CN103319476A

CN103319476A - Kinase inhibitor

Info

Publication number: CN103319476A
Application number: CN2013102332952A
Authority: CN
Inventors: 郝岩
Original assignee: JINAN DEAI PHARMACEUTICAL TECHNOLOGY Co Ltd
Current assignee: JINAN DEAI PHARMACEUTICAL TECHNOLOGY Co Ltd
Priority date: 2013-06-13
Filing date: 2013-06-13
Publication date: 2013-09-25
Anticipated expiration: 2033-06-13
Also published as: CN103319476B

Abstract

The invention relates to a kinase inhibitor, which is capable of inhibiting kinase, and particularly inhibiting the activity of BCR-ABL kinase. The inhibitor disclosed by the invention is of a structure shown in formula (I) which is defined in the specification, wherein the compound can be used for preparing a medicine composition, namely, used for kinase inhibition for human or animals; for example, the compound can be used for treatment of tumour and/or other diseases.

Description

A kind of kinase inhibitor

Technical field

This invention relates to the pharmaceutical chemistry field, is specifically related to a kind of compound, comprises the pharmaceutical composition of this compound, and as the purposes of kinase inhibitor.

Background technology

Kinases is a kind of enzyme that changes the proteolytic enzyme chemical structure by phosphorylated.Phosphorylated usually can make proteolytic enzyme the performance cell the position and change with the combination of other albumen.The function of 30% albumen is by kinases control, and kinase regulatory the passage of albumen, the signal transmission between the cell.Human body gene contains more than 500 kind of kinase gene, accounts for 2% of all genes.

Protein kinase is proteolytic enzyme relevant on one group of structure, and they can transfer to phosphoryl group the Serine of enzyme from nucleoside triphosphate, on Threonine and the tyrosine.Permitted cellulous function, such as dna replication dna, Growth of Cells, energy metabolism etc. is to control by phosphorylated by kinases all.It is pointed out that more numerous disease (comprising cancer) also regulated and control by protein kinase.Known more than 100 kinds of Common Cancer genes in, tenuigenin Tyrosylprotein kinase or suppressed, or activated widely.((Blume-Jensen?and?Hunter,Nature,411:355-365(2001))。Naturally, protein kinase has become the target of drug research, and existing more than 10 kind of kinase inhibitor is approved for medicine.(summary, current medicine, 11:1563 (2004); Dancey and Sausville, NatureReivews Drug Discovery, 2:296 (2003)).

The growth factor receptors that has protein-tyrosine to activate enzyme is again the acceptor junket ammonia activating enzyme factor.They have a variety of, are regulating and control a lot of cytoactives.They form by having in a large number different bioactive cell-membrane receptors.At least 19 kinds of these Class Activation enzymes are arranged, comprise VEGFR-1, VEGFR-2, Flt-3 and PDGFR.

Chronic myeloid leukemia (CML) is a Diseases of Hematopoietic Stem Cell, and it is bred in the hemocyte tissue and cause owing to containing the kinase whose cell of BCR-ABL (Breakpoint Cluster Region-Abelson).BCR-ABL oncogene is the product that the expense drawstring takes inferior karyomit(e) (Ph) 22q.Its decoding reaches a carat BCR-ABL protein makes it have the ABL tyrosine-kinase active, and this also is the cause of disease of chronic myeloid leukemia (CML).Although 210kDa BCR-ABL protein enzyme activates fully among the chronic myeloid leukemia people, the proteolytic enzyme of a 190kDa BCR-ABL also is activated among the leukemia people of demonstration Ph positive (Ph+).Gleevec is ABL albumen activation inhibitor, thereby the while is also suppressed growth factor receptors PDGFR and KIT reaches the anticancer growth.Its application has greatly changed cancer therapy.This targeted therapeutics not only provides effective therapy for chronic myeloid leukemia people, has also changed the history of cancer therapy.Some patient can develop immunity to drugs to Gleevec.This is because the BCR-ABL protein enzyme produces the structure change, thereby insensitive to Gleevec.Overcoming the drug-fast method of Gleevec has: select to suppress one-piece construction or the stability of BCR-ABL protein enzyme, suppress the signal transduction system of BCR-ABL protein enzyme.A lot of biochemical structure researchs make people resist the clearer of property of medicine understanding, thereby design new medicine to avoid the resistance of human body.Many novel compounds are having drug-fast chronic myeloid leukemia people to carry out first phase or the second stage of clinical experiment to Gleevec, two s-generation medicines demonstrate significant curative effect.

Modal patient's resistance is the point mutation owing to the BCR-ABL gene, thereby makes activating enzyme at one or several amino acid change of catalytic domain, and kinase inhibitor just cannot be followed the kinases combination like this, and cancer cells is grown in the same old way.

Deuterium (D or ²H) be the isotropic substance of a stable hydrogen, its atomic mass is 2.0144.Hydrogen exists with three kinds of isotropic substance forms at occurring in nature: ¹H (protium), D ( ²H or deuterium) and T ( ³H or tritium).The abundance of deuterium is 0.015%.If surpass 0.015% deuterium so contain in the compound, they just can not be that nature exists, and that is to say that compound is brand-new.

Dasatanib is s-generation BCR-ABL kinase inhibitor.U.S. Patent number 6,596746; U.S. Patent application US2005/0176965A1 and U.S. Patent application US2006/0176965A1.It is for Gleevec drug-fast patient to be arranged.But its transformation period is only less than four hours, and medicine is for studies show that it is decomposed much in vivo.（Christopher?et?al,Drug?Metablism?and?Disposition,36,7,1357-1364.)。

Summary of the invention

The invention provides a kind of compound, it contains deuterium for ethanoyl, can suppress kinase whose activity, particularly the BCR-ABL kinases.Compound of the present invention can be used for pharmaceutical compositions, namely is used for human or animal's kinase inhibition, for example is used for the treatment of tumour and/or other diseases.Therefore, the compounds of this invention can be treated malignant tumour (such as chronic myeloid leukemia, lung cancer, ovarian cancer, mammary cancer, carcinoma of the pancreas, bladder cancer, colorectal carcinoma is etc. the essence cancer).

Compound of the present invention has the structure of formula (I):

The invention still further relates to the pharmacy acceptable salt of above-claimed cpd.Suitable pharmacy acceptable salt comprises mineral acid and organic acid subsalt: example hydrochloric acid,, Hydrogen bromide, sulfuric acid, phosphoric acid, methylsulfonic acid, trifluoromethanesulfonic acid, Phenylsulfonic acid, tosic acid, 1-naphthalene sulfonic aicd, 2-naphthene sulfonic acid, acetic acid, lactic acid, trifluoroacetic acid, oxysuccinic acid, tartrate, citric acid, oxalic acid, fumaric acid, succsinic acid, toxilic acid, Whitfield's ointment, phenylformic acid, toluylic acid, amygdalic acid etc.; In addition, also comprise the acid salt of mineral alkali, as contain the salt of alkali metal cation, alkaline earth metal cation and ammonium cation, and the acid salt of organic bases, comprise by aliphatics and the ammonium of aromatic series replacement and the salt of quaternary ammonium cation.

The present invention also provides a kind of pharmaceutical composition, and it comprises compound or its pharmacy acceptable salt and the pharmaceutically acceptable carrier of formula (I).This pharmaceutical composition can be hard capsule, suspensoid, powder, granule, injection, non-aqueous liquid preparation or O/w emulsion.

The present invention also provides compound or the purposes of its pharmacy acceptable salt in the preparation kinase inhibitor of formula (I), and especially, described kinases is the BCR-ABL kinases.

The subordinate list explanation

Table 1 is the active testing data of the compound of formula (I)

Table 2 is active testing data of compound S taurosporine

Embodiment

The compounds of this invention can with known chemical reaction and process, be prepared from by commercially available chemical feedstocks.The preparation method provides specific embodiment in the experiment embodiment implementation section.

The compounds of this invention can give or with the administration of unit formulation formulation by oral, local, injection, suction, spraying or rectum, per os, skin, parenteral." drug administration by injection " comprises intravenous injection, intramuscular injection, and subcutaneous injection and parenteral injection, and use infusion techn.

The compounds of this invention can be prepared into pharmaceutical composition according to suitable pharmaceutical methods known in the art and pharmaceutically acceptable carrier.Can contain the auxiliary material of one or more screenings in the aforementioned pharmaceutical compositions, comprise thinner, sweeting agent, seasonings, tinting material and sanitas.Contain activeconstituents in the tablet, they mix with pharmacy non-toxic excipients approval, that be suitable for tablet manufacturing.Described vehicle is inert diluent, such as calcium carbonate, yellow soda ash, lactose, calcium phosphate or sodium phosphate; Granulating agent and disintegrating agent (such as W-Gum or alginic acid), tamanori (such as Magnesium Stearate, stearic acid or talcum powder).Tablet can also can carry out dressing by known technology without dressing, to postpone its disintegration and absorption in gi tract, in order to long-term lasting drug effect is provided.For example, can adopt such as slow-release materials such as glyceryl monostearate or distearins.These compounds also can be made fast release solid formulation.

Pharmaceutical composition of the present invention can be made different dosage form, such as hard capsule, and suspensoid, powder, granule, non-aqueous liquid preparation and O/w emulsion.

Should be noted that, the concrete dosage level of specific client need is had nothing in common with each other, depend on many factors, comprise the severity of activity, patient age, body weight, healthy state, sex, food habits, daily schedule, the mental status, the medicine velocity of discharge, drug regimen and the disease for the treatment of of used particular compound.

The compounds of this invention can be by general preparation method's preparation of known compound, and following examples as an illustration.

Embodiment

Except as otherwise noted, all reactions all in the glassware of flame drying or oven drying, are carried out with magnetic agitation under the drying nitrogen environment.Sensitive liquid and solution add reaction vessel with injection or conduit by the rubber peel plug.

All temperature of report are uncorrected degree centigrade (° C).Except other have indicate, all shares and per-cent are all calculated by weight.

The commercial reagent and the solvent that use do not carry out secondarily purified.

Use prefabricated Whatman silica gel 60A GF254 thin layer of glass plate (250 μ m) to carry out thin-layer chromatography (TLC).Thin layer plate is inspected and can be adopted following one or several technology: 1) uviolizing, 2) put in the iodine vapor 3) spray is with 10% phospho-molybdic acid ethanol, heating develops the color, and 4) spray with cerous sulfate solution the heating colour developing.Column chromatography uses 230-400 purpose EM Science silica gel G.

Fusing point (mp) measure to use the melting point apparatus of Thomas-Hoover(thomas-Hu Fo).Proton (1H) nucleus magnetic resonance (NMR) spectrum, can adopt Varian400(400HZ) nuclear magnetic resonance analyser, with Me4Si (δ 0.00ppm) or remaining protonic solvent (CHCl3, δ 7.26ppm, MeOH δ 3.30ppm, DMSO δ 2.49ppm) detects for standard.Carbon (13C) nucleus magnetic resonance (NMR) spectrum can adopt Varian400 (400Hz) nuclear magnetic resonance analyser, and (CDCl3 δ 77.0, MeOD δ 49.0, DMSO δ 39.5) detects as standard with solvent.Can obtain Algorithm (MS) and high resolution mass spec (HRMS) with electron impact (EI) or fast atom bombardment MS (FAB).

The structure of all compounds is all passed through nuclear magnetic resonance spectrum (NMR), and mass spectrum (MS) is proved conclusively.

Embodiment 1: the preparation of formula (I) compound

The synthetic route of the compound of formula (I) (being the compound (4) in the following reaction) is as follows:

By said synthesis route as can be known, the preparation of formula (I) compound is divided into following three steps.

The preparation of compound (2):

N-Boc-N '-methyl (D ₃) piperazine (2): compound 1 (1.0g, 5.37mmol) and K ₂CO ₃(2.23g, 3.22mmol) is dissolved in dry tetrahydrofuran (THF) (10mL), stirring at room 1h.Be cooled to-12 ° of C, drip deuterium for tetrahydrofuran (THF) (4mL) solution of methyl iodide (1.01g, 6.98mmol), then stirred overnight at room temperature.Reaction solution is concentrated, is dissolved in ethyl acetate (50ml), uses 1N NaOH(20ml) wash.Product is crossed column purification

（DCM/MeOH=50/1，560mg,51%)。 ¹H?NMR(300MHz,CDCl ₃),1.44(s,9H),2.32(t,4H),3.41(t,4H).

The preparation of compound (3):

N-methyl (D ₃) piperazine (3): ice bath in methylene dichloride (3mL) solution of compound 2c (490mg, 15.62mmol), drips trifluoroacetic acid (2mL).Stirring at room 3h, vacuum desolvation, crude product is directly used in next step.

The preparation of compound (4) (being the compound of formula (I)):

N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-methyl (D3)-piperazine-1-yl]-2-methylpyrimidine-4-yl] amino]-1,3-thiazole-5-methane amide (4c): add N-(2-chloro-6-aminomethyl phenyl)-2-[(6-chloro-2-methyl-4-pyrimidyl in the reaction flask) amino]-5-thiazole carboxamides (170mg, 0.43mmol), compound 3 (248mg, 2.41mmol), N, dinethylformamide (3mL) and diisopropyl level ethylamine (0.45g, 3.45mmol), regulate PH9-10.110 ° of C reactions are spent the night.Add entry (5X) at 110 ° of C, separate out solid, reaction solution is cool to room temperature slowly, stirs 2h, filters and obtains product.Product washes with water, vacuum-drying (178mg, 90%). ¹HNMR(300MHz,DMSO-d6),δ:8.22(s,1H),7.38(d,1H,J=7.0),7.23-7.29(m,2H),6.05(s,1H),3.51(m,4H),2.40(s,3H),2.38(m,4H),2.23(s,3H).M.p.296-299°C.

Embodiment 2: the kinase inhibiting activity of formula (I) compound is measured

The test of the kinase activity of formula (I) compound is by Reaction Biology Corp.(One Great Valley Parkway Suite2, Malvern, PA19355) the scientific research personnel finish.As object of reference, concrete experimental procedure is as follows with Staurosporine:

Reaction buffer:

Alkali reaction buffer reagent: 20mM Hepes (pH7.5), 10mM MgCl ₂, 1mM EGTA, 0.02%Brij35,0.02mg/ml BSA, 0.1mM Na ₃VO ₄, 2mM DTT, 1%DMSO

Reactions steps:

1. reaction substrate is joined in the freshly prepd alkali reaction buffer reagent.

2. add needed other subsidiary-phosphatide and calcium ion.

3. join the kinases of appointment in the mixture and shake up.

4. adding the compound (compound 4 or Staurosporine) that needs to measure activity reacts in mixture.

5. add ³³P-ATP (final activity be 0.01 μ Ci/ μ l) activating reaction in the reaction.

6. reaction was at room temperature preserved 120 minutes.

7. reaction solution is put (Whatman#3698-915) on the P81 ion exchange paper.

8. wash with 0.75% phosphoric acid liquid.

Experimental result

Referring to table 1 and 2.

Table 1, the activity of formula (I) compound

The concentration (M) of adding formula (I) compound	Active (%)
		1.00E-05	1.48
3.33E-06	1.52
		1.11E-06	1.46
3.70E-07	1.26
		1.23E-07	1.20
4.12E-08	2
		1.37E-08	2.02
4.57E-09	2.02
		1.52E-09	2.04
5.08E-10	3.45
		0.00E+00	100

Table 2, the activity of Staurosporine

Can find out that according to experimental result formula (I) compound has good inhibition activity to the BCR-ABL kinases.Known compound Staurosporine in test process, present IC50 (503nhibiting concentration) be 58.8 nmoles/liter.The compound of formula (I) in test process, present IC50 (503nhibiting concentration) for be far smaller than 0.5 nmole/liter, be 0.0063 nmole/liter.In accordance with international practices, 0.5 nmole/when rising, compound may be defined as has extremely highly active compound in order to be far smaller than as compound I C50 (503nhibiting concentration).

Claims

1. the compound of formula (I)

Or its pharmacy acceptable salt.

2. the compound of claim 1 or its pharmacy acceptable salt, described pharmacy acceptable salt comprises the subsalt of mineral acid, organic acid subsalt, the acid salt of mineral alkali, or the acid salt of organic bases.

3. pharmaceutical composition comprises compound or its pharmacy acceptable salt and the pharmaceutically acceptable carrier of claim 1.

4. the pharmaceutical composition of claim 2, described pharmaceutical composition is hard capsule, suspensoid, powder, granule, injection, non-aqueous liquid preparation or O/w emulsion.

5. the compound of claim 1 or its pharmacy acceptable salt are in the purposes of preparation in the kinase inhibitor.

6. purposes claimed in claim 4, wherein said kinases is the BCR-ABL kinases.

7. the synthetic method of the compound of claim 1 comprises the steps:

(1) N-Boc-N '-methyl (D ₃) preparation of piperazine

(2) N-methyl (D ₃) preparation of piperazine

(3) N-(2-chloro-6-aminomethyl phenyl)-2-[[6-[4-methyl (D3)-piperazine-1-yl]-2-methylpyrimidine-4-yl] amino]-preparation of 1,3-thiazoles-5-methane amide