CN105121442A - PI3K and/or mTOR inhibitor prodrug - Google Patents

PI3K and/or mTOR inhibitor prodrug Download PDF

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CN105121442A
CN105121442A CN201480010513.9A CN201480010513A CN105121442A CN 105121442 A CN105121442 A CN 105121442A CN 201480010513 A CN201480010513 A CN 201480010513A CN 105121442 A CN105121442 A CN 105121442A
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CN105121442B (en
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吴永谦
薛建军
李琳
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Shandong Xuanzhu Pharma Co Ltd
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    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
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    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings

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Abstract

Provided are a compound as represented by general formula (I), pharmaceutically acceptable salt and stereoisomer thereof, and pharmaceutical composition containing the compound of general formula (I). Also provided are uses of the compound, pharmaceutically acceptable salt and stereoisomer thereof in the preparation of drugs for treating and/or preventing proliferative diseases.

Description

PI3K and/or mTOR inhibitor prodrug
PI3K and/or mTOR inhibitors prodrug technologies field
Pharmaceutical composition the present invention relates to the prodrug of PI3K and/or mTOR inhibitors, its pharmaceutically acceptable salt and their stereoisomer and comprising these compounds.Application the invention further relates to the preparation method of aforementioned prodrugs compound and its in the medicine for preparing treatment and/or prevention proliferative diseases.Background technology
Tumour is body under the effect of the various tumorigenesis factors, causes cellular genetic material to change, causes gene expression not normal, neoformation formed by abnormal cell proliferation.Tumour cell loses normal growth regulatory function, with autonomous or relative autonomous growth ability, when remaining to continued growth after the stopping of the tumorigenesis factor, largely the nutriments of consumption human bodies.If it find that with treatment not in time, cancer cell also can be transferred to whole body growth and breeding everywhere, and discharge a variety of toxin, human body is caused to be become thin, anaemia, organ function are damaged or even dead.
The method of tumour is treated mainly comprising three aspects：Drug therapy, operative treatment and radiotherapy.Due to operative treatment, radiotherapy be difficult to it is thoroughly just blunt remove tumour, and the effect of centering patients with advanced cancer is not obvious, therefore status of the drug therapy in oncotherapy is more and more obvious.Traditional anti-tumor medicine cannot be distinguished by tumour cell and normal tissue cell, often result in serious side effect, targeted drug is used as specific target spot using cancer cell, tumour can accurately be acted on, greatly improve treatment level, and adverse reaction rate is reduced, for example making the median survival time of advanced CRC increases by 66.7%, and the treated effect of advanced breast cancer improves 71.3%.
Because each drugmaker accelerates to the development for targetting class antineoplastic, along with market is in strong demand to the antineoplastic of this classification, molecular targeted agents have become fastest-rising unit in global antineoplastic market.Phosphatidylinositol3 3 kinase（PI3K) path is the place that variation is occurred most frequently in human cancer cell, cell can be caused to breed, and is activated, amplified signal.PI3K and mammal rapamycin target protein（MTOR) be PI3K signal paths important kinases.
PI3K is fat kinase families member, can produce Phosphatidyl inositol triphosphate fat by 3 phosphorylations of phosphatidyl alcohol（PIP3 cell metabolism and growth are adjusted).The second messenger PIP3 of the lipid can make P13K and the effector in downstream（Particularly Akt) oneself is to combining, so that cause film to be raised and phosphorylation, cell propagation, activation.Therefore suppress PI3K kinases, PI3K paths can be influenceed, so as to suppress cancer cell multiplication, activate. MTOR is a kind of protein serine/threonine being present in endochylema, belongs to Phosphoinositide-3 kinase related protein kinase enzyme family, is existed in vivo in the form of two kinds of compounds, i.e. mTORCl (the action target spots of rapamycin）(do not suppressed with mTORC2 by rapamycin）.MTOR is a kind of cell signalling albumen, and it adjusts reaction of the tumour cell to nutrient and growth factor, and passes through the blood supply of the effect control tumour to VEGF.MTOR inhibitor can make cancer cell hungry, and make tumor mass reduction by the effect for suppressing mTOR.
In the patent application WO2006122806 of Novartis Co., Ltd and the patent application WO201003816 of Pfizer, it was recently reported that have the series compound of inhibitory action to PI3K and mTOR, these compounds have good oncotherapy activity.Jo urnal of Medicinal Chemistry (2011), 54 (5), 1473-1480, " Discovery of 9- (6-Aminopyridin- 3-yl)-l-(3- (trifluoromethyl) phenyl) benzo [h] [l, 6] naphthyridin-2 (l H)-one (Torin2) as a potent, selective, and orally available mammalian target of rapamycin (mTOR) inhibitor for treatment of cancer. disclose entitled Torin2 compound, and report result of study to its internal pharmacokinetics.
But such compound mostly haves the shortcomings that poorly water-soluble, cause difficult to patient's conveying medicine.Because the low aqueous solubility of compound, it is unfavorable for the formation of preparation, it is necessary to configure cosolvent such as surfactant etc..Furthermore, poor water solubility can make these compounds to crystallize precipitation from solvent in preparation storage and/or transportation, so that the security for causing clinical drug to be applied has hidden danger.
In summary, find inhibited for PI3K and/or mTOR, and activity is good, selectivity is high and can solve the problem of dissolubility of such compound is low, effectively overcome the difficulty that the various preparations such as oral, intravenous injection, intramuscular injection are made, and expand the compound of clinical practice, it has also become the focus of current antineoplastic medicine research.The content of the invention
The applying date, such compound activity was good, selectivity is high, of the invention by the formula described in PCT/CN2013/001061 to disclose PI3K and/or mTOR inhibiting compounds in September, the 2013 International Application Serial No. PCT/CN2013/001061 of 12 days（I) compound is prepared into prodrug, the prodrug compound has the physicochemical properties for improving active compound, improve the selectivity that medicine is acted on target site, improve the pharmacokinetics processes such as absorption, distribution, transhipment and the metabolism of medicine in vivo, for the exploitation of preparation, the exploitation of crystal formation is significant.
It is therefore an object of the present invention to the prodrug of a class PI3K and/or mTOR inhibitors is provided, In particular it relates to：
(1) formula（I compound and its pharmaceutically acceptable salt and their stereoisomer shown in)：
A and B are separately CR6, R6For hydrogen, halogen atom, cyano group, hydroxyl, carboxyl ,-(CH2)nNR8aR8b、 -(CH2)nC(0)R9、 -(CH2)nC(0)NR8aR8b、 -(CH2)nOC(0)R9、 -(CH2)nC(0)(CH2)nOR9、 -(CH2)nN(R8a)C(0)R9, or optionally by the 1-3 C replaced selected from halogen atom, hydroxyl, carboxyl1-6Alkyl, alkoxy；
R1For hydrogen, or optionally by 1-5 R7aSubstituted C1-6Alkyl, C2-8Alkenyl, C2-8Block base, C3-8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles base, 7-12 member loop coils base, 7-12 member bridged ring bases；
R2For hydrogen, or optionally by 1-5 R7bSubstituted C1-6Alkyl, C2-8Alkenyl, C2-8Alkynyl, C3-8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles base, 7-12 member loop coils base, 7-12 member bridged ring bases；
R3For hydrogen, or optionally by the 1-3 alkyl replaced selected from halogen atom, hydroxyl, carboxyl； R4、 R5It is separately hydrogen, or optionally by the 1-3 d. replaced selected from halogen atom, hydroxyl, carboxyl6Alkyl；
M is 1 ~ 3；
E is hydrogen, or for can be with phosphoric acid forming salt inorganic base or the cation of organic base；
R7a、 R7bIt is separately
(1) halogen atom, cyano group, hydroxyl, trifluoromethyl ,-(CH2)nNR8aR8b、-(CH2)nC(0)R9、 -(C¾)nSR9、 -(CH2)nS(0)2R9、 -(CH2)nS (0) Makoto8aR8b、 -(CH2)nN(R8a)S(0)2R9、 -(CH2)nC(0)NR8aR8b 、 -(CH2)nOC(0)R9 、 -(CH2)nC(0)(CH2)nOR9 、 -(CH2)nN(R8a)C(0)R9,
(2) optionally by the 1-3 d_ replaced selected from halogen atom, hydroxyl, cyano group, trifluoro Yue bases6Alkyl, C2-8Alkenyl, C2-8Alkynyl, alkoxy；
(3) optionally by 1-3 selected from halogen atom, hydroxyl, trifluoromethyl, C alkyl, C2-8Alkenyl, C2-8Alkynyl, d-6 alkoxies ,-(CH2)nNR8aR8b、 -(CH2)nC(0)R9、 -(CH2)nSR9、 -(CH2)nS(0)2Ry、 -(CH2)nS(0)2NR8aR8b, -(CH2)nN(R8a)S(0)2Rv, -(CH2)nC(0)N 8aR8b 、 -(CH2)nOC(0)R9 、 -(CH2)nC(0)(CH2)nOR9 、 -(CH2)nN(R8a)C(0)R9Substituted C3-8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles bases；
R8a、 R8bSeparately be hydrogen, or optionally by 1-3 hydroxyl, halogen atom, cyano group, carboxyl,（This group is to use R8a、 R8bDefine R8a、 R8b) ^ sulfonyls, carbamoyl, sulfoamido substitution alkyl,.3.8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles bases；
R9For hydrogen, or optionally it is selected from halogen atom, cyano group, hydroxyl, carboxyl ,-(CH by 1-32)nNR8aR8b, sulfonyl, ^ Yue acyl groups substitution C1-6Alkyl, C1-6Alkoxy；N is 0 ~ 4.
(2) it is above-mentioned（1) formula described in（I compound and its pharmaceutically acceptable salt and their stereoisomer shown in), wherein：
A and B are separately CR6, R6For hydrogen, or optionally by the 1-3 d. replaced selected from halogen atom, hydroxyl, carboxyl6Alkyl；
R1For optionally by 1-3 R7aSubstituted 6-10 members aryl, the single heteroaryl of 5-6 members, the thick heteroaryl of 9-10 members, the single heterocyclic radical of 5-6 members, 9-10 member condensed hetero ring bases；
R2For optionally by 1-3 R7bSubstituted 6-10 members aryl, the single heteroaryl of 5-6 members, the thick heteroaryl of 9-10 members, the single heterocyclic radical of 5-6 members, 9-10 member condensed hetero ring bases；
R3For hydrogen；
R4、 R5It is separately hydrogen or d.6Alkyl；
M is 3;
E is hydrogen, or for can be with phosphoric acid forming salt inorganic base or the metal cation of organic base；
R7a、 R7bIt is separately
(1 ^ elements atom, cyano group, hydroxyl, trifluoro Yue bases ,-(CH2)nNR8aR8b、-(CH2)nC(0)R9、 -(CH2)nSR9、 -(CH2)nS(0)2R9、 -(CH2)nS(0)2NR8aR8b、 -(CH2)nN(R8a)S(0)2R9、 -(CH2)nC(0)NR8aR8b 、 -(CH2)nOC(0)R9 、 -(CH2)nC(0)(CH2)nOR9 、 -(CH2)nN(R8a)C(0)R9,
(2) optionally by the 1-3 C replaced selected from halogen atom, hydroxyl, cyano group, trifluoromethyl1-6Alkyl, alkoxy；
(3) halogen atom, hydroxyl, cyano group, trifluoromethyl, alkyl, alkoxy ,-(CH optionally are selected from by 1-32)nNR8aR8\ -(CH2)nC(0)R9、 -(CH2)nSR9、 -(CH2)nS(0)2R9、 -(CH2)nS(0)2NR8aR8b 、 -(CH2)nN(R8a)S(0)2Ry 、 -(CH2)nC(0)NR8aR8b 、 -(CH2)nOC(0)R9、 -(CH2)nC(0)(CH2)nOR9、 -(CH2)nN(R8a)C(0)R9Substituted C3.8Cycloalkyl, 5-10 unit's heteroaryls, 5-10 circle heterocycles bases；
R8a、 R8bIt is separately hydrogen or d_6 alkyl；
R9For hydrogen or alkyl；
N is 0 ~ 2.
(3) it is above-mentioned（2) formula described in（I compound and its pharmaceutically acceptable salt and their stereoisomer shown in), wherein：
A and B are separately CR6, R6For hydrogen or alkyl；
R4、 R5It is separately hydrogen；
M is 1;
E is hydrogen or sodium ion.
(4) it is above-mentioned（3) formula described in（I compound and its pharmaceutically acceptable salt and their stereoisomer shown in), wherein：
R1For optionally by 1-3 R7aSubstituted phenyl, the single heteroaryl of 5-6 members；
R2For optionally by 1-3 R7bSubstituted phenyl, the single heteroaryl of 5-6 members, the thick heteroaryl of 9-10 members；
(1) halogen atom, cyano group, hydroxyl, trifluoromethyl ,-NR8aR8b、 -C(0)R9、 -C(0)NR8aR8b、 -OC(0)R9、 -N(R8a)C(0)R9,
(2) optionally by the 1-3 C replaced selected from halogen atom, hydroxyl, cyano group, trifluoro Yue bases1-6Alkyl, C1-6Alkoxy,
(3) halogen atom, hydroxyl, cyano group, trifluoromethyl, alkyl, C_6 alkoxies ,-NR optionally are selected from by 1-38aR8b、 -C(0)R9、 -C(0)NR8aR8b、 -OC(0)R9、 -N(R8a)C(0)R9The substituted single heterocyclic radical of 5-6 members；
(1) Halogen elements atom, cyano group, hydroxyl, trifluoromethyl ,-NR8aR8b、 -C(0)R9、 -SR9、 -S(0)2R9、 -C(0)NR8aR8b、 -OC(0)R9、 -N(R8a)C(0)R9,
(2) optionally by 1-3 (^6 alkyl, the C alkoxies replaced selected from halogen atom, hydroxyl, cyano group, trifluoromethyl；
(3) optionally by 1-3 selected from halogen atom, hydroxyl, trifluoro Yue bases, alkyl, C alkoxies ,-NR8aR8b、 -C(0)R9、 -C(0)N 8aR8b、 -OC(0)R9、 -N(R8a)C(0)R9 The first single heteroaryls of substituted 5-6, the single heterocyclic radical of 5-6 members；
R8a、 R8bIt is separately hydrogen or d_6 alkyl；
R9For hydrogen or alkyl.
(5) it is above-mentioned（4) formula described in（I compound and its pharmaceutically acceptable salt and their stereoisomer shown in), wherein：
R1For optionally by 317&Substituted phenyl, pyrrole5It is ^, phonetic1^；
R2For optionally by 1-3 R7bSubstituted phenyl, pyridine radicals, pyrimidine radicals, thienyl, pyrazolyl, Yin11Base, Yin is sat to make an uproar base, pyrrole17Each simultaneously pyrrole of fixed and pyrrole radicals, pyrroleP^J^, pyrrole saliva and pyrrole>~, quinoline base；
R7aFor
(1) halogen atom, cyano group, hydroxyl, trifluoro Yue bases ,-NH2,
(2) optionally by the 1-3 C replaced selected from halogen atom, hydroxyl, cyano group, trifluoromethylMAlkyl,
(3) halogen atom, hydroxyl, cyano group, trifluoromethyl, C optionally are selected from by 1-31-4Alkyl, C4Piperidyl, the piperazinyl of alkoxy substitution；
R7bFor
(1) halogen atom, hydroxyl, trifluoromethyl ,-NH2、 -C(0)R9、 -SR9、 -S(0)2R9、 -NHC(0)R9,
(2) ^^, C are optionally cheated by the CM that 1-3 replace selected from halogen atom, hydroxyl, cyano group, trifluoromethyl>4 hole ^ ^;
(3) halogen atom, hydroxyl, cyano group, trifluoromethyl, C- optionally are selected from by 1-34Alkyl, Ci_4Alkoxy ,-NH2Substituted pyrrole radicals, pyrazolyl, imidazole radicals, piperidyl, piperazinyl, morpholinyl；
R9For hydrogen or d.4 alkyl.
(6) it is above-mentioned（5) formula described in（I compound and its pharmaceutically acceptable salt and their stereoisomer shown in), wherein：
R1For optionally by 1-3 173Substituted phenyl, pyrrole1^；
R2For optionally by 1-3 1715Substituted phenyl, pyridine radicals, pyrimidine radicals, thienyl, pyrazolyl, indazolyl, Yin tastes base, pyrrole and pyrrole radicals, pyrrolo- pyrrole " determine base, pyrazolo pyrrole ^, quinolyl；
(1) halogen atom, cyano group, hydroxyl, trifluoro Yue bases ,-NH2, (2) optionally by 1-2 methyl, ethyl, the isopropyls replaced selected from hydroxyl, cyano group, trifluoromethyl,
(3) optionally by 1-2 piperidyl, the piperazines replaced selected from hydroxyl, cyano group, trifluoro Yue bases, methyl, Yue epoxides.Qin Ji；
R7bFor
(1) Halogen elements atom, cyano group, hydroxyl, trifluoromethyl ,-NH2、 -SR9、 -S(0)2R9、 -NHC(0)R9,
(2) optionally by 1-2 Yue bases, ethyl, n-propyl, isopropyl, methoxyl group, the ethyoxyls replaced selected from hydroxyl, cyano group, trifluoromethyl；
(3) hydroxyl, cyano group, trifluoromethyl, methyl, ethyl, methoxyethoxy, _ NH optionally are selected from by 1-22Substituted pyrrole radicals, pyrazolyl, piperazinyl, morpholinyl;
R9For hydrogen, Yue bases or ethyl.
OOO/ OZXD/ works：) d OOO/ OZXD/ works：) d OOO/ OZXD/ works：) d
£^£000/M0ZN3/X3d Numbering Structure Number Structure
73 74 Yl 1
75
77
79
81
83
" halogen atom " of the present invention includes fluorine atom, chlorine atom, bromine atoms and iodine atom. " d_ of the present invention6Alkyl " refers to the straight or branched alkyl containing 1-6 carbon atom, and instantiation includes but is not limited to：Methyl, ethyl, n-propyl, isopropyl, normal-butyl, 2- methyl-propyls, 1- methyl-propyls, the Yue bases ethyls of 1,1- bis-, n-pentyl, 3- methyl butyls, 2- methyl butyls, 1- methyl butyls, 1- ethyl propyls, n-hexyl, 4- methyl amyls, 3- methyl amyls, 2- methyl amyls,
1- Yue bases amyl group, 3,3- dimethylbutyls, the Yue bases butyl of 2,2- bis-, 1,1- dimethylbutyls, 1,2- dimethylbutyls, 1,3- dimethylbutyls, 2,3- dimethylbutyls, 2- ethyl-butyls, 1,2- dimethyl propyls etc..
" C of the present invention3-8Cycloalkyl " refers to the cyclic alkyl containing 3-8 carbon atom, and instantiation includes but is not limited to：Cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, suberyl, cyclooctyl etc..
" C of the present invention2-8Alkenyl " can be straight or branched, including such as " C2_ 6 alkenyls ", " C2_ 4 alkenyls ", " C2-3Alkenyl ", " C3_ 6 cycloalkenyl groups " etc., instantiation includes but is not limited to：Vinyl, 1- Bing Women bases, 2- acrylic, 1- cyclobutenyls, 2- cyclobutenyls, 1,3-butadiene, 1- pentenyls,
2- pentenyls, 3- pentenyls, 1,3- pentadienes, 1,4- pentadienes, 1- hexenyls, 2- hexenyls, 3- hexenyls, 1,4- hexadienes, cyclopentenyl, 1,3- cyclopentadienyl groups, cyclohexenyl group, 1,4- cyclohexadienyls, cycloheptenyl, 1,4- cycloheptadiene base, cyclo-octene base, 1,5- cyclo-octadiene bases etc..（Seem be that the group on double bond both sides is cis and trans, it is unnecessary that this explanation seems advantage）
" C of the present invention2-8Alkynyl " can be straight or branched, including such as " Cw alkynyls ", " C2-4Alkynyl ", " C2-3Alkynyl ", " C3-8Block base " etc., instantiation includes but is not limited to：Acetenyl, propinyl, 2- butynyls, valerylene base, 3- pentynyls, 4- methyl-valerylene base, 2- hexin bases, 3- hexin bases, 5- methyl -2- hexin bases, 2- heptynyls, 5- methyl -2- heptynyls, 2- octynyls, 3- octynyls etc..
" CM alkoxies " of the present invention refers to " CW alkyl -0- ", wherein " Cw alkyl " text as defined above is described.
" 6-14 members aryl " of the present invention includes 6 ~ 8 unit monocycle aryl and 8 ~ 14 yuan of fused ring aryls.6 ~ 8 unit monocycle aryl are included such as phenyl, cyclooctatetraenyl.8 ~ 14 yuan of fused ring aryls include naphthalene, phenanthrene, 2,3- dihydros indenyl, indenyl, 1,2,3,4- tetralyls, 1,4- ihydro naphthyls etc.（It is recommended that：Aryl has had known implication, is tried not again for the suggestion of such group self-defined, to reduce error and save length so as to reduce cost）.
" 5-14 unit's heteroaryls " of the present invention includes the single heteroaryl of 5-8 members, the thick heteroaryl of 6-14 members, and described hetero atom has nitrogen, oxygen and sulphur etc., while including carbon atom, nitrogen-atoms and sulphur atom by the situation of oxo.
The instantiation of " the single heteroaryl of 5-8 members " includes but are not limited to furyl, thiophene p points of bases, pyrrole radicals, thiazolyl, isothiazolyl, the high base, isoxazolyls, oxadiazolyls of thiadiazolyl group, Evil, miaows Oxazolyl, pyrazolyl, 1,2,3- triazolyls, 1,2,4- triazolyls, 1 are the high bases of 2,3- Evil bis-, 1,2,4- oxadiazolyls, 1,2,5- oxadiazolyls, 1,3,4- oxadiazolyls, pyridine radicals, 2- pyridones, 4- pyridones, phonetic1^, 1, 4- Dioxin bases, 2-1, 2- oxazinyls, 4/ -1, 2- Evil bases, 6/ -1, 2- oxazinyls, 4/ -1, 3- oxazinyls, 6/ -1, 3- oxazinyls, 4/- 1, 4- oxazinyls, pyridazinyl, pyrazinyl, 1, 2, 3- triazine radicals, 1, 2, 4- triazine radicals, 1, 3, 5- triazine radicals, 1, 3, 4- triazine radicals, 1, 2, 4, 5- tetrazine bases, oxepin base, thin Xi base in heterocycle heptan three, azepine cycloheptatriene base, 1, 3- diaza cycloheptatriene bases, azepine cyclooctatetraenyl, 1, 4- dihydros -1, 4- diazacyclo sarohornenes, 1, 4- dioxane sarohornenes etc., preferably " the single heteroaryl of 5-6 members ".
The instantiation of " the thick heteroaryl of 6-14 members " includes but is not limited to：Benzofuranyl, benzisoxa furyl, benzothienyl, indyl, iso-indoles, benzoxazolyl, benzimidazolyl, indazolyl, BTA base, quinolyl, 2- quinolinones, 4- quinolinones, isoquinolines, isoquinolyl, acridinyl, phenanthridinyl, benzo pyridazinyl, phthalazinyl, quinazolyl, quinoxalinyl, phenol piperazine base, pteridine radicals, purine radicals, naphthyridines base, azophenlyene, phenthazine etc., are preferably " the thick heteroaryl of 9-10 members ".
" 5-10 unit's heteroaryls " of the present invention includes bicyclic heteroaryl and fused ring heteroaryl, and described hetero atom has nitrogen, oxygen and sulphur etc., while including carbon atom, nitrogen-atoms and sulphur atom by the situation of oxo.
" 3-14 circle heterocycles base " of the present invention includes the single heterocyclic radical of 3-8 members, 6-14 member condensed hetero ring bases.Described hetero atom has nitrogen, oxygen and sulphur etc., while including carbon atom, nitrogen-atoms and sulphur atom by the situation of oxo.
The instantiation of " the single heterocyclic radical of 3-8 members " of the present invention is included but are not limited to：Aziridine base, 2/7- aziridine base, diazacyclo propyl, 3-diazacyclo acrylic, azetidinyl, 1,2- diazetidines base, diazete base, 1,2- dioxetanes pit foundation,
1.2- diazetines base, dioxirane base, oxetanyl, oxaza propyl, Isosorbide-5-Nitrae-dioxane base, 1,3- dioxanes base, thiirane base, Thietane base,
1.3- dithian bases, 1,2- dithia cyclobutane bases, 1,3- dioxolane base, 1,3- dithiolane bases, 1,4- Dioxin bases, Isosorbide-5-Nitrae-dithiins base, Isosorbide-5-Nitrae-oxathiin base, tetrahydrofuran base, pyrrolin base, p ratios cough up alkyl, imidazolidinyl, 4,5- glyoxalidine bases, pyrazolidinyl, 4,5- pyrazoline bases, 2,5- dihydro fen bases, tetrahydro-thienyl, 4,5- dihydro-thiazolyls, piperidyl, piperazinyl, morpholinyl base, 4,5- dihydro-oxazole bases, 4,5- bis- Qing Yi Evil oxazolyls, 2,3- dihydro-isoxazole bases, 2-1,2- oxazinyls, 4/ -1,2- oxazinyls, 6/ -1,2- oxazinyls, 2-1,3- oxazinyls, 4-1,3- oxazinyls, 5,6- dihydro -4/7-1,3- oxazinyls, 6-1,3- oxazinyls, 2/- 1,4- oxazinyls, 4-1,4- oxazinyl, 2//- 1,3- thiazinyls, 4/ -1,3- thiazinyl, -1,3- the thiazinyls of 5,6- dihydros -4/, 6-1,3- thiazinyl, 2-1,4- thiazinyls, 4-1,4- thiazinyls, 2/- pyrans Base, 2-pyran-2-one base ,-2-pyranose of 3,4- dihydros, 4/- pyranose, THP trtrahydropyranyl, H- pyrans-4- ketone groups etc., are preferably " the single heterocyclic radical of 5-6 members ".
The instantiation of " 6-14 member condensed hetero rings base " of the present invention is included but are not limited to：Imidazolidine simultaneously [4, 5-c] pyridine radicals, 3, 4- dihydroquinazoline bases, 1, 2- dihydro-quinoxaline bases, benzo [[1, 3] dioxa cyclopentenyl, 1, 3- dihydroisobenzofuran bases, 2-chromogen alkenyl, 2//- chromogen alkene -2- ketone groups, 4-chromene base, 4//- chromene -4- ketone groups, Chromanyl, 4-1, 3- benzoxazinyls, 4, 6- dihydrofuran simultaneously [3, 4-] imidazole radicals, 3 α, 4, 6, 6-tetrahydrofuran simultaneously [3, 4- imidazole radicals, 4, 6- dihydro-thiophenes simultaneously [3, 4-t] imidazole radicals, 4, 6- pyrrolin simultaneously [3, 4- imidazole radicals, 4, 5, 6, 7- tetrahydro benzos [i/j imidazole radicals etc., preferably " 9-10 member condensed hetero rings base ".
" 5-10 circle heterocycles base " of the present invention includes single heterocyclic radical and condensed hetero ring base, and described hetero atom has nitrogen, oxygen and sulphur etc., while including carbon atom, nitrogen-atoms and sulphur atom by the situation of oxo.
" 7-12 member bridged rings base " of the present invention refers to that what any two ring shared two non-conterminous atoms formation contains 7-12 carbon atom or/and heteroatomic structure, described hetero atom has nitrogen, oxygen and sulphur etc., including such as " 7-10 members bridged ring ", " 7-9 members bridged ring ", " 7-8 members bridged ring ", " 7-8 members bridged ring ".The example includes but is not limited to for example：
Φ、 Θ、 、 、 ¾ gr、 、 、 What the term " 7-12 member loop coils base " of the present invention referred to the shared atom formation of at least two rings contains 7-12 carbon atom or/and heteroatomic structure, described hetero atom has nitrogen, oxygen and sulphur etc., including such as " 7-10 members loop coil ", " 7-9 members loop coil ", " 7-8 members loop coil ", " 7-8 members loop coil ".The example is included but are not limited to for example：
The compound of the present invention can be synthesized using the method described in following flows and/or other technologies known to persons of ordinary skill in the art, but be not limited only to following methods.
Intermediate 8
Reactions steps: (1) preparation of intermediate 2
By intermediate 1, (its preparation method is referring to WO2013/2071698 specifications page 38）, 1.5 times of equivalents R1 - NH2It is dissolved in appropriate potassium carbonate in the tert-butyl alcohol, heating response.Determine that after completion of the reaction, natural cooling, revolving removes solvent, is separated or recrystallized to obtain intermediate 2 by silica gel column chromatography with TLC monitorings.
(2) preparation of intermediate 3
Intermediate 2 is dissolved in energy appropriate solvent (such as methanol, ethanol or tetrahydrofuran) miscible with water, the aqueous solution of the lithium hydroxide of 3 times of equivalents is added dropwise.After completion of dropping, react 4 hours at room temperature.After reaction terminates, revolving removes solvent, plus suitable quantity of water, is separated out completely with salt acid for adjusting pH value to product, intermediate 3 is purified to obtain by suction filtration or recrystallization or column chromatography.
(3) preparation of intermediate 4
Intermediate 3 is suspended in appropriate thionyl chloride and reacted a few hours, concentration removes volatile materials.Then it is scattered in appropriate tetrahydrofuran, control temperature is 0.Under C, appropriate triethylamine is added dropwise and the amine shield (PG-NH of protection group is connected with2) mixture.Reaction solution is stirred at room temperature to by TLC monitoring reactions and finished, revolving removes solvent, passes through recrystallization or column chromatography obtains intermediate 4.
(4) preparation of intermediate 5
Intermediate 4 is scattered in appropriate ethyl chloroformate, is stirred at reflux, determines that reaction is finished to by TLC monitorings, revolving removes volatile materials, intermediate 5 is obtained by recrystallization or column chromatography.
(5) preparation of intermediate 6 selects corresponding reaction condition to slough amino protecting group, obtains intermediate 6 according to protection group PG difference.
(6) formula（Γ) the preparation of compound
Raw material 2 and appropriate organic or inorganic alkali, and palladium reagent and/or corresponding Phosphine ligands are placed in organic solvent（Such as Yue benzene, dioxane, dimethyl Yue acid amides, glycol dimethyl ether etc.) and water in the mixed solvent, under nitrogen protection, heating response to TLC monitors consumption of raw materials and finished, and formula is obtained by column chromatography for separation（Γ) compound.
(7) preparation of intermediate 7
By formula（Γ) raw material 3 of compound and 1.5 equivalents is dissolved in two Yue sulfoxides, sequentially adds potassium carbonate and sodium iodide, be stirred and heated to about 50 °C reaction 16 hours after, it is cooled to room temperature, then reaction solution is poured into water, filtered, dry intermediate 7.
(8) preparation of intermediate 8
Intermediate 7 is dissolved in dichloromethane, trifluoroacetic acid is added, reaction 3 hours is stirred at room temperature, Concentration, separates dry intermediate 8.
(9) formula（I) the preparation of compound
Intermediate 8 is dissolved in methanol, alkali, stirring reaction 10 minutes is added, revolving removes solvent and dry formula（I) compound.
In reaction equation, R R2、 R3、 R4、 R5, A, B, E and m as defined hereinabove, Hal Dai Biao Halogen element, selected from fluorine atom, chlorine atom, bromine atoms, iodine atom, preferably chlorine atom.
Formula（I) " pharmaceutically acceptable salt " of compound, refers to formula（I) acidic functionality present in compound（For example ,-COOH ,-OH, S03H etc.）With the salt of the cation formation of appropriate inorganic base or organic base, such as alkali metal salt；And formula（I) basic functionality present in compound（For example ,-NH2Deng）With appropriate inorganic acid or the salt of organic anion formation.（E is hereinbefore had been defined for cation, rather than salt）.
Formula（I) " stereoisomer " of compound, refers to work as formula（I when) there is asymmetric carbon atom, carbon-carbon double bond etc. in compound, produced all stereoisomers, including enantiomter, diastereoisomer, racemization isomers, cis-trans-isomer, dynamic isomer, geometric isomer, epimer and its mixture, it is included in the scope of the invention.
Formula of the present invention（I) compound, its pharmaceutically acceptable salt and their stereoisomer, can by oral administration, parenteral（Intravenous, intramuscular, subcutaneous or rectum etc.）, the administering mode such as transpulmonary, local be applied to mammal, such as people.The daily dose of the compounds of this invention can be about 5mg 500mg, preferably 50 ~ 300mg.
Formula（I) compound, its pharmaceutically acceptable salt and their stereoisomer, pharmaceutical composition can be made with one or more pharmaceutical carriers, pharmaceutically acceptable any formulation can be made, being applied in modes such as oral, parenterals needs the patient of this treatment.
, can be by formula during for being administered orally（I) compound, its pharmaceutically acceptable salt and their stereoisomer, conventional solid pharmaceutical preparation, such as tablet, glue Nang agent, pill, granule are mixed and made into conventional filler, adhesive, disintegrant, lubricant and/or diluent etc.；It may be made as oral liquid, such as mouth I solutions, the good supensoid agents of mouth Λ, syrup；
During for parenteral, injection, including parenteral solution, injection sterile powder and concentrated solution for injection can be configured to.
Formula（I) compound, its pharmaceutically acceptable salt and their stereoisomer can be used in treating and/or preventing proliferative diseases, it is possible to one or more other treatments Agent particularly antitumor agent and immunodepressant drug combination.The antitumor agent and immunodepressant are selected from antimetabolite, include but are not limited to capecitabine, gemcitabine, pemetrexed disodium；Growth factor receptor inhibitors, include but are not limited to pazopanib, Imatinib, Erlotinib, Lapatinib, Gefitinib, ZD6474；Antibody, includes but are not limited to Trastuzumab, bevacizumab；Mitotic inhibitor, includes but are not limited to taxol, vinorelbine, docetaxel, Doxorubicin；Antitumor steroids, includes but are not limited to Letrozole, tamoxifen, fulvestrant, Flutamide, Triptorelin；Alkylating agents, include but are not limited to endoxan, mustargen, melphalan, chlorambucil, BCNU；Metal platinum class, includes but are not limited to carboplatin, cis-platinum, oxaliplatin；Topoisomerase inhibitors, include but are not limited to topotecan, camptothecine, topotecan, Irinotecan；Immunosupress class, includes but are not limited to everolimus, sirolimus, special cancer and fits；Purine analogue, includes but are not limited to Ismipur, 6- thioguanines, stone and fills azoles purine；Antibioticses, include but are not limited to rhzomorph D, daunorubicin, adriamycin, mitoxantrone, bleomycin, plicamycin；Platinum complex, includes but are not limited to cis-platinum, Kapo Platinum；The upper gland cortex inhibitor classes of Kidney, include but are not limited to aminoglutethimide etc..Each medicine can be administered simultaneously or sequentially separate medication successively during drug combination, be administered with same dosage form or in the form of separated different preparations.
The invention further relates to formula U) purposes of compound, its pharmaceutically acceptable salt and their stereoisomer in the medicine for preparing treatment and/or prevention proliferative diseases such as tumour.
Described proliferative diseases include cancer and non-cancerous proliferative diseases, the cancer is selected from brain tumor, lung cancer, squamous cell, carcinoma of urinary bladder, stomach cancer, oophoroma, peritoneal cancer, cancer of pancreas, breast cancer, head and neck cancer, cervix cancer, carcinoma of endometrium, the carcinoma of the rectum, liver cancer, kidney, adenocarcinoma of esophagus, esophageal squamous cell carcinoma, solid tumor, prostate cancer, thyroid cancer, carcinoma in situ, lymthoma, neurofibromatosis, osteocarcinoma, cutaneum carcinoma, colon cancer, carcinoma of testis, gastrointestinal stromal tumor, mast cell tumor, Huppert's disease, melanoma, glioma or sarcoma etc.；The non-cancerous proliferative diseases are selected from such as the hyperplasia of prostate of skin or prostate.
It is demonstrated experimentally that the compounds of this invention is PI3K and/or mTOR inhibitors, to the proliferative diseases as caused by PDKa and/or mTOR signal path abnormal expressions for example（Malignant tumour) there is good therapeutic effect；And the compounds of this invention dissolubility is good, stable in physicochemical property, beneficial to the exploitation of ejection preparation, the internal pharmacokinetic studies of drug administration by injection are proved, the compound and with good pharmacokinetic properties, it is rapid-action, can be with drug administration by injection, the problem that critical patient can not be administered orally can be effectively solved, expands clinical practice.
The beneficial effect of the compounds of this invention set forth further below, other compounds of the invention and experiment In cited part the compounds of this invention there is identical beneficial effect, but this should not be interpreted as to the compounds of this invention only there is following beneficial effect.
Representative implication of being abridged in following experiments is as follows：
HEPES:The horizontal acid of hydroxyethyl piperazine second sulphur；
EGTA:Ethylene glycol diethyl ether ethylenediamine tetraacetic acid (EDTA)；
CHAPS:3- [the Yue amino of 3- (courage amido propyl) two] propane sulfonic acid inner salt
DTT:Two ^ L threitols；
PIP2:4,5- diphosphonic acid phosphatidylinositols；
ATP:Atriphos；
DMSO:Dimethyl sulfoxide (DMSO)；
Tween-20:Polysorbas20.The external zymetology inhibitory activity of compound A, B of experimental example 1
Compounds A, B are tested, gained is prepared by embodiment 1,3；
MTOR zymetology experimental methods
1. preparation of reagents
1.1 1 times of kinases Slow fliud flushings： 50 mM HEPES , pH 7.5 , 10 mM MgCl2, 1 mM EGTA, 3 mM MnCl2, 0.01 % Tween-20 , 2 mM DTT;
1.2 4 times of kinase solutions：MTOR kinases is added in 1 times of kinases Slow fliud flushing, 4 times of kinase solutions, final concentration of 2.5 nM are prepared;
1.3 2 times of substrates and ATP solution：Substrate 4EBP1 and ATP are added in 1 times of kinases Slow fliud flushing, 2 times of substrate solutions, the final concentration of 10.8 μ Μ of 4EBP1 final concentration of 50 nM, ATP are prepared;
1.4 4 times of tester solution：The tester solution of 100 times of various concentrations gradients is prepared using 100 % DMSO, 25 times is diluted with 1 times of kinases Slow fliud flushing, obtains the tester solution of 4 times of various concentrations gradients；
1.5 detect the preparation of liquid：Prepare the detection liquid containing 2 times of final concentration EDTA and 4EBP1 phospho-ABs, final concentration of 8 mM of EDTA, final concentration of 2 nM of 4EBP phospho-ABs.
2. reality ^^ steps
2.1 add 2.5 4 times of tester solution being serially diluted, multiple holes toward every hole in 384 orifice plates；
2.2 add 2.5 4 times of kinase solutions per hole, and vibrations are mixed； 2.3 add 52 times of substrates and ATP solution per hole, are incubated at room temperature 1 hour；
2.4 are eventually adding 10 detection liquid terminating reactions, after 60 minutes, and Envision reads data Lance signal (665 nM).
3. data processing
Inhibiting rate %=(sample value-minimum value)/(maximum-minimum value) χ 100
Wherein " maximum " is DMSO control wells readings, and " minimum value " is the control wells reading for being not added with kinases.
Input GraphPad Prism 5,0 to map, obtain curve and IC50。
The Κ α zymetology experimental methods of Ρ Ι 3
1. preparation of reagents
1.1 1 times of kinases Slow fliud flushings：50 7.5,3 mM MgCl of mM HEPES, pH2, 1 mM EGTA, 100 mM NaCl, 0.03 % CHAPS, 2 mM DTT;
1.2 4 times of kinase solutions：The Κ alpha kinases of Ρ Ι 3 are added in 1 times of kinases Slow fliud flushing, 4 times of kinase solutions, final concentration of 1.65 nM are prepared;
1.3 2 times of substrates and ATP solution：Substrate PIP2 and ATP are added in 1 times of kinases Slow fliud flushing, 2 times of substrate solutions, the final concentration of final concentration of 25 μ Μ of 50 μ Μ, ATP of PIP2 are prepared;
1.4 4 times of tester solution：The tester solution of 100 times of various concentrations gradients is prepared using 100 % DMSO, 25 times is diluted with 1 times of kinases Slow fliud flushing, obtains the tester solution of 4 times of various concentrations gradients；
1.5 K ina s e- G l o reagent reagents, are put to room temperature is warming up to, and for terminating reaction and produce detection signal.
2. experimental procedure
2.1 add 2.5 4 times of tester solution being serially diluted toward every hole in 384 orifice plates；
2.2 add 2.5 4 times of kinase solutions per hole, and vibrations are mixed；
2.3 add 5 L, 2 times of substrates and ATP solution per hole, are incubated at room temperature 1 hour；
2.4 are eventually adding 10 μ L detection solution terminating reactions, and Slow was shaked after 15 minutes slowly, and Flexstation reads data RLU.
3. data processing
Inhibiting rate./.=100-(sample value-minimum value)/(maximum-minimum value）χ 100
Wherein " maximum " is the control wells reading for being not added with kinases, and " minimum value " is DMSO control wells readings.
GraphPad Prism5.0 mappings are inputted, curve and IC is obtained50。 Experimental result
External the enzyme activity (the IC of compound A, B50)
Tester Ρ Β Κ α (η M) mTOR (η Μ)
Compound A 7.8 2.5
Compound B 15.2 2.66
Experiment conclusion
As can be seen from Table 1, compound Α, Β is respectively provided with good inhibitory activity to Κ α and the mTOR enzymes of Ρ Ι 3 in vitro, illustrates that compound A, B has inhibitory action to the disease as caused by Κ α and mTOR the signal path abnormal expressions of Ρ Ι 3.The compound A of experimental example 2 cell in vitro inhibitory activity
Compounds A is tested, gained is prepared by embodiment 1；
The implication that cell line used is represented in following experiments is as follows：
A549:Human lung cancer carefully wraps strain；
U87MG:Human brain astrocytes' blastoma cell line；
PC-3:Human Prostate Cancer Cells woods；
SKOV-3:Human oophoroma cell line；
Experimental method
1. reagent and compound are prepared
1.1 tenth of the twelve Earthly Branches, oneself put stone Lin acid Slow fliud flushings（PBS ):8 g NaCl, 0.2 g KCl, 1.44 g Na are weighed respectively2HP04With 0.24 g KH2P04, 800 mL ultra-pure waters are added, pH=7.4 are adjusted to, ultra-pure water is added, is settled to 1 L, the min of autoclaving 20.
1.2 configuration XTT detection working solutions：Weigh the nitrogen salt of lOO mg methyl four（XTT) powder, lucifuge, which is dissolved in 300 mL and is heated to 50 °C, to be free of in phenol red serum-free RPMI1640 nutrient solutions, is filtered, packing, immediately or one week it is interior use, all processes need lucifuge.
1.3 configuration testing compounds
Configuration testing Compound Stock solution：Compound powder is dissolved in DMSO, concentration is 10 mM.
Configuration testing compound gradient dilution solution：10 mM test compound liquid storage is taken to use
4 times of continuous gradient dilutions of DMS0, totally 10 concentration.Then the compound for taking 2 DMSO to dilute respectively is added to 998 μ containing 10% hyclone（FBS in nutrient solution), tester maximum concentration is that 20 μ Μ, DMSO concentration are 0.2 %, totally 10 concentration gradients. 2. cell culture
2.1 cell recovery
Cell cryopreservation tube is taken out from liquid nitrogen, is placed in 37 °C ~ 39 °C water-baths, fast melt.Frozen stock solution is transferred in 15 mL sterile centrifugation tubes, the nutrient solution of 10 times of frozen stock solution volumes is added, in 1000 rpm, 4 °C of 5 min of centrifugation.Nutrient solution in reject centrifuge tube, power mouth enters the nutrient solution containing 10% FBS, and cell is resuspended, is transferred in blake bottle, changes liquid within second day.
2.2 passage
Take the logarithm growth period cell, reject nutrient solution adds appropriate volume PBS and washed once, add the digestive juice containing 0.25 % pancreatin and 0.02 % EDTA of appropriate volume.37 °C of 2 ~ 5 min of placement, discard digestive juice, PBS is washed once.Add nutrient solution of the appropriate volume containing 10 % FBS and terminate digestion, pipette is gently blown and beaten, cell suspension will be made after cell dissociation for passage and test.
2.3 cell cryopreservation
Take the logarithm growth period cell, with the digestive juice vitellophag containing 0.25 % pancreatin and 0.02 % EDTA, cell suspension is made, in 1000 rpm, 4 °C of 5 min of centrifugation.Nutrient solution is discarded, the frozen stock solution containing 10 % DMSO and 90 % FBS is added, cell is resuspended, often the χ 10 of pipe 26Individual cell is sub-packed in cryopreservation tube.Cryopreservation tube is placed in program temperature reduction box, after -80 °C are placed 24 h, is transferred in liquid nitrogen and freezes.
3. plating cells
3.1 prepare cell suspension：Remove the culture medium in blake bottle；With twice of PBS rinses cell；Plus pancreatin digestion is collected by centrifugation；It is resuspended with the culture medium containing 10 % FBS hyclones, counts and be adjusted to suitable concn（90%) cell viability have to be larger than;Cell concentration is 5 X 104/mL;
Cell suspension is added 96 orifice plates per the μ of hole 100 by 3.2, is placed in 37 °C, 5 % C02Overnight incubation in cell culture incubator；
4. drug-treated：The test compound diluted is added into Tissue Culture Plate, totally three repetitions are 10 μ Μ per the initial concentration of 100 final volume of hole 200,4 times dilute, totally 10 concentration gradients；It is put into C02Cultivated 72 hours in cell culture incubator；
5. XTT methods detect cell viability：Culture medium is removed, XTT detection working solutions are added, 150 per hole, in 37 °C, 5 % C02Placed 2 hours in cell culture incubator, be put into ELIASA and read 450 nm extinctions；
6. data processing
1) inhibiting rate %=(solvent control hole reading-tester hole reading)/(solvent control hole reading White control wells reading) χ 100%;
2) input GraphPad Prism 5.0 map, and obtain curve and IC
Experimental result
Compound A cell in vitro activity (IC50)
Tester U87MG (nM) A549 (nM) PC-3 (nM) SKOV-3 (nM) compounds A 29.48 63.16 33.73 51.17
Experiment conclusion
Κ α and mTOR the signal path abnormal expressions of Ρ Ι 3 can cause the increment of U87MG, A549 cell; as can be seen from Table 2; compound A can effectively suppress the propagation of U87MG, A549, PC-3, SKOV-3 cell, illustrate that compound A has inhibitory action to the disease as caused by Κ α and mTOR the signal path abnormal expressions of Ρ Ι 3.The Pharmacokinetics in Rat experiment of the compounds of this invention 1 of experimental example 3
Animal subject male SD rat, 3/method of administration/compound, body weight 230-250 g.Test sample
The compounds of this invention 1, gained is prepared by embodiment 2, is dissolved with 30 %DMF+70 % sterilizeds water for injection.Catalysis or non-enzyme effect of the compounds of this invention 1 in rat body by enzyme, reactive compound A is converted into play pharmacological action, reactive compound A is referred to as the active compound of the compounds of this invention, thus LC-MS/MS monitoring analyses be compound A blood concentration.
Compound A, gained is prepared by embodiment 1, using 0 ° of 30%DMF+5/.(0. 9% sodium chloride injections of PEG400+2 0%）（Use hydrochloric acid water.
Compound A
Internal standard compound
BEZ-235:It is prepared by the compound disclosed in WO2006122806 (publication date 2006.11.23), the method with reference to described in this application.
Experimental method
The administration of test sample intravenous injection is administered（Iv), dosage is 2mg/kg, the mg/kg of administration concentration 1, the mL/kg of administered volume 2;Test sample gastric infusion（Po), dosage is 4 mg/kg, the mg/kg of administration concentration 1, the mL/kg of administered volume 4.
0.083,0.25,0.5,1 after blood sampling IV administrations, 2,4,6,8,24 h, carry out tail vein blood, 0.167,0.5 after PO administrations, 1,2,4,6,8,24 h, carry out tail vein blood, each time point takes 100 μ L or so whole blood to be placed in liquaemin anticoagulant tube, 6 min separated plasmas is centrifuged in 8000 revs/min of high speed low temperature centrifugal machine, blood plasma freezes in -80 °C of casees.
Plasma sample analysis
Test sample uses precipitation of protein：20 blood plasma are taken, 200 μ internal standards are added（The BEZ-235 ng/mL of methanol solution 50), then 1500 revs/min of 3 min of vortex centrifuge 5 min in 12000 revs/min of supercentrifuge, take supernatant 50 to add 150 water, be vortexed and mix.
Sample is followed the trail of
LC-MS/MS detects prodrug compound 1 and its active compound compound A blood concentration simultaneously.2 mg/kg are administered in the IV of prodrug compound 1, the mean blood plasma concentration of the prodrug compound 1 of 5 tri- animals of min is about 700 ng/mL, 15mm mean blood plasma concentrations are about 50 ng/mL, 30 min and later time point are not detected by blood concentration substantially, illustrate that prodrug compound 1 changes into active compound compound A within 30 min;
4 mg/kg are administered in the PO of prodrug compound 1, and all blood sampling time points are not detected by the blood concentration of prodrug compound 1, illustrate the fast speed in rat body of prodrug compound 1 substantially（Within 5 min）Change into active compound compound A. The P of Rats K evaluation results of the compounds of this invention（iv )
AUClast CI VssTest sample
(h*ng/mL) (L/h/kg) (L/kg) compound 1
(determine by compound 1 convert 8,687 0.23 0.85 one-tenth compound A data）
The P of Rats K evaluation results of the compounds of this invention of 9,130 0.21 1.26 tables of compound A 4（po )
T AUClast C max Tmax
Test sample F%
(h) (h*ng/mL) (ng/mL) (h)
Compound 1
(determine by compound 1 convert 9.04 1,695 143.3 4.00 12.2 one-tenth compound A data）
Compound A 5.77 2,549 205 6.00 14.0
AUCiasT represents area under the drug-time curve0→t
CL represents clearance rate
VssRepresent apparent steady state distribution volume
T1/2Represent half-life period
TmaxRepresent blood medicine peak time
cmaxRepresent blood peak concentration of drug
F% represents absolute bioavailability experiment conclusion
Prodrug compound 1 carries out rat IV and PO administration, and active compound compound A can be quickly completely converted into rat body.From upper table 3 and 4, the compounds of this invention 1 and compound A are respectively provided with good pharmacokinetic properties.After the administration of the rat of compound 1, the Pharmacokinetic Characteristics for monitoring compound A are similar with Pharmacokinetic Characteristics such as clearance rate, AUC and the bioavilability that compound A monomers are administered, illustrate that compound 1 can change into its active compound compound A in rat body, pharmacological activity is played by compound A, compound A is prepared into its prodrug compound 1, the pharmacological activity of compound in itself is not influenceed, illustrates that the compounds of this invention 1 has for the disease such as tumour as caused by PDKct and mTOR signal path abnormal expressions significant Inhibitory action.The compounds of this invention 1 of embodiment 4 and compound A solubility property compare
Test sample：The active compound compound A of the compounds of this invention 1, gained is prepared by embodiment 1.
The compounds of this invention 1, gained is prepared by embodiment 2；
The chromatographic condition of the compounds of this invention 1
Instrument：High performance liquid chromatography is discussed（ Agilent 1200 series )
Chromatographic column：Agilent Eclipse XDB-Cis (filler is 5 μ η ι octadecylsilane bonding stone Gui glue, the mm of internal diameter 4.6, the mm of column length 150)
Column temperature： 30 °C;Flow velocity： 1.0 mL/min;Detection wavelength： 214 nm;Sample size： 10 L;Mobile phase：
Slow fliud flushings：0.02 mol/L ammonium dihydrogen phosphates（0.2% triethylamine is added, and with phosphorus acid for adjusting pH value to 6.0)
Mobile phase A:Buffer solution-acetonitrile（90:10) Mobile phase B:Slow fliud flushings-acetonitrile（20:80) condition of gradient elution：
Time（Minute）Mobile phase A (%) Mobile phase B (%)
0 80 20
5 50 50
10 20 80
12 0 100
18 0 100
20 80 20
The 22 80 20 compound A general conditions of color i
Instrument：High performance liquid chromatography is discussed（ Agilent 1200 series )
Chromatographic column： YMC-Pack-Pro C18(filler is 5 μ π ι octadecylsilane chemically bonded silica, the mm of internal diameter 4.6, the mm of column length 150)
Column temperature： 30 °C ;Flow velocity： 1.0 mL/min;Detection wavelength： 214 nm;Sample size：10 μ ^ mobile phases：
Slow fliud flushings：0.02 mol/L ammonium dihydrogen phosphates（0.2% triethylamine is added, and with phosphorus acid for adjusting pH value to 6.0) Mobile phase A:Slow fliud flushings-acetonitrile（90:10) Mobile phase B:Slow fliud flushings-acetonitrile（20:80) mobile phase A:Mobile phase B=65:35
The solution of test sample is prepared：
The solution of the compounds of this invention 1 is prepared：
The solution of test sample the compounds of this invention 1：
Precision weighs two parts of 2 mg of test sample, puts in suitable vessel, is separately added into pH4.0, pH5.0 Slow fliud flushings 0.5 mL, ultrasonic 5 min, then is placed in 25 °C of water-baths 2 hours, filtering, takes filtrate as need testing solution.
Precision weighs the mg of test sample 2, puts in suitable vessel, adds pH6.0 Slow fliud flushings 0.5 mL, ultrasonic 5 min, and filtering takes filtrate as need testing solution.
Precision weighs two parts of 2 mg of test sample, puts in suitable vessel, is separately added into pH7.0, pH9.0 Slow and rushes solution in right amount, records the solvent volume added every time, shaking until being completely dissolved, be used as need testing solution.
The solution of reference substance the compounds of this invention 1：
Precision weighs the mg of reference substance 4.7, puts in 10 mL measuring bottles, and adding DMSO lmL dissolves it, and again with methanol is settled to scale and the solution for containing 0.47 mg in every 1 mL is made, and enters as mother liquor.With Yue alcohol, by mother liquor A, progressively reference substance B and reference substance containing 0.047 mg, 0.0094 mg is made in every 1 mL in dilution respectively（.
Compound A solution is prepared：
For test product Compound A solutions：
Precision weighs tri- parts of 2 mg of test sample, is separately added into pH5.0, pH7.0, pH9.0 Slow fliud flushings 2 mL, ultrasonic 5 min, filtering, takes filtrate as need testing solution.
Reference substance Compound A solution：
Precision weighs the mg of reference substance 12.47, plus the solution for containing 0.2494 mg in every 1 mL is made in the mL of DMSO 50, shakes hook, is used as reference substance mother liquor.Mother liquor A is progressively diluted respectively with DMSO again the solution for containing 0.04988 mg, 0.02494 mg, 0.009976 mg, 0.004988 mg, 0.002494 mg in every 1 mL is made, be used as each reference substance solution.
Result of the test：
With each reference substance concentration (C) for abscissa, peak area（Y) it is ordinate, carries out linear regression, obtain linear equation.Each need testing solution peak area substitution linear equation is drawn into need testing solution concentration.
The reference substance linear equation of the compounds of this invention 1 is：Y=18821C, coefficient correlation corpse 0.9999. Compound A/controls product linear equation is：Y=29144C, correlation coefficient r=1.0000.
Measurement result is as follows：
Solubility theoretical concentration (mg/mL) the Slow fliud flushing pH value measured concentrations (mg/mL) of the compound A of table 5 under different pH Slow fliud flushings
5 1.89x 1ο-5
Solubility of the compounds of this invention 1 of 90 table 6 under different pH Slow fliud flushings
Theoretical concentration (mg/mL) Slow fliud flushing pH value measured concentrations (mg/mL)
4 0.189
4 5 0.556
6 3.20
17 >=5 and< 7
19 >=15 and<20 experiment conclusions
From table 5 and table 6, when Slow fliud flushings pH value is 5,7,9, compound A dissolving measured concentration is respectively lower than the measured concentration of identical Slow fliud flushings pH value the compounds of this invention 1, and dissolubility of the dissolubility than compound A for illustrating the compounds of this invention 1 more preferably, there is significant difference；And the dissolubility of the compounds of this invention increases with the increase of Slow fliud flushing pH value, more preferably, beneficial to pharmaceutically acceptable any formulation is made, injection can be especially made in physicochemical property.Solubility of the compounds of this invention of embodiment 5 in water compares
Test sample：The active compound compound B of the compounds of this invention 14, gained is prepared by embodiment 3.
The compounds of this invention 14, gained is prepared by embodiment 4；The active compound compound C of the compounds of this invention 31, gained is prepared by embodiment 5.The compounds of this invention 31, gained is prepared by embodiment 6；Experimental method：Each 2 mg of compound 14, compound B, compound 31, compound C sample is taken respectively, gradually adds ultra-pure water, each ultrasonic 5 min, until sample is completely dissolved.
Experimental result：Solubility of the compound B in water< 0.1 mg/mL.
Solubility of the compound 14 in water> 5 mg/mL;
Solubility of the compound C in water is less than 0.1 mg/mL. Solubility of the compound 31 in water> 5 mg/mL;
Experiment conclusion：
It can be seen that by result above, solubility of the compound 14 in water is 50 times of solubility of its active compound compound B in water, solubility of the compound 31 in water is 50 times of solubility of its active compound compound C in water, it follows that, the prodrug compound physicochemical property of the present invention is more preferable, beneficial to pharmaceutically acceptable formulation is made, injection can be especially made, can effectively expand the exploitation of clinical application formulation.Embodiment
The embodiment of form, is described in further detail to the above of the invention by the following examples.But the scope that this should not be interpreted as to above-mentioned theme of the invention is only limitted to following examples.
EtOAc:Ethyl acetate；
EA:Ethyl acetate；
PE:Petroleum ether；
HATU:2- (7- azos BTA)-Ν, Ν, Ν ', the fluorine Brick acid esters of Ν '-tetramethylurea six； DCM:Dichloromethane；
DMF:Dimethylformamide.
5- (4,4,5,5- tetramethyls -1,3,2- dioxaborolanes -2- bases) pyridine -2- amine, self-control, preparation method is as follows：
By the bromopyridine of 2- amino -5 (5.1 g, 30 mmol), connection boric acid pinacol ester (10.7 g, 45 mmol), potassium carbonate（8.3 g, 60 mmol), four（Triphenylphosphine bar (693 mg, 0.6 mmol) is added in 150 mL dioxane and 2 mL water, under nitrogen protection back flow reaction 4 hours.Room temperature is cooled to, is filtered, concentration, crude product is dissolved in 300 mL dichloromethane, is washed with water, with anhydrous sodium sulfate drying, when being concentrated into remaining a small amount of solvent, petroleum ether is added thereto and separates out yellow solid, filtering obtains product（1.8 g ).The 9- of embodiment 1 (6- aminopyridine -3- bases) -1- (3- (trifluoromethyls）Phenyl）Pyrimido
[5,4-c】【L, 5】Naphthyridines-2,4 (1 3-diketone（Compound A) preparation
1) the chloro- 4- of 6- ((3- (trifluoromethyl) phenyl))-l, 5- naphthyridines -3- Yue acetoacetic esters
By the chloro- 1,5- naphthyridines -3- carboxylic acid, ethyl esters of 4,6- bis- (5.4 g, 20 mmol), (its preparation method is referring to WO2013/2071698 specifications page 38）, 3-Aminotrifluorotoluene (4.5 g, 28 mmol) and potassium carbonate (5.5 g, 40 mmol) be added in the 150 mL tert-butyl alcohols, be heated to 90 °C and react 18 hours.Reaction solution is cooled to after room temperature and is spin-dried for, add 300 mL water, gained solid is filtered, with ethyl acetate and petroleum ether (1/20,50 mL) washing, the chloro- 4- of 6- ((3- (trifluoromethyl) phenyl) amino) -1,5- naphthyridines -3- Ethyl formates are obtained, are yellow solid (6.0 g).
2) 6- chlorine
By the chloro- 4- of 6- ((3- (trifluoromethyl) phenyl) amino) -1,5- naphthyridines -3- Ethyl formates (3.95 g, 10 mmol) it is dissolved in 50 mL methanol and 50 mL tetrahydrofurans, water (50 mL) solution of lithium hydroxide (1.26 g, 30 mmol) is added dropwise.After completion of dropping, react 4 hours at room temperature.Reaction solution is concentrated, 200 mL water are added, with salt acid for adjusting pH value to 3, the filtering of gained solid, vacuum thousand is dry to obtain yellow solid (3.6 g：).
3) Λ 4- methoxybenzyls) the chloro- 4- of -6- ((3- (trifluoromethyl) phenyl) amino) -1,5- naphthyridines -3- Yue acid amides
By the chloro- 4- of 6- ((3- (trifluoromethyl) phenyl) amino) -1,5- naphthyridines -3- carboxylic acids (3.6 g, 9.8 mmol) it is suspended in 50 mL thionyl chlorides, it is heated with stirring to 75 ° and (keeps reaction 4 hours.Room temperature is naturally cooled to, concentrates, obtains yellow solid.It is scattered in 100 mL tetrahydrofurans, the mixture of triethylamine (3.03 g, 30 mmol) and 4-Methoxybenzylamine (1.6 g, 13 mmol) is instilled under 0 °C.Reaction solution is stirred at room temperature 4 hours, and revolving removes solvent, and 300 mL water are added to residue, Suction filtration, filter cake ethyl acetate and petroleum ether (volume ratio 1/10,100 mL) washing, dry, obtain Λ 4- Yue oxygen benzyl) the chloro- 4- of -6- ((3- (trifluoromethyl) phenyl) amino) -1,5- naphthyridines -3- formamides, are faint yellow solid (4.5 g).
4) the chloro- 3- of 9- (4- methoxy-benzyls)-1-(3- (trifluoromethyl) phenyl) pyrimido [5, 4-c] [1,5] naphthyridines -2,4 (1/, 3)-diketone preparation
By Yue oxygen benzyl) the chloro- 4- of -6- ((3- (trifluoromethyl) phenyl) amino) -1,5- naphthyridines -3- formamides (4.5 g, 9.2 mmol) it is suspended in ethyl chloroformate (50 mL), 90 V are heated to, are stirred 120 hours.Revolving removes volatile materials, silica gel column chromatography (EtOAc/PE=0 l/4) isolated 9- chlorine _ 3- (4- Yue oxy-benzyls）- 1-(3- (trifluoromethyls）Phenyl）Pyrimido [5,4-c] [l, 5] naphthyridines -2,4 (1/, 3 /)-diketone, is yellow oil (700 mg).
5) small (3- (trifluoromethyl) phenyl) pyrimido [5,4- of 9- chlorineC] [l, 5] naphthyridines -2,4 (1/, 3 /)-diketone preparation
By the chloro- 3- of 9- (4- methoxy-benzyls) -1- (3- (trifluoromethyl) phenyl) pyrimido [5,4-c] [l, 5] naphthyridines -2,4 (1,3)-diketone (700 mg, 1.36 mmol) is dissolved in acetonitrile (40 mL) and water（10 mL) in, ammonium ceric nitrate (2.9 g, 5.65 mmol) is added portionwise at room temperature.Reaction solution is stirred at room temperature after 18 hours revolving and removes solvent, crude product silica gel column chromatography (EtOAc/PE=0 l/4) isolated yellow solid (400 mg).
6) small (3- (trifluoro Yue yls) phenyl) pyrimido [5,4-c] [l, the 5] naphthyridines -2,4 of 9- (6- aminopyridine -3- bases) (1 //, 3/)-diketone
By 9- chloro- 1-(3- (trifluoromethyl) phenyl) pyrimido [5, 4-c] [1, 5] naphthyridines -2, 4 (1, 3/)-diketone (400 mg, 1 mmol), 5- (4, 4, 5, the Yue bases -1 of 5- tetra-, 3, 2- dioxaborolanes -2- bases) pyridine -2- amine (440 mg, 2.0 mmol), potassium carbonate (414 mg, 0111101) and tetrakis triphenylphosphine palladium (58 mg 3.0, 0.05 mmol) it is added in 40 mL dioxane and 2 mL water, the h of back flow reaction 18 under nitrogen protection, it is cooled to room temperature, diatomite is filtered, concentration, silica gel column chromatography (EtOAc/PE=0 ~ 10/l) isolated crude product, washed with Yue alcohol, obtain 9- (6- aminopyridine -3- bases) -1- (3- (trifluoro Yue yls) phenyl) pyrimido [5, 4-c] [l, 5] naphthyridines -2, 4 (1, 3 /)-diketone, for yellow solid (56 mg).
Molecular formula： C22H13F3N602 LC-MS(m/e): 451.1(M+H)
^-NMR (400 MHz, DMSO-^6) δ:12.28 (br. s., IH), 9.26 (s, IH), 8.33 (m, IH), 8.21-8.26 (m, IH), 8.16 (m, IH), 7.93 (s, 1H), 7.81 (br. s., 1H), 7.70 (m, 2H), 6.76 (m, 1H), 6.46 (s, 2H), 6.24 (m, IH the) (9- (6- aminopyridine-3- bases)-2,4- dioxos-l- (3- (trifluoromethyls of embodiments 2）Phenyl) -1,2- dihydro-pyrimidins simultaneously [5,4-c】[l, the preparation of (4-yl) the methyl phosphorodithioate disodium (compound 1) of 5 " naphthyridines-3
1) (9- (- 3-yl of 6- aminopyridines)-2,4- dioxos-1-(3- (trifluoro Yue yls) phenyl)-1,2- dihydro-pyrimidins simultaneously [5,4-c | [l, 5] naphthyridines-3 (4/)-yl) methyl di-t-butyl phosphate preparation（After think that naphthyridines is better, be also beneficial in the later stage translate English, so benzodiazine is uniformly changed into naphthyridines）
By small (3- (trifluoromethyl) phenyl) pyrimidos [5 of 9- (6- aminopyridine -3- bases), 4-c] [l, 5] naphthyridines -2,4 (1,3/diketone (900mg, 2mmol) (compound A) and di-t-butyl chloromethyl phosphate (are purchased from Nanjing Jing Ruijiuan Bioisystech Co., Ltd）(673 mg, 2.6 mmol) are dissolved in 15 mL DMSO, sequentially add K2C03(673 mg, 2.6 mmol) and Nal (30 mg, 0.2 mmol), be stirred and heated to 50 °C keep 16 hours after, be cooled to room temperature, then reaction solution be poured into water, suction filtration, filtration cakes torrefaction obtains the g of crude title compound 1.5.
2) (small (3- (trifluoromethyl) phenyl) phonetic-c of -1,2- dihydros of 9- (6- aminopyridine -3- bases) -2,4- dioxos] [l, 5] naphthyridines -3 (4)-base）The preparation of methyl dihydrogen phosphoric acid ester
By (9- (6- aminopyridine-3- bases)-2,4- dioxos-1-(3- (trifluoromethyl) phenyl)-1,2- dihydro-pyrimidins simultaneously [5,4-c] [l, 5] naphthyridines-3 (4/ /)-yl) methyl di-t-butyl phosphate (1,5 g crude products）It is placed in 10 mL dichloromethane, adds trifluoroacetic acid（10 mL), it is stirred at room temperature 3 hours, is concentrated in vacuo, C-18 ODS chromatographic columns（Methanol/water, 0 ~ 60%) isolated faint yellow solid, is washed with a small amount of methanol, dry title compound (150 mg).
3) (9- (6- aminopyridine -3- bases) -2,4- dioxos -3- (trifluoromethyl) phenyl) -1,2- dihydro-pyrimidins [5,4-c] [l, 5] naphthyridines -3 (4/ /)-yl) methyl phosphorodithioate disodium salt preparation
By (9- (6- aminopyridine -3- bases) -2,4- dioxos are small (3- (trifluoromethyl) phenyl) -1,2- dihydro-pyrimidins simultaneously [5,4-c] [l, 5] naphthyridines -3 (4)-yl) Yue base dihydrogen phosphoric acids ester (150 mg, 0.27 mmol) is placed in In 10 mL Yue alcohol, stirring is lower to add Yue sodium alkoxide（29 mg, 0.54 mmol), stir 10 minutes, revolving removes solvent and dry solid title compound (163 mg).
Molecular formula： C23H14F3N6Na206P molecular weight： 604.34 LC-MS(m/e): 560.7 (M+H)
1H-NMR(400MHz, CDC13) δ:9.28 (s, IH), 8.13 ~ 8.30 (m, 3H), 7.92 (s, IH) 7.81 (m,), IH 7.70 (m, 2H), 6.76 (m, IH), 6.48 (s, 2H), 6.24 (m, IH), the 9- of 5.59 (m, 2H) embodiments 3 (2- aminopyrimidine -5- bases)-l- (3- (trifluoromethyls）Phenyl）Pyrimido【5,4-c l, 5】Naphthyridines-2,4 (li, 3-diketone（Compound B) preparation
By the chloro- 1- of 9- (3- (trifluoro Yue yls) phenyl) pyrimidine [5,4-c] [l, 5] naphthyridines -2,4 (1,3/)-diketone (300 mg, 0.77 mmol), 5- (4,4,5,5- tetramethyls -1,3,2- dioxaborolanes -2- bases) pyrimidine -2- amine（170 mg, 0.77 mmol), potassium carbonate (317 mg, 2.3 mmol) and four（Triphenylphosphine）Palladium (45 mg, 0.04 mmol) is added in 40 mL dioxane and 2 mL water, and the h of back flow reaction 18, is cooled to room temperature under nitrogen protection, concentration, is added 400mL water, is filtrated to get crude product.Crude product is dissolved in 20mL 6M concentrated hydrochloric acids, washed with dichloro Yue alkane（4 x 50mL), aqueous phase is added drop-wise in aqueous sodium carbonate, filtered, washing, crude product washs with methanol, obtain 9- (2- aminopyrimidine -5- bases) -1- (3- (trifluoro Yue yls) phenyl) pyrimidine [5, 4-c] and [l, 5] naphthyridines -2,4 (1,3/)-diketone（160 mg).
Molecular formula： C2iH12F3N702 LC-MS(m/e): 452.1 (M+H)
^-NMR (400 MHz, DMSO-d6) δ:(12.25 br. s., IH), 9.29 (s, IH), 8.38 (d, IH), 8.24 (d, J=9.2, IH), (8.00 s, 2H), 7.70-7.84 (m, 4H), 7.14 (s, 2H) embodiments 4 (9- 2- aminopyrimidine -5- bases) -2,4- dioxo-W3- (trifluoromethyls）Phenyl)-1,2- dihydro-pyrimidins simultaneously " 5,4-cl " l, the preparation of (4-yl) the methyl phosphorodithioate disodium (compound 14) of 51 naphthyridines-3
1) (small (3- (trifluoromethyl) the phenyl) -1,2- dihydro-pyrimidins of 9- (2- aminopyrimidine -5- bases) -2,4- dioxos simultaneously [5,4-c][l,5] naphthyridines -3 (4 /)-yl) and methyl di-t-butyl phosphate preparation
By 9- (2- aminopyrimidine -5- bases) -1- (3- (trifluoro Yue yls) phenyl) pyrimido [5,4-c] [l, 5] naphthyridines
- 2,4 (1,3 /)-diketone（400 mg, 0.89 mmol) and potassium carbonate (491 mg, 3.56 mmol) be dissolved in 30 mL DMSO, be heated to 60 °C, be added dropwise to di-t-butyl chloromethyl phosphate（298 mg, 1.15 mmol), 60 °C are reacted 30 minutes, are cooled to room temperature, are poured into water, and separate out solid filtering, and vacuum drying obtains yellow solid title compound (450 mg crude products）, it is not purified to be directly used in the next step.
2) (9- (2- aminopyrimidine-5- bases)-2,4- dioxos-1-(3- (trifluoromethyl) phenyl)-1,2- dihydro-pyrimidins [5,4-c | [l,5] naphthyridines -3 (4)-base）The preparation of methyl Er Qing Lin acid esters
Will (9- (2- aminopyrimidine -5- bases) -2,4- dioxo -1- (3- (trifluoro Yue yls) phenyl) -1,2- dihydro-pyrimidin simultaneously [5,4-c] [l, 5] naphthyridines -3 (4 /)-base）Methyl di-t-butyl phosphate (450 mg crude products）It is suspended in dichloromethane (10 mL), adds trifluoroacetic acid (10 mL), be stirred at room temperature 3 hours, is concentrated in vacuo, gained crude product reversed phase column chromatography（Acetonitrile：Water=0 ~ 40%) obtain pale yellow oil, adds a small amount of methanol, and gained solid filtering, methanol washing, vacuum drying obtains faint yellow solid shape title compound (10 mg, two step yields: 2%).
3) (small (3- (trifluoromethyl) phenyl) -1, the 2- dihydros of 9- (2- aminopyrimidine -5- bases) -2,4- dioxos are phonetic Pyridine and 5,4-c] [l, 5] naphthyridines -3 (4)-yl) and methyl phosphorodithioate disodium preparation
Will (9- (2- aminopyrimidine -5- bases) -2,4- dioxo -1- (3- (trifluoromethyl) phenyl) -1,2- dihydro-pyrimidin simultaneously [5,4-c] [l, 5] naphthyridines -3 (4)-base）Methyl dihydrogen phosphoric acid ester（10 mg, 0.018 mmol) it is scattered in methanol（10 mL) in, add the methanol solution of Yue sodium alkoxide（Mass fraction 2%, 96 mg, 0.036 mmol), stir 10 minutes, resulting solution is concentrated in vacuo, and obtains faint yellow solid shape title compound（12 mg).
Molecular formula： C22H13F3N7Na206P molecular weight： 605.3 LC-MS(M/e): 562.1
- Ν Μ Κ (400 MHz, MeOD-^) δ 9.48 (s, 1H), 8.55 (s, 1H), 8.39 (d,
J=8.8 Hz, 1H), 8.15 (d, J=9.2Hz, 1H), 8.07 (s, 2H), 7.80-7.82 (m, 1H), 7.72-7.77 (m, 3H), 2- (4- (9- (aminopyridine -3- the bases of 5.93 (d, J=8.8 Hz, 2H) embodiments 5）- 2,4- dioxo -3,4- dihydro-pyrimidin【5,4-c】[l, 5] naphthyridines -1 (2)-yl) phenyl) -2- methyl propionitrile (compound C) preparation
1) 2- methyl -2- (4- propionitrile
By 2- (4- nitrobenzophenones）Acetonitrile (4.86 g, 30 mmol) is dissolved in 50mL dichloromethane, and 3011^ sodium hydroxides are added dropwise thereto（3.6 g, 90 mmol) the aqueous solution, iodomethane is then added dropwise thereto（10.65 g, 75 mmol).After completion of dropping, lucifuge is reacted 16 hours at room temperature.Add 50 mL water and l OO mL dichloromethane, divide liquid, aqueous phase is extracted with 100 mL dichloro Yue alkane, merge organic phase, dried with the sour sodium of anhydrous ^ L, concentration, crude product obtains faint yellow solid 2- methyl -2- (4- nitrobenzophenones) propionitrile (4.5 g) by silica gel column chromatography (EtOAc/PE=0 ~ l/20)
2) 2- (4- aminophenyls) -2- methyl propionitrile
By 2- methyl -2- (4- nitrobenzophenones）Propionitrile (4.5 g, 23.6 mmol) it is added in reaction bulb, sequentially add Pd/C (450 mg) and 50 mL ethyl acetate, system is vacuumized, hydrogen is passed through, is reacted 15 hours at room temperature, gained reaction solution is filtered by diatomite, washed with ethyl acetate, gained filtrate is concentrated to give colorless oil 2- (4- aminophenyls) -2- methyl propionitrile (3.5 g).
3) the chloro- 4- of 6- ((4- (2- cyano group propyl- 2- yls) phenyl) amino) -1,5- pyridine -3- carboxylic acid, ethyl esters
By 4,6- bis- chloro- 1,5- naphthyridines -3- carboxylic acid, ethyl esters (5.9 g, 21.9 mmol) and 2- (4- aminophenyls) -2- methyl propionitrile (3.5 g, 21.9 mmol) it is dissolved in the lOOmL tert-butyl alcohols, potassium carbonate (6.0 g, 43.8 mmol) is added, back flow reaction is heated to 15 hours.Reaction solution is cooled to after room temperature and is concentrated under reduced pressure, residue adds 100 mL water and 100 mL DCM, divide liquid, aqueous phase is extracted with lOO mL DCM, merge organic phase, with anhydrous sodium sulfate drying, concentration, crude product obtains the chloro- 4- of faint yellow solid 6- ((4- (2- cyano group propyl- 2- bases by silica gel column chromatography (EtOAc/PE=0 l/2)）Phenyl）Amino) -1,5- naphthyridines -3- carboxylic acid, ethyl esters (6.9 g).
4) the chloro- 4- of 6- ((4- (2- cyano group propyl- 2- yls) phenyl) amino)-(4- methoxybenzyls) -1,5- naphthyridines -3-
By the chloro- 4- of 6- ((4- (2- cyano group propyl- 2- yls) phenyl) amino) -1,5- naphthyridines -3- carboxylic acid, ethyl esters (6.9 g, 17.5 mmol) it is dissolved in 50mL Yue alcohol and 50mL tetrahydrofurans, the aqueous solution of 50mL lithium hydroxides (2.2 g, 52.4 mmol) is added dropwise at room temperature.After completion of dropping, react 4 hours at room temperature.Reaction solution is concentrated, 200mL water is added, with salt acid for adjusting pH value to 2 ~ 3, gained solid is filtered, vacuum drying obtains yellow solid.Then it is scattered in l OOmL dichloro Yue alkane, is added 0.05mL DMF.Under ice bath, oxalyl chloride (4.2 g are added dropwise thereto, 33.3 mmol), reaction 4 hours is warmed to room temperature after completion of dropping, remove solvent under reduced pressure and dissolved with lOOmL dichloro Yue alkane, the mixture of triethylamine (5.0 g, 49.8 mmol) and 4-Methoxybenzylamine (2.7 g, 19.9 mmol) is instilled under water-bath.After completion of dropping, reaction solution reacts 15 hours at room temperature, add 50 mL water and lOO mL DCM, divide liquid, aqueous phase is extracted with 100 mL DCM, merges organic phase, with anhydrous sodium sulfate drying, concentration, crude product silica gel column chromatography (EtOAc/PE=l/10 ~ 3/2) obtains faint yellow solid (4.0 g).
5) 2- (4- (the chloro- 3- of 9- (4- methoxybenzyls) -2,4- dioxo -3,4- dihydro-pyrimidins [5,4-c] [l, 5] naphthalene -1 (2 /)-yl) phenyl) -2- Yue base propionitrile
By sodium hydride（1.0 g, 25 mmol) it is suspended in 25 mL DMF, slowly instill 6- chloro- 4- ((4- (2- cyano group propyl- 2- yls) phenyl) amino)-Λ Κ 4- Yue oxygen benzyl thereto at room temperature) -1, the DMF solution of 5- naphthyridines -3- formamides (2.43 g, 5 mmol)（25 mL), 60 °C are warming up to after completion of dropping and is reacted 1 hour.By chlorine Yue acetoacetic esters under water-bath（1.36 g, 12.5 mmol) Slow is added dropwise in reaction solution slowly, and 60 °C are warming up to after completion of dropping and is reacted 16 hours.It is cooled to Slow after room temperature to be poured into water slowly, is extracted with ethyl acetate (3<150mL), merge organic phase, use saturated common salt water washing, with anhydrous sodium sulfate drying, concentration, crude product obtains faint yellow solid (1.2 g) by silica gel column chromatography (EtOAc/PE=0 ~ 1/5).
6) 2- (4- (chloro- 2,4- dioxos -3,4- dihydro-pyrimidin [5,4-c] [l, 5] naphthyridines -1 (2 /)-yls of 9-) phenyl) -2-
By 2- (4- (the chloro- 3- of 9- (4- Yue oxygen benzyl) -2,4- dioxos -3,4- dihydro-pyrimidin [5,4-c] [l, 5] naphthyridines -1 (2/)-yl) phenyl) -2- Yue base propionitrile（1.2 g, 2.3 mmol) it is dissolved in acetonitrile (80 mL) and water (20mL), ammonium ceric nitrate (5.1 g, 9.4 mmol) is added portionwise at room temperature.Reaction solution exists Concentrated after reacting 16 hours at room temperature, add 100 mL water, filtering, solid EA and PE (1:1) wash, vacuum drying obtains yellow solid (600 mg：).
7) 2- (4- (9- (6- aminopyridine -3- bases) -2,4- dioxo -3,4- dihydro-pyrimidins [5,4-c] [l, 5] naphthyridines -1 2 /)-yl) phenyl) -2- methyl propionitrile
By 2- (4- (9- chloro- 2,4- dioxo -3,4- dihydro-pyrimidins [5,4-c] [l, 5] naphthyridines -1 (2 base) phenyl) -2- methyl propionitrile (392 mg, 1.0 mmol), 5- (4,4,5,5- tetramethyl -1,3,2- dioxaborolanes -2- bases) pyridine -2- amine（440 mg, 1.0 mmol, content 50%) and four（Triphenylphosphine) palladium (46 mg, 0.04 mmol) is dissolved in 40 mL dioxane, is charged with potassium carbonate (276 mg, 2 mmol) the 1 mL aqueous solution.Reacted 15 ~ 18 hours in 100 °C under nitrogen protection, be then cooled to room temperature, concentrated; filtering; washing, crude product is washed with ethyl acetate, methanol successively, obtains faint yellow solid 2- (4- (9- (6- aminopyridine -3- bases）- 2,4- dioxo -3,4- dihydro-pyrimidin [5,4-c] [l, 5] naphthyridines -1 (2)-yl) phenyl) -2- methyl propionitrile（150 mg).
Molecular formula： C25H19N702' LC-MS(m/e): 450.1(M+H) iH-NMR OOMz, DMSO-ί ί) δ:12.19 (br. s, 1H), 9.24 (s, 1H), 8.30 (d, J=8.8,1H), 8.17 (d, J=8.8,1H), 7.97 (m, 1H), 7.59 (m, 2H), 7.42 (m, 2H), 7.20 (m, 1H), 6.32 ~ 6.40 (m, 3H), (9- (6- aminopyridine -3- bases)-l- (4- (2- dicyanopropane -2- bases) phenyl) -2, the 4- dioxos -1 of 1.73 (s, 6H) embodiments 6,2- dihydro-pyrimidins simultaneously [5,4-c】[l, 5】Naphthyridines -3 (4H)-base）Methyl phosphorodithioate disodium（Compound 31) preparation
1) (9- (6- aminopyridine -3- bases) -1- (4- (2- dicyanopropane -2- bases）Phenyl) -2,4- dioxo -12- dihydro-pyrimidins simultaneously [5,4-c] [l, 5] naphthyridines -3 (4 /)-base）The preparation of methyl di-t-butyl phosphate
By 2- (4- (9- (6- aminopyridine -3- bases) -2,4- dioxos -3,4- dihydro-pyrimidins simultaneously [5,4-c] [l, 5] naphthyridines -1 (2)-yl) phenyl) -2- methyl propionitrile (60 mg, 0.134 mmol) and potassium carbonate（37 mg, 0.268 mmol) it is added in 6mL dimethyl sulfoxides, it is heated to 50.C.When solution is clarified completely, di-t-butyl chloromethyl phosphate (62 mg, 0.24 mmol) is added dropwise in solution, continues to react lh, LC-MS detection reactions are completed.Reaction solution is poured into 10mL water, filtered, obtained solid is dried, and weigh to obtain crude product 50mg, yield： 55.6%.
2) (9- (6- aminopyridine -3- bases) -1- (4- (2- dicyanopropane -2- bases）Phenyl) -2,4- dioxos -1,2- dihydro-pyrimidin simultaneously [5,4-c] [l, 5] naphthyridines -3 (4 /)-base）The preparation of methyl dihydrogen phosphoric acid ester
By (9- (6- aminopyridine -3- bases)-(4- (2- dicyanopropane -2- bases）Phenyl) -2,4- dioxos -1,2- dihydro-pyrimidin simultaneously [5,4-c] [l, 5] naphthyridines -3 (4/ /)-base）Methyl di-t-butyl phosphate (50 mg, 0.075 mmol) is dissolved in the mixed solution of 5mL trifluoroacetic acids and dichloromethane (volume ratio 1:1) lh, is stirred at room temperature.LC-MS detection reactions are completed, and solvent are spin-dried for, the inverted preparation chromatographic isolation of residue obtains product 3mg, yield 7.1%.
3) (9- (6- aminopyridine-3- bases)-1-(4- (2- dicyanopropane-2- bases) phenyl)-2,4- dioxos-- c] [l, 5] naphthyridines-3 (4)-base）The preparation of Yue base disodium phosphates
By (9- (6- aminopyridine -3- bases) -1- (4- (2- dicyanopropane -2- bases）Phenyl) -2,4- dioxos -1,2- dihydro-pyrimidin simultaneously [5,4-c] [l,5] naphthyridines -3 (4)-base）Methyl dihydrogen phosphoric acid ester (3Mg, 0.0054 mmol) it is dissolved in 5mL Yue sodium alkoxide (0.6 mg, 0.011 mmol) Yue alcoholic solutions, 0.5h is stirred, filtering, solution is spin-dried for, and obtains 2 mg products, yield: 61.5%.
Molecular formula： C26H20N7Na2O6P molecular weight： 603.43 LC-MS(M/e): 560.2(M+H+)
!H-NMR (400 MHz, MeOD) δ:9.45 (s, lH), 8.31 (d, 1H, J=8.8Hz), .12 (d, 2H, J=9.2Hz), 7.63 (d, 2H, J=8.4Hz), 7.48 (d, 2U, J=8.8Hz) .36-7.38 (m, IH), 6,53 (d, IH, J=8.8Hz), 5.90 (d, 2H, J=9.6Hz), 2.66 (s, H), 1.71 (s, 6H)

Claims (9)

  1. Claim
    1st, formula（I compound or its pharmaceutically acceptable salt or alloisomerism shown in)
    A and B are separately CR6, R6For hydrogen, halogen atom, cyano group, hydroxyl, carboxyl ,-(C)nNR8aR8b、 -(CH2)nC(0)R9、 -(CH2)nC(0)NR8aR8b、 -(CH2)nOC(0)R9、 -(CH2)nC(0)(CH2)nOR9、 -(CH2)nN(R8a)C(0)R9, or optionally by 1-3 alkyl, the d_ replaced selected from halogen atom, hydroxyl, carboxyl6Alkoxy；
    R1For hydrogen, or optionally by 1-5 R7aSubstituted alkyl, C2-8Alkenyl, C2-8Alkynyl, C3-8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles base, 7-12 member loop coils base, 7-12 member bridged ring bases；
    R2For hydrogen, or optionally by 1-5 R7bSubstituted C1-6Alkyl, C2.8Alkenyl, C2-8Alkynyl, C3-8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles base, 7-12 member loop coils base, 7-12 member bridged ring bases；
    R3For hydrogen, or optionally by the 1-3 d. replaced selected from halogen atom, hydroxyl, carboxyl6Alkyl； R4、 R5It is separately hydrogen, or optionally by the 1-3 d. replaced selected from halogen atom, hydroxyl, carboxyl6Alkyl；
    M is 1 ~ 3；
    E is hydrogen, or for can be with phosphoric acid forming salt inorganic base or the cation of organic base；
    R7a、 R7bIt is separately
    (1) halogen atom, cyano group, hydroxyl, trifluoro Yue bases ,-(C)nNR8aR8b、 -(CH2)nC(0)R9、 -(CH2)nSR9、 -(CH2)nS(0)2R9、 -(CH2)nS(0)2NR8aR8b、 -(CH2)nN(R8a)S(0)2R9、 -(CH2)nC(0)NR8aR8b 、 -(CH2)nOC(0)R9 、 -(CH2)nC(0)(CH2)nOR9 、 -(CH2)nN(R8a)C(0)R9,
    (2) C optionally replaced by the plain atoms of 1-3 Xuan Zi Halogen, hydroxyl, cyano group, trifluoromethyl1-6Alkyl, C2.8Alkenyl, C2-8Alkynyl, C1-6Alkoxy；
    (3) halogen atom, hydroxyl, cyano group, trifluoromethyl, C optionally are selected from by 1-31-6Alkyl, C2-8Alkenyl, C2-8Alkynyl, d.6Alkoxy ,-(CH2)nNR8aR8b、 -(CH2)nC(0)R9、 -(CH2)nSR9、 -(CH2)nS(0)2R9 -(CH2)nS(0)2NR8aR8b、 -(CH2)nN(R8a)S(0)2R9、 -(CH2)nC(0)N 8aR8b 、 -(CH2)nOC(0)R9 、 -(CH2)nC(0)(CH2)nOR9 、 -(CH2)nN(R8a)C(0)R9Substituted C3.8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles bases；
    R8a、 R8bIt is separately hydrogen, or optionally by 1-3 hydroxyl, halogen atom, cyano group, carboxyl ,-(CH2)nNR8aR8Amino-sulfonyl, amino Yue acyl groups, sulfoamido substitution alkyl, C3.8Cycloalkyl, 6-14 members aryl, 5-14 unit's heteroaryls, 3-14 circle heterocycles bases；
    R9For hydrogen, or optionally by 1-3 selected from halogen atom, cyano group, hydroxyl, carboxyl, '-(CH2)nNR8aRsb, sulfonyl, carbamoyl substitution d-6Alkyl, C1-6Alkoxy；N is 0 ~ 4.
    2nd, compound as claimed in claim 1 or its pharmaceutically acceptable salt or stereoisomer：
    A and B are separately CR6, R6For hydrogen, or optionally by the 1-3 alkyl replaced selected from halogen atom, hydroxyl, carboxyl；
    R1For optionally by 1-3 R7aSubstituted 6-10 members aryl, the single heteroaryl of 5-6 members, the thick heteroaryl of 9-10 members, the single heterocyclic radical of 5-6 members, 9-10 member condensed hetero ring bases；
    R2For optionally by 3 R7bSubstituted 6-10 members aryl, the single heteroaryl of 5-6 members, the thick heteroaryl of 9-10 members, the single heterocyclic radical of 5-6 members, 9-10 member condensed hetero ring bases；
    R3For hydrogen；
    R4、 R5It is separately hydrogen, or C1-6Alkyl；
    M is 1-3;
    E is hydrogen, or for can be with phosphoric acid forming salt inorganic base or the metal cation of organic base；
    R7a、 R7bIt is separately
    (1) halogen atom, cyano group, hydroxyl, trifluoromethyl ,-(CH2)nN 8aR8b、-(CH2)nC(0)R9、 -(CH2)nSR9、 -(CH2)nS(0)2R9、 -(CH2)nS(0)2NR8aR8b、 -(CH2)nN(R8a)S(0)2R9、 -(CH2)nC(0)NR8aR8b 、 -(CH2)nOC(0)R9 、 -(CH2)nC(0)(CH2)nOR9 、 -(CH2)nN(R8a)C(0)R9,
    (2) optionally by 1-3 alkyl, the alkoxies replaced selected from halogen atom, hydroxyl, cyano group, trifluoromethyl；
    (3) halogen atom, hydroxyl, cyano group, trifluoromethyl, alkyl, alkoxy ,-(CH optionally are selected from by 1-32)nNR8aR8b、 -(CH2)nC(0)R9、 -(CH2)nSR9、 -(CH2)nS(0)2R9、 -(CH2)nS(0)2NR8aR8b 、 -(CH2)nN(R8a)S(0)2R9 、 -(CH2)nC(0)NR8aR8b 、 -(CH2)nOC(0)R9、 -(CH2)nC(0)(CH2)nOR9、 -(CH2)nN(R8a)C(0)R9Substituted C3.8Cycloalkyl, 5-10 unit's heteroaryls, 5-10 circle heterocycles bases；
    R8a、 R8bIt is separately hydrogen, or (^6 alkyl；
    R9For hydrogen, or C1-6Alkyl； ,
    N is 0 ~ 2.
    3rd, compound as claimed in claim 2 or its pharmaceutically acceptable salt or stereoisomer：
    A and B are separately CR6, R6For hydrogen, or alkyl；
    R R5It is separately hydrogen；
    M is 1;
    E is hydrogen, or sodium ion.
    4th, compound as claimed in claim 3 or its pharmaceutically acceptable salt or stereoisomer：
    R1For optionally by 1-3 R7aSubstituted phenyl, the single heteroaryl of 5-6 members；
    R2For optionally by 1-3 R7bSubstituted phenyl, the single heteroaryl of 5-6 members, the thick heteroaryl of 9-10 members；
    (1) halogen atom, cyano group, hydroxyl, trifluoromethyl ,-NR8aR8b、 -C(0)R9、 -C(0)NR8aR8b、 -OC(0)R9、 -N(R8a)C(0)R9,
    (2) optionally by 1-3 alkyl, the alkoxies replaced selected from halogen atom, hydroxyl, cyano group, trifluoromethyl,
    (3) optionally by the plain atoms of 1-3 Xuan Zi Halogen, hydroxyl, cyano group, trifluoro Yue bases, d-6Alkyl, alkoxy ,-N8aR8b、 -C(0)R9、 -C(0)NR8aR8b、 -OC(0)R9、 -N(R8a)C(0)R9The substituted single heterocyclic radical of 5-6 members；
    R7bFor
    (1) halogen atom, hydroxyl, trifluoromethyl ,-NR8aR8b、 -C(0)R9、 -SR9、 -S(0)2R9、 -C(0)NR8aR8b、 -OC(0) R9、 -N(R8a)C(0)R9 ,
    (2) optionally by 1-3 alkyl, C replaced selected from halogen atom, hydroxyl, cyano group, trifluoro Yue bases1-6Alkoxy；
    (3) halogen atom, hydroxyl, cyano group, trifluoromethyl, C optionally are selected from by 1-31-6Alkyl, alkoxy ,-NR8aR8b、 -C(0)R9、 -C(0)N 8aR8b、 -OC(0)R9、 -N(R8a)C(0)R9 The first single heteroaryls of substituted 5-6, the single heterocyclic radical of 5-6 members；
    R8a、 R8bIt is separately hydrogen, or C^6Alkyl；
    R9For hydrogen, or C1-6Alkyl.
    5th, compound as claimed in claim 4 or its pharmaceutically acceptable salt or stereoisomer：
    R1For optionally by 1-3 R7aSubstituted phenyl, pyridine radicals, pyrimidine radicals；
    R2For optionally by 1-3 1715Substituted phenyl, pyridine radicals, pyrimidine radicals, thienyl, pyrazolyl, indazolyl, Yin tastes base, pyrido pyrrole radicals, pyrrolo- pyrrole1¾^、 （There is this group in power 6, miss herein）, Pyrazolopyridine base, can quinoline base；
    (1) halogen atom, cyano group, hydroxyl, trifluoromethyl ,-NH2,
    (2) optionally by the 1-3 C replaced selected from halogen atom, hydroxyl, cyano group, trifluoro Yue bases1-4Alkyl,
    (3) optionally by 1-3 selected from halogen atom, hydroxyl, trifluoro Yue bases, alkyl, d.4Piperidyl, the piperazinyl of alkoxy substitution；
    (1) halogen atom, hydroxyl, trifluoromethyl ,-NH2、 -C(0)R9、 -SR9、 -S(0)2R9 -NHC(0)R9,
    (2) optionally replaced by the plain atoms of 1-3 Xuan Zi Halogen, hydroxyl, cyano group, trifluoro Yue bases<^_4Alkyl, C alkoxies；
    (3) optionally by 1-3 selected from halogen atom, hydroxyl, trifluoromethyl, C " alkyl, d.4Alkoxy ,-NH2Substituted pyrrole radicals, pyrazolyl, imidazole radicals, piperidyl, piperazinyl, morpholinyl；
    R9For hydrogen, or alkyl.
    6th, compound as claimed in claim 5 or its pharmaceutically acceptable salt or stereoisomer：
    R1For optionally by 1-3 173Substituted phenyl, pyridine radicals；
    R2For optionally by 1-3 R7bSubstituted phenyl, pyridine radicals, pyrimidine radicals, thienyl, pyrazolyl, indazolyl, Yin tastes base, pyrrole57Each simultaneously pyrrole of fixed and pyrrole Lip river base, pyrrolepDetermine base, pyrazolo pyrrole1^J^, full quinoline base；
    R7aFor
    (1) halogen atom, cyano group, hydroxyl, trifluoromethyl ,-NH2, (2) optionally by 1-2 Yue bases, ethyl, the isopropyls replaced selected from hydroxyl, cyano group, trifluoromethyl,
    (3) hydroxyl, cyano group, trifluoromethyl, methyl, the piperidyl of methoxy substitution, piperazinyl optionally are selected from by 1-2；
    R7bFor
    (1) halogen atom, cyano group, hydroxyl, trifluoromethyl ,-NH2、 -SR9、 -S(0)2R9、 -NHC(0)R9,
    (2) optionally by 1-2 methyl, ethyl, n-propyl, isopropyl, methoxyl group, the ethyoxyls replaced selected from hydroxyl, cyano group, trifluoromethyl；
    (3) hydroxyl, cyano group, trifluoromethyl, methyl, ethyl, Yue epoxides, ethyoxyl ,-NH optionally are selected from by 1-22Substituted pyrrole radicals, pyrazolyl, piperazinyl, morpholinyl；
    R9For hydrogen, methyl, or ethyl.
    7th, compound as claimed in claim 1 or its pharmaceutically acceptable salt or alloisomerism
    6fr
    £f£000/M0ZN3/X3d 05
    Please ZNDALOd 9Z0^Sl/M0i OAV  
    / moooHu/Jld
    £5
    C1-fOOO/1-TOlN3/X3d 9Z0tSl/W0Z ΟΛ
    8th, pharmaceutical composition, it contains the compound or its pharmaceutically acceptable salt or stereoisomer and one or more pharmaceutical carriers described in claim 1.
    9th, Yao Wu Group compounds as claimed in claim 8, wherein also comprising one or more antitumor agents and/or immunodepressant, described antitumor agent and immunodepressant are antimetabolite, selected from capecitabine, gemcitabine, pemetrexed disodium；For growth factor receptor inhibitors, selected from pazopanib, Imatinib, Erlotinib, Lapatinib, Gefitinib, ZD6474；For antibody, selected from Trastuzumab, bevacizumab；For mitotic inhibitor, selected from taxol, vinorelbine, docetaxel, Doxorubicin；For antitumor steroids, selected from Letrozole, tamoxifen, fulvestrant, Flutamide, Triptorelin；For alkylating agents, selected from endoxan, mustargen, melphalan, chlorambucil, BCNU；For metal platinum class, selected from carboplatin, cis-platinum, Oxaliplatin；For Topoisomerase inhibitors, selected from topotecan camptothecine, topotecan, Irinotecan；For immunosupress class, fitted selected from everolimus, sirolimus, special cancer；For purine analogue, selected from Ismipur, 6- thioguanines, imuran；For antibioticses, selected from rhzomorph D, daunorubicin, adriamycin, mitoxantrone, bleomycin, plicamycin；For adrenal cortex inhibitor class, selected from aminoglutethimide.
    10, compound described in claim 1, or the purposes of its pharmaceutically acceptable salt or stereoisomer in the medicine for preparing treatment and/or prevention proliferative diseases, the proliferative diseases are cancer or non-cancerous proliferative diseases, the cancer is selected from brain tumor, lung cancer, squamous cell, carcinoma of urinary bladder, stomach cancer, oophoroma, peritoneal cancer, cancer of pancreas, breast cancer, head and neck cancer, cervix cancer, carcinoma of endometrium, the carcinoma of the rectum, liver cancer, kidney, adenocarcinoma of esophagus, esophageal squamous cell carcinoma, solid tumor, prostate cancer, Yue shape gland cancer, female reproductive tract cancer, carcinoma in situ, lymthoma, neurofibromatosis, osteocarcinoma, cutaneum carcinoma, colon cancer, carcinoma of testis, gastrointestinal stromal tumor, mast cell tumor, Huppert's disease, melanoma, glioma or sarcoma；The non-cancerous proliferative diseases are selected from the hyperplasia of prostate of skin or prostate.
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