HK1181771B - Pharmaceutically active disubstituted triazine derivatives - Google Patents
Pharmaceutically active disubstituted triazine derivatives Download PDFInfo
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- HK1181771B HK1181771B HK13109205.7A HK13109205A HK1181771B HK 1181771 B HK1181771 B HK 1181771B HK 13109205 A HK13109205 A HK 13109205A HK 1181771 B HK1181771 B HK 1181771B
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Description
The present invention relates to disubstituted triazine derivatives and/or pharmaceutically acceptable salts thereof, the use of these derivatives as pharmaceutically active substances, in particular for the prevention and/or treatment of cell proliferative diseases, inflammatory and immunological diseases, cardiovascular diseases and infectious diseases. In addition, the present invention relates to a pharmaceutical composition comprising at least one of said disubstituted triazine derivatives and/or pharmaceutically acceptable salts thereof.
Members of the cyclin-dependent kinase (CDK) family that initiate passage through the cell cycle are considered attractive therapeutic targets, especially for cancer. CDK family members that control other processes such as transcription and RNA processing have received less attention to date, but there is increasing experimental evidence that they are involved in different pathological processes. As a universal transcriptional regulator, CDK9 is a therapeutic target for the treatment of diseases such as inflammation, viral replication (e.g., HIV, EBV, and HCV), cancer, and cardiac hypertrophy.
CDK9 regulates transcription by phosphorylating RNA polymerase II and additional regulatory factors, thereby enabling efficient extension of transcription. Certain gene subgroups are, in particular, genes encoding rapidly renewing RNA or proteins (e.g., immediate early genes of inflammatory response, NF-. kappa.B activating genes (Brasier2008, CellCycle7:17,2661-2666, Hargreaves et al (2009) Cell138, 129-145)); and anti-apoptotic genes (e.g., MCL-1 and Bcl-2 family members) appear to be particularly sensitive to CDK9 inhibition.
In addition, hypertrophic growth of cardiomyocytes has been reported to be associated with CDK9 activation. In addition, viruses such as human immunodeficiency virus automatically recruit CDK9 to nascent RNA transcripts to aid their replication. The dependence of CDK9 activity on expression of anti-apoptotic genes makes it an attractive therapeutic target for various forms of leukemia, such as Chronic Lymphocytic Leukemia (CLL), Acute Myelogenous Leukemia (AML) and acute lymphoblastic leukemia, as well as solid tumors such as prostate, lung, colon, breast and pancreatic cancers. In addition, CDK9 inhibitors were active in a stroke model (Osuga2000, PNAS97(18): 10254-10259).
For reviews see Wang,2009(trends pharmaceutical sciences29:6,302-313), and Kohoutek,2009 (CellVision 2009,4: 19).
It is an object of the present invention to provide compounds and/or pharmaceutically acceptable salts thereof useful as pharmaceutically active substances, particularly for the prevention and/or treatment of cell proliferative diseases, inflammatory diseases, immunological diseases, cardiovascular diseases and infectious diseases, and compositions comprising at least one of those compounds and/or pharmaceutically acceptable salts thereof as pharmaceutically active ingredients.
This object is achieved by: compounds according to independent claim 1 and/or their pharmaceutically acceptable salts, the compounds according to the invention for use as pharmaceutically active substances, the use of the compounds according to the invention according to independent claim 7 for the preparation of pharmaceutical compositions for the prophylaxis and/or treatment of infectious diseases including opportunistic diseases, immunological diseases, autoimmune diseases, cardiovascular diseases, cell proliferative diseases, inflammation, erectile dysfunction and stroke, the use of the compounds according to the invention as inhibitors of the protein kinase CDK 9.
Further advantageous features, aspects and details of the invention are apparent from the dependent claims, the description, the embodiments and the drawings.
The novel disubstituted triazine compounds of the present invention are defined by the general formula (I),
wherein
R1Is that
L is a bond or-CR5R6-、-CR5R6-CR7R8-、-CR5R6-CR7R8-CR9R10-、-CR5R6-CR7R8-CR9R10-CR11R12-；
R5-R12Each independently represents-H, -CH3、-C2H5、-C3H7、-F、-Cl、-Br、-I；
R3Is selected from-H, -NO2、-NH2、-CN、-F、-Cl、-Br、-I、-CH3、-C2H5、-Ph、-C3H7、-CH(CH3)2、-C4H9、-CH2-CH(CH3)2、-CH(CH3)-C2H5、-C(CH3)3、-O-CH3、-O-C2H5、-O-C3H7、-O-CH(CH3)2、-O-C4H9、-O-CH2-CH(CH3)2、-O-CH(CH3)-C2H5、-O-C(CH3)3、-CR13R14R21、-CR13R14CR15R16R21、-O-CR13R14R21、-CR13R14-CR15R16-CR17R18R21、-CR13R14-CR15R16-CR17R18-CR19R20R21、-O-CR13R14-CR15R16R21、-O-CR13R14-CR15R16-CR17R18R21、-SO2R22、-CONR23R24、-NR25COR22、-O-CR13R14-CR15R16-CR17R18-CR19R20R21、-NR25SO2NR23R24、-NR25SO2R22、-NR25CONR23R24、-SO2NR23R24、-SO(NR26)R27、-NH-CO-NH-Ph；
R13-R21、R29-R32And R33-R48Each independently represents-H, -F, -Cl, -Br, -I;
R26is-H, -CH3、-C2H5、-C3H7、-CH(CH3)2、-C4H9、-CH2-CH(CH3)2、-CH(CH3)-C2H5、-C(CH3)3、-C5H11、-CH(CH3)-C3H7、-CH2-CH(CH3)-C2H5、-CH(CH3)-CH(CH3)2、-C(CH3)2-C2H5、-CH2-C(CH3)3、-CH(C2H5)2、-C2H4-CH(CH3)2、-C6H13、-C3H6-CH(CH3)2、-C2H4-CH(CH3)-C2H5、-CH(CH3)-C4H9、-CH2-CH(CH3)-C3H7、-CH(CH3)-CH2-CH(CH3)2、-CH(CH3)-CH(CH3)-C2H5、-CH2-CH(CH3)-CH(CH3)2、-CH2-C(CH3)2-C2H5、-C(CH3)2-C3H7、-C(CH3)2-CH(CH3)2、-C2H4-C(CH3)3、-CH(CH3)-C(CH3)3、-CR13R14R21、-COR28、-CR13R14-CR15R16R21、-CR13R14-CR15R16-CR17R18-CR19R20-CR29R30R21、-CR13R14-CR15R16-CR17R18R21、-CR13R14-CR15R16-CR17R18-CR19R20R21、-CR13R14-CR15R16-CR17R18-CR19R20-CR29R30-CR31R32R21、-COOR28、
These C3-C6-cycloalkyl may be further selected from R by one, two, three, four, five or more33-R48Substituted with the substituent(s);
R22、R27and R28Is independently selected from-CR49R50R51、-CR49R50-CR52R53R51、-CR49R50-CR52R53-CR54R55-CR56R57-CR58R59R51、-CR49R50-CR52R53-CR54R55R51、-CR49R50-CR52R53-CR54R55-CR56R57R51、-CR49R50-CR52R53-CR54R55-CR56R57-CR58R59-CR60R61R51、-CH2Ph, wherein the phenyl groups may be further selected from R by one, two, three, four or five5-R12Is substituted by a substituent of2Ph；
To R26C in the list3-C6-cycloalkyl may be further selected from R by one, two, three, four, five or more33-R48Substituted with the substituent(s);
R49-R61each independently represents-H, -CH3、-C2H5、-C3H7、-C4H9、-F、-Cl、-Br、-I、-OH、-NO2、-NH2；
R23And R24Independently selected from-H, -CR49R50R51、-CR49R50-CR52R53R51、-CR49R50-CR52R53-CR54R55-CR56R57-CR58R59R51、-CR49R50-CR52R53-CR54R55R51、-CR49R50-CR52R53-CR54R55-CR56R57R51、-CR49R50-CR52R53-CR54R55-CR56R57-CR58R59-CR60R61R51、CR49R50-CR52R53-O-R51’、-CR49R50-CR52R53-CR54R55-O-R51’、-CR49R50-CR52R53-NR51’R51”、-CR49R50-CR52R53-CR54R55-NR51’R51”、-CR49R50-CR52R53-CR54R55-CR56R57-NR51’R51”、-CR49R50-CR52R53-CR54R55-CR56R57-CR58R59-NR51’R51", phenyl, substituted phenyl, benzyl, substituted benzyl, or
Residue R23And R24Both of which form, together with the nitrogen atom to which they are attached, an azetidine, pyrrolidine, piperidine, piperazine, azetidine cycloheptane (azepane) or morpholine ring;
R51' and R51"independently of one another represents-H or-CH3、-C2H5、-C3H7、-C4H9、-CH2Ph、-COOC(CH3)3、-COOCH3、-COOCH2CH3、-COOCH2CH2CH3、-COOCH(CH3)2、-COOCH2Ph、-COCH3；
And R is25Is selected from-H, -CH3、-C2H5、-C3H7、-CH(CH3)2、-C4H9、-CH2-CH(CH3)2、-CH(CH3)-C2H5or-C (CH)3)3；
R4Is selected from-H, -NO2、-NH2、-CN、-F、-Cl、-Br、-I、-CONH2、-SO2CH3、-SO2C2H5、-SO2C3H7、-NH-SO2-CH3、-NH-SO2-C2H5、-NH-SO2-C3H7、-NHCO-CH3、-NHCO-C2H5、-NHCO-C3H7、-SO2NR23R24、-CH2-SO2NR23R24、-C2H4-SO2NR23R24、-C3H6-SO2NR23R24、-SO2NH2、-CH2-SO2NH2、-C2H4-SO2NH2、-C3H6-SO2NH2、
-CR62R63R64、-CR62R63-CR65R66-CR67R68-CR69R70R64、-O-CR62R63-CR65R66R64、-O-CR62R63-CR65R66-CR67R68R64、-CR62R63-CR65R66-CR67R68R64、-O-CR62R63-CR65R66-CR67R68-CR69R70R64、-CR62R63-CR65R66R64、-O-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72R64、-O-CR62R63R64、-O-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72-CR73R74R64、-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72R64、-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72-CR73R74R64、-OCH2Ph、
These C3-C6Cycloalkoxy and C3-C6-cycloalkyl may be further selected from R by one, two, three, four, five or more33-R48Substituted with the substituent(s);
R62-R74each independently represents-H, -Ring-C3H5-Ring-C4H7-Ring-C5H9、-CR75R76R77、-CR75R76-CR78R79R77、-CR75R76-CR78R79-CR80R81R77、-CR75R76-CR78R79-CR80R79-CR82R81R77、-F、-Cl、-Br、-I、-Ph；
R75-R82Each independently represents-H, -F, -Cl, -Br, -I, -NH2；
If R is4and-L-R3Ortho-position of (A) is connected, then R4Can be reacted with R22、R23、R24Or R25Together form a radical-CH2CH2-or-CH2CH2CH2-；
R2Is that
R83Selected from-H, -OH, -NO2、-CN、-F、-Cl、-Br、-I、-NR23’R24’、-CF3、-CR62R63R64、-CR62R63-NR23’R24’、-CR62R63-CR65R66R64、-CR62R63-CR65R66-NR23’R24’、-CR62R63-CR65R66-CR67R68R64、-CR62R63-CR65R66-CR67R68-NR23’R24’、-O-CR62R63R64、-O-CR62R63-CR65R66R64、-O-CR62R63-CR65R66-CR67R68R64、-CHO、-CH2OH、-CR23’O、-CH2OR23’；
R23' and R24' independently of one another represents-H, -CH3、-C2H5、-C3H7、-CH(CH3)2、-C4H9、-CH2-CH(CH3)2、-CH(CH3)-C2H5、-C(CH3)3(ii) a - (Ring-C)3H5)；
x is a number from 0 to 3;
b is a bond, -CR86R87-、-CR86R87-CR88R89-、-CR86R87-CR88R89-CR90R91-、-CR86R87-CR88R89-CR90R91-CR92R93-、-CR86R87-CR88R89-CR90R91-CR92R93-CR94R95-、-CR86R87-CR88R89-CR90R91-CR92R93-CR94R95-CR96R97-；
R86-R97Each independently represents-H, -CH3、-C2H5、-C3H7、-C4H9、-F、-Cl、-Br、-I；
Y is a bond, -O-, -S-, -SO2-、-SO2NH-、-NHSO2-、-CO-、-COO-、-OOC-、-CONH-、-NHCO-、-NH-、-N(CH3)-、-NH-CO-NH-、-O-CO-NH-、-NH-CO-O-；
R84Selected from the group consisting of a bond, -CR86R87-、-CR86R87-CR88R89-CR90R91-、-CR86R87-CR88R89-CR90R91-CR92R93-、-CR86R87-CR88R89-CR90R91-CR92R93-CR94R95-、-CR86R87-CR88R89-、-CR86R87-CR88R89-CR90R91-CR92R93-CR94R95-CR96R97-；
R85Is selected from
(i)-H、-OH、-OCH3、-OC2H5、-OC3H7-O-Ring-C3H5、-OCH(CH3)2、-OC(CH3)3、-OC4H9、-Ph、-OPh、-OCH2-Ph、-OCPh3、-SH、-SCH3、-SC2H5、-SC3H7-S-Ring-C3H5、-SCH(CH3)2、-SC(CH3)3、-SC4H9、-NO2、-F、-Cl、-Br、-I、-P(O)(OH)2、-P(O)(OCH3)2、-P(O)(OC2H5)2、-P(O)(OCH(CH3)2)2、-Si(CH3)2(C(CH3)3)、-Si(C2H5)3、-Si(CH3)3、-CN、-CHO、-COCH3、-COC2H5、-COC3H7-CO-Ring-C3H5、-COCH(CH3)2、-COC(CH3)3、-COC4H9、-COOH、-COOCH3、-COOC2H5、-COOC3H7、-COOC4H9-COO-ring-C3H5、-COOCH(CH3)2、-COOC(CH3)3、-OOC-CH3、-OOC-C2H5、-OOC-C3H7、-OOC-C4H9-OOC-Ring-C3H5、-OOC-CH(CH3)2、-OOC-C(CH3)3、-CONR23’R24’、-NHCOCH3、-NHCOC2H5、-NHCOC3H7-NHCO-Ring-C3H5、-NHCO-CH(CH3)2、-NHCOC4H9、-NHCO-C(CH3)3、-NHCO-OCH3、-NHCO-OC2H5、-NHCO-OC3H7-NHCO-O-Ring-C3H5、-NHCO-OC4H9、-NHCO-OCH(CH3)2、-NHCO-OC(CH3)3、-NHCO-OCH2Ph、-NR23R24、-CF3、-SOCH3、-SOC2H5、-SOC3H7-SO-Ring-C3H5、-SOCH(CH3)2、-SOC(CH3)3、-SO2CH3、-SO2C2H5、-SO2C3H7、-SO2-Ring-C3H5、-SO2CH(CH3)2、-SO2C4H9、-SO2C(CH3)3、-SO3H、-SO2NR23R24、-OCF3、-OC2F5、-O-COOCH3、-O-COOC2H5、-O-COOC3H7-O-COO-Ring-C3H5、-O-COOC4H9、-O-COOCH(CH3)2、-O-COOCH2Ph、-O-COOC(CH3)3、-NH-CO-NH2、-NH-CO-NHCH3、-NH-CO-NHC2H5、-NH-CO-NHC3H7、-NH-CO-NHC4H9-NH-CO-NH-Ring-C3H5、-NH-CO-NH[CH(CH3)2]、-NH-CO-NH[C(CH3)3]、-NH-CO-N(CH3)2、-NH-CO-N(C2H5)2、-NH-CO-N(C3H7)2、-NH-CO-N(C4H9)2-NH-CO-N (Ring)-C3H5)2、-NH-CO-N[CH(CH3)2]2、-NH-CO-N[C(CH3)3]2、-NH-C(=NH)-NH2、-NH-C(=NH)-NHCH3、-NH-C(=NH)-NHC2H5、-NH-C(=NH)-NHC3H7、-NH-C(=NH)-NHC4H9-NH-C (= NH) -NH-ring-C3H5、-OCH2-Ring-C3H5、-NH-C(=NH)-NH[CH(CH3)2]、-NH-C(=NH)-NH[C(CH3)3]、-NH-C(=NH)-N(CH3)2、-NH-C(=NH)-N(C2H5)2、-NH-C(=NH)-N(C3H7)2-NH-C (= NH) -N (cyclo-C)3H5)2、-NH-C(=NH)-N(C4H9)2、-NH-C(=NH)-N[CH(CH3)2]2、-NH-C(=NH)-N[C(CH3)3]2、-O-CO-NH2、-O-CO-NHCH3、-O-CO-NHC2H5、-O-CO-NHC3H7、-O-CO-NHC4H9-O-CO-NH-Ring-C3H5、-O-CO-NH[CH(CH3)2]、-O-CO-NH[C(CH3)3]、-O-CO-N(CH3)2、-O-CO-N(C2H5)2、-O-CO-N(C3H7)2、-O-CO-N(C4H9)2-O-CO-N (Ring-C)3H5)2、-O-CO-N[CH(CH3)2]2、-O-CO-N[C(CH3)3]2；
(ii) An aromatic or heteroaromatic monocyclic or bicyclic ring selected from: 2-thienyl, 3-thienyl, 2-furyl, 3-furyl, 2-oxazolyl, 3-oxazolyl, 4-oxazolyl, 2-thiazolyl, 3-thiazolyl, 4-thiazolyl, 1-pyrazolyl, 3-pyrazolyl, 4-pyrazolyl, 5-pyrazolyl, 1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, phenyl, 1-naphthyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl, 4-pyrazinyl, 1,3, 5-triazin-2-yl, 2-thiazolyl, 3-thiazolyl, 4-pyrazolyl, 1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 4-pyrazinyl, 1,3, 5-triazin-2-yl, and the like,
Optionally substituted by one or two groups selected from-F, -Cl, -Br, -I, -OCH3、-CH3、-NO2、-CN、-CF3Substituted with the substituent(s);
(iii) a saturated ring selected from: cyclopentyl, azetidin-1-yl,
R99represents-H, -CH3、-CH2Ph、-COOC(CH3)3、-COOCH3、-COOCH2CH3、-COOCH2CH2CH3、-COOCH(CH3)2、-COOCH2Ph、-COCH3；
If R is83With a radical-B-Y-R84-R85Ortho-position of (a) is linked, then-B-Y-R84-R85May be substituted by a substituent R83Together form the group-OCH2O-；
With the proviso that if the group-B-Y-R84-R85Is hydrogen, then R83Is not hydrogen;
R98is selected from-NO2、-CN、-F、-Cl、-Br、-I、-NH2、-OH、-CR62R63-CR65R66-CR67R68-CR69R70R64、-O-CR62R63R64、-O-CR62R63-CR65R66R64、-O-CR62R63-CR65R66-CR67R68R64、-O-CR62R63-CR65R66-CR67R68-CR69R70R64、-O-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72R64、-CR62R63-CR65R66-CR67R68R64、-O-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72-CR73R74R64、-CR62R63-CR65R66R64、-CR62R63-O-CR65R66-CR67R68-CR69R70R64、-CR62R63-O-CR65R66-CR67R68R64、-CR62R63-O-CR65R66-CR67R68-CR69R70-CR71R72R64、-CR62R63-O-CR65R66R64、-CR62R63-O-CR65R66-CR67R68-CR69R70-CR71R72-CR73R74R64、-CR62R63R64、-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72R64、-OCH2Ph、-OCH2-CH2-Ph、-CH2-O-CH2-Ph、-CR62R63-CR65R66-CR67R68-CR69R70-CR71R72-CR73R74R64(ii) a With the proviso that if R98Not amino in para position to the bond between the pyridine and triazine ring, then R98Ortho-to the bond between the pyridine and the triazine ring;
R100is selected from-H, -NO2、-CN、-F、-Cl、-Br、-I、-NH2、-OH、-CF3、-CH3、-CH2CH3、-CH2CH2CH3、-CH(CH3)2、-OCH3、-OCH2CH3、-OCH2CH2CH3、-OCH(CH3)2、-OCF3、-OCH2Ph；
And with the proviso that if R1Is phenyl and R2Is also phenyl, then only R is allowed1On the phenyl radical or R2The presence of a chlorine substituent on the phenyl group rather than both groups;
and with the proviso that the compound 4- [4- (2-benzoylaminophenyl) - [1,3,5] triazin-2-ylamino ] benzamide is excluded;
and enantiomers, stereoisomeric forms, mixtures of enantiomers, diastereomers, mixtures of diastereomers, prodrugs, hydrates, solvates, acid salt forms, tautomers and racemates of the aforementioned compounds and pharmaceutically acceptable salts or solvate salts thereof.
The expression prodrug is defined as a substance that is administered in an inactive form or in a significantly less active form. Upon administration or incorporation, the prodrug is metabolized in vivo to an active compound.
The expression tautomer is defined as an organic compound that can be interconverted by a chemical reaction called tautomerization. Tautomerization may preferably be catalyzed by bases or acids or other suitable compounds.
Preferably having the general formula (I):
wherein
R1To represent
Wherein
L is a bond, -CH2-、-CH2CH2-or-CF2-, particularly preferably-CH2-；
R3is-SO2NH2、-SO2NH(CH3)、-SO2N(CH3)2、-SO2NH(CH2CH2OCH3)、-NHSO2CH3、-NHSO2CH2CH3、-NHSO2CH2CH2CH3、-NHSO2CF3、-SO2CH3、-NHSO2NH2、-SO(NH)CH3Particularly preferred is-SO2NH2；
R4is-H, -CH3-F, -Cl or-CF3Particularly preferably-H;
R2to represent
Wherein the group-B-Y-R84-R85is-OCH3、-OCH2CH3、-OCH2CH2CH3、-OCH2CH2CH2CH3、-OCH(CH3)2、-OPh、-OCH2Ph、-OCH2(4-pyridyl group), particularly preferably-OCH3；
R83is-H, -F or-Cl;
x is 0, 1 or 2;
R98is-OCH3And R is100is-H, with the proviso that R98Ortho to the bond between the pyridine and the triazine ring.
In a more preferred compound of formula (I),
substituent-L-R3is-SO2NH2、-CH2SO2NH2、-CH2CH2SO2NH2、-CF2SO2NH2、-NHSO2NH2、-CH2NHSO2NH2、-SO2CH3、-SO(NH)CH3、-CH2SO(NH)CH3；
And R is4is-H;
R2is 2-methoxyphenyl, 4-fluoro-2-methoxyphenyl or 6-fluoro-2-methoxyphenyl.
Preference is given to compounds of the formula (I) in which R1Is thatAnd wherein L is a bond or is-CH2-or-CH2CH2-; and R is3Have the meaning defined herein, and more preferably, R3represents-SO2R22or-SO2NR23R24Wherein R is22、R23And R24Have the meaning defined herein, and preferably R22、R23And R24 independently of one another represent-H, -CF3、-CH3、-CH2CH3、-CH2CH2CH3、-CH2CH2CH2CH3、-CH(CH3)2、-CH2-NH2、-CH2-CH2-NH2、-CH2-CH2-CH2-NH2、-CH2-CH2-CH2-CH2-NH2、-CH2-NH-CO-O-C(CH3)3、-CH2-CH2-NH-CO-O-C(CH3)3、-CH2-CH2-CH2-NH-CO-O-C(CH3)3、-CH2-CH2-CH2-CH2-NH-CO-O-C(CH3)3。
Also preferred are compounds of formula (I) wherein L is a bond, -CH2-、-CH2CH2-、-CH2CH2CH2-or-CF2-, more preferably-CH2-or-CH2CH2-。
Preference is given to compounds of the formula (I) in which R2Is that
If the residue R is2Is a benzene ring, the substituents B-Y-R which are in the ortho position to the triazine core bond are preferred84-R85Is not hydrogen, and if the substituent is hydrogen, R83Other than hydrogen, and, in addition, at least one substituent R83Ortho to the triazine core bond. Thus, B-Y-R84-R85And R83One substituent in (a) needs to be different from hydrogen so that R is different from hydrogen2Not an unsubstituted benzene ring. Otherwise, if B, Y and R84Is a bond and R83Different from hydrogen, R is preferably85Is not-H. If two substituents are present, it is preferred that the second substituent is meta to the triazine core bondOr para. If a third substituent is present, a substitution pattern of 2,3,5 or 2,3,4 is preferred. Fluorine is a preferred second and/or third substituent and is preferably located meta or para to the triazine core bond. Therefore, the following R is preferred2Residue:
if R is2The residue is a pyridine ring, R is preferred98One substituent of (a) is located ortho to the bond of the triazine core. Preferably the following R2Residue:
preference is also given to compounds of the formula (I) in which R is85Is that
R3Preferably selected from-H, -NO2、-NH2、-CN、-F、-Cl、-Br、-I、-CH3、-C2H5、-Ph、-C3H7、-CH(CH3)2、-C4H9、-CH2-CH(CH3)2、-CH(CH3)-C2H5、-C(CH3)3、-O-CH3、-O-C2H5、-O-C3H7、-O-CH(CH3)2、-O-C4H9、-O-CH2-CH(CH3)2、-O-CH(CH3)-C2H5、-O-C(CH3)3、-SO2R22and-SO2NR23R24。
R26Preferably selected from-H, -CH3、-C2H5、-C3H7、-CH(CH3)2、-C4H9、-CH2-CH(CH3)2、-CH(CH3)-C2H5、-C(CH3)3、-C5H11、-CH(CH3)-C3H7、-CH2-CH(CH3)-C2H5、-CH(CH3)-CH(CH3)2、-C(CH3)2-C2H5、-CH2-C(CH3)3、-CH(C2H5)2、-C2H4-CH(CH3)2、-C6H13-Ring-C3H5-Ring-C4H7and-Ring-C5H9。
In addition, in the compounds of the general formula (I), R22、R23、R24、R27And R28Each independently selected from-H, -CH3、-C2H5、-C3H7、-C4H9or-CH2Ph。
Preferably, R62-R74Each independently represents-H, -Ph, -Ring-C3H5-Ring-C4H7、-CH3、-C2H5、-C3H7、-C4H9-Ring-C5H9-F, -Cl, -Br or-I.
Preference is also given to compounds of the formula (I) in which R is4Is selected from-H, -NO2、-NH2-CN, -F, -Cl, -Br, -I, -cyclo-C3H5-Ring-C4H7-Ring-C5H9、-CH3、-C2H5、-C3H7、-CH(CH3)2、-C4H9、-CONH2、-SO2CH3、-SO2C2H5、-SO2C3H7、-NH-SO2-CH3、-NH-SO2-C2H5、-NH-SO2-C3H7、-NHCO-CH3、-NHCO-C2H5、-NHCO-C3H7、-SO2NR23R24、-CH2-SO2NR23R24、-C2H4-SO2NR23R24、-C3H6-SO2NR23R24、-SO2NH2、-CH2-SO2NH2、-C2H4-SO2NH2、-C3H6-SO2NH2、
-CH(CH3)-C2H5、-C(CH3)3、-C5H11、-CH2-CH2-CH2-CH2R64、-O-CH2-CH2R64、-CH2R64、-O-CH2-CH2-CH2R64、-CH2-CH2-CH2R64、-O-CH2-CH2-CH2-CH2R64、-CH2-CH2R64、-O-CH2-CH2-CH2-CH2-CH2R64、-CH2-CH2-CH2-CH2-CH2R64、-O-CH2-CH2-CH2-CH2-CH2-CH2R64、-CH2-CH2-CH2-CH2-CH2-CH2R64、-OCH2Ph、-O-CH2R64Wherein R is64represents-Ph, -F, -Cl, -Br or-I. Preference is given to compounds in which R is4Is selected from-NO2、-NH2、-CONH2、-SO2CH3、-SO2C2H5、-SO2C3H7、-NH-SO2-CH3、-NH-SO2-C2H5、-NH-SO2-C3H7、-NHCO-CH3、-NHCO-C2H5、-NHCO-C3H7、-SO2NR23R24、-CH2-SO2NR23R24、-C2H4-SO2NR23R24、-C3H6-SO2NR23R24、-SO2NH2、-CH2-SO2NH2、-C2H4-SO2NH2、-C3H6-SO2NH2、
In addition, two substituents-L-R are particularly preferred3and-R4Not all are hydrogen. Thus, the phenyl substituent R is preferred1And a pyridyl substituent R1Having at least one substituent, and preferably one substituent in the meta position, most preferably the above-mentioned-L-R3and-R4Is in the meta position, and particularly preferably-R mentioned above4Is in the meta position. Therefore, the following R is preferred1The residue, and particularly preferably the following has-L-R3and-R4Substituent R of preferred substituents of1：
Preference is also given to compounds of the formula (I) in which R is83is-H, -OH, -NO2、-CN、-F、-Cl、-Br、-I、-NH2、-NH(CH3)、-N(CH3)2、-NH(C2H5)、-N(C2H5)2、-CF3、-CH3、-C2H5、-C3H7、-CH(CH3)2、-C4H9、-C(CH3)3、-CH2-NH2、-CH2-NH(CH3)、-CH2-N(CH3)2、-CH2-NH(C2H5)、-CH2-N(C2H5)2、-CH2-CH2-NH2、-CH2-CH2-NH(CH3)、-CH2-CH2-N(CH3)2、-CH2-CH2-NH(C2H5)、-CH2-CH2-N(C2H5)2、-CH2-CH2-CH2-NH2、-CH2-CH2-CH2-NH(CH3)、-CH2-CH2-CH2-N(CH3)2、-CH2-CH2-CH2-NH(C2H5)、-CH2-CH2-CH2-N(C2H5)2、-O-CH3、-O-CH2-CH3、-O-CH2-CH2-CH3、-CHO、-CH2OH、-CO-CH3、-CO-CH2-CH3、-CO-CH2-CH2-CH3、-CO-CH2-CH2-CH2-CH3、-CH2O-CH3、-CH2O-CH2-CH3、-CH2O-CH2-CH2-CH3、-CH2F、-CH2Cl、-CH2Br、-CH2-CH2F、-CH2-CH2Cl、-CH2-CH2Br、-CH2-CH2-CH2F、-CH2-CH2-CH2Cl、-CH2-CH2-CH2Br。
Furthermore preferred are compounds of the formula (I) in which B represents a bond, -CH2-、-CH2CH2-、-CH2CH2CH2-、-CH2CH2CH2CH2-、-CH2CH2CH2CH2CH2-and/or wherein Y represents a bond, -O-or-NH-.
Preference is furthermore given to compounds of the formula (I) in which R84Represents a bond, -CH2-、-CH2CH2-、-CH2CH2CH2-、-CH2CH2CH2CH2-、-CH2CH2CH2CH2CH2-。
Preference is also given to compounds of the formula (I) in which R is85is-H, -OH, -OCH3、-OC2H5、-OC3H7-O-Ring-C3H5、-OCH(CH3)2、-OC(CH3)3、-OC4H9、-Ph、-OPh、-OCH2-Ph、-OCPh3、-NO2、-F、-Cl、-Br、-I、-CN、-CHO、-COCH3、-COC2H5、-COC3H7-CO-Ring-C3H5、-COCH(CH3)2、-COC(CH3)3、-COC4H9、-COOH、-COOCH3、-COOC2H5、-COOC3H7、-COOC4H9-COO-ring-C3H5、-COOCH(CH3)2、-COOC(CH3)3、-OOC-CH3、-OOC-C2H5、-OOC-C3H7、-OOC-C4H9-OOC-Ring-C3H5、-OOC-CH(CH3)2、-OOC-C(CH3)3、-CONR23'R24'、-NHCOCH3、-NHCOC2H5、-NHCOC3H7-NHCO-Ring-C3H5、-NHCO-CH(CH3)2、-NHCOC4H9、-NHCO-C(CH3)3、-NHCO-OCH3、-NHCO-OC2H5、-NHCO-OC3H7-NHCO-O-Ring-C3H5、-NHCO-OC4H9、-NHCO-OCH(CH3)2、-NHCO-OC(CH3)3、-NHCO-OCH2Ph、-NR23R24、-CF3、-SOCH3、-SOC2H5、-SOC3H7-SO-Ring-C3H5、-SOCH(CH3)2、-SOC(CH3)3、-SO2CH3、-SO2C2H5、-SO2C3H7、-SO2-Ring-C3H5、-SO2CH(CH3)2、-SO2C4H9、-SO2C(CH3)3、-SO3H、-SO2NR23’R24’、-OCF3、-OC2F5、-NH-CO-NH2、-NH-CO-NHCH3、-NH-CO-NHC2H5、-NH-CO-NHC3H7、-NH-CO-NHC4H9-NH-CO-NH-Ring-C3H5、-NH-CO-NH[CH(CH3)2]、-NH-CO-NH[C(CH3)3]、-NH-CO-N(CH3)2、-NH-CO-N(C2H5)2、-NH-CO-N(C3H7)2、-O-CO-NH2、-O-CO-NHCH3、-O-CO-NHC2H5、-O-CO-NHC3H7、-O-CO-NHC4H9-O-CO-NH-Ring-C3H5、-O-CO-NH[CH(CH3)2]、-O-CO-NH[C(CH3)3]、-O-CO-N(CH3)2、-O-CO-N(C2H5)2、-O-CO-N(C3H7)2、-O-CO-N(C4H9)2-O-CO-N (Ring-C)3H5)2、-O-CO-N[CH(CH3)2]2、-O-CO-N[C(CH3)3]22-thienyl, 3-thienyl, 2-furyl, 3-furyl, 2-oxazolyl, 3-oxazolyl, 4-oxazolyl, 2-thiazolyl, 3-thiazolyl, 4-thiazolyl, 1-pyrazolyl, 3-pyrazolyl, 4-pyrazolyl, 5-pyrazolyl, 1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, phenyl, 1-naphthyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl, 4-pyrazinyl, 1,3, 5-triazin-2-yl, 2-thiazolyl, 3-pyrazolyl, 1-imidazolyl, 2-imidazolyl, 5-imidazolyl, phenyl, 1-naphthyl, 2-pyridyl, 3-pyridyl, 4-pyrimidinyl, 5,
With the proviso that if the group-B-Y-R84-R85Is hydrogen, then R83Is not-H.
Preference is also given to compounds of the formula (I) in which R is98is-NO2、-CN、-F、-Cl、-Br、-I、-NH2、-OH、-CF3、-CH3、-C2H5、-C3H7、-CH(CH3)2、-C4H9、-C(CH3)3、-CH2-NH2、-CH2-NH(CH3)、-CH2-N(CH3)2、-CH2-NH(C2H5)、-CH2-N(C2H5)2、-CH2-CH2-NH2、-CH2-CH2-NH(CH3)、-CH2-CH2-N(CH3)2、-CH2-CH2-NH(C2H5)、-CH2-CH2-N(C2H5)2、-CH2-CH2-CH2-NH2、-CH2-CH2-CH2-NH(CH3)、-CH2-CH2-CH2-N(CH3)2、-CH2-CH2-CH2-NH(C2H5)、-CH2-CH2-CH2-N(C2H5)2、-O-CH3、-O-CH2-CH3、-O-CH2-CH2-CH3、-CH2O-CH3、-CH2O-CH2-CH3、-CH2O-CH2-CH2-CH3、-CH2F、-CH2Cl、-CH2Br、-CH2-CH2F、-CH2-CH2Cl、-CH2-CH2Br、-CH2-CH2-CH2F、-CH2-CH2-CH2Cl、-CH2-CH2-CH2Br、-OCH2Ph、-OCH2-CH2-Ph、-CH2-O-CH2-Ph。
Particular preference is given to compounds of the formula (I) in which
R1Is that
L is a bond, -CH2-or-CH2CH2-；
R3is-H, -SO2NR23R24、-CONR23R24、-NO2、-NH2、-NHSO2R22、-NHCOR22、-SO2R22-NH-CO-NH-Ph or-Ph;
R4is-H, -CH2-SO2NR23R24、-SO2NR23R24、-CONH2、-C2H4-SO2NR23R24、-NH-SO2-CH3、-NH-SO2-C2H5、-NH-SO2-C3H7、-NHCO-CH3、-NHCO-C2H5、-NO2、-NH2、-SO2CH3Or
R23And R24Independently selected from-H, -CH3、-C2H5、-C3H7- (Ring-C)3H5)、-CH2-CH2-CH2-CH2-NH2or-CH2-CH2-CH2-CH2-NH-COOC(CH3)3，
R2To represent
B is a bond or-CH2-；
Y is a bond, -O-, or-NH-;
R83selected from the group consisting of-H, -CN, -F, -Cl, -O-CR62R63R64、-CF3、-CH2OR23’、-CR23’O、-CR62R63-NR23’R24’、-CR62R63R64；
R23' and R24' independently of one another represents-H, -CH3- (Ring-C)3H5)；
R62-R64Each independently represents-H, -CH3-Ph, -F, - (Ring-C)3H5)；
R84Is selected from the group consisting of a bond, -CH2-or-CH2-CH2-CH2-CH2-；
R85Selected from-H, -CF3、-OCH3、-OCH(CH3)2、-CN、-NHCOCH3、-OCH2- (Ring-C)3H5)、-NR23R24、-Ph、-OPh、-NHCO-OC(CH3)3、
R98represents-OCH3；
And salts, solvates or solvates of the aforementioned compounds, in particular the hydrochloride or trifluoroacetate salts of these compounds.
In addition, particular preference is given to compounds of the formula (I) in which
R1Is that
L is a bond, -CH2-or-CH2CH2-；
R3is-H, -SO2NH2、-CONH2、-NO2、-NH2、-NH-SO2-CH3、-NH-SO2-C3H7、-NHCO-CH3、-SO2CH3、-Ph、-SO2-NH-CH2-CH2-CH2-CH2-NH-COOC(CH3)3-NH-CO-NH-Ph or-SO2-NH-CH2-CH2-CH2-CH2-NH2；
R4is-H, -CH2-SO2NH2、-SO2NH2、-C2H4-SO2NH2、-CONH2、-NH-SO2-CH3、-NH-SO2-C3H7、-NHCO-CH3、-NO2、-NH2、-SO2CH3Or is
R2To represent
B is a bond or-CH2-；
Y is a bond, -O-, or-NH-;
R83selected from-H, -F, -Cl, -O-CH3、-O-C2H5、-OCH2- (Ring-C)3H5)、-CN、-CF3、-CH2OH、-CHO、-CH2-NH (Ring-C)3H5)、-CH2-NH(CH3)、-CF3；
R84Is selected from the group consisting of a bond, -CH2-or-CH2-CH2-CH2-CH2-；
R85Selected from-H, -CF3、-OCH3、-OCH(CH3)2、-CN、-NHCOCH3、-OCH2- (Ring-C)3H5)、-NH2-NH- (Ring-C)3H5)、-Ph、-OPh、-NHCO-OC(CH3)3、
R98represents-OCH3；
And salts, solvates or solvates of the aforementioned compounds, in particular the hydrochloride or trifluoroacetate salts of these compounds.
In a particularly preferred embodiment, the present invention relates to compounds of the formula (I) or their salts, solvates or salts of solvates, in particular the hydrochloride or trifluoroacetate salt, where
R1To represent
Wherein the substituent-L-R3is-SO2NH2or-CH2SO2NH2；
R4is-H;
R2represents a 2-methoxyphenyl group, a 4-fluoro-2-methoxyphenyl group or a 2-benzyloxyphenyl group.
In another particularly preferred embodiment, the present invention relates to a compound of formula (I) or a salt, solvate or salt of a solvate thereof, in particular a hydrochloride or trifluoroacetate salt, selected from: 3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide (B1), 3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide (C1), 3- [ (4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide (B2), 3- [ (4- (2-benzyloxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide (B13).
In another particularly preferred embodiment, the present invention relates to 3- [ (4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide, or a salt, solvate or salt of a solvate thereof, particularly the hydrochloride or trifluoroacetate salt.
In another particularly preferred embodiment, the present invention relates to 1- (3- { [4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl ] amino } phenyl) methanesulfonamide hydrochloride.
In which-R is excluded from the invention4and-L-R3These compounds are methoxy or ethoxy.
Compounds in which R is1Is thatAnd wherein R2Is thatAnd wherein-R4and-L-R3One of which is a chlorine substituent, and wherein B-Y-R84-R85and-R83One of which is also a chlorine substituent. More generally, compounds of formula (I) having two or more chlorine substituents are not preferred and may be excluded.
If the group B-Y-R84-R85Represents a substituent-NH-CO-Ph, then phenyl R1Having at least one group not located in the phenyl radical R1And a triazine ring or substituent-L-R3A substituent para to the bond between, wherein L is a bond and is substituted with a substituent-CO-NH2Different. In addition, the following compounds are excluded from the scope of the present invention by disclaiming the claims: 4- [4- (2-benzoylaminophenyl) - [1,3,5]]Triazin-2-ylamino]Benzamide derivatives
In another aspect of the invention, the novel compounds of formula (I) represent chiral compounds. The novel compounds of the general formula (I) represent racemates or S or R enantiomers or mixtures of isomers.
In another preferred embodiment of the present invention, the compounds of general formula (I) are selected from the compounds listed in table 1 below.
TABLE 1
The compounds of the present invention may form salts with organic or inorganic acids or bases. Examples of suitable acids for forming such acid addition salts are hydrochloric, hydrobromic, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, p-aminosalicylic, malic, fumaric, succinic, ascorbic, maleic, sulfonic, phosphonic, perchloric, nitric, formic, propionic, gluconic, lactic, tartaric, hydroxymaleic, pyruvic, phenylacetic, benzoic, p-aminobenzoic, p-hydroxybenzoic, methanesulfonic, ethanesulfonic, nitrous, isethionic, ethylenedisulfonic, p-toluenesulfonic, naphthalenesulfonic, sulfanilic, camphorsulfonic, cerotic (chinoacid), mandelic, o-methylmandelic, hydrobenzenesulfonic (hydrozenesulfonic), picric, adipic, d-o-tolyltartaric, tartronic, (o, m, p) -methyltoluic, naphthylaminesulfonic, salicylic, pyruvic, succinic, fumaric, maleic, fumaric, maleic, fumaric, maleic, fumaric, Trifluoroacetic acid and other inorganic acids or carboxylic acids well known to those skilled in the art. Salts are prepared in a conventional manner by contacting the free base form with a sufficient amount of the desired acid. Preference is given to the methanesulfonate, hydrochloride and trifluoroacetate, particular preference to the trifluoroacetate and hydrochloride.
In the case where the compounds of the invention carry an acidic group, salts may also be formed with inorganic or organic bases. Examples of suitable inorganic or organic bases are, for example, NaOH, KOH, NH4OH, tetraalkylammonium hydroxide, lysine or arginine, and the like. The salts may be prepared in a conventional manner using methods well known in the art, for example by treating a solution of the compound of formula (I) with a solution of an acid selected from the group described above. Synthesis of Compounds
The synthesis of the inventive disubstituted triazines of the present invention is preferably carried out according to the general synthetic sequence shown in schemes 1-3.
Route 1
In a first step, 2, 4-dichloro-1, 3, 5-triazine is reacted with aniline R1NH2Reacting to obtain the 2-arylamino-4-chloro-1, 3, 5-triazine. The reaction is carried out with one equivalent of aniline in an inert solvent (e.g. DMF, THF, DME, dioxane) or an alcohol (e.g. isopropanol) or a mixture of such solvents. The reaction is preferably carried out at a temperature below room temperature so that the reaction mixture remains homogeneous. Preferred conditions use an additional base such as triethylamine or N, N-diisopropylethylamine.
In a second step, the intermediate 2-arylamino-4-chloro-1, 3, 5-triazine is reacted with a boronic acid derivative R2-B(OR)2Reacting to obtain the compound of formula (I). The boronic acid derivative may be boronic acid (R = -H) or an ester of a boronic acid such as isopropyl borate (R = -CH (CH)3)2) Preference is given to esters derived from pinacol (where the boronic acid intermediate forms 2-aryl-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan (R-R = -C (CH))3)2-C(CH3)2-)). Preferably, the two R's each independently represent hydrogen or an alkyl chain having 1 to 10 carbon atoms or a cycloalkyl chain having 3 to 12 carbon atoms, or the two residues R together represent a residue derived from pinacol. The coupling reaction is catalyzed by a Pd catalyst, e.g., a Pd (0) catalyst such as tetrakis (triphenylphosphine) palladium (0) [ Pd (PPh)3)4]Tris (dibenzylideneacetone) dipalladium (0) [ Pd ]2(dba)3]Or using a Pd (II) catalyst such as dichlorobis (triphenylphosphine) palladium (II) [ Pd (PPh)3)2Cl2]Palladium (II) acetate and triphenylphosphine, or more preferably [1, 1' -bis (diphenylphosphino) ferrocene]Palladium dichloride. Preferably in a mixture of a solvent such as dioxane, DMF, DME, THF or isopropanol with water and in a solvent such as sodium bicarbonate or K3PO4The reaction is carried out in the presence of a base in aqueous solution.
Route 2
The synthesis of 1,3, 5-triazines of formula (I) starting from 2, 4-dichloro-1, 3, 5-triazine can be carried out in the reverse reaction sequence compared to scheme 1, so that in the first step the triazine is reacted with a boronic acid derivative, and then in the second step the intermediate triazine is reacted with aniline. The preferred conditions for the coupling reaction of the first step are in the presence of sodium or potassium carbonate as a base in the presence of palladium (II) dichlorobis (triphenylphosphonium) containing [ Pd (PPh) as a catalyst3)2Cl2]The reaction mass was heated in toluene.
Route 3
Chem can pass j.60(1995) 8428-8430 by the methods described in formula (I). The primary amide R2-CONH2With acetals (Yb)The dialkyl acetal of N, N-dimethylformamide, preferably its dimethyl acetal or diethyl acetal, especially with dimethyl acetal (R = -CH)3) Are heated together). Without isolation of the intermediate N-acylformamidine and subsequent reaction with guanidine R1-NH-C(NH)NH2Heated together to convert them into the 1,3, 5-triazines of formula (I). Preferably, the reaction is carried out by heating the reaction mass in dioxane in the presence of a base such as potassium tert-butoxide.
Standard reactions known to those skilled in the art can be used, by reacting with the aromatic ring R1And/or R2The attached substituents are converted to other substituents to prepare several compounds of formula (I). For example, the nitro group may be reduced to an amino group which may be converted to a sulfonamide by reaction with a sulfonyl chloride, to a carboxamide by reaction with phosgene or another activated derivative of a carboxylic acid, to a urea by reaction with an isocyanate. The carbamate substituent can be cleaved to the amino group, particularly by reaction with an acid such as trifluoroacetic acid or hydrochloric acid to cleave the tert-butyl carbamate. Formyl groups can be converted to aminomethyl groups by reaction with primary amines under reductive amination conditions.
Application method
In another aspect of the invention, the novel compounds of the general formula (I) are used as pharmaceutically active substances.
Further aspects of the present invention relate to the use of the compounds of general formula (I) for the preparation of pharmaceutical compositions for the prevention and/or treatment of infectious diseases including opportunistic diseases, immune diseases, autoimmune diseases, cardiovascular diseases, cell proliferative diseases, inflammation, erectile dysfunction and stroke.
The compounds of the invention are useful for inhibiting the activity or expression of CDK 9. Accordingly, it is expected that compounds of formula (I) may be useful as valuable therapeutic agents and, accordingly, in another embodiment, the present invention provides a method of treating a condition associated with or mediated by CDK9 activity in a patient in need of such treatment which comprises administering to said patient an effective amount of a compound of formula (I) as defined above. In certain embodiments, the disorder associated with CDK9 activity is a cell proliferative disorder, particularly cancer.
The term "treating" as used throughout this document is used in a conventional sense, e.g., to monitor or care for an individual to combat, alleviate, reduce, alleviate, ameliorate the condition of a disease or disorder, e.g., cancer.
The term "subject" or "patient" includes an organism, such as a human or non-human animal, capable of suffering from a cell proliferative disorder or a disorder associated with reduced or insufficient programmed cell death (apoptosis) or capable of benefiting from administration of a compound of the invention. Preferred humans include human patients suffering from or susceptible to a cell proliferative disorder or related disorder as described herein. The term "non-human patient" includes vertebrates, such as mammals, e.g., non-human primates, sheep, cows, dogs, cats, and rodents, e.g., mice, as well as non-mammals, e.g., chickens, amphibians, reptiles, and the like.
The term "a disorder associated with or mediated by CDK 9" shall include diseases associated with CDK9 activity or involving CDK9 activity (e.g., CDK9 is overactive) and disorders associated with such diseases. Examples of "disorders associated with or mediated by CDK 9" include disorders arising from increased CDK9 activity due to mutation of genes modulating CDK9 activity, such as LARP7, HEXIM1/2 or 7sksnRNA, or increased CDK9 activity due to activation of the CDK 9/cyclin T/RNA polymerase II complex by viral proteins such as HIV-TAT or HTLV-TAX, or increased CDK9 activity due to activation of the mitogenic signal transduction pathway.
The term "overactive with CDK 9" refers to an increase in the enzymatic activity of CDK9 as compared to normal, non-diseased cells, or which directs an increase in CDK9 activity resulting in reduced or insufficient undesired cell proliferation or programmed cell death (apoptosis), or a mutation resulting in substantial activation of CDK 9.
The term "cell proliferative disorder" includes disorders associated with adverse or uncontrolled proliferation of cells and it includes disorders associated with reduced or insufficient programmed cell death (apoptosis). The compounds of the invention are useful for preventing, inhibiting, blocking, reducing, controlling cell proliferation and/or cell division and/or inducing apoptosis, and the like. The present methods comprise administering to a subject (including mammals, including humans) in need thereof an amount of a compound of the present invention, or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate or solvate thereof, effective to treat or prevent the disorder.
Infectious diseases including opportunistic infections
In another aspect of the present invention, the compounds of formula (I) are used for the preparation of pharmaceutical compositions for the prevention and/or treatment of infectious diseases including opportunistic diseases and opportunistic infections. The term infectious disease includes infections caused by viruses, bacteria, prions, fungi and/or parasites.
Viral-induced infectious diseases including opportunistic diseases are specifically mentioned. In a preferred embodiment of this aspect, the virally-induced infectious disease, including an opportunistic disease, is caused by a retrovirus, Human Endogenous Retrovirus (HERV), hepadnavirus, herpesvirus, flavivirus and/or adenovirus. Preferably, the retrovirus is selected from a lentivirus or an oncogenic retrovirus, wherein the lentivirus is preferably selected from: HIV-1, HIV-2, Feline Immunodeficiency Virus (FIV), Bovine Immunodeficiency Virus (BIV), Simian Immunodeficiency Virus (SIV), chimeras of HIV and SIV (SHIV), Caprine Arthritis Encephalitis Virus (CAEV), ovine demyelinating encephalovitis/chronic progressive pulmonary disease virus (visna/maedivirus, VMV) or Equine Infectious Anemia Virus (EIAV), preferably HIV-1 and HIV-2, and the oncogenic retrovirus is preferably selected from HTLV-I, HTLV-II or Bovine Leukemia Virus (BLV), preferably HTLV-I and HTLV-II.
Said hepadnavirus is preferably selected from HBV, geotrichus hepatitis virus (GSHV) or Woodchuck Hepatitis Virus (WHV), preferably HBV, said herpesvirus being selected from: herpes Simplex Virus I (HSVI), herpes simplex virus II (HSVII), Epstein-Barr virus (EBV), Varicella Zoster Virus (VZV), Human Cytomegalovirus (HCMV) or human herpes virus type 8 (HHV-8), preferably HCMV, and the flavivirus is selected from HCV, West Nile virus or yellow fever virus.
It is understood that all of the above viruses also include resistant strains of the virus.
Examples of infectious diseases are aids, alveolar echinococcosis (AHD, echinococcosis), amebiasis (entamoeba histolytica infection), strongyloides vascularis infection, heteroscedasia, anthrax, babesiosis (babesia infection), parvus infection (venocytosis), bailey ascaris infection (ascaris), schizophragma disease (schistosomiasis), human yeast infection (blastomycosis), borreliosis, botulism, Brainerd diarrhea, brucellosis, BSE (spongiform encephalopathy), candidiasis, capillariosis (telangiitis infection), CFS (chronic fatigue syndrome), chagas disease (trypanosomiasis), chicken pox (varicella zoster virus), chlamydia pneumoniae infection, cholera, CJD (creutzfeldt-jakob disease), clonorchiasis sinensis infection (clonorchiasis), CLM (cutaneous larva immigration, hookworm infection), Coccidioidomycosis, conjunctivitis, coxsackie virus a16 (hand-foot-and-mouth disease), cryptococcosis, cryptosporidiosis infection (cryptosporidiosis), culex (vector of west nile virus), cyclosporinosis (cyclosporinosis infection), cysticerosis (cysticercosis of nervous system), cytomegalovirus infection, dengue/dengue fever, taenia dorsalis infection (flea canis and flea tapeworm), ebola hemorrhagic fever, echinococcosis (alveolar echinococcosis), encephalitis, intracolonic amoeba infection, dispar amoeba infection, amoeba harderi infection, histolytic amoeba infection (amebiasis), amoeba bornandii infection, enterobiasis infection (non-poliomyelitis), Epstein-Barr virus infection, escherichia coli infection, food-borne infection, foot and mouth disease, fungal dermatitis, gastroenteritis, streptococcus agalosis (streptococcus agalactis), streptococcus agalactiae (streptococcus mutans) infection, Streptococcal B disease (GroupBstrepcocculaldisease), Hanseng disease (leprosy), Hantaan virus pulmonary syndrome, head lice infestation (pediculosis), infection by helicobacter pylori, hematologic diseases, infection by Hendra virus, hepatitis (HCV, HBV), herpes zoster (Shingles)), HIV infection, human Ehrlichiosis, human parainfluenza infection, influenza, Isosporocysticerosis (isosporozococcidiosis infection), Lassa fever, leishmaniasis, kala fever (kala, Leishmania parasite), leprosy, lice (body lice, head lice, pubic lice), Lyme disease, malaria, Marburg hemorrhagic fever, measles, meningitis, mosquito borne disease, Mycobacterium Avium Complex (MAC) infection, Nadelbrueckia infection, nosocomiasis, nonpathogenic enteric amoeba infection, onchocerciasis (Ophiosis), liver trematosis (hepatitis), parvovirus (Ophiosis), parvovirus infection, Plague, PCP (pneumocystis pneumonia), poliomyelitis, Q fever, rabies, Respiratory Syncytial Virus (RSV) infection, rheumatic fever, rift valley fever, rotavirus infection, nematode infection, salmonella enteritidis, scabies, shigellasis, herpes zoster, sleep disorders, smallpox, streptococcal infection, taeniasis (taeniasis), tetanus, toxic shock syndrome, tuberculosis, ulcers (peptic ulcer disease), brook fever, vibrio parahaemolyticus infection, vibrio vulnificus infection, viral hemorrhagic fever, warts, waterborne infectious diseases, west nile virus infection (west nile encephalitis), pertussis, yellow fever.
Immune disease
Another aspect of the present invention relates to the use of at least one compound of general formula (I) and/or a pharmaceutically acceptable salt thereof for the prevention and/or treatment of immune, neuroimmune and autoimmune diseases.
Immunological diseases are, for example, asthma and diabetes, rheumatic and autoimmune diseases, aids, transplant organ and tissue rejection (as described below), rhinitis, chronic obstructive pulmonary disease, osteoporosis, ulcerative colitis, sinusitis, lupus erythematosus, recurrent infections, atopic dermatitis/eczema and occupational allergies, food allergies, drug allergies, severe allergies, allergic reactions and other manifestations of allergic diseases, as well as rare diseases such as primary immunodeficiency including antibody deficiency, cell-mediated immunodeficiency (e.g., severe combined immunodeficiency syndrome, DiGeorge syndrome, hyper-IgE syndrome, weil-alder syndrome, dyskinetic telangiectasia), immune-mediated cancer and leukocyte deficiency.
In autoimmune diseases, for example systemic lupus erythematosus, Rheumatoid Arthritis (RA), Multiple Sclerosis (MS), immune-mediated or type I diabetes, immune-mediated glomerulonephritis, scleroderma, pernicious anemia, alopecia, pemphigus vulgaris, myasthenia gravis, inflammatory bowel disease, crohn's disease, psoriasis, autoimmune thyroid disease, as well as hashimoto's disease, dermatomyositis, goodpasture's syndrome, pseudoparalytic myasthenia gravis, sympathetic ophthalmia, phakogeneuveitis (phakogeneuveitis), chronic active hepatitis, primary biliary cirrhosis, autoimmune hemolytic anemia, weichhoff's disease, uncontrolled attack of specific cells on the body's own tissues and organs (autoimmunity), development of inflammatory reactions and other serious symptoms and diseases.
Hashimoto's thyroiditis is one of the most common autoimmune diseases. "autoimmune disease" refers to a group of more than 80 chronic diseases, each of widely varying nature, which can affect organs such as the kidneys and any part of the digestive system from endocrine glands (e.g., thyroid).
There are many different autoimmune diseases and each can affect the human body in different ways. For example, autoimmune reactions are directed to the brain in multiple sclerosis and to the gut in crohn's disease. In other autoimmune diseases, such as systemic lupus erythematosus (lupus), diseased tissues and organs may vary between individuals with the same disease. One individual with lupus may have skin and joint damage, while another individual may have skin, kidney and lung damage. Finally, damage to certain tissues by the immune system may be permanent, as may damage to the insulin-producing cells of the pancreas in type I diabetes.
Cardiovascular diseases
The compounds of the invention are also useful in the prevention and/or treatment of cardiovascular diseases such as cardiac hypertrophy, adult congenital heart disease, aneurysm, stable angina, unstable angina, angioneurotic edema, aortic stenosis, aortic aneurysm, arrhythmia, arrhythmic right ventricular dysplasia, arteriosclerosis, arteriovenous malformations, atrial fibrillation, white plug syndrome, bradycardia, cardiac tamponade, cardiac dilation, congestive cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, cardiovascular disease prevention, carotid stenosis, cerebral hemorrhage, Churg-Strauss syndrome, diabetes, erbstein's abnormality (Ebstein's sAnomaly), eismenger's complex, cholesterol embolism, bacterial endocarditis, fibromyalgia, congenital heart defects, heart disease, congestive heart failure, valvular disease, cardiac infarction, stroke, heart attack, epidural hematoma, subdural hematoma, Hippel-Lindau disease, congestion, hypertension, pulmonary hypertension, hypertrophic growth, left ventricular hypertrophy, right ventricular hypertrophy, left cardiac dysplasia syndrome, hypotension, intermittent claudication, ischemic heart disease, crit-weitsie (Klippel-Trenaunay-Weber) syndrome, lateral bulbar syndrome, QT prolongation syndrome, mitral valve prolapse, smog disease, mucocutaneous lymph node syndrome, myocardial infarction, myocardial ischemia, myocarditis, pericarditis, peripheral vascular disease, phlebitis, polyarteritis nodosa, pulmonary valve occlusion, raynaud's disease, restenosis, Sneddon syndrome, stenosis, venous syndrome, syndrome X, tachycardia, takayasu's arteritis, hereditary hemorrhagic telangiectasia, temporal arteritis, farneso-tetranectasis, stroke syndrome, thromboangiitis, thrombosis, thromboembolism, tricuspid valve occlusion, varicose veins, vascular disease, vasculitis, vasospasm, ventricular fibrillation, Williams' syndrome, peripheral vascular disease, varicose veins and leg ulcers, deep vein thrombosis, Wolff-Parkinson-White syndrome.
Preferably cardiac hypertrophy, adult congenital heart disease, aneurysm, angina (angina), angina (anginapectoris), arrhythmia, cardiovascular disease prevention, cardiomyopathy, congestive heart failure, myocardial infarction, pulmonary hypertension, hypertrophic growth, restenosis, stenosis, thrombosis, and arteriosclerosis.
Proliferative diseases
In another preferred embodiment, the cell proliferative disease is cancer, which is preferably selected from the group consisting of:
the proliferative disorder and cancer are preferably selected from: adenocarcinoma, choroidal melanoma, acute leukemia, acoustic schwannoma, ampulla, anal cancer, astrocytoma, basal cell carcinoma, pancreatic cancer, desmoid tumor, bladder cancer, bronchial cancer, breast cancer, burkitt's lymphoma, uterine cancer, small intestine tumor, ovarian cancer, endometrial cancer, ependymoma, epithelial cancers, Ewing's tumor, gastrointestinal cancer, gastric cancer, gallbladder cancer (gallobladdercancer), gallbladder cancer (gallobladdercarcinosa), uterine cancer, cervical cancer, glioblastoma, gynecological tumor, otorhinolaryngologic tumor, hematological tumor, hairy cell leukemia, urinary tract cancer, skin testicular cancer (skintestinisccer), brain tumor (glioma), brain metastasis, testicular cancer, pituitary tumor, carcinoid tumor, western sarcoma, laryngeal cancer, blastoma, bone cancer, colorectal cancer, Head and neck tumors (tumors of the ear, nose and throat), colon cancer, craniopharyngeal tumor, oral cancer (cancers in the oral cavity and lips), central nervous system cancer, liver metastasis, leukemia, palpebral tumors, lung cancer, lymph node cancer (hodgkin lymphoma/non-hodgkin lymphoma), lymphoma, stomach cancer, malignant melanoma, malignant neoplasia, gastrointestinal malignant tumors, breast cancer, rectal cancer, medulloblastoma, melanoma, meningioma, hodgkin's disease, mycoses, nasal cancer, schwannoma, neuroblastoma, kidney cancer, renal cell carcinoma, oligodendroglioma, esophageal cancer, osteolytic and osteogenic cancers, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, plasmacytoma, prostate cancer, pharyngeal cancer, rectal cancer, retinoblastoma, vaginal cancer, thyroid cancer, Schneeberg disease, esophageal cancer, spinolioms, T-cell lymphoma (mycosis), Thymoma, ductal carcinoma (tubeccinoma), ocular tumor, urethral carcinoma, urinary tumor, urothelial carcinoma, vulvar carcinoma, wart (wart), soft tissue tumor, soft tissue sarcoma, Wilm tumor, cervical carcinoma, tongue carcinoma, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, lobular carcinoma in situ, small cell lung carcinoma, non-small cell lung carcinoma, bronchial adenoma, pleuropulmonary blastoma, mesothelioma, brain stem glioma, hypothalamic glioma (hypophosphatemicioma), cerebellar astrocytoma, neuroectodermal tumor, pinealoma, uterine sarcoma, salivary gland carcinoma, anal adenocarcinoma, mast cell tumor, pelvic tumor, ureteroma, hereditary papillary renal carcinoma, intraocular melanoma, hepatocellular carcinoma (bile duct carcinoma with or without fibrolamellar change (hepatorenal carcinoma), epithelial carcinoma (intrahepatic cholangiocarcinoma), epithelial carcinoma with or mixed bile duct carcinoma), biliary tract carcinoma (cholangiocarcinoma), bladder carcinoma, bladder, Squamous cell carcinoma, malignant melanoma, merkel cell skin carcinoma, non-melanoma skin carcinoma, hypopharynx cancer, nasopharyngeal cancer, oropharyngeal cancer, oral cancer, squamous cell carcinoma, oral melanoma, aids-related lymphoma, cutaneous T-cell lymphoma, central nervous system lymphoma, malignant fibrous histiocytoma, lymphosarcoma, rhabdomyosarcoma, malignant histiocytosis, fibrosarcoma, angiosarcoma, hemangiothecoma, leiomyosarcoma, canine breast cancer, and feline breast cancer.
The following cancer types are preferred: leukemias (including, but not limited to, chronic lymphocytic leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, mixed leukemia), bladder cancer, breast cancer (clearcancer), central nervous system cancer, colon cancer, stomach cancer, lung cancer, kidney cancer, melanoma, head and neck tumors (tumors of the ear, nose and throat), ovarian cancer (ovaricancancer), ovarian cancer (ovarialcarconema), cervical cancer (cervicalcencer), cervical cancer (cervicalcercinoma), glioblastoma, pancreatic cancer (pancreaticincanccer), pancreatic cancer (pancreaticincancorma), prostate cancer, stomach cancer, skin testicular cancer, hodgkin's lymphoma, liver cancer, liver metastases, and renal cell carcinoma.
Inflammation(s)
In another preferred embodiment, the inflammation is preferably mediated by the cytokines TNF- α, IL-1 β, GM-CSF, IL-6 and/or IL-8.
As mentioned above, the compounds of general formula (I) are pharmaceutically active substances for the prophylaxis and/or treatment of inflammatory diseases. Accordingly, these compounds are useful for the preparation of pharmaceutical formulations for the prevention and/or treatment of inflammation and inflammatory diseases in mammals, including humans.
Inflammatory diseases may result from infectious and non-infectious inflammatory conditions, which may be caused by an invasive organism or by an irritating, traumatic, metabolic, allergic, autoimmune or idiopathic cause as shown in the following list
I. Acute infection
A. Viral b. bacterial
Non-infectious cause
Chronic (granulomatous) disease
A. Bacterial b. helicity
C. Mycotic (fungal) d. idiopathic
Allergic, immunological and idiopathic disorders
A. Hypersensitivity reactions
B. Immunological and idiopathic disorders
Other inflammatory disorders
A. Parasitic infection
B. Reasons for inhalation: acute (thermal) injury
-pollution and inhalation allergy
-carcinogens
C, radiation damage: necrosis by radiation
Thus, the compounds disclosed herein are useful for preventing and/or treating inflammation caused by invasive organisms such as viruses, bacteria, prions and parasites, as well as for preventing and/or treating inflammation caused by irritant, traumatic, metabolic, allergic, autoimmune or idiopathic causes.
Accordingly, the compounds disclosed herein are useful for the prevention and/or treatment of inflammatory diseases caused or contributed to by viruses, parasites and bacteria associated with or associated with inflammation.
The following bacteria are known to cause inflammatory diseases: mycoplasma pneumoniae (causing e.g. Chronic Lung Disease (CLD), chronic respiratory diseases in mice), mycoplasma urealyticum (causing neonatal pneumonia), mycoplasma pneumoniae and chlamydia pneumoniae (causing chronic asthma), chlamydia pneumoniae (c.
The following viruses are known to cause inflammatory diseases: herpes viruses are in particular cytomegaloviruses (causing coronary heart disease in humans).
The compounds disclosed herein are useful for the prevention and/or treatment of inflammatory diseases caused and/or induced and/or caused and/or enhanced by the above mentioned bacteria or viruses.
In addition, the compounds of formula (I) are useful for the prevention and/or treatment of inflammatory diseases of the Central Nervous System (CNS), inflammatory rheumatic diseases, inflammatory diseases of the blood vessels, inflammatory diseases of the middle ear, inflammatory diseases of the intestinal tract, inflammatory diseases of the skin, inflammatory diseases of uveitis, inflammatory diseases of the larynx.
Examples of inflammatory diseases of the Central Nervous System (CNS) are algiosis, prototheca, bacterial disorders, abscesses, bacterial meningitis, idiopathic inflammatory disorders, eosinophilic meningoencephalitis, feline encephalomyelitis, granulomatous meningoencephalitis, meningitis, steroid-reactive meningitis-arteritis, other meningitis/meningoencephalitis, greyhous meningoencephalitis, necrotizing encephalitis, pyogenic granulomatous meningoencephalitis, dog wobble, mycotic diseases of the central nervous system, parasitic encephalomyelitis, prion protein-induced diseases, feline spongiform encephalopathy, protozoal encephalitis-myelitis, toxoplasmosis, neosporosis, sarcosporosis, encephalitozoonosis, trypanosomiasis, acanthamoebasis, babesiosis, leishmaniasis, rickettsia, rockborne mountain spotted fever, canine ehrlichiosis, bacterial meningitis, feline encephalomyelitis, necrobiosis, encephalitis-induced diseases, and encephalitis, Salmon meat poisoning, viral disorders, Aujeszky's disease, Borna disease, canine herpes virus viral encephalomyelitis, canine distemper encephalitis, canine distemper encephalomyelitis in immature animals, chronic relapsing encephalomyelitis, post-vaccination canine distemper encephalitis, feline immunodeficiency virus, feline infectious peritonitis, feline leukemia virus, canine infectious hepatitis, Lakras encephalitis virus, parvoviral encephalitis, rabies, post-vaccination rabies.
Examples of inflammatory rheumatic diseases are rheumatoid arthritis, scleroderma, lupus, polymyositis, dermatomyositis, psoriatic arthritis, ankylosing spondylitis, reiter's syndrome, juvenile rheumatoid arthritis, bursitis, tendonitis (tendonitis) and fibromyositis.
Examples of vascular inflammatory diseases are vasculitis, autoantibodies in vasculitis, microscopic polyangiitis, giant cell arteritis, multiple arteritis, vasculitis of the central nervous system, thromboangiitis obliterans (Buerger's disease), vasculitis secondary to bacterial, fungal and parasitic infections, vasculitis and rheumatoid arthritis, vasculitis in systemic lupus erythematosus, vasculitis in idiopathic inflammatory myopathy, relapsing polychondritis, systemic vasculitis in sarcoidosis, vasculitis and malignancy, and drug-induced vasculitis.
Examples of inflammatory diseases of the middle ear are acute suppurative otitis media, bullous myringitis, granulomatous myringitis and chronic suppurative otitis media, which may manifest as mucosis, cholesteatoma or both.
Examples of inflammatory bowel diseases are ulcerative colitis, crohn's disease.
Examples of inflammatory diseases of the skin are inflammatory dermatoses, urticaria (urticaria), edematous dermatitis of the spinous cell layer, allergic contact dermatitis, irritant contact dermatitis, atopic dermatitis, erythema multiforme (light EM), stewart-johnson syndrome (SJS, heavy EM), Toxic Epidermal Necrolysis (TEN), chronic inflammatory dermatoses, psoriasis, lichen planus, discoid lupus erythematosus and acne vulgaris.
Uveitis is an inflammation that is located in and/or on the eye and may be associated with inflammation elsewhere in the body. In most cases, patients with uveitis, which is part of a disease in other parts of the body, should be alerted to the disease. Most uveitis patients do not have clearly associated systemic disease. The etiology of uveitis can be infectious, camouflage syndrome (masqueradesyn), suspected immune-mediated diseases, and/or syndromes primarily confined to the eye.
The following viruses are associated with inflammation: human immunodeficiency virus-I, herpes simplex virus, herpes zoster virus and cytomegalovirus.
The inflammation caused, induced and/or enhanced by the bacteria or spirochetes is tuberculosis, leprosy, propionibacteria, syphilis, whipple's disease, leptospirosis, brucellosis and lyme disease.
The inflammation caused, induced, caused and/or enhanced by the parasite (protozoan or helminth) is toxoplasmosis, acanthamoeba disease, toxocariasis, cysticercosis, onchocerciasis.
Examples of inflammatory diseases caused, induced and/or enhanced by fungi are histoplasmosis, coccidioidomycosis, candidiasis, aspergillosis, sporotrichosis, blastomycosis and cryptococcosis.
Camouflage syndromes are, for example, leukemias, lymphomas, retinitis pigmentosa and retinoblastomas.
The suspected immune-mediated disease may be selected from: ankylosing spondylitis, Behcet's disease, Crohn's disease, drug reactions or hypersensitivity reactions, interstitial nephritis, juvenile rheumatoid arthritis, Kawasaki disease, multiple sclerosis, psoriatic arthritis, Ruite's syndrome, relapsing polychondritis, sarcoidosis, Sjogren's syndrome, systemic lupus erythematosus, ulcerative colitis, vasculitis, vitiligo, Vout-Taurus-Tahitian syndrome.
Syndromes primarily confined to the eye are, for example, acute multifocal squamous pigment epithelium, acute retinal necrosis, shotgun bullet-like choroidopathies, Fuchs' heterochromitic cyclitis, glaucomatous cyclitis crisis, lens-induced uveitis, multifocal choroiditis, pars plana cyclitis, creeping choroiditis, sympathetic ophthalmia and trauma.
Examples of inflammatory diseases of the larynx are gastroesophageal (laryngo) reflux disease, children's laryngitis, adult tight larynx, chronic (granulomatous) diseases, allergic, immunological and idiopathic disorders, and other inflammatory conditions.
Laryngitis in children is known as an acute (viral or bacterial) infection, such as laryngotracheitis (croup), supraglottitis (epiglottitis), diphtheria, non-infectious diseases due to, for example, wheezy laryngospasm and traumatic laryngitis.
Adult laryngopharyngeal procedures are, for example, viral laryngitis, common upper respiratory tract infections, laryngotracheitis, herpes simplex, bacterial laryngitis, supraglottitis, laryngeal abscesses, and gonorrhea.
The chronic (granulomatous) disease may be selected from bacterial diseases, tuberculosis, leprosy, scleroma, actinomycosis, tularemia, melioidosis, spirochaetes (syphilis) disease, mycotic (fungal) disease, candidiasis, blastomycosis, histoplasmosis, coccidioidomycosis, aspergillosis, idiopathic diseases, sarcoidosis and wegener's granulomatosis.
Allergic, immunological and idiopathic conditions are, for example, hypersensitivity reactions, angioedema, Stevens-Johnson syndrome, immunological and idiopathic conditions, infections of immunocompromised hosts, rheumatoid arthritis, systemic lupus erythematosus, cicatricial pemphigoid, recurrent polychondritis, Sjogren's syndrome and amyloidosis.
Other inflammatory conditions are, for example, parasitic infections, trichinosis, leishmaniasis, schistosomiasis, praecox laryngis, inhaled laryngitis, acute (thermal) injury, pollution and inhaled allergy, carcinogens, radiation injury, radiation laryngitis, radiation necrosis, vocal cord bleeding, dystonia, and contact ulcers and granulomas.
Apoplexy (apoplexy)
The compounds of general formula (I) and their pharmaceutically acceptable salts of the present invention are also useful in the treatment of stroke.
In another aspect of the invention, the compounds of formula (I) and pharmaceutically acceptable salts thereof are useful as inhibitors of protein kinases, preferably cellular protein kinases.
In a preferred embodiment of this aspect, the cellular protein kinase comprises a Cyclin Dependent Kinase (CDK).
The cyclin-dependent kinase may be selected from the group consisting of: CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, crks (Crk7, CDC 2-related protein kinase 7), CDKL1 (cyclin-dependent kinase-like 1), KKIALRE, CDKL2 (cyclin-dependent kinase-like 2), KKIAMRE, CDKL3 (cyclin-dependent kinase-like 3), NKIAMRE, CDKL4 similar to cyclin-dependent kinase-like 1, CDC2L1 (cell-division cycle 2-like 1), pieb, CDC2L1 (cell-division cycle 2-like 1), pittslrea, CDC2L5 (cell-division cycle 2-like 5), PCTK1 (taptk protein kinase 1), pctatk 2 (pcire protein kinase 2), pfirek 3 (pcirer 1) or taptk 1 (cyclin-dependent kinase-like 3).
In a particularly preferred embodiment, the cyclin-dependent kinase is CDK 9. The compounds of general formula (I) and their pharmaceutically acceptable salts are therefore useful as inhibitors of CDK 9.
In addition, in another particularly preferred embodiment, the compounds of the invention show high efficacy (from low IC) in inhibiting CDK9 activity50Proof of value). In thatIn the context of the present invention, the IC to CDK9 may be determined by the method described in the methods section below50The value is obtained. Preferably, it is determined according to the method described in section 3.6.
Surprisingly, the results show that the compounds of general formula (I) and their pharmaceutically acceptable salts selectively inhibit CDK9, but not other protein kinases or other cyclin-dependent kinases. Accordingly, the compounds of general formula (I) and their pharmaceutically acceptable salts are useful as selective inhibitors of CDK 9.
Particularly preferred compounds of formula (I) according to the invention exhibit a greater inhibition of CDK9 than CDK 2. In the context of the present invention, the IC to CDK2 may be determined by the method described in the methods section below50The value is obtained. Preferably, it is determined according to the method described in section 3.5.
In addition, the compounds of formula (I) according to the invention mediate an antiproliferative activity in tumor cell lines such as HeLa, MaTu/ADR, H460, DU145, CACO-2 or B16F 10. In the context of the present invention, the IC of the compounds on these cell lines is preferably determined according to the method described below50The value is obtained.
Preferred compounds of the invention mediate a particularly strong antiproliferative activity in the tumour cell line HeLa.
As used herein, a kinase "inhibitor" refers to any compound capable of downregulating, reducing, inhibiting, or modulating the amount/activity of a kinase. Inhibition of these kinases can be achieved by any of a variety of mechanisms known in the art, including but not limited to direct binding to the kinase polypeptide, denaturing or inactivating the kinase, or inhibiting expression of a gene encoding the kinase (e.g., transcription into mRNA, translation into nascent polypeptide, and/or final polypeptide modification into mature protein). In general, kinase inhibitors may be proteins, polypeptides, nucleic acids, small molecules, or other chemical groups.
The term "inhibit" as used herein refers to the ability of a compound to at least partially down-regulate, reduce, neutralize, inactivate or inhibit an enzyme activity or the expression of an enzyme or protein and/or viral replication.
In another aspect of the present invention, there is provided a method for the prophylaxis and/or treatment of infectious diseases including opportunistic diseases in a mammal, particularly a human, comprising administering to the mammal an amount of at least one compound of formula (I) effective in the prophylaxis and/or treatment of said infectious diseases including opportunistic diseases. In a preferred embodiment of the method, the infectious disease, including opportunistic diseases, is a virus-induced infectious disease. The virus-induced infectious disease including opportunistic diseases is caused by retroviruses, hepadnaviruses, herpesviruses, flaviviruses and/or adenoviruses. In another preferred embodiment of the method, the retrovirus is selected from a lentivirus or an oncoretrovirus, wherein the lentivirus is selected from the group consisting of: HIV-1, HIV-2, FIV, BIV, SIV, SHIV, CAEV, VMV or EIAV, preferably HIV-1 or HIV-2, and wherein the oncogenic retrovirus is selected from HTLV-I, HTLV-II or BLV. In another preferred embodiment of the method, said hepadnavirus is selected from HBV, GSHV or WHV, preferably HBV, said herpesvirus is selected from: HSVI, HSVII, EBV, VZV, HCMV or HHV8, preferably HCMV, and the flavivirus is selected from HCV, West Nile virus and yellow fever virus.
In another aspect of the present invention, there is provided a method for preventing and/or treating infectious diseases including opportunistic diseases, prion diseases, immunological diseases, autoimmune diseases, cardiovascular diseases, cell proliferative diseases, inflammation, erectile dysfunction and stroke in a mammal, particularly a human, comprising administering to said mammal an amount of at least one compound of the general formula (I) and/or a pharmaceutically acceptable salt thereof effective to prevent and/or treat said infectious diseases including opportunistic diseases, prion diseases, immunological diseases, autoimmune diseases, cardiovascular diseases, cell proliferative diseases, inflammation, erectile dysfunction and stroke.
In another preferred embodiment, the specific diseases referred to as infectious diseases including opportunistic diseases, prion diseases, immunological diseases, autoimmune diseases, cardiovascular diseases, cell proliferative diseases, inflammation, erectile dysfunction and stroke are selected from the group disclosed above.
The compounds explicitly shown in table 1 are preferably used in the methods and indications disclosed herein. Another aspect of the invention is the administration of at least one compound of the general formula (I) for use as a pharmaceutically active substance in combination with another therapeutic compound.
For the indication HIV, compounds of general formula (I), preferably those shown in table 4 for CDK9, may be administered in combination with antiretroviral drugs selected from the following five classes:
1) nucleoside Reverse Transcriptase Inhibitors (NRTI),
2) Non-nucleoside reverse transcriptase inhibitors (NNRTIs),
3) Protease Inhibitors (PI),
4) A fusion inhibitor, or
5) An immunostimulant.
Pharmaceutical composition and pharmaceutical combination
Another aspect of the present invention relates to pharmaceutical combinations and pharmaceutical compositions comprising as active ingredient at least one compound of general formula (I) and at least one pharmaceutically acceptable carrier, excipient and/or diluent and optionally one or more other anti-neoplastic agents or one or more anti-retroviral agents. The term "pharmaceutical combination" as used herein refers to a combination of at least two pharmaceutically active substances or therapeutic agents, with or without additional ingredients, carriers, diluents and/or solvents. The term "pharmaceutical composition" as used herein refers to a galenical formulation of at least one pharmaceutically active substance with at least one further ingredient, carrier, diluent and/or solvent.
The compounds of formula (I) may be administered as the sole agent or in combination with one or more additional therapeutic agents, wherein the pharmaceutical combination does not cause unacceptable adverse effects. The combination therapy includes administration of a single pharmaceutical dosage formulation comprising a compound of formula (I) and one or more additional therapeutic agents as separate pharmaceutical compositions, as well as administration of a compound of formula (I) and each additional therapeutic agent in separate pharmaceutical dosage formulations (i.e., in separate pharmaceutical compositions). For example, the compound of formula (I) and the therapeutic agent may be administered to the patient together in a single oral dosage composition, e.g., in the form of a tablet or capsule, or each agent may be administered as separate pharmaceutical compositions.
When separate pharmaceutical compositions are used, the compound of formula (I) and one or more additional therapeutic agents may be administered at substantially the same time (e.g., simultaneously) or at staggered times from one another (e.g., sequentially).
In particular, the compounds of the present invention may be used in fixed (fixed) or separate pharmaceutical compositions containing other antineoplastic agents, such as alkylating agents, antimetabolites, plant-derived antineoplastic agents, hormonal therapy agents, topoisomerase inhibitors, camptothecin derivatives, kinase inhibitors, targeting drugs, antibodies, interferons and/or biological response modifiers, anti-angiogenic compounds and other antineoplastic agents. In this regard, the following is a non-limiting list of second agents that may be used in combination with the compounds of the present invention:
● alkylating agents including, but not limited to, nitrogen mustards, N-oxides, cyclophosphamide, ifosfamide, thiotepa, ranimustine, nimustine, temozolomide, altretamine, apaziquone, bromtalin, bendamustine, carmustine, estramustine, fotemustine, glufosfamide, cyclophosphamide and dibromodulcitol; platinum coordination alkylating compounds including, but not limited to, cisplatin, carboplatin, eptaplatin, lobaplatin, nedaplatin, oxaliplatin and satraplatin;
● antimetabolites including, but not limited to, methotrexate, 6-mercaptopurine, inosine, mercaptopurine, 5-fluorouracil, tegafur, doxifluridine, carmofur, cytarabine octadecylphosphate (cytarabine ocfosfate), enocitabine, gemcitabine, fludarabine, 5-azacitidine, capecitabine, cladribine, clofarabine, decitabine, efluorine, ethynylcytidine (ethylcytidine), cytarabine, hydroxyurea, melphalan, nelarabine, nolatrexed, ocfosfilte, disodium pemetrexed, pellitorine, raltrexed, tripaine, trimetrexate, vidarabine, vincristine, and vinorelbine, alone or in combination with folinic acid;
● hormonal therapy agents including, but not limited to, exemestane, leuprolide acetate, anastrozole, doxercalciferol, fadrozole, formestane, 11-beta hydroxysteroid dehydrogenase 1 inhibitors, 17-alpha hydroxylase/17, 20 lyase inhibitors such as abiraterone acetate, 5-alpha reductase inhibitors such as finasteride and epristeride, antiestrogens such as tamoxifen citrate and fulvestrant, triptorelin, toremifene, raloxifene, lasofoxifene, letrozole, antiestrogens such as bicalutamide, flutamide, mifepristone, nilutamide, Casodex, and antiprogestins, and combinations thereof;
● plant derived antitumor substances including, for example, those selected from mitotic inhibitors, such as epothilones, for example, sargapirone, ixabepilone and epothilone B, vinblastine, vinflunine, docetaxel and paclitaxel;
● cytotoxic topoisomerase inhibitors including, but not limited to, aclarubicin, doxorubicin, amonafide, piroxicam, camptothecin, 10-hydroxycamptothecin, 9-aminocamptothecin, diflucan, irinotecan, topotecan, eltrocoline, epirubicin, etoposide, exatecan, gimatecan, lurtotecan, mitoxantrone, pirarubicin, picotron, rubitecan, sobuzolz, tafluoroporphin, and combinations thereof;
● immunological agents including interferons such as interferon alpha, interferon alpha-2 a, interferon alpha-2 b, interferon beta, interferon gamma-1 a and interferon gamma-n 1, and other immunopotentiators such as L19-IL2 and other IL2 derivatives, filgrastim, lentinan, schizophyllan, TheraCys, ubenimex, aldesleukin, alemtuzumab, BAM-002, dacarbazine, dallizumab, dinil interleukin, gimumab, ozomicin, tiitumomab (ibritumomab), imiquimod, legumine, lentinan, melanoma vaccine (Corixa), moraxetin, sargrastim, thazinamine, tesicukin (tecleukin), thymalfasin (thymolasin), tositumomab, Vimlizin, epratuzumab, mitumumab, ovuzumab, panitumumab (pemumumab), and Provenge, Merial melanoma vaccines;
● bioresponse modifiers are agents which modulate the defense mechanisms or bioresponse of an organism such as the survival, growth or differentiation of tissue cells, thereby rendering them active against tumors; such agents include, for example, coriolus versicolor polysaccharide, lentinan, cezopyran, bisinby, promone, and ubenimex;
● anti-angiogenic compounds including, but not limited to, Avermectin, aflibercept, angiostatin, aplidine, arentar, acitinib, recentin, bevacizumab, alanine brimonib, cilengitide, combretastatin, DAST, endostatin, fenretinide, halofuginone, pazopanib, ranibizumab, removab, lenalidomide (revlimmid), sorafenib, vatalanib, squalamine, sunitinib, tiratinib, thalidomide, ukrain and vitaxin;
● antibodies including, but not limited to, trastuzumab, cetuximab, bevacizumab, rituximab, tositumomab (ticilimumab), ipilimumab, lutiximab, cetuximab, asexup, ogovazumab, and alemtuzumab;
● VEGF inhibitors, such as sorafenib, DAST, bevacizumab, sunitinib, recentin, axitinib, aflibercept, tiratinib, alanine brimonib, vatalanib, pazopanib nucleus and ranibizumab, Palladia;
● EGFR (HER1) inhibitors such as cetuximab, parlimumab, vectibix, gefitinib, erlotinib and Zactima;
● HER2 inhibitors such as lapatinib, trastuzumab and pertuzumab;
● mTOR inhibitors such as temsirolimus, sirolimus/rapamycin, and everolimus;
● c-Met inhibitors;
● P13K and AKT inhibitors;
● CDK inhibitors such as roscovitine and frataxiol;
● spindle assembly checkpoint inhibitors and targeted antimitotic agents, such as PLK inhibitors, Aurora inhibitors (e.g. hesperidin), checkpoint kinase inhibitors and KSP inhibitors;
● HDAC inhibitors such as panobinostat, Volinostat, MS275, Berlinostat, and LBH 589;
● HSP90 and HSP70 inhibitors;
● proteasome inhibitors such as bortezomib and carfilzomib;
● serine/threonine kinase inhibitors, including MEK inhibitors (e.g., RDEA119) and Raf inhibitors such as sorafenib;
● farnesyltransferase inhibitors, such as tipifarnib;
● tyrosine kinase inhibitors including, for example, dasatinib, nilotinib, DAST, bosutinib, sorafenib, bevacizumab, sunitinib, AZD2171, axitinib, aflibercept, tiratinib, imatinib mesylate, alanatinib, pazopanib, ranibizumab, vatalanib, cetuximab, panitumumab, vectibix, gefitinib, erlotinib, lapatinib, trastuzumab, pertuzumab and c-Kit inhibitors,
Palladia, masitinib;
● a vitamin D receptor agonist;
● Bcl-2 protein inhibitors such as olbaccarat, sodium orlimerson (oblimersensodium) and gossypol;
● cluster of differentiation 20 receptor antagonists, such as rituximab;
● ribonucleotide reductase inhibitors, such as gemcitabine;
● tumor necrosis apoptosis-inducing ligand receptor 1 agonists, such as mappaucimumab;
● 5-serotonin receptor antagonists, such as rEV598, zalonosetron, palonosetron hydrochloride, granisetron, Zindol and AB-1001;
● integrin inhibitors, including α 5- β 1 integrin inhibitors, such as E7820, JSM6425, voroximab, and endostatin;
● androgen receptor antagonists including, for example, nandrolone decanoate, fluoxymesterone, methyltestosterone, propadine (best-aid), andromostine, bicalutamide, flutamide, apo-cyproterone, apo-flutamide, chlormadinone acetate, andrurr, Tabi, cyproterone acetate, and nilutamide;
● aromatase inhibitors, such as anastrozole, letrozole, testolactone, exemestane, aminoglutethimide and formestane;
● matrix metalloproteinase inhibitors;
● other anticancer agents include, for example, alitretinoin, polyinosinic acid, atrasentan, bexarotene, bortezomib, bosentan, calcitriol, etosuline, fotemustine, ibandronic acid, miltefosine, mitoxantrone, I-asparaginase, procarbazine, dacarbazine, hydroxyurea, pemetrexed, pentostatin, tazarotene, bortezomib, gallium nitrate, camosfamide, darunavine, and tretinoin.
The compounds of the invention may also be used in the treatment of cancer in combination with radiotherapy and/or surgical intervention.
In addition, the compounds of formula (I) may be used alone or in compositions for research and diagnostics, or as analytical reference standards and the like, as are well known in the art.
Thus, a further aspect of the present invention relates to a pharmaceutical combination comprising at least one compound of general formula (I) according to the present invention and/or a pharmaceutically acceptable salt thereof and at least one antiretroviral drug, in particular at least one of the drugs mentioned above.
The pharmaceutical compositions of the present invention comprise at least one compound of the present invention as an active ingredient and at least one pharmaceutically acceptable (i.e., non-toxic) carrier, excipient and/or diluent. The pharmaceutical compositions of the present invention may be prepared in accordance with known procedures at suitable dosage levels in conventional solid or liquid carriers or diluents and conventional pharmaceutical adjuvants. Preferred formulations are suitable for oral administration. These administration forms include, for example, pills, tablets, film-coated tablets, capsules, powders and depots (deposits).
In addition, the present invention also encompasses pharmaceutical formulations for parenteral administration, including cutaneous, intradermal, intragastric, intradermal, intravascular, intravenous, intramuscular, intraperitoneal, intranasal, intravaginal, intraoral, transdermal, rectal, subcutaneous, sublingual, topical or transdermal (transdermal) administration, comprising at least one compound of the invention and/or a pharmaceutically acceptable salt thereof as active ingredient in addition to typical vehicles and/or diluents.
The pharmaceutical compositions of the present invention containing as active ingredient at least one compound of the present invention and/or pharmaceutically acceptable salts thereof will generally be administered together with suitable carrier materials selected with respect to the intended form of administration, i.e., for oral administration in the form of tablets, capsules (solid filled, semi-solid filled or liquid filled), powders for formulation, gels, elixirs, dispersible granules, syrups, suspensions and the like, and in accordance with conventional pharmaceutical practice. For example, for oral administration in the form of tablets or capsules, the active pharmaceutical ingredient may be combined with any oral, non-toxic pharmaceutically acceptable carrier, preferably an inert carrier, such as lactose, starch, sucrose, cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, talc, mannitol, ethanol (liquid filled capsules) and the like. In addition, suitable binders, lubricants, disintegrating agents and coloring agents may also be incorporated into the tablets or capsules. Powders and tablets may contain as active ingredient from about 5% to about 95% by weight of a4, 6-disubstituted pyrimidine derivative of general formula (I) or an analogous compound thereof or a pharmaceutically active salt of each.
Suitable binders include starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethylcellulose, polyethylene glycol and waxes. Suitable lubricants which may be mentioned are boric acid, sodium benzoate, sodium acetate, sodium chloride, etc. Suitable disintegrants include starch, methylcellulose, guar gum and the like. Sweetening and flavoring agents and preservatives may also be included as appropriate. The disintegrants, diluents, lubricants, binders, etc. are discussed in more detail below.
In addition, the pharmaceutical compositions of the present invention may be formulated in a sustained release form to provide a controlled rate of release of any one or more of the components or active ingredients to optimize the therapeutic effect, e.g., anti-histamine activity, and the like. Suitable dosage forms for sustained release include tablets having multiple layers with different rates of disintegration or a controlled release polymer matrix impregnated with the active ingredient and formed into a tablet shape, or capsules comprising such impregnated or encapsulated porous polymer matrices.
Liquid form preparations include solutions, suspensions, and emulsions. Examples which may be mentioned are aqueous or water/propylene glycol solutions for parenteral injection or solutions, suspensions and emulsions for oral administration with the addition of sweeteners and opacifiers. Liquid form preparations may also include solutions for intranasal administration.
Aerosols suitable for inhalation may include solutions and solids in powder form, which may be provided in combination with a pharmaceutically acceptable carrier, e.g., an inert compressed gas such as nitrogen.
To prepare suppositories, a low melting wax, e.g., a mixture of fatty acid glycerides, e.g., cocoa butter, is first melted and the active ingredient is then dispersed homogeneously therein, e.g., by stirring. The molten, homogeneous mixture is then poured into a suitably sized mould, allowed to cool and thus solidify.
Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral or parenteral administration. Such liquid form preparations include solutions, suspensions and emulsions.
The compounds of the present invention may also be delivered transdermally. The transdermal composition may have the form of a cream, lotion, aerosol and/or emulsion and may be contained in a matrix-type or depot-type transdermal patch known in the art for this purpose.
The term "capsule" as used herein refers to a specific container or shell made of methylcellulose, polyvinyl alcohol or modified gelatin or starch for encapsulating or containing a composition comprising one or more of the active ingredients. Capsules having a hard shell are typically made from a mixture of gelatin from bone or pig skin, which has a relatively high gel strength. The capsules themselves may contain small amounts of dyes, opacifiers, plasticizers and/or preservatives.
A tablet is understood to be a compressed or molded solid formulation comprising an active ingredient and a suitable diluent. Tablets may be prepared by compressing a mixture or granules obtained by wet granulation, dry granulation or compaction (compaction) methods well known to those of ordinary skill in the art.
Oral gels refer to active ingredients dispersed or dissolved in a hydrophilic semisolid matrix.
Powders for formulation refer to powder mixtures comprising the active ingredient and a suitable diluent, which may be suspended in, for example, water or fruit juice.
Suitable diluents are the substances which normally make up the major part of the composition or dosage form. Suitable diluents include sugars such as lactose, sucrose, mannitol and sorbitol, starches derived from wheat, corn, rice and potato, and celluloses such as microcrystalline cellulose. The amount of diluent in the composition may range from about 5% to about 95%, preferably from about 25% to about 75%, and more preferably from about 30% to about 60% by weight of the total weight of the composition.
The term disintegrant refers to a substance added to a composition to aid in breaking up (disintegrating) and releasing the pharmaceutically active ingredient of the drug. Suitable disintegrants include starches, "cold water soluble" modified starches such as sodium carboxymethyl starch, natural and synthetic gums such as locust bean gum, karaya gum, guar gum, tragacanth and agar, cellulose derivatives such as methylcellulose and sodium carboxymethyl cellulose, microcrystalline cellulose and cross-linked microcrystalline cellulose such as cross-linked sodium carboxymethyl cellulose, alginates such as alginic acid and sodium alginate, clays such as bentonite and effervescent mixtures. The amount of disintegrant in the composition may range from about 2% to about 20%, more preferably from about 5% to 10% by weight of the composition.
Binders are substances that bind or "glue" powder particles and make them sticky by forming particles, thus acting as "glue" in the formulation. The binder increases the existing cohesion of the diluent or filler. Suitable binders include sugars such as sucrose, starch from wheat, corn, rice and potato, natural and synthetic gums such as acacia, gelatin and tragacanth, seaweed derivatives such as alginic acid, sodium alginate and ammonium calcium alginate (ammoniumcalcaliginate), cellulosic materials such as methyl cellulose, sodium carboxymethyl cellulose and hypromellose, polyvinylpyrrolidone and inorganic compounds such as magnesium aluminum silicate. The amount of binder in the composition may range from about 2% to about 20%, preferably from about 3% to about 10%, and more preferably from about 3% to about 6% by weight of the composition.
Lubricants refer to a class of materials that are added to dosage forms to enable tablet particles and the like to be ejected from a die or mold after being compressed by reducing friction or wear. Suitable lubricants include metal stearates, such as magnesium, calcium or potassium stearate, stearic acid, high melting waxes and other water soluble lubricants such as sodium chloride, sodium benzoate, sodium acetate, sodium oleate, polyethylene glycol and D, L-leucine. Since lubricants must be present on the surface of the granules, they are usually added in the last step before tabletting. The amount of lubricant in the composition may range from about 0.2% to about 5% by weight of the composition, preferably from about 0.5% to about 2% by weight, and more preferably from about 0.3% to about 1.5% by weight of the composition.
Glidants are substances that prevent the components of a pharmaceutical composition from agglomerating and improve the flow characteristics of the granules, thereby making the flow smooth and uniform. Suitable glidants include silicon dioxide and talc. The amount of glidant in the composition may range from about 0.1 to about 5 weight percent, preferably from about 0.5 to about 2 weight percent of the final composition.
Colorants are excipients that color the composition or dosage form. Such excipients include food grade dyes adsorbed on a suitable adsorbent such as clay or alumina. The amount of colorant may range from about 0.1% to about 5%, preferably from about 0.1% to about 1% by weight of the composition.
Another aspect of the invention relates to a method of treatment. The method comprises administering a therapeutically effective amount of at least one compound of formula (I) as defined in claim 1 to a patient suffering from an infectious disease including an opportunistic disease, an immune disease, an autoimmune disease, a cardiovascular disease, a cell proliferative disease, an inflammation, an erectile dysfunction or a stroke, thereby treating the infectious disease including an opportunistic disease, an immune disease, an autoimmune disease, a cardiovascular disease, a cell proliferative disease, an inflammation, an erectile dysfunction or a stroke.
Another aspect of the present invention relates to a method comprising administering to a patient suffering from cancer, a tumor or a hyperproliferative disease a therapeutically effective amount of at least one compound of general formula (I) as defined in claim 1, thereby treating said cancer, tumor or hyperproliferative disease.
The invention is particularly useful for preventing the formation of metastases. In addition, the compounds of the present invention are particularly useful in the treatment of drug resistant and multi-drug resistant cancers and tumors.
Examples
Preparation of the Compounds
Abbreviations used in the chemical description and examples are as follows:
CDCl3(deuterated chloroform); cHex (cyclohexane); DCM (dichloromethane); DIPEA (diisopropylethylamine); DMF (dimethylformamide); DMSO (dimethyl sulfoxide); eq (equivalent); ES (electrospray); EtOAc (ethyl acetate); EtOH (ethanol); iPrOH (isopropanol); MeOH (methanol); MS (mass spectrometry); NMR (nuclear magnetic resonance); pd (dppf) Cl2(Dichloromethane-complexed [1, 1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride); iPrOH (isopropanol); RT (room temperature); sat. SiO 22(silica gel); TFA (trifluoroacetic acid); THF (tetrahydrofuran).
Preparation example
Intermediates
Intermediate 1: 3- [ (4-chloro-1, 3, 5-triazin-2-yl) amino group]Benzomethanesulfonamide (A1)
At 0 ℃ in N2To a solution of 2, 4-dichloro-1, 3, 5-triazine (1.0eq) in anhydrous DMF (0.7M) was added a solution of (3-aminophenyl) methanesulfonamide (1.0eq) in anhydrous DMF (0.7M) under an atmosphere. The reaction mixture was stirred at 0 ℃ for 2.5 h. Then water was added and saturated NaHCO was used3Neutralizing with water solution. The aqueous layer was extracted with EtOAc and Na2SO4The combined organic layers were dried. The solvent was evaporated to give a1 as a white solid, which was used in the next step without further purification.1HNMR(400MHz,d6-DMSO,300K)4.26(s,2H),6.89(s,2H),7.15(d,J=7.4Hz,1H),7.36(d,J=7.4Hz,1H),7.66(d,J=7.4Hz,1H),8.64(s,1H),9.75(br.s,1H),10.83(s,1H)。MS(ES)C10H10ClN5O2Theory of SThe value: 299, measured values: 300(M + H)+。
Intermediate 2: 3- [ (4-chloro-1, 3, 5-triazin-2-yl) amino group]Benzenesulfonamide (A2)
A2 was prepared according to the general procedure reported for a1, using 2, 4-dichloro-1, 3, 5-triazine and 3-aminobenzenesulfonamide as reaction materials. The crude product was purified by flash chromatography on silica gel (cehex/EtOAc =20:1-1:20) to give the desired product a2(35%) as a white solid.1HNMR(400MHz,d6-DMSO,300K)7.41(s,2H),7.59(m,2H),7.85(d,J=6.9Hz,1H),8.21(br.s,1H),8.69(s,1H),11.02(s,1H)。MS(ES)C9H8ClN5O2Theoretical value of S: 285, measured value: 286(M + H)+。
Intermediates3: 4-chloro-N- (3-nitrophenyl) -135-triazin-2-amine (A3)
A3 was prepared following the general procedure reported for A1, using 2, 4-dichloro-1, 3, 5-triazine and 3-nitroaniline as the reaction materials.1H-NMR(400MHz,d6-DMSO,300K)7.67(t,J=8.1Hz,1H),7.98(dd,J=8.1Hz,J=2.2Hz,1H),8.04(dd,J=8.1Hz,J=2.2Hz,1H),8.69(m,1H),8.77(m,1H),11.17(s,1H)。MS(ES)C9H6ClN5O2Theoretical value: 251, measurement value: 252(M + H)+。
Intermediate 4: 2- (3-aminophenyl) ethanesulfonamide (A4)
Compound a4 was obtained by reduction of 2- (3-nitrophenyl) ethanesulfonamide according to the procedure described in WO2009/076140, and according to j.med.chem.45(2002) 567-583 the nitro compound is obtained from 2- (3-nitrophenyl) ethanol.
Intermediate 5: 2- [3- ((4-chloro-1, 3, 5-triazin-2-yl) amino) phenyl]Ethanesulfonamide (A5)
A5 was prepared according to the general procedure reported for a1, using 2, 4-dichloro-1, 3, 5-triazine and 2- (3-aminophenyl) ethanesulfonamide a4 as reaction materials. The crude product was purified by flash chromatography on silica gel (DCM/MeOH =100:0-4:1) to afford as a crude productDesired product a5 (purity 54%, 65%) as a brown solid. MS (ES) C11H12ClN5O2Theoretical value of S: 313, measurement value: 314(M + H)+。
Intermediate 6: 3- [ (4-chloro-1, 3, 5-triazin-2-yl) amino group]Benzamide (A6)
A6 was prepared according to the general procedure reported for a1, using 2, 4-dichloro-1, 3, 5-triazine and 3-aminobenzamide as reaction materials. MS (ES) C10H8ClN5Theoretical value of O: 249, measurement: 250(M + H)+。
Intermediate 7: rac-S- [3- ((4-chloro-1, 3, 5-triazin-2-yl) amino) phenyl]-N-ethoxycarbonyl-S-methylsulfoxylimine (A7)
A solution of sulfoximine A11(500mg,2.064mmol) in a mixture of iPrOH (12ml) and THF (12ml) was cooled to-20 ℃. At this temperature, a further solution of pre-cooled 2, 4-dichloro-1, 3, 5-triazine (309.5mg,2.064mmol) in the same solvent mixture (6ml each) was added. After stirring for 1 hour, another batch of triazine (100mg,0.67mmol) was added and stirring continued at-20 ℃ for 1.5 hours. With saturated NaHCO3The mixture was adjusted to pH7 with aqueous solution and extracted with EtOAc. With Na2SO4The organic layer was dried and concentrated under reduced pressure to give product a7 as a yellow solid. Yield: 561.1mg (76%); MS (ES) C13H14ClN5O3Theoretical value of S: 355, measured value: 356(M + H)+。
Intermediate 8: 6- [ (4-chloro-1, 3, 5-triazin-2-yl) amino group]-2, 3-dihydro-1H-indole-1-sulfonamide (A8)
6-amino-2, 3-dihydro-1H-indole-1-sulfonamide hydrochloride was purchased from UkrOrg Synthesis and was used to prepare the title compound by reaction with 2, 4-dichloro-1, 3, 5-triazine as described for A7. Product A8 was obtained as a brown solid. MS (ES) C11H11ClN6O2Theoretical value of S: 326, measurement value: 327(M + H)+。
Intermediates9: 2- [2- ((4-pyridinyl) methoxy) phenyl]-4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan (A9)
To a solution of 2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenol (400mg,1.82mmol) in DMF (6ml) were added 4-chloromethylpyridine hydrochloride (4-picolichlorohydrochloride) (446mg,2.72mmol) and K2CO3(1.0g,7.24 mmol). The mixture was heated in a microwave oven at 150 ℃ for 1 hour. After addition of water and EtOAc, the organic layer was separated and washed with Na2SO4Dried and the solvent removed under reduced pressure. After chromatographic purification (silica gel, DCM/MeOH gradient 100:0-90:10), the title compound A9 was isolated as a white solid (15%). MS (ES) C18H22BNO3Theoretical value: 311, measurement value: 312(M + H)+。
Intermediate 10: 2- [2- (4- (tert-Butoxycarbonylamino) butoxy) phenyl]-4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan (A10)
The title compound a10 was prepared from 2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenol and tert-butyl N- (4-bromobutyl) carbamate in substantially the same manner as described for a 9. MS (ES) C21H34BNO5Theoretical value: 391, measurement value: 392(M + H)+And 292(M-COOC (CH)3)3+H)+。
Intermediate 11: rac-S- (3-aminophenyl) -N-ethoxycarbonyl S-methylsulfoxylimine (A11)
The title compound a11 was prepared from 3-nitroanisole according to the procedure described in WO 2008/006560.
Intermediate 12: [4- ((3- ((4-chloro-1, 3, 5-triazin-2-yl) amino) phenyl) methanesulfonamido) butyl]Carbamic acid tert-butyl ester (A12)
Step 1: to an ice-cold solution of (3-nitrobenzyl) sulfonyl chloride (7.00g,29.7mmol) in dry DCM (60mL) was added pyridine (4mL,3.91g,49.4mmol) and N-tert-butoxycarbonyl-1, 4-diaminobutane (5.60g,29.7mmol) in order. Mixing at RTThe material was stirred for 5.5 hours, diluted with CDM, washed with water, MgSO4Dried and concentrated under reduced pressure. Treated with toluene and the solvent removed again. After stirring with diethyl ether (200mL), the sulfonamide solidified. It was collected by filtration and dried under vacuum; yield: 5.90g (51%).
Step 2: to a solution of the nitro compound of step 1 (5.90g, 15.2mmol) in methanol (250mL) was added Raney nickel (2 g). The mixture was hydrogenated in a Parr reactor at 50 ℃ and 5bar of hydrogen for 36 hours. The catalyst was removed by filtration through celite and the filtrate was concentrated to dryness under reduced pressure to give crude aniline, which was used in the next step without further purification; yield: 5.0g (92%).
Step 3: a solution of the aniline of the previous step (2.75g,7.7mmol) in a THF/iPrOH1:1(5mL) mixture was cooled to-20 ℃. After the addition of DIPEA (2.6mL), a solution of 2, 4-dichloro-1, 3, 5-triazine (1.15g,7.7mmol) in THF/iPrOH (10mL) was added dropwise over 30 minutes. The mixture was stirred at-20 ℃ for a further 1h, the solvent was removed in vacuo and the crude chlorotriazine A12 was used in the next step without further purification.
Intermediate 13: 4-chloro-N- (3- (methylsulfonyl) phenyl) -1,3, 5-triazin-2-amine (A13)
A solution of 3- (methylsulfonyl) aniline hydrochloride (277mg,1.3mmol) in THF/iPrOH1:1(3mL) was cooled to-30 deg.C and treated with DIPEA (517mg, 665. mu.L, 4 mmol). The mixture was added dropwise to a cooled solution of 2, 4-dichloro-1, 3, 5-triazine (200mg,1.33mmol) in THF/iPrOH1:1(3mL) at-30 ℃. It was stirred at-30 ℃ for a further 1h, concentrated to dryness in vacuo and the crude intermediate a13 was used in the next step without further purification.
Intermediate 14: 4- [ (4-chloro-1, 3, 5-triazin-2-yl) amino group]Benzomethanesulfonamide (A14)
A14 was prepared from (4-aminophenyl) methanesulfonyl chloride (250mg,1.3mmol) and 2, 4-dichloro-1, 3, 5-triazine (200mg,1.3mmol) in the presence of DIPEA (345mg, 440. mu.L, 2.7mmol) according to the procedure reported for A13.
EXAMPLES Compounds
Example 1: 3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B1)
With N2Stream A1(1.0eq), 2-methoxyphenylboronic acid (1.5eq) and K3PO4(2.0eq) the mixture in dioxane/water (50/1,0.1M) was degassed for 15 min. Adding Pd (dppf) Cl2(0.1eq), the reaction mixture was heated to 140 ℃ in a microwave oven and maintained for 1 h. The mixture was concentrated under reduced pressure and H was used2The residue was purified by reverse phase RP-HPLC (column: C18) using O (0.1% TFA) and MeOH (0.1% TFA) as eluents. The desired fraction was lyophilized to obtain the title compound (B1) (2%) as a white powder.1HNMR(400MHz,d6-DMSO,300K)3.87(s,3H),4.25(s,2H),6.87(s,2H),7.10(m,2H),7.21(d,J=8.4Hz,1H),7.37(t,J=8.0Hz,1H),7.55(m,2H),7.71(s,1H),7.84(m,1H),8.82(s,1H),10.52(s,1H).MS(ES)C17H17N5O3Theoretical value of S: 371, measured value: 372(M + H)+。
Examples 2-23 were prepared by essentially the same method as described for B1.
Example 2: 3- [ (4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B2)
B2 was prepared according to the general procedure reported for B1 using a1 and 4-fluoro-2-methoxyphenylboronic acid. MS (ES) C17H16FN5O3Theoretical value of S: 389, measurement: 390(M + H)+。
Example 3: 3- [ (4- (5-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B3)
B3 was prepared according to the general procedure reported for B1 using a1 and 5-fluoro-2-methoxyphenylboronic acid. MS (ES) C17H16FN5O3Theoretical value of S: 389, measurement: 390(M + H)+。
Example 4: 3- [ (4- (6-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B4)
B4 was prepared according to the general procedure reported for B1 using a1 and 6-fluoro-2-methoxyphenylboronic acid. MS (ES) C17H16FN5O3Theoretical value of S: 389, measurement: 390(M + H)+。
Example 5: 3- [ (4- (3, 5-difluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B5)
B5 was prepared according to the general procedure reported for B1 using a1 and 3, 5-difluoro-2-methoxyphenylboronic acid. MS (ES) C17H15F2N5O3Theoretical value of S: 407, measurement value: 408(M + H)+。
Example 6: 3- [ (4- (4-chloro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B6)
B6 was prepared according to the general procedure reported for B1 using a1 and 4-chloro-2-methoxyphenylboronic acid. MS (ES) C17H15ClN5O3Theoretical value of S: 405, measurement value: 406(M + H)+。
Example 7: 3- [ (4- (5-chloro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B7)
Using A1 and 5-chloro-2-methoxyphenylboronic acid according to p-B1General procedure reported preparation B7. MS (ES) C17H15ClN5O3Theoretical value of S: 405, measurement value: 406(M + H)+。
Example 8: 3- [ (4- (2-methoxy-4-trifluoromethylphenyl) -1,3, 5-triazin-2-yl) amino group]Benzomethanesulfonamide (B8)
B8 was prepared according to the general procedure reported for B1 using a1 and 2-methoxy-4-trifluoromethylphenylboronic acid. MS (ES) C18H16F3N5O3Theoretical value of S: 439, measured value: 440(M + H)+。
Example 9: 3- [ (4- (2-methoxy-5-trifluoromethylphenyl) -1,3, 5-triazin-2-yl) amino group]Benzomethanesulfonamide (B9)
B9 was prepared according to the general procedure reported for B1 using a1 and 2-methoxy-5-trifluoromethylphenylboronic acid. MS (ES) C18H16F3N5O3Theoretical value of S: 439, measured value: 440(M + H)+。
Example 10: 3- [ (4- (5-hydroxymethyl-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B10)
B10 was prepared according to the general procedure reported for B1 using a1 and 5-hydroxymethyl-2-methoxyphenylboronic acid. MS (ES) C18H19N5O4Theoretical value of S: 401, measurement value: 402(M + H)+。
Example 11: 3- [ (4- (5-formyl-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B11)
Using A1 and 5-formyl 2-methoxyphenylboronic acid, according to the general procedure reported for B1Preparation B11. MS (ES) C18H17N5O4Theoretical value of S: 399, measured value: 400(M + H)+。
Example 12: 3- [ (4- (2-ethoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B12)
B12 was prepared according to the general procedure reported for B1, using a1 and 2-ethoxyphenylboronic acid. MS (ES) C18H19N5O3Theoretical value of S: 385, measurement value: 386(M + H)+。
Example 13: 3- [ (4- (2-benzyloxyphenyl) -1,3, 5-triazin-2-yl) amino group]Benzomethanesulfonamide (B13)
B13 was prepared according to the general procedure reported for B1 using a1 and 2-benzyloxyphenylboronic acid. MS (ES) C23H21N5O3Theoretical value of S: 447, measured value: 448(M + H)+。
Example 14:1- (3- { [4- (2-phenoxyphenyl) -1,3, 5-triazin-2-yl]Amino } phenyl) methanesulfonamide (B14)
B14 was prepared according to the general procedure reported for B1, using a1 and 2-phenoxyphenylboronic acid.1HNMR(400MHz,d6-DMSO,300K)4.17(s,2H),6.83(m,2H),6.91(m,2H),7.03(m,4H),7.29(m,3H),7.52(m,2H),7.93(br,2H),8.72(s,1H),10.31(s,1H)。
Example 15: 3- [ (4- (1, 3-benzodioxol-4-yl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B15)
B15 was prepared according to the general procedure reported for B1, using a1 and (1, 3-benzodioxol-4-yl) boronic acid. MS (ES) C17H15N5O4Theoretical value of S: 385, measurement value: 386(M + H)+。
Example 16: 3- [ (4- (2- ((4-pyridinyl) methoxy) phenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B16)
B16 was prepared according to the general procedure reported for B1, using a1 and a 9. MS (ES) C22H20N6O3Theoretical value of S: 448, measured value: 449(M + H)+。
Example 17: 3- [ (4- (2- (4- (tert-butoxycarbonylamino) butoxy) phenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B17)
B17 was prepared according to the general procedure reported for B1, using a1 and a 10. MS (ES) C25H32N6O5Theoretical value of S: 528, measurement: 529(M + H)+。
Example 18: 3- [ (4- (4-methoxypyridin-3-yl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B18)
B18 was prepared according to the general procedure reported for B1 using a1 and 4-methoxy-3-pyridylboronic acid. MS (ES) C16H16N6O3Theoretical value of S: 372, measuring: 373(M + H)+。
Example 19: 3- [ (4- (3-methoxypyridin-4-yl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B19)
B19 was prepared according to the general procedure reported for B1 using a1 and 3-methoxy-4-pyridylboronic acid. MS (ES) C16H16N6O3Theoretical value of S: 372, measuring: 373(M + H)+。
Example 20: 3- [ (4- (2- ((morpholin-4-yl) methyl) phenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B20)
B20 was prepared according to the general procedure reported for B1 using a1 and (2- ((morpholin-4-yl) methyl) phenyl) boronic acid. MS (ES) C21H24N6O3Theoretical value of S: 440, measured value: 441(M + H)+。
Example 21: 3- [ (4- (2- ((piperidin-1-yl) methyl) phenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B21)
B21 was prepared according to the general procedure reported for B1 using a1 and (2- ((piperidin-1-yl) methyl) phenyl) boronic acid. MS (ES) C22H26N6O2Theoretical value of S: 438, measurement value: 439(M + H)+。
Example 22: 3- [ (4- (2- (cyclopropylaminomethyl) phenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B22)
B22 was prepared according to the general procedure reported for B1 using a1 and (2- (cyclopropylaminomethyl) phenyl) boronic acid. MS (ES) C20H22N6O2Theoretical value of S: 410, measurement value: 411(M + H)+。
Example 23: 3- [ (4- (6-Aminopyridin-3-yl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (B23)
B23 was prepared according to the general procedure reported for B1 using a1 and 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridin-2-amine. MS (ES) C20H22N6O2Theoretical value of S: 357, measurement value: 358(M + H)+。
Example 24: 3- [ (4- (2- (methoxymethyl) phenyl) -1,3, 5-triazin-2-yl) amino group]Benzomethanesulfonamide (B24)
Using a1(400mg,1.33mmol) and (2- (methoxymethyl) phenyl) boronic acid (221mg,1.33mmol), B24 was prepared according to the general procedure reported for B1; yield: 6mg, 1.2%. MS (ES) C18H19N5O3Theoretical value of S: 385, measurement value: 386(M + H)+。
Example 25: 3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzenesulfonamide (C1)
With N2Stream A2(1.0eq), 2-methoxyphenylboronic acid (1.5eq) and K3PO4(2.0eq) the mixture in dioxane-water (50:1,0.1M) was degassed for 15 min. Adding Pd (dppf) Cl2(0.1eq), the reaction mixture was heated to 140 ℃ in a microwave oven and maintained for 1 h. The mixture was concentrated under reduced pressure and H was used2The residue was purified by reverse phase RP-HPLC (column: C18) using O (0.1% TFA) and MeOH (0.1% TFA) as eluents. The desired fractions were lyophilized to give the title compound C1(45%) as a white powder.1HNMR(400MHz,d6-DMSO,300K)3.86(s,3H),7.08(t,J=7.4Hz,1H),7.21(d,J=8.2Hz,1H),7.37(s,2H),7.55(m,4H),7.80(m,1H),8.28(s,1H),8.88(s,1H),10.70(s,1H).MS(ES)C16H15N5O3Theoretical value of S: 357, measurement value: 358(M + H)+。
Example 26: 2- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]Ethanesulfonamide (D1)
With N2Stream A5(1.0eq, 65% purity), 2-methoxyphenylboronic acid (1.5eq) and K3PO4(2.0eq) the mixture in dioxane-water (50:1,0.1M) was degassed for 15 min. Adding Pd (dppf) Cl2(0.1eq), the reaction mixture was heated to 140 ℃ in a microwave oven and maintained for 1 h.The mixture was concentrated under reduced pressure and H was used2The residue was purified by reverse phase RP-HPLC (column: C18) using O (0.1% TFA) and MeOH (0.1% TFA) as eluents. The desired fractions were lyophilized to give the title compound D1(6%) as a white powder.1HNMR(400MHz,d6-DMSO,300K)3.02(m,2H),3.26(m,2H),3.90(s,3H),6.90(br.s,2H),7.04(d,J=7.5Hz,1H),7.12(t,J=7.5Hz,1H),7.22(d,J=8.2Hz,1H),7.32(t,J=7.9Hz,1H),7.58(t,J=7.9Hz,1H),7.68(m,2H),7.85(br.s,1H),8.86(s,1H),10.70(s,1H).MS(ES)C18H19N5O3Theoretical value of S: 385, measurement value: 386(M + H)+。
Example 27: 2- [3- ((4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]Ethanesulfonamide (D2)
D2 was prepared according to the general procedure reported for D1 using a5 and 4-fluoro-2-methoxyphenylboronic acid. MS (ES) C18H18FN5O3Theoretical value of S: 403, measurement value: 404(M + H)+。
Example 28: 3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzamide (E1)
E1 was prepared according to the general procedure reported for B1 using a6 and 2-methoxyphenylboronic acid. Using H2After reverse phase RP-HPLC (column: C18) with O and MeOH as eluents, the desired fractions were lyophilized to give the title compound E1(35%) as a white powder.1HNMR(400MHz,d6-DMSO,300K)3.85(s,3H)6.91(t,J=7.3Hz,1H),6.95(d,J=8.3Hz,1H),7.07(t,J=7.3Hz,1H),7.19(d,J=8.3Hz,1H),7.36(m,1H),7.42(t,J=8.3Hz,1H),7.55(m,2H),7.91(br.s,1H),8.19(s,1H),8.84(s,1H),10.61(s,1H).MS(ES)C17H15N5O2Theoretical value: 321, measurement value: 322(M + H)+。
Practice ofExample 29: 6- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]-2, 3-dihydro-1H-indole-1-sulfonamide (F1)
F1 was prepared according to the general procedure reported for B1, using A8 and 2-methoxyphenylboronic acid. MS (ES) C18H18N6O3Theoretical value of S: 398, measurement: 399(M + H)+。
Example 30: rac-S- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]-N-ethoxycarbonyl-S-methylsulfoxeimide (G1)
The title compound G1 was prepared from a7 and 2-methoxyphenylboronic acid according to the general procedure reported for B1. MS (ES) C20H21N5O3Theoretical value of S: 427, measurement value: 428(M + H)+。
Example 31: 4- (2-methoxyphenyl) -N- (3-nitrophenyl) -1,3, 5-triazin-2-amine (H1)
H1 was prepared according to the general procedure reported for B1 using a3 and 2-methoxyphenylboronic acid.1HNMR(400MHz,d6-DMSO,300K)3.84(s,3H),7.08(d,J=7.5Hz,1H),7.21(d,J=8.3Hz,1H),7.53(t,J=8.7Hz,1H),7.63(t,J=8.3Hz,1H),7.79(br.s,1H),7.90(d,J=7.5Hz,1H),8.24(br.s,1H),8.88(s,1H),8.91(s,1H),10.75(s,1H).MS(ES)C16H13N5O3Theoretical value: 323, measurement value: 324(M + H)+。
Example 32: 3- [ (4- (2- (4-aminobutoxy) phenyl) -1,3, 5-triazin-2-yl) amino group]Benzomethanesulfonamide (I1)
Triazine B17(25mg,0.047mmol) was dissolved in a mixture of TFA (2ml) and DCM (2 ml). Deprotection was complete after 1 hour of stirring at RT as determined by LC/MS. Removing the solvent under reduced pressure and using H2O (0.1% TFA) MeOH (0.1% TFA) asThe residue was purified by reverse phase RP-HPLC (column: C18) for eluent. The desired fraction was lyophilized to give the title compound I1 as a yellow solid. MS (ES) C20H24N6O3Theoretical value of S: 428, measurement: 429(M + H)+。
Example 33: n- (3-aminophenyl) -4- (2-methoxyphenyl) -1,3, 5-triazin-2-amine (J1) in H2A mixture of H1(1.0eq) and Pd/C (10% w/w) in MeOH (0.1M) was stirred at RT under an atmosphere (1atm) for 16H. Passing the mixture throughFiltered and the solvent evaporated under reduced pressure. The crude product was purified by flash chromatography on silica gel (cehex/EtOAc =100:0-0:100) to give the desired product J1(18%) as a yellow solid. MS (ES) C16H15N5Theoretical value of O: 293, measurement value: 294(M + H)+。
Example 34: n- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]Methanesulfonamide (K1)
To a mixture of J1(1.0eq) and anhydrous pyridine (3.0eq) in DCM (0.1M) was slowly added methanesulfonyl chloride (2.0eq) at 0 ℃. The mixture was stirred for 16h, quenched by addition of MeOH and concentrated under reduced pressure. Using H2O (0.1% TFA) and MeOH (0.1% TFA) were used as eluents to purify the residue by reverse phase RP-HPLC (column: C18). The desired fractions were lyophilized to give the title compound K1(44%) as a white powder.1HNMR(400MHz,d6-DMSO,300K)2.96(s,3H),3.83(s,3H),6.90(d,J=8.3Hz,1H),7.06(t,J=7.3Hz,1H),7.17(d,J=8.1Hz,1H),7.28(t,J=8.1Hz,1H),7.51(t,J=8.3Hz,1H),7.63(m,2H),7.76(br.s,1H),8.80(s,1H),9.75(s,1H),10.50(s,1H).MS(ES)C17H17N5O3Theoretical value of S: 371: measurement values: 372(M + H)+.
Example 35: n- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]Propanesulfonamide (L1)
L1 was prepared according to the general procedure reported for K1 using J1 and propane-1-sulfonyl chloride. MS (ES) C19H21N5O3Theoretical value of S: 399, measured value: 400(M + H)+。
Example 36: n- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]Acetamide (M1)
M1 was prepared according to the general procedure reported for K1, using J1 and acetyl chloride.1HNMR(400MHz,d6-DMSO,300K)2.02(s,3H),3.83(s,3H),7.06(t,J=7.8Hz,1H),7.18(d,J=8.9Hz,1H),7.24(t,J=8.1Hz,1H),7.29(m,1H),7.52(t,J=7.8Hz,1H),7.91(s,1H),8.80(s,1H),9.92(s,1H),10.40(s,1H).MS(ES)C18H17N5O2Theoretical value: 335, measured value: 336(M + H)+。
Example 37: n- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]-N' -phenylurea (N1)
A mixture of J1(1.0eq), phenyl isocyanate (2.0eq) and pyridine (3.0eq) in dry DCM (0.01M) was stirred at RT for 12 h. The mixture was concentrated under reduced pressure and dissolved with a minimum amount of DMSO. Water was added and the precipitate was filtered off. The white solid was washed with water and dried in vacuo to give the desired product N1 (62%).1HNMR(400MHz,d6-DMSO,300K)3.83(s,3H),6.97(t,J=7.0Hz,1H),7.06(t,J=7.4Hz,1H),7.18(d,J=7.8Hz,1H),7.28(m,4H),7.46(m,3H),7.51(m,2H),7.80(s,1H),8.60(m,2H),8.80(s,1H),10.26(s,1H).MS(ES)C23H20N6O2Theoretical value: 412, measurement: 413(M + H)+。
Example 38: 3- [ (4- (2-methoxy-5- (methylaminomethyl) phenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (O1)
Sodium borohydride (1.5eq) was added to a mixture of B11 and methylamine solution (MeOH,2M,2.0eq.) in MeOH (0.1M) at 0 ℃. After 2h, the reaction mixture was stirred at RT for a further 12 h. The solution was diluted with EtOAc and sequentially with saturated NaHCO3The organic layer was washed with aqueous solution and brine. With Na2SO4After drying, the solvent was removed in vacuo. Using H2O (0.1% TFA) and MeOH (0.1% TFA) were used as eluents to purify the residue by reverse phase RP-HPLC (column: C18). The desired fractions were lyophilized to give the title compound O1(3%) as a white powder. MS (ES) C19H22N6O3Theoretical value of S: 414, measured value: 415(M + H)+。
Example 39: 4- (2-methoxyphenyl) -N-phenyl-1, 3, 5-triazin-2-amine (P1) A mixture of 2-methoxybenzamide (1.0eq) and N, N-dimethylformamide dimethyl acetal (1.4eq) was heated at 80 ℃ for 1 h. Excess reagent was removed under reduced pressure. The crude product was dissolved in 1, 4-dioxane (0.2M) and then phenylguanidine carbonate (0.43eq) and potassium tert-butoxide (0.41eq) were added. At reflux and N2The reaction mixture was stirred under atmosphere for 12 h. After removal of the solvent, H is used2O (0.1% TFA) and MeOH (0.1% TFA) were used as eluents to purify the residue by reverse phase RP-HPLC (column: C18). The desired fractions were lyophilized to give the title compound P1(20%) as a white powder.1HNMR(400MHz,d6-DMSO,300K)3.87(s,3H),7.08(t,J=7.2Hz,2H),7.20(d,J=8.3Hz,1H),7.35(t,J=7.8Hz,2H),7.54(t,J=7.8Hz,1H),7.81(m,3H),8.81(s,1H),10.50(s,1H).MS(ES)C16H14N4Theoretical value of O: 278, measurement: 279(M + H)+。
Example 40: [4- ((3- ((4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl) methanesulfonylamino) butyl]Carbamic acid tert-butyl ester(Q1)
To a solution of crude intermediate a12(3.62g,7.7mmol) in dioxane/water 50:1(75mL) was added 4-fluoro-2-methoxyphenylboronic acid (1.96g,11.5mmol) and K3PO4(3.26g,15.4 mmol). A nitrogen stream was passed through the mixture for 15 minutes, followed by the addition of Pd (dppf) Cl2(0.63g,0.77 mmol). After an additional 15 minutes of nitrogen, the mixture was stirred at 140 ℃ overnight, cooled to RT and poured into ice-cold water. Extracted with EtOAc and MgSO4The organic layer was dried. Removing solvent under reduced pressure, and performing column chromatography (silica gel, CHCl)3After a MeOH gradient 100:0-90:10) the title compound Q1(1.1g,25%) was obtained as an amorphous solid.1HNMR(300MHz,CDCl3,300K)1.10-1.23(m,2H),1.35(s,9H),1.40-1.48(m,2H),1.53(s,1H),2.90-3.11(m,4H),3.85(s,3H),4.20(s,2H),4.48(bs,1H),6.61-6.72(m,2H),6.92(bs,1H),7.10-7.21(m,1H),7.58(s,1H),7.70-7.97(m,3H),8.56(bs,1H).MS(ES)C26H33FN6O5Theoretical value of S: 560, measurement: 561(M + H)+,461(M+H-COOC(CH3)3)+。
EXAMPLE 41: n- (4-aminobutyl) -1- [3- ((4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl]Methanesulfonamide (R1)
A solution of Q1(1.10g,1.96mmol) in DCM (20mL) was cooled with ice while TFA (2.24g,19.6mmol) was added dropwise over 15 min. After stirring at RT for 2 hours, the solvent was removed under reduced pressure. The residue was dissolved in EtOAc and taken up with saturated NaHCO3The organic solution was washed with aqueous solution and MgSO4Dried and concentrated in vacuo. By column chromatography (silica gel, CHCl containing 1% triethylamine)3After purification in MeOH gradient 9:1-5:1) the title compound R1(66mg,7%) was obtained as a white powder.1HNMR(300MHz,MeOD,300K)1.28-1.50(m,2H),1.50-1.68(m,2H),2.71-2.82(m,2H),2.82-2.97(m,2H),3.80(s,3H),4.21(s,2H),6.66-6.78(m,1H),6.81-6.92(m,1H),7.02-7.12(m,1H),7.20-7.36(m,1H),7.60-7.90(m,3H),8.58(s,1H).MS(ES)C21H25FN6O3Theoretical value of S: 460, measurement: 461(M + H)+。
Example 42: 4- (2-methoxyphenyl) -N- (3- (methylsulfonyl) phenyl) -1,3, 5-triazin-2-amine (S1)
S1 was prepared from crude A13(370mg,1.3mmol) and 2-methoxyphenylboronic acid (303mg,2mmol) according to the procedure described for Q1. The title compound S1 was purified by thick layer chromatography (silica gel, chloroform/MeOH 9:1) to give 54mg of a crop of product, which was purified by reverse phase RP-HPLC (column: C18; H containing 0.1% TFA)2O/MeOH gradient) was further purified; yield: 1.8mg, white amorphous solid.1HNMR(400MHz,CDCl3,300K)3.08(s,3H),3.94(s,3H),7.04-7.11(m,2H),7.50(dt,J=7.4Hz,J=1.8Hz,1H),7.56(t,J=8.0Hz,1H),7.61(bs,1H),7.68(bd,J=7.8Hz,1H),7.90-8.01(bm,2H),8.41(s,1H),8.89(s,1H).MS(ES)C17H16N4O3Theoretical value of S: 356, measurement: 357(M + H)+。
Example 43: 4- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzomethanesulfonamide (T1)
T1 was prepared from crude A14(390mg,1.3mmol) and 2-methoxyphenylboronic acid (303mg,2mmol) according to the procedure described for Q1. The title compound T1 was purified by thick layer chromatography (silica gel, chloroform/MeOH 9:1) to afford a white amorphous solid; yield: 20mg (4%).1HNMR(300MHz,d6-DMSO,300K)3.87(bs,3H),4.21(s,2H),6.82(s,2H),7.07(t,J=7Hz,1H),7.13-7.25(m,1H),7.34(d,J=9Hz,2H),7.47-7.59(m,1H),7.70-7.97(bm,3H),8.82(s,1H),10.38(s,1H).MS(ES)C17H17N5O3Theoretical value of S: 371, measured value: 372(M + H)+。
The following examples 44-53 were prepared by essentially the same method as described for B1.
Example 44:1- [3- ({4- [ 4-fluoro-2- (trifluoromethyl) phenyl)]-1,3, 5-triazin-2-yl } amino) phenyl]Methanesulfonamide (U1)
U1 was prepared according to the general procedure reported for B1 using a1 and [ 4-fluoro-2- (trifluoromethyl) phenyl ] boronic acid.
1HNMR(400MHz,d6-DMSO,300K)4.23(s,2H),6.85(s,2H),7.10(m,1H),7.34(m,1H),7.70(m,3H),7.81(m,1H),7.91(m,1H),8.85(s,1H),10.52(s,1H)。
Example 45: 1- [3- ({4- [ 4-fluoro-2- (propan-2-yloxy) phenyl ] -1,3, 5-triazin-2-yl } amino) phenyl ] methanesulfonamide (U2)
U2 was prepared according to the general procedure reported for B1 using a1 and [ 4-fluoro-2- (prop-2-yloxy) phenyl ] boronic acid.
1HNMR(400MHz,d6-DMSO,300K)1.22(d,6H),4.19(s,2H),4.65(m,1H),6.83(m,3H),7.04(m,2H),7.29(m,1H),7.70(br,3H),8.76(s,1H),10.29(s,1H)。
Example 46:1- (3- { [4- (2-cyano-4-fluorophenyl) -1,3, 5-triazin-2-yl]Amino } phenyl) methanesulfonamide (U3)
U3 was prepared according to the general procedure reported for B1 using a1 and (2-cyano-4-fluorophenyl) boronic acid.1HNMR(400MHz,d6-DMSO,300K)4.25(s,2H),6.85(s,2H),7.10(m,1H),7.34(m,1H),7.73(br,3H),8.01(m,1H),8.40(br,1H),8.88(s,1H),10.54(s,1H)。
Example 47: n- [ 5-fluoro-2- (4- { [3- (sulfamoylmethyl) phenyl]Amino } -1,3, 5-triazin-2-yl) phenyl]Acetamide (U4)
U4 was prepared according to the general procedure reported for B1 using a1 and [2- (acetylamino) -4-fluorophenyl ] boronic acid.
1HNMR(400MHz,d6-DMSO,300K)2.10(br,3H),4.26(s,2H),6.88(s,2H),7.05(m,1H),7.15(m,1H),7.39(m,1H),7.70(br,2H),8.39(m,1H),8.56(m,1H),8.88(s,1H),10.52(s,1H),12.40(br,1H)。
Example 48:1- [3- ({4- [2- (cyclopropylmethoxy) -4-fluorophenyl)]-1,3, 5-triazin-2-yl } amino) phenyl]Methanesulfonamide (U5)
U5 was prepared according to the general procedure reported for B1 using A1 and 2- [2- (cyclopropylmethoxy) -4-fluorophenyl ] -4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan.
1HNMR(400MHz,d6-DMSO,300K)0.29(m,2H),0.46(m,2H),1.15(m,1H),3.95(m,2H),4.22(s,2H),6.84(m,3H),7.06(m,2H),7.33(m,1H),7.74(br,3H),8.79(s,1H),10.31(s,1H)。
Example 49:1- (3- { [4- (3, 4-difluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl]Amino } phenyl) methanesulfonamide (U6)
U6 was prepared according to the general procedure reported for B1 using a1 and (3, 4-difluoro-2-methoxyphenyl) boronic acid.
1HNMR(400MHz,d6-DMSO,300K)3.91(br,3H),4.24(s,2H),6.85(s,2H),7.07(m,1H),7.35(m,2H),7.73(m,3H),8.84(s,1H),10.39(s,1H)。
Example 50:1- (3- { [4- (4, 5-difluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl]Amino } phenyl) methanesulfonamide (U7)
U7 was prepared according to the general procedure reported for B1 using a1 and (4, 5-difluoro-2-methoxyphenyl) boronic acid.
1HNMR(400MHz,d6-DMSO,300K)3.87(s,3H),4.23(s,2H),6.84(s,2H),7.08(m,1H),7.36(m,2H),7.80(br,3H),8.80(s,1H),10.39(s,1H)。
Example 51: 4- (4-fluoro-2-methoxyphenyl) -N- [6- (methylsulfonyl) pyridin-3-yl]-1,3, 5-triazin-2-amine (U8)
DIPEA (0.23ml;1.33mmol) was added to a solution of 2, 4-dichloro-1, 3, 5-triazine in THF/iPrOH (1.30ml;1/1) at-40 ℃. A suspension of 5-amino-2-methylsulfonylpyridine (115mg;0.67mmol; prepared according to H.S. Forrest and J.Walker, J.chem.Soc.,1948, 1939-1945) in THF/iPrOH (0.7ml;1/1) was added. The reaction mixture was stirred at-40 ℃ for 3.5 hours and then at 0 ℃ for 3.5 hours. The solvent was carefully removed using a rotary evaporator (rotovab) while maintaining the temperature of the water bath below 30 ℃ to give crude 4-chloro-N- [6- (methylsulfonyl) pyridin-3-yl ] -1,3, 5-triazin-2-amine (420mg) which was used without further purification.
With N2Stream 86mg of crude product, (4-fluoro-2-methoxyphenyl) boronic acid (76mg;0.45mmol) and K3PO4A mixture (127mg;0.60mmol) in dioxane/water (3ml;20/1) was degassed for 15 min. Adding Pd (dppf) Cl2(123mg;0.15mmol), the reaction mixture is heated to 140 ℃ in a microwave oven and maintained for 1 h. The mixture was concentrated under reduced pressure and H was used2The residue was purified by reverse phase RP-HPLC (column: C18) using O (0.1% HCOOH) and acetonitrile as eluents. The desired fractions were lyophilized to give the title compound as a white solid.
1HNMR(400MHz,d6-DMSO,300K)3.21(s,3H),3.91(s,3H),6.90(m,1H),7.11(m,1H),8.03(m,2H),8.57(m,1H),8.91(s,1H),9.17(br,1H),10.94(s,1H)。
Example 52:1- (3- { [4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl]Amino } phenyl) methanesulfonamide hydrochloride (B2')
1- (3- { [4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl ] amino } phenyl) methanesulfonamide (914mg;2.35mmol) was suspended in 1N aqueous hydrochloric acid (2.35ml) and stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure to give the desired product (980mg;2.30 mmol).
1HNMR(400MHz,d6-DMSO,300K)3.90(s,3H),4.23(s,2H),6.91(m,3H),7.13(m,2H),7.37(m,1H),7.81(m,3H),8.82(s,1H),10.83(s,1H)。
Example 53：3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzenesulfonamide trifluoroacetic acid Salt (B1')
With N2The gas flow converts the 3- [ (4-chloro-1, 3, 5-triazin-2-yl) amino group]Phenylmethanesulfonamide (1.0eq), 2-methoxyphenylboronic acid (1.5eq) and K3PO4(2.0eq) the mixture in dioxane/water (50/1,0.1M) was degassed for 15 min. Adding Pd (dppf) Cl2(0.1eq), the reaction mixture was heated to 140 ℃ in a microwave oven and maintained for 1 h. The mixture was concentrated under reduced pressure and H was used2The residue was purified by reverse phase RP-HPLC (column: C18) using O (0.1% TFA) and MeOH (0.1% TFA) as eluents and the desired fractions were lyophilized to give the title compound (B1') (2%) as a white powder.1HNMR(400MHz,DMSO,300K)3.87(s,3H),4.25(s,2H),6.87(s,2H),7.10(m,2H),7.21(d,J=8.4Hz,1H),7.37(t,J=8.0Hz,1H),7.55(m,2H),7.84(br.s,1H),8.82(s,1H),10.52(s,1H).MS(ES)C17H17N5O3Theoretical value of S: 371, measured value: 372(M + H)+。
Example 54: 3- [ (4- (2-benzyloxyphenyl) -1,3, 5-triazin-2-yl) amino group]Benzenesulfonamide trifluoroacetate salt(B13’)
B13' was prepared according to the general procedure reported for B1, using a1 and 2-benzyloxyphenylboronic acid described in example 53.
1HNMR(400MHz,d6-DMSO,300K)4.22(s,2H),5.24(s,2H),6.85(br.s,2H),7.08(m,2H),7.19-7.34(m,5H),7.41-7.61(m,3H),7.69(s,1H),7.80(m,2H),8.86(s,1H),10.53(s,1H).MS(ES)C23H21N5O3Theoretical value of S: 447, measured value: 448(M + H)+。
Example 55: 3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzenesulfonamide trifluoroacetate salt (C1')
With N2Stream A2(1.0eq), 2-methoxyphenylboronic acid (1.5eq) and K3PO4(2.0eq) the mixture in dioxane/water (50:1,0.1M) was degassed for 15 min. Adding Pd (dppf) Cl2(0.1eq), the reaction mixture was heated to 140 ℃ in a microwave oven and maintained for 1 h. The mixture was concentrated under reduced pressure and H was used2The residue was purified by reverse phase RP-HPLC (column: C18) using O (0.1% TFA) and MeOH (0.1% TFA) as eluents and the desired fractions were lyophilized to give the title compound C1' (45%) as a white powder.
1HNMR(400MHz,d6-DMSO,300K)3.86(s,3H),7.08(t,J=7.4Hz,1H),7.21(d,J=8.2Hz,1H),7.37(s,2H),7.55(m,4H),7.80(m,1H),8.28(s,1H),8.88(s,1H),10.70(s,1H).MS(ES)C16H15N5O3Theoretical value of S: 357, measurement value: 358(M + H)+。
Example 56: 3- [ (4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino]Benzenesulfonamide trifluoroacetate salt(B2”)
With N2The gas stream was mixed with A2(1.0eq), 4-fluoro-2-methoxyphenylboronic acid (1.5eq) and K3PO4(2.0eq) the mixture in dioxane/water (50:1,0.1M) was degassed for 15 min. Adding Pd (dppf) Cl2(0.1eq), the reaction mixture was heated to 140 ℃ in a microwave oven and maintained for 1 h. The mixture was concentrated under reduced pressure and H was used2The residue was purified by reverse phase RP-HPLC (column: C18) using O (0.1% TFA) and MeOH (0.1% TFA) as eluents. The desired fractions were lyophilized to give the title compound B2 "(23%) as a white powder.1HNMR(400MHz,d6-DMSO,300K)3.89(s,3H),4.24(s,2H),6.90(m,3H),7.11(m,2H),7.36(t,J=7.9Hz,1H),7.72(m,1H),7.90(m,2H),8.81(s,1H),10.63(s,1H).MS(ES)C17H16FN5O3Theoretical value of S: 389, measurement: 390(M + H)+。
Materials and methods
1. Determination of binding affinity to CDKs
This protocol describes how a lantha screeneu kinase binding assay can be performed to determine the dissociation constant (K) of the compound of formula (I) with the CDK/cyclin complexd). The principle of this assay is based on the binding and displacement of AlexaFluor 647-labeled tracer bound to the active site of the kinase. Binding of the tracer to the kinase is detected using a Eu-labelled antibody. The tracer and antibody bind simultaneously to the kinase, resulting in the formation of a FRET-signal. Binding of the inhibitor to the kinase competes with binding of the tracer, resulting in reduced FRET.
Table 2: reagent, stock solution concentration and final assay concentration
The compounds of general formula (I) summarized in Table 1 were diluted 1:10 from a10 mM MSMSSO stock solution in a total volume of 15. mu.L DMSO. Pre-dilutions of the compound were then diluted in DMSO in a 1:3 series over 8 steps and vortexed slightly. Each compound was then placed in kinase buffer (HEPES: 20mM, pH: 8.0; MgCl)2:10 mM; DTT: 1 mM; brij-35: 0.01%) at 1:33.33, mixed well and vortexed. For each sample, 5 μ L of the diluted compound was mixed with 5 μ L of tracer working solution (e.g., 60nM tracer 236 in kinase buffer for CDK 2/cyclin a) and 5 μ L of CDK/cyclin/anti-GST-AB-working solution (e.g., 15nM CDK 2/cyclin a, 1:250 dilution of anti-GST-AB in kinase buffer) in wells of a small-volume 384-well plate (Corning incorporated, Corning, ny. usa; cat # 3673). Adjusting tracer concentration to that of CDK/cyclinDissociation constant (K) ofd) It was 30nM for CDK 2/cyclin A, CDK 7/cyclin H and CDK 9/cyclin T1, 20nM for CDK 8/cyclin C and 35nM for CDK 9/cyclin K. For negative controls, 5 μ L of an LDMSO working solution (3% DMSO diluted in kinase buffer) was mixed with 5 μ L of an anti-GST-AB-working solution (e.g., 1:250 dilution of anti-GST-AB in kinase buffer for CDK 2/cyclin a) and 5 μ L of a tracer working solution (e.g., 60nM tracer 236 in kinase buffer for CDK 2/cyclin a) in each well. For positive controls, 5 μ L of LDMSO working solution (3% DMSO diluted in kinase buffer) was mixed with 5 μ L of LCDK/cyclin/anti-GST-AB-working solution (e.g., 15nM CDK 2/cyclin a, 1:250 dilution of anti-GST-AB in kinase buffer) and 5 μ L of tracer working solution (e.g., 60nM tracer 236 in kinase buffer for CDK 2/cyclin a) in each well. Positive and negative controls were calculated from at least 8 different sample wells. The 384-well plates were mixed in a Teleshaker plate blender (Beckmann Coulter, Brea, Calif., USA) at 2000rpm for 40 seconds and incubated for 1h at room temperature before reading. The FRET signal was measured using an Envision spectrophotometer (PerkinElmer, Waltham, MA, USA) with a 50. mu.s delay and 300. mu.s integration time under excitation light at 340nm, emission light at 665nm and 615nm (for the kinase tracer and LanthaScreenEu-AB, respectively). K was determined from the sigmoidal dose-response curve using Quattro Workflow software (Quattro GmbH, Munich, Germany)dThe value is obtained. The results are shown in Table 4.
2. Determination of half-maximal inhibitory concentration on CDKs
This protocol describes how to perform the LanceUltraKinasSelect assay to determine the half-maximal Inhibitory Concentration (IC) of the compound of formula (I) and the CDK/cyclin complex50). The principle of this enzymatic assay is based on phosphorylation of the ULight-peptide substrate. The phosphorylation was detected by using a specific EU-labeled anti-phospho (anti-phospho) peptide antibody. Binding of the EU-labeled anti-phosphopeptide antibody to the phosphorylated ULight-labeled peptide results in the formation of a FRET signal. Binding of inhibitors to kinasesPreventing phosphorylation of the ULight-MBP substrate, resulting in reduced FRET.
Table 3: reagent, stock solution concentration and final assay concentration
The compounds of general formula (I) summarized in Table 5 were diluted 1:10 from a10 mM MSMSSO stock solution in a total volume of 15. mu.L DMSO. Pre-dilutions of the compound were then diluted in DMSO in a 1:3 series over 8 steps and vortexed slightly. Each compound solution was then placed in kinase buffer (HEPES: 50mM, pH: 7.5; MgCl)2:10 mM; EGTA: 1 mM; DTT: 2 mM; tween-20: 0.01%) at 1:20, mix well and vortex. For each sample, 2 μ L of the diluted compound was mixed with 6 μ L of LCDK/cyclin/substrate solution and 2 μ L of LATP solution in wells of a small-capacity 384-well plate (Corning incorporated, Corning, NY. USA; cat # 3673). CDK/cyclins were diluted to appropriate concentrations (see Table 3) and ATP concentrations were adjusted to their IC for CDK/cyclins50Concentrations of 3 μ M for CDK 2/cyclin A, 20 μ M for CDK 1/cyclin B1, 25 μ M for CDK 7/cyclin H and CDK 9/cyclin T1, 55 μ M for CDK 6/cyclin D3, 90 μ M for CDK 4/cyclin D1, and 125 μ M for CDK 9/cyclin K. For negative controls, 2 μ L of LDMSO solution (1% final DMSO assay concentration) was mixed with 6 μ L of substrate solution (50nmulight mbp final assay concentration) and 2 μ LATP solution (appropriate final concentrations see table 3) in each well. For positive controls, 2 μ L of LDMSO solution (1% final DMSO assay concentration) was mixed with 6 μ L LCDK/cyclin/substrate (appropriate final concentration see table 2) and 2 μ L of tracer ATP solution (appropriate final concentration see table 3) in each well. Calculated from at least 8 different sample wellsA positive control and a negative control. The 384-well plates were mixed in a Teleshaker plate blender (Beckmann Coulter, Brea, Calif., USA) at 2000rpm for 40 seconds and incubated for 1h at room temperature before reading. mu.L of assay buffer (Lance assay buffer 1X; EDTA: 20 nM; Eu-anti-P-MBP: see Table 3) was added before the reading. The FRET signal was measured using an Envision spectrophotometer (PerkinElmer, Waltham, MA, USA) with a 50. mu.s delay and 300. mu.s integration time under excitation light at 340nm, emission light at 665nm and 615nm (for the kinase tracer and LanthaScreenEu-AB, respectively). IC was determined from sigmoidal dose-response curves using Quattro Workflow software (Quattro GmbH, Munich, Germany)50The value is obtained. The results are shown in Table 5.
3. Cell assay
3.1 RNA-polymerase IISer2 cell phosphorylation assay:
HCT-116 cells (DSMZ, Braunschweig, Germany) were maintained in McCoy cell culture medium + glutamine (PANBIOTEChGmbH, Aidenbach, Germany) supplemented with 10% fetal bovine serum (PAALaborories GmbH, Pasching, Austria) and allowed to stand at 37 ℃ with 5% CO2And (4) growing. For cell phosphorylation assay, 2x105Cells/well/1 ml cells were seeded in 24-well plates (Greiner Bio-One, Frickenhausen, Germany; catalog # 662160). The compounds of general formula (I) summarized in Table 6 were diluted 1:10 from a10 mM MSMSSO stock solution in a total volume of 15. mu.L DMSO. At 37 deg.C/5% CO2After overnight incubation, 1.5 μ L of compound diluted in DMSO was added to each sample well. Wells containing cells and 0.15% DMSO in culture medium were used as positive controls. Wells without cells in the medium but with 0.15% DMSO were used as negative controls. At 37 deg.C/5% CO2Cells were incubated with the compound for 72 h. Cells were washed with phosphate buffered saline prior to lysis. The phosphorylation of RNA polymerase IISer2 and tubulin levels for normalization were then analyzed using Multi-Array technology (mesoscale discovery, Gaithersburg, Maryland, USA, a combination of antibody-coupled electrochemiluminescence detection and patterned arrays). The manufacturer's instructions were followed and all solutions were purchased from mesoscale discovery. Briefly, the buffer was cleaved at CLB1Cells were lysed by incubation in a wash (Zeptosens, Witterswil, Switzerland; 60. mu.L/well) for 30min and the supernatant removed by centrifugation. For the analysis of RNA polymerase IISer 2-phosphorylation, the cleavage products were diluted 1:50 with MesoScale cleavage buffer to which phosphatase and protease inhibitors were added, and 25. mu.L of each sample was pipetted into the MSDMMulti-Array 96-WellplateImagerHighBindPlate (MesoScaleDiscovery; catalog # L15XB-3/L11XB-3) were placed in wells and incubated for 2h at room temperature. To each well was added 150 μ L of MesoScaleTris wash buffer with 3% w/vMesoScale blocking agent A, and the plates were then sealed and incubated for 1h with vigorous shaking. The plates were washed with 1xTris wash buffer (10 xMesoScale wash buffer diluted 1:10 in distilled water), 25. mu.L of antibody solution (CTD 73E10 antibody from Helmholtz ZentrumMunich, Germany diluted 1:100 in MesoScale Tris wash buffer with 1% w/vMesoScale blocker A added) was added, and the plates were washed three times with 1xTris wash buffer. Add 25. mu.L to each wellSULFO-TAGTMSheep-anti-rat-antibody (mesoscale discovery, catalog # R32AH-1, diluted 1:125 in Tris wash buffer with 1% (w/v) blocking agent a), plates were sealed and incubated at room temperature for 1h with vigorous shaking. Finally, the plates were washed three times with Tris wash buffer, 150 μ l of 2x read buffer (ReadBuffer, mesoscale discovery) was added to each well and the plates were immediately analyzed in a SectorImager from mesoscale discovery. For the determination of the tubulin levels, anti-tubulin antibodies (rabbit; BIODESIGN International, Cat # T59840R, diluted 1: 100) and were used according to the protocol for IISer 2-phosphorylation of RNA polymeraseSULFO-TAGTMSamples were analyzed for goat-anti-rat-antibody (mesoscale discovery, catalog # R32AH-1, diluted 1: 125). By usingTubulin levels RNA polymerase IISer2 phosphorylation was normalized and EC was calculated from 2-fold serial dilutions including 6 concentrations in duplicate using XLFit software (IDBS, Guildford, UK)50The value is obtained. The results are shown in Table 6.
3.2 NF-. kappa.B reporter Gene assay
The cells were maintained in RPMI cell culture medium plus glutamine (PANBIOTECeGmbH, Aidenbach, Germany) supplemented with 10% fetal bovine serum (PAALaborories GmbH, Pasching, Austria) and allowed to stand at 37 ℃ with 5% CO2And (4) growing. Use ofCellLineKitV (Lonza, Basel, Switzerland, Cat # VCA-1003) transfected HEK293 cells grown to 50% confluence. Transfection was performed according to the manufacturer's optimized protocol for transfection of HEK293 cells. Briefly, 2X10 was transfected with 5. mu.g of highly purified plasmid DNA5And (4) cells. Cells were transfected with NF-. kappa.B reporter plasmid (pNFkBluc) and pTALluc was used for control or pMAXGFP was used for transfection control. After transfection, cells were taken up in 500. mu.LRMI 1640 cell culture medium, cultured at 37 ℃ for 1h, and 4.5ml of DMEM without phenol red was added to each transfection. Transfected cells were seeded in 96-well plates (Greiner Bio-One, Frickenhausen, Germany, Cat #655098) at 100. mu.L cell suspension/well and cultured for 48 h. To each well 100 μ L DMEM containing 2x concentrated compound diluted with 10 mdmso stock solution or to control wells 100 μ L DMEM with 0.4% DMSO was added.
In this assay compounds of general formula (I) summarized in Table 6 were used, cells were stimulated with 20ng/ml TNF α and at 37%/5% CO2Plates were incubated for 5 h. Cell culture supernatant was removed to leave 100. mu.L of medium per well, and 100. mu.l of BrightGlo luciferase assay reagent (Promega, Madison, Wis., USA, Cat # E2620) was added and shaken in the dark for 5 minutes. Using a Victor photoelectric colorimeter (Per)Kirnelmer, Waltham, MA, USA). EC was calculated with ExcelFit software (IDBS, Guildford, UK) from 2-fold serial dilutions including at least 10 concentrations in duplicate50The value is obtained. The results are summarized in table 6.
3.3TNF α Release assay
Freshly isolated Peripheral Blood Mononuclear Cells (PBMC) were seeded in 96-well cell culture plates at 100. mu.l per well in 200,000 cells in cell culture medium (DMEM cell culture medium + glutamine from PANBIOTECEH GmbH, Aidenbach, Germany) supplemented with 10% fetal bovine serum (PAALABORIES GmbH, Pasching, Austria) and cultured at 37 ℃ in 5% CO2Incubate overnight. To each well was added 100 μ L of cell culture medium containing 2x concentrated test compound diluted with 10mm mdsso stock solution, or to control wells 100 μ L of LDMEM containing 0.4% DMSO. The compounds of general formula (I) summarized in table 6 were used in this assay. At 37 ℃ with 5% CO2Following 1h incubation, cells were stimulated with 1. mu.g/mLLPS (lipopolysaccharide, Sigma, catalog # L4391-1 MG; 1MG/ml stock solution) or used as a negative control without treatment and at 37 ℃/5% CO2Plates were incubated for 6h next, cell culture plates were centrifuged at 2000rpm for 5 minutes and supernatants were transferred to new 96-well polypropylene plates 25 μ L of supernatant was transferred to 96-well plates of the human TNF α tissue culture kit (mesoscale discovery, Gaithersburg, Maryland, USA) and the level of TNF α was analyzed according to the manufacturer's instructions50The value is obtained. The results are summarized in table 6.
3.4 cell viability assay
HeLa-cells or MDAMB 468-cells or MATUADR, H460, DU145, CACO-2 or B16F10 cells were maintained in RPMI1640 medium or McCoy5A cell medium + glutamine (PANBIOTECEH GmbH, Aidenbach, Germany; orderno. P04-22100; P04-05500) supplemented with 10% fetal bovine serum "Gold" (PAALaboratories GmbH, Pasching, Austria; cat # A15-151). For cell viability assay, 400 out of 25 μ L (Hela)Cells, DSMZ Braunschweig cat # ACC57) or 800 (MDAMB468 cells, ATCC cat # HTB-132) per well the cells were seeded in 384 well plates (Greiner Bio-One, Frickenhausen, Germany; cat # 781080). At 37 deg.C/5% CO2After overnight incubation, 25nL or 75nL of compound was added to each sample well by using the BIOMEKFXP laboratory Automation workstation (BeckmanCoulter, USA). Wells containing cells and 0.1% or 0.3% DMSO in culture medium were used as positive controls, and wells containing cells and 10 μ M staurosporine in culture medium were used as negative controls. At 37 deg.C/5% CO2The cells are then incubated with the compound. For cell viability assay, 25. mu.L of CellTiterGlo reagent (Promega, Madison, USA; Cat. G7573) diluted 1:2 in cell culture medium was added to each well. The 384 well plates were placed on an orbital microplate shaker for 2min and incubated at room temperature for a further 10min to stabilize the luminescence signal. Luminescence was determined by Envision plate reader (PerkinElmer, USA). EC was calculated from 3-fold serial dilutions including at least 8 concentrations in duplicate using ExcelFit software (IDBS, Guildford, UK)50The value is obtained. The results are shown in tables 6 and 7.
3.5CDK2/CycE kinase assay
CDK2/CycE inhibitory activity of compounds of the invention was quantified using the CDK 2/cyceetr-FRET assay described in the following paragraphs.
Recombinant fusion proteins of glutathione S-transferase (GST) and human CDK2 and GST and human CycE expressed in insect cells (Sf-9) and purified by glutathione-agarose affinity chromatography were purchased from proqinase gmbh (Freiburg, Germany). Biotinylated peptide biotin-Ttds-YISPLKSPYKISEG (C-terminus in amide form) available from, for example, JERINI Peptidetechnologies, Inc. (Berlin, Germany) was used as a substrate for the kinase reaction.
For this assay, 50nl of a 100-fold concentrated solution of the test compound in DMSO was pipetted into the wells of a black, small-volume 384-well microplate (Greiner Bio-One, Frickenhausen, Germany), CDK2/CycE was added in an aqueous assay buffer [50mM Tris/HCl pH8.0, 10mM MgCl [ ]21.0mM dithiothreitol, 0.1mM sodium orthovanadate, 0.01% (v/v) Nonidet-P40(Sigma)]2. mu.l of the solution of (1). The mixture was incubated at 22 ℃ for 15min to allow the test compound to bind to the enzyme in advance before the kinase reaction started. The kinase reaction was then initiated by adding 3. mu.l of a solution of adenosine triphosphate (ATP, 16.7. mu.M, giving a final concentration of 10. mu.M for a 5. mu.l assay volume) and substrate (1.25. mu.M, giving a final concentration of 0.75. mu.M for a 5. mu.l assay volume) in assay buffer. The resulting mixture was incubated at 22 ℃ for 25 minutes. The concentration of CDK2/CycE was adjusted according to the activity of the enzyme batches and was appropriately selected to perform the assay within the linear range of the assay. Typical concentrations are in the range of 130 ng/ml. Detection reagent (0.2. mu.M streptavidin-XL and from BDPharmingen [ # 558389) was detected by addition of TR-FRET]1nM anti-RB (pSer807/pSer811) -antibody and 1.2 nMLLANCEU-W1024 labeled anti-mouse IgG antibody [ Perkin-Elmer, cat # AD0077]) The reaction was stopped with 5. mu.l of a solution in EDTA-water (100mM EDTA in 100mM HEPES/NaOH, 0.2% (w/v) bovine serum albumin, pH 7.0).
The resulting mixture was incubated at 22 ℃ for 1 hour to phosphorylate the complex between the biotinylated peptide and the detection reagent. The amount of phosphorylated substrate is then assessed by measuring the resonance energy transfer from the Eu-chelate to streptavidin-XL. Thus, the fluorescence luminescence at 620nm and 665nm after excitation at 350nm is determined in a high time resolution fluorescence analysis (HTRF) reader such as Rubystar (BMGLAbtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer). The ratio of the luminescence at 665nm and 622nm was taken as a measure of the amount of phosphorylated substrate. The data were normalized, i.e., enzyme reaction without inhibitor corresponded to 0% inhibition, and the absence of enzyme but the presence of all other assay components corresponded to 100% inhibition. Computing ICs by 4-parameter fitting using self-service software50The value is obtained.
3.6CDK9/CycT1 kinase assay
CDK9/CycT 1-inhibitory activity of the compounds of the invention was quantified using the CDK9/CycT1TR-FRET assay described in the following paragraphs.
Recombinant full-length His-tagged human CDK9 and CycT1 expressed in insect cells and purified by Ni-NTA affinity chromatography were purchased from Invitrogen (catalog No. PV 4131). The biotinylated peptide biotin-Ttds-YISPLKSPYKISEG (C-terminus in amide form) available from, for example, JERINI Peptidetechnologies, Inc. (Berlin, Germany) was used as a substrate for the kinase reaction.
For this assay, 50nl of a 100-fold concentrated solution of the test compound in DMSO was pipetted into a black, small-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), CDK9/CycT1 was added in aqueous assay buffer [50mM Tris/HCl pH8.0, 10mM MgCl [ ]21.0mM dithiothreitol, 0.1mM sodium orthovanadate, 0.01% (v/v) Nonidet-P40(Sigma)]2. mu.l of the solution of (1). The mixture was incubated at 22 ℃ for 15min to allow the test compound to bind to the enzyme in advance before the kinase reaction started. The kinase reaction was then initiated by adding 3. mu.l of a solution of adenosine triphosphate (ATP.16.7. mu.M, giving a final concentration of 10. mu.M for a 5. mu.l assay volume) and substrate (1.25. mu.M, giving a final concentration of 0.75. mu.M for a 5. mu.l assay volume) in assay buffer. The resulting mixture was incubated at 22 ℃ for a reaction time of 25 minutes. The concentration of CDK9/CycT1 was adjusted according to the activity of the enzyme batches and was appropriately selected to perform the assay within the linear range of the assay. Typical concentrations are in the range of 1. mu.g/ml. Detection reagent (0.2. mu.M streptavidin-XL and from BDPharmingen [ # 558389) was detected by addition of TR-FRET]1nM anti-RB (pSer807/pSer811) -antibody and 1.2 nMLLANCEU-W1024 labeled anti-mouse IgG antibody [ Perkin-Elmer, cat # AD0077]) The reaction was stopped with 5. mu.l of a solution in EDTA-water (100mM EDTA in 100mM HEPES/NaOH, 0.2% (w/v) bovine serum albumin, pH 7.0).
The resulting mixture was incubated at 22 ℃ for 1 hour to allow the phosphorylated biotinylated peptide to form a complex with the detection reagent. The amount of phosphorylated substrate is then assessed by measuring the resonance energy transfer from the Eu-chelate to streptavidin-XL. Thus, the fluorescence emission at 620nm and 665nm after excitation at 350nm is determined in an HTRF reader such as Rubystar (BMGLAbtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer). The ratio of the luminescence at 665nm and 622nm was taken as a measure of the amount of phosphorylated substrate. Data were normalized, i.e., enzyme reaction without inhibitor corresponded to 0% inhibition, absenceEnzyme but the presence of all other assay components corresponds to 100% inhibition. Computing ICs by 4-parameter fitting using self-service software50The value is obtained.
4. And (3) biological determination: proliferation assay
Cultured tumor cells (NCI-H460, human non-small cell lung cancer cell, ATCCHTB-177; DU145, hormone-dependent human prostate cancer cell, ATCCHTB-81, HeLa-MaTu, human cervical cancer cell, EPO-GmbH, Berlin; HeLa-MaTu-ADR, multidrug-resistant human cervical cancer cell, EPO-GmbH, Berlin; HeLa human cervical cancer cell, ATCCCL-2; Caco-2 human colorectal cancer, ATCCHTB-37; B16F10 mouse melanoma cell, ATCCRL-6475) were cultured at 5,000 cells/well (DU145, HeLa-MaTu-ADR), 3,000 cells/well (NCI-H460, HeLa-MaTu, HeLa), a density of 1,500 cells/well (Caco-2) or 1,000 cells/well was seeded in 200. mu.L of growth medium supplemented with 10% fetal bovine serum in a 96-well multi-titer plate (multiwell plate). After 24 hours, the cells of one plate (zero plate) were stained with crystal violet (see below) while the medium in the other plate was replaced with fresh medium (200. mu.l) to which the test substance was added at various concentrations (0. mu.M and 0.001-10. mu.M; final concentration of solvent dimethylsulfoxide 0.5%). The cells are cultured for 4 days in the presence of the test substance. Cell proliferation was determined by staining the cells with crystal violet: the cells were fixed for 15 minutes at room temperature by adding 20. mu.l/measurement point of 11% glutaraldehyde solution. After three cycles of washing the fixed cells with water, the plates were dried at room temperature. Cells were stained by adding 100. mu.l/measurement point of 0.1% crystal violet solution (pH 3.0). After three cycles of washing the stained cells with water, the plates were dried at room temperature. The dye was dissolved by adding 100. mu.l/measurement point of 10% acetic acid solution. The extinction was determined photometrically at a wavelength of 595 nm. The change in cell number, in percent, was calculated by normalizing the measurements to the extinction value (=0%) of the zero-point plate and the extinction value (=100%) of the untreated (0 μm) cells. Determination of IC by 4-parameter fitting Using the own software of the company50The value is obtained.
As a result:
1. determination of binding affinity to CDKs
Dissociation constant K for binding of compounds of the invention to CDK9, CDK8, CDK7 and CDK2, respectivelydSummarized in table 4. Comparison of the binding constants of a particular compound of formula (I) with a number of different CDKs shows that the compound always binds more strongly to CDK9 than to other CDKs. Thus, the compounds of formula (I) interact specifically with CDK9, at least selectively with CDK 9.
Table 4: affinity of the compounds of the invention for CDK9, CDK8, CDK7 and CDK2
All values were determined using the assay described in section 2 of the materials and methods section; "n.t." means that the compound was not detected in the assay.
Compound No. CDK 9/cyclin T1 and CDK 9/cyclin K
④:CDK8⑤:CDK7⑥:CDK2
2. Determination of half-maximal inhibitory concentration on CDKs in enzymatic assays
Half maximal Inhibition Constants (IC) for inhibition of CDK9, CDK1, CDK2, CDK4, CDK6 and CDK7, respectively, in table 550) The values show the inhibitory activity of the compounds of the invention.
Table 5: inhibition of CDK9, CDK1, CDK2, CDK4, CDK6 and CDK7 by compounds of the invention
Determination in nM using the LANCE assay described in section 2 of the materials and methods section
All numerical values. "n.t." means that the compound was not detected in the assay.
Compound No. CDK9 No. CDK1 No. CDK2
⑤:CDK4⑥:CDK6⑦:CDK7
Table 5 a: inhibition of CDK9 and CDK2 by compounds of the invention
IC in nM or μ M50(inhibitory concentration at 50% maximal effect) value, "n.t." means that the compound was not detected in the assay.
Compound number
② CDK 9: CDK9/CycT1 kinase assay described in section 3.6 of materials and methods section
③ CDK 2: CDK2/CycE kinase assay as described in section 3.5 of materials and methods section
Reference Ref1-Ref 5:
ref 1: example 85 of WO2008129080a1 page 45
Ref 2: example 49 of WO2008129080a1 page 38
Ref 3: example 33 of WO2003/037346A1, page 32
Ref 4: example 20 of WO2008129080a1 page 32
Ref 5: page 32 example 152 of US20040116388
3. Cell assay
In Table 6 the half maximal Inhibition Constants (IC) for LPS-induced TNF α release in PBMC, NF-. kappa.B reporter activation, cellular CDK9 activity (RNA polymerase IISer2 phosphorylation) and cell viability in HeLa-cells or MDAMB 468-cells, respectively, are given50) The values show the cell viability of the compounds of the invention. The half maximal Inhibition Constant (IC) in Table 7 for cell viability in MaTu/ADR, H460, DU145, CACO-2 or B16F10 cells50) The values show the cell viability of the compounds of the invention.
TABLE 6 LPS-induced TNF α release in PBMCs, NF-. kappa.B reporter activation, cellular CDK9 activity (RNA polymerase IISer2 phosphorylation) and inhibition of cell viability in MDAMB 468-cells by the compounds of the invention all IC's are expressed in nM50(inhibitory concentration at 50% maximal effect); "n.t." means that the compound was not detected in the assay.
Compound number
② TNF alpha Release
③ NF-kB activation
(iv) phosphorylation by RNA polymerase IISer2
(v) cell viability-MDAMB 468 cells
Table 7: inhibition of cell viability of HeLa, MaTu/ADR, H460, DU145, CACO-2 and B16F10 cells by compounds of the invention.
All ICs are expressed in μ M50(inhibitory concentration at 50% maximal effect) value; "n.t." means that the compound was not detected in the assay.
Compound number
② cell Activity-HeLa cells
③ cell Activity-MaTu/ADR cells
Fourthly, the cell activity-H460 cell
Fifth, cell viability-DU 145 cell
Sixthly, cell activity-CACO-2 cell
Seventhly, cell vitality-B16F 10 cells
Claims (8)
1. A compound of the general formula (I)
Wherein
R1To represent
Wherein
L is a bond, -CH2-、-CH2CH2-or-CF2-；
R3is-SO2NH2、-SO2NH(CH3)、-SO2N(CH3)2、-SO2NH(CH2CH2OCH3)、-NHSO2CH3、-NHSO2CH2CH3、-NHSO2CH2CH2CH3、-NHSO2CF3、-SO2CH3、-NHSO2NH2、-SO(NH)CH3；
R4is-H;
R2to represent
Wherein the group-B-Y-R84-R85is-OCH3、-OCH2CH3、-OCH2CH2CH3、-OCH2CH2CH2CH3、-OCH(CH3)2、-OPh、-OCH2Ph、-OCH2(4-pyridyl);
R83is-H, -F or-Cl;
x is 0, 1 or 2;
and acid salt forms and pharmaceutically acceptable salts thereof.
2. The compound of claim 1, wherein
R1To represent
Wherein
substituent-L-R3is-CH2SO2NH2、-CH2CH2SO2NH2、-CF2SO2NH2、-CH2NHSO2NH2、-CH2SO(NH)CH3；
R4is-H;
R2represents 2-methoxyphenyl, 4-fluoro-2-methoxyphenyl or 6-fluoro-2-methoxyphenyl.
3. The compound of claim 1, wherein
R1To represent
Wherein the substituent-L-R3is-CH2SO2NH2，
R4is-H;
R2represents 2-methoxyphenyl, 4-fluoro-2-methoxyphenyl or 2-benzyloxyphenyl,
or a salt thereof.
4. The compound of claim 1, wherein the compound is selected from the group consisting of:
3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (5-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (6-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (3, 5-difluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (4-chloro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (5-chloro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (2-ethoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
3- [ (4- (2-benzyloxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide,
1- (3- { [4- (2-phenoxyphenyl) -1,3, 5-triazin-2-yl ] amino } phenyl) methanesulfonamide,
3- [ (4- (2- ((4-pyridyl) methoxy) phenyl) -1,3, 5-triazin-2-yl) amino ] methanesulfonamide,
3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide,
2- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl ] ethanesulfonamide,
2- [3- ((4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl ] ethanesulfonamide,
N- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl ] methanesulfonamide,
N- [3- ((4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino) phenyl ] propanesulfonamide,
4- (2-methoxyphenyl) -N- (3- (methylsulfonyl) phenyl) -1,3, 5-triazin-2-amine,
4- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] phenylmethanesulfonamide,
1- [3- ({4- [ 4-fluoro-2- (propan-2-yloxy) phenyl ] -1,3, 5-triazin-2-yl } amino) phenyl ] methanesulfonamide,
1- [3- ({4- [2- (cyclopropylmethoxy) -4-fluorophenyl ] -1,3, 5-triazin-2-yl } amino) phenyl ] methanesulfonamide,
1- (3- { [4- (3, 4-difluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl ] amino } phenyl) methanesulfonamide,
1- (3- { [4- (4, 5-difluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl ] amino } phenyl) methanesulfonamide,
3- [ (4- (2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide trifluoroacetate,
1- (3- { [4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl ] amino } phenyl) methanesulfonamide hydrochloride,
3- [ (4- (4-fluoro-2-methoxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide trifluoroacetate salt,
3- [ (4- (2-benzyloxyphenyl) -1,3, 5-triazin-2-yl) amino ] benzenesulfonamide trifluoroacetate salt.
5. Use of a compound according to any one of claims 1 to 4 for the preparation of a pharmaceutical composition for the prophylaxis and/or treatment of infectious diseases including opportunistic diseases, immune diseases, autoimmune diseases, cardiovascular diseases, cell proliferative diseases, inflammation, erectile dysfunction and stroke.
6. The use of claim 5, wherein the cell proliferative disorder is selected from the group consisting of:
adenocarcinoma, choroidal melanoma, acute leukemia, acoustic schwannoma, ampulla, anal, astrocytoma, basal cell carcinoma, pancreatic, desmoid, bladder, bronchial, breast, burkitt's lymphoma, uterine corpus, CUP-syndrome, colorectal, small intestine, ovarian, endometrial, ependymoma, epithelial cancers, Ewing's tumor, gastrointestinal, gallbladder, uterine, cervical, glioblastoma, gynecological, otorhinolaryngological, hematological, hairy cell leukemia, urinary tract, skin testicular, brain metastasis, testicular, pituitary, carcinoid, kaposi's sarcoma, laryngeal, blastoma, bone, head and neck, colon, craniopharyngioma, oral, central nervous system, liver metastasis, liver metastasis, colon, cervical, rectal, and rectal tumors, central nervous system, liver, bladder, and bladder tumors, and cervical tumors Leukemia, meibomian tumors, lung cancer, lymph node cancer, lymphoma, gastric cancer, malignant melanoma, malignant neoplasia, gastrointestinal malignancy, breast cancer, medulloblastoma, melanoma, meningioma, hodgkin's disease, mycosis fungoides, nasal cancer, schwannoma, neuroblastoma, kidney cancer, renal cell carcinoma, oligodendroglioma, esophageal cancer, osteolytic and osteogenic cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, plasmacytoma, prostate cancer, pharyngeal cancer, rectal cancer, retinoblastoma, vaginal cancer, thyroid cancer, schneeeberg's disease, esophageal cancer, spinoaliims, T-cell lymphoma, thymoma, tubular cancer, ocular tumor, urinary tract cancer, urinary tumor, bladder epithelial cancer, vulva cancer, verrucous, soft tissue tumor, soft tissue sarcoma, Wilm tumor, cervical cancer, tongue cancer, invasive ductal cancer, invasive lobular cancer, ductal carcinoma in situ, In situ lobular carcinoma, small cell lung carcinoma, non-small cell lung carcinoma, bronchial adenoma, pleuropulmonoblastoma, mesothelioma, brain stem glioma, hypothalamic glioma, cerebellar astrocytoma, cerebral astrocytoma, neuroectodermal tumors, pinealoma, uterine sarcoma, salivary gland adenocarcinoma, anal gland adenocarcinoma, mastocytoma, pelvic tumor, ureteroma, hereditary papillary renal carcinoma, sporadic papillary renal carcinoma, intraocular melanoma, hepatocellular carcinoma, cholangiocarcinoma, mixed hepatocellular cholangiocarcinoma, squamous cell carcinoma, malignant melanoma, merkel cell skin carcinoma, non-melanoma skin carcinoma, hypopharynx cancer, nasopharyngeal carcinoma, oropharyngeal carcinoma, oral cavity cancer, squamous cell carcinoma, oral melanoma, oromelanoma, AIDS-related lymphoma, cutaneous T-cell lymphoma, central nervous system lymphoma, malignant fibrous histiocytoma, lymphosarcoma, rhabdomyosarcoma, neuroblastoma, melanoma, sarcoma, melanoma, and melanoma, Malignant histiocytosis, fibrosarcoma, angiosarcoma, hemangioendothelioma, leiomyosarcoma, canine breast cancer, and feline breast cancer.
7. A pharmaceutical composition comprising as active ingredient at least one compound according to any one of claims 1 to 4 and at least one pharmaceutically acceptable carrier, excipient and/or diluent.
8. The pharmaceutical composition of claim 7, further comprising one or more other anti-neoplastic agents.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP10075131 | 2010-03-22 | ||
| EP10075131.2 | 2010-03-22 | ||
| US28276610P | 2010-03-29 | 2010-03-29 | |
| US61/282,766 | 2010-03-29 | ||
| PCT/EP2011/001445 WO2011116951A1 (en) | 2010-03-22 | 2011-03-20 | Pharmaceutically active disubstituted triazine derivatives |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HK1181771A1 HK1181771A1 (en) | 2013-11-15 |
| HK1181771B true HK1181771B (en) | 2017-03-17 |
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