Summary of the invention
Technical problem to be solved by this invention is the deficiency that overcomes prior art, and a kind of new thiazole amide compound is provided, and it is the less protein tyrosine kinase inhibitor of a kind of spinoff.
For solving above technical problem, a kind of technical scheme that the present invention takes is following:
A kind of thiazole amide compound and pharmacologically acceptable salt thereof, it has the structure shown in the formula (I),
Wherein,
R
1Be hydrogen or deuterium;
R
2Be hydrogen, C
1-C
6Alkyl;
R
3Be hydrogen, the C of straight or branched
1-C
6Alkyl, perhaps R
3Representative-COO R
4, wherein, R
4C for straight or branched
1-C
6Alkyl.
According to an aspect of the present invention, R
2For being selected from-CH
3,-CH
2CH
3,-CH
2CH
2CH
3,-CH (CH
3) CH
3,-CH
2CH (CH
3)
2,-(CH
2)
3CH
3,-(CH
2)
4CH
3,-(CH
2)
5CH
3,-(CH
2)
2CH (CH
3) CH
3,-CH
2C (CH
3)
3,-C (CH
3)
3And-CH
2CH
2C (CH
3)
3In a kind of.
According to another aspect of the invention, R
3Representative-COOR
4, R wherein
4For being selected from-CH
3,-CH
2CH
3,-CH
2CH
2CH
3,-CH (CH
3) CH
3,-CH
2CH (CH
3)
2,-(CH
2)
3CH
3,-(CH
2)
4CH
3,-(CH
2)
5CH
3,-(CH
2)
2CH (CH
3) CH
3,-CH
2C (CH
3)
3,-C (CH
3)
3And-CH
2CH
2C (CH
3)
3In a kind of.
According to an aspect of the present invention, it provides formula (Ia) or the represented compound of formula (Ib).
The another technical scheme that the present invention takes is: thiazole amide compound and the pharmacologically acceptable salt thereof shown in a kind of formula (II),
Wherein,
R
1Be hydrogen or deuterium;
R
2Be hydrogen, C
1-C
6Alkyl;
R
3Be hydrogen, the C of straight or branched
1-C
6Alkyl, perhaps R
3Representative-COO R
4, wherein, R
4C for straight or branched
1-C
6Alkyl.
According to one aspect of the invention, R
2For being selected from-CH
3,-CH
2CH
3,-CH
2CH
2CH
3,-CH (CH
3) CH
3,-CH
2CH (CH
3)
2,-(CH
2)
3CH
3,-(CH
2)
4CH
3,-(CH
2)
5CH
3,-(CH
2)
2CH (CH
3) CH
3,-CH
2C (CH
3)
3,-C (CH
3)
3And-CH
2CH
2C (CH
3)
3In a kind of.
According to another aspect of the invention, R
3Representative-COOR
4, R wherein
4For being selected from-CH
3,-CH
2CH
3,-CH
2CH
2CH
3,-CH (CH
3) CH
3,-CH
2CH (CH
3)
2,-(CH
2)
3CH
3,-(CH
2)
4CH
3,-(CH
2)
5CH
3,-(CH
2)
2CH (CH
3) CH
3,-CH
2C (CH
3)
3,-C (CH
3)
3And-CH
2CH
2C (CH
3)
3In a kind of.
According to an aspect of the present invention, it provides suc as formula (IIa) or (IIb) represented compound.
Should be understood that the present invention's " compound " comprises any kind or all in this type form.
According to the present invention, described pharmacologically acceptable salt includes but not limited to hydrochloride, phosphoric acid salt, vitriol, acetate, PHENRAMINE MALEATE, metilsulfate, benzene sulfonate, toluenesulfonate, fumarate, tartrate etc.
Compound of the present invention can be used for preparing anti-malignant tumor medicine, and malignant tumour described here includes but not limited to chronic lymphocytic leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, myelodysplastic syndrome, lung cancer, ovarian cancer, prostate cancer, soft tissue sarcoma and glioblastoma etc.
The preparation of The compounds of this invention can be through the route of synthesis of well-known those the similar methods of chemical field, the synthetic compound of the present invention of description that particularly comprises according to this paper.Reagent generally obtains or is easy to use the well-known method preparation of those skilled in the art from commercial source.U.S. Patent number 6,596, the description of the preparation of 746 pairs of Dasatinibs will be incorporated into this paper as a reference.
Because the enforcement of above technical scheme, the present invention compared with prior art has following advantage:
1, the first pass effect of The compounds of this invention is very little, and bioavailability is very high;
2, The compounds of this invention has very high water solubility, can reach 15-20mg/mL, thereby is convenient to prepare its preparation and improves its effective bioavailability.
Embodiment
Hereinafter, the compound that provides has provided title and structural formula simultaneously, and wherein compound is as the criterion with structural formula, and name is called reference.
The RM that hereinafter uses, if do not provide the preparation method especially, promptly showing can be directly through being purchased acquisition.
Below in conjunction with specific embodiment the present invention is done further detailed explanation, but the present invention is not limited to following examples.
Embodiment 1
Formula (Ia) compound, its chemical name are N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-(2-amino-3-methyl-butyl carbonyl)-2,2,3,3,5,5,6,6-eight deuteriums-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-5-thiazole carboxamides.
Embodiment 2
Formula (Ib) compound, its chemical name be N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[and 4-(2-(N-tertbutyloxycarbonyl) amino)-3-methyl-butyl carbonyl)-2,2; 3,3,5; 5,6,6-eight deuteriums-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-the 5-thiazole carboxamides.
Embodiment 3
Formula (IIa) compound, its chemical name are N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-(2-amino-3-methyl-butyl carbonyl)-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-5-thiazole carboxamides.
Embodiment 4
Formula (IIb) compound, its chemical name are N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-(2-(N-tertbutyloxycarbonyl-amino)-3-methyl-butyl carbonyl)-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-5-thiazole carboxamides.
Embodiment 5
Present embodiment provides the preparation method of the compound that a kind of following formula representes,
Wherein, R
2, R
4Definition is with the summary of the invention part.
This compound can prepare through the synthetic route that following equation is represented:
Below with R in the formula
2Be sec.-propyl, R
4For the compound of tertiary butyl representative also is that formula (Ib) compound is an example, specify its preparation process.
The preparation process of formula (Ib) compound comprises the steps:
(1), preparation 1-tertbutyloxycarbonyl-deuterium is for piperazine: at room temperature, while stirring with sodium hydroxide (0.78g, 19.5mmol) aqueous solution of 1mL slowly is added drop-wise to deuterium for piperazine hydrochloride (3.25g; 19.4mmol) in, reacted about 5 minutes, and then drip 250mL methyl alcohol; Stir after 30 minutes, be reflected at the cryosel bath and stirred 30 minutes for following-20 ℃, slowly drip the methanol solution (4.25g of tert-Butyl dicarbonate simultaneously; 19.5mmol) about 50mL, be placed on room temperature reaction 1 hour, concentrate; Add water and stir 15 minutes after-filtration; Filtrating is used dichloromethane extraction, and organic phase is used anhydrous sodium sulfate drying, and filtering and concentrating also obtains clear crystal in vacuum-drying and is 1-tertbutyloxycarbonyl-deuterium for piperazine.
(2), preparation N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-tertbutyloxycarbonyl-2,2,3,3; 5,5,6,6-eight deuteriums-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-the 5-thiazole carboxamides: while stirring with 1-tertbutyloxycarbonyl-deuterium for piperazine (2.1g; 10.8mmol) and N, and the N-diisopropylethylamine (2.8g, (3.6g is 9.1mmol) in the DMSO 99.8MIN. 100mL solution for-5 thiazole carboxamides 21.7mmol) slowly to be added drop-wise to N-(2-chloro-6-methyl-phenyl)-2-[(6-chloro-2-methyl-4-pyrimidyl) amino]; Be heated to 115 ℃, under agitation reacted 18 hours, cool to room temperature adds water has deposition to separate out, and at room temperature stirs filtration in 15 minutes; Obtain light brown powder with rinsed, after obtain pure light brown product with re-crystallizing in ethyl acetate again and be N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-tertbutyloxycarbonyl-2,2; 3,3,5; 5,6,6-eight deuteriums-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-the 5-thiazole carboxamides.
Compound to preparation has carried out hydrogen nuclear magnetic resonance
1H NMR (400MHz, d6-DMSO) and mass spectrometric measurement, the result is following:
1Absorption peak in the H NMR spectrogram: 11.50 (s, 1H), 9.87 (s, 1H), 8.22 (s, 1H), 7.40 (m, 1H), 7.27 (m, 2H), 6.01 (s, 1H), 2.40 (s, 3H), 2.44 (s, 3H) .1.42 (s, 9H); M/s: [MH]+: 552.
(3), (2-chloro-6-aminomethyl phenyl)-[[6-(2,2,3,3,5 for 2-for preparation N-; 5,6,6-eight deuteriums-1-piperazinyl)-2-methyl-4-pyrimidyl] amino]-the 5-thiazole carboxamides: at room temperature, (2.4g is in 100mL dichloromethane solution 4.4mmol) while stirring the 5mL trifluoroacetic acid slowly to be added drop-wise to step (2) gained compound; Reacted 2 hours, and spun off solvent and obtain light brown powder, clean with saturated sodium hydrogen carbonate solution and stirred 15 minutes, filtration promptly gets N-, and (2-chloro-6-aminomethyl phenyl)-[[6-(2 for 2-; 2,3,3,5; 5,6,6-eight deuteriums-1-piperazinyl)-2-methyl-4-pyrimidyl] amino]-5-thiazole carboxamides (1.95g, 99.2%).
Compound to preparation has carried out hydrogen nuclear magnetic resonance
1H NMR (400MHz, d6-DMSO) and mass spectrometric measurement, the result is following:
1Absorption peak in the H NMR spectrogram: 9.88 (s, 1H), 8.22 (s, 1H), 7.41 (d, 1H), 7.27 (m, 2H), 6.01 (s, 1H), 2.40 (s, 3H), 2.24 (s, 3H).
m/z:[MH]
+=452。
(4), preparation formula (Ib) compound: at room temperature, while stirring with 2-(1H-benzo trisazo-L-1-yl)-1,1,3; 3-tetramethyl-urea Tetrafluoroboric acid ester (TBTU) (0.8g, 2.5mmol) be added drop-wise to uncle's 2-fourth oxanamide base-3 Methylbutanoic acid (0.43g, N 2.0mmol) is among the dinethylformamide 10mL; Under same condition, stirred 30 minutes, the back slowly adds N under nitrogen protection, and the N-diisopropylethylamine (0.64g, 5.0mmol); Stirred under the same condition 30 minutes, postcooling to 0 ℃ slowly drips step (3) gained compound (0.75g, N 1.66mmol) to this reaction system; Dinethylformamide solution 5mL after dropwising, returns to stirring at room and reacted about 18 hours, after reaction finishes; To wherein adding water, use ethyl acetate extraction, with saturated sodium chloride solution washing, add anhydrous sodium sulfate drying again; Filtering and concentrating vacuum-drying gets thick product, is formula (Ib) compound after 1: 19 mistake of methyl alcohol and methylene dichloride column purification obtains white powder (0.54g, 50.0%).
Compound to preparation has carried out hydrogen nuclear magnetic resonance
1H NMR (400MHz, d6-DMSO) and mass spectrometric measurement, the result is following:
1Absorption peak in the H NMR spectrogram: 11.64 (s, 1H), 9.89 (s, 1H), 8.22 (s, 1H), 7.40 (d, J=8.8Hz; 1H), 7.27 (m, 2H), 6.89 (d, J=8.8Hz, 1H), 6.06 (s; 1H), 4.27 (m, 1H), 2.43 (s, 3H), 2.24 (s, 3H); 1.99 (m, 1H), 1.38 (s, 9H), 6.86 (d, J=6.4Hz, 6H).
m/z:[MH]
+=651。
Embodiment 6
Present embodiment provides the preparation method of the compound that a kind of following formula representes,
In the formula, R
2Definition is with embodiment 5, and the compound that the compound that this formula is represented can embodiment 5 preparations is a raw material, under the trifluoroacetic acid effect, reacts to obtain.With R in the formula
2The compound of representing for sec.-propyl is an example, and it is following that it specifically prepares process:
At room temperature; (35mg in 1mL dichloromethane solution 0.054mmol), reacted 1 hour while stirring the 0.1mL trifluoroacetic acid slowly to be added drop-wise to formula (Ib) compound; Spin off solvent and obtain light brown powder; Clean with saturated sodium hydrogen carbonate solution again and stirred 15 minutes, filter and promptly obtain target product (21mg, 70.9%).
Compound to preparation has carried out hydrogen nuclear magnetic resonance
1H NMR (400MHz, d6-DMSO) and mass spectrometric measurement, the result is following:
1Absorption peak in the H NMR spectrogram: 9.88 (s, 1H), 8.22 (s, 1H), 7.29 (d, J=6.0Hz, 1H), 7.25 (m, 2H), 6.07 (s, 1H), 2.43 (s, 3H), 2.24 (s, 3H), 1.75 (m, 1H), 0.90 (d, J=6.4Hz, 3H), 0.82 (d, J=6.8Hz, 3H).
m/z:[MH]
+=551。
Embodiment 7
Present embodiment provides a kind of N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-(2-(N-butyl ester base-amino)-3-methyl-butyl carbonyl)-2,2,3; 3; 5,5,6; 6-eight deuteriums-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-preparation method of 5-thiazole carboxamides mesylate (being the mesylate of formula (Ib) compound), specific as follows:
At room temperature; While stirring with methylsulfonic acid (18mg, (100mg is in 5mL methanol solution 0.184mmol) 0.187mmol) slowly to be added drop-wise to formula (Ib) compound; Reacted 1 hour, removing desolvates obtains the mesylate (yield 89%) that white powder 105mg is formula (Ib) compound.
Embodiment 8
Present embodiment provides a kind of N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-(2-(N-butyl ester base-amino)-3-methyl-butyl carbonyl)-2,2,3; 3; 5,5,6; 6-eight deuteriums-1-piperazinyl]-2-methyl-4-pyrimidyl] amino]-preparation method of 5-thiazole carboxamides mesylate (being the Citrate trianion of formula (Ib) compound), specific as follows:
At room temperature, while stirring with the Hydrocerol A (monohydrate) of 1ml (39mg, 0.185mmol) methanol solution slowly is added drop-wise to (100mg; 0.184mmol) the 5mL methanol solution in, be heated to 80C then, reacted 1 hour; Temperature drops to room temperature; Removing desolvates obtains white powder, and with the anhydrous diethyl ether washing, 118mg is Citrate trianion (yield 88.7%) again.
Embodiment 9
Present embodiment provides the preparation method of the compound that a kind of following formula representes,
Wherein, R
2, R
4Definition is with the summary of the invention part.
This preparation method comprises the steps:
(1), through the synthetic N-(2-chloro-6-deuterium is for methyl-phenyl) of following synthetic route-2-[(6-chloro-2-methyl-4-pyrimidyl) amino]-5 thiazole carboxamides
(2), be raw material with step (1) gained N-(2-chloro-6-deuterium is for methyl-phenyl)-2-[(6-chloro-2-methyl-4-pyrimidyl) amino]-5 thiazole carboxamides, through obtaining title product with the identical method of embodiment 5 steps (2) to step (4).
The test of pesticide effectiveness
One, tumour cell inhibition test:
1, TP
(1), compound: earlier test compounds is dissolved among the 100%DMSO in the in vitro study, redilution is to desired concn, and the final concentration of DMSO is 0.1%.The DMSO of 0.1% (v/v) is added substratum as solvent control, totally 9 concentration gradients, repeated test secondary.
(2), tumor cell line: the tumour cell of surveying ties up in RPMI1 0 substratum that contains 10% foetal calf serum, in 5%CO
2, cultivate in 37 ℃ of incubators.The tumor cell line of surveying is: 1) A375, melanoma (Melanoma) tumour cell; 2) A673, neck struma oncocyte; 3) HepG2, tumor cell of liver; 4) U87, neurospongioma (Glioma) tumour cell; 5) K562, white blood disease (Leukemia) tumour cell; 6) MDA-MB-231, the breast cancer tumour cell; 7) A549 and H460, the nonsmall-cell lung cancer tumour cell; 8) HT29, the colorectal carcinoma cell; 9) PC-3, the prostate cancer tumour cell; 10) Mia-PaCa-2, the pancreatic tumour cell.
(3), CellTiter-Glo cytoactive fluoroscopic examination test: cell inoculation in 96 orifice plates, 3000 cells in every hole, and at 5%CO
2, incubated overnight in 37 ℃ of humidification incubators.After adding test compounds in the hand-hole in second day, hatched again 72 hours.Use the CellTiter-Glo cytoactive fluorescence detection reagent kit of Promega company to detect cell activity.Calculate IC
50(compare to make the cell growth receive 50% suppress required drug level, use the nonlinear regression analysis of GraphPad Prism software to calculate) with the DMSO control group.
(4), sample analysis:
1) contains and add the detection reagent for preparing in 96 orifice plates of 100 μ L cell culture mediums and carry out viable cell and detect, do not have cell (only containing substratum) to contrast as a setting in the plate hole, have only substratum not have detection reagent as experiment contrast.Hatch according to culture scheme.
2) the about 30min of balance plank and test sample under the room temperature.
3) add equal-volume (100 μ L) CellTiter-Glo
TMReagent mixes 2min gently on the vortice, incubated at room 10min makes luminous signal stable.CellTiter-Glo
TMThe luciferase test kit is united and is used for viable count and quantitatively provides easily and fast a kind of and the sensitive method, realizes that through quantitative ATP it is the semiochemicals in the viable cell metabolism.CellTiter-Glo
TMThe viable cell detection kit adopts luciferase to do to detect thing, because there is not the interference of endogenous luciferase in the mammalian cell, and the steady glow type signal that uses the UltraGlow luciferase to generate in the test kit, the transformation period surpasses 4h.The luminous signal of overlength is that many plates provide the foundation with batch detection.Luciferase needs the participation of ATP in the luminescence process, has the respiration of metabolic activity cell and other vital movement processes can produce ATP.In cell culture medium, add equal-volume CellTiter-Glo
TMReagent is measured luminous value, and the ATP amount is directly proportional in optical signal and the system, and ATP and viable count positive correlation.
4) plate is put into the multi-functional luminosity meter of Modulus microwell plate, click " Start " and begin to detect, after detection finished, take off data can show that in detecting, run into other problems, the problem introduction of please refer to obtains more information with the Excel tabulated form.
5) detect to finish just can to carry out data analysis and calculate 50% inhibition concentration IC through Excel
50
2, test-results
See also table 1.
Table 1
Visible from table 1, each compound of test has all showed inhibition activity in various degree to various tumour cells, has particularly shown that for wild-type BCR-ABL (K562) cell strain growth significant inhibition is active.
Two, the restraining effect of human leukemia tumour cell transplanted tumor in nude mice test
1, TP: 20 of nude mices (male, 6 ages in week), inoculation K562 human leukemia tumour cell treats that the knurl average-volume reaches 300mm
3The time, be divided into 2 groups at random, be respectively control group and 10mg/kg/ days formula (Ib) compound dosage group, successive administration 10 days, vein or subcutaneous injection.Write down twice tumour size and body weight weekly from medication treatment beginning in first day.If medication caused>20% dead and/or 20% would lose weight only then think ' toxic '.With formula (l * w
2) calculate the weight of tumour, the wherein minimum and maximum size (mm) of the each measurement of l and w representative.Draw graph of a relation and the nude mice mean body weight that the fate of tumor average volume after with tumour transplatation change respectively according to result calculated and change the graph of a relation that changes with the fate after the tumour transplatation.
2, result: referring to Fig. 1 and Fig. 2; The test of human leukemia tumour cell transplanted tumor in nude mice shows that The compounds of this invention has extremely strong restraining effect to the white blood disease tumour, and the nude mice tumour promptly disappears or completely dissolve after with a course of treatment of The compounds of this invention treatment basically.Simultaneously, nude mice mean body weight result of variations shows that the general toxicity of The compounds of this invention is very little, and its resistance is very high.
Similar with Dasatinib, compound of the present invention can be used for but is not limited to treat chronic myelocytic leukemia (CML), and it can combine with dissimilar pharmaceutical salts and process oral prepns (tablet or capsule etc.).The tablet of processing with The compounds of this invention or capsule can be taken once a day or repeatedly.The compounds of this invention also can combine to process compound preparation with other its medicines.
The foregoing description only is explanation technical conceive of the present invention and characteristics, and its purpose is to let the personage who is familiar with this technology can understand content of the present invention and enforcement according to this, can not limit protection scope of the present invention with this.All equivalences of doing based on spirit of the present invention change or modify, and all should be encompassed within protection scope of the present invention.