CN102786469B - Adjacent pyridine hydrazide derivatives and preparation method thereof and pharmaceutical composition and purposes - Google Patents

Abstract

The present invention relates to 1 (4 chlorine 3 trifluoromethyl) 3 (4 (2 (2 isopropylidene hydrazine carbonyl) the pyridine 4 epoxide) phenyl) urea shown in Formulas I and II and 1 (4 chlorine 3 trifluoromethyl) 3 (4 (2 (2 isopropyl hydrazine carbonyl) pyridine 4 epoxide) phenyl) urea, its officinal salt, and preparation method thereof, containing the compositions of these compounds one or more, purposes disease such as immune disorder and tumor disease in terms of relevant with protein kinase in treatment with this compounds.

Description

Adjacent pyridine hydrazide derivatives and preparation method thereof and pharmaceutical composition and purposes

Invention field

The present invention relates to 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(the 2-isopropylidene hydrazine carbonyl shown in Formulas I and II Base) pyridine-4-epoxide) phenyl) urea and 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropyl hydrazine carbonyl) pyridine- 4-epoxide) phenyl) urea, its officinal salt, and preparation method thereof, containing the compositions of these compounds one or more, and such Purposes in terms of disease such as immune disorder and tumor disease that compound is relevant with protein kinase in treatment.

Background of invention

Recent years, due to the raising of the understanding of the biomolecule relevant to disease with some other to enzyme, greatly promote Having entered discovery or the development of the new drug for the treatment of disease, protein kinase is exactly an a kind of widely studied important class, and it is one Extended familys, the control with intracellular various signal transduction processes is relevant.Due to their structure and catalysis conservative it Be considered to evolve from common ancestral gene.Nearly all kinases all contains a 250-300 similar ammonia Base acid catalysis territory.These protein kinases are divided into multiple family according to the difference of phosphorylated substrate, such as protein tyrosine kinase, egg White serine/threonine kinase, lipoid etc..Typically, protein kinase turns from a ribonucleoside triphosphote by affecting a phosphoryl Move on to a protein receptor relevant to signal transduction pathway and carry out signal transduction in mediated cell.These phosphorylated events are as dividing The biological function of sub-switch-mode regulation target protein, is finally excited and reacts various extracellulars and other stimulation.Kinases exists In multilayer signal transduction path, receptor tyrosine kinase is positioned at the upstream of tumor-blood-vessel growth Signal transduction pathway and tumor is thin The upstream of born of the same parents' Signal transduction pathway.Serine/threonine protein kitase is positioned at the signal of tumor and tumor-blood-vessel growth cell and turns The downstream of guiding path.Research shows, by upstream retardance VEGFR and pdgf receptor, to block Raf/MEK/ERK in downstream, it is possible to Reduce the angiogenesis of tumor simultaneously and suppress the duplication of tumor cell, thus hindering the growth of tumor.

Raf kinases is the protein product encoded by proto-oncogene raf, is made up of 648 aminoacid, and molecular weight is 70000 ～74000D, containing 3 conserved region in its structure, respectively CR1 (61～194D), CR2 (254～269D), CR3 (335～ 627D).CR1 is positioned at its molecule possessing amino, rich in cysteine, containing zinc-finger structure, with the ligand binding domain of Protein kinase C Structure is similar, is the main portions that is combined of Ras with Raf-1 protein kinase of activation.CR2 is also near aminoterminal, rich in serine And threonine.CR3 is positioned at the c-terminus of its molecule, is the catalysis district of protein kinase.As Ras/Raf/MEK/ERK path In a Key kinases, Raf can be by relying on or play by the way of being independent of Ras the conduction regulation effect of its signal.As Raf Kinase whose stream substrates, the MEK phosphorylated CREB of activation, regulate various cell function.Once this path generation excessive activation, then Cause cell proliferation to accelerate with the cells survival phase to extend, thus lead oncogenic generation.

Research shows, the oncogene of more than 80% and proto-oncogene are present in the cancer encoding proteins tyrosine kinase of people (PTK), in, the generation of the various cancer of the mankind and development are to conduct with the abnormal cell signal coming from protein tyrosine kinase to have Close, an increase being mainly characterized by tyrosine kinase activity of malignant cell.Therefore, suppression tyrosine kinase activation or Block its signal conducting path and become the new way controlling tumor.

Endothelial growth factor receptor (EGFR) is a kind of protein tyrosine kinase receptor (RTK), is positioned at No. 7 chromosome P13～q22 district, total length 200kb, it is made up of 28 exons, encodes 1186 aminoacid, its glycoprotein molecule amount is about 170kDa, is distributed widely in all histiocytes in addition to ripe Skeletal Muscle Cell, parietal endoderm and hemopoietic tissue.EGFR There are the acceptor molecule that 4 structures are similar: ErbB1 (EGFR), ErbB2 (HER2), ErbB3 (HER3), ErbtM (HER4) in family, Belong to receptor tyrosine kinase (RTKS).They all contain the outer ligand binding domains of 1 born of the same parents, 1 membrane spaning domain and 1 There is the cytoplasmic domain of tyrosine kinase activity.Its intracellular region and erbB oncoprotein very high homology.The activation of EGFR Can be realized by the Receptor dimerization effect that part is induced.In ErbB receptor family, in addition to HER2, other members have it Respective ligand, various parts are to be come through Proteolytic enzyme by corresponding transmembrane protein precursor, have 1 EGF sample Domain.The part specific binding with EGFR includes epidermal growth factor (EGF), transforming growth factor α (TGF-α), two-way tune Joint albumen (AR), beta cell element (BTC), Heparin-binding EGF like growth factor (HB-EGF), epiregulin (EPR) etc..Outside born of the same parents Cause ErbB2 configuration to change after ligands, EGF (endothelial cell growth factor (ECGF)) and ErbB2 are specific binding, cause Receptor dimerization thus Activate their cytoplasmic location.After the intracellular region tyrosine phosphorylation of ErbB2 so activation second message,second messenger transduction, pass through MAPK The activation (regulation kinases Erk1 and Er1) of (mitogen protein kinase) approach inducing cell external signal: by PDK (phosphatidyl-4 Alcohol kinases) pathway activation signal transducer JAK；Start the transcription activators of STAT1, STATS3 further；On the other hand, cell Interior signal is by the ERK (extracellular regulated protein kinase), Jin Erjie in 6rb2 (growth factor receptor binding protein precursor) activation downstream Lead the transcription activating of ATF, NF-kB, Ap-1, C-fos and C-Jun.These are all the growths or carcinogenic that EGFR is mediated Basic downstream pathway.Abnormal EGFR activation mechanism include the amplification of receptor itself, the process LAN of receptors ligand, Activating mutations with And the shortage of negativity regulation approach, therefore EGFR induced cancer is at least through 3 kinds of mechanism: the process LAN of EGFR part, EGFR's The sudden change activation of amplification or EGFR.In these 3 kinds of mechanism, the sudden change activation of EGFR is to cause tumor cell aberrant biological behavior Main factor.Some sudden change of EGFR gene can cause receptor effect to strengthen and the prolongation of persistent period.Lynch etc. prove Variation receptor has no effect on the stability of receptor protein, is found by Tyr1068 phosphorylation assay EGFR activation, and wild type is subject to The activation 15min of body i.e. lowers, and the receptor that makes a variation shows the effect of higher than normal EGFR 2 times, and the continuous activation more than 3h.

EGFR sudden change does not affects the ability that tumor cell combines with TKI (tyrosine kinase inhibitor).TKI is to those The reason causing EGFR to activate because of sudden change can be explained by oncogeneaddiction model.Pass through Ras.Raf- MEK.ERK1/ERK2, PI3K.Akt, STAT3/STAT5 path, EGFR discontinuity height activates downstream signal, starts EGFR regulation Anti-apoptotic and survival signaling, cause cancer cell to become dependent upon this signal to maintain it to survive--i.e. have oncogene (sudden change The feature that EG relies on；After using specificity T KI to block EGFR signal, its proliferative effects and output survival signaling will be eliminated, Cause death of neoplastic cells.Result, it is believed that the variation of signal transduction pathway is the basis that medicine height is sensitive occur in cancer cell. On the contrary, the tumor cell (reactionless to Gefitinib, Erlotinib) that normal cell or non-EGFR rely on is unaffected.Because Existence is also ordered about by other genes, or can be made up by other RTK after EGFR suppresses.In oncogene relies on model, The oncogene that cell carcinomas relies on can produce apoptosis and the output of 2 signals of existence simultaneously.Under general Sui condition, oncogene is swashed Live.Survival signaling is occupied an leading position, and apoptotic signal is in low relative levels, makes cancer cell maintain growth and propagation.Work as cancer After the acute inactivation of gene, at crucial window phase, first it is that existence significantly weakens rapidly.And apoptotic signal slowly declines.Cause This causes signal uneven (apoptotic signal accounts for leading), and active cell occurs irreversible apoptosis.Research discovery tyrosine-kinase Enzyme inhibitor gefitinib (gefitinib)/Erlotinib (Erlotinib) treatment NSCLC patient, about 10% patient's performance Go out satisfied clinical effectiveness rapidly, further study show that these patient's overwhelming majority exist EGFR genetic mutation.At present The known gene mutation relevant with EGFR-TKI (endothelial growth factor receptor tyrosine kinase inhibitor) is confined to the most several: G719X (18 exon), E746-A450 lack (19 exon), L858R (21 exon), L861Q (21 exon), T790M (20 exon) and D770-N771 (20 exon).Wherein E746-A450 disappearance and the curative effect height with TKI that suddenlys change of L858R Relevant.Mitsudomi T, the Yatabe Y analysis result to 568 example Patients with Non-small-cell Lungs: at all nonsmall-cell lung cancers During in patient, the EGFR genetic mutation of about 90% concentrates on 19 or 21 exons, wherein outside the deletion mutation and 21 of 19 exons The patient of the point mutation in aobvious son takes the effective percentage of EGFR-TKI and all reaches more than 70%.Recent research prompting, outside EGFR Slotting human nature sudden change (D770-N771) of aobvious son 20 can make receptor that the sensitivity of EGFR--TKI is reduced by 100 times, the most also Find that the patient with this sudden change is inconspicuous to EGFR--TKI therapeutic response.The amplified production of extron 20 is carried out sub-clone Analyzing and find, T790M sudden change is the change that a base pair occurs from cytidine (C) to thymidine (T), at egg White level is exactly that the threonine in EGFR tyrosine kinase domain 790 site is replaced (T790M) by methionine, and this sudden change can make EGFR is in the state of being activated again, thus causes the acquired drug-resistance of TKI, and the reason of drug resistance is that sudden change causes EGFR structure to be sent out Changing, makes that TKI is in connection steric effect occurs.

Research prompting KRAS is had to be probably the reason of Gefitinib, Erlotinib initial drug-resistant. The Meta of Helenalinardou summarizes the TKI therapeutic effect of 1008 example NSCLC patients in analyzing, K-ras sudden change is occurring 165 patients in, the patient of 94% to TKI treat without significant reaction.In general, KRAS and EGFR sudden change NSCLC is phase Repel mutually. in different tumors subtypes, there is notable difference: EGFR sudden change be mainly seen in non-smoker, and KRAS is more It is common in the cancer that smoking is relevant.Because KRAS always betides in the NSCLC with Wild type EGFR, so being difficult to district Divide on earth be because KRAS insensitive to EGFR-TKI, be also because suddenling change without EGFR.

Vascular endothelial growth factor receptor (vascular endothelial growth factor receptor, VEGFR) family includes 3 kinds of hypotypes, it may be assumed that VEGFR-1 (F1t-1 can also be write), VEGFR-2 (KDR/Flk-1) simultaneously and VEGFR-3 (F1t 1), additionally, also 1 and 2 two cooperative expert systems of neuropilin (neuropilin).Wherein VEGFR-1 It is mainly distributed on vascular endothelial cell, hematopoietic stem cell, macrophage and mononuclear cell, can be with VEGF-A, VEGF-B and P1GF In conjunction with, the most relevant with the growth regulating of hematopoietic stem cell.VEGFR-2 is mainly distributed on vascular endothelial cell and lymphatic endothelia is thin In born of the same parents, can be combined with VEGF-A, VEGF-C, VEGF-D, VEGF-E.VEGF stimulating endothelial cell propagation, increase vascular permeability Property with the effect of new vascular generation mainly by combining and activating VEGFR-2 and realize. compared with VEGFR-2, VEGFR-1 with The affinity of VEGF is high 10 times, but the activity of regulation endotheliocyte is much lower, it may be possible to VEGFR-2 activity is had negative regulation Effect.VEGFR-3 mainly expresses at lymphatic endothelial cells, can be combined with VEGF-C and VEGF-D, regulates and controls lymphatic endothelium Growth.

Research shows: when diameter of tumor is more than 2mm, it is desirable to have new vessels provides nutrient substance and excretion metabolism to give up Thing.VEGF/VEGFR signal path plays key effect in tumor vascular generation, can be by blocking or interference VEGF/ The new life of VEGFR signal path suppression blood vessel, to reach to control the curative effect of the growth of tumor.With traditional cytotoxic drug phase Ratio, the antitumor drug with VEGF/VEGFR-2 as target has the biggest advantage. and under normal physiological conditions, angiogenesis only exists The physiological activity such as wound healing and menstrual cycle works, so using anti-angiogenic medicaments treatment tumor, to human body poison Property effect little, vascular endothelial cell directly contacts with blood, make medicine be more prone to arrive action site. by the most right The understanding of VEGF/VEGFR signal path mechanism of action, can obtain following several possible inhibitor research direction: a. and utilize Monoclonal antibody suppression VEGF or VEGFR so that it is can not be specific binding, disabling signal conducts.Gene can certainly be utilized Technology their expression of suppression, weakens its activity.B. design specific micromolecular inhibitor, be attached to VEGFR born of the same parents' outer VEGF knot Conjunction region, competitive antagonism VEGF, in like manner, it is also possible to be bonded to the particular combination territory of VEGFR on VEGF, competitive antagonism VEGFR.C. the intracellular kinase territory of VEGFR, the binding site of mainly ATP, competitively antagonism ATP so that it is Wu Fati are suppressed For phosphate.D. the critical proteins of the VEGFR downstream signal of intracellular is suppressed. in view of the compliance of patient, can be administered orally is little Molecule inhibitor is likely to be of good prospect.

Platelet derived growth factor (platelet.derived growth factor, PDGF) is induction and promotes blood Pipe formation effect the most by force, one of the most single-minded angiogenesis factor.PDGF mainly by combining with pdgf receptor (PDGFR), enters And activated protein kinase signal transduction pathway and play a role.PDGFR is made up of two kinds of subunits of α and β, has 3 kinds of dimers (PDGFR-α α, α β, β β), wherein β β dimer receptor (PDGFR-β) is mostly important, and its molecular weight is about 180～190ku, belongs to In tyrosine kinase receptor (receptor tyrosine kinase, RTK) family.PDGFR is in tumor formation and development process In also play an important role.The overexpression of PDGFR-β or overactivity all can stimulate intratumoral vasculature to generate, and promote tumor Growth.PDGFR-β is one of molecular marker of tumor vascular endothelial cell, high expressed in endothelial cells in tumor neogenetic blood vessels, And it is closely related with the growth of some tumor, transfer and prognosis.So PDGFR-β is an ideal neoplasm targeted therapy Target.

The Raf/MEK/ERK path of Raf kinases and mediation thereof has remarkable effect in tumour progression and transfer process, and Epidermal growth factor (EGF), VEGF (VEGF) and PDGF is included with many somatomedin (PDGF) etc. closely related.People thought multiple way to regulate this path, including suppression Ras albumen farnesyl Change, suppress Rat " expression of-I kinases (also referred to as C-RAF kinases), suppression Raf kinases and the kinase whose activity of MEK.Above-mentioned method Not only inhibit the signal transduction of ERK but also successfully inhibit the growth of xenograft tumours.Additionally, existing evidence shows, greatly Partial tumors is not arranged by single signal pathway, carries out suppressing to obtain bigger curative effect for Mutiple Targets.

Numerous disease is that the abnormal cell effect caused with protein kinase mediated event is associated.These disease bags Include, but be not limited to, tumor, inflammation disease, immunological diseases, osteopathia, metabolic disease, sacred disease, cardiovascular and cerebrovascular disease, hormone Relevant disease etc..Consequently found that be very important as medicine with finding kinases inhibitor.Send out although many Bright this area is made the biggest contribution, but for improving medication effect, research is still being continued in this area.

Summary of the invention

It is an object of the invention to provide 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropylidene hydrazine carbonyl) Pyridine-4-epoxide) phenyl) urea and 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropyl hydrazine carbonyl) pyridine-4- Epoxide) phenyl) urea, its officinal salt.

Another object of the present invention is to provide 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropylidene hydrazine carbonyl Base) pyridine-4-epoxide) phenyl) urea and 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropyl hydrazine carbonyl) pyridine- 4-epoxide) phenyl) preparation method of urea.

(((2-is different for 2-for 4-for-3-containing 1-(4-chloro-3-trifluoromethyl) to it is still another object of the present invention to provide one Propylidene hydrazine carbonyl) pyridine-4-epoxide) phenyl) urea and 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropyl hydrazine carbonyl Base) pyridine-4-epoxide) phenyl) pharmaceutical composition of this compound of urea.

A further object of the present invention be to provide this compounds anticancer, and with in the medicine of protein kinase related disorder Purposes.

In order to complete the purpose of the present invention, can adopt the following technical scheme that

The present invention relates to 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(the 2-isopropylidene hydrazine with having structure Carbonyl) pyridine-4-epoxide) phenyl) urea and 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropyl hydrazine carbonyl) pyrrole Pyridine-4-epoxide) phenyl) urea:

Or its officinal salt.

The invention also discloses the method preparing the compounds of this invention, comprise the steps:

1) route 1

A (), with hydrazides 1 as raw material, is readily available propylidene hydrazides 2 through common method and condensation of acetone.

(b) para hydroxybenzene amine under alkaline environment by the chlorine substituted ether in propylidene hydrazides 2 is obtained compound 3.

C () compound 3 can generate urea derivative I by the condensation of CDI Yu 4-chloro-3-5-trifluoromethylaniline；Also can be chloro-with 2- 4-NCO trifluoromethylbenzene obtains urea derivative I by nucleophilic addition.

D () urea derivative I is reducible obtains urea derivative II

2) route 2

A (), with ester 4 or acyl chlorides 5 as raw material, obtains hydrazide derivatives 7 with isopropyl hydrazine reaction.

(b) para hydroxybenzene amine under alkaline environment by the chlorine substituted ether in hydrazide derivatives 7 is obtained compound 8.

C () compound 8 can generate urea derivative II by the condensation of CDI Yu 4-chloro-3-5-trifluoromethylaniline；Also can be with 2- Chloro-4-NCO trifluoromethylbenzene obtains urea derivative II by nucleophilic addition.

3) route 3

A () para hydroxybenzene amine can generate urea derivative 9 by the condensation of CDI Yu 4-chloro-3-5-trifluoromethylaniline；Also can be with 2- Chloro-4-NCO trifluoromethylbenzene obtains urea derivative 9 by nucleophilic addition.

(b) urea derivative 9 under alkaline environment by hydrazide derivatives 7 chlorine nucleophilic displacement of fluorine etherificate obtain compound II；Also can be by the nucleophilic displacement of fluorine etherificate of chlorine in propylidene hydrazides 2 be obtained compound I under alkaline environment.

It addition, initiation material and intermediate in above-mentioned reaction are readily obtained, or to those skilled in the art Synthesis can be easy to by the conventional method in organic synthesis.

1-described in Formulas I and II (4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropyl subunit hydrazine carbonyl) pyridine-4- Epoxide) phenyl) urea and 4-(4-(3-(4-chloro-3-trifluoromethyl) urea groups) phenoxy group)-2-(2-Iproniazid carbonyl) pyridine, Described pharmaceutically acceptable salt includes different acid-addition salts, such as following mineral acid or the acid-addition salts of organic acid: hydrochloric acid, hydrogen bromine Acid, phosphoric acid, sulphuric acid, methanesulfonic acid, p-methyl benzenesulfonic acid, trifluoroacetic acid, Fructus Lycii acid, maleic acid, tartaric acid, fumaric acid, citric acid, breast Acid.Pharmaceutically acceptable salt described in I also includes Different Alkali slaine (lithium, sodium, potassium salt), alkali salt (calcium, magnesium salt) and Ammonium salt, and it is provided that the salt of the organic base of physiologically acceptable cation, such as methylamine, dimethylamine, trimethylamine, piperidines, morpholine And the salt of three (2-ethoxy) amine.All these salt within the scope of the present invention all can use conventional method to prepare.

The invention still further relates to the pharmaceutical composition using the compounds of this invention as active ingredient.This pharmaceutical composition can basis Prepared by method well known in the art.Can by by the compounds of this invention with-kind or multiple pharmaceutically acceptable solid or liquid Excipient and/or adjuvant combine, and make and are suitable to any dosage form that human or animal uses.The compounds of this invention is at its pharmaceutical composition In content be usually 0.1-95 weight %.

The compounds of this invention or the pharmaceutical composition containing it can be administered in a unit, and route of administration can be intestinal Or non-bowel, as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung and respiratory tract, skin, Vagina, rectum etc..

Form of administration can be liquid dosage form, solid dosage forms or semisolid dosage form.Liquid dosage form can be that solution (includes True solution and colloid solution), Emulsion (including o/w type, w/o type and emulsion), suspensoid, injection (include aqueous injection, injectable powder And transfusion), eye drop, nasal drop, lotion and liniment etc.；Solid dosage forms can be tablet (include ordinary tablet, enteric coatel tablets, buccal tablet, Dispersible tablet, chewable tablet, effervescent tablet, oral cavity disintegration tablet), capsule (including hard capsule, soft capsule, enteric coated capsule), granule, dissipate Agent, micropill, drop pill, suppository, membrane, paster, gas (powder) mist agent, spray etc.；Semisolid dosage form can be ointment, gel Agent, paste etc..

The compounds of this invention can make ordinary preparation, also to make be slow releasing preparation, controlled release preparation, targeting preparation and various Particulate delivery system.

In order to the compounds of this invention is made tablet, well known in the art various excipient can be widely used, including dilute Release agent, adhesive, wetting agent, disintegrating agent, lubricant, fluidizer.Diluent can be starch, dextrin, sucrose, glucose, breast Sugar, mannitol, sorbitol, xylitol, microcrystalline Cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate etc.；Wetting agent can be water, second Alcohol, isopropanol etc.；Binding agent can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline Cellulose, arabic gum Slurry, gelatine size, sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, card Ripple nurse, polyvinylpyrrolidone, Polyethylene Glycol etc.；Disintegrating agent can be dried starch, microcrystalline Cellulose, low substituted hydroxy-propyl fiber Element, crospolyvinylpyrrolidone, cross-linking sodium carboxymethyl cellulose, carboxymethyl starch sodium, sodium bicarbonate and citric acid, polyoxy second Alkene sorbitan fatty acid ester, dodecyl sodium sulfate etc.；Lubricant and fluidizer can be Pulvis Talci, silicon dioxide, tristearin Hydrochlorate, tartaric acid, liquid paraffin, Polyethylene Glycol etc..

Tablet can also be made coated tablet, such as sugar coated tablet, thin membrane coated tablet, ECT further, or double Synusia and multilayer tablet.

In order to administration unit is made capsule, effective ingredient the compounds of this invention can be mixed with diluent, fluidizer Close, mixture is placed directly within hard capsule or soft capsule.Also can be by first for effective ingredient the compounds of this invention and diluent, bonding Granule or micropill are made in agent, disintegrating agent, then are placed in hard capsule or soft capsule.For preparing each dilute of the compounds of this invention tablet Release agent, adhesive, wetting agent, disintegrating agent, fluidizer kind can also be used for preparing the capsule of the compounds of this invention.

For the compounds of this invention is made injection, water, ethanol, isopropanol, propylene glycol or their mixture can be used Make solvent and add solubilizing agent the most commonly used in the art, cosolvent, pH adjust agent, osmotic pressure regulator.Solubilizing agent or hydrotropy Agent can be poloxamer, lecithin, HP-β-CD etc.；PH adjustment agent can be phosphate, acetate, hydrochloric acid, hydrogen Sodium oxide etc.；Osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate etc..As prepared lyophilized powder Injection, also can add mannitol, glucose etc. as proppant.

Additionally, if desired, coloring agent, preservative, spice, correctives or other interpolation can also be added in pharmaceutical preparation Agent.

For reaching medication purpose, strengthening therapeutic effect, the medicine of the present invention or pharmaceutical composition can be given known to any Prescription method is administered.

The dosage of the compounds of this invention pharmaceutical composition is according to prevent or to treat the character of disease and serious journey Degree, patient or the individual instances of animal, route of administration and dosage form etc. can have change on a large scale.In general, of the present inventionization The Suitable dosage ranges of the every day of compound is 0.001-150mg/Kg body weight, preferably 0.01-100mg/Kg body weight.Above-mentioned dosage Can a dosage unit or be divided into several dosage unit administration, this depends on the clinical experience of doctor and includes using other The dosage regimen for the treatment of means.

The compound of the present invention or compositions can individually be taken, or merge use with other treatment medicine or symptomatic drugs. When the compound of the present invention exists synergism with other medicine, its dosage should be adjusted according to practical situation.

The compounds of this invention is Mutiple Targets kinases inhibitor or its precursor, and these protein kinases are according to phosphorylated substrate Difference be divided into multiple family, such as protein tyrosine kinase, Protein Serine/threonine kinase, lipoid etc..Typically, albumen Kinases transfers to a protein receptor relevant to signal transduction pathway by affecting a phosphoryl from a ribonucleoside triphosphote Carry out signal transduction in mediated cell.These phosphorylated events, as the biological function of molecular switch regulation target protein, are finally swashed Send out and reacted in various extracellulars and other stimulation.Kinases is present in multilayer signal transduction path, receptor tyrosine kinase It is positioned at upstream and the upstream of tumor cell Signal transduction pathway of tumor-blood-vessel growth Signal transduction pathway.Serine/threonine Protein kinase is positioned at the downstream of the Signal transduction pathway of tumor and tumor-blood-vessel growth cell.Research shows by blocking in upstream VEGFR and pdgf receptor, block Raf/MEK/ERK, it is possible to reduce the angiogenesis of tumor simultaneously and suppress tumor thin in downstream The duplication of born of the same parents, thus hinder the growth of tumor.The compounds of this invention has higher bioavailability, can be used for the multiple mankind and dislikes The treatment of property tumor, is hepatocarcinoma including described tumor disease, renal carcinoma, pulmonary carcinoma, cancer of pancreas, colorectal cancer, bladder cancer and mammary gland Cancer, ovarian cancer, squamous cell carcinoma, glioma, leukemia, incidence cancer.

Detailed description of the invention

Below with reference to embodiment, invention is described further, but does not limit the scope of the invention.

Determining instrument: NMR (Nuclear Magnetic Resonance) spectrum Vaariaan Mercury 300 type nuclear magnetic resonance analyser.Mass spectrum ZAD-2F With VG300 mass spectrograph.

Embodiment 1.4-chloro-N-propylidene picoline hydrazides

1.0 grams of 4-chlorine picoline hydrazides are joined in flask, adds 20 milliliters of acetone and be allowed to dissolve, add 1 ice vinegar Acid, under room temperature, stirring reaction 48h, TLC monitoring reaction terminates, and is concentrated by reactant liquor, adds 30 milliliters of acetic acid ethyl dissolutions, saturated Sodium bicarbonate solution washs, and anhydrous sodium sulfate is dried, and solvent evaporated obtains white solid 0.9 gram.¹H NMR(300MHz)(CDCl₃), 10.56 (S, 1H, CONH), 8.45 (d, 1H, ArH), 8.31 (S, 1H, ArH), 7.46 (d, 1H, ArH), 2.17 (S, 1H, CH₃), 2.04 (S, 1H, CH₃) .MS (FAB): (M⁺+ 1=212).

Embodiment 2.4-p-aminophenyl epoxide-N-propylidene picoline hydrazides

Under room temperature, 2.46 grams of para-aminophenol are dissolved in 20 milliliters of DMF, add 2.6 grams of potassium tert-butoxides, after stirring 2 hours Add 3.0 compounds and 0.09g K2C03 leads to heating 80 DEG C reaction 16h under nitrogen protective condition.After reaction terminates, it is cooled to room Temperature, is evaporated DMF, adds 10ml ethyl acetate and 10ml distilled water, extraction, and water layer 10ml ethyl acetate is extracted three times, merges Organic layer, washs three times with 50ml saturated aqueous common salt, and anhydrous Na 2SO4 is dried, and column chromatography for separation obtains violet solid,¹H NMR (300MHz) (DMSO-d6), 10.69 (S, 1H, CONH), 8.47 (d, 1H, ArH), 7.37 (S, 1H, ArH), 7.11 (d, 1H, ArH), 6.87 (d, 2H, ArH), 6.65 (d, 2H, ArH), 5.18 (S, 2H, NH₂), 2.00 (S, 1H, CH₃), 1.93 (S, 1H, CH₃)., MS (FAB) (M⁺+ 1=244)

Embodiment 3.1-is to chlorine m-trifluoromethylphenyl-3-(4-(2-propylidene hydrazine formylpyridine-4-oxygen) phenyl) urea

6.076 grams of CDI are dissolved in 20ML dichloromethane, add the DMAP of catalytic amount.Then under room temperature, dropping 6.642 grams is right 60 milliliters of dichloromethane solutions of chlorine 3-Aminotrifluorotoluene, after finishing, are stirred at room temperature 5 hours, and TLC monitoring raw material drips after disappearing Add the dichloromethane solution of 4-p-aminophenyl epoxide-N-propylidene picoline hydrazides, be warming up to 50 DEG C of reacting by heating after finishing 7 hours Stopped reaction, decompression removal solvent, column chromatography obtains product 4.0 grams.

¹H NMR (300MHz) (DMSO-d6), 10.72 (S, 1H, NH), 9.23 (S, 1H, NH), 9.02 (S, 1H, NH), 8.54 (d, 1H, ArH), 8.11 (S, 1H, ArH), 7.50-7.75 (m, 4H, ArH), 7.40 (S, 1H, ArH), 7.13-7.28 (m, 3H, ArH), 2.00 (S, 1H, CH₃), 1.94 (S, 1H, CH₃).MS(FAB)(M⁺+ 1=506)

Embodiment 4.1-is to chlorine m-trifluoromethylphenyl-3-(4-(2-Iproniazid formylpyridine-4-oxygen) phenyl) urea

0.04 gram of sodium cyanoborohydride is joined 0.3 gram of 1-to chlorine m-trifluoromethylphenyl-3-(4-(2-propylidene hydrazine first Acyl pyridine-4-oxygen) phenyl) urea dichloromethane solution in, and drip 0.037 milliliter of acetic acid, then under room temperature stirring to the most former Material disappears, and adds saturated sodium carbonate solution and terminates reaction, and dichloromethane extracts, and is dried, and rotation goes solvent to obtain product as white solid.

¹H NMR (300MHz) (DMSO-d6), 10.02 (d, 1H, NH), 9.61 (S, 1H, NH), 9.38 (S, 1H, NH), 8.49 (d, 1H, ArH), 8.13 (S, 1H, ArH), 7.55-7.70 (m, 4H, ArH), 7.34 (S, 1H, ArH), 7.10-720 (m, 3H, ArH), 4.84 (m, 1H, NH), 3.04-3.10 (m, 1H, CH), 0.98 (S, 1H, CH₃), 0.96 (S, 1H, CH₃).MS(FAB) (M⁺+ 1=508)

Pharmacologically active

External activity is evaluated:

Mtt assay measures tumor cell survival

It is 0.8～2 × 10 by being configured to concentration after the cell trypsinization of exponential phase⁴The Cell sap of cell/ml, Being inoculated in 96 orifice plates by 1000/hole, every hole adds 100 μ l.Next day add containing variable concentrations medicine and coordinative solvent comparison new Fresh culture medium, every hole adds 100 μ l (DMSO final concentration ＜ 0.5%), and every medicine sets 5～7 dosage groups, often organize at least set three parallel Hole, after continuing to cultivate 120hr in 37 DEG C, abandons supernatant, and every hole adds the freshly prepared serum-free training containing 0.5mg/ml MTT of 100 μ l Supporting base, continue to cultivate 4hr, abandon culture supernatant, every hole adds 200 μ l DMSO and dissolves MTT first hairpin precipitation, vibrates with microoscillator Mixing, measures optical density value (0D), with solvent by MK3 type microplate reader under the conditions of reference wavelength 450nm, detection wavelength 570nm The tumor cell of control treatment is matched group, calculates the suppression ratio of drug on tumor cell by formula below, and by middle efficacious prescriptions journey Calculate IC₅₀:

MTT the selection result (120h)

Activity in vivo is evaluated:

Impact on Renca tumor-bearing mice tumor growth

In laboratory animal entrance SPF level environment, breeding observing is after 24 hours, experiment of being allowed for access without exception.To exist in advance The Renca tumor liquid of Balb/c kind mouse peritoneal recovery, with aseptic normal saline by 1: 3 dilution proportion.Diluent is inoculated in Experiment mice left fore is subcutaneous, every injection dilution posterior tuberosity liquid 0.2mL.After the injection of all animals, empirically require random Packet, often group 8.

Start to be administered after inoculating 24 hours, once a day, gavage, it is administered 12 times altogether, dislocation execution after last is administered Animal, strips tumor tissues, and weighs.

Growth inhibited effect to mice-transplanted tumor Renca

* P ＜ 001；* * P ＜ 0.001, compares with negative control group.

#: animal is cause of fighting.

Claims (11)

1. Formulas I and 1 (4 chlorine 3 trifluoromethyl) 3 (4 (2 (the 2 isopropylidene hydrazine carbonyl) pyridine-4-epoxide) phenyl) urea shown in II With 1-(4-chloro-3-trifluoromethyl)-3-(4-(2-(2-isopropyl hydrazine carbonyl) pyridine 4 epoxide) phenyl) urea and pharmaceutically acceptable Salt

2. according to the officinal salt of claim 1 compound, it is characterised in that described officinal salt is selected from hydrochlorate, hydrogen bromine Hydrochlorate, phosphate, sulfate, mesylate, tosilate, acetate, trifluoroacetate, salicylate, amino acid salts, Fructus Lycii hydrochlorate, maleate, tartrate, fumarate, citrate, lactate.

3. the method for the described compound of preparation claim 1, comprises the steps:

With hydrazides 1 as raw material in step (a), it is readily available propylidene hydrazides 2 through common method and condensation of acetone；

In step (b), para hydroxybenzene amine under alkaline environment by the chlorine substituted ether in propylidene hydrazides 2 is obtained compound 3；

In step (c), compound 3 can generate urea derivative I by the condensation of CDI Yu 4-chloro-3-5-trifluoromethylaniline；Also can be with 2 Chlorine 4 NCO trifluoromethylbenzene obtains urea derivative I by nucleophilic addition；

In step (d), urea derivative I is reduced and provides urea derivative II.

4. the method for the described compound of preparation claim 1, comprises the steps:

In step (a), with ester 4 or acyl chlorides 5 as raw material, obtain hydrazide derivatives 7 with isopropyl hydrazine reaction；

In step (b), para hydroxybenzene amine under alkaline environment by the chlorine substituted ether in hydrazide derivatives 7 is obtained compound 8；

In step (c), compound 8 can generate urea derivative II by the condensation of CDI Yu 4-chloro-3-5-trifluoromethylaniline；Also can be with 2-chloro-4-NCO trifluoromethylbenzene obtains urea derivative II by nucleophilic addition.

5. the method for the described compound of preparation claim 1, comprises the steps:

In step (a), para hydroxybenzene amine can generate urea derivative 9 by the condensation of CDI Yu 4-chloro-3-5-trifluoromethylaniline；Also may be used 4-NCO trifluoromethylbenzene chloro-with 2-obtains urea derivative 9 by nucleophilic addition；

In step (b), urea derivative 9 under alkaline environment by the nucleophilic displacement of fluorine etherificateization of chlorine in hydrazide derivatives 7 Compound II；Also can be by the nucleophilic displacement of fluorine etherificate of chlorine in propylidene hydrazides 2 be obtained compound I under alkaline environment.

6. the compositions of a medicine, it is characterised in that compound containing any one of claim 1-2 and officinal salt thereof and Galenic pharmacy acceptable carrier.

7. compound and the officinal salt thereof of any one of claim 1-2 prevents the disease relevant with protein kinase with treatment in preparation The sick application in medicine.

8. compound and the officinal salt thereof of any one of claim 1-2 are relevant with tyrosine kinase with treatment in preparation prevention Application in the medicine of disease.

9. compound and the officinal salt thereof of any one of claim 1-2 prevents the disease relevant with Raf kinases with treatment in preparation The sick application in medicine.

Application the most according to claim 9, it is characterised in that the described disease relevant with Raf kinases is tumor, immunity mistake Tune, sacred disease.

11. application according to claim 10, it is characterised in that described tumor disease be hepatocarcinoma, renal carcinoma, pulmonary carcinoma, cancer of pancreas, Colorectal cancer, bladder cancer and breast carcinoma, ovarian cancer, squamous cell carcinoma, glioma, incidence cancer.