CN106890181A

CN106890181A - A kind of Foradil Aerolizer formoterol fumarate pharmaceutical composition of suppression VEGF

Info

Publication number: CN106890181A
Application number: CN201510977276.XA
Authority: CN
Inventors: 李静; 何光杰; 杨新意; 王淑丽; 陈立营; 胡筱芸; 孙亮
Original assignee: Tianjin Jinyao Group Co Ltd
Current assignee: Tianjin Jinyao Group Co Ltd
Priority date: 2015-12-21
Filing date: 2015-12-21
Publication date: 2017-06-27

Abstract

Foradil Aerolizer formoterol fumarate pharmaceutical composition the present invention relates to treat the breathing problem especially suppression VEGF of asthma, particularly containing active compound component I or its physiologically acceptable salt, one or more can be used for the dry powder Inhaled pharmaceutical composition of the excipient substance of inhalation powder spray.

Description

A kind of Foradil Aerolizer formoterol fumarate pharmaceutical composition of suppression VEGF

Technical field：

Medicine the present invention relates to treat breathing problem especially asthma, particularly containing active compound component I or Its physiologically acceptable salt, one or more dry powder Sucked medicines that can be used for the excipient substance of inhalation powder spray are combined Thing.

Background technology：

Angiogenesis includes two concepts：Brephic blood vessel generation (vasculogenesis, VG) and postnatal blood Pipe generates (ang1ogenesis, AG).VG is referred in the case of no vascular system, by endothelial progenitor cell (Endothelial progenitor cell, EPCs) or angioblast (angloblasts) are divided into endothelial cell, and Form rete vasculosum.AG is referred in adult blood, by the mature endothelial cell for having existed break through tube wall matrix migrate, propagation and Reconstruct, makes blood vessel branch persistently extend with Germination methods.Angiogenesis in the present invention refers to postnatal angiogenesis (anglogenesis, AG).Angiogenesis (anglogenesis) not still normal physiological change in (as growth, wound healing) Some processes, scientists are it has also been found that many diseases such as it and tumour, senile macular degeneration, malignant hematologic disease in recent years The development of disease has close relationship.Suppress pathologic angiogenesis, can treat or slow down tumour, senile macular degeneration, Many diseases such as malignant hematologic disease.

The method of current clinical treatment tumour is directed to different tumours using different methods mostly, and is mostly Treated for tumour cell.And tumour growth is a process for complexity, it is affected by various factors, including The foundation of tumor vessel net.Verified tumour growth has to rely on angiogenesis for many researchs, by suppressing angiogenesis Some links or whole process, and then the growth of tumour is controlled, to oncotherapy and prevent tumour DISTANT METASTASES IN significant. The beginning of the seventies depends on the proposition of vascularization concept with tumour growth, also accordingly proposes the general of anti-angiogenetic therapy treatment Read, i.e., by preventing the generation of new vessels and/or the extension of new vessels net and/or destruction new vessels from preventing small reality The generation or foundation of body knurl, to prevent tumour growth, development and shift.The more plus hydrogenated cortisone quilts of heparin of foreign countries' report It is known as effectively suppressing angiogenesis.It is experimentally confirmed that use in conjunction both it can suppress blood vessel on chick chorioallantoic membrane Generation, well can make tumor regression and prevent shift, may also suppress the rabbit corneal angiogenesis that tumour causes.

And VEGF (VEGF) is the multi-functional growth factor of a class, with promote endothelial cell proliferation, The effect that induction of vascular is formed, it is considered that suppressing VEGF (VEGF) can treat or slow down pathologic vessels It is newborn.

VEGF (VEGF) disease relevant with the new vessels such as disease such as tumour, bronchial astehma has close Relation is cut, by suppressing VEGF (VEGF), these diseases can be treated or slow down, lot of documents has this side The report in face is such as《Summarize the architectural feature and biological function of VEGF》(Chinese clinic study magazine, 2007 volume 13 3 Phase, 388),《The progress of VEGF and its acceptor in gynecological disease》(Hebei medicine 2006 volume 28 12 Phase, 1192-1194),《VEGF and Research Progress of relation between tumor》(Guangxi Medical University's journal, 2006 23 Roll up 2 phases, 333-335),《The present Research of VEGF correlation antineoplastons》(Jilin medical science, 5 phases of volume 27 in 2006,454- 457)、《Targeting VEGF treats the progress of malignant tumour》(modern medical oncology, 3 phases of volume 14 in 2006,370-372),《Blood Endothelial tube growth factor and bronchial astehma》(Chinese journals of practical medicine, volume 23 the 3rd phase, 433 in 2007).

Asthma is a kind of chronic airway inflammation, it is characterized by Reversible airway obstruction and airway reactivity increase, air flue resistance Plug is increased by the secretion that tunica mucosa bronchiorum inflammation causes, two kinds of factors of myxedema and inflammatory stimulus smooth muscle spasm are caused； And airway reactivity increases the result for being also due to the bronchial epithelial cell damage that airway inflammation causes.It is recognized that only There is the inflammation of control airway mucus, can be only achieved the final purpose for reducing airway hyperreactivity, alleviating SOA.Treatment at present The compound medicine of the medicine of the PUD Ds such as asthma commonly glucocorticoid and adrenaline broxaterol.

The content of the invention：

We have surprisingly found that the present invention is that formula (I) compound and salt have the effect for suppressing VEGF, especially medicinal Preparation aspect, after being made Foradil Aerolizer formoterol fumarate, the treatment tool especially for tumour, asthma has certain effect.

Those skilled in the art is believed that " treatment " involved by this paper can extend to the prevention of disease and with true Determine the treatment of disease.

A kind of inhalation powder spray pharmaceutical composition, it is characterised in that containing chemical compounds I or its physiologically acceptable salt, One or more can be used for the excipient substance of inhalation powder spray,

R=contains 3-10 carbon atom heteroatomic cycloalkyl, and wherein hetero atom is N, O, S.

Above-mentioned pharmaceutical composition, it is characterised in that can be used for inhalation powder spray is the one kind in carrier and/or additives Or it is several.Aforementioned pharmaceutical compositions, it is characterised in that the particle diameter of the carrier is 20~60 μm.Aforementioned pharmaceutical compositions, it is special Levy is that described carrier is one or more in carbohydrate carrier, amino acid, lecithin or phosphatid ylcholine.Said medicine group Compound, the amino acid is glycine, valine, leucine.Aforementioned pharmaceutical compositions, it is characterised in that the carbohydrate is single One or more of sugar, disaccharides or its derived carbohydrate, derived carbohydrate refer on glycan molecule at least one hydroxyl by comprising at most 20 carbon The straight or branched hydrocarbon chain substitution of atom.The monose is mannitol, fructose, glucose.The disaccharides is maltose, marine alga Sugar, cellobiose, lactose, sucrose.Described derived carbohydrate is the sugar ester of eight acetate fiber two, sucrose octa-acetate, eight acetic acid lactose Ester, five acetic acid glucose esters, six acetic acid Nitranitols and eight acetic acid marine alga sugar esters.

Aforementioned pharmaceutical compositions, it is characterised in that carrier is lactose.Described lactose is alpha-lactose monohydrate, β-anhydrous Lactose, one or more in amorphous spray-dried lactose, crystallizing and drying lactose.Described lactose is crystallizing and drying lactose.

Aforementioned pharmaceutical compositions, it is characterised in that described carrier is one or two in lecithin, phosphatid ylcholine And lactose.

Aforementioned pharmaceutical compositions, it is characterised in that additives are one or more in surfactant, lubricant.It is above-mentioned Pharmaceutical composition, it is characterised in that surfactant is poloxamer.The lubricant is magnesium stearate, superfine silica gel powder, talcum One or more in powder.

Above-mentioned pharmaceutical composition, it is characterised in that the substituent R of type I compound is to contain the 3-8 miscellaneous original of carbon atom Subring alkyl.

Above-mentioned pharmaceutical composition, it is characterised in that the hetero atom in the substituent R of type I compound is N.

Above-mentioned pharmaceutical composition, it is characterised in that the hetero atom in the substituent R of type I compound is O or S.

Above-mentioned pharmaceutical composition, it is characterised in that type I compound is：

5- amino -2- (2,6- difluorophenyls)-N- [5- (aziridine -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (azetidine -3- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (aza-cyclopentane -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (aza-cyclopentane -3- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (piperidine -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (piperidine -3- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (piperidine -4- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (azepan -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (azepan -- 3- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (azepan -- 4- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (Azacyclooctane -3- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (Azacyclooctane -5- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (oxirane -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (tetrahydrofuran -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (oxinane -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (oxepane -4- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (oxocane -4- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (Thietane -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (tiacyclopentane -3- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (thia hexamethylene -4- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (thia cycloheptane -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (thia cyclooctane -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine.

5- amino -2- (2,6- difluorophenyls)-N- [5- (Azacyclooctane -5- hydroxyls)-isothiazole -4-]-thiazole formyl Amine.

As claimed in claim 1 pharmaceutical composition, it is characterised in that type I compound is：

5- amino -2- (2,6- difluorophenyls)-N- [5- (azepan -2- hydroxyls)-isothiazole -4-]-thiazole formyl Amine.

5- amino -2- (2,6- difluorophenyls)-N- [5- (azepan -4- hydroxyls)-isothiazole -4-]-thiazole formyl Amine,

5- amino -2- (2,6- difluorophenyls)-N- [5- (oxocane -4- hydroxyls)-isothiazole -4-]-thiazole formyl Amine.

Above-mentioned pharmaceutical composition, it is characterised in that the physiologically acceptable salt of type I compound is from following sour shape Into salt：Hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, methanesulfonic acid, phenylacetic acid, fumaric acid, maleic acid, benzene sulphur One kind in acid, malic acid, glutamic acid, isethionic acid.

Above-mentioned pharmaceutical composition is used as the application in the medicine for the treatment of disease.

Above-mentioned pharmaceutical composition is used as the application in the medicine of Vascular endothelial growth factor receptor inhibitor.

Above-mentioned pharmaceutical composition as the Vascular endothelial growth factor receptor inhibitor by respiratory tract administration medicine In application.

Above-mentioned pharmaceutical composition is preparing the application for the treatment of lung tumors class disease medicament.

Application of the above-mentioned pharmaceutical composition in the medicine for preparing treatment respiratory inflammation.

Application of the above-mentioned pharmaceutical composition in the medicine for preparing treatment asthma.

Above-mentioned chemical compounds I or its physiologically acceptable salt are used as the application in the medicine for the treatment of disease.

Above-mentioned chemical compounds I or its physiologically acceptable salt as Vascular endothelial growth factor receptor inhibitor medicine Application in thing.

Above-mentioned chemical compounds I or its physiologically acceptable salt are the medicine for preparing treatment pathological disease Application in thing.

Neovascularization disease, relates generally to tumour, ocular angiogenesis, malignant hematologic disease, bronchial astehma, white matter of brain Loss disorders.

Tumour is mainly various entity tumors such as tumor stomach, lung tumors, liver's tumour, various body of gland tumours, nose Tumour, ocular tumor, pharynx tumor, laryngeal neoplasm etc.；Malignant hematologic disease refers to the malignant tumour of hematological system, main to include in vain Blood disease, lymthoma, Huppert's disease and malignant histiocytosis etc..

Above-mentioned chemical compounds I or its physiologically acceptable salt are preparing the application for the treatment of neoplastic diseases medicine.On The application of the chemical compounds I stated or its physiologically acceptable salt in the medicine for preparing treatment respiratory inflammation.

Above-mentioned chemical compounds I or its physiologically acceptable salt answering in the medicine for preparing treatment malignant hematologic disease With.

The application of above-mentioned chemical compounds I or its physiologically acceptable salt in the medicine for preparing treatment asthma.

A kind of method for preparing the type I compound that above-mentioned hetero atom is N：

Step one：

Under the normal temperature for adding within 8 carbon by compound 1 under nitrogen protection for the ether of liquid in, be then cooled to 0 DEG C with Under, n-BuLi is added dropwise thereto, after reaction terminates, to bromine is added dropwise in reaction solution, it is warmed to room temperature after being added dropwise to complete, stirring half is small When after be added thereto to hydrochloric acid solution, be layered, under water normal temperature for liquid material of organic ethers solvent extraction after discard, be associated with Machine phase, is concentrated to dryness after filtering, obtains yellow oily compounds 2；

Step 2：

By in the adding apparatus of compound 2, it is cooled to less than 0 DEG C, the nitration mixture (concentrated sulfuric acid that will be prepared in advance：The body of fuming nitric aicd Product is than being 55：36) it is added dropwise in reaction bulb, TLC display raw materials are cooled to room temperature after disappearing, and reaction solution is poured into trash ice, there is yellow Solid is separated out, filtering, and filter cake is washed with water to neutrality, is dried, and obtains the crude product of compound 3, gained crude product column chromatography after purification in vain Color solid chemical compound 3；

Step 3：

Under the normal temperature for adding within 8 carbon by compound 4 under nitrogen protection for the ether of liquid in, be subsequently adding NaH room temperatures Stirring 1 hour, is subsequently adding compound 3, reacts 2 hours, after TLC display raw materials disappear, to adding ethyl acetate in reaction solution And saturated aqueous common salt, organic phase anhydrous sodium sulfate drying after layering, dry, residue column chromatography purifying is concentrated under reduced pressure into after filtering Obtain yellow solid compound 6；

Step 4：

By in compound 6 and the 5%Pd/C 1-3 alcohol of carbon of addition, logical hydrogen carries out nitro-reduction reaction, and reaction was finished Filter, filtrate decompression is concentrated to give compound 7；

Step 5：

During compound 7 and compound 8 added into the 1-3 alcohol of carbon under nitrogen protection, heating response, reaction is finished, and decompression is dense It is reduced to dry, residue column chromatography purifies to obtain compound 9；

Step 6：Prepare compound I

Stirred during compound 9 is added into the 1-3 normal temperature liquified Halon of carbon, be subsequently adding HCl/ dioxane, stirred Reaction half an hour, PH is adjusted with organic base>Filtered after 7, obtain chemical compounds I.

A kind of method for preparing the type I compound that above-mentioned hetero atom is O or S：

Step one：

Step 2：

Step 3：

Step 4：

Step 5：

During compound 7 and compound 8 added into the 1-3 alcohol of carbon under nitrogen protection, heating response, reaction is complete,

It is concentrated under reduced pressure into dry, residue column chromatography purifies to obtain chemical compounds I.

A kind of salt method for preparing above-mentioned type I compound is that chemical compounds I is dissolved in into the halo that the 1-3 normal temperature of carbon is liquid In alkane, corresponding acid is dissolved under the normal temperature within 8 carbon in the ether or alcohol for liquid, and it is acidity, room temperature that pH value is adjusted after mixing Filtered after stirring half an hour, you can obtain the salt of chemical compounds I.

A kind of method for preparing the type I compound hydrochlorate that above-mentioned hetero atom is N：

Step one：

Step 2：

Step 3：

Step 4：

By in compound 6 and the 5%Pd/C 1-3 alcohol of carbon of addition, logical hydrogen carries out nitro-reduction reaction, instead

Filtering should be finished, filtrate decompression is concentrated to give compound 7；

Step 5：

Step 6：The salt of prepare compound I

The salt of 9 → chemical compounds I of compound

Stirred during compound 9 is added into the 1-3 normal temperature liquified Halon of carbon, be subsequently adding within the carbon of respective acids/5 Ether, regulation pH value is acidity, and stirring reaction half an hour, filtering obtains the salt of chemical compounds I.

Aforementioned pharmaceutical compositions, it is characterised in that described pharmaceutic adjuvant includes carrier and additives, the grain of the carrier Footpath is 20~60 μm, and preferable particle size is 30~60 μm.During carrier is carbohydrate carrier, amino acid, lecithin or phosphatid ylcholine One or more.The amino acid is glycine, valine, leucine.The carbohydrate is monose, disaccharides or its derived carbohydrate One or more, derived carbohydrate refer on glycan molecule at least one hydroxyl by comprising the at most 20 straight or branched hydrocarbon chains of carbon atom Substitution.The monose is mannitol, fructose, glucose.The disaccharides is maltose, trehalose, cellobiose, lactose, sucrose. Described derived carbohydrate is the sugar ester of eight acetate fiber two, sucrose octa-acetate, eight acetic acid lactose esters, five acetic acid glucose esters, six vinegar Sour Nitranitol and eight acetic acid marine alga sugar esters.Preferred vector is one or two in lactose, or lecithin, phosphatid ylcholine And lactose；Described lactose is alpha-lactose monohydrate, β-Lactis Anhydrous, amorphous spray-dried lactose, crystallizing and drying lactose In one or more, described lactose is more preferably crystallizing and drying lactose.

Described pharmaceutical composition, it is characterised in that additives are one or more in surfactant, lubricant.Surface Activating agent is poloxamer.Lubricant is one or more in magnesium stearate, superfine silica gel powder, talcum powder.

Aforementioned pharmaceutical compositions, the method for micronization of the active component uses spray drying process, fluid bed supersonic gas Stream comminuting method, speed lapping method, ball-milling method, fluid energy mill method or solvent method.

It should be appreciated that because it is known the reason for, such as retention of the active component in suction apparatus, every kind of work of patient's suction The amount of property composition can be differently configured from the amount of metering.Therefore the applicating ratio between active component may differ from the ratio of metering.It is excellent The ratio of the administration of choosing is in the described metered proportions for indicating.

Particle diameter such as nothing in the present invention refers in particular to be D90 mass median diameters (mass mean diameter), the particle diameter It is particle diameter corresponding when detecting that the cumulative particle sizes percentile of a sample reaches 90% by laser particle analyzer.

Specific embodiment

Present invention micronizing can use known mechanical crushing method or spray drying process.Mechanical crushing method refers to using stream Active component and carrier are ground into required particle diameter by the method that can be ground respectively.Spray drying process refers to by ciclesonide or load Body is dissolved in organic solvent such as ethanol entirely, by spray dryer, solid material is made into required particle diameter.It is dry using spraying Surfactant such as poloxamer etc. can also be added during dry method.

Capsule type dry powder inhalant can (sino-america joint-venture Suzhou Capsule Co., Ltd produces, commodity using No. 3 plant capsules Name：Vcaps), each embodiment feeds intake according to 1000 capsules.The capsule loading bubble-cap type of powder spray is aluminum-plastic packaged, when using Take out, load powder spray inhalator (Shanghai balance pharmaceutical factory) and use.

The mixed method of compound medicine：The small medicine of the taken amount medicine big with the amount of equivalent, while be placed in blender mixing Close, add with the big drug dilution of the amount of mixture equivalent uniformly, so a times amount is increased to untill adding the big component of whole amount, is mixed Even, the time of mixing is 10 minutes every time.

The method (equal increments method) that medicine mixes with auxiliary material：The auxiliary material component of thing and the equivalent of getting it filled, while being placed in mixing Mix in device, add and dilute uniform with the auxiliary material component of mixture equivalent, so a times amount is increased to and adds whole supplementary material component Untill, mixing, the time of mixing is 10 minutes every time.

Mixing machinery is using FGD stationary hoppers mixer (nine enginerring works of Shanghai day).

Embodiment 1

Specific embodiment：

Column chromatography method in the present invention：

The minimum 70cm of length of chromatographic column, inside filling 254- silica gel, and it is minimum that separate organic matter will be needed to be dissolved in entirely The chloroform of amount：Methyl alcohol=1：In 1, the top of silica gel in chromatographic column is placed in after the solution is absorbed with minimum 254- silica gel, made Eluted with mobile phase, the solution obtained by column chromatography is connect with several 10ml test tubes under chromatographic column, coutroi velocity is 10ml/ 3min, the solution of each test tube is analyzed with HPLC, and retention time identical test tube solution is merged, and takes the chemical combination of principal point Thing is recrystallized, and obtains corresponding product.

The method for determining principal point：The organic matter for needing separate is analyzed with HPLC, peak area is maximum in addition to raw material point Point be defined as principal point, its retention time for principal point retention time.

The method of the corresponding chipal compounds post of chiral post separation in the present invention：

Using CHIRALPAK AS-H particle diameter 5u 4.6mm (id) X 250mm analytical columns,

Mobile phase：N-hexane：Isopropanol：Ethylenediamine=90:9:0.1,

Detection wavelength：Ultraviolet 300 nanometers,

Column temperature：25℃

Flow velocity：1.0ml/min

RS compounds in embodiment are isolated the chipal compounds of R or S.

¹³C, 400MHz, DMSO-d6,1 to the 13 δ numerical value of carbon:

The position of C	1	2	3	4
					161.3-163.6	143.6-144.5	118.7-119.4	169.5-171.0
The position of C	5	6	7	8
					110.2-112.0	160.1-160.8	110.3-110.8	128.9-129.7
The position of C	9	10	11	12
					110.3-110.8	160.1-160.8	123.8-124.9	149.6-150.9
The position of C	13
					146.8-148.2

R in embodiment 1-1 to 1-22 sees the above table.

Embodiment 1-1 chemical compounds Is

Step one：

Compound 1 (20g, 0.235mol) is added into stirring and dissolving in dry 100ml ether under nitrogen protection, is then dropped Temperature is added dropwise n-BuLi (0.24mol) thereto to less than 0 DEG C, and less than 0 DEG C is maintained at during dropwise addition, is protected after being added dropwise to complete Temperature reaction is until without compound 1.

To bromine (0.5mol) is added dropwise in reaction solution, less than 0 DEG C is maintained at during dropwise addition, is slowly increased to after being added dropwise to complete Room temperature, is added thereto to hydrochloric acid solution (2N, 500ml) and reaction is quenched after stirring half an hour.

Layering, water is discarded after mutually being extracted with ether (200ml*3), merges organic phase, hydrogensulfite solution washing It is concentrated to dryness after (100ml*2), anhydrous sodium sulfate drying, filtering, obtains yellow oily compounds 2.

Ether can also be replaced with ethyl methyl ether, dimethyl ether, tetrahydrofuran, Isosorbide-5-Nitrae-dioxane equal solvent in this step.

Step 2：

By in the adding apparatus of 1mol compounds 2, it is cooled to less than 0 DEG C.Nitration mixture (the concentrated sulfuric acid that will be prepared in advance：Fuming nitric aicd Volume ratio be 55：36) it is slowly added dropwise in reaction bulb, TLC display raw materials are cooled to room temperature after disappearing, and reaction solution is poured into trash ice In, there is yellow solid to separate out.Filtering, filter cake is washed with water to neutrality, dries, and obtains the crude product of compound 3.Gained crude product column chromatography (ethyl acetate/n-hexane=1/20-1/10) obtains compound as white solid 3 after purification.

Step 3：

During 1mol compounds 4 are added into ether under nitrogen protection, it is subsequently adding 1.1molNaH and is stirred at room temperature 1 hour.Then After adding compound 3 (0.8mmol), reaction to be disappeared to TLC display raw materials, to addition ethyl acetate and saturated common salt in reaction solution Water, organic phase anhydrous sodium sulfate drying after layering, be concentrated under reduced pressure into after filtering it is dry, residue column chromatography (ethyl acetate/just oneself Alkane=1/10-1/5) purify to obtain yellow solid compound 6.

Step 4：

By in 1mol compounds 6 and 60g5%Pd/C addition methyl alcohol, logical hydrogen carries out nitro-reduction reaction, and reaction is to without original Filtering is finished in shots, reaction, and filtrate decompression is concentrated to give compound 7.

Methyl alcohol can also be replaced with ethanol, propyl alcohol equal solvent in this step.

Step 5：

During 1.3mol compounds 7 and 1mol compounds 8 added into 500ml methyl alcohol under nitrogen protection, heating response, reaction Finish, be concentrated under reduced pressure into dry.Residue column chromatography (ethyl acetate/n-hexane=1/5-1/2) purifies to obtain compound 9.

Step 6：

Stirred during 1mol compounds 9 are added into chloroform, be subsequently adding 2N HCl/ dioxane, stirring reaction half an hour, three Ethamine adjusts pH>Filtered after 7, dried with after chloroform, obtain chemical compounds I.

Chloroform can also be replaced with dichloromethane, 1,2- dichloroethanes equal solvent in this step.

The preparation method -1 of embodiment 1-1 hydrochlorides

0.1mol chemical compounds Is are dissolved in chloroform, are subsequently adding 2N HCl/1, in 4- dioxane, and PH are adjusted to low In after 5-6, it is concentrated under reduced pressure, filtering obtains the hydrochloride of chemical compounds I.

1,4- dioxane can also be replaced with ethyl methyl ether, dimethyl ether, tetrahydrofuran, ether equal solvent in this step.

The preparation method -2 of embodiment 1-1 hydrochlorides

Dissolved during 1mol compounds 9 are added into dichloromethane, 2N HCl/1 are subsequently adding, in 4- dioxane, after mixing Regulation pH value is 5-6, and stirring reaction is filtered after 0.5 hour, and filter cake is dried after being washed with methyl alcohol, obtains the hydrochloride of compound I.

1,4- dioxane can also be replaced with ethyl methyl ether, dimethyl ether, tetrahydrofuran, ether equal solvent in this step.This Chloroform can also be replaced with dichloromethane, 1,2- dichloroethanes equal solvent in step.

Embodiment 1-2 to 1-12

The preparation method of compound 1-2 to 1-12 with embodiment 1-1 preparation method.

The preparation method of embodiment 1-13

Embodiment 1-13 chemical compounds Is

Step one：

Step 2：

Step 3：

Step 4：

Step 5：

During 1.3mol compounds 7 and 1mol compounds 8 added into 500ml methyl alcohol under nitrogen protection, heating response, reaction Finish, be concentrated under reduced pressure into dry.Residue column chromatography (ethyl acetate/n-hexane=1/7-1/3) purifies to obtain chemical compounds I.

Embodiment 1-14 to 1-22

The preparation method of compound 1-14 to 1-22 with embodiment 1-13 preparation method

The preparation method of the salt of embodiment 1-2 to 1-22

The preparation method of the salt of compound 1-2 to 1-22 with embodiment 1-1 hydrochloric acid salt production process.

Above-mentioned salt is verified by elementary analysis

R in embodiment 1-1 to 1-22 sees the above table.

Embodiment 2-1 to 2-22：Inhalant

Active component is crushed respectively, particle diameter see the table below, the medicine after crushing mixes according to the method described above with auxiliary material respectively Afterwards according to active component 400 μ g/ be fitted into plant capsule.

Embodiment 3：Inhalant

Embodiment 4-1 to 4-22：Inhalation powder spray capsule

Active component：Embodiment 1-1 to 1-22 2g (5 μm of D50 particle diameters)

Auxiliary material：

Lactose：198g

Active component is diluted the pharmaceutical composition for being mixed into 200g with lactose in small, broken bits, is fitted into No. 4 hard shell capsules, medicine Dosage is 2mg/.The corresponding active components of embodiment 4-1 to 4-22 correspond to embodiment 1-1 to 1-22.

Embodiment 5-1 to 5-22：Inhalation powder spray capsule

Active component：2g (5 μm of D50 particle diameters)

Lactose：198g

Active component is diluted the pharmaceutical composition for being mixed into 200g with lactose in small, broken bits, is fitted into No. 4 hard shell capsules, medicine Dosage is 2mg/.

Pharmacological Examples 1：Experiment in vitro

Using chick chorioallantoic membrane (CAM) blood vessel hyperplasia model views compound of formula I of growth factor-induced to blood vessel The influence of hyperplasia, understands effect of the compound of formula I to angiogenesis.

Experiment material：

Fertilization instar chicken embryo on the 3rd

Glass fiber filter paper

Vascular endothelial growth factor (VEGF) Chem1con Products

Compound made by embodiment 1-13 is first dissolved with DMSO, then required concentration (DMSO is diluted to PBS Concentration<0.1%)

Experimental technique

The foundation of 1.CAM models：

75% ethanol cleaning chicken embryo chorion is dried up in super-clean bench, small at 100mm culture dishes edge after horizontal positioned 5min The heart breaks chorion into pieces, and ovum content is placed in culture dish (in advance added with 10ml DMEM culture mediums in culture dish).By this culture dish In being put into the big culture dishes of 150mm (in big culture dish plus little water), ware lid is covered, be placed in 37 DEG C, 5%CO₂Cell culture incubator Middle culture.

2. the influence pair CAM blood vessel hyperplasias

Glass fiber filter paper is made the sequin of diameter 3mm, moist heat sterilization, by each 10 μ l drops of reagent in glass with puncher On glass fiber filter paper, medicine film is made, dried up standby.After chick embryo culture 3d, random packet, every group 10, by administration group (1g/L) Give growth factor (2 μ g/L), and growth factor (VEGF, 2 μ g/L) individually stimulation group (positive controls) and PBS (phosphoric acid Salt buffer, pH 7.4) stimulation group (negative control group).By medicine film be affixed on CAM and yolk cyst membrane (yolk sacmembrane, YSM) the less position of blood vessel at 2/3 outward.After dosing 48h, basis of microscopic observation, large, medium and small blood in 5mm around counting medicine film Pipe number.

Table 1 to CAM blood vessel hyperplasias influence table (N=10)

Note：Statistical method data withCompare between representing groups using t inspections

3. result

The CAM that 1 compound of formula I is induced VEGF

The influence of blood vessel hyperplasia the results are shown in Table the good .VEGF groups thin vessels hyperplasia of 1, PBS group angiogenic growths substantially, Formulas I The each group blood vessel hyperplasia of compound is substantially suppressed thin vessels and substantially reduces, and returns to substantially normal level.

CAM is classical angiogenesis evaluation model, has the advantages that easy method, easy observation, inexpensive are most normal at present In vivo model.VEGF is important angiogenic factors, can stimulating endothelial cell hyperplasia and migration, promote blood vessel life Into, and overexpression is found in kinds of tumors tissue.The study show that：The CAM blood vessels that compound of formula I suppresses VEGF inductions increase It is raw, show that compound of formula I has the angiogenesispromoting effect for suppressing VEGF.

Compound of formula I has inhibitory action to angiogenesis, further demonstrate with the potentiality for suppressing Tumor Angiongesis.

Proved especially by experimental data, the compound containing azacyclo- and oxygen-containing or thia ring chemical combination in compound Thing is compared, and the angiogenesispromoting effect for suppressing VEGF is relatively low.

Pharmacological Examples 2：Internal anti-lung cancer experiment

1. experimental animal and knurl strain

Experiment is from Chinese large ear rabbit, 2.5 ± 0.1Kg of body weight, packet, every group 2.

2. lung cancer model preparation method

According to document《The foundation of rabbit VX2 lung cancer models and MR are observed》(packet header medical college journal, 2013 volume 29 the 2nd Phase, 7-9 pages) in method prepare rabbit VX2 lung cancer models.

3. treat and be grouped：Medicine is given by following table lattice

Active component prepares suction preparation according to the prescription and preparation method of embodiment 4, tests packet situation table

Note：Dosage presses active ingredient in embodiment test group.

4. administering mode

Be administered in 5 liters or so of cloche daily, experimental group be by medicine be fitted into suction apparatus according to suction Enter approach while giving medicine, control group gives the Lactis Anhydrous that average grain diameter is 55 μm according to inhalation route, dosage is 5mg/ Kg,

5. animal pattern Survival the average survival time situation comparative result

Table 2 sucks inhibitory action of the preparation to lung cancer

Group	The average survival time number of days	Group	The average survival time number of days
				Control group	28.5	22	51.5
DDP groups	56	23	43
				1	34.5	24	41.5
2	36	25	39
				3	38.5	26	38
4	37.5	27	44
				5	41	28	37.5
6	43	29	38.5
				7	42.5	30	39.5
8	45	31	41.5
				9	43.5	32	44
10	41.5	33	40.5
				11	31	34	38
12	31.5	35	37.5
				13	43	36	36
14	45	37	52
				15	45.5	38	48.5
16	47	39	48.5
				17	44	40	49
18	41	41	48
				19	49.5	42	51.5
20	43	43	47
				21	50	44	46

5 conclusions

Shown by experiment in vivo result, compound of formula I and its salt physiologically can significantly improve experiment lung cancer animal Life span, but improve the time the obvious difference of presence, proved especially by experimental data, azepine is contained in compound The compound of ring and oxygen-containing or thia ring compound phase ratio, suppress tumour growth effect relatively low.

The asthma pharmacological evaluation of Pharmacological Examples 3

1st, animal model

Choose healthy male Wistar rat (to check there is bacterium, the rat of bacterium infection refuses to select), body weight It is 200 ± 10g, is put into 5 liters or so of cloche, 3% chlorination second phthalein choline and 0.1% phosphorus is sprayed into the pressure of 400mmHg Sour histamine volume mixed liquor 15 seconds.After spraying stops, (there is asthma, breathing extremely in the asthmatic latent period for observing rat Difficulty, until twitching the time fallen), draw and breathe heavily the latent phase and refuse to select less than 70 seconds or the rat more than 120 seconds.

2nd, experimental technique

Learn from else's experience and determine the qualified rat of asthmatic latent period, be grouped at random according to the group in following table, every group 10, often It is administered in 5 liters or so of cloche, and experimental group is by the medicine dress in embodiment 5-1 to 5-22 and 4-1 to 4-22 Enter in suction apparatus according to inhalation route while giving medicine, the μ g/kg of dosage 500, model group 1 gives according to inhalation route Average grain diameter is 55 μm of Lactis Anhydrous, and dosage is 5mg/Kg, and the 10th day of administration is sprayed after whole medicines are given after 1.5 hours Mist gives 0.3% 2 hydrochloric acid histamine, and the change of asthmatic latent period and tic incidence (draws animal when breathing heavily before and after observation drug Tumble person is occurred without in 6 minutes and breathes heavily the volt phase as calculating in 360 seconds to draw).

3rd, experimentation and result：There is asthma in animal, up to twitching, the time fallen see the table below.

There are multiple forms to be the result of this experiment in the present embodiment, for convenience of description problem, experimental result is carried out Segmentation.

Table 1-1 embodiments experimental result (n=10, mean ± SD)

Show that compound of formula I can improve the animal asthmatic latent period time by the experiment in vivo result of Pharmacological Examples 3, The asthmatic latent period of embodiment group is compared with the asthmatic latent period of model group, and both gaps are obvious, so formula I is logical The generation of suppression new vessels is crossed, the effect with respiratory tract infection diseases such as treatment asthma, chronic obstructive pneumonias.

Proved especially by experimental data, in compound of formula I and its salt physiologically the compound containing azacyclo- with Oxygen-containing or thia ring compound phase ratio, the raising animal asthmatic latent period time is of a relatively high, and curative effect is substantially good；And in nitrogen-containing hetero 5-1,5-4,5-7,5-9,5-10,5-11,5-12 pharmacological action are better than other in the compound of ring.

Be may certify that by the data in Pharmacological Examples 1,2,3, compound of formula I and its salt physiologically are to angiogenesis It is inhibited with VEGF, the disease relevant with this can be treated or prevent, growth and the propagation of tumour can be suppressed, improve Asthmatic latent period etc..

Claims

1. a kind of inhalation powder spray pharmaceutical composition, it is characterised in that contain chemical compounds I or its physiologically acceptable salt, Kind or it is several can be used for the excipient substance of inhalation powder spray,

2. pharmaceutical composition as claimed in claim 1, it is characterised in that can be used for inhalation powder spray for carrier and/or additives In one or more.

3. pharmaceutical composition as claimed in claim 1, it is characterised in that type I compound is：

5- amino -2- (2,6- difluorophenyl)-N- [5- (aziridine -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (azetidine -3- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (aza-cyclopentane -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (aza-cyclopentane -3- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (piperidine -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (piperidine -3- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (piperidine -4- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (azepan -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (azepan -- 3- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (azepan -- 4- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (Azacyclooctane -3- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyls)-N- [5- (Azacyclooctane -5- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (oxirane -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (tetrahydrofuran -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (oxinane -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (oxepane -4- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (oxocane -4- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (Thietane -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (tiacyclopentane -3- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (thia hexamethylene -4- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyl)-N- [5- (thia cycloheptane -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides,

5- amino -2- (2,6- difluorophenyls)-N- [5- (thia cyclooctane -2- hydroxyls)-isothiazole -4-]-thiazole carboxamides.

4. pharmaceutical composition as claimed in claim 1, it is characterised in that the physiologically acceptable salt of type I compound be from The salt that following acid is formed：Hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, methanesulfonic acid, phenylacetic acid, fumaric acid, Malaysia One kind in acid, benzene sulfonic acid, malic acid, glutamic acid, isethionic acid.

5. pharmaceutical composition as claimed in claim 1 is used as the application in the medicine for the treatment of disease.

6. chemical compounds I as claimed in claim 1 or its physiologically acceptable salt are used as vascular endothelial growth factor receptor Application in the medicine of inhibitor.

7. chemical compounds I as claimed in claim 1 or its physiologically acceptable salt are preparing treatment pathological Application in the medicine of disease.

8. chemical compounds I as claimed in claim 1 or its physiologically acceptable salt are preparing treatment neoplastic diseases medicine Application.

9. chemical compounds I as claimed in claim 1 or its physiologically acceptable salt are preparing the medicine for the treatment of respiratory inflammation Application in thing.

10. chemical compounds I as claimed in claim 1 or its physiologically acceptable salt are preparing the medicine for the treatment of malignant hematologic disease Application in thing.

11. chemical compounds Is as claimed in claim 1 or its physiologically acceptable salt are in the medicine for preparing treatment asthma Using.

12. a kind of to prepare hetero atom in claim 1 be the method for the type I compound of N：

Step one：

Under the normal temperature for adding within 8 carbon by compound 1 under nitrogen protection for the ether of liquid in, be then cooled to less than 0 DEG C, to N-BuLi is wherein added dropwise, after reaction terminates, to bromine is added dropwise in reaction solution, is warmed to room temperature after being added dropwise to complete, after stirring half an hour Hydrochloric acid solution is added thereto to, is layered, to be discarded after the material of organic ethers solvent extraction of liquid under water normal temperature, merged organic Phase, is concentrated to dryness after filtering, obtains yellow oily compounds 2；

Step 2：

By in the adding apparatus of compound 2, it is cooled to less than 0 DEG C, the nitration mixture (concentrated sulfuric acid that will be prepared in advance：The volume ratio of fuming nitric aicd It is 55：36) it is added dropwise in reaction bulb, TLC display raw materials are cooled to room temperature after disappearing, and reaction solution is poured into trash ice, there is yellow solid Separate out, filtering, filter cake is washed with water to neutrality, dry, obtain the crude product of compound 3, gained crude product column chromatography after purification it is white solid Body compound 3；

Step 3：

Under the normal temperature for adding within 8 carbon by compound 4 under nitrogen protection for the ether of liquid in, be subsequently adding NaH and be stirred at room temperature 1 Hour, compound 3 is subsequently adding, react 2 hours, after TLC display raw materials disappear, to addition ethyl acetate and saturation in reaction solution Saline solution, organic phase anhydrous sodium sulfate drying after layering is concentrated under reduced pressure into dry after filtering, residue column chromatography purifies to obtain yellow Solid chemical compound 6；

Step 4：

By in compound 6 and the 5%Pd/C 1-3 alcohol of carbon of addition, logical hydrogen carries out nitro-reduction reaction, and filtering, filter are finished in reaction Liquid is concentrated under reduced pressure to obtain compound 7；

Step 5：

During compound 7 and compound 8 added into the 1-3 alcohol of carbon under nitrogen protection, heating response, reaction is complete, is concentrated under reduced pressure into Dry, residue column chromatography purifies to obtain compound 9；

Step 6：Prepare compound I

Stirred during compound 9 is added into the 1-3 normal temperature liquified Halon of carbon, be subsequently adding HCl/ dioxane, stirring reaction Half an hour, PH is adjusted with organic base>Filtered after 7, obtain chemical compounds I.

13. a kind of to prepare hetero atom in claim 1 be the method for the type I compound of O or S：

Step one：

Step 2：

Step 3：

Step 4：

Step 5：

During compound 7 and compound 8 added into the 1-3 alcohol of carbon under nitrogen protection, heating response, reaction is complete, is concentrated under reduced pressure into Dry, residue column chromatography purifies to obtain chemical compounds I.

A kind of 14. salt methods for preparing claim 1 type I compound are that chemical compounds I is dissolved in into the 1-3 normal temperature of carbon for liquid In alkyl halide, corresponding acid is dissolved under the normal temperature within 8 carbon in the ether or alcohol for liquid, and it is acidity that pH value is adjusted after mixing, Filtered after half an hour is stirred at room temperature, you can obtain the salt of chemical compounds I.

15. a kind of to prepare hetero atom in claim 3 be the method for the physiologically acceptable salt of type I compound of N：

Step one：

Step 2：

Step 3：

Step 4：

Step 5：

Step 6：The salt of prepare compound I

The salt of 9 → chemical compounds I of compound

Stirred during compound 9 is added into the 1-3 normal temperature liquified Halon of carbon, be subsequently adding the ether within the carbon of respective acids/5, Regulation pH value is acidity, and stirring reaction half an hour, filtering obtains the salt of chemical compounds I.