, J= 8.5 Hz, Ar-H), 7.07 (m, 1 H, Ar-H), 7.19 (m, 1 H, Ar-H), 7.26 (t, 3 H, J= 7.5 Hz, Ar-H), 7.38 (m, 3 H, Ar-H)。
Embodiment 20, butyl benzyl (5-(4-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III g)
Operating process is identical with embodiment 14, just with compound ii galternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 6:1), obtain pale yellow semi-solid, yield 11%.
1H NMR (500 MHz, DMSO- d 6): δ1.28 (m, 2 H, Piperidine-H), 1.40 (s, 9 H, CH 3×3), 1.47 (s, 9 H, CH 3×3), 1.73 (d, 2 H, J= 12.0 Hz, Piperidine-H), 1.83 (m, 1 H, Piperidine-H), 2.71 (t, 2 H, J= 12.0 Hz, Piperidine-H), 3.93 (d, 2 H, J= 6.5 Hz, Piperidine-H), 4.04 (d, 2 H, J= 12.0 Hz, CH 2), 4.61 (s, 2 H, CH 2), 6.66 (d, 1 H, J= 8.5 Hz, Ar-H), 7.23 (m, 1 H, Ar-H), 7.29 (t, 3 H, J= 7.0 Hz, Ar-H), 7.38 (m, 3 H, Ar-H)。
Embodiment 21, butyl benzyl (5-((N-tertbutyloxycarbonyl)-2-tertbutyloxycarbonyl oxygen ethylamine)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III h)
Operating process is identical with embodiment 14, just with compound ii halternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtain pale yellow semi-solid, yield 13%.
1H NMR (500 MHz, DMSO- d 6): δ1.31 (s, 9 H, CH 3×3), 1.34 (s, 9 H, CH 3×3), 1.39 (s, 9 H, CH 3×3), 3.88 (t, 2 H, J= 6.0 Hz, CH 2), 4.06 (t, 2 H, J= 6.0 Hz, CH 2), 5.00 (s, 2 H, CH 2), 6.48 (m, 1 H, Ar-H), 7.18 (m, 1 H, Ar-H), 7.25 (t, 2 H, J= 7.5 Hz, Ar-H), 7.34 (m, 3 H, Ar-H), 7.36 (s, 1 H, Ar-H), 7.56 (m, 1 H, Ar-H)。
Embodiment 22, butyl benzyl (5-(1-tertbutyloxycarbonyl oxygen butane-2-(N-tertbutyloxycarbonyl) is amino)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III i)
Operating process is identical with embodiment 14, just with compound ii ialternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtain pale yellow semi-solid, yield 10%.
1H NMR (500 MHz, DMSO- d 6): δ0.77 (m, 2 H, CH 2), 0.89 (t, 3 H, J= 7.5 Hz, CH 3), 1.35 (s, 9 H, CH 3×3), 1.38 (s, 9 H, CH 3×3), 1.40 (s, 9 H, CH 3×3), 3.99 (m, 1 H, CH), 4.28 (m, 1 H, CH), 4.47 (d, 1 H, J= 5.5 Hz, CH), 4.99 (s, 2 H, CH 2), 6.58 (m, 1 H, Ar-H), 7.18 (m, 1 H, Ar-H), 7.23 (m, 2 H, Ar-H), 7.33 (m, 4 H, Ar-H), 7.57 (m, 1 H, Ar-H)。
Embodiment 23, butyl benzyl (5-bis-(2-tertbutyloxycarbonyl oxygen ethylamine)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate (III j)
Operating process is identical with embodiment 14, just with compound ii jalternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 7:1), obtain pale yellow semi-solid, yield 10%.
1H NMR (500 MHz, DMSO- d 6): δ1.32 (s, 9 H, CH 3×3), 1.38 (s, 18 H, CH 3×6), 3.55 (m, 4 H, CH 2×2), 4.06 (t, 4 H, J= 5.5 Hz, CH 2×2), 4.81 (s, 2 H, CH 2), 6.49 (d, 1 H, J= 8.5 Hz, Ar-H), 7.09 (m, 1 H, Ar-H), 7.19 (m, 1 H, Ar-H), 7.30 (t, 3 H, J= 7.5 Hz, Ar-H), 7.36 (d, 2 H, J= 7.5 Hz, Ar-H)。
Embodiment 24, N-benzyl-5-(piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridine-7-amine hydrochlorate (IV a)
By 60mg(0.15 mmol) compound III abe dissolved in ethyl acetate, drip the ethyl acetate solution that hydrochloric acid is saturated, rise to room temperature under condition of ice bath, stir 12h, suction filtration, a small amount of ethyl acetate washing, obtains red brown solid product 30mg, yield 65%.
1H NMR (500 MHz, DMSO- d 6): δ1.52 (m, 2 H, Piperidine-H), 1.61 (m, 4 H, Piperidine-H), 2.90 (t, 4 H, J= 5 Hz, Piperidine-H), 4.33 (d, 2 H, J= 6.5 Hz, CH 2), 6.00 (m, 2 H, Ar-H), 6.80 (d, 1 H, J= 8.0 Hz, Ar-H), 7.19 (t, 1 H, J= 7.5 Hz, Ar-H), 7.29 (m, 2 H, Ar-H), 7.33 (d, 2 H, J= 7.0 Hz, Ar-H)。
Embodiment 25, N-benzyl-5-morpholine-1H-pyrrolo-[2,3-c] pyridine-7-amine hydrochlorate (IV b)
Operating process is identical with embodiment 24, just with compound III balternative compounds III a, obtain reddish-brown semisolid, yield 61%.
1H NMR (500 MHz, DMSO- d 6): δ3.40 (t, 4 H, J= 4.5 Hz, Morpholine-H), 3.73 (t, 4 H, J= 4.5 Hz, Morpholine-H), 4.66 (s, 2 H, CH 2), 7.25 (t, 1 H, J= 7.5 Hz, Ar-H), 7.33 (t, 2 H, J= 7.5 Hz, Ar-H), 7.39 (d, 1 H, J= 4.0 Hz, Ar-H), 7.04 (m, 4 H, Ar-H)。
Embodiment 26, N 7-benzyl-N 5, N 5-diethyl-1H-pyrrolo-[2,3-c] pyridine-5,7-diamine hydrochloride (IV c)
Operating process is identical with embodiment 24, just with compound III calternative compounds III a, obtain reddish-brown semisolid, yield 69%.
1H NMR (500 MHz, DMSO- d 6): δ0.80 (m, 6 H, CH 3×2), 3.32 (m, 4 H, CH 2×2), 4.60 (s, 2 H, CH 2), 6.87 (s, 1 H, Ar-H), 7.21 (m, 1 H, Ar-H), 7.30 (t, 3 H, J= 7.5 Hz, Ar-H), 7.34 (m, 3 H, Ar-H)。
Embodiment 27,2-(1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-2-yl) ethylate hydrochlorate (IV d)
Operating process is identical with embodiment 24, just with compound III dalternative compounds III a, obtain reddish-brown semisolid, yield 63%.
1H NMR (500 MHz, DMSO- d 6): δ1.98 (m, 8 H, Piperidine-H, CH 2), 3.37 (m, 2 H, Piperidine-H), 3.80 (m, 3 H, Piperidine-H, CH 2), 4.65 (d, 2 H, J= 7.0 Hz, CH 2), 7.05 (m, 1 H, Ar-H), 7.25 (m, 1 H, Ar-H), 7.36 (m, 4 H, Ar-H), 7.41 (m, 4 H, Ar-H)。
Embodiment 28,2-(1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate (IV e)
Operating process is identical with embodiment 24, just with compound III ealternative compounds III a, obtain reddish-brown semisolid, yield 61%.
1H NMR (500 MHz, DMSO- d 6): δ1.61 (m, 3 H, Piperidine-H),1.67 (q, 2 H, J= 6.5 Hz, CH 2), 1.98 (m, 2 H, Piperidine-H), 3.34 (m, 1 H, Piperidine-H), 3.57 (m, 2 H, Piperidine-H), 3.65 (m, 1 H, Piperidine-H), 4.16 (t, 2 H, J= 6.5 Hz, CH 2), 4.66 (s, 2 H, CH 2), 7.24 (m, 1 H, Ar-H), 7.33 (t, 3 H, J= 7.5 Hz, Ar-H), 7.40 (d, 3 H, J= 9.5 Hz, Ar-H), 7.49 (s, 1 H, Ar-H)。
Embodiment 29, (1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-2-yl) methylate hydrochlorate (IV f)
Operating process is identical with embodiment 24, just with compound III falternative compounds III a, obtain reddish-brown semisolid, yield 57%.
1H NMR (500 MHz, DMSO- d 6): δ2.02 (m, 6 H, Piperidine-H), 3.01 (d, 1 H, J= 12.0 Hz, Piperidine-H), 3.21 (d, 1 H, J= 12.0 Hz, Piperidine-H), 3.39 (m, 3 H, Piperidine-H, CH 2), 4.66 (d, 2 H, J= 8.0 Hz, CH 2), 7.24 (t, 1 H, J= 7.5 Hz, Ar-H), 7.33 (t, 3 H, J= 7.5 Hz, Ar-H), 7.40 (d, 3 H, J= 7.5 Hz, Ar-H), 7.46 (s, 1 H, Ar-H)。
Embodiment 30, (1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) methylate hydrochlorate (IV g)
Operating process is identical with embodiment 24, just with compound III galternative compounds III a, obtain reddish-brown semisolid, yield 65%.
1H NMR (500 MHz, DMSO- d 6): δ2.08 (m, 5 H, Piperidine-H), 3.21 (m, 2 H, Piperidine-H), 3.77 (m, 2 H, Piperidine-H), 4.01 (d, 2 H, J= 6.0 Hz, CH 2), 4.67 (s, 2 H, CH 2), 7.21 (m, 1 H, Ar-H), 7.35 (m, 3 H, Ar-H), 7.43 (m, 4 H, Ar-H)。
Embodiment 31,2-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base amine) ethylate hydrochlorate (IV h)
Operating process is identical with embodiment 24, just with compound III halternative compounds III a, obtain reddish-brown semisolid, yield 60%.
1H NMR (500 MHz, DMSO- d 6): δ3.25 (t, 1 H, J= 5.0 Hz, CH), 3.32 (m, 1 H, CH), 3.70 (m, 1 H, CH), 3.76 (t, 1 H, J= 5.0 Hz, CH), 4.69 (s, 2 H, CH 2), 7.23 (m, 1 H, Ar-H), 7.34 (m, 2 H, Ar-H), 7.40 (t, 3 H, J= 7.5 Hz, Ar-H), 7.44 (d, 2 H, J= 7.5 Hz, Ar-H), 7.48 (s, 1 H, Ar-H)。
Embodiment 32,2-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base amine) Kauri-butanol hydrochloric salt (IV i)
Operating process is identical with embodiment 24, just with compound III ialternative compounds III a, obtain reddish-brown semisolid, yield 64%.
1H NMR (500 MHz, DMSO- d 6): δ1.04 (m, 3 H, CH 3), 2.05 (m, 2 H, CH 2), 3.61 (m, 1 H, CH), 3.86 (m, 1 H, CH), 4.19 (m, 1 H, CH), 4.77 (s, 2 H, CH 2), 7.38 (m, 2 H, Ar-H), 7.44 (m, 3 H, Ar-H), 7.50 (m, 3 H, Ar-H)。
Embodiment 33,2,2 '-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base amine) diethylate hydrochlorate (IV j)
Operating process is identical with embodiment 24, just with compound III jalternative compounds III a, obtain reddish-brown semisolid, yield 66%.
1H NMR (500 MHz, DMSO- d 6): δ3.49 (t, 4 H, J= 5.0 Hz, CH 2×2), 3.75 (t, 4 H, J= 5.0 Hz, CH 2×2), 4.65 (s, 2 H), 7.30 (m, 2 H, Ar-H), 7.36 (t, 3 H, J= 7.5 Hz, Ar-H), 7.42 (d, 2 H, J= 7.0 Hz, Ar-H), 7.62 (s, 1 H, Ar-H)。
Embodiment 34,2-(1-(7-(pyridine-2-ylmethyl ammonia)-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate (IV k)
Operating process is identical with embodiment 14, just with compound ii kalternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 2:1), obtain pale yellow semi-solid intermediate; Direct input next step, specific operation process is identical with embodiment 24, just with this intermediate alternative compounds III a, two step yields 11%.
1H NMR (500 MHz, DMSO- d 6): δ1.25 (m, 2 H, Piperidine-H), 2.03 (m, 5 H, Piperidine-H, CH 2), 3.68 (m, 2 H, Piperidine-H), 4.19 (m, 4 H, Piperidine-H, CH 2), 5.18 (s, 2 H, CH 2), 7.40 (d, 1 H, J= 10.5 Hz, Ar-H), 7.82 (d, 1 H, J= 10.0 Hz, Ar-H), 8.04 (m, 1 H, Ar-H), 8.14 (m, 1 H, Ar-H), 8.22 (d, 1 H, J= 7.5 Hz, Ar-H), 8.64 (m, 2 H, Ar-H), 8.84 (d, 1 H, J= 6.0 Hz, Ar-H)。
Embodiment 35,2-(1-(7-(pyridin-3-yl methyl ammonia)-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate (IV l)
Operating process is identical with embodiment 14, just with compound ii lalternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 1:1), obtain pale yellow semi-solid intermediate; Direct input next step, specific operation process is identical with embodiment 24, just with this intermediate alternative compounds III a, two step yields 10%.
1H NMR (500 MHz, DMSO- d 6): δ1.70 (m, 4 H, Piperidine-H), 2.05 (d, 3 H, J= 13.5 Hz, Piperidine-H, CH 2), 3.60 (m, 4 H, Piperidine-H ), 4.10 (m, 2 H, CH 2), 4.45 (s, 2 H, CH 2), 5.26 (m, 2 H, Ar-H), 5.84 (s, 1 H, Ar-H), 8.21 (d, 1 H, J= 9.0 Hz, Ar-H), 8.77 (d, 1 H, J= 10.5 Hz, Ar-H), 8.97 (d, 1 H, J= 7.0 Hz, Ar-H), 9.06 (s, 1 H, Ar-H)。
Embodiment 36,2-(1-(7-(pyridin-4-yl methyl ammonia)-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate (IV m)
Operating process is identical with embodiment 14, just with compound ii malternative compounds II a, through silica gel column chromatography (eluent: petrol ether/ethyl acetate 1:2), obtain the semi-solid intermediate of light brown; Direct input next step, specific operation process is identical with embodiment 24, just with this intermediate alternative compounds III a, two step yields 8%.
1H NMR (500 MHz, DMSO- d 6): δ1.32 (m, 2 H, Piperidine-H), 2.04 (m, 5 H, Piperidine-H, CH 2), 4.16 (m, 6 H, , Piperidine-H, CH 2), 4.55 (s, 2 H, CH 2), 8.11 (m, 2 H Ar-H), 8.18 (d, 2 H, J= 6.0 Hz Ar-H), 8.78 (d, 1 H, J= 6.5 Hz Ar-H), 8.99 (d, 2 H, J= 6.5 Hz Ar-H)。
Embodiment 37, anti-tumor biological testing method:
Tumour cell isolated culture: choose tumour cell HL60, A549, HCT116, A2780 are in 37 DEG C, 5% CO 2hatch in cell culture incubator, go down to posterity when cell density grows to 70-90% (attached cell Puck ' s EDTA goes down to posterity after digesting), for later needed for experiment.
Mtt assay measures 7-virtue (first) amido-5-amido-6-7-azaindole derivatives dihydrochloride to the In-vitro Inhibitory Effect of different knurl strain:
Compound DMSO is dissolved, dilution; To the above-mentioned tumour cell of logarithmic phase be in, with 2 × 10 4individual/mL is inoculated in 96 well culture plates, and every hole adds compound 2 μ l, and final concentration is 12.0 μMs, 6.0 μMs, 3.0 μMs, 1.5 μMs, jointly in 37 DEG C, 5% CO 248 hours are hatched, with DMSO(1% in cell culture incubator) be blank.Every hole adds the MTT solution 20 μ L that concentration is 2.5mg/mL, continues to cultivate 4h, and suck supernatant liquor, every hole adds the DMSO of 100 μ L, shakes up, and under 570nM wavelength, measure the absorbancy (OD value) in each hole by microplate reader, the data obtained is for calculating IC 50.Acquired results is see table 1, and table 1 is that part 7-virtue (first) amido-5-amido-6-7-azaindole derivatives dihydrochloride is active to the vitro inhibition of different tumor cell line.
" nd " represents undetermined IC 50value.
External activity test result as can be seen from table 1:
(1) to comprising HL60(human leukemia cell), A549(human lung carcinoma cell), HCT116(human colon cancer cell) and A2780(Proliferation of Human Ovarian Cell) etc. various human tumor cell line, synthesized 7-virtue (first) amido-5-amido-6-7-azaindole derivatives dihydrochloride common manifestation goes out to be better than positive control (CYC202, IC 50=2.85 ~ 6.65 μMs) remarkable inhibiting activity (IC 50=0.00024 ~ 3.95 μMs), wherein particularly evident to the restraining effect of HCT116.
(2) wherein compounds Ⅳ d, IV e, IV f, IV g, IV hand IV jextracorporeal anti-tumor particularly outstanding, it is to the IC of above-mentioned cell 50value respectively 0.0038-0.16 μM, 0.0064-0.67 μM, 0.00076-0.24 μM, between 0.00024-0.043 μM, 0.0054-0.17 μM and 0.00077-0.023 μM.
The measuring method of embodiment 38, CDK1/Cyclin B kinase inhibiting activity:
Experiment material: CDK1 kinase activity detection kit (MBL, code 5235), demecolcine solution (sigma), Hela cell (Shanghai cell bank)
Experimental technique:
(1) CDK1 kinases extracts: when cell density reaches the degree of converging of 80%, add demecolcine solution 20ng/ml(final concentration), collect with cell scraper after 14 hours, add protein lysate, liquid nitrogen multigelation 6 times, stores, or is directly used in experiment under subzero 80 centigradetemperature conditions.
(2) Setup Experiments CDK1 kinase control group; Blank group; Administration group (first with DMSO, storing solution is diluted to 500 μMs, is then diluted to 100 μMs with DDW).After each sample is ready to by table 2, hatch 20min for 37 DEG C.
Note: kinase reaction damping fluid composition (1mM ATP, 10 × cdc2 reaction solution).
(3) in every pipe, add each 5ul of kinase substrate Biotinylatel MV peptide, suspendible gently, hatches 30min at 37 DEG C.
(4) often pipe adds phosphorylation reaction stop buffer 200ul.
(5) centrifugal 15s, 8 × 103rpm.
(6) get supernatant mixture 100ul to add and be coated with in the microwell plate of monoclonal antibody 4A4, incubated at room 60min.
(7) discard supernatant in pipe, add dcq buffer liquid, after patting gently, remove liquid, as method washs 5 times.
(8) in every hole, the POD liquid of 100uL is added, incubated at room 30min.
(9) operation of repeating step 7.
(10) every hole adds test kit substrate reagent, under room temperature and lucifuge condition, hatch 3-5min.
(11) in every hole, reaction terminating liquid 100ul is added.
(12) measure absorption value with spectrophotometer in wavelength 492nm place, the data obtained is for calculating inhibiting rate value.
Acquired results is see table 3, and table 3 is external Inhibiting enzyme activities of CDK1/Cyclin B (10 μMs) of part 7-virtue (first) amido-5-amido-6-7-azaindole derivatives dihydrochloride.
As can be seen from table 3 press down enzyme test result, under 10 μMs of concentration, 7-virtue (first) amido-5-amido-6-7-azaindole derivatives dihydrochloride synthesized by the overwhelming majority shows and is better than positive control cYC202significant CDK1 inhibit activities (inhibiting rate: 95.8% ~ 100%); Wherein part of compounds (IV a, IV c, IV d, IV e, IV i, IV j) 100%(10 μM is reached to the inhibiting rate of CDK1).
In sum, this compounds has good antitumor application prospect, thus has further developing drugs and is worth.

Claims (6)

1. 7-virtue (first) amido-5-amido-6-7-azaindole derivatives, it is characterized in that, general structure is:
Wherein:
Ar be without replacing, mono-substituted phenyl ring or fragrant heterocycle, the substituting group wherein on monosubstituted phenyl ring is fluorine, chlorine, bromine, nitro, alkoxyl group containing the straight or branched of 1 to 3 carbon atom; Virtue heterocycle is pyridine, pyrimidine;
N is 0 or 1;
R 1with R 2identical or different, select hydrogen, containing the alkyl of the straight or branched of 1 to 3 carbon atom and hydroxyalkyl; Or NR 1r 2for six-ring, this six-ring selects piperidines, morpholine or piperazine.
2. 7-virtue (first) amido-5-amido-6-7-azaindole derivatives, is selected from following compound:
T-butylbenzyl (6-piperidines-3-nitropyridine-2-base) carbamate,
T-butylbenzyl (6-morpholine-3-nitropyridine-2-base) carbamate,
T-butylbenzyl (6-diethylin-3-nitropyridine-2-base) carbamate,
T-butylbenzyl (6-(2-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate,
T-butylbenzyl (6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate,
T-butylbenzyl (6-(2-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate,
T-butylbenzyl (6-(4-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-3-nitropyridine-2-base) carbamate,
Butyl benzyl (6-((2-tertbutyloxycarbonyl oxygen base) ethyl-(N-tertbutyloxycarbonyl) amido)-3-nitropyridine-2-base) carbamate,
Butyl benzyl (6-((2-tertbutyloxycarbonyl oxygen base) ethyl-(N-tertbutyloxycarbonyl) amido)-3-nitropyridine-2-base) carbamate,
Butyl benzyl (6-bis-(2-tertbutyloxycarbonyl oxygen ethylamine)-3-nitropyridine-2-base) carbamate,
T-butylbenzyl 6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base (pyridine-2-methyl) carbamate,
T-butylbenzyl 6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base (pyridine-3-methyl) carbamate,
T-butylbenzyl 6-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-3-nitropyridine-2-base (pyridine-4-methyl) carbamate,
Butyl benzyl (5-(piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-morpholine-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-diethylamide-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-(2-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-(4-(2-tertbutyloxycarbonyl oxygen ethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-(2-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-(4-(2-tertiary butyloxycarbonyl yloxymethyl) piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-((N-tertbutyloxycarbonyl)-2-tertbutyloxycarbonyl oxygen ethylamine)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
Butyl benzyl (5-(1-tertbutyloxycarbonyl oxygen butane-2-(N-tertbutyloxycarbonyl) is amino)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate
Butyl benzyl (5-bis-(2-tertbutyloxycarbonyl oxygen ethylamine)-1H-pyrrolo-[2,3-c] pyridin-7-yl) carbamate,
N-benzyl-5-(piperidin-1-yl)-1H-pyrrolo-[2,3-c] pyridine-7-amine hydrochlorate,
N-benzyl-5-morpholine-1H-pyrrolo-[2,3-c] pyridine-7-amine hydrochlorate,
N 7-benzyl-N 5, N 5-diethyl-1H-pyrrolo-[2,3-c] pyridine-5,7-diamine hydrochloride,
2-(1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-2-yl) ethylate hydrochlorate,
2-(1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate,
(1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-2-yl) methylate hydrochlorate,
(1-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) methylate hydrochlorate,
2-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base amine) ethylate hydrochlorate,
2-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base amine) Kauri-butanol hydrochloric salt,
2,2 '-(7-benzyl ammonia-1H-pyrrolo-[2,3-c] pyridine-5-base amine) diethylate hydrochlorate,
2-(1-(7-(pyridine-2-ylmethyl ammonia)-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate,
2-(1-(7-(pyridin-3-yl methyl ammonia)-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate,
2-(1-(7-(pyridin-4-yl methyl ammonia)-1H-pyrrolo-[2,3-c] pyridine-5-base) piperidin-4-yl) ethylate hydrochlorate.
3. a preparation method for 7-virtue (first) amido-5-amido-6-7-azaindole derivatives and pharmaceutical salts thereof, be is characterized in that, realized by following steps:
(1) in polar solvent, under alkaline condition and 50-80 DEG C of temperature of reaction, there is aminated reacting generating compound II in chemical compounds I and corresponding aminated compounds, products therefrom can obtain sterling through column chromatography, polar solvent is N, dinethylformamide, methyl alcohol, ethanol, Virahol and methyl-sulphoxide, alkaline matter can select the one in salt of wormwood, sodium carbonate or sodium bicarbonate;
(2) in the anhydrous tetrahydrofuran solution of compound ii, the tetrahydrofuran solution of vinyl magnesium bromide is dripped under nitrogen protection ,-78 DEG C of temperature of reaction, and under-20 DEG C of conditions, react 16-24 hour generation compound III, the ammonium chloride solution adding 20% after reaction terminates in the solution carries out cancellation, and products therefrom can obtain sterling through column chromatography;
(3) compound III removes Boc protecting group, reaction times 12-24 hour in the saturated ethyl acetate solution of hydrochloric acid, generates the dihydrochloride of compounds Ⅳ, can obtain target compound sterling through suction filtration;
Synthetic route:
Wherein Ar, n, R 1and R 2definition with claim 1.
4. a kind of 7-virtue (first) amido-5-amido-6-7-azaindole derivatives according to claim 1 is preparing the application in antitumor drug.
5. a kind of 7-virtue (first) amido-5-amido-6-7-azaindole derivatives of preparing of method and pharmaceutical salts thereof are preparing the application in antitumor drug according to claim 3.
6. apply according to claim 5, it is characterized in that, described pharmaceutical salts selects hydrochloride, vitriol, phosphoric acid salt, tartrate, mesylate, benzene sulfonate, tosilate, Citrate trianion, maleate, fumarate and oxalate.
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1114506A (en) * 1993-09-10 1996-01-03 卫材株式会社 Bicyclic heterocyclic sulfonamide and sulfonic ester derivatives
CN101001859A (en) * 2004-06-09 2007-07-18 葛兰素集团有限公司 Pyrrolopyridine derivatives
CN101326187A (en) * 2005-12-15 2008-12-17 霍夫曼-拉罗奇有限公司 Pyrrolo[2,3-c]pyridine derivatives
CN101460497A (en) * 2006-04-04 2009-06-17 菲布罗根有限公司 Pyrrolo- and thiazolo-pyridine compounds as HIF modulators
US20100222352A1 (en) * 2005-09-23 2010-09-02 Yale University Compounds and Methods for the Treatment of Viruses and Cancer
CN101018789B (en) * 2004-09-03 2010-09-08 株式会社柳韩洋行 Pyrrolo[2,3-c]pyridine derivatives and processes for the preparation thereof
WO2011019634A2 (en) * 2009-08-10 2011-02-17 Taipei Medical University Aryl substituted sulfonamide compounds and their use as anticancer agents

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1114506A (en) * 1993-09-10 1996-01-03 卫材株式会社 Bicyclic heterocyclic sulfonamide and sulfonic ester derivatives
CN101001859A (en) * 2004-06-09 2007-07-18 葛兰素集团有限公司 Pyrrolopyridine derivatives
CN101018789B (en) * 2004-09-03 2010-09-08 株式会社柳韩洋行 Pyrrolo[2,3-c]pyridine derivatives and processes for the preparation thereof
US20100222352A1 (en) * 2005-09-23 2010-09-02 Yale University Compounds and Methods for the Treatment of Viruses and Cancer
CN101326187A (en) * 2005-12-15 2008-12-17 霍夫曼-拉罗奇有限公司 Pyrrolo[2,3-c]pyridine derivatives
CN101460497A (en) * 2006-04-04 2009-06-17 菲布罗根有限公司 Pyrrolo- and thiazolo-pyridine compounds as HIF modulators
WO2011019634A2 (en) * 2009-08-10 2011-02-17 Taipei Medical University Aryl substituted sulfonamide compounds and their use as anticancer agents

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Discovery of 1H-pyrrolo[2,3-c]pyridine-7-carboxamides as novel, allosteric mGluR5 antagonists;Manuel Koller 等;《Bioorganic & Medicinal Chemistry Letters》;20120821;第22卷(第20期);第6454-6459页 *
FEP-Guided Selection of Bicyclic Heterocycles in Lead Optimization for Non-Nucleoside Inhibitors of HIV-1 Reverse Transcriptase;Joseph T. Kim 等;《J. AM. CHEM. SOC.》;20061110;第128卷(第48期);第15372-15373页 *
Terry Panchal 等.Evaluation of basic, heterocyclic ring systems as templates for use as potassium competitive acid blockers (pCABs).《Bioorganic & Medicinal Chemistry Letters》.2009,第19卷(第23期),第6831-6817页. *

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