CN102584828B

CN102584828B - Tetramethyleneimine [3,4-d] pyrimidine derivatives, preparation method and application thereof

Info

Publication number: CN102584828B
Application number: CN201110008880.3A
Authority: CN
Inventors: 罗会兵; 郭建辉
Original assignee: Shanghai Allist Pharmaceuticals Inc
Current assignee: Shanghai Allist Medicine Polytron Technologies Inc
Priority date: 2011-01-14
Filing date: 2011-01-14
Publication date: 2016-01-27
Anticipated expiration: 2031-01-14
Also published as: CN102584828A

Abstract

The invention discloses a kind of tetramethyleneimine [3,4-d] pyrimidine derivatives, preparation method and application thereof.Tetramethyleneimine [3 of the present invention, 4-d] do be pyrimidine derivatives I type IGF-1 (type? I? insulin-like? growth? factor? receptor, IGF-1R) inhibitor, there is good suppression IGF-1R effect and cancer cell multiplication restraining effect, thus can be used as the therapeutical agent for the treatment of tumour and relative disease.

Description

Tetramethyleneimine [3,4-d] pyrimidine derivatives, preparation method and application thereof

Technical field

The present invention relates to a kind of suppression I type IGF-1 (typeIinsulin-likegrowthfactorreceptor, IGF-1R) active tetramethyleneimine [3,4-d] pyrimidine derivatives and preparation method thereof, and the application of this analog derivative on Therapeutic cancer and relative disease.

Background technology

Cancer is one of the most great disease threatening human life's health.In various disease, the mortality ratio of malignant tumour is in second, is only second to cardiovascular and cerebrovascular disease.Although along with the development of medical skill, considerable progress has been had with the anti-cancer therapies that operation and Radiotherapy chemotherapy are main method, but because the pathogenesis of cancer mechanism of action is complicated, treatment difficulty is very big, and therefore find small molecule anticancer drug that is efficient, low toxicity is one of the difficult point and focus of current field of cancer always.

I type IGF-1 (IGF-1R) is the receptor type tyrosine kinase (tyrosinekinese) very similar with the structure of insulin receptor (insulinreceptor), the assorted tetramer (YardenY etc. be made up of 2 extracellulars α subunit (subunit) and 2 cytolemma through β subunits, Biochemistry, 1988,27:3113-9).As I type or II type rhIGF-1 (insulin-likegrowthfactor-1,2 of its part; IGF-1, IGF-2), by being combined with its α subunit, making the β subunit activation with kinase domain (kinasedomain), causing IGF-1R to activate.By the IGF-1R that activated and then make I type and II type IRS (insulinreceptorsubstrate-1,-2) the multiple important substrate phosphorylation such as, via PI-3 kinase (phosphatidylinositol-3kinase), the signal path activation of mediation serine/threonine kinase Akt and blocking effect of mitogen activated protein kinases (mitogen-activatedproteinkinase, MAPK).Known Akt and MAPK signal path is responsible for important effect (Mitsiades etc., CurrentCancerDrugTargets, 2004,4:235) in transformation, propagation, existence, invasion and m etastasis.The generation of IGF-1R signal transduction pathway and tumour is closely related (PeruzziF etc., MolCellBiol, 1999,19:7203-15).As can be seen here, MAPK and the Akt signal path of IGF-1R Tyrosylprotein kinase and its Downstream regulatory plays an important role for the propagation of tumour cell, differentiation and transfer process.Therefore, the blocking-up of these signal paths is considered to the effective means of cancer treatment.

Massive epidemiology and clinical pathology experimental result show, in many tumour cells, and such as lung cancer, large bowel cancer, carcinoma of the pancreas, mammary cancer, prostate cancer, liver cancer, melanoma, multiple myeloma, leukemia etc., due to the rise of IGF-1, all there is overexpression in IGF-1R, height correlation (MichaelN.Pollak etc., NatureReviewCancer between its expression amount and tumorigenic probability, 2004,4:505).The overexpression of IGF-1R will promote the transfer (BasergaR etc., BiochimBiophysActa-RevCancer, 1997,1332:F105-106) of cancer cells further.

To sum up, IGF-1R is considered to the Effective target site of cancer treatment.Suppress the activity of IGF-1R, effectively can control growth and the transfer of tumour cell, and strengthen the susceptibility of tumours of chemotherapeutic, radiotherapy.The research and development of IGF-1R inhibitor becomes the focus (RongshiLi etc., J.Med.Chem., 2009,52:4981-5004) of antitumor area research.Be that in the methods for the treatment of of target, IGF-1R monoclonal antibody and small molecule kinase inhibitors enter clinical experimental stage with IGF-1R.

As IGF-1R micromolecular inhibitor, known compound structure has Pyrrolopyrimidine derivatives, Pyrazolopyrimidine derivative, imidazo pyrimidine derivatives, pyrazine benzimidazole derivative or benzimidizole derivatives etc.Wherein, the IGF-1R inhibitor OSI-906 of OSI company development has entered clinical III phase (MarkJMulvihill etc., FutureMed.Chem., 2009, 6:1153-1171), the IGF-1R inhibitor B MS-754807 of Bristol-Myers Squibb Co.'s development has entered clinical II phase (MarkD.Wittman etc., J.Med.Chem., 2009, 52:7360-7363), the IGF-1R inhibitor NVP-AEW-541 of Novartis Co., Ltd's development has entered clinical I phase (ToruMukohara, CancerLetters, 2009, 282:14-24), their structure is shown in following formula.

International patent application WO2008/021456 reports class dihydrofuran [3,4-d] pyrimidine derivatives and preparation method thereof, and discloses the compound of following pyrimidines structure,

They are disclosed for kinase activator agent and apoptotic inductor.

The international patent application WO2008005956A2 of Bristol-Myers Squibb Co. discloses the structure of following pyrroles's pyrrolotriazine derivatives

It is reported, these compounds are IGF-1R inhibitor, and they are disclosed and are used for the treatment of or preventing cancer and other diseases.

The international patent application WO2005097800 (CN200580017980.5) of OSI company discloses the compound of having structure

It is reported, these compounds are IGF-1R inhibitor, and they are disclosed and are used for the treatment of or prevent inflammatory diseases, autoimmune disorder, cancer and other diseases.

The international patent application WO02/092599 (CN02810003.4) of Novartis Co., Ltd discloses the compound of following Pyrrolopyrimidine derivatives

It is reported, these compounds are IGF-1R inhibitor, have antineoplastic activity.

WO98/31697 (Chinese patent publication No. 200680003323.X) discloses the indazole derivatives of having structure

It is reported, these indazole derivativess can as IGF-1R inhibitor.

In sum, IGF-1R is the cancer target with good DEVELOPMENT PROSPECT, and the specificity IGF-1R inhibitor effectively seeking high-efficiency low-toxicity has important clinical meaning and application prospect, and has become a large focus of current antineoplastic research.

Summary of the invention

The invention provides a kind of formula (I) compound, or its pharmacy acceptable salt,

Mark in formula represents following implication

R ¹for-CONHR ⁴, wherein R ⁴be selected from-(CH ₂) _0-6-aryl ,-(CH ₂) _0-6-heteroaryl or-(CH ₂) _0-6-nitrogenous saturated heterocyclyl, wherein said aryl, heteroaryl or nitrogenous saturated heterocyclyl is unsubstituted or quilt-halogen, C ₁~ C ₆alkyl, C ₁~ C ₆alkoxyl group, C ₃~ C ₈cycloalkyl ,-COOH ,-NH ₂or-OH replaces,

R ²be selected from-R ⁵,-halogen ,-OR ⁵,-COR ⁵,-COOR ⁵,-CONHR ⁵,-CH ₂nHR ⁵,-SOR ⁵,-SO ₂r ⁵or-CH ₂sO ₂r ⁵, wherein R ⁵for-H ,-C ₁~ C ₆alkyl ,-by the C of 1 ~ 3 halogen substiuted ₁~ C ₆alkyl ,-C ₃~ C ₈cycloalkyl ,-(CH ₂) _0-6-aryl ,-(CH ₂) _0-6-heteroaryl or-(CH ₂) _0-6-nitrogenous saturated heterocyclyl, wherein said aryl, heteroaryl or nitrogenous saturated heterocyclyl is unsubstituted or quilt-halogen, C ₁~ C ₆alkyl, C ₁~ C ₆alkoxyl group, C ₃~ C ₈cycloalkyl ,-COOH ,-NH ₂or-OH replaces,

R ³be selected from-H ,-halogen ,-C ₁~ C ₆alkyl ,-C ₃~ C ₈cycloalkyl ,-by the C of 1 ~ 3 halogen substiuted ₁~ C ₆alkyl ,-by the C of 1 ~ 3 halogen substiuted ₃~ C ₈cycloalkyl ,-NH ₂,-C=N ,-OH ,-COOH ,-CONH ₂,-(CH ₂) _0-6-aryl ,-(CH ₂) _0-6-heteroaryl or-(CH ₂) _0-6-nitrogenous saturated heterocyclyl, wherein said aryl, heteroaryl or nitrogenous saturated heterocyclyl are unsubstituted or quilt-halogen ,-C ₁~ C ₆alkyl or-C ₁~ C ₆alkoxyl group replaces,

N is 0,1,2,3 or 4.

In the present invention, term " aryl " refers to aromatic cyclic hydrocarbon group, and preferably carbonatoms is the aryl of 6 ~ 14, as benzene, naphthalene, is more preferably phenyl.

In the present invention, term " heteroaryl " refers to the 2 ring type heterocycles being selected from N, S, O heteroatomic 5-6 single heterocycle of unit or itself and phenyl ring condensing containing 1-4, and it is undersaturated.Here, as single heterocycle, preferably 5-6 unit bicyclic heteroaryl, particularly preferably be furyl, thienyl, pyrryl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazyl, thiadiazolyl group, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, be wherein more preferably imidazolyl, thiazolyl, triazolyl, pyridyl and pyrazinyl; As 2 ring type heterocycles, preferably benzofuryl, benzothienyl, diazosulfide base, benzothiazolyl, benzimidazolyl-, indyl, pseudoindoyl, indazolyl, quinolyl, isoquinolyl, quinazolyl.

In the present invention, term " nitrogenous saturated heterocyclic " refers to the 5-6 member heterocyclic ring containing nitrogen being selected from N, O or S atom containing 1-4, and it is saturated.Here, as nitrogen heterocyclic ring, preferably the nitrogenous saturated heterocyclic of 5-6 unit's monocycle, particularly preferably is pyrrolidyl, pyrazolidyl, imidazolidyl, piperidyl, morpholinyl, wherein more preferably pyrrolidyl, piperidyl or morpholinyl.

In the present invention, term " halogen " refers to fluorine, chlorine, bromine, iodine, preferred chlorine or fluorine.

In the present invention, term " C ₁-C ₆alkyl " refer to methyl, ethyl, propyl group, sec.-propyl, butyl, isobutyl-, sec-butyl, the tertiary butyl, amyl group, hexyl etc.; Preferable methyl, ethyl, propyl group, sec.-propyl or butyl.

In the present invention, term " C ₁-C ₆alkoxyl group " refer to methoxyl group, oxyethyl group, propoxy-, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert.-butoxy, pentyloxy, hexyloxy etc.; Preferred methoxyl group, oxyethyl group, propoxy-, isopropoxy or butoxy; More preferably methoxy or ethoxy.

In the present invention, term " C ₃-C ₈cycloalkyl " be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl or ring octyl group; Preferred cyclopropyl, cyclobutyl or cyclohexyl.

In the present invention's preferred embodiment, described tetramethyleneimine [3,4-d] pyrimidine derivatives or its pharmacy acceptable salt, represented by following formula general formula (II)

R in its formula of ², R ³, and R ⁴as described in as upper in general formula (I).

In the more preferred embodiment of the present invention one, the R described in general formula (II) compound ⁴for (CH ₂) _0-6-heteroaryl, described heteroaryl is unsubstituted or quilt-halogen, C ₁~ C ₆alkyl, C ₁~ C ₆alkoxyl group, C ₃~ C ₈cycloalkyl ,-COOH ,-NH ₂or-OH replaces; R ⁴be more preferably unsubstituted or quilt-halogen, C ₁~ C ₆alkyl, C ₁~ C ₆alkoxyl group ,-NH ₂or imidazolyl, thiazolyl, triazolyl, pyridyl or pyrazinyl that-OH replaces; R ⁴especially unsubstituted or quilt-halogen, C is preferably ₁~ C ₆alkyl, C ₁~ C ₆alkoxyl group ,-NH ₂or the pyridyl that-OH replaces; R ⁴most preferably be pyridyl that is unsubstituted or that be optionally substituted by halogen.

In the more preferred embodiment of the present invention one, the R described in general formula (II) compound ²for-COR ⁵,-COOR ⁵,-CONHR ⁵,-CH ₂nHR ⁵,-SOR ⁵,-SO ₂r ⁵or-CH ₂sO ₂r ⁵, wherein R ⁵for-H ,-C ₁~ C ₆alkyl, by the C of 1 ~ 3 halogen substiuted ₁~ C ₆alkyl ,-C ₃~ C ₈cycloalkyl, by-the C of 1 ~ 3 halogen substiuted ₃~ C ₈cycloalkyl ,-(CH ₂) _0-6-aryl ,-(CH ₂) _0-6-heteroaryl or-(CH ₂) _0-6-nitrogenous saturated heterocyclyl, wherein said aryl, heteroaryl or nitrogenous saturated heterocyclyl is unsubstituted or quilt-halogen, C ₁~ C ₆alkyl, C ₁~ C ₆alkoxyl group, C ₃~ C ₈cycloalkyl ,-COOH ,-NH ₂or-OH replaces.R ²be more preferably-COR ⁵, COOR ⁵or SO ₂r ⁵, wherein R ⁵for-H, C ₁~ C ₆alkyl, by the C of 1 ~ 3 halogen substiuted ₁~ C ₆alkyl or-(CH ₂) _0-6-aryl, described aryl is unsubstituted or quilt-halogen, C ₁~ C ₆alkyl, C ₁~ C ₆alkoxyl group or C ₃~ C ₈cycloalkyl substituted.R ⁵especially-H, C is preferably ₁~ C ₄alkyl, by the C of 1 ~ 3 halogen substiuted ₁~ C ₄alkyl, unsubstituted or quilt-halogen, C ₁~ C ₄alkyl or C ₁~ C ₄the benzene that alkoxyl group replaces.

In the more preferred embodiment of the present invention one, the R described in general formula (II) compound ³for unsubstituted or by the C of 1 ~ 3 halogen substiuted ₁~ C ₆alkyl, unsubstituted or by the C of 1 ~ 3 halogen substiuted ₃~ C ₈cycloalkyl.R ³be more preferably unsubstituted or by the C of 1 ~ 3 halogen substiuted ₁~ C ₄alkyl, unsubstituted or by the cyclopropyl of 1 ~ 3 halogen substiuted.

In the present invention, as the compound represented by general formula (I), concrete preferred compound is:

1-(4-(5-cyclopropyl-1H-pyrazoles-3-is amino)-6-(4-fluoro benzoyl)-6,7-dihydro-5H-pyrroles [3,4-d] pyrimidine-2-base)-N-(6-fluorine pyridin-3-yl) tetramethyleneimine-2-methane amide;

1-(6-ethanoyl-4-(5-cyclopropyl-1H-pyrazoles-3-is amino)-6,7-dihydro-5H-pyrroles [3,4-d] pyrimidine-2-bases)-N-(6-fluorine pyridin-3-yl) tetramethyleneimine-2-methane amide;

4-(5-cyclopropyl-1H-pyrazoles-3-is amino)-2-(2-(6-fluorine pyridin-3-yl-formamyl) pyrroles-1-base)-5H-pyrroles [3,4-d] pyrimidine-6 (7H)-t-butyl formate;

1-(4-(5-cyclopropyl-1H-pyrazoles-3-is amino)-6-(methyl sulphonyl)-6,7-dihydro-5H-pyrroles [3,4-d] pyrimidine-2-base)-N-(6-fluorine pyridin-3-yl) tetramethyleneimine-2-methane amide;

Or their pharmacy acceptable salts.

Present invention also offers the method that one prepares general formula (II) compound, it comprises the following steps:

Wherein, R ², R ³, R ⁴definition as described above.

With glycine ethyl ester hydrochloride (a) for starting raw material, add NaOH and ethyl propenoate, alkylated reaction obtains compound (b); Benzyl Chloride is added, K in compound (b) ₂cO ₃and NaI, after being dissolved in EtOH, condensation reaction obtains compound (c); Be dissolved in dry toluene by compound (c), then add potassium tert.-butoxide, annulation obtains compound (d); After compound (d) is molten with dehydrated alcohol, mix with urea and sodium ethylate, Cheng Huan obtains compound (e) again; Add phosphorus oxychloride at compound (e), chlorination obtains compound (f); Be dissolved in by compound (f) in 1,2-ethylene dichloride, drip triethylamine and Mono Chloro Acetic Acid chloroethene ester under ice bath, deprotection reaction obtains compound (g); After molten for Boc acid anhydrides DCM, be added drop-wise in compound (g), then add triethylamine, upper protecting group is obtained by reacting compound (h); After compound (h) is molten with DCM, add hydrochloric acid Isosorbide-5-Nitrae-dioxane, Deprotection again, is obtained by reacting compound (i); In compound (i), add THF, saturated sodium bicarbonate solution, then drip R ₂-Cl reagent, condensation reaction obtains compound (j); In compound (k), add Virahol and DIPEA, add compound (j) again under ice bath, substitution reaction obtains compound (l); In compound (l), add compound (m), DIPEA and N-Methyl pyrrolidone, substitution reaction obtains general formula (II) compound.

In above-mentioned each preparation process, the abbreviation of required reagent represents respectively:

Boc tertbutyloxycarbonyl

ClCH ₂cH ₂cl ethylene dichloride

DCM methylene dichloride

DIPEA diisopropyl ethyl amine

Et ₃n triethylamine

EtOH ethanol

HCl hydrochloric acid

K ₂cO ₃salt of wormwood

NaOH sodium hydroxide

NaHCO ₃sodium bicarbonate

NaI sodium iodide

NaOEt sodium ethylate

NH ₂c=ONH ₂urea

PhCH ₂cl Benzyl Chloride

POCl ₃phosphorus oxychloride

THF tetrahydrofuran (THF)

The present invention goes back inclusion compound pharmaceutically acceptable salt, and term " pharmacy acceptable salt " refers to acid salt or the base addition salt of the compounds of this invention of relative nontoxic.Described acid salt is the salt that the compounds of this invention and suitable mineral acid or organic acid are formed, these salt can be prepared in the last isolation andpurification process of compound, or the compound of purifying is reacted with its free alkali form and suitable organic acid or mineral acid, then the salt of formation is separated and makes.Representative acid salt comprises hydrobromate, hydrochloride, vitriol, sulphite, acetate, oxalate, valerate, oleate, palmitate, stearate, lauroleate, borate, benzoate, lactic acid salt, phosphoric acid salt, toluylate, Citrate trianion, maleate, fumarate, succinate, tartrate, benzoate, mesylate, tosilate, gluconate, Lactobionate and lauryl sulfonate etc.

Described base addition salt is the salt that the compounds of this invention and suitable mineral alkali or organic bases are formed, comprise the salt such as formed with basic metal, alkaline-earth metal, quaternary ammonium cation, as sodium salt, lithium salts, sylvite, calcium salt, magnesium salts, tetramethyl-quaternary ammonium salt, tetraethyl-quaternary ammonium salt etc.; Amine salt, comprises and ammonia (NH ₃), the salt that formed of primary amine, secondary amine or tertiary amine, as methylamine salt, dimethylamine salt, front three amine salt, triethylamine salt, ethylamine salt etc.

The compound of gained of the present invention or its pharmacy acceptable salt is utilized to deliver medicine to people, can oral, rectum, parenteral (intravenously, intramuscular or subcutaneous), topical (pulvis, ointment or drops).

Solid dosage for oral administration comprises capsule, tablet, pill, powder and granule.In these solid dosages, active compound mixes with at least one conventional inert excipients (or carrier), as Trisodium Citrate or Si Liaodengji dicalcium phosphate feed grade, or mix with following compositions: (a) filler or expanding material, such as, starch, lactose, sucrose, glucose, N.F,USP MANNITOL and silicic acid; (b) tackiness agent, such as, Walocel MT 20.000PV, alginate, gelatin, Polyvinylpyrolidone (PVP), sucrose and gum arabic; (c) wetting Agent for Printing Inks, such as, glycerine; (d) disintegrating agent, such as, agar, calcium carbonate, yam starch or tapioca (flour), alginic acid, some composition silicate and sodium carbonate; (e) retarding solvent, such as paraffin; F () absorbs accelerator, such as, and quaternary ammonium compound; (g) wetting agent, such as hexadecanol and glyceryl monostearate; (h) sorbent material, such as, kaolin; (i) lubricant, such as, talcum, calcium stearate, Magnesium Stearate, solid polyethylene glycol, sodium lauryl sulphate, or its mixture.In capsule, tablet and pill, formulation also can comprise buffer reagent.

Solid dosage such as tablet, sugar-pill, capsule, pill and granule can adopt dressing and the preparation of shell material, as casing and other material well known in the art.They can comprise opacifying agent, and in this composition, the release of active compound or compound can discharge in certain part in a delayed fashion in digestive tube.The example of adoptable embedding component is polymeric material and Wax.If desired, active compound also can form microencapsulation form with one or more in above-mentioned vehicle.

Liquid dosage form for oral administration comprises pharmaceutically acceptable emulsion, solution, suspension, syrup or tincture.Except active ingredient beyond the region of objective existence, liquid dosage form can comprise the conventional inert diluent adopted in this area, as water or other solvent, solubilizing agent and emulsifying agent, example is known, the mixture etc. of ethanol, Virahol, ethyl-carbonate, ethyl acetate, propylene glycol, 1,3 butylene glycol, dimethyl formamide and oil, particularly Oleum Gossypii semen, peanut oil, maize germ, sweet oil, Viscotrol C and sesame oil or these materials.

Except these inert diluents, composition also can comprise auxiliary agent, as wetting agent, emulsifying agent and suspension agent, sweeting agent, tender taste agent and spices.

Except active ingredient beyond the region of objective existence, suspension can comprise suspension agent, such as, and the mixture etc. of ethoxylation isooctadecane alcohol, polyoxyethylene sorbitol and Isosorbide Dinitrate, Microcrystalline Cellulose, aluminum methylate and agar or these materials.

Composition for parenteral injection can comprise physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, suspension or emulsion, and for being again dissolved into aseptic Injectable solution or the sterilized powder of dispersion liquid.Suitable moisture and nonaqueous carrier, thinner, solvent or vehicle comprise water, ethanol, polyvalent alcohol and suitable mixture thereof.

Formulation for the compounds of this invention of topical comprises ointment, powder, propellant and inhalation.Activeconstituents aseptically with physiologically acceptable carrier and any sanitas, buffer reagent, or the propelling agent that may need if desired is mixed together.

Present invention also offers a kind of pharmaceutical composition, it contains 0.1-500mg the compounds of this invention or its pharmacy acceptable salt, and pharmaceutically acceptable carrier, vehicle or thinner.

Present invention also offers a kind of method for the treatment of tumour, described tumour is alleviated by suppressing IGF-1R activity or is treated, and comprises step: the compounds of this invention or its pharmacy acceptable salt that use 0.1-50mg/kg body weight/day to the patient of needs treatment.

Compound of the present invention or its pharmacy acceptable salt can be individually dosed, or with other pharmaceutically acceptable therapeutic agent administrations, particularly with other anti-tumor disease drug regimens.Described therapeutical agent includes but not limited to: act on the drugs against tumor medicine of DNA chemical structure as cis-platinum, affect the antitumor drug of nucleic acid synthesis as methotrexate (MTX), 5 FU 5 fluorouracil (5FU) etc., affect the antitumor drug of transcribed nucleic acid as Zorubicin, pidorubicin, aclacinomycin, Plicamycin etc., act on the antitumor drug of tubulin synthesis as taxol, vinorelbine etc., arimedex is as aminoglutethimide, Lan Telong, letrozole, auspicious Ningde etc., cell-signaling pathways inhibitor is as epidermal growth factor receptor inhibitor imatinib (Imatinib), Gefitinib (Gefitinib), erlotinib (Erlotinib), lapatinibditosylate (Lapatinib) etc.Each composition to be combined can simultaneously or in a sequence give, and gives with unitary agent form or with the form of different preparation.Described combination not only comprises the combination of compound of the present invention and other promoting agent a kind of, and comprises the combination of compound of the present invention and two or more other promoting agents.

The compounds of this invention proves to have cancer cell multiplication restraining effect by cell experiment, can be used for the medicine preparing Therapeutic cancer.The drug effect using conventional procedures of the compounds of this invention anticancer propagation measures, a kind of preferred evaluation method is Sulforhodamine B (SulforhodamineB, SRB) protein staining method: SRB is a kind of protein-binding stain, can be combined by the basic aminoacids in biomacromolecule, its optical density(OD) at 510nm (OD) reading and protein content are good linear relationship, therefore can be used as the quantitative of cell count, calculate the inhibiting rate of medicine to cancer cell multiplication by the change measuring the absorbance value that drug effect produces after cancer cells.

Inhibiting rate (%)=(OD contrasts-OD inhibitor-OD blank)/(OD contrasts-OD blank) × 100%

OD contrasts: the OD value referring to the hole of the cell not having drug effect normal growth.

OD inhibitor: the OD value referring to the hole of the cell adding compound effects positive or to be screened.

OD blank: the OD value referring to the parallel control hole not having inoculating cell.

Half inhibitor concentration (IC ₅₀) value calculated by software GraphPadPrism5.

Prove that the compounds of this invention has IGF-1R inhibit activities by experiment, a kind of conventional experimental technique is protein immunoblotting test.Result shows that the compounds of this invention effectively can suppress the phosphorylation of IGF-1R, has IGF-1R inhibit activities.

Accompanying drawing explanation

Fig. 1 is embodiment 2 compound, and embodiment 3 compound and positive control drug (BMS-754807) suppress the protein immunoblotting test chart of IGF-1R phosphorylation.

Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.Unless otherwise indicated, otherwise number and per-cent are weight part and weight percent.

Embodiment

Embodiment 11-(4-(5-cyclopropyl-1H-pyrazoles-3-is amino)-6-(4-fluoro benzoyl)-6; 7-dihydro-5H-pyrroles [3,4-d] pyrimidine-2-base) synthesis of-N-(6-fluorine pyridin-3-yl) tetramethyleneimine-2-methane amide

Title compound is prepared by following step.

1) synthesis of compound (b):

Under ice bath, 300g glycine ethyl ester hydrochloride is dissolved in 450ml water, adds 4.7MNaOH (aq) 457ml, then add 227ml ethyl propenoate, stirred overnight at room temperature.After TLC detection reaction is complete, with DCM extractive reaction liquid, organic phase anhydrous Na ₂sO ₄drying, filters, revolves and steam to obtain crude yellow oily fluid cpds (b) 513.1g.

2) synthesis of compound (c):

By 513.1g compound (b), 392g Benzyl Chloride, 432gK ₂cO ₃, after 3.78gNaI 1LEtOH is molten, N ₂under protection, backflow is spent the night.After TLC detection reaction is complete, be cooled to room temperature, filter, be dissolved in after filtrate is spin-dried in ethyl acetate, washing, retain organic phase, wash with 2NHCl, water lotion saturated sodium bicarbonate is adjusted to PH=8, after be extracted with ethyl acetate, anhydrous Na SO ₄drying, filters, revolves and steam to obtain crude Compound (c) 1127.324g.

3) synthesis of compound (d):

Crude product 1127.324g compound (c) is dissolved in 2.5L dry toluene, 243.028g potassium tert.-butoxide is added gradually under ice bath, stir 2h, after TLC detection reaction is complete, PH is adjusted to be acid with 4MHCl under ice bath, layer saturated sodium bicarbonate of fetching water after layering is adjusted to PH=8, extraction into ethyl acetate, anhydrous Na ₂sO ₄drying, filters, revolves and steam to obtain 376.7g pink solid compound (d), yield 98%.

4) synthesis of compound (e):

After crude product 376.7g compound (d) is molten with 4L dehydrated alcohol, with urea 517.992g, after sodium ethylate 393.2g mixes, backflow is spent the night, and after TLC detection reaction is complete, spins off ethanol, add 1L water, stirring at room temperature 30min, with DCM extraction, retains aqueous phase, PH=3 is adjusted with 1L1NHCl (aq), filtering solids, washing, vacuum-drying.Obtain yellow solid compound (e) 34.1g, yield 30%.

5) synthesis of compound (f):

34.1g compound (e) is added phosphorus oxychloride 341ml, and backflow is spent the night.After TLC detection reaction is complete, cooling, slowly drips 1.5L saturated sodium bicarbonate solution under ice bath, is extracted with ethyl acetate, organic phase anhydrous Na ₂sO ₄drying, revolves and steams to obtain crude product red-brown oily matter compound (f) 11.388g.

6) synthesis of compound (g):

11.388g compound (f) is dissolved in 343ml1, in 2-ethylene dichloride, under ice bath, drips 5.22ml triethylamine, Mono Chloro Acetic Acid chloroethene ester 17.461g, backflow 8h.After TLC detection reaction is complete, mix with water 457ml, layering, organic phase anhydrous Na ₂sO ₄drying, is dissolved in 343ml methyl alcohol after being spin-dried for, backflow 1h, cooling, spins off methyl alcohol and obtain red brown solid compound (g) 10.4g.

7) synthesis of compound (h):

After molten for 17.67gBoc acid anhydrides DCM, be added drop-wise at 0 DEG C in compound (g), add triethylamine 23.417ml, after stirring at room temperature 3h, TLC detection reaction is complete, wash three times, organic phase anhydrous Na ₂sO ₄drying, is spin-dried for, column chromatography, with DCM: MeOH=20: 1 (v/v) for moving phase, obtains faint yellow solid compound (h) 4.12g, yield 23%.

8) synthesis of compound (i):

After 284mg compound (h) is molten with DCM, add 5.7ml hydrochloric acid Isosorbide-5-Nitrae-dioxane, after stirring at room temperature 1h, TLC detection reaction is complete, be spin-dried for obtain 250mg buff white solid powder compounds (i), yield 93%.

9) synthesis of compound (j):

116mg (0.610mol) compound (i) is added in 100ml single port bottle, 5.8mlTHF, 1.16ml saturated sodium bicarbonate solution, 0 DEG C drips fluorobenzoyl chloride 0.144ml (1.22mol), after TLC detection reaction is complete, revolves steaming, column chromatography, with sherwood oil: ethyl acetate=6: 1 (v/v) is moving phase, obtains 168mg compound as white solid (j), yield 33.2%.ESI(+)m/z：312

10) synthesis of compound (k):

Add in 100ml single port bottle under ice bath) 66.506mg1H-3-amino-5-cyclopropyl-pyrazoles (0.540mmol), add 4.1ml Virahol to make it to dissolve, add DIPEA0.17ml (1.026mmol), 168mg (0.540mmol) compound (j) is added under ice bath, stirred overnight at room temperature, after TLC detection reaction is complete, be cooled to 0 DEG C, filter, with a small amount of washed with isopropyl alcohol solid, obtain 131mg Beige powder (k) compound, yield 32.7%.

¹H-NMR(CDCl ₃，400MHz)：

7.65-7.68(m，2H)，7.33-7.36(m，2H)，6.43(s，1H)，4.50-4.64(m，4H)，1.93-1.99(m，1H)，0.92-0.98(m，2H)，0.70-0.75(m，2H)；

ESI(+)m/z：399

11) synthesis of compound 1:

(k) compound 131mg (0.421mol) is added in 100ml single port bottle, add N-(6-5-FU-3-base) tetramethyleneimine-2-acid amides 176mg (0.842mol), DIPEA0.18ml (1.095mol), N-Methyl pyrrolidone 5.24ml, DCM extracts, anhydrous Na ₂sO4 is dry, after being spin-dried for, climbing large plate and is separated, with ethylene dichloride: methyl alcohol: triethylamine=20: 1 (v/v) is moving phase, obtain red powder compound 129mg, yield 12.1%.

¹H-NMR(CDCl ₃，400MHz)：

δ8.33-8.38(m，1H)，8.12-8.15(m，1H)，7.63-7.68(m，2H)，7.30-7.36(m，2H)，7.13-7.16(m，1H)，6.25(s，1H)，4.55-4.67(m，4H)，3.78-3.85(m，2H)，3.53-3.61(m，1H)，2.24-2.28(m，1H)，1.90-2.04(m，3H)，1.74-1.78(m，1H)，0.81-0.92(m，2H)，0.75-0.79(m，1H)，0.65-0.73(m，1H)；ESI(+)m/z：572

The synthesis of embodiment 21-(6-ethanoyl-4-(5-cyclopropyl-1H-pyrazoles-3-is amino)-6,7-dihydro-5H-pyrroles [3,4-d] pyrimidine-2-bases)-N-(the fluoro-pyridin-3-yl of 6-) tetramethyleneimine-2-methane amide

With reference to the synthetic method of compound (j) in embodiment 1, replace fluorobenzoyl chloride with Acetyl Chloride 98Min. in the reactant of Intermediate Preparation, obtain compound 2 by the synthesis step of embodiment 1 compound.

1H-NMR(CDCl ₃，400MHz)：

δ8.34-8.37(m，1H)，8.13-8.17(m，1H)，7.17-7.20(m，1H)，6.23(s，1H)，4.58-4.69(m，4H)，3.72-3.78(m，2H)，3.56-3.64(m，1H)，2.28-2.33(m，1H)，2.15(s，3H)，1.90-2.04(m，3H)，1.74-1.78(m，1H)，0.81-0.92(m，2H)，0.75-0.79(m，1H)，0.65-0.73(m，1H)；

ESI(+)m/z：492

The synthesis of embodiment 34-(5-cyclopropyl-1H-pyrazoles-3-is amino)-2-(2-(6-fluorine pyridin-3-yl-formamyl) pyrroles-1-base)-5H-pyrroles [3,4-d] pyrimidine-6 (7H)-t-butyl formate

With reference to the synthetic method of compound (j) in embodiment 1, replace fluorobenzoyl chloride with tertiary butyl chloride manthanoate in the reactant of Intermediate Preparation, obtain compound 3 by the synthesis step of embodiment 1 compound.

¹H-NMR(CDCl ₃，400MHz)：

δ8.30-8.34(m，1H)，8.15-8.17(m，1H)，7.15-7.20(m，1H)，6.24(s，1H)，4.52-4.70(m，4H)，3.68-3.78(m，2H)，3.52-3.60(m，1H)，2.24-2.33(m，1H)，1.93-2.04(m，3H)，1.75-1.80(m，1H)，1.34(s，9H)，0.81-0.92(m，2H)，0.75-0.79(m，1H)，0.65-0.73(m，1H)；

ESI(+)m/z：550

Embodiment 41-(4-(5-cyclopropyl-1H-pyrazoles-3-is amino)-6-(methyl sulphonyl)-6; 7-dihydro-5H-pyrroles [3,4-d] pyrimidine-2-base) synthesis of-N-(the fluoro-pyridin-3-yl of 6-) tetramethyleneimine-2-methane amide

With reference to the synthetic method of compound (j) in embodiment 1, replace fluorobenzoyl chloride with Methanesulfonyl chloride in the reactant of Intermediate Preparation, obtain compound 4 by the synthesis step of embodiment 1 compound.

¹H-NMR(CDCl ₃，400MHz)：

8.32-8.35(m，1H)，8.14-8.20(m，1H)，7.15-7.20(m，1H)，6.24(s，1H)，4.46-4.57(m，4H)，3.68-3.78(m，2H)，3.52-3.60(m，1H)，2.87(s，3H)，2.24-2.33(m，1H)，1.93-2.04(m，3H)，1.75-1.80(m，1H)，0.81-0.92(m，2H)，0.75-0.79(m，1H)，0.65-0.73(m，1H)；

ESI(+)m/z：528

The preparation of embodiment 5 capsule

Compound 220g

Starch 140g

Microcrystalline Cellulose 65g

According to a conventional method, after being mixed by above-mentioned substance, load common gelatine capsule, obtain 1000 capsules.

By similar approach, the obtained capsule containing other embodiment compounds respectively.

Testing example 1 the compounds of this invention is to human breast cancer cell (MDA-MB-468) inhibited proliferation

The human breast cancer cell being in logarithmic phase is inoculated in 96 well culture plates with the density in about 5500/hole, 180 μ l/ holes.Each concentration establishes three wells.And establish the Vehicle controls of respective concentration and acellular zeroing hole.Adherent growth 24hr adds embodiment compound 20 μ l/ hole again, cell at 10%Hyclone foetal calf serum, 37 DEG C, cultivate 72hr under 5%CO2 condition.Add cold trichoroacetic acid(TCA) (TCA) 50ul of 50%, place 1 hour for 4 DEG C, fixed cell.Incline liquid, with the light and slow washing of distilled water 5 times, and seasoning in air.Add the SRB4mg/ml solution 100 μ l/ hole of being prepared by 1% Glacial acetic acid, dye 15 minutes in room temperature.Abandon supernatant liquor, wash 5 times with 1% acetic acid, air drying.Every hole adds the Tris solution (pH10.5) of the 10mM of 150 μ l, dissolves the SRB combined.Measuring OD value under microplate reader 510nm wavelength, obtaining the IC of embodiment compound for MDA-MB-468 cell by calculating ₅₀value:

Compound	IC ₅₀(μM)
		Embodiment 2	1.27
Embodiment 3	8.26
		Embodiment 4	8.45
Embodiment 5	25.38

Test result shows: the compounds of this invention has good inhibited proliferation to human breast cancer cell (MDA-MB-468).

Testing example 2 the compounds of this invention is to Non-small cell lung carcinoma cell (A549) inhibited proliferation

With reference to testing example 1 experimental technique, obtain the IC of embodiment compound for A549 cell by calculating ₅₀value:

Compound	IC ₅₀(μM)
		Embodiment 3	3.78

Test result shows: the compounds of this invention has good inhibited proliferation to Non-small cell lung carcinoma cell (A549).Testing example 3 the compounds of this invention is to human colon cancer cell (HT-29) inhibited proliferation

With reference to testing example 1 experimental technique, obtain the IC of embodiment compound for HT-29 cell by calculating ₅₀value:

Compound	IC ₅₀(μM)
		Embodiment 2	0.50
Embodiment 3	29.34
		Embodiment 4	0.81
Embodiment 5	5.88

Test result shows: the compounds of this invention has good inhibited proliferation to human colon cancer cell (HT-29).Testing example 4 the compounds of this invention is to human breast cancer cell (MCF-7) inhibited proliferation

With reference to testing example 1 experimental technique, obtain the IC of embodiment compound for MCF-7 cell by calculating ₅₀value:

Compound	IC ₅₀(μM)
		Embodiment 2	2.17
Embodiment 3	14.75
		Embodiment 4	2.33
Embodiment 5	19.69

Test result shows: the compounds of this invention has good inhibited proliferation to human breast cancer cell (MCF-7).

Testing example 5 protein immunoblotting is tested

To the human colon cancer cell HT-29 of logarithmic phase be in about 4 × 10 ⁵the density in individual/hole is inoculated in 6 well culture plates, adherent growth 24hr.Get nine holes, wherein add embodiment 2 compound (0.01 μM/hole in seven apertures in the human head respectively, 0.1 μM/hole and 0.5 μM/hole), embodiment 3 compound (0.01 μM/hole, 0.1 μM/hole and 0.5 μM/hole), positive control drug (BMS-754807,0.5 μM/hole), the not dosing of another holes.Cell at 10%Hyclone foetal calf serum, 37 DEG C, 5%CO ₂6hr is cultivated under condition.All add EGF in one hole of above-mentioned seven apertures in the human head and not dosing, stimulate 15 minutes.Cell, with after cold PBS washing, adds sample-loading buffer lysing cell.Cell lysate heats 10 minutes in 100 DEG C, and the centrifugal 5min of 12000g, gets supernatant.Supernatant carries out SDS-PAGE electrophoretic analysis.25mA constant current, wets and turns 2h, gone to by albumen on PVDF (polyvinylidinedifluoride) film.Take out pvdf membrane, with corresponding primary antibodie (pIGF-1R or pErk) 4 DEG C of overnight incubation.TBST washing vibration washing 3 times, each 10min.Add corresponding two again to resist, incubated at room temperature 1 hour.After TBST washs 3 times, with the development of ECL luminescence reagent box, and compressing tablet.Test-results is shown in accompanying drawing 1.

Test result shows: the compounds of this invention effectively can suppress the phosphorylation of IGF-1R, has IGF-1R inhibit activities.

The all documents mentioned in the present invention are quoted as a reference all in this application, are just quoted separately as a reference as each section of document.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims limited range equally.

Claims

1. the compound represented as following general formula (II) or its pharmacy acceptable salt,

In formula,

R ²be selected from-COR ⁵,-COOR ⁵or-SO ₂r ⁵, wherein R ⁵for-H or-C ₁~ C ₆alkyl,

R ³for unsubstituted or by the cyclopropyl of 1 ~ 3 halogen substiuted,

R ⁴be selected from-(CH ₂) _0-6-heteroaryl, wherein said heteroaryl is the pyridyl be optionally substituted by halogen.

2. compound as claimed in claim 1 or its pharmacy acceptable salt, wherein said R ²for-COR ⁵,-COOR ⁵or-SO ₂r ⁵, wherein R ⁵for-C ₁~ C ₆alkyl.

3. compound as claimed in claim 1 or its pharmacy acceptable salt, it is selected from:

Or their pharmacy acceptable salts.

4. medical composition, it comprises the carrier that compound described in any one of claims 1 to 3 or its pharmacy acceptable salt and pharmacopedics allow.

5. the application of the compound described in any one of claims 1 to 3 in preparation tumor.

6. prepare a method for formula according to claim 1 (II) compound, it comprises step:

Wherein, R ², R ³, R ⁴definition as claimed in claim 1,

With glycine ethyl ester hydrochloride (a) for starting raw material, add NaOH and ethyl propenoate, alkylated reaction obtains compound (b); Benzyl Chloride is added, K in compound (b) ₂cO ₃and NaI, after being dissolved in EtOH, condensation reaction obtains compound (c); Be dissolved in dry toluene by compound (c), then add potassium tert.-butoxide, annulation obtains compound (d); After compound (d) is molten with dehydrated alcohol, mix with urea and sodium ethylate, Cheng Huan obtains compound (e) again; Add phosphorus oxychloride at compound (e), chlorination obtains compound (f); Be dissolved in by compound (f) in 1,2-ethylene dichloride, drip triethylamine and Mono Chloro Acetic Acid chloroethene ester under ice bath, deprotection reaction obtains compound (g); After molten for Boc acid anhydrides DCM, be added drop-wise in compound (g), then add triethylamine, upper protecting group is obtained by reacting compound (h); After compound (h) is molten with DCM, add hydrochloric acid Isosorbide-5-Nitrae-dioxane, Deprotection again, is obtained by reacting compound (i); In compound (i), add THF, saturated sodium bicarbonate solution, then drip R ₂-Cl reagent, condensation reaction obtains compound (j); In Compound Compound (k), add Virahol and DIPEA, add compound (j) again under ice bath, substitution reaction obtains compound (l); In compound (l), add compound (m), DIPEA and N-Methyl pyrrolidone, substitution reaction obtains general formula (II) compound.