CN101287725A - DNA-PK inhibitors - Google Patents

Abstract

Compounds of formula: (I) wherein A, B and D are respectively selected from the group consisting of: (i) CH, NH, C; (ii) CH, N, N; and (iii) CH, O, C; the dotted lines represent two double bonds in the appropriate locations; and where Z is selected from S, O, C(=O), CH2 and NH are disclosed for use in inhibiting DNA-PK.

Description

The DNA-PK inhibitor

The present invention relates to as the compound of DNA-PK inhibitor, their purposes and synthetic.

Dna dependent protein kinase (DNA-PK) is and the nuclear serine/threonine protein kitase that is activated after DNA combines.Biological chemistry and genetic data show that this kinases is made up of big catalytic subunit that is called DNA-PKcs and the adjusting component that is called Ku.DNA-PK shown be dna double splitting of chain (DSB) obturator be again the vital component of V (D) J reformer.In addition, recent research has related in multiple other process, has comprised that chromatin Structure is regulated and the DNA-PK component (Smith, G.C.M. and Jackson, S.P., Genes and Dev., 13,916-934 (1999)) of telomere in keeping.

DNA DSBs is considered to the most fatal infringement that cell can run into.In order to struggle with the serious threat that is caused by DNA DSBs, the eukaryotic cell several mechanisms that developed is regulated its reparation.In higher eucaryote, these mechanism mainly are non-homogeneous terminal connect (NHEJ) of DNA, are also referred to as unconventional reorganization.DNA-PK plays a part crucial in this approach.Active the increasing of DNA-PK obtained proof in vitro and in vivo, and with tumour cell to the resistance of IR and difunctional alkylating agent relevant (people such as Muller C., Blood, 92,2213-2219 (1998), people such as Sirzen F., Eur.J.Cancer, 35,111-116 (1999)).Therefore, the resistance mechanism of the active increase of DNA-PK as cell and tumour proposed.Therefore, suppressing DNA-PK with micromolecular inhibitor can prove and think that to wherein cross expressing the tumour of resistance mechanism is effective.

Also had been found that PI3-kinase inhibitor LY294002 in the past

Can be in the function (Izzard, people such as R.A., Cancer Res., 59,2581-2586 (1999)) of vitro inhibition DNA-PK.LY294002 is～1 μ M to the IC50 (making the concentration of 50% enzymic activity forfeiture) of DNA-PK, with identical to the kinase whose IC50 of PI 3-.In addition, show that LY294002 can also faintly make the effect sensitivity (Rosenzweig, people such as K.E., Clin.Cancer Res., 3,1149-1156 (1999)) of cell to IR.

WO 03/024949 has described the numerous compounds categories as the DNA-PK inhibitor, comprises the following 2-amino-chromene of universal architecture-4-ketone:

Wherein:

It is an example.The IC that this compound exhibits goes out ₅₀Be 10-12nM, SER is 1.3 (100nM) (seeing following described to method).

Other example of DNA-PK inhibitor comprises 1 (2-hydroxyl-4-morpholine-4-base-phenyl)-ethyl ketone (Kashishian, people such as A., Mol.Cancer Ther, 2,1257-1264 (2003)):

And SU11752 (Ismail, people such as I.H., Oncogene, 23,873-882 (2004)):

Participate in the repair process of DNA in view of DNA-PK, and micromolecular inhibitor has shown the mammalian cell radiation-sensitive and the chemical-sensitive that can make cultivation, then uses specific DNA-PK inhibition medicine and will can be used as the promoting agent that can strengthen cancer chemotherapeutic and radiocurable usefulness.The DNA-PK inhibitor can also prove the disease that can be used for treating retrovirus-mediated method.For example, verified, the active forfeiture of DNA-PK can seriously suppress the process that retrovirus integrates people such as (, Science, 284,644-7 (1999)) Daniel R.

The present invention has had now found that the DNA-PK that demonstrates similar level or level raising suppresses, has simultaneously as other useful quality, the particularly solvability of active medicine and the additional compounds of cell usefulness raising.

Therefore, a first aspect of the present invention provides formula I compound and isomer, salt, solvate, chemical protected form and prodrug:

Wherein:

A, B and D are selected from respectively:

(i)CH、NH、C；

(ii) CH, N, N; With

(iii)CH、O、C；

Dotted line is illustrated in two two keys on the suitable location;

R ^N1And R ^N2Be independently selected from hydrogen, optional substituted C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl perhaps can form the optional substituted heterocycle with 4 to 8 annular atomses with the nitrogen-atoms that they connect;

If A, B, D are selected from above group (i), (ii), then Z be selected from S, O, C (=O), CH ₂And NH; If A, B, D represent group (iii), then Z be selected from O, C (=O), CH ₂And NH;

R ⁴Be selected from H, OH, NO ₂, NH ₂And Q-Y-X, wherein

Q is-NH-C (=O)-or-O-;

Y is optional substituted C _1-5Alkylene;

X is selected from SR ^S1Or NR ^N3R ^N4, wherein

R ^S1Perhaps R ^N3And R ^N4Be independently selected from hydrogen, optional substituted C _1-7Alkyl, C _5-20Aryl or C _3-20Heterocyclic radical, perhaps R ^N3And R ^N4Can form optional substituted heterocycle with the nitrogen-atoms that they connect with 4 to 8 annular atomses;

If Q is-O-, then X can also be selected from-C (=O)-NR ^N5R ^N6, R wherein ^N5And R ^N6Be independently selected from hydrogen, optional substituted C _1-7Alkyl, C _5-20Aryl or C _3-20Heterocyclic radical, perhaps R ^N5And R ^N6Can form optional substituted heterocycle with the nitrogen-atoms that they connect with 4 to 8 annular atomses, and

If Q is-NH-C (=O)-, then-Y-X can also be selected from C _1-7Alkyl;

Condition is: if A, B, D represent that group (iii) and R ^N1And R ^N2Form morpholino group, then R with their bonded carbon atoms ⁴Cannot be H.

Selection to A, B and D can produce following formula: compound:

A second aspect of the present invention provides the compound that comprises first aspect and the composition of pharmaceutically useful carrier or thinner.

A third aspect of the present invention is provided for the compound of the first aspect of methods of treatment.

The compound that a fourth aspect of the present invention provides first aspect is used for the treatment of by the purposes in the medicine that suppresses the disease that DNA-PK improves in preparation.

The medicine of preferred fourth aspect is compared the activity of the alternative DNA-PK of inhibition with PI 3-kinases and/or ATM.Selectivity is an important problem, because other PI 3-kinases family member's inhibition can cause the disadvantageous side effect relevant with the afunction of these enzymes.

Especially, compound can be used to prepare the medicine that is used for following purpose:

(a) in cancer therapy, be used as adjuvant or be used to strengthen the effectiveness of the treatment of ionizing radiation or chemotherapeutic to tumour cell; Perhaps

(b) disease of treatment retrovirus-mediated method.

Another aspect of the present invention is provided for treating the active compound as described herein of the method for human body or animal body, is preferably the form of pharmaceutical composition.

Another aspect of the present invention is provided at the method that suppresses DNA-PK in external or the body, and this method comprises makes cell contact with the active compound as described herein of significant quantity.

Definition

C _1-7Alkyl: term " C used herein _1-7Alkyl " relate to from having the C of 1 to 7 carbon atom _1-7Remove a resulting monovalence part of hydrogen atom in the hydrocarbon compound, it can be aliphatic or alicyclic or its combination, and can be saturated, part is undersaturated or undersaturated fully.

Saturated straight chain C _1-7The example of alkyl includes but not limited to methyl, ethyl, n-propyl, normal-butyl and n-pentyl (amyl group).

Saturated side chain C _1-7The example of alkyl includes but not limited to sec.-propyl, isobutyl-, sec-butyl, the tertiary butyl and neo-pentyl.

Saturated alicyclic C _1-7Alkyl (is also referred to as " C _3-7Cyclic hydrocarbon radical ") example include but not limited to cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; and the group (group that for example comprises this class group) that replaces, for example methyl cyclopropyl, dimethyl cyclopropyl, methyl cyclobutyl, dimethylcyclobutyl, methylcyclopentyl, dimethylcyclopentyl, methylcyclohexyl, Dimethylcyclohexyl, cyclopropyl methyl and cyclohexyl methyl.

Unsaturated C with one or more carbon-to-carbon double bond _1-7Alkyl (is also referred to as " C _2-7Alkenyl ") example include but not limited to vinyl (CH=CH ₂), 2-propenyl (allyl group ,-CH-CH=CH ₂), pseudoallyl (C (CH ₃)=CH ₂), butenyl, pentenyl and hexenyl.

Unsaturated C with one or more carbon-to-carbon, three key _1-7Alkyl (be also referred to as " _C2-7Alkynyl ") example include but not limited to ethynyl and 2-propynyl (propargyl).

Unsaturated cycloaliphatic (carbocyclic ring) C with one or more carbon-to-carbon double bond _1-7Alkyl (is also referred to as " C _3-7Cycloalkenyl group ") example include but not limited to unsubstituted group, for example cyclopropenyl radical, cyclobutene base, cyclopentenyl and cyclohexenyl, and the group (group that for example comprises this class group) that replaces, for example cyclopropenyl radical methyl and cyclohexenyl methyl.

C _3-20Heterocyclic radical: term " C used herein _3-20Heterocyclic radical " relate to from C _3-20Remove a resulting monovalence part of hydrogen atom in the annular atoms of heterogeneous ring compound, described compound has a ring or two or more ring (for example volution, condensed, bridging), and have 3 to 20 annular atomses, wherein 1 to 10 is ring hetero atom, and wherein at least one described ring is a heterocycle.Preferably, each ring has 3 to 7 annular atomses, and wherein 1 to 4 is ring hetero atom.Ring hetero atom can be preferably selected from O, N, S and P." C _3-20" the representative ring atom, no matter annular atoms is carbon atom or heteroatoms.

C with an azo-cycle atom _3-20The example of heterocyclic radical includes but not limited to from ethylene imine, azetidine, tetramethyleneimine (Pyrrolidine), pyrroline (for example 3-pyrroline, 2,5-pyrrolin), 2H-pyrroles or 3H-pyrroles (different pyrroles), piperidines, dihydropyridine, tetrahydropyridine and azepine

Those of giving birth to.

C with an oxygen annular atoms _3-20The example of heterocyclic radical includes but not limited to from oxyethane, trimethylene oxide, tetrahydrofuran (tetrahydrofuran (THF)), oxa-cyclopentenes (oxole) (dihydrofuran), amylene oxide (oxane) (tetrahydropyrans), dihydropyrane, pyrans (C ₆) and oxa-

Deutero-those.The C that replaces _3-20The example of heterocyclic radical comprises the annular form of sugar, and for example furanose and pyranose for example comprise ribose, lyxose, wood sugar, semi-lactosi, sucrose, fructose and pectinose.

C with a sulphur annular atoms _3-20The example of heterocyclic radical include but not limited to from thiirane (thiirane), Thietane (thietane), thiacyclopentane (thiolane) (tetramethylene sulfide), thia hexanaphthene (thiane) (tetrahydric thiapyran) and thia suberane (thiepane) deutero-those.

C with two oxygen annular atomses _3-20The example of heterocyclic radical include but not limited to from dioxolane, diox and Dioxepane (dioxepane) deutero-those.

C with two azo-cycle atoms _3-20The example of heterocyclic radical includes but not limited to from those of imidazolidine, pyrazolidine (diazole alkane), tetrahydroglyoxaline, pyrazoline (pyrazoline) and piperazine derivatives.

C with an azo-cycle atom and an oxygen annular atoms _3-20The example of heterocyclic radical include but not limited to from Si Qing oxazole, dihydro-oxazole, tetrahydrochysene isoxazole, dihydro-isoxazole, morpholine, Si Qing oxazine, Er Qing oxazine He oxazine deutero-those.

C with an oxygen annular atoms and a sulphur annular atoms _3-20The example of heterocyclic radical includes but not limited to from oxathiolane (oxathiolane) and oxathiane (oxathiane) (thioxane) deutero-those.

C with an azo-cycle atom and a sulphur annular atoms _3-20The example of heterocyclic radical include but not limited to from thiazoline, thiazolidine and parathiazan deutero-those.

Other C _3-20The example Bao of heterocyclic radical draw together but Bu Xian Yu oxadiazine with the Evil thiazine.

Carry in addition one or more oxos (=O) example of the heterocyclic radical of group include but not limited to from the following compounds deutero-those:

C ₅Heterocycle, for example furanone (furanone), pyrone, pyrrolidone, pyrazolone, imidazolidone, thiazolinone (thiazolone) and isothiazolinone (isothiazolone);

C ₆Heterocycle, for example piperidone, dioxopiperidine, piperazine ketone, piperazinedione, pyridazinone and pyrimidone (for example cytosine(Cyt), thymus pyrimidine, uridylic) and barbituric acid;

Condensed heterocycle, for example oxindole, hypoxanthine (for example guanine), benzoxazolinone, benzopyrone (for example tonka bean camphor);

Cyclic acid anhydride (in the ring-C (=O)-O-C (=O)-), include but not limited to maleic anhydride, succinyl oxide and Pyroglutaric acid;

Cyclic carbonate (in the ring-O-C (=O)-O-), for example ethylene carbonate and carbonic acid 1, the inferior propyl ester of 2-;

Imide (in the ring-C (=O)-NR-C (=O)-), include but not limited to succinimide, maleimide, phthalimide and glutarimide;

Lactone (cyclic ester, in the ring-O-C (=O)-), include but not limited to beta-propiolactone, gamma-butyrolactone, δ-Wu Neizhi (2-piperidone) and 6-caprolactone;

Lactan (cyclic amide, in the ring-NR-C (=O)-), include but not limited to azetidinone, butyrolactam (2-Pyrrolidone), δ-Valerolactim and ε-Ji Neixianan;

Cyclic carbamate (in the ring-O-C (=O)-NR-), 2-oxazolidone for example;

The ring-type urea (in the ring-NR-C (=O)-NR-), for example 2-imidazolidone and pyrimidine-2,4-diketone (for example thymus pyrimidine, uridylic).

C _5-20Aryl: term " C used herein _5-20Aryl " relate to from C _5-20Remove a resulting monovalence part of hydrogen atom in the aromatic ring atom of aromatic substance, described compound has a ring or two or more ring (for example condensed), and has 5 to 20 annular atomses, and wherein at least one described ring is an aromatic ring.Preferably, each ring has 5 to 7 annular atomses.

Annular atoms can all be a carbon atom, i.e. " carbon aryl ", this group " C that can suit to be called as in this case _5-20Carbon aryl ".

The C that does not have ring hetero atom _5-20Aryl (is C _5-20The carbon aryl) example includes but not limited to from benzene (being phenyl) (C ₆), naphthalene (C ₁₀), anthracene (C ₁₄), luxuriant and rich with fragrance (C ₁₄), tetracene (C ₁₈) and pyrene (C ₁₆) deutero-those.

Comprise fused rings, one of them is not that the example of the aryl of aromatic ring includes but not limited to from indenes and fluorenes deutero-group.

Select as an alternative, annular atoms can comprise one or more heteroatomss, includes but not limited to oxygen, nitrogen and sulphur, as " heteroaryl ".In this case, this group " C that can suit to be called as _5-20Heteroaryl ", " C wherein _5-20" representative ring atom, no matter carbon atom or heteroatoms.Preferably, each ring has 5 to 7 annular atomses, and wherein 0 to 4 is ring hetero atom.

C _5-20The example of heteroaryl includes but not limited to derived from furans (oxole), thiophene, pyrroles, imidazoles (1, the 3-diazole), pyrazoles (1,2-diazole), triazole, oxazole, isoxazole, thiazole, isothiazole, oxadiazole with oxatriazole deutero-C ₅Heteroaryl; With Cong Yi oxazine, pyridine (azine), pyridazine (1,2-diazine), pyrimidine (1,3-diazines; For example cytosine(Cyt), thymus pyrimidine, uridylic), pyrazine (1,4-diazines), triazine, tetrazolium be with oxadiazole (furazan) deutero-C ₆Heteroaryl.

The C that comprises condensed ring _5-20(some of them are C to heterocyclic group _5-20Heteroaryl) example includes but not limited to the C derived from cumarone, isobenzofuran, indoles, isoindole, purine (for example VITAMIN B4, guanine), thionaphthene, benzoglyoxaline ₉Heterocyclic group; C derived from quinoline, isoquinoline 99.9, benzodiazine, pyrrolopyridine, quinoxaline ₁₀Heterocyclic group; C derived from carbazole, dibenzothiophene, diphenylene-oxide ₁₃Heterocyclic group; C derived from acridine, xanthene, phenoxathiin (phenoxathiin), azophenlyene, phenoxazine, thiodiphenylamine ₁₄Heterocyclic group.

Above-mentioned C _1-7Alkyl, C _3-20Heterocyclic radical and C _5-20No matter aryl still is another substituent part separately, and itself can randomly be replaced by one or more groups, and substituting group is selected from their itself and following other substituting group.

Halogeno-group :-F ,-Cl ,-Br and-I.

Hydroxyl :-OH.

Ether :-OR, wherein R is ether substituting group, for example C _1-7Alkyl (is also referred to as C _1-7-oxyl is discussed as follows), C _3-20Heterocyclic radical (is also referred to as C _3-20Heterocyclic oxy group) or C _5-20Aryl (is also referred to as C _5-20Aryloxy), preferred C _1-7Alkyl.

C _1-7-oxyl :-OR, wherein R is C _1-7Alkyl.C _1-7The example of-oxyl includes but not limited to-OCH ₃(methoxyl group) ,-OCH ₂CH ₃(oxyethyl group) and-OC (CH ₃) ₃(tert.-butoxy).

Oxo (ketone group ,-ketone) :=O.Have oxo group (=O) include but not limited to carbocyclic ring, for example cyclopentanone and pimelinketone as substituent ring compound and/or examples of groups; Heterocycle, for example pyrone, pyrrolidone, pyrazolone, pyrazolone, piperidone, dioxopiperidine, piperazinedione and imidazolidone; Cyclic acid anhydride includes but not limited to maleic anhydride and succinyl oxide; Cyclic carbonate, for example Texacar PC; Imide includes but not limited to succinimide and maleimide; Lactone (cyclic ester, in the ring-O-C (=O)-), include but not limited to beta-propiolactone, gamma-butyrolactone, δ-Wu Neizhi and 6-caprolactone; And lactan (cyclic amide, in the ring-NR-C (=O)-), include but not limited to azetidinone, butyrolactam (2-Pyrrolidone), δ-Valerolactim and ε-Ji Neixianan.

Imino-(imines) :=NR, wherein R is the imino-substituting group, for example hydrogen, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred hydrogen or C _1-7Alkyl.The example of ester group includes but not limited to=NH ,=NMe ,=NEt and=NPh.

Formyl radical (aldehyde, formaldehyde) :-C (=O) H.

(=O) R, wherein R is an acyl substituent to acyl group (ketone group) :-C, for example C _1-7Alkyl (is also referred to as C _1-7Alkyl acyl group or C _1-7The hydrocarbon acyl group), C _3-20Heterocyclic radical (is also referred to as C _3-20The heterocyclic radical acyl group) or C _5-20Aryl (is also referred to as C _5-20Aryl-acyl), preferred C _1-7Alkyl.The example of acyl group includes but not limited to-C (=O) CH ₃(ethanoyl) ,-C (=O) CH ₂CH ₃(propionyl) ,-C (=O) C (CH ₃) ₃(butyryl radicals) and-C (=O) Ph (benzoyl, benzophenone).

Carboxyl (carboxylic acid group) :-COOH.

(=O) OR, wherein R is ester substituting group, for example C to ester (ester of carboxylicesters, carboxylic acid, oxygen carbonyl) :-C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of ester group includes but not limited to-C (=O) OCH ₃,-C (=O) OCH ₂CH ₃,-C (=O) OC (CH ₃) ₃With-C (=O) OPh.

(=O) R, wherein R is acyloxy substituting group, for example C to acyloxy (anti-ester) :-OC _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of acyloxy includes but not limited to-OC (=O) CH ₃(acetoxyl group) ,-OC (=O) CH ₂CH ₃,-OC (=O) C (CH ₃) ₃,-OC (=O) Ph and-OC (=O) CH ₂Ph.

Amido (formamyl, carbamyl, aminocarboxyl, acid amides) :-C (=O) NR ¹R ², R wherein ¹And R ²Be amino substituting group independently, such as about amino definition.The example of amido includes but not limited to-C (=O) NH ₂,-C (=O) NHCH ₃,-C (=O) N (CH ₃) ₂,-C (=O) NHCH ₂CH ₃With-C (=O) N (CH ₂CH ₃) ₂, and such amido, wherein R ¹And R ²Constitute heterocycle structure with their accompanying nitrogen-atoms, for example piperidino-(1-position only) carbonyl, morpholino base carbonyl, parathiazan are for basic carbonyl and Piperazino carbonyl.

Acyl group amido (acyl amino) :-NR ¹C (=O) R ², R wherein ¹Be amide substituents, for example hydrogen, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred hydrogen or C _1-7Alkyl, R ²Be acyl substituent, C for example _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred hydrogen or C _1-7Alkyl.The example of acyl group amide group includes but not limited to-NHC (=O) CH ₃,-NHC (=O) CH ₂CH ₃With-NHC (=O) Ph.R ¹And R ²Can constitute ring texture together, for example the inferior hydrogen base of succinyl-, maleimide amino and the adjacent diformazan acylimino of benzene:

The amino phthaloyl imino of succinimido maleimide

Acyl group urea groups :-N (R ¹) C (O) NR ²C (O) R ³, R wherein ¹And R ²Be the urea groups substituting group independently, for example hydrogen, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred hydrogen or C _1-7Alkyl.R ³Be as about the defined acyl group of acyl group.The example of acyl group urea groups includes but not limited to-NHCONHC (O) H ,-NHCONMeC (O) H ,-NHCONEtC (O) H ,-NHCONMeC (O) Me ,-NHCONEtC (O) Et ,-NMeCONHC (O) Et ,-NMeCONHC (O) Me ,-NMeCONHC (O) Et ,-NMeCONMeC (O) Me ,-NMeCONEtC (O) Et and-NMeCONHC (O) Ph.

Carbamate groups :-NR ¹-C (O)-OR ², R wherein ¹Be as about defined amino substituting group of amino and R ²Be as to the defined ester group of ester group.The example of carbamate groups includes but not limited to-NH-C (O)-O-Me ,-NMe-C (O)-O-Me ,-NH-C (O)-O-Et ,-NMe-C (O)-O-tertiary butyl and-NH-C (O)-O-Ph.

Thio acylamino (thiocarbamyl) :-C (=S) NR ¹R ², R wherein ¹And R ²Be amino substituting group independently, such as about amino definition.The example of amido includes but not limited to-C (=S) NH ₂,-C (=S) NHCH ₃,-C (=S) N (CH ₃) ₂With-C (=S) NHCH ₂CH ₃

Tetrazyl: have five yuan of aromatic rings of four nitrogen-atoms and a carbon atom,

Amino :-NR ¹R ², R wherein ¹And R ²Be amino substituting group independently, for example hydrogen, C _1-7Alkyl (is also referred to as C _1-7Amino or the two-C of alkyl _1-7Alkyl amino), C _3-20Heterocyclic radical or C _5-20Aryl, preferred H or C _1-7Alkyl, perhaps under " ring-type " amino situation, R ¹And R ²Constitute heterocycle with the nitrogen-atoms that they connected with 4 to 8 annular atomses.Amino example includes but not limited to-NH ₂,-NHCH ₃,-NHC (CH ₃) ₂,-N (CH ₃) ₂,-N (CH ₂CH ₃) ₂With-NHPh.The example of cyclic amino includes but not limited to that ethylene imine subbase, azetidine subbase, pyrrolidino, piperidino-(1-position only), Piperazino, morpholino base and parathiazan are for base.

Imino-:=NR, wherein R is the imino-substituting group, for example hydrogen, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred H or C _1-7Alkyl.

Amidine :-C (=NR) NR ₂, wherein each R is the amidine substituting group, for example hydrogen, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred H or C _1-7Alkyl.The example of amidino groups is-C (=NH) NH ₂

Carbazoyl (diazanyl carbonyl) :-C (O)-NN-R ¹, R wherein ¹Be as about the defined amino substituting group of amino.The example of azino (azino) includes but not limited to-C (O)-NN-H ,-C (O)-NN-Me ,-C (O)-NN-Et ,-C (O)-NN-Ph and-C (O)-NN-CH ₂-Ph.

Nitro :-NO ₂

Nitroso-group :-NO.

Azido-:-N ₃

Cyano group (itrile group, formonitrile HCN) :-CN.

Isocyano-:-NC.

Cyanato-:-OCN.

Different cyanato-:-NCO.

Thiocyanogen (thiocyano) :-SCN.

Isothiocyano (isocyanide sulfenyl) :-NCS.

Sulfydryl (thiol, thiohydroxy) :-SH.

Thioether (sulfide) :-SR, wherein R is thioether substituting group, for example C _1-7Alkyl (is also referred to as C _1-7Sulfenyl), C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.C _1-7The example of sulfenyl includes but not limited to-SCH ₃With-SCH ₂CH ₃

Disulfide group :-SS-R, wherein R is disulfide group substituting group, for example C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7(this paper also is called C to alkyl _1-7The alkyl disulfide group).C _1-7The example of alkyl disulfide group includes but not limited to-SSCH ₃With-SSCH ₂CH ₃

Sulfo group (alkylsulfonyl) :-S (=O) ₂R, wherein R is sulfo group substituting group, for example C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfo group includes but not limited to-S (=O) ₂CH ₃(methylsulfonyl) ,-S (=O) ₂CF ₃(trifyl) ,-S (=O) ₂CH ₂CH ₃,-S (=O) ₂C ₄F ₉(nine fluorine fourth alkylsulfonyls) ,-S (=O) ₂CH ₂CF ₃(trifluoro ethylsulfonyl) ,-S (=O) ₂Ph (benzenesulfonyl), 4-Methyl benzenesulfonyl base (tosyl group), 4-bromobenzenesulfonyl (bromobenzenesulfonyl) and 4-nitrophenyl (oil of mirbane alkylsulfonyl).

(=O) R, wherein R is sulfino substituting group, for example C to sulfino (sulfinyl, sulfoxide group) :-S _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfino includes but not limited to-S (=O) CH ₃With-S (=O) CH ₂CH ₃

Sulfonyloxy :-OS (=O) ₂R, wherein R is sulfonyloxy substituting group, for example C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfonyloxy includes but not limited to-OS (=O) ₂CH ₃With-OS (=O) ₂CH ₂CH ₃

(=O) R, wherein R is thionyl oxy substituents, for example C to sulfurous acyloxy :-OS _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfurous acyloxy includes but not limited to-OS (=O) CH ₃With-OS (=O) CH ₂CH ₃

Sulfoamino-(Sulfamino) :-NR ¹S (=O) ₂OH, wherein R ¹Be as closing amino defined amino substituting group.The example of sulfoamino-includes but not limited to-NHS (=O) ₂OH and-N (CH ₃) S (=O) ₂OH.

Inferior sulfoamino-:-NR ¹S (=O) R, wherein R ¹Be as about the defined amino substituting group of amino, and R is inferior sulfoamino-substituting group, for example C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of inferior sulfoamino-includes but not limited to-NHS (=O) CH ₃With-N (CH ₃) S (=O) C ₆H ₅

Sulfamyl :-S (=O) NR ¹R ², R wherein ¹And R ²Be as about the defined amino substituting group of amino independently.The example of sulfamyl includes but not limited to-S (=O) NH ₂,-S (=O) NH (CH ₃) ,-S (=O) N (CH ₃) ₂,-S (=O) NH (CH ₂CH ₃) ,-S (=O) N (CH ₂CH ₃) ₂With-S (=O) NHPh.

Sulfonamido :-NR ¹S (=O) ₂R, wherein R ¹Be as about the defined amino substituting group of amino, and R is sulfonamido substituting group, for example C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfonamido includes but not limited to-NHS (=O) ₂CH ₃With-N (CH ₃) S (=O) ₂C ₆H ₅The sulfonamido of special class be derived from sultam those-in these groups, R ¹Be C one of among the R _5-20Aryl, preferred phenyl, and R ¹With another is and C among the R _5-20The bidentate group that aryl connects is for example derived from C _1-7The bidentate group of alkyl.These examples of groups include but not limited to:

2,3-dihydro-benzo [d] isothiazole-1,1-titanium dioxide-2-base

1,3-dihydro-benzo [c] isothiazole-2,2-titanium dioxide-1-base

3,4-dihydro-2H-benzo [e] [1,2] thiazine-1,1-titanium dioxide-2-base

Amino phosphorous acid ester group (Phosphoramidite) :-OP (OR ¹)-NR ² ₂, R wherein ¹And R ²Be amino phosphorous acid ester group substituting group, for example-H, (optional substituted) C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred-H, C _1-7Alkyl or C _5-20Aryl.The example of amino phosphorous acid ester group includes but not limited to-OP (OCH ₂CH ₃)-N (CH ₃) ₂,-OP (OCH ₂CH ₃)-N (i-Pr) ₂, and-OP (OCH ₂CH ₂CN)-N (i-Pr) ₂

Phosphoramidic acid ester group :-OP (=O) (OR ¹)-NR ² ₂, R wherein ¹And R ²Be phosphoramidic acid ester group substituting group, for example-H, (optional substituted) C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred-H, C _1-7Alkyl or C _5-20Aryl.The example of phosphoramidic acid ester group includes but not limited to-OP (=O) (OCH ₂CH ₃)-N (CH ₃) ₂,-OP (=O) (OCH ₂CH ₃)-N (i-Pr) ₂, and-OP (=O) (OCH ₂CH ₂CN)-N (i-Pr) ₂

Under many circumstances, substituting group itself can be substituted.For example, C _1-7-oxyl for example can be by C _1-7The alkyl replacement (is also referred to as C _1-7Alkyl-C _1-7-oxyl), cyclohexyl methoxyl group for example is by C _3-20The heterocyclic radical replacement (is also referred to as C _5-20Aryl-C _1-7-oxyl), the adjacent diformazan acylimino of benzene oxyethyl group for example is perhaps by C _5-20The aryl replacement (is also referred to as C _5-20Aryl-C _1-7-oxyl), benzyloxy for example.

C _1-5Alkylene: term " C as used herein _1-5Alkylene " relate to by same carbon atom and remove two hydrogen atoms or from two different carbon atoms of described aliphatics straight chain hydrocarbon compound, respectively remove two toothed portions that a hydrogen atom obtains from aliphatics straight chain hydrocarbon compound (except as otherwise noted) with 1 to 5 carbon atom, its can be saturated, part is unsaturated or undersaturated fully.Therefore, term " alkylene " comprises subclass alkenylene discussed below, alkynylene etc.

Saturated C _1-5The example of alkylene includes but not limited to-(CH ₂) _n-, wherein n is 1 to 5 integer, for example-and CH ₂-(methylene radical) ,-CH ₂CH ₂-(ethylidene) ,-CH ₂CH ₂CH ₂-(propylidene) and-CH ₂CH ₂CH ₂CH ₂-(butylidene).

The undersaturated C of part _1-5The example of alkylene includes but not limited to-CH=CH-(vinylidene) ,-CH=CH-CH ₂-,-CH ₂-CH=CH ₂-,-CH=CH-CH ₂-CH ₂-,-CH=CH-CH ₂-CH ₂-CH ₂-,-CH=CH-CH=CH-and-CH=CH-CH=CH-CH ₂-.

Above listed substituting group group can be the substituting group on the alkylene.

Comprise other form

Above comprise these substituent ions, salt, solvate and the protected form known.For example, (appellation COOH) also comprises negatively charged ion (carboxylate radical) form (COO-), its salt or solvate and conventional protected form to carboxylic acid.Similarly, the appellation to amino comprises protonated form (N ⁺HR ¹R ²), amino salt or solvate, for example hydrochloride, and amino conventional protected form.Similarly, the appellation to hydroxyl also comprises anionic form (O ^-), the conventional protected form of its salt or solvate and hydroxyl.

Isomer, salt, solvate, protected form and prodrug

Some compound can exist one or more specific how much, optically-active, mapping, non-mapping, epimerization, stereoisomerism, tautomerism, conformation or end group heterogeneous, include but not limited to cis-with trans-type; E-and Z-type; C-, t-and r-type; In-(endo-) with outer (exo)-type; R-, S-and meso-type; D-and L-type; D-and l-type; (+) and (-) type; Ketone group-, enol-with enolate-type; Syn-and anti-type; Synclinal and anticlinal type; α-with β-type, axially with calm type, ship-, chair-, twist-, envelope-with half chair-type; And combination, below be referred to as " isomer " (or " heterogeneous ").

Attention: about the discussion of tautomerism type, especially will be from term used herein " isomer " excluded be structure (or structure) isomer (be the connection between the atom but not only be the differentiated isomer in atoms in space position) except hereinafter.For example, to methoxyl group-OCH ₃Appellation should not be interpreted as relating to its constitutional isomer, i.e. methylol-CH ₂OH.Similarly, the appellation of neighbour-chloro-phenyl-is not interpreted as relating to its constitutional isomer, promptly between-chloro-phenyl-.But, the appellation to a class formation also can comprise Structural Isomerism type (for example, the C that belongs to such _1-7Alkyl comprises n-propyl and sec.-propyl; Butyl just comprising-, different-, secondary-with tert-butyl; P-methoxy-phenyl comprise the neighbour-,-with right-p-methoxy-phenyl).

Above-mentioned eliminating does not relate to tautomerism type, for example ketone group-, enol-with enolate-type, for example following tautomerism is right: ketone group/enol (illustration is as follows), imines/enamine, acid amides/imino-alcohol, amidine/amidine, nitroso-group/oxime, sulfo-ketone/alkene mercaptan, N-nitroso-group/hydroxyl azo and nitro/acid-nitro.

The ketoenoles enolate

Attention: in term " isomer ", will comprise having the compound that one or more isotropic substances replace especially.For example, H can be any isotropic substance form, comprises ¹H, ²H (D) and ³H (T); C can be any isotropic substance form, comprises ¹²C, ¹³C and ¹⁴C; O can be any isotropic substance form, comprises ¹⁶O and ¹⁸O; Deng.

Unless otherwise specified, the appellation of specific compound is comprised all these class heterogeneous, comprise (wholly or in part) its racemize and other mixtures.Preparation method of this class heterogeneous (for example asymmetric synthesis) and separation method (for example fractional crystallization and chromatogram means) are well known in the art or adjust method teaching herein or currently known methods acquisition according to known way easily.

Unless otherwise specified, the appellation of specific compound is also comprised its ion, salt, solvate and protected form, for example as discussed below.

May suit or need to prepare, purifying and/or handle the salt of the correspondence of active compound, for example pharmaceutically useful salt.The example of pharmacologically acceptable salt is people such as Berge, J.Pharm.Sci., and 66,1-19 has discussion in (1977).

For example, if compound be anionic or have can be anionic functional group (for example ,-COOH can be-COO ^-), can generate salt with the positively charged ion that is fit to so.The example of the inorganic cation that is fit to includes but not limited to alkalimetal ion such as Na ⁺And K ⁺, alkaline earth metal cation such as Ca ²⁺And Mg ²⁺And other positively charged ions such as Al ³⁺The organic cations example that is fit to includes but not limited to that ammonium ion (is NH ₄ ⁺) and the ammonium ion that replaces (NH for example ₃R ⁺, NH ₂R ₂ ⁺, NHR ₃ ⁺, NR ₄ ⁺).The example of the ammonium ion of the replacement that some are fit to be from the following compounds deutero-those: ethamine, diethylamine, dicyclohexyl amine, triethylamine, butylamine, quadrol, thanomin, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine and Trometamol, and amino acid, for example Methionin and arginine.The example of common quaternary ammonium ion is N (CH ₃) ₄ ⁺

If compound be cationic or have can be cationic functional group (for example ,-NH ₂Can be-NH ₃ ⁺), can generate salt with the negatively charged ion that is fit to so.The example of the inorganic anion that is fit to include but not limited to from following mineral acid deutero-those: hydrochloric acid, Hydrogen bromide, hydroiodic acid HI, sulfuric acid, sulfurous acid, nitric acid, nitrous acid, phosphoric acid and phosphorous acid.The example of the organic anion that is fit to include but not limited to from following organic acid deutero-those: acetate, propionic acid, succsinic acid, oxyacetic acid, stearic acid, palmitinic acid, lactic acid, oxysuccinic acid, pounce on acid, tartrate, citric acid, glyconic acid, xitix, toxilic acid, hydroxymaleic acid, phenylacetic acid, L-glutamic acid, aspartic acid, phenylformic acid, styracin, pyruvic acid, Whitfield's ointment, sulfanilic acid, the 2-acetoxy-benzoic acid, fumaric acid, phenylbenzimidazole sulfonic acid, toluenesulphonic acids, methylsulfonic acid, ethyl sulfonic acid, ethionic acid, oxalic acid, pantothenic acid, isethionic acid, valeric acid, lactobionic acid and glyconic acid.The anionic example of polymerization that is fit to include but not limited to from following polymeric acid deutero-those: tannic acid, carboxymethyl cellulose.

May suit or need to prepare, purifying and/or handle the solvate of the correspondence of active compound.Term " solvate " is used to represent the title complex of solute (for example salt of active compound, active compound) and solvent in this article on conventional meaning.If solvent is a water, suitable hydrate, for example monohydrate, dihydrate, trihydrate etc. of being called as of solvate so.

May suit or need to prepare, purifying and/or handle the chemoproection form of active compound.Term used herein " chemoproection form " relates to such compound; wherein one or more reactive functional groups are protected to avoid unwanted chemical reaction; that is to say, be protected or blocking group (be also referred to as masked or shelter group or be closed or blocking groups) form.By protective reaction functional group, can involve the reaction of other unprotected reactive functional groups, and not influence protected group; Usually in step subsequently, can remove blocking group, and the rest part of substantial effect molecule not.For example referring to Protective Groups in Organic Synthesis (T.Green and P.Wuts, Wiley, 1999).

For example, hydroxyl can protectedly be ether (OR) or ester (OC (=O) R), for example tertbutyl ether; Benzyl, diphenyl-methyl (diphenyl methyl) or trityl (trityl group) ether; Trimethylsilyl or t-butyldimethylsilyl ether; Or ethanoyl ester (OC (=O) CH ₃,-OAc).

For example, the aldehydes or ketones group can be distinguished protected for acetal or ketone acetal, wherein carbonyl (＞C=O) be converted into diether (＞C (OR) by reaction with for example primary alconol ₂).In the presence of acid, use greatly excessive water, by the hydrolytic action aldehydes or ketones group of regenerating easily.

For example, amine groups can protectedly be acid amides or urea for example, for example methyl nitrosourea (NHCO-CH ₃); Benzyloxy acid amides (NHCO-OCH ₂C ₆H ₅,-NH-Cbz); Tert.-butoxy acid amides (NHCO-OC (CH ₃) ₃,-NH-Boc); 2-biphenyl-2-propoxy-acid amides (NHCO-OC (CH ₃) ₂C ₆H ₄C ₆H ₅,-NH-Bpoc); 9-fluorenyl methoxy acid amides (NH-Fmoc); 6-nitro black false hellebore oxygen base acid amides (NH-Nvoc); 2-trimethylsilyl oxyethyl group acid amides (NH-Teoc); 2,2,2-three chloroethoxy acid amides (NH-Troc); The allyloxy acid amides (NH-Alloc); (2-benzenesulfonyl) the oxyethyl group acid amides (NH-Psec); Perhaps under situation about being fit to, for the N-oxide compound (＞NO).

For example, hydroxy-acid group can protectedly be ester, for example C _1-7Hydrocarbyl carbonate (for example methyl ester, tertiary butyl ester); C _1-7Halo hydrocarbyl carbonate (C for example _1-7Three brine alkyl esters); Three C _1-7Alkyl silyl-C _1-7Hydrocarbyl carbonate; Or C _5-20Aryl-C _1-7Hydrocarbyl carbonate (for example benzyl ester, nitrobenzyl ester); Or acid amides, for example methyl nitrosourea.

For example, the thiol group can protectedly be thioether (SR), a benzyl thioether for example; Acetylamino methyl ether (S-CH ₂NHC (=O) CH ₃).

May suit or need to prepare, purifying and/or handle the prodrug form of active compound.Term used herein " prodrug " relates to a kind of like this compound, and its by metabolism the time (for example in body) produces required active compound.Usually, prodrug is a non-activity, and perhaps activity is weaker than active compound, but favourable processing, administration or metabolisming property can be provided.

For example, some prodrug is the ester (for example acceptable on the physiology, metabolism on unsettled ester) of active compound.Between metabilic stage, ester group (C (=O) OR) is cleaved, produces active medicine.This class ester can generate like this: for example by any esterification of hydroxy-acid group (C (=O) OH) in the parent compound, protect any other reactive groups that exist in the parent compound in advance with asing one sees fit, protect succeeded by going if desired.The example of unsettled ester comprises that R wherein is those of following groups: C in this class metabolism _1-7Alkyl (for example-Me ,-Et); C _1-7Hydrocarbyl amino (for example amino-ethyl, 2-(N, N-diethylin) ethyl, 2-(4-morpholino) ethyl); And acyloxy-C _1-7Alkyl (acyloxy methyl for example, acyloxy ethyl, for example oxy acid methyl neopentyl, acetoxy-methyl, 1-acetoxyl group ethyl, 1-(1-methoxyl group-1-methyl) ethyl-ketonic oxygen base ethyl, 1-(benzoyloxy) ethyl, isopropoxy-ketonic oxygen ylmethyl, 1-isopropoxy-ketonic oxygen base ethyl, cyclohexyl-ketonic oxygen ylmethyl, 1-cyclohexyl-ketonic oxygen base ethyl, cyclohexyloxy-ketonic oxygen ylmethyl, 1-cyclohexyloxy-ketonic oxygen base ethyl, (4-tetrahydro-pyran oxy) ketonic oxygen ylmethyl, 1-(4-tetrahydro-pyran oxy) ketonic oxygen base ethyl, (4-THP trtrahydropyranyl) ketonic oxygen ylmethyl and 1-(4-THP trtrahydropyranyl) ketonic oxygen base ethyl).

In addition, some prodrugs are produced active compound or produce the compound that produces active compound through further chemical reaction by enzyme activation.For example, prodrug can be sugar derivatives or other glucosides conjugate, perhaps can be amino acid ester derivative.

Selectivity suppresses

The degree that ' selectivity inhibition ' expression suppresses a kind of enzyme is bigger than the degree that suppresses one or more other enzymes.This selectivity can be by relatively suppressing a kind of 50% active needed compound concentrations (IC of enzyme ₅₀) with the 50% active needed identical compound concentrations (IC that suppresses other enzyme ₅₀) measure (seeing below).The result is expressed as ratio.If ratio is greater than 1, then test-compound demonstrates certain selectivity in its restraining effect.

Compound of the present invention is compared selectivity that PI 3-kinases preferably demonstrates greater than 3,10,20 or 50 for DNA-PK.

Compound of the present invention is compared selectivity that ATM preferably demonstrates greater than 5,10,50 or 100 for DNA-PK.

Method described in the preferred use WO 03/024949 is measured the IC that is used for Evaluation and Selection ₅₀Value, this patent is introduced into this paper as a reference.

Further preferred

R ⁴

Preferred R ⁴Be Q-Y-X.

When Q be-NH-C (=O)-time, X is NR preferably ^N3R ^N4Preferred in addition Y is optional substituted C _1-3Alkylene, more preferably optional substituted C _1-2Alkylene, and C most preferably _1-2Alkylene.

When Q be-O-and X are NR ^N3R ^N4The time, then Y preferably chooses substituted C wantonly _1-3Alkylene, more preferably optional substituted C _1-2Alkylene, and C most preferably _1-2Alkylene.

In some embodiments, R ^N3And R ^N4Preferably be independently selected from H and optional substituted C _1-7Alkyl, more preferably H and optional substituted C _1-4Alkyl, and most preferably H and optional substituted C _1-3Alkyl (for example methyl, ethyl, sec.-propyl).Preferred optional substituting group include but not limited to hydroxyl, methoxyl group ,-NH ₂, optional substituted C ₆-aryl and optional substituted C _5-6Heterocyclic radical.

In other embodiments, R ^N3And R ^N4The nitrogen-atoms that connects with them forms the optional substituted nitrogen heterocyclic ring with 4 to 8 annular atomses.Preferred heterocycle has 5 to 7 annular atomses.The example of preferred group comprises morpholino base, piperidyl, piperazinyl, high piperazinyl and Pyrrolidine and base (tetrahydropyrrolo), preferred especially piperazinyl.These groups can be substituted, and particularly preferred group is optional substituted piperazinyl, and wherein substituting group is preferably on the 4-nitrogen-atoms.Preferred N-substituting group comprises optional substituted C _1-4Alkyl, optional substituted C ₆Aryl and acyl group are (with C _1-4Alkyl is as acyl substituent).

Z

Z preferably takes the circumstances into consideration to be selected from S and O, more preferably S.

R ^N5And R ^N6

To R ^N5And R ^N6Preferably can be with above to R ^N3And R ^N4That explains is preferably identical.

R ^N1And R ^N2

In formula I compound, work as R ^N1And R ^N2When the nitrogen-atoms that connects with their formed the heterocycle with 4 to 8 atoms, this can form the C of the above definition that must contain at least one azo-cycle atom _4-20The part of heterocyclic radical (except that having minimum 4 annular atomses).Preferred R ^N1And R ^N2The nitrogen-atoms that connects with their form have 5,6 or 7 atoms, the more preferably heterocycle of 6 annular atomses.

Monocycle with a nitrogen-atoms comprises azetidine, azetidine, tetramethyleneimine (Pyrrolidine), pyrroline (for example 3-pyrroline, 2,5-pyrrolin), 2H-pyrroles or 3H-pyrroles (different pyrroles), piperidines, dihydropyridine, tetrahydropyridine and azepine

Monocycle with two nitrogen-atoms comprises imidazolidine, pyrazolidine (diazacyclo pentane), tetrahydroglyoxaline, pyrazoline (pyrazoline) and piperazine; Monocycle with a nitrogen and an oxygen comprises that Si Qing oxazole, dihydro-oxazole, tetrahydrochysene isoxazole, dihydro-isoxazole, morpholine, Si Qing oxazine, Er Qing oxazine are with oxazine; Monocycle with a nitrogen and a sulphur comprises thiazoline, thiazolidine and parathiazan.

Preferred ring be denitrogenate outer also contain one heteroatomic those, and especially, preferred heteroatoms is oxygen and sulphur.Therefore, preferred group comprises that morpholino base, thiomorpholine are for base, thiazolinyl.Do not contain other heteroatomic preferred group and comprise pyrrolidino.

Most preferred group is that morpholino base and parathiazan are for base.

As mentioned above, these heterocyclic groups can itself be substituted; A preferred class substituting group is C _1-7Alkyl.When heterocyclic group was the morpholino base, substituting group is methyl or ethyl preferably, and more preferably methyl.Unique methyl substituents is most preferably at 2.

Except that above listed monocyclic groups, bridge or crosslinked ring have also been considered to have.Wherein this group example of ring of containing these types of nitrogen and Sauerstoffatom has:

These be named as respectively 8-oxa--3-aza-bicyclo [3.2.1] oct-3-yl, 6-oxa--3-aza-bicyclo [3.1.0] oneself-3-base, 2-oxa--5-aza-bicyclo [2.2.1] heptan-5-base and 7-oxa--3-aza-bicyclo [4.1.0] heptan-the 3-base.

Universal synthesis method

R wherein ⁴Be Q-Y-X and Q be-NH-C (=O)-formula I compound can be expressed as formula 1:

Formula I

Wherein-Y-X is not C _1-7These compounds of alkyl can be by formula 2 compounds by preparing with suitable amine or thiol reactant:

Formula 2

Wherein L is a leavings group, for example chlorine or bromine.This reaction can be carried out in room temperature, perhaps can heat if necessary.

Formula 2 compounds can be synthetic by the reaction of formula 3 compounds and formula 4 compounds is come:

Formula 3

Formula 4

Wherein-Y-X is C _1-7Formula 1 compound of alkyl can be synthetic by the reaction of formula 3 compounds and formula 4a compound is come:

Formula 4a

Formula 3 compounds can be also synthetic originally with formula 5 compounds as containing zinc acetate by using suitable reductive agent:

Formula 5

Formula 5 compounds can through type 6 and the Suzuki-Miyaura coupling of 7 compound synthesize:

Formula 6

Formula 7

Wherein X ' is a group, for example bromine or OTf.The coupling part can be reversed.

Formula 7 compounds can followingly synthesize.

Formula 7a compound

Formula 7a

Can synthesize by cyclocondensation by formula 8a compound through pyrolysis, decarboxylation:

Formula 8a

Formula 8a compound can by formula 9a compound by with suitable formula HNR ^N1R ^N2Amine in The suitable solvent, react to come synthetic:

Formula 9a

Formula 9a compound can be by formula 10a compound

Formula 10a

In The suitable solvent, react synthetic by Meldrum acid derivative with formula 11a:

Formula 11a

Formula 7b compound

Formula 7b

Can be by making formula 8b compound

Formula 8b

In solvent such as DCM, in the presence of alkali such as triethylamine, react synthetic with trifluoromethanesulfanhydride anhydride.

Formula 8b compound can be by formula 9b compound by chlorine by formula HNR ¹R ²The amine nucleophilic substitution synthesize:

Formula 9b

Formula 9b compound can be by formula 10b compound by using chlorizating agent such as POCl ₃Carrying out chlorination synthesizes:

Formula 10b

Formula 10b compound can be synthesized by reacting next with diethyl malonate or its Equivalent by formula 11b compound:

Formula 11b

Formula 7c compound:

Formula 7c

The approach of formula 7c compound has description in WO 03/024949 (route of synthesis 6).

R wherein ⁴Be Q-Y-X, Q be-O-and X be selected from SR ^S1Or NR ^N3R ^N4Formula I compound can be expressed as formula 13:

Formula 13

X wherein " expression SR ^S1Or NR ^N3R ^N4These compounds can be by formula 14 compounds by synthesizing with suitable amine or thiol reactant:

Formula 14

Wherein L is a leavings group, for example chlorine or bromine.This reaction can be carried out in room temperature, perhaps can heat if necessary.

Formula 14 compounds can through type 15 compounds

Formula 15

With formula 16 compounds:

Formula 16

If wherein Y is asymmetric, then preferred L is not Br,

Leaving reaction at for example salt of wormwood comes synthetic.

Formula 15 compounds can use diazotization-method for hydrolysis to be synthesized by formula 3 compounds.This at first for example uses HBF ₄With butyl nitrite amino is converted into the diazonium fluoride borate, for example uses cupric oxide (I)-cupric nitrate (II) aqueous solution then its hydrolysis.

Wherein Q be-O-and X be-C (=O)-NR ^N5R ^N6Formula I compound can be expressed as formula 17:

Formula 17

X wherein " ' expression NR ^N5R ^N6These compounds can be by formula 18 compounds by react to synthesize in the presence of HBTU and HOBT with suitable amine:

Formula 18

Formula 18 compounds can be prepared by reacting in methyl alcohol with sodium hydroxide by formula 19 compounds:

Formula 19

Formula 19 compounds can come synthetic by leaving reaction with formula 20 compounds at for example salt of wormwood by formula 15 compounds:

Formula 20

R wherein ⁴The The compounds of this invention that is H can be by to be similar to preparing for boric acid and formula 7 compound couplings that above-mentioned method will suit.

The purposes of compound of the present invention

The invention provides active compound, specifically is active 8-aryl-2-amine-4-yl-quinoline-4-ketone, Pyridopyrimidine-4-ketone and chromene-4-ketone.

Term " activity " relates to and can suppress the active compound of DNA-PK as used herein, and specifically comprise the compound (medicine) with intrinsic activity and the prodrug of these compounds, described prodrug can itself demonstrate very little intrinsic activity or not have intrinsic activity.

A kind of assay method that can be used to estimate the DNA-PK inhibition of being bestowed by specific compound has description in following examples.

The present invention also provides the method that suppresses DNA-PK in cell, and this method comprises makes described cell contact with the active compound of significant quantity, and preferably the form with pharmaceutically acceptable composition contacts.This method can be implemented in external or body.

For example, can be at cultured cell in vitro (for example from tumour) sample, the material and the described cells contacting that make active compound and have known curative effect are observed the curative effect increase of compound to those cells.

The present invention also provides the active active compound of inhibition DNA-PK and has suppressed the active method of DNA-PK, and this method comprises makes cell contact in external or body with the active compound of significant quantity.

Active compound can also be as the cell cultures additive to suppress DNA-PK, for example in order to make cell external responsive to known chemotherapeutic or ionizing radiation treatment.

Active compound can also be as the part of external test method, for example whether may have benefited from the treatment carried out with in question compound in order to measure candidate host.

The present invention also provides the active compound of the method that is used for the treatment of human body or animal body.This method can comprise to the active compound of this curee's administering therapeutic significant quantity, preferably use with the form of pharmaceutical composition.

Be usually directed to wherein can to obtain certain required result of treatment, for example suppress human or animal's's (for example in the animal doctor uses) of illness development treatment as this paper used term " treatment " in sanatory context, this term comprises the stopping of reduction, tempo of tempo, the improvement of illness and the healing of illness.This term also comprises the treatment as preventive measures (i.e. prevention).

Term " treatment significant quantity " relates to active compound or comprises the amount that can produce certain required result of treatment effectively and match with rational interests/risk ratio of material, composition or the formulation of active compound as used herein.

Term " adjuvant " relates to active compound and known methods of treatment is united use as used herein.These methods comprise as being used for the treatment of the cytotoxic drug scheme and/or the ionizing radiation of various cancers type.Can include but not limited to following with the example of the auxiliary carcinostatic agent that makes up from compound of the present invention: alkylating agent: mustargen, chlormethine (mechlorethamine), endoxan, ifosfamide, melphalan, Chlorambucil; Nitrosourea: carmustine (BCNU), Luo Mositing (CCNU), semustine (Semustine), ethyleneimine/first melamine, Tris-s-triazine (TEM), triethylene thiophosphoramide (plug is for group), altretamine (HMM); Alkylsulfonate/ester, busulfan; Triazines, Dacarbazine (dacarbazine) is (DTIC); Metabolic antagonist; Folacin, methotrexate, trimetrexate, pyrimidine analogue, 5 FU 5 fluorouracil, fluorodeoxyuridine, gemcitabine, cytosine arabinoside (AraC), 5-azacytidine, 2,2 '-difluoro Deoxyribose cytidine; Purine analogue; 6-mercaptopurine, 6-Tioguanine, azathioprine, 2 '-deoxycoformycin (pentostatin, erythrohydroxynonyladenine (EHNA), fludarabine phosphate, 2-chlorodeoxyadenosine (CldAdo, 2-CdA); The topoisomerase I inhibitor; Camptothecine, Hycamtin, irinotecan, rubitecan (rubitecan); Natural product; Antimitotic drug, taxol, vinca alkaloids, vincaleucoblastine (VLB), vincristin, vinorelbine, Taxotere ^TM(docetaxel), estramustine, phosphoric acid estramustine; Epipodophyllotoxin, Etoposide, teniposide; Microbiotic; Dactinomycin (actimomycin D), Rubomycin C (daunorubicin), Zorubicin (Dx), mitoxantrone, idarubicin, bleomycin, Plicamycin (Plicamycin), ametycin, dactinomycin; Enzyme; L-Asparaginase, RNA enzyme A: biological response modifiers; Interferon-' alpha ', IL-2, G-CSF, GM-CSF; Differentiation agent; Retinoic acid derivatives; Radiosensitizer; Metronidazole, Misonidazole, demethyl Misonidazole, a hair Buddhist nun azoles, etanidazole, Nimorazole, RSU 1069, EO9, RB 6145, SR4233, niacinamide, 5-bromouracil deoxyribose, idoxuridine, bromine Deoxyribose cytidine; Platinum coordination complex; Cis-platinum, carboplatin: amerantrone; Urea, hydroxyurea that mitoxantrone, AQ4N replace; Methylhydrazine derivative, N-methylhydrazine (MIH), Procarbazine; Adrenal cortex inhibitor, mitotane (o.p '-DDD), aminoglutethimide; Cytokine; Interferon, rabbit (α, β, γ), interleukin-; Hormone and antagonist; Adrenocortical steroid/antagonist, prednisone and Equivalent, dexamethasone, aminoglutethimide; Progesterone, Hydroxyprogesterone caproate bp 98, medroxyprogesterone acetate, Magace; Oestrogenic hormon, stilboestrol, lynoral/Equivalent; Estrogen antagonist material, tamoxifen; Male sex hormone, testosterone propionate, FL/Equivalent; Antiandrogen, flutamide, gonadotropin releasing hormone analogues, leuprorelin acetate; Non-steroid antiandrogen, flutamide; EGFR inhibitor, VEGF inhibitor; Proteasome inhibitor.

Cancer

The invention provides the carcinostatic agent that is used for the treatment of cancer or the active compound of adjuvant.Those of ordinary skills can determine easily whether candidate compound can treat the carcinous illness of the cell type of any specific alone or in combination.

The example of cancer includes but not limited to lung cancer, small cell lung cancer, gastrointestinal cancer, intestinal cancer, colorectal carcinoma, breast cancer, ovarian cancer, prostate cancer, carcinoma of testis, liver cancer, kidney, bladder cancer, pancreas cancer, the cancer of the brain, sarcoma, osteosarcoma, Kaposi sarcoma, melanoma and leukemia.

The cell of any type be can treat, lung, stomach and intestine (for example comprising intestines, colon), breast, ovary, prostate gland, liver, kidney, bladder, pancreas, brain and skin included but not limited to.

More than Ding Yi anticancer therapy can be used as single therapy and uses, and perhaps, except that compound of the present invention, can also comprise conventional operation or radiotherapy or chemotherapy.This chemotherapy can comprise the antineoplastic agent of one or more following kinds:

(i) other antiproliferative agents/antitumor formation medicine and the combination thereof as being used for the medical science oncology, for example alkylating agent (for example cis-platinum, oxaliplatin, carboplatin, endoxan, mustargen, melphalan, Chlorambucil, busulfan, Temozolomide and nitrosourea); Metabolic antagonist (for example gemcitabine and antifol, for example the fluorine pyrimidine is as 5 Fluracils and Tegafur, Raltitrexed, methotrexate, cytosine arabinoside and hydroxyurea); Antitumor antibiotics (for example anthracycline antibiotics, as Dx, bleomycin, Zorubicin, daunorubicin, epirubicin, idarubicin, Mitomycin-C, dactinomycin and Plicamycin); Anti-fissuring agent (for example vinca alkaloids such as vincristin, vincaleucoblastine, vindesine and vinorelbine and Taxan such as taxol and taxotere (taxotere) and polokinase inhibitor); And topoisomerase enzyme inhibitor (for example epipodophyllotoxin such as Etoposide and teniposide, amsacrine, Hycamtin and camptothecine);

(ii) cytostatics, for example estrogen antagonist material (for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene), antiandrogen (for example bicalutamide, flutamide, Nilutamide and cyproterone acetate), lhrh antagonist or LHRH agonist (for example goserelin, Leuprolide and buserelin), progestogen (for example Magace), aromatase inhibitor (for example Anastrozole, letrozole, vorazole and Exemestane) and 5 ^*-reductase inhibitor such as finasteride;

(iii) anti-invasion agent (c-Src kinases man group inhibitor for example is as 4-(6-chloro-2,3-methylene dioxo group aniline base)-7-[2-(4-methylpiperazine-1-yl) oxyethyl group]-5-tetrahydropyran-4-base oxygen base quinazoline (AZD0530; International Patent Application WO 01/94341) and N-(2-chloro-6-aminomethyl phenyl)-2-{6-[4-(2-hydroxyethyl) piperazine-1-yl]-2-methylpyrimidine-4-base is amino thiazole-5-methane amide (dasatinib, BMS-354825; J.Med.Chem., 2004,47,6658-6661), and the antibody of inhibitors of metalloproteinase such as marimastat, UPA function of receptors inhibitor or heparinase (Heparanase));

(iv) somatomedin depressant of functions: for example these inhibitor comprise growth factor antibodies and growth factor receptor antibody (for example anti-erbB 2 antibody Si Tuman cloth [HerceptinT], anti-EGFR-antibodies panitumumab, anti-erbB1 antibody Cetuximab [cetuximab/Erbitux, C225] and by people such as Stern (Critical reviews in oncology/haematology, 2005, the 54th volume, the 11-29 page or leaf) disclosed any somatomedin or growth factor receptor antibody); These inhibitor also comprise tyrosine kinase inhibitor, epidermal growth factor family inhibitor (EGFR family tyrosine kinase inhibitor for example for example, as N-(3-chloro-4-fluorophenyl)-7-methoxyl group-6-(3-morpholino propoxy-) quinazoline-4-amine (Gefitinib (gefitinib), ZD1839), N-(3-ethynyl phenyl)-6, two (2-methoxyethoxy) quinazolines of 7--4-amine (erlotinib (erlotinib), OSI 774) and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholino propoxy-)-quinazoline-4-amine (CI 1033); The erbB2 tyrosine kinase inhibitor is as lapatinib; PHGF man group inhibitor; Platelet-derived growth factor family inhibitor is as imatinib; Serine/threonine kinase inhibitor (for example the Ras/Raf signal sends inhibitor, and farnesyl transferase inhibitor for example is as Xarelto (sorafenib) (BAY 43-9006)); Send inhibitor by MEK and/or the kinase whose cell signal of AKT; PHGF man group inhibitor; The c-kit inhibitor; The abl kinase inhibitor; IGF acceptor (rhIGF-1) kinase inhibitor; Aurora kinase inhibitor (for example AZD1152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 and AX39459); And cell cycle protein dependent kinase inhibitor, as CDK2 and/or CDK4 inhibitor;

(v) anti-angiogenic agent, for example suppress those of vascular endothelial growth factor effect, [anti-vascular endothelial cell growth factor antibody rhuMAb-VEGF (AvastinT) and vegf receptor tyrosine kinase inhibitor such as 4-(4-bromo-2-fluoroanilino)-6-methoxyl group-7-(1-methyl piperidine-4-ylmethoxy) quinazoline (ZD6474 for example; Embodiment 2 among the WO 01/32651), 4-(4-fluoro-2 methyl indole-5-base oxygen base)-6-methoxyl group-7-(3-tetramethyleneimine-1-base propoxy-) quinazoline (AZD2171; Embodiment 240 among the WO 00/47212), vatalanib (PTK787; WO 98/35985) and SU11248 (sunitinib; WO01/60814), for example those disclosed compound and the compound (for example linomide (linomide), integral protein avb3 depressant of functions and angiostatin) that plays a role by other mechanism in International Patent Application WO 97/22596, WO 97/30035, WO 97/32856 and WO 98/13354];

(vi) blood vessel injury agent, for example combretastatin (Combretastatin) A4 and in International Patent Application WO 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO 02/08213 disclosed compound;

(vii) antisense therapy for example is oriented to those of above listed target position, and for example ISIS 2503, a kind of anti--the ras antisense drug;

(viii) gene therapy method for example comprises the method for replacing aberrant gene such as unusual p53 or unusual BRCA1 or BRCA2; The GDEPT enzyme prodrug of the gene orientation (treatment) method is for example used those of Isocytosine deaminase, thymidine kinase or bacterium nitroreductase; And the increase patient treats the method such as the multidrug resistance gene of chemotherapy or radiotherapy tolerance; And

(ix) immunotherapy method, for example comprise increase the patient tumors cell immunogenic in vitro with body in method, for example carry out transfection with cytokine such as interleukin II, interleukin-4 or rHuGM-CSF; Reduce the method for T cell anergy; The method of the immunocyte of use transfection such as the dendritic cell of cytokine transfection; Use the method for the tumor cell line of cytokine transfection; And use anti-spy to answer the method for antibody.

Use

Active compound or comprise the pharmaceutical composition of active compound can be by any route of administration general/periphery ground or be applied to the curee easily at the position of required effect, described route of administration includes but not limited to: oral (for example by eating); Local (for example comprise in transdermal, the nose, eye, oral cavity and hypogloeeis); Lung (for example, for example using aerosol, for example through port or nose) by sucking or being blown into treatment; Rectum; Vagina; Outside the gi tract,, comprise in subcutaneous, intracutaneous, intramuscular, intravenously, intra-arterial, intracardiac, the sheath, in the vertebra, in the capsule, under the capsule, in the socket of the eye for example by injection, in the intraperitoneal, tracheae, under the epidermis, intraarticular, arachnoid membrane be down and in the breastbone; Implant depot formulation by for example subcutaneous or intramuscular.

The curee can be eukaryote, animal, vertebrates, Mammals, rodent (for example cavy, hamster, rat, mouse), murine (for example mouse), Canis animals (for example dog), feline (for example cat), equine species (for example horse), primate, ape (for example monkey or ape), monkey (for example marmoset monkey, baboon), ape (for example gorilla, chimpanzee, orangutan, gibbon) or people.

Preparation

Although active compound might be used separately, but preferably present as its pharmaceutical composition (for example preparation), said composition comprises at least a active compound as defined above and one or more pharmaceutically useful carriers, adjuvant, vehicle, thinner, weighting agent, buffer reagent, stablizer, sanitas, lubricant or other materials well known to those skilled in the art, randomly with other treatment agent or preventive.

Thereby, the present invention further provides the method for pharmaceutical composition and pharmaceutical compositions as defined above, comprise at least a active compound as defined above and one or more pharmaceutically useful carrier, vehicle, buffer reagent, adjuvant, stablizer or other materials as described herein are mixed.

Term " pharmaceutically useful " relates to and is applicable within the scope of rational medical judgment and contacts with curee's (for example people) tissue and do not have over-drastic toxicity, pungency, anaphylaxis or other problem or complication and the compound that matches with rational interests/risk ratio, material, composition and/or a formulation as used herein.Every kind of carrier, vehicle etc. can must also be " acceptable " aspect other preparation components compatibility.

Suitable carrier, vehicle etc. can be at the medicine textbooks of standard, for example Remington ' s Pharmaceutical SciencesFind in (the 18th edition, Mack publishing company, Easton, Pa., 1990).

Preparation can be unit dosage aptly, can be prepared by the method that pharmaceutical field is known arbitrarily.These class methods comprise makes active compound and the carrier step linked together that constitutes one or more attachment components.Generally speaking, preparation is prepared as follows: make solid carrier or this two all even closely associating of active compound and liquid vehicle or fine pulverizing, then if necessary with the product moulding.

Preparation can be the form of liquid, solution, suspension, emulsion, elixir, syrup, tablet, lozenge, granule, pulvis, capsule, cachet, pill, ampulla, suppository, vaginal suppository, ointment, gelifying agent, paste, creme, sprays, mist agent (mist), foaming agent, lotion, finish, bolus, electuary or aerosol.

The preparation that is suitable for Orally administered (for example by eating) can be discrete unit, for example capsule, cachet or tablet, every dose of active compound that contains predetermined amount; Pulvis or granule; Solution in water-based or non-aqueous liquid or suspension; Oil-in-water liq emulsion or water-in-oil-type liquid emulsion; Bolus; Electuary; Or paste.

Tablet can prepare by conventional means, and for example compacting or molded is randomly together with one or more attachment components.Compressed tablet can be prepared as follows: free-flowing form such as the powder or the particle of compacting active compound in the machinery that is fit to randomly are mixed with one or more tackiness agents (polyvidone for example, gelatin, gum arabic, Sorbitol Powder, tragacanth gum, Vltra tears), weighting agent or thinner (lactose for example, Microcrystalline Cellulose, secondary calcium phosphate), lubricant (Magnesium Stearate for example, talcum, silicon-dioxide), disintegrating agent (sodium starch glycolate for example, polyvinylpolypyrrolidone, croscarmellose sodium), tensio-active agent or dispersion agent or wetting agent (for example Sodium Lauryl Sulphate BP/USP) and sanitas are (for example right-methyl hydroxybenzoate, right-nipasol, Sorbic Acid).Molded tablet can be prepared as follows: in the machinery that is fit to molded with inert liquid diluent moistening the mixture of powder compound.Tablet can be randomly by dressing or delineation, and can provide the wherein slowly-releasing or the controlled release of active compound through preparation, for example uses the Vltra tears of different ratios, so that required release profiles to be provided.Tablet can randomly have enteric coating, to be provided at intestines parts but not the release in the stomach.

The preparation that is suitable for topical (for example in the transdermal, nose, eye, cheek and hypogloeeis) can be formulated into ointment, creme, suspension, lotion, pulvis, solution, paste, gelifying agent, sprays, aerosol or finish.Select as an alternative, preparation can comprise medicine and paste or dressing, for example is impregnated with active compound and one or more vehicle chosen wantonly or the bandage or the binding property plaster of thinner.

The preparation that is suitable for mouthful interior topical application comprises lozenge, through the matrix of flavoring, be generally in sucrose and gum arabic or the tragacanth gum and comprise active compound; Pastille (pastille) comprises active compound in inert base such as gelatin and glycerine or sucrose and gum arabic; And mouth wash shua, in the liquid vehicle that is fit to, comprise active compound.

Be suitable for the preparation of eyes topical application is also comprised eye drops that wherein active compound is dissolved or suspended in the carrier, the especially aqueous solvent that are fit to that is used for active compound.

Wherein carrier is that the preparation that solid is suitable for nasal administration comprises meal, and particle diameter for example about 20 to about 500 microns scope, is taked the mode administration of snuffing, that is to say from be close to the dust container that nose places to suck rapidly by the nostril.Wherein carrier is to be used for that for example nasal spray, nasal drop are used or the suitable preparation of the liquid by the spraying gun aerosol administration, comprises the aqueous solution or the oil solution of active compound.

Those that the arosol spray of self-pressurization packing is presented since being suitable for sucking the preparation of using and comprising wherein adopt the propelling agent that is fit to, for example Refrigerant 12, trichlorine methyl fuoride, dichloro tetrafluoro ethane, carbonic acid gas or other gas that is fit to.

Be suitable for comprising ointment, creme and emulsion via the preparation of topical application.In the time of in being formulated in ointment, active compound can randomly be used with paraffin class or water-miscible ointment base.Select as an alternative, can utilize the oil-in-water-type cream base that active compound is formulated in the creme.If desired, the water of cream base for example can comprise at least about 30%w/w polyvalent alcohol, promptly has the alcohol of two or more hydroxyls, for example propylene glycol, butane-1,3-glycol, mannitol, Sorbitol Powder, glycerine and polyoxyethylene glycol and composition thereof.Topical formulations may need to comprise that the enhanced activity compound passes skin or other affected area absorb or the compound of infiltration.The example of this class skin penetration enhancer comprises dimethyl sulfoxide (DMSO) and relevant analogue.

When being mixed with local emulsion, oil phase can randomly only comprise emulsifying agent, perhaps it can comprise at least a emulsifying agent with fat or oil or with fat and oil mixture.Preferably, comprise hydrophilic emulsifier and the lipophilic emulsifier that serves as stablizer.Also preferably comprise oil ﹠ fat simultaneously.One or more emulsifying agents, choose any one kind of them or multiple stablizer constitutes so-called emulsifying property wax, this wax constitutes so-called emulsifying property ointment base with oil and/or fat, and the latter constitutes the oily disperse phase of creme.

The emulsifying agent and the emulsion stablizer that are fit to comprise polysorbate60, sorbester p17, cetostearyl alcohol, tetradecyl alcohol, Zerol and Sodium Lauryl Sulphate BP/USP.Oily or the fatty selection that is suitable for preparation is based on realizing required beauty treatment character, because the solubleness of active compound in most of oil that may be used in the pharmaceutical emulsion may be very low.Thereby, creme preferably non-greasy, do not fade and rinsable product, have suitable viscosity, from pipe or other containers, spill avoiding.Can use straight or branched monobasic or binary hydrocarbyl carbonate, for example propylene glycol diesters, Isopropyl myristate, decyl oleate, Wickenol 111, butyl stearate, the palmitinic acid 2-(ethyl hexyl) ester of dissident's two acid diesters, the different hexadecyl ester of stearic acid, coconut fatty acid or be called the branched ester adulterant of Crodamol CAP, back three is preferred ester.They can separately or unite use, and this depends on required character.

Select as an alternative, can use the high-melting-point lipid, for example soft white paraffin and/or whiteruss or other mineral oil.

The preparation that is suitable for rectal administration can be suppository, has suitable matrix and for example comprises theobroma oil or salicylate.

The preparation that is suitable for vaginal application can present vaginal suppository, tampon, creme, gelifying agent, paste, foaming agent or sprays, and it also contains appropriate carriers known in the art except active compound.

Be suitable for parenteral administration (for example injection, comprise skin, subcutaneous, intramuscular, intravenously and intradermal) preparation comprise water-based and non-aqueous isoosmotic, pyrogen-free, aseptic injectable solution, it can contain antioxidant, buffer reagent, sanitas, stablizer, bacteriostatic agent and make the preparation and the isoosmotic solute of person's blood that is subjected to the medicine; Water-based and non-aqueous sterile suspension, it can comprise suspension agent and thickening material; Make compound be oriented to liposome or other microparticulate systems of blood constitutent or one or more organs with being designed to.Being suitable for use in grade in this class preparation oozes the example of carrier and comprises sodium chloride injection, Ringer's solution or lactated ringer's inj.Usually, the concentration of active compound in solution is extremely about 10 μ g/ml of about 1ng/ml, and for example about 10ng/ml is to about 1 μ g/ml.Preparation can be present in single agent or the multi-agent sealed vessel, for example ampoule and bottle, and can be stored under lyophilize (freeze-drying) condition, only need to add sterile liquid carrier before use soon and get final product, for example water for injection.Can prepare interim injection solution and suspension from sterile powder, granule and tablet.Preparation can be to be designed to make active compound to be oriented to the form of liposome or other microparticulate systems of blood constitutent or one or more organs.

Dosage

Be understandable that active compound can be different because of the patient with the suitable dosage that comprises the composition of active compound.Determine that optimal dose generally will involve the treatment benefit level of the present invention's treatment and the balance between any risk or the harmful side effect.Selected dosage level will depend on multiple factor, include but not limited to activity, route of administration, time of application, the discharge rate of compound, the time length of treatment, the other drug of uniting use, compound and/or material and patient's age, sex, body weight, condition, general health situation and the medical history of specific compound.The amount and the route of administration of compound depend on the doctor the most at last, but generally speaking, dosage will be realized required effect at the partial concn that site of action obtains, and do not cause substantive injury or harmful side effect.

Vivo medicine-feeding can carry out with potion, and continuously or off and on (for example by the dosage of appropriate intervals to separate) spreads all over therapeutic process.Determine that the most effective method of using means and dosage is well-known to those skilled in the art, will be different because of the used preparation of therapy, therapeutic purpose, the target cell of being treated and the curee who is treated.By the gentle mode of attending doctor's selective agent water gaging, can carry out single or multiple and use.

Usually, the appropriate dosage scope of active compound is the body weight/day of about 100 μ g to about 250mg/ kilogram curee.When active compound was salt, ester, prodrug etc., the amount of being used was that calculate on the basis with the parent compound, and therefore used actual weight should increase pro rata.

Embodiment

Following examples only are not intended to limit scope of the present invention as described herein for the present invention is described.

Acronym

For simplicity; use well-known abbreviation to represent the number of chemical part, include but not limited to methyl (Me), ethyl (Et), n-propyl (nPr), sec.-propyl (iPr), normal-butyl (nBu), the tertiary butyl (tBu), n-hexyl (nHex), cyclohexyl (cHex), phenyl (Ph), xenyl (biPh), benzyl (Bn), naphthyl (naph), methoxyl group (MeO), oxyethyl group (EtO), benzoyl (Bz) and ethanoyl (Ac).

For simplicity, use well-known abbreviation to represent the number of chemical compound, include but not limited to methyl alcohol (MeOH), ethanol (EtOH), Virahol (i-PrOH), methyl ethyl ketone (MEK), ether (Et ₂O), acetate (AcOH), methylene dichloride (DCM), trifluoroacetic acid (TFA), dimethyl formamide (DMF), tetrahydrofuran (THF) (THF) and dimethyl sulfoxide (DMSO) (DMSO).

General experiment is described in detail

Chemical is available from Aldrich chemical company, the synthetic company limited of Lancaster and AcrosOrganics (Fisher Scientific Britain company limited).THF is fresh distillation from sodium/benzophenone.Methyl alcohol and ethanol distill from magnesium/iodine.DCM carries out drying through the Vanadium Pentoxide in FLAKES distillation.Acetone carries out drying through the hydrolith distillation.All solvents that do not use are immediately used molecular sieve (4 under nitrogen

The 3-5mm pearl) preserves.Dry DMF is by the SureSeal of Aldrich ^TMObtain in the bottle.Triethylamine is carried out drying through the hydrolith distillation, and under nitrogen, preserve with potassium hydroxide.

Use the Merck silica gel 60F that is coated in advance on the aluminium sheet ₂₅₄Carry out tlc (TLC),, use shortwave (254nm) ultraviolet ray or by with triketohydrindene hydrate or vitriolization, handle with Vanillin and carry out video picture then subsequently with the aluminium sheet drying.Use Davisil silica gel (40-63 μ m) under middle pressure, to carry out ' fast ' column chromatography.

Use Stuart Scientific SMP3 Instrument measuring fusing point, do not proofread and correct.Use BrukerSpectrospin AC 300E spectrometer ( ¹H 300MHz or ¹³C 75MHz) or Bruker SpectrospinAC 500E spectrometer ( ¹H 500MHz or ¹³C 125MHz) obtains ¹H and ¹³C nucleus magnetic resonance (NMR) spectrum.Use the residual solvent peak as interior mark, with PPM (δ) the report chemical shift of low of tetramethyl-sulfone (teramethylsulfone).Make up by s (unimodal), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad peak) or its and to show multiplicity.Use Micromass Platform instrument to obtain the LC/MS spectrum with positively charged ion or the operation of anionic electrodeposition spray patterns.Use C18 post (50 * 4.6mm; Supelco Discovery or Waters Symmetry) realize separating with 15 minutes 0.05% formic acid and the gradient elution of methyl alcohol (10-90%).On Bio-Rad FTS 3000MX diamond ATR, compose with pure sample product record IR.

Synthesizing of key intermediate

(i) three fluoro-methylsulfonic acid 1-nitro-dibenzothiophene-4-base ester (6)

(a) dibenzothiophene-4-alcohol (i)

To the cooling (78 ℃) dibenzothiophene (20.8g, (1.7M is in pentane 113mmol) to add tert-butyl lithium in the solution in anhydrous THF (400ml); 100ml, 170mmol).In-78 ℃ reaction mixture was stirred 1 hour, make it be warmed to room temperature then, stirred like this 16 hours.Mixture is cooled to 0 ℃ then, (1M is in THF with ethyl-magnesium-bromide with slug flow by conduit; 170ml 170mmol) joins in the amber reaction mixture.Make this reactant once more to room temperature, like this it was stirred 30 minutes thereupon.Reflux exchanger is connected with reactor, then oxygen was passed through this solution bubbling 40 minutes.Then this reactant was stirred 1 hour in addition, then it is inclined carefully to trash ice, be acidified to pH3 with dense HCl.Use ethyl acetate (3 * 80ml) extraction mixtures then.Then organic extract liquid is handled with the 3M sodium hydroxide solution, until reaching pH10.Separate this alkalescence water layer, be acidified to pH3, cause the oily solid precipitation with 2M HCl.It is dissolved in the ether (150ml), uses MgSO ₄Drying is filtered, vacuum concentration, then from ethanol: water (1: 1) (250ml) recrystallization, obtain being equivalent to the beige solid (21.6g, 96%) of title compound, it need not to be further purified.M/z (LC-MS, ESP), RT=3.64 minute, (M+H)=201.1.

(b) 4-methoxyl group-dibenzothiophene (ii)

To dibenzothiophene-4-alcohol (i) (14.2g, 71.0mmol) add in the solution in acetone (500ml) potassium carbonate powder (14.72g, 106.5mmol) and methyl-iodide (4.43ml, 71mmol).This mixture heating up to refluxing, was stirred 16 hours like this.With this mixture cooling, pass through Celite then ^TMPad filters.With gained filtrate vacuum concentration, obtain the oily resistates, it with methylene dichloride (100ml) dilution, is washed with 1M NaOH and saturated brine solution.Use MgSO ₄With the organic layer drying, filter, vacuum concentration obtains being equivalent to the beige solid of title compound, and it uses without any being further purified.(15.2g, 100%) m/z (LC-MS, ESP), RT=4.22 minute, (M+H)=215.1.

(c) the 4-methoxyl group-1-nitro-dibenzothiophene (iii)

(ii) (4.3g 20.0mmol) is dissolved in the Glacial acetic acid (60ml), adds nitrosonitric acid (3.37ml) in dropwise mode in this solution, is no more than 25 ℃ with the temperature of guaranteeing this mixture with 4-methoxyl group-dibenzothiophene.Should stir 45 minutes in addition by the yellow suspension, then it is inclined carefully to water (200ml), stirred 15 minutes.By removing by filter yellow solid, with big water gaging, use a large amount of hexane thorough washing then.The resistates that will obtain thus is dry in vacuum drying oven then, obtains yellow solid-state title compound, and it uses without any being further purified.(5.19g, 97%) m/z (LC-MS, ESP), RT=4.15 minute, (M+H)=260.1.

(d) 1-nitro-dibenzothiophene-4-alcohol (iv)

With solid-state pyridine hydrochloride (1kg, 8.7mol) join 4-methoxyl group-1-nitro-dibenzothiophene (iii) (35.44g, 187mmol) in, with this reactant thorough mixing, be heated to 165 ℃ then, continue to stir.This mixture was kept 8 hours like this, cooling, water (500ml) dilution is with methylene dichloride (3 * 200ml) extractions.The 3M sodium hydroxide solution is joined in this organic extract liquid, until from solution, being settled out dark-coloured solid.Shift out filtrate, use dense HCl that liquid is acidified to pH1.By removing by filter the gained aureus solid that forms after the acidifying, it is washed with water then, drying obtains suitable pure title compound, and it uses without any being further purified.(35.44g, 77%) m/z (LC-MS, ESP), RT=3.69 minute, (M+H)=246.2, (M-H)=244.1.

(e) three fluoro-methylsulfonic acid 1-nitro-dibenzothiophene-4-base ester (6)

(iv) (5.37g, (9.20ml 66.00mmol), dissolves this suspension fully 22.0mmol) to add triethylamine in the suspension in methylene dichloride (75ml) to refrigerative (5 ℃) 1-nitro-dibenzothiophene-4-alcohol.Then with dropwise mode by syringe in this mixture, add trifluoromethanesulfanhydride anhydride (5.85ml, 33.00mmol).In this temperature this mixture was stirred 1 hour, incline to trash ice then.Melt the ice, use CH ₂Cl ₂(3 * 20ml) with this mixture extraction.Then with the organic layer drying (MgSO that merges ₄), filter, vacuum concentration obtains light amber oily matter, with it by silicon-dioxide pad wash-out (pure CH ₂Cl ₂), obtaining the title compound of suitable pure form, it uses without any being further purified.(8.30g, 99%) m/z (LC-MS, ESP), RT=4.40 minute, unionization.

(f) 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls) dibenzothiophene (7)

Three fluoro-methylsulfonic acid 1-nitro-dibenzothiophene-4-base ester (6) (250mg packs in clean exsiccant flask under argon gas, 0.66mmol), two (tetramethyl ethylene ketone) two boron (bis (pinacolato) diboron) (185mg, 0.73mmol), potassium acetate (390mg, 3.98mmol), PdCl ₂(two (triphen phosphino-) ferrocene) (27mg, 0.033mmol) and two (triphen phosphino-) ferrocene (19mg, 0.033mmol).Under vacuum,, use argon cleaning 3 times with this flask emptying.Add diox (20ml), this reaction mixture was stirred 12 hours in 90 ℃.With this reaction mixture with DCM (100ml) dilution, with the organic layer water (3 * 30ml), salt solution (1 * 30ml) continuous washing, dry (Na ₂SO ₄), vacuum evaporating solvent obtains nitro boric acid ester (7), and it uses without being further purified.

(ii) 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls) diphenylene-oxide (12)

(a) diphenylene-oxide-4-alcohol (7)

(120ml, (27.4g is 163mmol) in the solution in anhydrous THF 300mmol) to join diphenylene-oxide with n-BuLi in-78 ℃.In 40 ℃ this reactant is slowly heated, stirred 18 hours.Then in-5 ℃ with the cooling of this reactant, dropwise add MeMgBr (100ml, 300mmol).After adding is finished, this reactant was stirred 1 hour in room temperature.Load onto the reflux exchanger that has bubbler, oxygen was passed through this reactant bubbling 4 hours, in 40 ℃ this reactant is slowly heated during this period.Continue to blast oxygen and can not increase reaction process.By this reaction mixture is inclined carefully to ice in and with the reactant cancellation.By adding dense HCl pH is transferred to 3, product is extracted among the DCM.Resistates is carried out purifying by flash chromatography, use DCM/EP (6: 4) as eluent.After the evaporation, some unreacted diphenylene-oxide (13g, 47%) have also been separated.Obtain the product (10.9g, 59mmol, 36%) of white solid state: R _f=0.18 (DCM-EP 6: 4); Mp:102 ℃; λ _Max(EtOH)/nm 234; IR (cm ^-1) 3258,3049,1635,1603,1477,1436,1347,1309,1245,1189,1158; ¹H NMR, (300MHz, CDCl ₃) δ 5.40 (1H, s, OH), 7.05 (1H, d, J _H2-H3=8Hz, H-3), 7.27 (1H, d, J=8Hz), 7.38 (1H, t, J=7Hz), 7.47-7.56 (2H, m), 7.61 (1H, d, J=8Hz), 7.97 (1H, d, J=8Hz); ¹³C NMR, (75MHz, CDCl ₃) δ 111.84,112.79,113.95,121.00,122.97,123.72,124.63 (Cq), 125.90 (Cq), 127.26,141.35 (Cq), 144.31 (Cq), 156.04 (Cq); MS (EI) m/z 184.0 M ⁺HRMSC ₁₂H ₈O ₂[M+H] ⁺, calculated value: 184.0519, measured value: 184.0517.

(b) 4-methoxyl group-diphenylene-oxide (8)

With salt of wormwood (1.4g, 10.11mmol) and methyl-iodide (0.42ml, (1.24g is 6.74mmol) in the solution in acetone (65ml) 6.74mmol) to join diphenylene-oxide-4-alcohol.With this reactant reflux, stirred 18 hours.After the cooling, with this reaction mixture 1M sodium hydroxide, water and salt solution continuous washing.Organic layer through dried over mgso, is filtered, concentrate then, obtain the product (1.33g, 6.74mmol, 100%) of white needles, it uses without being further purified.R _f=0.37 (AcOEt-EP1: 19); Mp:49-50 ℃; λ _Max(EtOH)/nm 279; IR (cm ^-1) 3054,2839,1900,1633,1496,1451,1423,1330,1309,1267,1188,1089,931,893,782,737; ¹H NMR, (300MHz, CDCl ₃) δ 4.03 (3H, s, OCH ₃), 6.94 (1H, d, J _H2-H3=8Hz, H-3), 7.26 (1H, t, J=8Hz), 7.36 (1H, t, J=7Hz), 7.48 (1H, t, J=7Hz), 7.54 (1H, d, J=7Hz), 7.69 (1H, d, J=8Hz), 7.94 (1H, d, J=8Hz); ¹³C NMR, (75MHz, CDCl ₃) δ 56.11 (OCH ₃), 109.31,111.96,112.82,120.85,122.86,123.46,124.43 (Cq), 125.72 (Cq), 127.21,145.20 (Cq), 145.63 (Cq), 156.04 (Cq); MS (EI) m/z199.1M ⁺HRMS C ₁₃H ₁₀O ₂[M+H] ⁺, calculated value: 199.0754, measured value: 199.0754.

(c) 4-methoxyl group-1-nitro-diphenylene-oxide (9)

To 4-methoxyl group-diphenylene-oxide (3.15g; 15.89mmol) dropwise add nitrosonitric acid (2.6ml in the solution in Glacial acetic acid (50ml); 63.50mmol).In adition process, this reactant is remained on 20 ℃, stirred 3 hours.After finishing, this reaction mixture is inclined carefully to frozen water; By adding 1M sodium hydroxide pH is transferred to 7, product is extracted with DCM.Organic layer through dried over mgso, is filtered, concentrate then.Resistates is carried out purifying by flash chromatography, use ethyl acetate/EP (1: 19) as eluent.First kind of compound of wash-out is 4-methoxyl group-3-nitro-diphenylene-oxide (270mg, 1.11mmol, 7%) from the post, obtains the solid-state title compound of emulsus (1.85g, 7.62mmol, 48%): R then _f=0.13 (EtOAc-EP 1: 19); Mp:155-156 ℃; λ _Max(EtOH)/nm 239; IR (cm ^-1) 3092,2917,2851,2042,1876,1630,1568,1506,1438,1396,1342,1297,1274,1239,1205,1159,1129,1091,1002,938,888,831,812,738,676; ¹H NMR, (300MHz, CDCl ₃) δ 4.06 (3H, s, OCH ₃), 6.92 (1H, d, J=8Hz), 7.18 (1H, t, J=8Hz), 7.36 (2H, m), 8.17 (1H, d, J=8Hz), 8.63 (1H, d, J=8Hz); ¹³C NMR, (75MHz, CDCl ₃) δ 56.81 (OCH ₃), 107.53,111.81,120.50 (Cq), 121.14 (Cq), 122.27,123.71,126.32,129.55,136.29 (Cq), 145.20 (Cq), 150.53 (Cq), 157.02 (Cq); MS (EI) m/z 243.1 M ⁺HRMSC ₁₃H ₉NO ₄[M+H] ⁺, calculated value: 243.0526, measured value: 243.0528.

(d) 1-nitro-diphenylene-oxide-4-alcohol (10)

(2g 8.22mmol) heated 18 hours in pyridine hydrochloride (17g) with 4-methoxyl group-1-nitro-diphenylene-oxide in 150 ℃.Make this reactant be cooled to 90 ℃, add 20ml water.After the cooling, product is extracted with DCM.Organic layer through dried over mgso, is filtered, concentrate then.Resistates is carried out purifying by flash chromatography, use DCM as eluent.Obtain yellow solid-state product (1.88g, 8.22mmol, 100%): R _f=0.13 (AcOEt-EP 1: 4); Mp:175 ℃; λ _Max(EtOH)/nm239; IR (cm ^-1) 1626,1577,1487,1442,1273,1240,1197,1153,1076,1016,983,912,817.; ¹H NMR, (300MHz, CDCl ₃) δ 6.35 (1H, s, OH), 7.03 (1H, d, J=8Hz), 7.42 (1H, m, J=8Hz), 7.56 (2H, m), 8.18 (1H, d, J=8Hz), 8.70 (1H, d, J=8Hz); ¹³C NMR, (75MHz, CDCl ₃) δ 111.69,112.56,120.85 (Cq), 121.56 (Cq), 122.57,124.00,126.72,129.83,136.30 (Cq), 143.98 (Cq), 146.79 (Cq), 156.99 (Cq); MS (EI) m/z 229.1M ⁺HRMS C ₁₂H ₇NO ₄[M+H] ⁺, calculated value: 229.0370, measured value: 229.0369.

(e) three fluoro-methylsulfonic acid 1-nitro-diphenylene-oxide-4-base ester (11)

With 1-nitro-diphenylene-oxide-4-alcohol (3,01g; 13.36mmol) be dissolved among the DCM (50ml), be cooled to-40 ℃, and the adding triethylamine (5.5ml, 40mmol).After 5 minutes, (3.45ml 20mmol) dropwise joins in this reaction mixture with trifluoromethanesulfanhydride anhydride.In adition process, the temperature of this reaction mixture is remained on below-30 ℃.After 3 hours, this reaction mixture with saturated sodium carbonate solution (50ml) washing, is used DCM (3 * 30ml) extractions.By dried over mgso, evaporation obtains brown solid with organic layer.This solid is carried out purifying through silicon-dioxide plug, use DCM/EP (6: 4) as eluent, obtain the title compound (4.536g, 12.56mmol, 94%) of white solid state: R _f=0.33 (DCM-EP 1: 4); Mp:102-103 ℃; λ _Max(EtOH)/nm 243; IR (cm ^-1) 1643,1572,1528,1488,1427,1348,1317,1246,1209,1132,1068,981,921,828,792,742,700; ¹H NMR, (300MHz, CDCl ₃) δ 7.39 (1H, m); 7.55 (1H, d, J=8Hz); 7.65 (2H, m); 8.20 (1H, d, J=8Hz); 8.51 (1H, d, J=8Hz); ¹³C NMR, (75MHz, CDCl ₃) δ 112.35,117.12 (CF ₃), 119.49,120.55 (CF ₃), 120.75,121.38 (CF ₃), 122.81 (CF ₃), 124.92,126.60,131.24,137.88 (Cq), 142.32 (Cq), 148.21 (Cq), 157.99 (Cq); MS (EI) m/z 361.1M ⁺HRMS C ₁₃H ₆F ₃NO ₆S[M+H] ⁺, calculated value: 360.9862, measured value: 360.9861.

(f) 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls) diphenylene-oxide (12)

In the Schlenk pipe, will two (tetramethyl ethylene ketone) two boron (3.075mg, 12.11mmol) and potassium acetate (1.17g 18.16mmol) is dissolved in the diox (10ml) and outgases.Simultaneously, with 4-hydroxyl-1-nitro-diphenylene-oxide 4-O-triflate (2.186g, 6.05mmol), PdCl ₂(247mg, 0.30mmol) (167mg 0.30mmol) is dissolved in the diox (10ml) and the degassing two (triphen phosphino-) ferrocene with two (triphen phosphino-) ferrocene.In the Schlenk pipe, two kinds of solution are mixed, stir, in 110 ℃ of heating 18 hours.After the cooling, add DCM (20ml).With solution with water (20ml) washing, pass through dried over mgso then, evaporation.Resistates is carried out purifying as eluent and realization to the gradient of DCM/AcOH (98: 2) by flash chromatography, use DCM/EP (6: 4), pass whole products of post with recovery.This product is concentrated, and (3 * 20ml) washings are to remove acetate with yellow soda ash.After the evaporation, recrystallization in DCM obtains yellow crystalline product (1.783mg, 5.26mmol, 87%): R _f=0.30 (AcOEt-EP 1: 49); Mp:185 ℃; λ _Max(EtOH)/nm 347; IR (cm ^-1) 2981,1701,1624,1597,1521,1474,1447,1369,1329,1309,1192,1172,1137,1043,979,883,852,818,785,754,732,663; ¹H NMR, (300MHz, CDCl ₃) δ 1.48 (12H, s, 4 * CH ₃), 7.46 (1H, t, J=6Hz), 7.63 (1H, t, J=6Hz), 7.76 (1H, d, J=9Hz), 8.01 (1H, d, J=9Hz), 8.19 (1H, d, J=9Hz), 8.66 (1H, d, J=9Hz); ¹³C NMR, (125MHz, CDCl ₃) δ 24.82 (4 * CH ₃), 84.78 (2 * Cq-O), 112.14,118.07 (Cq), 118.61,120.29 (Cq), 123.42,125.82,129.47,133.50,144.81 (Cq), 157.33 (Cq), 161.39 (Cq); MS (EI) m/z 339.2 M ⁺HRMS C ₁₈H ₁₈BNO ₅[M+H] ⁺, calculated value: 339.1273, measured value: 339.1270.

(iii) 9-trifluoromethanesulfonic acid ester group-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (17) is synthetic

(a) 2,9-dihydroxyl-pyrido [2,1-a] pyrimidin-4-ones (14)

To be dissolved in propanedioic acid in the bromobenzene (37ml) two-(2,4,6-three chloro-phenyl) ester (17.33g; 37.5mmol) and 3-hydroxyl-2-amino-pyridine (13) (4.12g; 37.5mmol) mixture heating up refluxed 3 hours.After the cooling, this reaction mixture is filtered, the solid washing with alcohol.This solid is dissolved among the 1M NaOH, adds several AcOH, be settled out faint yellow solid-state product (6.53g).Productive rate=98%.m.p.:320 ℃ (degraded); R _f=0.11, MeOH: DCM (3: 17); UV: λ _Max=252nm; IR:(cm ^-1) 2862,1688,1564,1374,1295,1102,783; ¹H NMR, (DMSO, 300MHz), δ (ppm): 5.22 (1H, s, CH-3), 7.12 (1H, t, J _H6-H7=7Hz, H _Aromatics-7), 7.27 (1H, d, J _H7-H8=8Hz, H _Aromatics-8), 8.43 (1H, d, J _H6-H7=7Hz, H _Aromatics-6); ¹³C NMR, (CDCl ₃, 75MHz), δ (ppm): 103.25,116.46,117.05,119.03,143.97,148.82,157.26,157.50.

(b) 2-chloro-9-hydroxyl-pyrido [2,1-a] pyrimidin-4-ones (15)

In round-bottomed flask, with 2,9-dihydroxyl-pyrido [2,1-a] pyrimidin-4-ones (14) (1.07g; 6.0mmol) be dissolved in the phosphoryl chloride (7.5ml).This solution is heated to backflow reaches 48 hours.After the cooling, this reaction mixture is inclined carefully to icy water (100ml), transfer to pH7 by adding saturated sodium carbonate solution.With the water layer dichloromethane extraction.By dried over mgso, evaporation obtains brown solid with organic layer.This solid is carried out purifying by flash chromatography, use methylene dichloride as eluent, obtain the title compound (712mg) of white solid state.Productive rate=60%.m.p.:162 ℃; R _f=0.34, MeOH: DCM (1: 19); Mass spectrum: (m/z) 196.93[M+1] ⁺(Rt=4.67 minute, 12 minutes gradients); UV: λ _Max=210nm; IR:(cm ^-1) 3103,1684,1630,1511,1458,1297,1105; ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 6.4 (1H, s, CH-3), 7.11 (1H, t, J _H6-H7=7Hz, H _Aromatics-7), 7.25 (1H, d, J _H7-H8=8Hz, H _Aromatics-8), 8.51 (1H, d, J _H6-H7=7Hz, H _Aromatics-6); ¹³C NMR, (CDCl ₃, 75MHz), δ (ppm): 103.25,116.46,117.05,119.03,143.97,148.82,157.26,157.50.

(c) 9-hydroxyl-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (16)

In round-bottomed flask, with 2-chloro-9-hydroxyl-pyrido [2,1-a] pyrimidin-4-ones (15) (141.7mg; 0.721mmol) and morpholine (314 μ L; 3.605mmol) be dissolved in the ethanol (5ml).Under vigorous stirring, this solution is heated to backflow and reaches 18 hours.After the cooling, evaporating solvent.With the thick solid of yellow recrystallization in ethanol, obtain the 173.8mg white crystals.Productive rate=97%.m.p.:245 ℃; R _f=0.27, MeOH: DCM (1: 19); Mass spectrum: (m/z) 248.08[M+1] ⁺(Rt=4.92 minute, 12 minutes gradients); UV: λ _Max=267nm; IR:(cm ^-1) 3302,1690,1644,1551,1427,1224,1110.; ¹H NMR, (CDCl ₃, 500MHz), δ (ppm): 3.56 (4H, m, N-CH ₂-morpholine), 3.75 (4H, m, O-CH ₂-morpholine), 5.55 (1H, s, CH-3), 6.80 (1H, t, J _H6-H7=7Hz, H _Aromatics-7), 7.02 (1H, dd, J _H7-H8=8Hz, J _H6-H8=1.3Hz, H _Aromatics-8), 7.33 (1H, s, OH), 8.37 (1H, dd, J _H6-H7=7Hz, J _H6-H8=1.3Hz, H _Aromatics-6); ¹³C NMR, (CDCl ₃, 125MHz), δ (ppm): 45.27,67.02,82.16,113.46,114.18,119.05,143.00,147.51,159.00,161.00.

(d) 9-trifluoromethanesulfonic acid ester group-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (17)

In having three neck round-bottomed flasks of thermometer, with 9-hydroxyl-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (16) (2.11g; 8.54mmol) be dissolved among the DCM (70mL), be cooled to-30 ℃, add triethylamine (3.572ml; 25.63mmol).After 5 minutes, go through to be dissolved in trifluoromethanesulfanhydride anhydride (2.101ml among the 10ml DCM by addition funnel in 30 minutes; 12.81mmol) dropwise join in this reaction mixture.In adition process, the temperature of this reaction mixture is remained on below-20 ℃.After 3 hours, with the saturated Na of this reaction mixture ₂CO ₃Solution (50ml) washing is with DCM (3 * 30ml) extractions.By dried over mgso, evaporation obtains brown solid with organic layer.This solid is carried out purifying by flash chromatography, use methylene dichloride as eluent, obtain orange solid-state title compound (2.91g).Productive rate=90%.m.p.:146-147 ℃; R _f=0.42; MeOH: DCM (1: 19); Mass spectrum: (m/z) 380.16[M+1] ⁺(Rt=3.34 minute, 12 minutes gradients); UV: λ _Max=271nm; IR:(cm ^-1) 1705,1644,1551,1189,1112,939,769; ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 3.56 (4H, m, N-CH ₂-morpholine), 3.71 (4H, m, O-CH ₂-morpholine), 5.53 (1H, s, CH-3), 6.80 (1H, t, J _H6-H7=7Hz, H _Aromatics-7), 7.46 (1H, dd, J _H7-H8=8Hz, J _H6-H8=1.3Hz, H _Aromatics-8), 8.79 (1H, dd, J _H6-H7=7Hz, J _H6-H8=1.3Hz, H _Aromatics-6); ¹³CNMR, (CDCl ₃, 75MHz), δ (ppm): 45.19,66.87,81.76,110.16,112.61,116.85,121.10,125.34,127.86,128.13,141.46,145.79,158.07,160.42.

(iv) 8-bromo-2-morpholine-4-base-1H-quinoline-4-ketone (21) is synthetic

(a) 5-(two-methyl sulfane base-methylene radical)-2,2-dimethyl-[1,3] diox-4,6-diketone (18)

In the 250ml two-neck bottle, form well-beaten 2,2-dimethyl-1,3-diox-4,6-diketone (12) (Meldrum acid) (4.09g; 28.4mmol) solution in DMSO (14ml).With triethylamine (7.92ml; 56.8mmol) and dithiocarbonic anhydride (1.71ml; 28.4mmol) join continuously in this mixture fast.Then in room temperature with this mixture vigorous stirring 1 hour, in ice bath, cool off then.Cooling off under (ice bath) methyl iodide (3.54ml; 56.8ml) slowly join in this reaction mixture.When adding is finished, make this reaction mixture be warmed to room temperature, stirred in addition 4 hours, use icy water (25ml) dilution then.Scrape this mixture, make the product precipitation, this product is filtered, wash with gasoline.Obtain yellow solid-state product (2.76g), it is enough pure, can be used for subsequent reaction.Productive rate=45%.m.p.:118 ℃ of (document ²⁸: 116-118 ℃); IR:(cm ^-1) 3728,1668,1373,1302,1264,1199; ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 1.54 (6H, s, 2CH ₃), 2.58 (6H, s, 2CH ₃-S); ¹³C NMR, (CDCl ₃, 75MHz), δ (ppm): 21.86,27.22,103.32,160.33,194.

(b) 5-[2-bromobenzene amido-(methylthio group)-methylene radical]-2,2-dimethyl-4,6-diketone (19)

In the 10ml round-bottomed flask that has water cooler and nitrogen bubble device, with two methylthio group subunit isopropylidene malonates (18) (900mg; 3.63mmol) and 2-bromaniline (15) (624mg; 3.63mmol) be dissolved in the 2,2,2 tfifluoroethyl alcohol (3.6ml).This mixture is stirred, and being heated to refluxes reaches 22 hours.After the cooling, evaporating solvent.With resistates recrystallization from methyl alcohol, obtain white crystalline title compound (1.192g).Productive rate=88%.m.p.:159 ℃; R _f=0.31, DCM; IR:(cm ^-1) 2990,1706,1653,1535,1370,1199; UV: λ _Max=313nm; ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 1.69 (6H, s, 2CH ₃), 2.15 (3H, s, CH ₃-S), 7.18 (1H, dt, J _H4-H5=8Hz, J _H4-H6=2Hz, H _Aromatics-4), 7.35 (2H, m, H _Aromatics-5 and H _Aromatics-6), 7,61 (1H, dd, J _H3-H4=8Hz, J _H3-H5=1.2Hz, H _Aromatics-3), 12.51 (1H, s, N-H); ¹³C NMR, (CDCl ₃, 75MHz), δ (ppm): 18.75,26.48,87.54,103.32,120.45,127.78,128.46,129.48,133.57,136.91,163.87,178.70.

(c) 5-[(2-bromo-anilino)-and morpholine-4-base-methylene radical]-2,2-dimethyl-[1,3] diox-4,6-diketone (20)

In the 10ml round-bottomed flask that has water cooler and nitrogen bubble device, with 5-[2-bromobenzene amido-(methylthio group)-methylene radical]-2,2-dimethyl-4,6-diketone (19) (234mg; 0.629mmol) and morpholine (110 μ L; 1.257mmol) be dissolved in the 2,2,2 tfifluoroethyl alcohol (1ml).This mixture is stirred, and being heated to refluxes reaches 18 hours.After the cooling, evaporating solvent.With resistates recrystallization from methyl alcohol, obtain white crystalline title compound (0.124g).Productive rate=50%.m.p.:212-213 ℃; R _f=0.05; DCM; IR:(cm ^-1) 1627,1342,1305,1100,1022,934; UV: λ _Max=241nm; ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 1.77 (6H, s, 2CH ₃), 3.24 (4H, t, J _Ab=5Hz, 2CH ₂-N morpholine), 3.66 (4H, t, J _Ab=5Hz, 2CH ₂-O morpholine), 7.18 (2H, m, H _Aromatics-4 and H _{Virtue Family}-6), 7.40 (1H, t, J _H5-H6=8Hz, H _Aromatics-5), 7.69 (1H, dd, J _H3-H4=8Hz, J _H3-H5=1.4Hz, H _Aromatics-3), 9.62 (1H, s, N-H); ¹³C NMR, (CDCl ₃, 75MHz), δ (ppm): 26.83,51.14,65.62,87.54,102.84,120.45,127.15,128.92,129.03,134.48,138.46,164.92,178.70.

(d) 8-bromo-2-morpholine-4-base-1H-quinoline-4-ketone (21)

In the Schlenk pipe, with 5-[(2-bromo-anilino)-morpholine-4-base-methylene radical]-2,2-dimethyl-[1,3] diox-4,6-diketone (20) (103.3mg; 0.2513mmol) be dissolved in the phenyl ether (0.7ml).This mixture is stirred, and being heated to refluxes reaches 4 hours.After the cooling, add sherwood oil.By the collected at suction product.Resistates is carried out purifying by flash chromatography, use methylene chloride (95: 5) as eluent.Obtain brown buttery product (65.1mg).Productive rate=84%.R _f=0.25, MeOH: DCM (1: 19); Mass spectrum: (m/z) 310.98[M+1] ⁺(Rt=5.24 minute, 12 minutes gradients); IR:(cm ^-1) 3395,2959,2849,1617,1577,1487,1421,1384,1327,1263,1229,1188,1152,1111,1066,999,902,785; UV: λ _Max=254nm; ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 3.72 (4H, t, J _Ab=5Hz, 2CH ₂-N morpholine), 3.75 (4H, t, J _Ab=5Hz, 2CH ₂-O morpholine), 5.95 (1H, s, H-3), 7.04 (1H, t, J _H6-H7=8Hz, H _Aromatics-6), 7.69 (1H, dd, J _H6-H7=8Hz, J _H5-H7=1.3Hz, H _Aromatics-7), 8.09 (1H, d, J _H5-H6=8Hz, H _Aromatics-5); ¹³CNMR, (CDCl ₃, 75MHz), δ (ppm): 46.35,66.59,92.50,114.53,123,123.50,124.73,134.45,138,156.06,172.6.

(v) 9-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (23) is synthetic

(a) 9-(1-nitro-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (22)

In the Schlenk pipe, with 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls) dibenzothiophene (7) (983mg; 2.8mmol) and cesium carbonate (2.705g; 8.3mmol) be dissolved among the THF (8ml).Argon gas is blasted in this solution, with this solution supersound process 15 minutes.Simultaneously, with 9-trifluoromethanesulfonic acid ester group-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (6) (1.154g; 3.045mmol) and PdCl ₂Two (triphen phosphino-) ferrocene (112.7mg; 0.138mmol) be dissolved among the THF (8ml).Argon gas is blasted in this solution, with this solution supersound process 15 minutes.In the Schlenk pipe, two kinds of solution are mixed, stir, in 80 ℃ of heating 18 hours.After the cooling, add DCM (20ml).With solution with water (20ml) washing, pass through dried over mgso then, evaporation.Resistates is carried out purifying by flash chromatography, use ethyl acetate/DCM (1: 1) as eluent.After the evaporation, obtain yellow solid-state product (1.168g).Productive rate=92%.R _f=0.37; AcOEt: DCM (1: 1); Mass spectrum: (m/z) 459.3[M+1] ⁺(Rt=4.69 minute).

(b) 9-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (23)

In round-bottomed flask, with 9-(1-nitro-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (17) (618.8mg; 1.351mmol) be suspended among the AcOH (10ml).With zinc powder (883.3mg; 13.51mmol) join in this solution, in room temperature this reactant is stirred and spend the night.Pass through Celite ^TMThis reaction mixture is filtered, with methyl alcohol (4 * 50ml) and DCM (4 * 50ml) continuous washing.The organic layer that vapourisation under reduced pressure merges.Resistates water (100ml) is stirred, and (25ml) joins in this solution with ammoniacal liquor.Collect the gained throw out by filtering.With the resistates drying, this resistates need not to be further purified.Obtain yellow solid-state product (575.3mg).Productive rate=99%.R _f=0.36; AcOEt: DCM (1: 1); Mass spectrum: (m/z) 429.47[M+1] ⁺(Rt=4.17 minute).

(vi) 8-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-1H-quinoline-4-ketone (25) is synthetic

(a) 2-morpholine-4-base-8-(1-nitro-dibenzothiophene-4-yl)-1H-quinoline-4-ketone (24)

In the Schlenk pipe, with 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls) dibenzothiophene (6) (983mg; 2.768mmol) and cesium carbonate (2.705g; 8.3047mmol) be dissolved among the THF (8ml).Argon gas is blasted in this solution, with this solution supersound process 15 minutes.Simultaneously, with 8-bromo-2-morpholine-4-base-1H-quinoline-4-ketone (21) (941.4mg; 3.045mmol) and PdCl ₂Two (triphen phosphino-) ferrocene (112.7mg; 0.138mmol) be dissolved among the THF (8ml).Argon gas is blasted in this solution, with this solution supersound process 15 minutes.In the Schlenk pipe, two kinds of solution are mixed, stir, in 80 ℃ of heating 18 hours.After the cooling, add DCM (20ml).With solution with water (20ml) washing, pass through dried over mgso then, evaporation.Resistates is carried out purifying by flash chromatography, use ethyl acetate/DCM (1: 1) as eluent.After the evaporation, obtain yellow solid-state product (255.5mg).Productive rate=20%.R _f=0.24, AcOEt: DCM (1: 1); Mass spectrum: (m/z) 4.58.4[M+1] ⁺(Rt=5.33 minute).

(b) 8-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-1H-quinoline-4-ketone (25)

In round-bottomed flask, with 2-morpholine-4-base-8-(1-nitro-dibenzothiophene-4-yl)-1H-quinoline-4-ketone (24) (365mg; 0.798mmol) be suspended among the AcOH (5ml).With zinc powder (5223.3mg; 7.98mmol) join in this solution, in room temperature this reactant is stirred and spend the night.Pass through Celite ^TMThis reaction mixture is filtered, with methyl alcohol (4 * 25ml) and DCM (4 * 25ml) continuous washing.The organic layer that vapourisation under reduced pressure merges.Resistates water (50ml) is stirred, and (15ml) joins in this solution with ammoniacal liquor.Collect the gained throw out by filtering.With the resistates drying, this resistates need not to be further purified.Obtain yellow solid-state product (291.3mg).Productive rate=85.4%.Mass spectrum: (m/z) 428.4[M+1] ⁺(Rt=3.83 minute).

(vii) 9-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (27) is synthetic

(a) 9-(1-nitro-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (26)

In the Schlenk pipe, with 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls) diphenylene-oxide (12) (500mg, 1.47mmol) and salt of wormwood (480mg 3.69mmol) is dissolved in the diox (10ml) and the degassing.Simultaneously, with 9-hydroxyl-2-morpholine-4-base-pyrido [1,2-a] pyrimidin-4-one 9-O-triflate (17) (466mg, 1.23mmol) and Pd (PPh ₃) ₄(71mg 0.06mmol) is dissolved in the diox (10ml) and the degassing.In microwave container, two kinds of solution are mixed, this microwave container was placed microwave reactor 30 minutes in 180 ℃.After the cooling, add DCM (20ml).With solution with water (20ml) washing, pass through dried over mgso then, evaporation.Resistates is ground in hot methanol, filter, obtain the solid-state product of brown (375mg, 0.85mmol, 69%): R _f=0.51 (AcOEt); Mp:262 ℃; λ _Max(EtOH)/nm 268; IR (cm ^-1) 1706,1673,1631,1599,1541,1511,1430,1338,1306,1230,1194,1116,1070,1028,999,975,860; ¹H NMR, (300MHz, CDCl ₃) δ 3.26 (4H, m, 2 * N-CH ₂-morpholine), 3.47 (4H, m, 2 * O-CH ₂-morpholine), 5.57 (1H, s, H-3), 7.03 (1H, t, J _H6-H7=7Hz, H-7), 7.44 (1H, t, J=7Hz), 7.58 (2H, m), 7.74 (1H, d, J=7Hz), 7.86 (1H, d, J=7Hz), 8.24 (1H, d, J=8Hz), 8.66 (1H, d, J=8Hz), 9.10 (1H, d, J=8Hz, H-6); ¹³CNMR, (125MHz, CDCl ₃) δ 44.41 (2 * CH ₂-N-morpholine), 66.32 (2 * CH ₂-O-morpholine), 81.09 (C-3), 111.58,112.05,119.26,120.67,123.89,126.28,127.57,128.32,128.76,128.95,130.01,138.39,142.88,143.92,148.48,154.66,157.04 (Cq), 158.67 (Cq), 160.28 (Cq); MS (EI) m/z 443.35M ⁺HRMS C ₂₄H ₁₉N ₄O ₅[M+H] ⁺, calculated value: 443.1350, measured value: 443.1352.

(b) 9-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (27)

In round-bottomed flask, (300mg 0.68mmol) is suspended among the AcOH (5ml) with 9-(1-nitro-diphenylene-oxide-4-yl)-2-morpholine-4-base-pyrido [1,2-a] pyrimidin-4-one (26).(445mg 6.8mmol) joins in this solution, in room temperature this reactant is stirred and spends the night with zinc powder.By diatomite this reaction mixture is filtered, with methyl alcohol (4 * 50ml) and DCM (4 * 50ml) continuous washing.The organic layer that vapourisation under reduced pressure merges.Resistates water (100ml) is stirred, and (25ml) joins in this solution with ammoniacal liquor.Collect the gained throw out by filtering.This solid is carried out purifying by flash chromatography, use AcOEt as eluent, obtain the title compound (269mg, 0.65mmol, 96%) of white solid state: R _f=0.32 (AcOEt); Mp:294 ℃; λ _Max(EtOH)/nm 238; IR (cm ^-1) 3340,3224,2937,2872,2258,1697,1637,1623,1543,1493,1440,1373,1309,1258,1234,1191,1150,1109,1073,999,909,856,776; ¹H NMR, (300MHz, MeOD) δ 3.36 (4H, m, 2 * N-CH ₂-morpholine), 3.52 (4H, m, 2 * O-CH ₂-morpholine), 6.72 (1H, d, J=7Hz), 6.88 (1H, t, J=7Hz), 7.31-7.47 (5H, m), 7.90 (2H, m), 8.90 (1H, d, J=7Hz); ¹³C NMR, (75MHz, MeOD) δ 46.23 (2 * CH ₂-N-morpholine), 68.02 (2 * CH ₂-O-morpholine), 110.44,112.78,114.57,122.46,124.53,125.58 (Cq), 127.53,128.01,131.81,133.58,138.88,145.06 (Cq), 150.82 (Cq), 156.70 (Cq), 156.90 (Cq), 161.68 (Cq), 162.11 (Cq); MS (EI) m/z 413.19M ⁺0.86mol C ₂₄H ₂₀N ₄O ₃+ 0.14mol MeOH, ultimate analysis calculated value: C, 69.48, H, 4.99, N, 13.41. measured value: C, 69.22, H, 4.79, N, 13.38; HRMS C ₂₄H ₂₁N ₄O ₃[M+H] ⁺, calculated value: 413.1608, measured value: 413.1609.

(viii) 9-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (29) is synthetic

(a) 2-morpholine-4-base-8-(1-nitro-diphenylene-oxide-4-yl)-1H-quinoline-4-ketone (28)

In the Schlenk pipe, with 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls) diphenylene-oxide (12) (500mg, 1.47mmol) and salt of wormwood (480mg 3.69mmol) is dissolved in the diox (10ml) and the degassing.Simultaneously, with 8-bromo-2-morpholine-4-base-1H-quinoline-4-ketone (21) (380mg, 1.23mmol) and Pd (PPh ₃) ₄(71mg 0.06mmol) is dissolved in the diox (10ml) and the degassing.In microwave container, two kinds of solution are mixed, this microwave container was placed microwave reactor 30 minutes in 180 ℃.After the cooling, add DCM (20ml).With solution with water (20ml) washing, pass through dried over mgso then, evaporation.Resistates is carried out purifying by flash chromatography, use AcOEt/EP (8: 2) as eluent, obtain yellow solid-state title compound (303mg, 0.74mmol, 60%): R _f=0.67 (AcOEt); Mp:247 ℃; λ _Max(EtOH)/nm 251; IR (cm ^-1) 3421,2852,2360,2333,1614,1573,1500,1435,1429,1348,1309,1233,1199,1152,1124,1039,991,917,866,823; ¹H NMR, (300MHz, CDCl ₃) δ 3.04 (4H, m, 2 * CH ₂-N-morpholine), 3.54 (4H, m, 2 * CH ₂-O-morpholine), 5.92 (1H, s, H-3), 7.19-7.45 (2H, m), 7.51-7.58 (2H, m), 7.65-7.70 (2H, m), 8.31 (2H, m), 8.67 (1H, d, J=7Hz); MS (EI) m/z 442.29 M ⁺HRMS C ₂₅H ₂₀N ₃O ₅[M+H] ⁺, calculated value: 442.1397, measured value: 442.1398.

(b) 8-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-1H-quinoline-4-ketone (29)

In round-bottomed flask, (290mg 0.66mmol) is suspended in the acetate (5ml) with 2-morpholine-4-base-8-(1-nitro-dibenzothiophene-4-yl)-1H-quinoline-4-ketone (28).(430mg 6.58mmol) joins in this solution, in room temperature this reactant is stirred and spends the night with zinc powder.By diatomite this reaction mixture is filtered, use methyl alcohol (4 * 25ml) and DCM (4 * 25ml) continuous washing then.The organic layer that vapourisation under reduced pressure merges.Resistates water (50ml) is stirred, and (15ml) joins in this solution with ammoniacal liquor.Collect the gained throw out by filtering.With the resistates drying, this resistates need not to be further purified.Obtain brown buttery product (246mg, 0.60mmol, 91%): R _f=0.44 (AcOEt); λ _Max(EtOH)/nm 315; IR (cm ^-1) 1708,1572,1374,1278,1190,1116,1049,1010,931,880,827,748,722,688,688; ¹H NMR, (300MHz, CDCl ₃) δ 3.11 (4H, m, 2 * CH ₂-N-morpholine), 3.58 (4H, m, 2 * CH ₂-O-morpholine), 5.98 (1H, s, H-3), 6.85 (1H, d, J=8Hz), 7.25-7.51 (3H, m), 7.52-7.68 (3H, m), 8.05 (1H, t, J=8Hz), 8.67 (1H, d, J=8Hz); ¹³C NMR, (75MHz, MeOD) δ 48.02 (2 * CH ₂-N-morpholine), 67.44 (2 * CH ₂-O-morpholine), 111.94,112.63,122.96,125.05,125.57,127.63,127.87,131.55,132.68,135.32,146.21,156.79; MS (EI) m/z 412.25 M ⁺HRMSC ₂₅H ₂₂N ₃O ₃[M+H] ⁺, calculated value: 412.1656, measured value: 412.1654.

(ix) 9-(1-hydroxyl-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (31) is synthetic

In round-bottomed flask, with 9-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (23) (137.2mg; 0.32mmol) be suspended in the ethanol (15ml).Dropwise add HBF in room temperature ₄(664 μ L; 4.81mmol).Stir after 15 minutes, this reaction mixture becomes settled solution, in 0 ℃ this solution is cooled off, and adds nitrite tert-butyl (76.1 μ L; 0.64mmol).After 30 minutes, reaction mixture is diluted with ether (25ml).There is throw out to form,, uses ether (2 * 5ml) washings, drying then its filtration.This solid joined contain Red copper oxide (45.78mg; 0.32mmol) cupric nitrate (23.193g; 96mmol) in the solution in water (500ml), in stirring at room 1 hour.The aqueous solution is filtered, obtain brown solid.Resistates is carried out purifying by flash chromatography, use methyl alcohol/DCM (2: 98) as eluent.The by product (22.5mg) that separation is produced by deamination, productive rate is 17%.Obtain yellow solid-state product (26.3mg).Productive rate=19%.Mass spectrum: (m/z) 430.3[M+1] ⁺(Rt=4.23 minute).

(x) 8-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-1-chromene-4-ketone (33) is synthetic

(a) 2-morpholine-4-base-8-(1-nitro-diphenylene-oxide-4-yl)-1-chromene-4-ketone (32)

To 1-nitro-4-(4,4,5,5-tetramethyl--1,3,2-two oxa-boron heterocycle pentane-2-yls)-(12,0.30mmol 0.10g) adds three fluoro-methylsulfonic acid 2-morpholine-4-base-4-oxo-4H-1-chromenes-8-base ester (0.24mmol to diphenylene-oxide in the solution in no water diox (2ml), 0.091g, see WO03/024949), salt of wormwood (0.6mmol, 0.083g) and four (triphenyl phosphine) palladium (0.015mmol, 0.018g).With the reaction vessel sealing, in the influence heating down of microwave radiation (140 ℃, 10 minutes, the low absorption set).After finishing, reaction mixture is filtered by the thin pad of silicon-dioxide, filter cake CH ₂Cl ₂Washing.Solvent removed in vacuo obtains filbert solid-state resistates (0.13g, 100%) then, and it uses without being further purified.M/z 443.4[M+H] ⁺(Rt=4.76 minute).

(b) 8-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-1-chromene-4-ketone (33)

Go through 10 fens clockwise 2-morpholine-4-base-8-(1-nitro-diphenylene-oxide-4-yl)-1-chromene-4-ketone (32,2.5mmol, 1.11g) add in the solution in acetate (50ml) in batches zinc powder (25mmol, 1.64g).In room temperature this mixture was stirred 2 hours then, subsequently it is passed through Celite ^TMPad filters, with methyl alcohol (20ml) and CH ₂Cl ₂(20ml) washing.Solvent removed in vacuo obtains soup compound, then it is diluted with ammonium hydroxide (30ml), by removing by filter the gained solid.Resistates is passed through flash chromatography (SiO ₂), use MeOH: CH ₂Cl ₂As eluent carry out purifying at-1: 99, obtains the required product (75%) of analytical pure form.M/z 413.5[M+H] ⁺(Rt=4.27 minute).

Embodiment 1: synthesized by the parallel of 9-trifluoromethanesulfonic acid ester group-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (17)

With suitable for boric acid (boronic acid) (0.395mmol) and salt of wormwood (109.3mg; 0.7914mmol) join in the rotating disc type pipe (carousel tube).With the bottle emptying, clean with argon gas.This operation is carried out 3 times.In the Schlenk pipe, with 9-trifluoromethanesulfonic acid ester group-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (17) (100mg; 0.2638mmol; Each rotating disc type pipe) is dissolved in diox (2ml; Each rotating disc type pipe) in.Argon gas is blasted in this solution, with this solution supersound process 15 minutes.In another Schlenk pipe, with four-(triphenyl phosphine)-palladium (15.2mg; 0.013mmol; Each rotating disc type pipe) is dissolved in diox (2ml; Each rotating disc type pipe) in.Argon gas is blasted in this solution, with this solution supersound process 15 minutes.In the rotating disc type pipe, each solution of 2ml is mixed, in 95 ℃ of stirring heating 48 hours.After the cooling, solution is filtered by the Radleys Discovery Technologie solid-phase extraction column that places the 500mg silicon-dioxide in the stalker parallel purification system.Post with ethyl acetate (20ml) washing, is collected as phase 1.Then post (20ml) is washed with methylene chloride (85: 15), collect as phase 2.By LC/MS the biphase product is detected.In some cases, 2 only contain impurity mutually, in other cases, two-phase are merged evaporation.Depend on product, carry out purifying by HPLC or by flash chromatography.

NMR result

34a: ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 3.36 (4H, m, N-CH ₂-morpholine); 3.56 (4H, m, O-CH ₂-morpholine); 5.64 (1H, s, CH-3); 7.01 (1H, t, J _H6-H7=7Hz, H _{Virtue Family}-7); 7.45-7.49 (2H, m); 7.56-7.57 (2H, m); 7.79-7.82 (1H, m); 7.86-7.89 (1H, dd); 8.19-8.23 (2H, m); 9.04 (1H, dd).

¹³C-NMR, (CDCl ₃, 300MHz), δ (ppm): 44.83 (CH ₂-N-morpholine); (66.84 CH-O-morpholine); 81.37 (CH-3); 112.47 (CH-7); 121.69; 122.10; 122.92; 124.62; 124.85; 127.39; 128.27; 128.91; 132.41; 134.77; 135.88; 136.36; 137.54; 139.60; 140.13; 148.84; 159.38 (Cq); 160.54 (C=O).

34b: ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 3.35 (4H, t, N-CH ₂-morpholine); 3.54 (4H, t, O-CH ₂-morpholine); 5.66 (1H, s, CH-3); 7.08 (1H, t, J _H6-H7=7Hz, H _Aromatics-7); 7.38-7.51 (4H, m, H _Aromatics-diphenylene-oxide); 7.67 (1H, dd, J _H7-H8=7.6Hz, J _H6-H8=1.3Hz, H _Aromatics-8); 7.94 (1H, dd, J=7Hz, J=1.6Hz, H _Aromatics-diphenylene-oxide); 8.01-8.05 (2H, m, H _Aromatics-diphenylene-oxide); 9.06 (1H, dd, J _H6-H7=7Hz, J _H6-H8=1.3Hz, H _Aromatics-6).

¹³C-NMR, (CDCl ₃, 300MHz), δ (ppm): 44.80 (CH ₂-N-morpholine); (66.80 CH-O-morpholine); 81.37 (CH-3); 111.98; 112.65,121.14,121.75,122.76; 123.31; 124.38; 124.89; 127.80; 128.03; 129.28; 131.42; 138.06; 156.30; 159.48; 160.69.

21c: ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 3.42 (4H, m, N-CH ₂-morpholine); 3.56 (4H, m, O-CH ₂-morpholine); 5.40 (1H, s, CH-3); 6.87 (1H, t, J _H6-H7=7Hz, H _{Virtue Family}-7); 7.26-7.83 (10H, m, H _Aromatics-xenyl and H _Aromatics-8); 8.87 (1H, dd, J _H6-H7=7.1Hz, J _H6-H8=1.6Hz, H _Aromatics-6).

Embodiment 2: synthesized by the parallel of 8-bromo-2-morpholine-4-base-1H-quinoline-4-ketone (21)

With suitable for boric acid (0.486mmol) and salt of wormwood (269.2mg; 1.946mmol) join in the rotating disc type pipe.With the bottle emptying, clean with argon gas.This operation is carried out 3 times.In the Schlenk pipe, with 8-bromo-2-morpholine-4-base-1H-quinoline-4-ketone (21) (100mg; 0.324mmol; Each rotating disc type pipe) is dissolved in diox (2ml; Each rotating disc type pipe) in.Argon gas is blasted in this solution, with this solution supersound process 15 minutes.In another Schlenk pipe, with four-(triphenyl phosphine)-palladium (18.7mg; 0.016mmol; Each rotating disc type pipe) is dissolved in diox (2ml; Each rotating disc type pipe) in.Argon gas is blasted in this solution, with this solution supersound process 15 minutes.In the rotating disc type pipe, each solution of 2ml is mixed, in 95 ℃ of stirring heating 48 hours.After the cooling, solution is filtered by the Radleys Discovery Technologie solid-phase extraction column that places the 500mg silicon-dioxide in the stalker parallel purification system.Post with ethyl acetate (20ml) washing, is collected as phase 1.Then post (20ml) is washed with methylene chloride (85: 15), collect as phase 2.By LC/MS the biphase product is detected.In some cases, 2 only contain impurity mutually, in other cases, two-phase are merged evaporation.Depend on product, carry out purifying by HPLC or by flash chromatography.

NMR result

35a: ¹H-NMR, (CDCl ₃.300MHz), δ (ppm): 2.93 (4H, t, J=5Hz, CH ₂-N morpholine), 3.57 (4H, t, J=5Hz, CH ₂-O morpholine), 5.67 (1H, s, H-3), 7.34 (t, 1H, J=8Hz, H _Aromatics), 7.42-7.74 (m, 8H, H _Aromatics), 8.15-8.23 (m, 2H, H _Aromatics), 8.33 (d, 1H, J=8Hz, H _Aromatics).

¹³C-NMR, (CDCl ₃, 300MHz), δ (ppm): 46.94 (CH ₂-N morpholine), 66.22 (CH ₂-O morpholine), 93.44 (CH-3), 122.38 (CH), 122.46 (CH), 123.30 (CH), 123.43 (CH), 124.62 (CH), 125.29 (CH), 125.80 (CH), 126.86 (CH), 127.69 (CH), 127.93 (CH), 131.22 (CH), 132.56 (CH), 135.60 (CH), 135.81 (CH), 137.41 (CH), 139.75 (CH), 140.74 (CH), 154.24 (C ₄=O), 178.86 (C ₂).

35b: ¹H NMR, (CDCl ₃, 300MHz), δ (ppm): 2.93 (4H, s, N-CH ₂-morpholine); 3.50 (4H, s, O-CH ₂-morpholine); 5.69 (1H, s, CH-3); 7.19-7.45 (6H, m, H _Aromatics-diphenylene-oxide and H _Aromatics-7); 7.64 (1H, s, H _Aromatics-8); 7.95 (3H, m, H _Aromatics-diphenylene-oxide); 8.30 (1H, s, H _Aromatics-6).

¹³C-NMR, (CDCl ₃, 300MHz), δ (ppm): 46.87 (CH ₂-N-morpholine); (66.27 CH-O-morpholine); 92.17 (CH-3); 112.10; 121.44; 121.80; 123.43; 123.92; 124.42; 125.82; 126.67; 128.49; 129.11; 133.84; 136.04; 153.32; 154.44; 156.31; 178.93.

Embodiment 3:2-amino-N-[4-(2-morpholine-4-base-4-oxo-4H-pyrido [2,1-a] pyrimidine-9-yl)-dibenzothiophene-1-yl]-ethanamide (36) synthetic

In round-bottomed flask, with 9-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (9) (152.7mg; 0.356mmol) be dissolved among the anhydrous DMA (4.5ml).With triethylamine (109.3 μ L; 0.784mmol) and chloroacetyl chloride (31.03 μ L; 0.392mmol) join in this solution, in room temperature this reactant was stirred 4 hours.The aliquots containig (0.5ml) of reaction product is joined in 9 pipes that contain different amine (3 equivalent) in the greenhouse workstation each.In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC/MS bottle.All LC/MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 4:3-amino-N-[4-(2-morpholine-4-base-4-oxo-4H-pyrido [2,1-a] pyrimidine-9-yl)-dibenzothiophene-1-yl]-propionic acid amide (37) synthetic

In round-bottomed flask, with 9-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-pyrido [2,1-a] pyrimidin-4-ones (23) (127mg; 0.296mmol) be dissolved among the anhydrous DMA (4ml).With triethylamine (90.8 μ L; 0.652mmol) and 3-bromo propionyl chloro (32.8 μ L; 0.326mmol) join in this solution, in room temperature this reactant was stirred 4 hours.The aliquots containig (0.5ml) of reaction product is joined in 8 pipes that contain different amine (3 equivalent) in the greenhouse workstation each.In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC/MS bottle.All LC/MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 5:2-amino-N-[4-(2-morpholine-4-base-4-oxo-1,4-dihydro-quinoline-8-yl)-dibenzothiophene-1-yl]-ethanamide (38) synthetic

In round-bottomed flask, with 8-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-1H-quinoline-4-ketone (25) (153.9mg; 0.36mmol) be dissolved among the anhydrous DMA (4.5ml).With triethylamine (110.3 μ L; 0.696mmol) and chloroacetyl chloride (31.5 μ L; 0.396mmol) join in this solution, in room temperature this reactant was stirred 4 hours.The aliquots containig (0.5ml) of reaction product is joined in 9 pipes that contain different amine (3 equivalent) in the greenhouse workstation each.In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC/MS bottle.All LC/MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 6:3-amino-N-[4-(2-morpholine-4-base-4-oxo-1,4,4a, 8a-tetrahydrochysene-quinoline-8-yl)-dibenzothiophene-1-yl]-propionic acid amide (39) synthetic

In round-bottomed flask, with 8-(1-amino-dibenzothiophene-4-yl)-2-morpholine-4-base-1H-quinoline-4-ketone (25) (79.2mg; 0.185mmol) be dissolved among the anhydrous DMA (3ml).With triethylamine (56.8 μ L); 0.407mmol) and 3-bromo propionyl chloro (20.5 μ L; 0.204mmol) join in this solution, in room temperature this reactant was stirred 4 hours.The aliquots containig (0.5ml) of reaction product is joined in 6 pipes that contain different amine (3 equivalent) in the greenhouse workstation each.In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC/MS bottle.All LC/MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 7:2-amino-N-[4-(2-morpholine-4-base-4-oxo-4H-pyrido [2,1-a] pyrimidine-9-yl)-diphenylene-oxide-1-yl]-ethanamide (40) synthetic

(191mg 0.46mmol) is dissolved among the anhydrous DMA (3.5ml) with 9-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-pyrido [1,2-a] pyrimidin-4-one (27).With triethylamine (129 μ L, 0.92mmol) and chloroacetyl chloride (40 μ L 0.51mmol) join in this solution, in room temperature this reaction mixture are stirred 4 hours.The aliquots containig (0.5ml) of gained solution is joined in 7 different pipes in the greenhouse workstation.Each pipe contains different amine (3 equivalent).In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC-MS bottle.

All LC-MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 8:3-amino-N-[4-(2-morpholine-4-base-4-oxo-4H-pyrido [2,1-a] pyrimidine-9-yl)-diphenylene-oxide-1-yl]-propionic acid amide (41) synthetic

(147mg 0.36mmol) is dissolved among the anhydrous DMA (3ml) with 9-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-pyrido [1,2-a] pyrimidin-4-one (27).(99 μ L, 0.71mmol) (37 μ L 0.39mmol) join in this solution, in room temperature this reaction mixture are stirred 4 hours with the 3-chlorpromazine chloride with triethylamine.The aliquots containig (0.5ml) of gained solution is joined in 6 different pipes in the greenhouse workstation.Each pipe contains different amine (3 equivalent).In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC-MS bottle.All LC-MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 9:2-amino-N-[4-(2-morpholine-4-base-4-oxo-1,4-dihydro-quinoline-8-yl)-diphenylene-oxide-1-yl]-ethanamide (42) synthetic

(126mg 0.31mmol) is dissolved among the anhydrous DMA (3.5ml) with 8-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-1H-quinoline-4-ketone (29).With triethylamine (85 μ L, 0.61mmol) and chloroacetyl chloride (27 μ L 0.34mmol) join in this solution, in room temperature this reaction mixture are stirred 4 hours.The aliquots containig (0.5ml) of gained solution is joined in 7 different pipes in the greenhouse workstation.Each pipe contains different amine (3 equivalent).In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC-MS bottle.All LC-MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 10:3-amino-N-[4-(2-morpholine-4-base-4-oxo-1,4,4a, 8a-tetrahydrochysene-quinoline-8-yl)-diphenylene-oxide-1-yl]-propionic acid amide (43) synthetic

(115mg 0.28mmol) is dissolved among the anhydrous DMA (3ml) with 8-(1-amino-diphenylene-oxide-4-yl)-2-morpholine-4-base-1H-quinoline-4-ketone (29).(78 μ L, 0.56mmol) (29 μ L 0.31mmol) join in this solution, in room temperature this reaction mixture are stirred 4 hours with the 3-chlorpromazine chloride with triethylamine.The aliquots containig (0.5ml) of gained solution is joined in 6 different pipes in the greenhouse workstation.Each pipe contains different amine (3 equivalent).In room temperature with the parallel stirring of reaction mixture 18 hours.With each effective minimum methyl alcohol (1.5ml at most) dilution, transfer in the LC-MS bottle.All LC-MS bottles are carried out QC and half preparation HPLC to carry out purifying.

Embodiment 11:2-amino-N-[4-(2-morpholine-4-base-1-chromene-4-ketone)-diphenylene-oxide-1-yl]-ethanamide (44) synthetic

In 8-(1-amino-diphenylene-oxide-4-the yl)-2-morpholine-4-base-solution (0.02M) of 1-chromene-4-ketone (1 equivalent) in chloroform, add yellow soda ash (2 equivalent), add chloroacetyl chloride (1.1 equivalent) then.In room temperature this mixture was stirred 4 hours, add suitable amine (1.2 equivalent) then.This reactant is heated to 60 ℃ reaches 24 hours, vacuum concentration by the preparation HPLC purifying, obtains required product then then.

Embodiment 12:3-amino-N-[4-(2-morpholine-4-base-1-chromene-4-ketone)-diphenylene-oxide-1-yl]-propionic acid amide (45) synthetic

In 8-(1-amino-diphenylene-oxide-4-the yl)-2-morpholine-4-base-solution (0.02M) of 1-chromene-4-ketone (1 equivalent) in chloroform, add yellow soda ash (2 equivalent), add bromo propionyl chloro (1.1 equivalent) then.In room temperature this mixture was stirred 4 hours, add suitable amine (1.2 equivalent) then.This reactant is heated to 60 ℃ reaches 24 hours, vacuum concentration by the preparation HPLC purifying, obtains required product then then.

Biology embodiment

DNA-PK suppresses

For the vitro inhibition effect of assessing compound, utilize following test determination IC to DNA-PK ₅₀Value.

From HeLa nucleus extract, separate mammiferous DNA-PK (500ng/ml) (Gell according to chromatography, employing Q-agarose, S-agarose with heparin-agarose, D. with Jackson S.P., NucleicAcids Res.27:3494-3502 (1999)).In polypropylene 96 orifice plates, containing 25mM Hepes, pH7.4,12.5mM MgCl ₂, 50mM KCl, 1mM DTT, 10% glycerine, 0.1%NP-40 and 1mg substrate GST-p53N66 (66 amino-acid residues of N-terminal of the people's wild type p53 that merges with glutathione S-transferase) buffer reagent in, with the final volume of 40 μ l in 30 ℃ of activity of measuring DNA-PK (250ng).For test mixture, add different concns inhibitor (final concentration be 1%DMSO in).After hatching 10 minutes, add ATP, make that final concentration is 50 μ M, also add 30 aggressiveness (30mer) double-stranded DNA oligonucleotide (final concentration is 0.5ng/ml) with initiation reaction.After the jolting 1 hour, 150 μ l phosphate buffered saline (PBS)s (PBS) are joined in this reactant, then 5 μ l are transferred to every hole and contain in the 96 hole opaque white color plates of 45 μ l PBS, in every hole, make the GSTp53N66 substrate combine 1 hour with the hole.In order to detect the phosphorylation on Serine 15 residues of p53 that is caused by DNA-PK, (Cell Signaling Technology) is used for basic ELISA method with p53 phosphoserine-15 antibody.The second antibody (Pierce) that will be conjugated with anti-rabbit HRP then is used for this ELISA, adds chemical illuminating reagent (NEN Renaissance) then and comes detection signal, measures this signal by carry out the chemoluminescence counting through TopCountNXT (Packard).

Use following formula to calculate the enzymic activity of each compound then:

The result is hereinafter with IC ₅₀Value (the repressed concentration of 50% enzymic activity) is discussed.These results in the different concns scope, be generally 10 μ M and measure until 0.001 μ M.These IC ₅₀Value value as a comparison is used to differentiate the increase of compound usefulness.

Survival strengthens ratio

It is the ratio that the cell that caused by the DNA-PK inhibitor after 2 gray(Gy)s (Gray) radiation is killed enhancing and unirradiated nonirradiated control cells that survival strengthens than (SER).The DNA-PK inhibitor uses with the fixed concentration of 500nM.By Faxitron 43855D machine, with 1Gy/ minute dose rate generation radiation.Calculate SER under 2 gray(Gy) radiation by following formula:

The degree that survival test (clonogenic survival assay) monitoring cell is killed takes place by the standard clone.In brief, 6 orifice plates of tissue culture treated are inoculated with suitable concentration with the HeLa cell, produced 100-200 colony/hole, it is put back in the incubator, so that cell attachment.After 4 hours, compound or vehicle Control are added to cell.In the presence of inhibitor, cell was hatched 1 hour then, use the box X line of Faxitron 43855D machine to carry out radiation then with 2 gray(Gy)s.Then cell was hatched 16 hours in addition, replace substratum with the fresh culture that does not have the DNA-PK inhibitor subsequently.After 8 days, the colony that forms is fixed, (Sigma, Poole UK), use automatic colony count device (Oxford Optronics Ltd, Oxford, UK) counting to carry out Giemsa staining.Data calculated as mentioned above.

The result

All compound performance DNA-PK suppress active, the IC of its demonstration ₅₀Less than about 500nM.

Demonstrating special usefulness, IC aspect the DNA-PK inhibition ₅₀Less than the compound of about 100nM comprise 23,25,31,34b, 35a-b, 36a-d, 36f-k, 37b-e, 38b, 38d-h, 39d-f, 40a-f, 41a-d, 42b, 42d, 42f, 44a-d, 45a, 45c.

The SER of all compound performances is more than 1 or 1.22,23,24,25,31,36a-k, 37a-c, 37e, 38a-h, 39a-f SER is that the compound more than 2 or 2 comprises following compound:.

Claims (15)

1. formula I compound and isomer thereof, salt, solvate, chemical protected form and prodrug:

Wherein:

A, B and D are selected from respectively:

(i)CH、NH、C；

(ii) CH, N, N; With

(iii)CH、O、C；

Dotted line is illustrated in two two keys on the suitable location;

R ^N1And R ^N2Be independently selected from hydrogen, optional substituted C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl perhaps can form the optional substituted heterocycle with 4 to 8 annular atomses with the nitrogen-atoms that they connect;

If A, B, D are selected from above group (i), (ii), then Z be selected from S, O, C (=O), CH ₂And NH; If A, B, D represent group (iii), then Z be selected from O, C (=O), CH ₂And NH;

R ⁴Be selected from H, OH, NO ₂, NH ₂And Q-Y-X, wherein

Q is-NH-C (=O)-or-O-;

Y is optional substituted C _1-5Alkylene;

X is selected from SR ^S1Or NR ^N3R ^N4, wherein

R ^S1Perhaps R ^N3And R ^N4Be independently selected from hydrogen, optional substituted C _1-7Alkyl, C _5-20Aryl or C _3-20Heterocyclic radical, perhaps R ^N3And R ^N4Can form optional substituted heterocycle with the nitrogen-atoms that they connect with 4 to 8 annular atomses;

If Q is-O-, then X can also be selected from-C (=O)-NR ^N5R ^N6, R wherein ^N5And R ^N6Be independently selected from hydrogen, optional substituted C _1-7Alkyl, C _5-20Aryl or C _3-20Heterocyclic radical, perhaps R ^N5And R ^N6Can form optional substituted heterocycle with the nitrogen-atoms that they connect with 4 to 8 annular atomses, and

If Q is-NH-C (=O)-, then-Y-X can also be selected from C _1-7Alkyl;

Condition is: if A, B, D represent that group (iii) and R ^N1And R ^N2Form morpholino group, then R with their bonded carbon atoms ⁴Cannot be H.

2. according to the compound of claim 1, R wherein ⁴Be Q-Y-X.

3. according to the compound of claim 1 or claim 2, wherein Q be-NH-C (=O)-and X be NR ^N3R ^N4

4. according to the compound of claim 1 or claim 2, wherein Q is-O-, and X is NR ^N3R ^N4, and Y is optional substituted C _1-3Alkylene.

5. according to each compound in the claim 1 to 4, wherein Z takes the circumstances into consideration to be selected from S and O.

6. according to each compound in the claim 1 to 5, wherein R ^N1And R ^N2The nitrogen-atoms that connects with them forms the heterocycle with 4 to 8 atoms.

7. according to each compound in the claim 1 to 5, wherein R ^N1And R ^N2The nitrogen-atoms that connects with their forms and is selected from morpholino base and the parathiazan group for base.

8. composition comprises according to each compound and pharmaceutically useful carrier or thinner in the claim 1 to 7.

9. according to each compound in the claim 1 to 7, be used for the treatment of method.

10. each compound is used for the treatment of by the purposes in the medicine that suppresses the disease that DNA-PK improves in preparation in the claim 1 to 7.

11. each compound is used for the purposes of the medicine of following purpose in the claim 1 to 7 in preparation:

(a) being used as adjuvant or be used in cancer therapy strengthens with the treatment of ionization radiation or the chemotherapeutic effectiveness to tumour cell; Perhaps

(b) disease of treatment retrovirus-mediated method.

12., be used for the treatment of by suppressing the disease that DNA-PK improves according to each compound in the claim 1 to 7.

13., be used for according to each compound in the claim 1 to 7:

(a) being used as adjuvant or be used in cancer therapy strengthens with the treatment of ionization radiation or the chemotherapeutic effectiveness to tumour cell; Perhaps

(b) be used for the treatment of the disease of retrovirus-mediated method.

14. treatment suffers from the method by the curee who suppresses the disease that DNA-PK improves, and comprises to described curee using according to each compound in the claim 1 to 7.

16. in external or body, suppress the method for DNA-PK, comprise making contacting of cell and significant quantity according to each compound in the claim 1 to 7.