GRAHAM KEITH (DE)
BADER BENJAMIN (DE)
HILLIG ROMAN (DE)
SCHRÖDER JENS (DE)
LIENAU PHILIP (DE)
BRIEM HANS (DE)
WO2019201848A1 | 2019-10-24 | |||
WO2019122129A1 | 2019-06-27 |
Claims 1. A compound of formula (Ia) wherein R1 is selected from ‐H, halogen, ‐OH, ‐CN, ‐NO2, C1‐C6‐alkylsulfanyl, ‐NRaRb, wherein Ra and Rb are independently selected from ‐H or C1‐C6‐alkyl, C1‐C6‐alkyl, C1‐C6‐alkoxy, C2‐C6‐alkenyl, C2‐C6‐alkynyl, C3‐C8‐cycloalkyl, C4‐C8‐cycloalkenyl, 4‐ to 7‐membered heterocycloalkyl, 5‐ to 10 membered heterocycloalkenyl, heterospirocycloalkyl optionally substituted by an oxo‐group (=O), fused heterocycloalkyl optionally substituted by an oxo‐group (=O), bridged heterocycloalkyl optionally substituted by an oxo‐group (=O), phenyl, heteroaryl, C1‐C6‐ haloalkyl, ‐C(=O)OH, ‐C(=O)ORc, wherein Rc stands for C1‐C6‐alkyl, C3‐C6‐alkenyl, C3‐C6‐alkynyl, C3‐C8‐cycloalkyl or C4‐C8‐cycloalkenyl, ‐N=S(=O)(Rd)Re, wherein Rd and Re are independently selected from C1‐C6‐alkyl, C2‐C6‐ alkenyl, C2‐C6‐alkynyl, C3‐C8‐cycloalkyl or C4‐C8‐cycloalkenyl, ‐NH‐C(O)‐C1‐C6‐alkyl, ‐NH‐C(O)‐NRaRb, wherein Ra and Rb are selected independently from a hydrogen atom or a C1‐C6‐alkyl, ‐NH‐(CH2)k‐NH‐C(O)‐C1‐C6‐alkyl, wherein k is 1 or 2, ‐NH‐(CH2)l‐Rf, wherein l is 0, 1 or 2 and Rf stands for a 4‐ to 7‐membered heterocycloalkyl, heteroaryl or C1‐C6‐alkylsulfonyl, whereby in all foregoing definitions the C1‐C6‐alkyl‐, C1‐C6‐alkoxy‐, the 4‐ to 7‐ membered heterocycloalkyl and the heteroaryl can be optionally substituted, one or two or three times, identically or differently, with a halogen atom, hydroxy, oxo (=O), a cyano, nitro, C1‐C6‐alkyl, C2‐C6‐alkenyl, C2‐C6‐alkynyl, C3‐C8‐cycloalkyl, 4‐ to 7‐membered heterocycloalkyl, C1‐C6‐alkoxy, C1‐C6‐haloalkyl, C1‐C6‐haloalkoxy, C1‐ C6‐alkylsulfonyl, phenyl, benzyl, heteroaryl, ‐CH2‐heteroaryl, C3‐C8‐cycloalkoxy. phenyloxy, heteroaryloxy, ‐NH‐C(O)‐C1‐C6‐alkyl or –NRaRb, wherein Ra and Rb are independently selected from a hydrogen atom or C1‐C6‐alkyl, ‐O‐(CH2)z‐phenyl, ‐O(CH2)z‐C4‐C7‐heterocycloalkyl, ‐O(CH2)z‐heteroaryl, wherein z is 0, 1 or 2, and the phenyl, heterocycloalkyl and heteroaryl can optionally be substituted with a group selected from hydroxy, heterocycloalkyl or heterocaclyoalkenyl, which both can be substituted with a methyl‐ and/or oxo‐group, wherein L2a stands for C(O), L2b stands for a bond or C1‐C6‐ alkylene, X2 stands for , and Rx2 stands for or in which a further R1 as defined above can be directly attached to a first R1 equaling C1‐C6‐alkyl, C1‐C6‐alkoxy, C2‐C6‐alkenyl, C2‐C6‐alkynyl, C3‐C8‐cycloalkyl, C4‐C8‐cycloalkenyl, 4‐ to 7‐membered heterocycloalkyl, 5‐ to 10 membered heterocycloalkenyl, heterospirocycloalkyl, fused heterocycloalkyl, bridged heterocycloalkyl, phenyl, heteroaryl, C1‐C6‐haloalkyl, y is 1, 2 or 3; and either both T and V stand for nitrogen or T stands for carbon and V for nitrogen or T for nitrogen and V for carbon; A is selected from the group consisting of C6‐10aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; R2 is each independently selected from the group consisting of C1‐4alkyl, C2‐4alkenyl, C2‐ 4alkinyl, C1‐4haloalkyl, hydroxy‐C1‐4alkyl, hydroxy‐C1‐4haloalkyl, C3‐6cycloalkyl, 3‐6 membered heterocyclyl, hydroxy‐C3‐6cycloalkyl, C1‐4haloalkyl substituted with a 3‐6 membered heterocyclyl, 3‐6 membered heterocyclyl substituted with hydroxy, halogen, ‐NH2, ‐SO2‐C1‐4alkyl and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring; R6 is selected from the group consisting of ‐H, halogen, C1‐4alkyl, C3‐7‐cycloalkyl, C4‐ 7heterocycloalkyl optionally comprising 1 or 2 nitrogen, 1 oxygen or 1 sulphur atom, ‐ O‐C1‐4alkyl, ‐NH2, ‐NH(C1‐4alkyl) or ‐NH(C1‐4alkyl)2, x is 1, 2 or 3; or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 2. A compound according to claim 1, wherein R1 is selected from ‐H, ‐Br, ‐OH, ‐NO2, ‐CH3, , ‐O‐CH3, ‐O‐CH2‐CH3, ‐O‐CH(CH3)2, ‐O‐(CH2)3CH3, ‐O‐(CH2)2CH(CH3)2, ‐O‐CH2‐phenyl, ‐O‐(CH2)2‐O‐CH3, ‐O‐(CH2)2‐S(O)2‐CH3, ‐CH2‐OH, ‐C(CH3)2‐OH, ‐C(O)OH, ‐C(O)OCH3, ‐NH2, ‐NH(CH3), ‐N(CH3)2, , , , ‐NH‐(CH2)2‐NH‐C(O)‐CH3, ‐NH‐(CH2)2‐morpholino, ‐NH‐C(O)‐CH3, ‐NH‐C(O)‐NH‐CH3, ‐NH‐C(O)‐N(CH3)2, ‐NH‐S(O)2‐CH3, ‐N=S(O)(CH3)2, y is 1 or 2; A is selected from the group consisting of C6‐10aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; R2 is each independently selected from the group consisting of C1‐4alkyl, C2‐4alkenyl, C2‐ 4alkinyl, C1‐4haloalkyl, hydroxy‐C1‐4alkyl, hydroxy‐C1‐4haloalkyl, C3‐6cycloalkyl, 3‐6 membered heterocyclyl, hydroxy‐C3‐6cycloalkyl, C1‐4haloalkyl substituted with a 3‐6 membered heterocyclyl, 3‐6 membered heterocyclyl substituted with hydroxy, halogen, ‐NH2, ‐SO2‐C1‐4alkyl and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring x is 1, 2 or 3; or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 3. A compound according to claim 1, wherein A is selected from the group consisting of C6‐10‐aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; R2 is each independently selected from the group consisting of C1‐4‐alkyl, C2‐4‐alkenyl, C2‐4‐ alkinyl, C1‐4‐haloalkyl, hydroxy‐C1‐4‐alkyl, hydroxy‐C1‐4‐haloalkyl, C3‐6‐cycloalkyl, 3‐6 membered heterocyclyl, hydroxy‐C3‐6‐cycloalkyl, C1‐4‐haloalkyl substituted with a 3‐6 membered heterocyclyl, 3‐6 membered heterocyclyl substituted with hydroxy, halogen, ‐NH2, ‐SO2‐C1‐4‐alkyl and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring x is 1, 2 or 3 or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 4. A compound according to claim 1, wherein A is selected from the group consisting of C6‐10aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; x is 1 or 2 R2 is each independently selected from the group consisting of C1‐4‐alkyl, C2‐4‐alkinyl, C1‐4‐ haloalkyl, hydroxy‐C1‐4‐haloalkyl, C1‐4‐haloalkyl substituted with a 3‐6 membered heterocyclyl, halogen, and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 5. A compound according to claim 1, wherein and wherein R3 is selected from the group consisting of C1‐4‐alkyl, C1‐4‐haloalkyl, hydroxy‐C1‐4‐alkyl, hydroxy‐C1‐4‐haloalkyl, C1‐4‐haloalkyl substituted with a 3‐6 membered heterocyclyl, C3‐6‐cycloalkyl, hydroxy‐C3‐6‐cycloalkyl, 3‐6 membered heterocyclyl, 3‐6 membered hydroxy‐heterocyclyl, halogen and ‐SO2‐C1‐4‐alkyl; R4 is selected from the group consisting of hydrogen and ‐NH2, R5 is selected from the group consisting of hydrogen, C1‐4‐alkyl and halogen; or R3 and R5 together with the carbon atoms they are attached form a 5‐6 membered nonaromatic carbocycle, a 5‐6 membered non‐aromatic heterocycle or a 5‐6 membered heteroaryl, wherein the 5‐6 membered non‐aromatic carbocycle, 5‐6 membered nonaromatic heterocycle and 5‐6 membered heteroaryl are all optionally substituted by one or more halogen or by an oxo group or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 6. A compound according to claim 1, wherein R3 is selected from the group consisting of C1‐4‐haloalkyl, hydroxy‐C1‐4‐haloalkyl and C1‐4‐ haloalkyl substituted with a 3‐6 membered heterocyclyl; R4 is hydrogen; R5 is selected from the group consisting of hydrogen, C1‐4‐alkyl and fluorine; or R3 and R5 together with the carbon atoms they are attached form a 5‐6 membered nonaromatic carbocycle, a 5‐6 membered non‐aromatic heterocycle or a 5‐6 membered heteroaryl, wherein the 5‐6 membered non‐aromatic carbocycle, 5‐6 membered nonaromatic heterocycle and 5‐6 membered heteroaryl are all optionally substituted by one or more fluorine or by an oxo group or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 7. A compound according to claim 1, wherein is selected from or a stereoisomer, a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 8. The compound according to claim 1, wherein V is nitrogen and T is carbon or a stereoisomer, a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 9. The compound according to claim 1, wherein y = 1 and R1 is selected from or a stereoisomer, a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 10. The compound according to claim 1, wherein V is nitrogen, T is carbon, y = 1, R1 is selected from and is selected from or a stereoisomer, a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 11. The compound according to claim 1, which is selected from the group consisting of: N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐fluoro‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoro‐phenyl]ethyl]‐2‐methyl‐6‐pyrrolidin‐1‐yl‐pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}‐6‐fluoro‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2‐methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2‐methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(1R)‐1‐[3‐(difluoromethyl)‐2‐methyl‐phenyl]ethyl]‐2‐methyl‐6‐pyrrolidin‐1‐yl‐pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐fluoro‐2‐methyl‐N‐[(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐[(3R)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl]‐6‐fluoro‐2‐methyl‐pyrido[3,4‐d]pyrimidin‐4‐ amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl]‐6‐fluoro‐2‐methyl‐pyrido[3,4‐d]pyrimidin‐4‐ amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐fluoro‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2,8‐dimethylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2,8‐dimethyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one N‐{(3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3S)‐1‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide 6‐ethoxy‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 1‐(3‐{(1R)‐1‐[(6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐yl)amino]ethyl}‐2‐fluorophenyl)‐1,1‐ difluoro‐2‐methylpropan‐2‐ol 6‐ethoxy‐N‐{(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluorophenyl]ethyl}‐6‐ethoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine 6‐ethoxy‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phenyl]ethyl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine 6‐methoxy‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐ amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine 6‐methoxy‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluorophenyl]ethyl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine 2,2‐difluoro‐2‐(2‐fluoro‐3‐{(1R)‐1‐[(6‐methoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl)amino]ethyl}phenyl)ethan‐1‐ol 1,1‐difluoro‐1‐(2‐fluoro‐3‐{(1R)‐1‐[(6‐methoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl)amino]ethyl}phenyl)‐2‐methylpropan‐2‐ol N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2‐hydroxyethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2‐hydroxy‐2‐methylpropyl)‐2‐fluorophenyl]ethyl}amino)‐ 2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(2‐methylphenyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(3‐methylphenyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(4‐methylphenyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2‐fluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐fluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(4‐fluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2‐methoxyphenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐methoxyphenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2‐chlorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐chlorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[4‐({(1RS)‐1‐[2‐(difluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1RS)‐1‐[2‐(difluoromethoxy)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethoxy)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethoxy)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐{3‐[(trifluoromethyl)sulfanyl]phenyl}ethyl]amino}pyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1R)‐1‐[3‐(pentafluoro‐lambda6‐ sulfanyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide methyl 3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]benzoate N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐cyanophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(3‐nitrophenyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide tert‐butyl {3‐[(1RS)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]phenyl}carbamate N‐[(3R)‐1‐(4‐{[(1R)‐1‐(4‐fluoro‐3‐methylphenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,3‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3,4‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,4‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(3,5‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(2,6‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(2,5‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromo‐2‐methylphenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromo‐5‐fluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromo‐4‐fluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromo‐2‐fluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromo‐2‐fluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromo‐2‐methoxyphenyl)ethyl]amino}‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐fluoro‐1‐benzofuran‐7‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide. N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3‐fluoro‐1‐benzofuran‐7‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1RS)‐1‐[2‐(1H‐pyrazol‐1‐yl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1RS)‐1‐[3‐(difluoromethyl)‐1‐methyl‐1H‐pyrazol‐4‐yl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1RS)‐1‐[1‐methyl‐3‐(trifluoromethyl)‐1H‐pyrazol‐4‐ yl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(5‐chloro‐1,3‐thiazol‐2‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1RS)‐1‐[3‐(trifluoromethyl)‐1,2,4‐oxadiazol‐5‐ yl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromopyridin‐3‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(6‐aminopyridin‐2‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[1‐(3‐aminophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl)pyrrolidin‐ 3‐yl]acetamide (mixture of stereoisomers) tert‐butyl {3‐[(1S)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]phenyl}carbamate tert‐butyl {3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]phenyl}carbamate N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3‐aminophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐aminophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3,5‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3,5‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1S)‐1‐(2,6‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,6‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,5‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1S)‐1‐(2,5‐difluorophenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide 3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]benzoic acid N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(hydroxymethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐hydroxyphenyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(2,2‐difluoroethoxy)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐{3‐[(E)‐2‐ethoxyethenyl]phenyl}ethyl]amino}‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine 2,2‐difluoro‐2‐{2‐fluoro‐3‐[(1R)‐1‐{[2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐d]pyrimidin‐ 4‐yl]amino}ethyl]phenyl}ethan‐1‐ol 1,1‐difluoro‐1‐{2‐fluoro‐3‐[(1R)‐1‐{[2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐d]pyrimidin‐ 4‐yl]amino}ethyl]phenyl}‐2‐methylpropan‐2‐ol N‐{(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phenyl]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluorophenyl]ethyl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐ yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐N‐{(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl}‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine 2‐{3‐[(1R)‐1‐({6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]‐2‐fluorophenyl}‐2,2‐difluoroethan‐1‐ol 1‐{3‐[(1R)‐1‐({6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]‐2‐fluorophenyl}‐1,1‐difluoro‐2‐methylpropan‐2‐ol 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐2,6‐ diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐ 2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2‐hydroxyethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2‐hydroxy‐2‐methylpropyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6‐ethyl‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N6‐cyclopropyl‐N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐N6‐(propan‐2‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6‐ethyl‐N6,2‐dimethylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6,2‐dimethyl‐N6‐(prop‐2‐en‐1‐ yl)pyrido[3,4‐d]pyrimidine‐4,6‐diamine N6‐cyclopropyl‐N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6,2‐dimethylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N6‐cyclobutyl‐N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6,2‐dimethyl‐N6‐(propan‐2‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6‐(2‐methoxyethyl)‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(piperidin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N6‐cyclopentyl‐N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(piperazin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐ol (3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐ol N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(morpholin‐4‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐N6‐{[(2RS)‐oxetan‐2‐ yl]methyl}pyrido[3,4‐d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐N6‐[(3R)‐oxolan‐3‐yl]pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6‐(2‐methoxyethyl)‐N6,2‐ dimethylpyrido[3,4‐d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐N6,N6‐di(prop‐2‐en‐1‐ yl)pyrido[3,4‐d]pyrimidine‐4,6‐diamine 6‐[2‐azabicyclo[2.2.1]heptan‐2‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(1‐oxa‐6‐azaspiro[3.3]heptan‐ 6‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(2‐oxa‐6‐azaspiro[3.3]heptan‐ 6‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine N6‐cyclohexyl‐N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine 4‐{[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]amino}pyrrolidin‐2‐one (mixture of stereoisomers) 4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]piperazin‐2‐one 6‐(1,4‐diazepan‐1‐yl)‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3R)‐3‐methylmorpholin‐4‐ yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3S)‐3‐methylmorpholin‐4‐ yl]pyrido[3,4‐d]pyrimidin‐4‐amine (3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐3‐ol (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐3‐ol N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐N6‐(oxan‐4‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐N6‐{[(2R)‐oxolan‐2‐ yl]methyl}pyrido[3,4‐d]pyrimidine‐4,6‐diamine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(3S)‐3‐methoxypyrrolidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6‐[2‐(dimethylamino)ethyl]‐N6,2‐ dimethylpyrido[3,4‐d]pyrimidine‐4,6‐diamine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(thiomorpholin‐4‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐[3‐(difluoromethyl)azetidin‐1‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(3,3‐difluoropyrrolidin‐1‐yl)‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(2,6‐dihydropyrrolo[3,4‐c]pyrazol‐5(4H)‐ yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]piperidine‐4‐carbonitrile N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[hexahydrocyclopenta[c]pyrrol‐2(1H)‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[hexahydropyrrolo[3,4‐c]pyrrol‐2(1H)‐ yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3aR,6aS)‐tetrahydro‐1H‐ furo[3,4‐c]pyrrol‐5(3H)‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(3aRS,6aRS)‐hexahydro‐5H‐furo[2,3‐ c]pyrrol‐5‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(2‐oxa‐6‐azaspiro[3.4]octan‐6‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N6‐cyclohexyl‐N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6,2‐dimethylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine 4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐1,4‐diazepan‐2‐one (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidine‐3‐carboxamide (6R)‐4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐6‐methylpiperazin‐2‐one (6S)‐4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐6‐methylpiperazin‐2‐one N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(3,3‐dimethylpiperazin‐1‐yl)‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(4‐methyl‐1,4‐diazepan‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(4‐ethylpiperazin‐1‐yl)‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(3S)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine {1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐4‐yl}methanol N4‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐N6,2‐dimethyl‐N6‐(oxan‐4‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine 4‐{[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]amino}cyclohexan‐1‐ol (mixture of stereoisomers) (1RS,4SR,5RS)‐2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐2‐azabicyclo[2.2.1]heptane‐5‐carbonitrile (mixture of stereoisomers) N2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐N,N,N2‐trimethylglycinamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(6,7‐dihydropyrazolo[1,5‐a]pyrazin‐ 5(4H)‐yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(5,6‐dihydroimidazo[1,5‐a]pyrazin‐7(8H)‐ yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(5,6‐dihydroimidazo[1,2‐a]pyrazin‐7(8H)‐ yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(1‐methyl‐4,6‐ dihydropyrrolo[3,4‐c]pyrazol‐5(1H)‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐(5,6‐dihydro[1,2,4]triazolo[1,5‐a]pyrazin‐ 7(8H)‐yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐4‐methylpiperidine‐4‐carbonitrile {4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}acetonitrile 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐2,6‐diazaspiro[3.4]octan‐5‐one 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3aS,6aS)‐1‐ methylhexahydropyrrolo[3,4‐b]pyrrol‐5(1H)‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3aRS,6aSR)‐5‐ methylhexahydropyrrolo[3,4‐c]pyrrol‐2(1H)‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3aR,6aR)‐1‐ methylhexahydropyrrolo[3,4‐b]pyrrol‐5(1H)‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(8aS)‐hexahydropyrrolo[1,2‐a]pyrazin‐ 2(1H)‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(8aR)‐hexahydropyrrolo[1,2‐a]pyrazin‐ 2(1H)‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(6‐methyl‐2,6‐ diazaspiro[3.4]octan‐2‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine 6‐(4‐cyclopropylpiperazin‐1‐yl)‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(2‐oxa‐6‐azaspiro[3.5]nonan‐6‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(2‐oxa‐7‐azaspiro[3.5]nonan‐7‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine (3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐3‐methylpyrrolidine‐3‐carboxamide 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]piperidine‐4‐carboxamide 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one (3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐3‐carboxamide (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐3‐carboxamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(cis)‐3,4,5‐trimethylpiperazin‐ 1‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3R,5R)‐3,4,5‐ trimethylpiperazin‐1‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(3S,5S)‐3,4,5‐ trimethylpiperazin‐1‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[3‐(dimethylamino)piperidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[4‐(dimethylamino)piperidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐3‐methylpyrrolidine‐3‐carboxylic acid (mixture of stereoisomers) 4‐{[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]amino}‐1‐methylcyclohexan‐1‐ol (mixture of stereoisomers) 2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐ol 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐3‐(2‐hydroxyethyl)pyrrolidin‐3‐ol (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(3‐methyl‐5,6‐ dihydro[1,2,4]triazolo[4,3‐a]pyrazin‐7(8H)‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]hexahydropyrrolo[1,2‐a]pyrazin‐6(2H)‐one (mixture of stereoisomers) (5RS)‐7‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,7‐diazaspiro[4.4]nonan‐3‐one (mixture of stereoisomers) 6‐[[1,3'‐bipyrrolidin]‐1'‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) 7‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]hexahydro‐3H‐[1,3]oxazolo[3,4‐a]pyrazin‐3‐one (mixture of stereoisomers) 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐4‐methyl‐1,4‐diazepane‐2,3‐dione 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐1,4‐diazepan‐1‐yl}ethan‐1‐one N‐{(3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐N‐methylacetamide (mixture of stereoisomers) N‐{1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐4‐yl}acetamide (3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐N‐methylpiperidine‐3‐carboxamide (mixture of stereoisomers) 2‐{1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐4‐yl}propan‐2‐ol (2R)‐4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐6‐oxopiperazine‐2‐carboxylic acid N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[4‐(2‐methoxyethyl)piperazin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine 5‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐4,5,6,7‐tetrahydropyrazolo[1,5‐a]pyrazine‐2‐carbonitrile 6‐[4‐(2,2‐difluoroethyl)piperazin‐1‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[5‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]hexahydropyrrolo[3,4‐c]pyrrol‐2(1H)‐yl]ethan‐1‐one (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[3‐(piperidin‐1‐yl)pyrrolidin‐1‐ yl]pyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[3‐(morpholin‐4‐yl)pyrrolidin‐ 1‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) 6‐[7,7‐difluorohexahydropyrrolo[1,2‐a]pyrazin‐2(1H)‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐ fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) (3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐3‐sulfonamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[4‐(2,2,2‐ trifluoroethyl)piperazin‐1‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine tert‐butyl {(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}carbamate tert‐butyl {3‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐3‐azabicyclo[3.1.0]hexan‐1‐yl}carbamate (mixture of stereoisomers) tert‐butyl {1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐4‐fluoropyrrolidin‐3‐yl}carbamate (mixture of stereoisomers) tert‐butyl 6‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octane‐2‐carboxylate tert‐butyl 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,7‐diazaspiro[3.5]nonane‐7‐carboxylate tert‐butyl 7‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,7‐diazaspiro[3.5]nonane‐2‐carboxylate N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐(6‐methyl‐2,6‐ diazaspiro[3.4]octan‐2‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine tert‐butyl 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octane‐6‐carboxylate methyl 4‐(2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethoxy)benzoate 4‐(2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethoxy)benzoic acid 6‐(methanesulfonyl)‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐[(3R)‐3‐aminopyrrolidin‐1‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride salt N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}cyclopropanecarboxamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐2,2‐difluoroacetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐2‐methoxyacetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}oxetane‐3‐carboxamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐1‐methylazetidine‐3‐carboxamide methyl {(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}carbamate N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}methanesulfonamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}cyclopropanesulfonamide cyclopropyl{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}methanone 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2‐methoxyethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2,2‐difluoroethan‐1‐one {4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}(oxetan‐3‐yl)methanone 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2‐(dimethylamino)ethan‐1‐one {4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}(1‐fluorocyclopropyl)methanone 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2,2‐difluoropropan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazine‐1‐carbonyl}cyclopropane‐1‐carbonitrile methyl 10‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐10‐oxodecanoate 10‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐10‐oxodecanoic acid 4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐N,N‐dimethylpiperazine‐1‐carboxamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[4‐(methanesulfonyl)piperazin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine 2‐amino‐1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2‐(methylamino)ethan‐1‐one 3‐amino‐1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}propan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐3‐(methylamino)propan‐1‐one 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐methyl‐N‐{(1R)‐1‐[3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 2‐[2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]‐ 2,6‐diazaspiro[3.4]octan‐7‐one 1‐{4‐[2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one 2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐fluoro‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐ 4‐amine 2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 2‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 1‐{4‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}‐6‐(1‐oxa‐6‐azaspiro[3.3]heptan‐ 6‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine 6‐fluoro‐2,8‐dimethyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐ amine 1‐{4‐[2,8‐dimethyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one 2,8‐dimethyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2,8‐dimethyl‐N‐{(1R)‐1‐[3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 2‐[2,8‐dimethyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]‐ 2,6‐diazaspiro[3.4]octan‐7‐one 6‐fluoro‐2,8‐dimethyl‐N‐{(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 1‐{4‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 2‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2,8‐dimethyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2,8‐dimethyl‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(3R)‐1‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3S)‐1‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide 6‐chloro‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 2‐methyl‐6‐(1‐methyl‐1H‐pyrazol‐4‐yl)‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐(4,5‐dihydrofuran‐2‐yl)‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐(2,5‐dihydrofuran‐3‐yl)‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐(3,6‐dihydro‐2H‐pyran‐4‐yl)‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐(5,6‐dihydro‐2H‐pyran‐3‐yl)‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 2‐methyl‐6‐(1‐methyl‐1,2,3,6‐tetrahydropyridin‐4‐yl)‐N‐{(1R)‐1‐[3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 2‐methyl‐6‐[(3RS)‐oxolan‐3‐yl]‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine amine (mixture of stereoisomers) 2‐methyl‐6‐(oxan‐4‐yl)‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐ amine 2‐methyl‐6‐[(3RS)‐oxan‐3‐yl]‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine (mixture of stereoisomers) 2‐methyl‐6‐(1‐methylpiperidin‐4‐yl)‐N‐{(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine methyl 2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxylate 2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxamide N,2‐dimethyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxamide 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐4‐carbonitrile N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐6‐[(2S)‐2,4‐dimethylpiperazin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine {1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐4‐methylpiperazin‐2‐yl}methanol (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐ dihydroimidazo[1,2‐a]pyrazin‐7(8H)‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐ dihydro[1,2,4]triazolo[1,5‐a]pyrazin‐7(8H)‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine 6‐(cyclobutyloxy)‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine 6‐butoxy‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[2‐ (methylamino)ethoxy]pyrido[3,4‐d]pyrimidin‐4‐amine N‐[(1R)‐1‐{3‐(difluoromethyl)‐2‐[2‐(methylamino)ethoxy]phenyl}ethyl]‐2‐methyl‐6‐[2‐ (methylamino)ethoxy]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐[(oxetan‐3‐yl)oxy]pyrido[3,4‐ d]pyrimidin‐4‐amine tert‐butyl 3‐{[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]oxy}azetidine‐1‐carboxylate N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐{[(3R)‐oxolan‐3‐ yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐{[(3R)‐oxolan‐3‐yl]oxy}phenyl]ethyl}‐2‐methyl‐6‐{[(3R)‐oxolan‐ 3‐yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methyl‐6‐{[(3S)‐oxolan‐3‐ yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐{[(3S)‐oxolan‐3‐yl]oxy}phenyl]ethyl}‐2‐methyl‐6‐{[(3S)‐oxolan‐ 3‐yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐{[(3S)‐1‐methylpyrrolidin‐3‐yl]oxy}phenyl]ethyl}‐2‐methyl‐6‐ {[(3S)‐1‐methylpyrrolidin‐3‐yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine 6‐[(azetidin‐3‐yl)oxy]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine hydrochloride tert‐butyl {(3‐trans)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐4‐fluoropyrrolidin‐3‐yl}carbamate (mixture of stereoisomers) 6‐[(trans)‐3‐amino‐4‐fluoropyrrolidin‐1‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride (mixture of stereoisomers) tert‐butyl {(cis)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐4‐fluoropyrrolidin‐3‐yl}carbamate (mixture of stereoisomers) 6‐[(cis)‐3‐amino‐4‐fluoropyrrolidin‐1‐yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride (mixture of stereoisomers) or a stereoisomer, a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. 12. A SOS1 inhibitor compound as described herein or in claim 1 for use in the treatment and/or prevention of cancer, wherein said SOS1 inhibitor compound is administered in combination with at least one other pharmacologically active substance and wherein each of said other pharmacologically active substance(s) is selected from the group consisting of: an inhibitor of HRas, NRas or KRAS and mutants thereof, in particular an inhibitor of KRAS‐G12C; an inhibitor of MAP kinases, in particular MEK1, MEK2, ERK1, ERK2, ERK5 and/or of an inhibitor of PI3‐ kinases and mutants thereof; an inhibitor of Tropomyosin Receptor kinases and/or of mutants thereof; an inhibitor of SHP2 and mutants thereof; inhibitor of EGFR and/or of mutants thereof; an inhibitor of FGFR1 and/or FGFR2 and/or FGFR3 and/or of mutants thereof; an inhibitor of ALK and/or of mutants thereof; an inhibitor of c‐MET and/or of mutants thereof; an inhibitor of BCR‐ABL and/or of mutants thereof; an inhibitor of ErbB2 (Her2) and/or of mutants thereof; an inhibitor of AXL and/or of mutants thereof; an inhibitor of A‐Raf and/or B‐ Raf and/or C‐Raf and/or of mutants thereof; an inhibitor of mTOR and mutants thereof; an inhibitor of IGF1/2 and/or of IGF1‐R; an inhibitor of farnesyl transferase. |
The present invention covers 2-methyl-aza-quinazoline compounds of general formula (I) as described and defined herein, methods of preparing said compounds, intermediate compounds useful for preparing said compounds, pharmaceutical compositions and combinations comprising said compounds, and the use of said compounds for manufacturing pharmaceutical compositions for the treatment or prophylaxis of diseases, in particular of hyperproliferative disorders, as a sole agent or in combination with other active ingredients.
BACKGROUND
The present invention covers 2-methyl-aza-quinazoline compounds of general formula (I) which inhibit the Ras-Sosl interaction.
US 2011/0054173 A1 discloses certain 1- or 2-(4-(aryloxy)-phenyl)ethylamino-, oxy- or sulfanyl)pteridines and 1- or 2-(4-(heteroaryloxy)-phenyl)ethylamino-, oxy- or sulfanyl)pteridines and their use as agrochemicals and animal health products.
In the 2-position substituted quinazoline compounds are described e.g. in EP 0326328, EP 0326329, W093/007124, W02003/087098 and US 5,236,925. These compounds are either not described as pharmaceutically active compounds or, if they are described as pharmacologically active compounds, they are described as compounds having affinity to the Epidermal Growth Factor Receptor (EGFR).
In the majority (45-100%) of patients receiving EGFR inhibitors skin toxicity is a class-specific side effect that is typically manifested as a papulopustular rash. The skin toxicity is related to the inhibition of EGFR in the skin, which is crucial for the normal development and physiology of the epidermis.
Flowever, the state of the art does not describe: the 2-methyl substituted quinazoline compounds of general formula (I) of the present invention as described and defined herein, i.e. compounds having a quinazoline core bearing a methyl group on the carbon atom 2 which effectively and selectively inhibit the Ras-Sosl interaction without significantly targeting the EGFR receptor.
Ras proteins play an important role in human cancer. Mutations in Ras proteins can be found in 20- 30% of all human tumors and are recognized as tumorigenic drivers especially in lung, colorectal and pancreatic cancers (Malumbres & Barbacid 2002 Nature Reviews Cancer, Pylayeva-Gupta et al. 2011 Nature Reviews Cancer). Three human Ras genes are known that encode four different Ras proteins of 21 kDa size: Fl-Ras, N-Ras, and two splice variants of K-Ras, namely K-Ras 4A and K-Ras- 4B. All Ras isoforms are highly conserved within the GTP-binding domain and differ mainly in the hypervariable C-terminal region. The C-termini of the different Ras-isoforms are posttranslationally modified by lipidation (farnesylation, palmitoylation) to facilitate membrane anchorage. The localization of Ras-proteins at the cytoplasmic membrane provides vicinity to transmembrane growth receptors and has been shown to be essential for transmitting growth signals from extracellular growth factor binding to intracellular downstream pathways. A variety of upstream signals may activate Ras proteins depending on the cellular context, such as epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), nerve growth factor receptor (NGFR) and others. Activated Ras can signal through various downstream pathways, e.g. the Raf-MEK-ERK or the PI3K-PDK1-Akt pathways.
On the molecular level, Ras proteins function as molecular switches. By binding GTP and GDP they exist in an active (GTP-bound) and inactive (GDP-bound) state in the cell. Active GTP-loaded Ras recruits other proteins by binding of their cognate Ras-binding domains (RBDs) resulting in activation of the effector protein followed by downstream signalling events of diverse functions, e.g. cytoskeletal rearrangements or transcriptional activation. The activity status of Ras is tightly regulated by guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs). GEFs function as activators of Ras by promoting the nucleotide exchange from GDP to GTP. GAPs deactivate Ras-GTP by catalyzing the hydrolysis of the bound GTP to GDP. In a cancer cell, point mutations, typically within the GTP-binding region at codon 12, eliminate the ability of RAS to efficiently hydrolyse bound GTP, even in the presence of a GAP. Therefore, cancer cells comprise increased levels of active mutated Ras-GTP, which is thought to be a key factor for driving cancer cell proliferation.
Three main families of RAS-specific GEFs have been identified so far (reviewed in Vigil 2010 Nature Reviews Cancer; Rojas et al 2011, Genes & Cancer 2(3) 298-305). There are two son of sevenless proteins (SOS1 and SOS2), 4 different isoforms of Ras guanine nucleotide releasing proteins (Ras- GRP1-4) and two Ras guanine nucleotide releasing factors (Ras-GRFl and 2). The SOS proteins are ubiquitously expressed and are recruited to sites of activated growth factors. Ras-GRFs are expressed mainly in the nervous system, where they are involved in Calcium-dependent activation of Ras. In contrast, Ras GRP proteins are expressed in hematopoietic cells and act in concert with non-receptor tyrosine kinases. In the context of cancer, mainly SOS proteins have been found to be involved.
Targeting Ras for cancer therapy has been a dream since the 1990s (Downward 2002 Nature Reviews Cancer, Krens et al. 2010 Drug Discovery Today). Due to the compact nature, the high affinity towards GDP and GTP in combination with high intracellular GTP concentrations, the Ras protein itself has always been considered to be undruggable, i.e. the chance to identify small chemical molecules that would bind to and inhibit active Ras was rated extremely low. Alternative approaches have been undertaken to reduce Ras signaling, e.g. by addressing more promising drug targets such as enzymes involved in the posttranslational modification of Ras proteins, especially farnesyltransferase and geranylgeranyltransferase (Berndt 2011 Nature Reviews Cancer). Inhibitors of farnesyltransferase (FTIs) were identified and developed with promising antitumor effects in preclinical models. Unexpectedly, in clinical trials these inhibitors have been of limited efficacy. Targeting upstream and downstream kinases involved in Ras signaling pathways has been more successful. Several drugs are and have been in clinical trials that inhibit different kinases, e.g. EGFR, Raf, MEK, Akt, PI3K (Takashima & Falter 2013 Expert Opin. Ther. Targets). Marketed cancer drugs are available that inhibit Raf, EGFR or MEK.
Nevertheless, there is still a large unmet need for the treatment of Ras-dependent tumors that are resistant against current therapies. Many research groups have been active to identify small molecules that target Ras directly (Ras small molecules have been reviewed in: Cox et al. 2014 Nature Reviews Drug Discovery, Spiegel et al. 2014 Nature Chemical Biology, Cromm 2015 Angewandte Chemie, Marin-Ramos et al Seminars in Cancer Biology). One group of inhibitors comprises small molecules that inhibit the interaction of Ras with its effectors Raf or PI3K. Another group of compounds acts as covalent inhibitors of a specific cysteine mutant form of K-Ras (glycine to cysteine point mutation G12C). The specific targeting of the Ras-G12C mutant might have the benefit of reduced side effects, as the wildtype Ras proteins should not be affected. Furthermore, several reports show small molecules and peptides that interrupt the GEF assisted activation of Ras (Hillig et al 2019 PNAS; Gray et al 2019 Angewandte Chemie). There seem to be several different binding sites possible that result in this mode of action. Inhibitors may bind to Ras or to the GEF in an allosteric or orthosteric fashion. All these approaches of direct Ras-targeting are in preclinical research stage. Stabilized peptides have been shown to be active in the nanomolar range. (Leshchiner et al. 2015 PNAS). Their usefulness as drugs in a clinical setting has to be awaited.
The Epidermal Growth Factor Receptor (EGFR) is a tyrosine kinase (TK) receptor that is activated upon binding to the Epidermal Growth Factor and other growth factor ligands, triggering several downstream pathways, including RAS/MAPK, PI3K/Akt and STAT that regulate different cellular processes, including DNA synthesis and proliferation (Russo A, Oncotarget.4254, 2015). The family of HER (ErbB) receptor tyrosine kinases consists of four members, ie, epidermal growth factor receptors [EGFR (FIERI or ErbBl), HER2 (ErbB2, neu), HER3 (ErbB3), and HER4 (ErbB4)]. Overexpression, mutation, or aberrant activity of these receptors has been implicated in various types of cancer (Feldinger K, Breast Cancer (Dove Med Press), 2015, 7, 147).
First-generation inhibitors Erlotinib and Gefitinib are small molecule inhibitors of the EGFR/HER‐1 (human epidermal growth factor receptor) tyrosine kinase. Erlotinib and Gefitinib were developed as reversible and highly specific small‐molecule tyrosine kinase inhibitors that competitively block the binding of ad enosine triphosphate to its binding site in the tyrosine kinase domain of EGFR, thereby inhibiting autophosphorylation and blocking downstream signaling ( Cataldo VD, N Engl J Med, 2011, 364, 947). Second‐generation inhibitors Afatinib is an oral tyrosine kinase inhibitor (TKI) approved for the first‐line treatment of patients w ith NSCLC whose tumors are driven by activating mutations of genes coding for epidermal growth factor receptor (EGFR). Afatinib is also an inhibitor of a specific EGFR mutation (T790M) that causes resistance to first‐generation EGFR‐targeted TKIs in about half of patients receiving those drugs. (Engle JA, Am J Health Syst Pharm 2014, 71 (22), 1 933). Neratinib, a pan‐HER inhibitor, irreversible tyrosine kinase inhibitor binds and inhibits the tyrosine kinase activity of epidermal growth factor receptors, EGFR (or HER1), HER2 and HER4, which leads to reduced phosphorylation and activation of downstream signaling pathways. Neratinib has been shown to be effective against HER2‐overexpressing or mutant tumors in vitro and in vivo. Neratinib is currently being investigated in various clinical trials in breast cancers and other solid tumors, including those with HER2 mutation (Feldinger K, Brea st Cancer (Dove Med Press), 2015, 7, 147). Dacomitinib is an irreversible inhibitor of EGFR, HER2, and HER4. In preclinical cell lines and xenograft studies, dacomitinib demonstrated activities against both activating EGFR mutations and EGFR T790M (Liao BC, Curr Opin Oncol. 2015, 27(2), 94). Third‐generation inhibitors The third‐generation EGFR‐TKIs were designed to in hibit EGFR T790M while sparing wild‐type EGFR. AZD9291 (AstraZeneca, Macclesfield, UK), a mono‐anilino‐pyrimidine compound, is an irreversible mutant selective EGFR‐TKI. This drug is structurally different from the first and second‐generation EGFR‐TKIs. In preclinical studies, it potently inhibited phosphorylation of EGFR in cell lines with activating EGFR mutations (EGFR del19 and EGFR L858R) and EGFR T790M. AZD9291 also caused profound and sustained tumor regression in tumor xenograft and transgenic mouse models harboring activating EGFR mutations and EGFR T790M. AZD9291 was less potent in inhibiting phosphorylation of wild‐type EGFR cell lines (Liao BC, Curr Opin Oncol. 2015, 27(2), 94). Rociletinib (CO‐1686) (Clovis Oncology, Boulder, Colo ), a 2,4‐disubstituted pyrimidine molecule, is an irreversible mutant selective EGFR‐TKI. In preclinica l studies, CO‐1686 led to tumor regression in cell ‐ lines, xenograft models, and transgenic mouse models harboring activating EGFR mutations and EGFR T790M (Walter AO, Cancer Discov, 2013, 3(12), 1 404). HM61713 (Hanmi Pharmaceutical Company Ltd, Seoul, South Korea) is an orally administered, selective inhibitor for activating EGFR mutations and EGFR T790M. It has low activity against wild‐ type EGFR (Steuer CE, Cancer. 2015, 121(8), E1). Hillig et al 2019 PNAS describe compounds like as a potent SOS1 inhibitor and as a tool compound for further investigation of RAS‐SOS1 biology in vitro. WO2018/172250 (Bayer Pharma AG) describes 2‐methyl‐ quinazoline like as inhibiting Ras‐Sos interaction. WO 2018/115380 (Boehringer Ingelheim) describes benzyla mino substituted quinazolines like as SOS1 inhibitors. WO2019/122129 (Boehringer Ingelheim) describes benzylami nosubstituted pyridopyrimidinoes like
as SOS1 inhibitors. It has now been found, and this constitutes the bas is of the present invention, that the compounds of the present invention have surprising and advantag eous properties. In particular, the compounds of the present invention have surprisingly been found to effectively and selectively inhibit the Ras‐Sos1 interaction without significantly targeting the EGFR receptor and may therefore be used for the treatment or prophylaxis of hyper‐proliferative disorders, in particular cancer. Furthermore the compounds of the present invention sh ow good metablic stability and permeability. DESCRIPTION OF THE INVENTION In accordance with a first aspect, the present inven tion covers compounds of general formula (I): wherein R 1 is selected from ‐H, halogen, ‐OH, ‐CN, ‐NO 2 , C 1 ‐C 6 ‐alkylsulfanyl, ‐NR a R b , wherein R a and R b are independently selected from ‐H or C 1 ‐C 6 ‐alkyl, C 1 ‐C 6 ‐alkyl, C 1 ‐C 6 ‐alkoxy, C 2 ‐C 6 ‐alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl, C 4 ‐C 8 ‐cycloalkenyl, 4‐ to 7‐membered heterocycloalkyl, 5‐ to 10 membered heterocycloalkenyl, heterospirocycloalkyl, fused heterocycloalkyl, bridged heterocycloalkyl, phenyl, heteroaryl, C 1 ‐C 6 ‐haloalkyl, ‐C(=O)OH, ‐C(=O)OR c , wherein R c stands for C 1 ‐C 6 ‐alkyl, C 3 ‐C 6 ‐alkenyl, C 3 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl or C 4 ‐C 8 ‐cycloalkenyl, ‐N=S(=O)(R d )R e , wherein R d and R e are independently selected from C 1 ‐C 6 ‐alkyl, C 2 ‐C 6 ‐ alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl or C 4 ‐C 8 ‐cycloalkenyl, ‐NH‐C(O)‐C 1 ‐C 6 ‐alkyl, ‐NH‐C(O)‐NR a R b , wherein R a and R b are selected independently from a hydrogen ato m or a C 1 ‐C 6 ‐alkyl, ‐NH‐(CH 2 ) k ‐NH‐C(O)‐C 1 ‐C 6 ‐alkyl, wherein k is 1 or 2, ‐NH‐(CH 2 ) l ‐R f , wherein l is 0, 1 or 2 and R f stands for a 4‐ to 7‐membered heterocycloalkyl, heteroaryl or C 1 ‐C 6 ‐alkylsulfonyl, whereby in all foregoing definitions the C 1 ‐C 6 ‐alkyl‐, C 1 ‐C 6 ‐alkoxy‐, the 4‐ to 7‐ membered heterocycloalkyl and the heteroaryl can be o ptionally substituted, one or two or three times, identically or differently, w ith a halogen atom, hydroxy, oxo (=O), a cyano, nitro, C 1 ‐C 6 ‐alkyl, C 2 ‐C 6 ‐alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl, 4‐ to 7‐membered heterocycloalkyl, C 1 ‐C 6 ‐alkoxy, C 1 ‐C 6 ‐haloalkyl, C 1 ‐C 6 ‐haloalkoxy, C 1 ‐ C 6 ‐alkylsulfonyl, phenyl, benzyl, heteroaryl, ‐CH 2 ‐heteroaryl, C 3 ‐C 8 ‐cycloalkoxy. phenyloxy, heteroaryloxy, ‐NH‐C(O)‐C 1 ‐C 6 ‐alkyl or –NR a R b , wherein R a and R b are independently selected from a hydrogen atom or C 1 ‐C 6 ‐alkyl, ‐O‐(CH 2 ) z ‐phenyl, ‐O(CH 2 ) z ‐C 4 ‐C 7 ‐heterocycloalkyl, ‐O(CH 2 ) z ‐heteroaryl, wherein z is 0, 1 or 2, and the phenyl, heterocycloalkyl and heteroaryl can optionally be substituted with a group selected from hydroxy, heterocycloalkyl or he terocaclyoalkenyl, which both can be substituted with a methyl‐ and/or oxo‐group, wherein L 2 a stands for C(O), L 2 b stands for a bond or C 1 ‐C 6 ‐ alkylene, X2 stands for , and Rx 2 stands for or in which a further R 1 as defined above can be directly attached to a first R 1 equaling C 1 ‐C 6 ‐alkyl, C 1 ‐C 6 ‐alkoxy, C 2 ‐C 6 ‐alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl, C 4 ‐C 8 ‐cycloalkenyl, 4‐ to 7‐membered heterocycloalkyl, 5‐ to 10 membered heterocycloalkenyl, heterospirocycloalkyl, fused heterocycloalkyl, bridged heterocycloalkyl, phenyl, heteroaryl, C 1 ‐C 6 ‐haloalkyl, y is 1, 2 or 3; and either both T and V stand for nitrogen or T s tands for carbon and V for nitrogen or T for nitrogen and V for carbon; A is selected from the group consisting of C 6‐10 aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; R 2 is each independently selected from the group consisting of C 1‐4 alkyl, C 2‐4 alkenyl, C 2‐ 4 alkinyl, C 1‐4 haloalkyl, hydroxy‐C 1‐4 alkyl, hydroxy‐C 1‐4 haloalkyl, C 3‐6 cycloalkyl, 3‐6 membered heterocyclyl, hydroxy‐C 3‐6 cycloalkyl, C 1‐4 haloalkyl substituted with a 3‐6 membered heterocyclyl, 3‐6 membered heterocyclyl substituted with hydroxy, halogen, ‐NH 2 , ‐SO 2 ‐C 1‐4 alkyl and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring; x is 1, 2 or 3; or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. In accordance with a further first aspect, the prese nt invention covers compounds of general formula (Ia): wherein R 1 is selected from ‐H, halogen, ‐OH, ‐CN, ‐NO 2 , C 1 ‐C 6 ‐alkylsulfanyl, ‐NR a R b , wherein R a and R b are independently selected from ‐H or C 1 ‐C 6 ‐alkyl, C 1 ‐C 6 ‐alkyl, C 1 ‐C 6 ‐alkoxy, C 2 ‐C 6 ‐alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl, C 4 ‐C 8 ‐cycloalkenyl, 4‐ to 7‐membered heterocycloalkyl, 5‐ to 10 membered heterocycloalkenyl, heterospirocycloalkyl optionally substituted by an oxo‐group (=O), fused heterocycloalkyl optionally substituted by an oxo‐group (=O), bridged heterocycloalkyl optionally substituted by an oxo‐gro up (=O), phenyl, heteroaryl, C 1 ‐C 6 ‐ haloalkyl, ‐C(=O)OH, ‐C(=O)OR c , wherein R c stands for C 1 ‐C 6 ‐alkyl, C 3 ‐C 6 ‐alkenyl, C 3 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl or C 4 ‐C 8 ‐cycloalkenyl, ‐N=S(=O)(R d )R e , wherein R d and R e are independently selected from C 1 ‐C 6 ‐alkyl, C 2 ‐C 6 ‐ alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl or C 4 ‐C 8 ‐cycloalkenyl, ‐NH‐C(O)‐C 1 ‐C 6 ‐alkyl, ‐NH‐C(O)‐NR a R b , wherein R a and R b are selected independently from a hydrogen ato m or a C 1 ‐C 6 ‐alkyl, ‐NH‐(CH 2 ) k ‐NH‐C(O)‐C 1 ‐C 6 ‐alkyl, wherein k is 1 or 2, ‐NH‐(CH 2 ) l ‐R f , wherein l is 0, 1 or 2 and R f stands for a 4‐ to 7‐membered heterocycloalkyl, heteroaryl or C 1 ‐C 6 ‐alkylsulfonyl, whereby in all foregoing definitions the C 1 ‐C 6 ‐alkyl‐, C 1 ‐C 6 ‐alkoxy‐, the 4‐ to 7‐ membered heterocycloalkyl and the heteroaryl can be o ptionally substituted, one or two or three times, identically or differently, w ith a halogen atom, hydroxy, oxo (=O), a cyano, nitro, C 1 ‐C 6 ‐alkyl, C 2 ‐C 6 ‐alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl, 4‐ to 7‐membered heterocycloalkyl, C 1 ‐C 6 ‐alkoxy, C 1 ‐C 6 ‐haloalkyl, C 1 ‐C 6 ‐haloalkoxy, C 1 ‐ C 6 ‐alkylsulfonyl, phenyl, benzyl, heteroaryl, ‐CH 2 ‐heteroaryl, C 3 ‐C 8 ‐cycloalkoxy. phenyloxy, heteroaryloxy, ‐NH‐C(O)‐C 1 ‐C 6 ‐alkyl or –NR a R b , wherein R a and R b are independently selected from a hydrogen atom or C 1 ‐C 6 ‐alkyl, ‐O‐(CH 2 ) z ‐phenyl, ‐O(CH 2 ) z ‐C 4 ‐C 7 ‐heterocycloalkyl, ‐O(CH 2 ) z ‐heteroaryl, wherein z is 0, 1 or 2, and the phenyl, heterocycloalkyl and heteroaryl can optionally be substituted with a group selected from hydroxy, heterocycloalkyl or he terocaclyoalkenyl, which both can be substituted with a methyl‐ and/or oxo‐group, wherein L 2 a stands for C(O), L 2 b stands for a bond or C 1 ‐C 6 ‐ alkylene, X2 stands for and Rx 2 stands for or in which a further R 1 as defined above can be directly attached to a first R 1 equaling C 1 ‐C 6 ‐alkyl, C 1 ‐C 6 ‐alkoxy, C 2 ‐C 6 ‐alkenyl, C 2 ‐C 6 ‐alkynyl, C 3 ‐C 8 ‐cycloalkyl, C 4 ‐C 8 ‐cycloalkenyl, 4‐ to 7‐membered heterocycloalkyl, 5‐ to 10 membered heterocycloalkenyl, heterospirocycloalkyl, fused heterocycloalkyl, bridged heterocycloalkyl, phenyl, heteroaryl, C 1 ‐C 6 ‐haloalkyl, y is 1, 2 or 3; and either both T and V stand for nitrogen or T s tands for carbon and V for nitrogen or T for nitrogen and V for carbon; A is selected from the group consisting of C 6‐10 aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; R 2 is each independently selected from the group consisting of C 1‐4 alkyl, C 2‐4 alkenyl, C 2‐ 4 alkinyl, C 1‐4 haloalkyl, hydroxy‐C 1‐4 alkyl, hydroxy‐C 1‐4 haloalkyl, C 3‐6 cycloalkyl, 3‐6 membered heterocyclyl, hydroxy‐C 3‐6 cycloalkyl, C 1‐4 haloalkyl substituted with a 3‐6 membered heterocyclyl, 3‐6 membered heterocyclyl substituted with hydroxy, halogen, ‐NH 2 , ‐SO 2 ‐C 1‐4 alkyl and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring; R 6 is selected from the group consisting of ‐H, halog en, C 1 ‐ 4 alkyl, C 3‐7 ‐cycloalkyl, C 4‐ 7 heterocycloalkyl optionally comprising 1 or 2 ni trogen, 1 oxygen or 1 sulphur atom, ‐ O‐C 1‐4 alkyl, ‐NH 2 , ‐NH(C 1‐4 alykl) or ‐NH(C 1‐4 alkyl) 2 , x is 1, 2 or 3; or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. Alternatively R 6 of formula (Ia) is selected from the group c onsisting of ‐H, ‐CH 3 , ‐CH(CH 3 ) 2 , ‐CH 2 OH, ‐ CF 3 or ‐CHF 2 . DEFINITIONS When groups in the compounds according to the invention are substituted, it is possible for said groups to be mono‐substituted or poly‐substituted with substituent(s), unless otherwise specified. Within the scope of the present invention, the meanings of all groups which occur repeatedly are independent from one another. It is possible that groups in the compounds according to the invention are substituted with one, two or three ide ntical or different substituents, particularly with one substituent. As used herein, an oxo substituent represents an oxy gen atom, which is bound to a carbon atom or to a sulfur atom via a double bond. The term “ring substituent” means a substituent a ttached to an aromatic or nonaromatic ring which replaces an available hydrogen atom on the ring. Should a composite substituent be composed of more than one parts, e.g. (C 1 ‐C 4 ‐alkoxy)‐(C 1 ‐C 4 ‐alkyl)‐, it is possible for the position o f a given part to be at any suitable position of said composite substituent, i.e. the C 1 ‐C 4 ‐alkoxy part can be attached to any carbon a tom of the C 1 ‐C 4 ‐alkyl part of said (C 1 ‐C 4 ‐alkoxy)‐(C 1 ‐C 4 ‐alkyl)‐ group. A hyphen at the beginning o r at the end of such a composite substituent indicates the point of attachment of said composite substituent to the rest of the molecule. Should a ring, comprising carbon atoms and optionally one or more heteroatoms, such as nitrogen, oxygen or sulfur atoms for example, be substituted with a substituent, it is possible for said substituent to be bound at any suitable position of said ring, be it bound to a suitable carbon atom and/or to a suitabl e heteroatom. The term “comprising” when used in the specificat ion includes “consisting of”. If within the present text any item is referred to as “as mentioned herein”, it means that it may be mentioned anywhere in the present text. The terms as mentioned in the present text have the following meanings: The term “halogen atom” means a fluorine, chlorin e, bromine or iodine atom, particularly a fluorine, chlorine or bromine atom. The term “C 1 ‐C 6 ‐alkyl” means a linear or branched, saturate d, monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, e.g. a methyl, ethyl, propyl, isopropyl, butyl, sec‐butyl, isobutyl, tert‐ butyl, pentyl, isopentyl, 2‐methylbutyl, 1‐methylbut yl, 1‐ethylpropyl, 1,2‐dimethylpropyl, neo‐pentyl, 1,1‐dimethylpropyl, hexyl, 1‐methylpentyl, 2‐methylpentyl, 3‐methylpentyl, 4‐methylpentyl, 1‐ethylbutyl, 2‐ethylbutyl, 1,1‐dimethylbutyl, 2,2‐dimethylbutyl, 3,3‐dimethylbutyl, 2,3‐dimethylbutyl, 1,2‐dimethylbutyl or 1,3‐dimethy lbutyl group, or an isomer thereof. Particularly, said group has 1, 2, 3 or 4 carbon atoms (“C 1 ‐C 4 ‐alkyl”), e.g. a methyl, ethyl, propyl, isop ropyl, butyl, sec‐butyl isobutyl, or tert‐butyl group, more part icularly 1, 2 or 3 carbon atoms (“C 1 ‐C 3 ‐alkyl”), e.g. a methyl, ethyl, n‐propyl or isopropyl group. The term “C 1 ‐C 6 ‐hydroxyalkyl” means a linear or branched, saturated, monovalent hydrocarbon group in which the term “C 1 ‐C 6 ‐alkyl” is defined supra, and in which 1, 2 or 3 hydrogen atoms are replaced with a hydroxy group, e.g. a hydroxymethyl, 1‐hydroxyethyl, 2‐hydroxyethyl, 1,2‐dihydroxyethyl, 3‐hydroxypropyl, 2‐hydroxypropyl, 1‐hydroxypropyl, 1‐hydroxypropan‐2‐yl, 2‐hydroxypropan‐2‐yl, 2,3‐dihydroxypropyl, 1,3‐dihydroxypropan‐2‐yl, 3‐hydroxy‐2‐methyl‐propyl, 2‐hydroxy‐2‐methyl‐propyl, 1‐hydroxy‐2‐methyl ‐propyl group. The term “C 1 ‐C 6 ‐alkylsulfanyl” means a linear or branched, saturated, monovalent group of fo rmula (C 1 ‐C 6 ‐alkyl)‐S‐, in which the term “C 1 ‐C 6 ‐alkyl” is as defined supra, e.g. a methylsulfanyl, ethylsulfanyl, propylsulfanyl, isopropylsulfanyl, butylsu lfanyl, sec‐butylsulfanyl, isobutylsulfanyl, tert‐ butylsulfanyl, pentylsulfanyl, isopentylsulfanyl, hexylsu lfanyl group. The term “C 1 ‐C 6 ‐alkylsulfonyl” means a linear or branched, saturated, monovalent group of fo rmula (C 1 ‐C 6 ‐alkyl)‐SO 2 ‐, in which the term “C 1 ‐C 6 ‐alkyl” is as defined supra, e.g. a methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsu lfonyl, sec‐butylsulfonyl, isobutylsulfonyl, tert‐ butylsulfonyl, pentylsulfonyl, isopentylsulfonyl, hexylsu lfonyl group. The term “C 1 ‐C 6 ‐alkoxy” means a linear or branched, saturated, monovalent group of formula (C 1 ‐C 6 ‐alkyl)‐O‐, in which the term “C 1 ‐C 6 ‐alkyl” is as defined supra, e.g. a methoxy, ethoxy, n‐propoxy, isopropoxy, n‐butoxy, sec‐butoxy, isobutoxy, tert‐butoxy, pentyloxy, isopentyloxy or n‐hexyloxy group, or an isomer thereof. The term “C 2 ‐C 6 ‐alkenyl” means a linear or branched, monovalent hydrocarbon group, which contains one or two double bonds, and which has 2, 3, 4, 5 or 6 carbon atoms, particularly 2 or 3 carbon atoms (“C 2 ‐C 3 ‐alkenyl”), it being understood that in the case in which said alkenyl group contains more than one double bond, then it is poss ible for said double bonds to be isolated from, or conjugated with, each other. Said alkenyl group is, for example, an ethenyl (or “vinyl”), prop‐2‐en‐1‐yl (or “allyl”), prop‐1‐en‐1‐yl, but‐3‐enyl, but‐2‐enyl, but‐1‐enyl, pent‐4‐enyl, pent‐3‐enyl, pent‐2‐enyl, pent‐1‐enyl, hex‐5‐enyl, hex‐4‐enyl, hex‐3‐enyl, hex‐2‐enyl, hex‐1‐enyl, prop‐1‐en‐2‐yl (or “isopropenyl”), 2‐methylprop‐2‐enyl, 1‐methylprop‐2‐enyl, 2‐methylprop‐1‐enyl, 1‐methylprop‐1‐enyl, 3‐methylbut‐3‐enyl, 2‐methylbut‐3‐enyl, 1‐methylbut‐3‐enyl, 3‐methylbut‐2‐enyl, 2‐methylbut‐2‐enyl, 1‐methylbut‐2‐enyl, 3‐methylbut‐1‐enyl, 2‐methylbut‐1‐enyl, 1‐methylbut‐1‐enyl, 1,1‐ dimethylprop‐2‐enyl, 1‐ethylprop‐1‐enyl, 1‐pro pylvinyl, 1‐isopropylvinyl, 4‐methylpent‐4‐enyl, 3‐methylpent‐4‐enyl, 2‐methylpent‐4‐enyl, 1‐methylpent‐4‐enyl, 4‐methylpent‐3‐enyl, 3‐methylpent‐3‐enyl, 2‐methylpent‐3‐enyl, 1‐methylpent‐3‐enyl, 4‐methylpent‐2‐enyl, 3‐methylpent‐2‐enyl, 2‐methylpent‐2‐enyl, 1‐methylpent‐2‐enyl, 4‐methylpent‐1‐enyl, 3‐methylpent‐1‐enyl, 2‐methylpent‐1‐enyl, 1‐methylpent‐1‐enyl, 3‐ethylbut‐3‐enyl, 2‐ethylbut‐3‐enyl, 1‐ethylbut‐3‐enyl, 3‐ethylbut‐2‐enyl, 2‐ethylbut‐2‐enyl, 1‐ethylbut‐2‐enyl, 3‐ethylbut‐1‐enyl, 2‐ethylbut‐1‐enyl, 1‐ethylbut‐1‐enyl, 2‐propylprop‐2‐enyl, 1‐propylprop‐2‐enyl, 2‐isopropylprop‐2‐enyl, 1‐isopropylprop‐2‐enyl, 2‐propylprop‐1‐enyl, 1‐propylprop‐1‐enyl, 2‐isopropylprop‐1‐enyl, 1‐isopropylprop‐1‐enyl, 3,3‐dimethylprop‐1‐enyl, 1‐(1,1‐dimethylethyl)ethenyl, buta‐1,3‐dienyl, penta‐1,4‐dienyl or hexa‐1,5‐dienyl group. Particularly, said group is vinyl or allyl. The term “C 2 ‐C 6 ‐alkynyl” means a linear or branched, monova lent hydrocarbon group which contains one triple bond, and which contains 2, 3, 4, 5 or 6 carbon atoms, particularly 2 or 3 carbon atoms (“C 2 ‐C 3 ‐alkynyl”). Said C 2 ‐C 6 ‐alkynyl group is, for example, ethynyl, prop‐1‐ynyl, prop‐2‐ynyl (or “propargyl”), but‐1‐ynyl, but‐2‐ynyl, but‐3‐ynyl, pent‐1‐ynyl, pent‐2‐ynyl, pent‐3‐ynyl, pent‐4‐ynyl, hex‐1‐ynyl, hex‐2‐ynyl, hex‐3‐ynyl, hex‐4 ynyl, hex‐5‐ynyl, 1‐methylprop‐2‐ynyl, 2‐met hylbut‐3‐ynyl, 1‐methylbut‐3‐ynyl, 1‐methylbut‐2‐ynyl, 3‐methylbut‐1‐ynyl, 1‐ethylprop‐2‐ynyl, 3‐methylpent‐4‐ynyl, 2‐methylpent‐4‐ynyl, 1‐methylpent‐4‐ynyl, 2‐methylpent‐3‐ynyl, 1‐methylpent‐3‐ynyl, 4‐methylpent‐2‐ynyl, 1‐methylpent‐2‐ynyl, 4‐methylpent‐1‐ynyl, 3‐methylpent‐1‐ynyl, 2‐ethylbut‐3‐ynyl, 1‐ethylbut‐3‐ynyl, 1‐ethylbut‐2‐ynyl, 1‐propylprop‐2‐ynyl, 1‐isopropylprop‐2‐ynyl, 2,2‐dimethylbut‐3‐ynyl, 1,1‐dimethylbut‐3‐ynyl, 1,1‐dimethylbut‐2‐ynyl or 3,3‐dimethylbut‐1‐ynyl group. Particularly, said a lkynyl group is ethynyl, prop‐1‐ynyl or prop‐2 ynyl. The term “C 3 ‐C 8 ‐cycloalkyl” means a saturated, monovalent, mono‐ or bicyclic hydrocarbon ring which contains 3, 4, 5, 6, 7 or 8 carbon atoms ( C 3 ‐C 8 ‐cycloalkyl”). Said C 3 ‐C 8 ‐cycloalkyl group is for example, a monocyclic hydrocarbon ring, e.g. a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl group, or a bicyclic hydrocarbon ring, e.g. a bicyclo[4.2.0]octyl or octahydropentalenyl. The term “C 4 ‐C 8 ‐cycloalkenyl” means a monovalent, mono‐ or bicyclic hydrocarbon ring which contains 4, 5, 6, 7 or 8 carbon atoms and one dou ble bond. Particularly, said ring contains 4, 5 or 6 carbon atoms (“C 4 ‐C 6 ‐cycloalkenyl”). Said C 4 ‐C 8 ‐cycloalkenyl group is for example, a monocyclic hydrocarbon ring, e.g. a cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl or cyclooctenyl group, or a bicyclic hydrocarbon ring, e.g. a bicycl o[2.2.1]hept‐2‐enyl or bicyclo[2.2.2]oct‐2‐enyl. The term “C 3 ‐C 8 ‐cycloalkoxy” means a saturated, monovalent, mono‐ or bicyclic group of formula (C 3 ‐C 8 ‐cycloalkyl)‐O‐, which contains 3, 4, 5, 6, 7 or 8 carbon atoms, in which the term “C 3 ‐C 8 ‐cycloalkyl” is defined supra, e.g. a cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy or cyclooctyloxy group. The term "spirocycloalkyl" means a saturated, monovale nt bicyclic hydrocarbon group in which the two rings share one common ring carbon atom, and wherein said bicyclic hydrocarbon group contains 5, 6, 7, 8, 9, 10 or 11 carbon atoms, it being possible for said spirocycloalkyl group to be attached to the rest of the molecule via any one o f the carbon atoms except the spiro carbon atom. Said spirocycloalkyl group is, for example, spiro[2.2]pentyl, spiro[2.3]hexyl, spiro[2.4]heptyl, spiro[2.5]octyl, spiro[2.6]nonyl, spiro[3.3]heptyl, spiro[3.4]octyl, spiro[3.5]nonyl, spiro[3.6]decyl, spiro[4.4]nonyl, spiro[4.5]decyl, spiro[4.6]undecyl or s piro[5.5]undecyl. The terms “4‐ to 7‐membered heterocycloalkyl” means a monocyclic, saturated heterocycle with 4, 5, 6 or 7 ring atoms in total, which contains one or two identical or different ring heteroatoms from the series N, O and S, it being possible for said heterocycloalkyl group to be attached to the rest of the molecule via any one of the carbon atoms or, i f present, a nitrogen atom. Said heterocycloalkyl group, without being limited thereto, can be a 4‐membered ring, such as azetidinyl, oxetanyl or thietanyl, for example; or a 5‐membered ring, such as tetrahydrofuranyl, 1,3‐ dioxolanyl, thiolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, 1,1‐dioxidothiolanyl, 1, 2‐oxazolidinyl, 1,3‐oxazolidinyl or 1,3‐thiazolidinyl, for example; or a 6‐membered ring, such as tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, dithiany l, thiomorpholinyl, piperazinyl, 1,3‐dioxanyl, 1,4‐dioxanyl or 1,2‐oxazinanyl, for example, or a 7‐membered ring, such as azepanyl, 1,4‐diazepanyl or 1,4‐oxazepanyl, for example. Particularly, “4‐ to 6‐membered heterocycloalkyl” means a 4‐ to 6‐membered heterocycloalkyl as defined supra containing one ring nitrogen atom and optionally one further ring heteroatom from the series: N, O, S. More particularly, “5‐ or 6‐membered heterocycloalkyl” means a monocyclic, saturated heterocycle with 5 or 6 ring atoms in total, containing one ring nitrogen atom and optionally one further ring heteroatom from the serie s: N, O. The term “4‐ to 7‐memebered azacycloalkyl” mea ns a monocyclic saturated heterocycly with 4, 5, 6 or 7 ring atoms in total which is attached to the rest of the molecule via the nitrogen atom and whi ch optionally contains one more heteroatom selected from nitrogen and oxygen. Said 4‐ to 7‐membered azacycloalkyl group, without being limited thereto, can be a 4‐membered ring, such as azetidin‐1‐yl, for example; or a 5 ‐membered ring, such as pyrrolidin‐1‐yl, imidazol idin‐1‐ yl, pyrazolidin‐1‐yl, 1,2‐oxazolidin‐2‐yl or 1,3‐oxazolidin‐3‐yl, for example; or a 6‐membered ring, such as piperidin‐1‐yl, morpholin‐4‐yl, piperazin‐1‐yl or 1,2‐oxazinan‐2‐yl, for example, or a 7‐membered ring, such as azepan‐1‐yl, 1,4‐diaze pan‐1‐yl or 1,4‐oxazepan‐4‐yl, for example. The term “5‐ to 10‐membered heterocycloalkenyl” means a monocyclic, unsaturated, non‐aromatic heterocycle with 5, 6, 7, 8, 9 or 10 ring atoms i n total, which contains one or two double bonds and one or two identical or different ring heteroatoms f rom the series: N, O, S; it being possible for sai d heterocycloalkenyl group to be attached to the rest of the molecule via any one of the carbon atoms or, if present, a nitrogen atom. Said heterocycloalkenyl group is, for example, 4H‐pyranyl, 2H‐pyranyl, 2,5‐dihydro‐1H‐pyrrolyl, [1,3]dioxolyl, 4H‐[1,3,4]thiadiazinyl, 2,5‐dihydrofuranyl, 2,3‐dihydrofuranyl, 2,5‐dihydrothiophenyl, 2,3‐dihydrothiophenyl, 4,5‐dihydrooxazolyl or 4H‐[1 ,4]thiazinyl. The term “heterospirocycloalkyl” means a bicyclic, saturated heterocycle with 6, 7, 8, 9, 10 or 11 r ing atoms in total, in which the two rings share one common ring carbon atom, which “heterospirocycloalkyl” contains one, two or three identical or different ring heteroatoms from the series: N, O, S; it being possible for said heteros pirocycloalkyl group to be attached to the rest of the molecule via any one of the carbon atoms, except th e spiro carbon atom, or, if present, a nitrogen atom. Said heterospirocycloalkyl group is, for example, azaspiro[2.3]hexyl, azaspiro[3.3]heptyl, oxaazaspiro[3.3]heptyl, thiaazaspiro[3.3]heptyl, oxaspiro[3.3]heptyl, oxazaspiro[5.3]nonyl, oxazaspiro[4.3]octyl, azaspiro[4,5]decyl, oxazaspiro [5.5]undecyl, diazaspiro[3.3]heptyl, thiazaspiro[3.3]heptyl, thiazaspiro[4.3]octyl, azaspiro[5. 5]undecyl, or one of the further homologous scaffolds such as spiro[3.4]‐, spiro[4.4]‐, spiro[2.4]‐, spiro[2.5]‐, spiro[2.6]‐, spiro[3.5]‐, spiro[3.6]‐, spiro[4.5]‐ and spiro[4.6]‐. The term “6‐ to 10‐membered azaspirocycloalkyl” means a bicyclic, saturated heterocycle with 6, 7, 8, 9 or 10 ring atoms in total, in which the two rings share one common ring carbon atom and which is bound to the rest of the molecule via the nitro gen atom and which azaspirocycloalkyl may contain up to 2 further heteroatoms selected from nitrogen a nd oxygen. Said azaspirocycloalkyl is for example, azaspiro[2.3]hexyl, azaspiro[3.3]heptyl, oxaazaspiro[3.3]heptyl, oxazaspiro[5.3]nonyl, oxazaspiro[4.3]octyl, azaspiro[4,5]decyl, oxazaspiro[5.5]undecyl, diazaspiro[3.3]heptyl, triazaspiro [3.4]octyl or one of the further homologous scaffolds such as spiro[3.4]‐, spiro[4.4]‐, spiro[2 .4]‐, spiro[2.5]‐, spiro[2.6]‐, spiro[3.5]‐, spi ro[3.6]‐ and spiro[4.5]‐, whereby these azaspirocycloalkyl groups are always bound via the nitrogen atom to the rest of the molecule. Of these groups preference is given to 2‐oxa‐6‐ azaspiro[3.3]hept‐6‐yl and 2,5,7‐triazaspiro[3.4]oct an‐ 2‐yl. The term “fused heterocycloalkyl” means a bicyclic , saturated heterocycle with 6, 7, 8, 9 or 10 ring atoms in total, in which the two rings share two a djacent ring atoms, which “fused heterocycloalkyl” contains one or two identical or different ring hete roatoms from the series: N, O, S; it being possible for said fused heterocycloalkyl group to be attached to the rest of the molecule via any one of the carbon atoms or, if present, a nitrogen atom. Said fused heterocycloalkyl group is, for example, azabicyclo[3.3.0]octyl, azabicyclo[4.3.0]nonyl, diazabicyclo[4.3.0]nonyl, oxazabicyclo[4.3.0]nonyl, thiazabicyclo[4.3.0]nonyl or azabicyclo[4.4.0]decyl. The term “bridged heterocycloalkyl” means a bicycl ic, saturated heterocycle with 7, 8, 9 or 10 ring atoms in total, in which the two rings share two c ommon ring atoms which are not adjacent, which “bridged heterocycloalkyl” contains one or two identical or different ring heteroatoms from the series: N, O, S; it being possible for said bridged heterocycloalkyl group to be attached to the rest of the molecule via any one of the carbon atoms, except the spiro carbon atom, or, if present, a nitrogen atom. Said bridged heterocycloalkyl group is, for example, azabicyclo[2.2.1]heptyl, oxazabicyclo[2.2.1]heptyl, thiazabicyclo[2.2.1]heptyl, diazabicyclo[2.2.1]heptyl, azabicyclo‐ [2.2.2]octyl, diazabicyclo[2.2.2]octyl, oxazabicyclo[2.2.2]octyl, thiazabicyclo[2.2.2]octyl, azabi‐ cyclo[3.2.1]octyl, diazabicyclo[3.2.1]octyl, oxazabicyclo[3.2.1]octyl, thiazabicyclo[3.2.1]octyl, azabicyclo[3.3.1]nonyl, diazabicyclo[3.3.1]nonyl, oxazabicyclo[3.3.1]nonyl, thiazabicyclo[3.3.1]nonyl, azabicyclo[4.2.1]nonyl, diazabicyclo[4.2.1]nonyl, oxazabicyclo[4.2.1]nonyl, thiazabicyclo[4.2.1]nonyl, azabicyclo[3.3.2]decyl, diazabicyclo[3.3.2]decyl, oxazabic yclo[3.3.2]decyl, thiazabicyclo[3.3.2]decyl or azabicyclo[4.2.2]decyl. The term “heteroaryl” means a monovalent, monocycl ic, bicyclic or tricyclic aromatic ring having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms (a “5‐ to 14‐membered heteroaryl” group), particularly 5, 6, 9 or 10 ring atoms, which contains at least one ring het eroatom and optionally one, two or three further ring heteroatoms from the series: N, O and/or S, and which is bound via a ring carbon atom or optionally via a ring nitrogen atom (if allowed by valency). Said heteroaryl group can be a 5‐membered heteroary l group, such as, for example, thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl or tetrazolyl; or a 6‐membered heteroaryl group, such as, for example, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl; or a tricyclic heteroaryl group, such as, for example, carbazolyl, acridinyl or phenazinyl; a 8‐membered he teroaryl group, such as for example 6,7‐dihydro‐ 5H‐pyrrolo[1,2‐a]imidazolyl or a 9‐membered hetero aryl group, such as, for example, benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl, indolizinyl, thienopyridinyl, 1H‐pyrrolo[2,3‐b]pyridin yl or purinyl; or a 10‐membered heteroaryl group, such as, for example, quinolinyl, quinazolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinoxalinyl or pteridinyl. In general, and unless otherwise mentioned, the heteroaryl or heteroarylene groups include all possible isomeric forms thereof, e.g.: tautomers and positional isomers with respect to the point of linkage to the rest of the molecule. Thus, for some illustrative non‐restricting examples, the term pyridinyl includes pyridin‐2‐yl, pyridin‐3‐yl and pyridin‐4‐yl; or the term thienyl includes thien‐2‐yl and thien‐3‐yl. A C4 to C12 carbocyclic, heterocyclic, optionally bicyclic, optionally aromatic or optionally heteroaromatic ring system, wherein in a bicyclic, aromatic or heteroaromatic ring system one or two double bonds can be hydrogenated is selected from the group of the substituents phenyl, naphthyl, 1,2,3,4‐tetrahydronaphthyl, 1,3‐benzodioxolyl, quinolinyl, isoquinolinyl, 2,3‐dihydro‐1,4‐ benzodioxinyl, imidazo[1,2‐a]pyridinyl, furanyl, thienyl, pyridinyl, 2H‐1,4‐benzoxazinyl‐3(4H)‐one, 2,1,3‐benzothiadiazolyl, 1‐benzofuranyl, 1‐benzothie nyl, 1H‐indazolyl, 1H‐indolyl, 1H‐benzimidazolyl, 1,3‐benzothiazolyl, thieno[2,3‐b]pyridinyl, thieno[2,3‐c]pyridinyl, thieno[3,2‐c]pyridinyl, pyrimidinyl, 1H‐pyrazolyl, 6,7‐dihydro‐5H‐pyrrolo[1,2‐a]imidazolyl, 1,2‐oxazolyl, 1H‐imidazolyl, 1,3,4‐oxadiazolyl, 1H‐tetrazolyl, 1H‐pyrrolyl, 1H‐pyrrolo[2,3‐b]pyrid inyl or 3,4‐dihydro‐2H‐1,4‐benzoxazinyl. Particularly, the heteroaryl group is a quinolinyl, isoquinolinyl, imidazo[1,2‐a]pyridinyl, furanyl, thienyl, pyridinyl, 2,1,3‐benzothiadiazolyl, 1‐benzofuranyl, 1‐benzothiophenyl, 1H‐indazolyl, 1H‐ indolyl, 1H‐benzimidazolyl, 1,3‐benzothiazolyl, thieno[2,3‐b]pyridinyl, thieno[2,3‐c]pyridinyl, thieno[3,2‐c]pyridinyl, pyrimidinyl, 1H‐pyrazolyl, 6,7‐dihydro‐5H‐pyrrolo[1,2‐a]imidazolyl, 1,2‐ oxazolyl, 1H‐imidazolyl, 1,3,4‐oxadiazolyl, 1H‐tetrazolyl, 1H‐pyrrolyl, 1H‐pyrrolo[2,3‐b]pyridinyl or 3,4‐dihydro‐2H‐1,4‐benzoxazinyl group. In composite substituents such as C 1 ‐C 6 ‐haloalkyl, C 1 ‐C 4 ‐haloalkyl, C 1 ‐C 6 ‐haloalkoxy, ‐(CH 2 )‐heteroaryl, heteroaryloxy, ‐O‐(CH 2 ) x ‐heteroaryl, ‐O‐(CH 2 ) z ‐heteroaryl, O‐(CH 2 )‐4‐ to 7‐membered heterocycloalkyl, bicyclic heteroaryl, C 1 ‐C 6 ‐hydroxyalkyl, ‐O‐(CH 2 ) x ‐C 3 ‐C 8 ‐cycloalkyl, O‐(CH 2 ) x ‐phenyl, ‐ O‐(CH 2 ) x ‐heterocyclyl and C 3 ‐C 8 ‐cycloalkyloxy the definition of the residue to which the further substituent is attached is the same as given for th e residues which do not bear a further substituent, e.g. in C 1 ‐C 6 ‐haloalkyl the C 1 ‐C 6 ‐alkyl has the same meanings as given for th e C 1 ‐C 6 ‐alkyl earlier. The term “C 1 ‐C 6 ”, as used in the present text, e.g. in the context of the definition of “C 1 ‐C 6 ‐alkyl”, “C 1 ‐C 6 ‐haloalkyl”, “C 1 ‐C 6 ‐hydroxyalkyl”, “C 1 ‐C 6 ‐alkoxy” or “C 1 ‐C 6 ‐haloalkoxy” means an alkyl group having a finite number of carbon atoms of 1 to 6, i.e. 1, 2, 3, 4, 5 or 6 carbon atoms. Further, as used herein, the term “C 3 ‐C 8 ”, as used in the present text, e.g. in the context of the definition of “C 3 ‐C 8 ‐cycloalkyl”, means a cycloalkyl group having a finite number of carbon atoms of 3 to 8, i.e. 3, 4, 5, 6, 7 or 8 carbon atoms. When a range of values is given, said range encompa sses each value and sub‐range within said range. For example: "C 1 ‐C 6 " encompasses C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 1 ‐C 6 , C 1 ‐C 5 , C 1 ‐C 4 , C 1 ‐C 3 , C 1 ‐C 2 , C 2 ‐C 6 , C 2 ‐C 5 , C 2 ‐C 4 , C 2 ‐C 3 , C 3 ‐C 6 , C 3 ‐C 5 , C 3 ‐C 4 , C 4 ‐C 6 , C 4 ‐C 5 , and C 5 ‐C 6 ; "C 2 ‐C 6 " encompasses C 2 , C 3 , C 4 , C 5 , C 6 , C 2 ‐C 6 , C 2 ‐C 5 , C 2 ‐C 4 , C 2 ‐C 3 , C 3 ‐C 6 , C 3 ‐C 5 , C 3 ‐C 4 , C 4 ‐C 6 , C 4 ‐C 5 , and C 5 ‐C 6 ; "C 3 ‐C 10 " encompasses C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 3 ‐C 10 , C 3 ‐C 9 , C 3 ‐C 8 , C 3 ‐C 7 , C 3 ‐C 6 , C 3 ‐C 5 , C 3 ‐C 4 , C 4 ‐C 10 , C 4 ‐C 9 , C 4 ‐C 8 , C 4 ‐C 7 , C 4 ‐C 6 , C 4 ‐C 5 , C 5 ‐C 10 , C 5 ‐C 9 , C 5 ‐C 8 , C 5 ‐C 7 , C 5 ‐C 6 , C 6 ‐C 10 , C 6 ‐C 9 , C 6 ‐C 8 , C 6 ‐C 7 , C 7 ‐C 10 , C 7 ‐C 9 , C 7 ‐C 8 , C 8 ‐C 10 , C 8 ‐C 9 and C 9 ‐C 10 ; "C 3 ‐C 8 " encompasses C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 3 ‐C 8 , C 3 ‐C 7 , C 3 ‐C 6 , C 3 ‐C 5 , C 3 ‐C 4 , C 4 ‐C 8 , C 4 ‐C 7 , C 4 ‐C 6 , C 4 ‐C 5 , C 5 ‐C 8 , C 5 ‐C 7 , C 5 ‐C 6 , C 6 ‐C 8 , C 6 ‐C 7 and C 7 ‐C 8 ; "C 3 ‐C 6 " encompasses C 3 , C 4 , C 5 , C 6 , C 3 ‐C 6 , C 3 ‐C 5 , C 3 ‐C 4 , C 4 ‐C 6 , C 4 ‐C 5 , and C 5 ‐C 6 ; "C 4 ‐C 8 " encompasses C 4 , C 5 , C 6 , C 7 , C 8 , C 4 ‐C 8 , C 4 ‐C 7 , C 4 ‐C 6 , C 4 ‐C 5 , C 5 ‐C 8 , C 5 ‐C 7 , C 5 ‐C 6 , C 6 ‐C 8 , C 6 ‐C 7 and C 7 ‐C 8 ; "C 4 ‐C 7 " encompasses C 4 , C 5 , C 6 , C 7 , C 4 ‐C 7 , C 4 ‐C 6 , C 4 ‐C 5 , C 5 ‐C 7 , C 5 ‐C 6 and C 6 ‐C 7 ; "C 4 ‐C 6 " encompasses C 4 , C 5 , C 6 , C 4 ‐C 6 , C 4 ‐C 5 and C 5 ‐C 6 ; "C 5 ‐C 10 " encompasses C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 5 ‐C 10 , C 5 ‐C 9 , C 5 ‐C 8 , C 5 ‐C 7 , C 5 ‐C 6 , C 6 ‐C 10 , C 6 ‐C 9 , C 6 ‐C 8 , C 6 ‐C 7 , C 7 ‐C 10 , C 7 ‐C 9 , C 7 ‐C 8 , C 8 ‐C 10 , C 8 ‐C 9 and C 9 ‐C 10 ; "C 6 ‐C 10 " encompasses C 6 , C 7 , C 8 , C 9 , C 10 , C 6 ‐C 10 , C 6 ‐C 9 , C 6 ‐C 8 , C 6 ‐C 7 , C 7 ‐C 10 , C 7 ‐C 9 , C 7 ‐C 8 , C 8 ‐C 10 , C 8 ‐C 9 and C 9 ‐C 10 . As used herein, the term “leaving group” means a n atom or a group of atoms that is displaced in a chemical reaction as stable species taking with it t he bonding electrons. In particular, such a leaving group is selected from the group comprising: halide, in particular fluoride, chloride, bromide or iodide, (methylsulfonyl)oxy, [(trifluoromethyl)sulfonyl]oxy, [(nonafluorobutyl)sulfonyl]oxy, (phenylsulfonyl)oxy, [(4‐methylphenyl)sulfonyl]oxy, [(4‐bromophenyl)sulfonyl]oxy, [(4‐nitrophenyl)sulfonyl]oxy, [(2‐nitrophenyl)sulfonyl]oxy, [(4‐isopropylphenyl)sulfonyl]oxy, [(2,4,6‐triisopropylphenyl)sulfonyl]oxy, [(2,4,6‐trimethylphenyl)sulfonyl]oxy, [(4‐tert‐butyl‐ phenyl)sulfonyl]oxy and [(4‐methoxyphenyl)sulfonyl]oxy. It is possible for the compounds of general formula (I) to exist as isotopic variants. The invention therefore includes one or more isotopic variant(s) of the compounds of general formula (I), particularly deuterium‐containing compounds of general formula (I). The term “Isotopic variant” of a compound or a reagent is defined as a compound exhibiting an unnatural proportion of one or more of the isotopes that constitute such a compound. The term “Isotopic variant of the compound of general formula (I)” is defined as a compound of general formula (I) exhibiting an unnatural proportion of one or more of the isotopes that constitute such a compound. The expression “unnatural proportion” means a prop ortion of such isotope which is higher than its natural abundance. The natural abundances of isotopes to be applied in this context are described in “Isotopic Compositions of the Elements 1997”, Pure Appl. Chem., 70(1), 217‐235, 1998. Examples of such isotopes include stable and radioactive isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine and iodine, such as 2 H (deuterium), 3 H (tritium), 11 C, 13 C, 14 C, 15 N, 17 O, 18 O, 32 P, 33 P, 33 S, 34 S, 35 S, 36 S, 18 F, 36 Cl, 82 Br, 123 I, 124 I, 125 I, 129 I and 131 I, respectively. With respect to the treatment and/or prophylaxis of the disorders specified herein the isotopic variant(s) of the compounds of general formula (I) preferably contain deuterium (“deuterium‐ containing compounds of general formula (I)”). Isotopic variants of the compounds of general formula (I) in which one or more radioactive isotope s, such as 3 H or 14 C, are incorporated are useful e.g. in drug and/or substrate tissue distribution stu dies. These isotopes are particularly preferred for the ease of their incorporation and detectability. Po sitron emitting isotopes such as 18 F or 11 C may be incorporated into a compound of general formula (I). These isotopic variants of the compounds of general formula (I) are useful for in vivo imaging applications. Deuterium‐containing and 13 C‐ containing compounds of general formula (I) can be used in mass spectrometry analyses in the context of preclinical or clinical studies. Isotopic variants of the compounds of general formula (I) can generally be prepared by methods known to a person skilled in the art, such as thos e described in the schemes and/or examples herein, by substituting a reagent for an isotopic variant of said reagent, preferably for a deuterium‐ containing reagent. Depending on the desired sites of deuteration, in some cases deuterium from D 2 O can be incorporated either directly into the compounds or into reagents that are useful for synthesizing such compounds. Deuterium gas is also a useful reagent for incorporating deuterium into molecules. Catalytic deuteration of olefinic bonds and acetylenic bonds is a rapid route for incorporation of deuterium. Metal catalysts (i.e. Pd, Pt, and Rh) in the presence of deuterium gas can be used to directly exchange deuterium for hydrogen in functional groups containing hydrocarbons. A variety of deuterated reagents and synthetic building blocks are commercially available from companies such as for example C/D/N Isotopes, Quebec, Canada; Cambridge Isotope Laboratories Inc., Andover, MA, USA; and CombiPhos Catalysts, Inc. , Princeton, NJ, USA. The term “deuterium‐containing compound of general formula (I)” is defined as a compound of general formula (I), in which one or more hydrogen atom(s) is/are replaced by one or more deuterium atom(s) and in which the abundance of deuterium at each deuterated position of the compound of general formula (I) is higher than the natural abundance of deuterium, which is about 0.015%. Particularly, in a deuterium‐containing compound of general formula (I) the abundance of deuterium at each deuterated position of the compound of general formula (I) is higher than 10%, 20%, 30%, 40%, 50%, 60%, 70% or 80%, preferably hig her than 90%, 95%, 96% or 97%, even more preferably higher than 98% or 99% at said position(s). It is understood that the abundance of deuterium at each deuterated position is independent of the abundance of deuterium at other deuterated position(s). The selective incorporation of one or more deuterium atom(s) into a compound of general formula (I) may alter the physicochemical properties (such as for example acidity [C. L. Perrin, et al., J. Am. Chem. Soc., 2007, 129, 4490], basicity [C. L. Perrin et al., J. Am. Chem. Soc., 2005, 127, 9641 ], lipophilicity [B. Testa et al., Int. J. Pharm., 1984, 19(3), 271]) and/or the metabolic profile of the molecule and may result in changes in the ratio of parent compound to metabolites or in the amounts of metabolites formed. Such changes may result in certain therapeutic advantages and hence may be preferred in some circumstances. Reduced rates of metabolism and metabolic switching, where the ratio of metabolites is changed, have been reported (A. E. Mutlib et al., Toxicol. Appl. Pharmacol., 2000, 169, 102). These changes in the exposure to parent drug and metabolites can have important consequences with respect to the pharmacodynamics, tolerability and efficacy of a deuterium‐containing compound of general formula (I). In some cases deuterium substitution reduces or eliminates the formation of an undesired or toxic metabolite and enhances the formation of a desired metabolite (e.g. Nevirapine: A. M. Sharma et al., Chem. Res. Toxicol., 2013, 26, 410; Efavirenz: A. E. Mutlib et al., Toxicol. Appl. Pharm acol., 2000, 169, 102). In other cases the major effect of deuteration is to reduce the rate of syst emic clearance. As a result, the biological half‐li fe of the compound is increased. The potential clinical benefits would include the ability to maintain similar systemic exposure with decreased peak levels and increased trough levels. This could result in lower side effects and enhanced efficacy, depending o n the particular compound’s pharmacokinetic/ pharmacodynamic relationship. ML‐337 (C. J. Wenthur et al., J. Med. Chem., 2013, 56, 5208) and Odanacatib (K. Kassahun et al., WO2012/112363) are ex amples for this deuterium effect. Still other cases have been reported in which reduced rates of metabolism result in an increase in exposure of the drug without changing the rate of systemic clear ance (e.g. Rofecoxib: F. Schneider et al., Arzneim. Forsch. / Drug. Res., 2006, 56, 295; Telaprevir: F. Maltais et al., J. Med. Chem., 2009, 52, 7993). Deuterated drugs showing this effect may have reduced dosing requirements (e.g. lower number of doses or lower dosage to achieve the desired effect) and/or may produce lower metabolite loads. A compound of general formula (I) may have multiple potential sites of attack for metabolism. To optimize the above‐described effects on physicochemical properties and metabolic profile, deuterium‐containing compounds of general formula (I) having a certain pattern of one or more deuterium‐hydrogen exchange(s) can be selected. Parti cularly, the deuterium atom(s) of deuterium‐ containing compound(s) of general formula (I) is/are attached to a carbon atom and/or is/are located at those positions of the compound of general formul a (I), which are sites of attack for metabolizing enzymes such as e.g. cytochrome P 450 . In another embodiment the present invention concerns a deuterium‐containing compound of general formula (I), in which one, two or three of the hydrogen atom(s) in either one or both of the methyl groups shown in general formula (I) is/are re placed with a deuterium atom. Also the hydrogen atom on the carbon atom between t he nitrogen atom and the group A1 can be replaced with a deuterium atom either as the single replacement of a hydrogen by a deuterium or in addition to the beforementioned replacements in either one or both of the methyl groups shown in general formula (I). Where the plural form of the word compounds, salts, polymorphs, hydrates, solvates and the like, is used herein, this is taken to mean also a single c ompound, salt, polymorph, isomer, hydrate, solvate or the like. By "stable compound' or "stable structure" is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent. The compounds of the present invention contain at least one or optionally even more asymmetric centres, depending upon the location and nature of the various substituents desired. It is possible that one or more asymmetric carbon atoms are present in the (R) or (S) configuration, which can result in racemic mixtures in the case of a single asymmetric centre, and in diastereomeric mixtures in the case of multiple asymmetric centres. In certa in instances, it is possible that asymmetry also be present due to restricted rotation about a given bon d, for example, the central bond adjoining two substituted aromatic rings of the specified compounds. Preferred isomers are those which produce the more d esirable biological activity. Separated, pure or partially purified isomers and stereoisomers or racemic or diastereomeric mixtures of the compounds of the present invention are also included within the scope of the present invention. The purification and the separation of such materials can be accomplished by standard techniques known in the art. The optical isomers can be obtained by resolution of the racemic mixtures according to conventional processes, for example, by the formation of diastereo isomeric salts using an optically active acid or base or formation of covalent diastereomers. Examples of appropriate acids are tartaric, diacetyltartaric, ditoluoyltartaric and camphorsulfonic acid. Mixtures of diastereoisomers can be separated into their individual diastereomers on the basis of their physical and/or chemical differences by methods known in the art, for example, by chromatography or fractional crystallisation. The optically active bases or acids are then liberated from the separated diastereomeric salts. A different process for separati on of optical isomers involves the use of chiral chromatography (e.g., HPLC columns using a chiral phase), with or without conventional derivatisation, optimally chosen to maximise the separation of the enantiomers. Suitable HPLC columns using a chiral phase are commercially availab le, such as those manufactured by Daicel, e.g., Chiracel OD and Chiracel OJ, for example, among many others, which are all routinely selectable. Enzymatic separations, with or without derivatisation, are also useful. The optically active compounds of the present invention can likewise be obtained by chiral syntheses utilizing op tically active starting materials. In order to distinguish different types of isomers f rom each other reference is made to IUPAC Rules Section E (Pure Appl Chem 45, 11‐30, 1976). The present invention includes all possible stereoisom ers of the compounds of the present invention as single stereoisomers, or as any mixture of said stereoisomers, e.g. (R)‐ or (S)‐ isomers, in any ratio. Isolation of a single stereoisomer, e.g. a single en antiomer or a single diastereomer, of a compound of the present invention is achieved by any suitable state of the art method, such as chromatography, especially chiral chromatography, for e xample. Further, it is possible for the compounds of the pr esent invention to exist as tautomers. For example, any compound of the present invention which contains an imidazopyridine moiety as a heteroaryl group for example can exist as a 1H tautomer, or a 3H tautomer, or even a mixture in any amount of the two tautomers, namely: The present invention includes all possible tautomers of the compounds of the present invention as single tautomers, or as any mixture of said tautomer s, in any ratio. Further, the compounds of the present invention can exist as N‐oxides, which are defined in that at least one nitrogen of the compounds of the present invention is oxidised. The present invention includes all such possible N‐oxides. The present invention also covers useful forms of the compounds of the present invention, such as metabolites, hydrates, solvates, prodrugs, salts, in particular pharmaceutically acceptable salts, and/or co‐precipitates. The compounds of the present invention can exist as a hydrate, or as a solvate, wherein the compounds of the present invention contain polar solv ents, in particular water, methanol or ethanol for example, as structural element of the crystal lattice of the compounds. It is possible for the amount of polar solvents, in particular water, to ex ist in a stoichiometric or non‐stoichiometric ratio. In the case of stoichiometric solvates, e.g. a hydra te, hemi‐, (semi‐), mono‐, sesqui‐, di‐, tri ‐, tetra‐, penta‐ etc. solvates or hydrates, respectively, are possible. The present invention includes all such hydrates or solvates. Further, it is possible for the compounds of the pr esent invention to exist in free form, e.g. as a f ree base, or as a free acid, or as a zwitterion, or t o exist in the form of a salt. Said salt may be any salt, either an organic or inorganic addition salt, particularly any pharmaceutica lly acceptable organic or inorganic addition salt, which is customarily used in pharmacy, or which is used, for example, for isolating or purifying the compounds of the present invention. The term “pharmaceutically acceptable salt" refers t o an inorganic or organic acid addition salt of a compound of the present invention. For example, see S. M. Berge, et al. “ Pharmaceutical Salts,” J. Pharm. Sci. 1977, 66, 1‐19. A suitable pharmaceutically acceptable salt of the co mpounds of the present invention may be, for example, an acid‐addition salt of a compound of th e present invention bearing a nitrogen atom, in a chain or in a ring, for example, which is sufficiently basic, such as an acid‐addition salt with an inorganic acid, or “mineral acid”, such as hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic, bisulfuric, phosphoric, or nitric acid, for example, or with an organic acid, such as formic, acetic, acetoacetic, pyruvic, trifluoroacetic, propionic, butyric, hexanoic, heptanoic, undecanoic, lauric, benzoic, salicylic, 2‐(4‐hydroxybenzoyl)‐benzoic, camphoric, cinnamic, cyclopentanepropionic, digluconic, 3‐hydroxy‐2‐naphthoic, nicotinic, pamoic, pectinic, 3‐phenylpropionic, pivalic, 2‐ hydroxyethanesulfonic, itaconic, trifluoromethanesulfonic, dodecylsulfuric, ethanesulfonic, benzenesulfonic, para‐toluenesulfonic, methanesulfonic, 2‐naphthalenesulfonic, naphthalinedisulfonic, camphorsulfonic acid, citric, tartaric, stearic, lactic, oxalic, malonic, succinic, malic, adipic, alginic, maleic, fumaric, D‐gluconic, mandelic, ascorbic, glucoheptanoic, glycerophosphoric, aspartic, sulfosalicylic, or thiocyan ic acid, for example. Further, another suitably pharmaceutically acceptable s alt of a compound of the present invention which is sufficiently acidic, is an alkali metal sal t, for example a sodium or potassium salt, an alkal ine earth metal salt, for example a calcium, magnesium o r strontium salt, or an aluminium or a zinc salt, or an ammonium salt derived from ammonia or from an organic primary, secondary or tertiary amine having 1 to 20 carbon atoms, such as ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, diethylaminoethanol, tris(hydroxymethyl)aminomethane, procaine, dibenzylamine, N‐methylmorpholine, arginine, lysine, 1,2‐ethylenediamine, N‐methylpiperidine, N‐ methyl‐glucamine, N,N‐dimethyl‐glucamine, N‐ethyl‐glucamine, 1,6‐hexanediamine, glucosamine, sarcosine, serinol, 2‐amino‐1,3‐propanediol, 3‐am ino‐1,2‐propanediol, 4‐amino‐1,2,3‐butanetriol, or a salt with a quarternary ammonium ion having 1 to 20 carbon atoms, such as tetramethylammonium, tetraethylammonium, tetra(n‐propyl)ammonium, tetra(n‐butyl)ammonium, N‐benzyl‐N,N,N‐trimethylammonium, choline or benzal konium. Those skilled in the art will further recognise that it is possible for acid addition salts of the cla imed compounds to be prepared by reaction of the compound s with the appropriate inorganic or organic acid via any of a number of known methods. Alternat ively, alkali and alkaline earth metal salts of acidic compounds of the present invention are prepared by reacting the compounds of the present invention with the appropriate base via a variety of known methods. The present invention includes all possible salts of the compounds of the present invention as single salts, or as any mixture of said salts, in any rat io. In the present text, in particular in the Experiment al Section, for the synthesis of intermediates and of examples of the present invention, when a compound is mentioned as a salt form with the corresponding base or acid, the exact stoichiometric composition of said salt form, as obtained by the respective preparation and/or purification process, is, in most cases, unknown. Unless specified otherwise, suffixes to chemical names or structural formulae relating to salts, such as "hydrochloride", "trifluoroacetate", "sodium salt", or "x HCl", "x CF 3 COOH", "x Na + ", for example, mean a salt form, the stoichiometry of which salt f orm not being specified. This applies analogously to cases in which synthesis intermediates or example compounds or salts thereof have been obtained, by the preparation and/or purification processes described, as solvates, such as hydrates, with (if defined) unknown stoichiom etric composition. As used herein, the term “in vivo hydrolysable ester” means an in vivo hydrolysable ester of a compound of the present invention containing a carboxy or hydroxy group, for example, a pharmaceutically acceptable ester which is hydrolysed in the human or animal body to produce the parent acid or alcohol. Suitable pharmaceutically acce ptable esters for carboxy include for example alkyl, cycloalkyl and optionally substituted phenylalky l, in particular benzyl esters, C 1 ‐C 6 alkoxymethyl esters, e.g. methoxymethyl, C 1 ‐C 6 alkanoyloxymethyl esters, e.g. pivaloyloxymethyl, phthalidyl esters, C 3 ‐C 8 cycloalkoxy‐carbonyloxy‐C 1 ‐C 6 alkyl esters, e.g. 1‐cyclohexylcarbonyloxyethyl ; 1,3‐dioxolen‐2‐ onylmethyl esters, e.g. 5‐methyl‐1,3‐dioxolen‐2‐onylmethyl ; and C 1 ‐C 6 ‐alkoxycarbonyloxyethyl esters, e.g. 1‐methoxycarbonyloxyethyl, it being poss ible for said esters to be formed at any carboxy group in the compounds of the present invention. An in vivo hydrolysable ester of a compound of the present invention containing a hydroxy group includes inorganic esters such as phosphate esters and [alpha]‐acyloxyalkyl ethers and related compounds which as a result of the in vivo hydrolysis of the ester breakdown to give the parent hydroxy group. Examples of [alpha]‐acyloxyalkyl ethers include acetoxymethoxy and 2,2‐ dimethylpropionyloxymethoxy. A selection of in vivo hydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl and N‐(dialkylaminoethyl)‐N‐alkylcarbamoyl (to give carbamates), dialkylaminoacetyl and carboxyacetyl. The present invention covers all such esters. Furthermore, the present invention includes all possible crystalline forms, or polymorphs, of the compounds of the present invention, either as single polymorph, or as a mixtu re of more than one polymorph, in any ratio. Moreover, the present invention also includes prodrugs of the compounds according to the invention. The term “prodrugs” here designates compounds which themselves can be biologically active or inactive, but are converted (for example metabolically or hydrolytically) into compounds according to the invention during their residence tim e in the body. In accordance with other embodiments, the present inv ention covers the following compounds. A) A compound of formula I or Ia, wherein R 1 is selected from ‐H, ‐Br, ‐OH, ‐NO 2 , ‐CH 3 , , ‐O‐CH 3 , ‐O‐CH 2 ‐CH 3 , ‐O‐CH(CH 3 ) 2 , ‐O‐(CH 2 ) 3 CH 3 , ‐O‐(CH 2 ) 2 CH(CH 3 ) 2 , ‐O‐CH 2 ‐phenyl, ‐O‐(CH 2 ) 2 ‐O‐CH 3 , ‐O‐(CH 2 ) 2 ‐S(O) 2 ‐CH 3 , ‐CH 2 ‐OH, ‐C(CH 3 ) 2 ‐OH, ‐C(O)OH, ‐C(O)OCH 3 , ‐NH 2 , ‐NH(CH 3 ), ‐N(CH 3 ) 2 , ‐NH‐(CH 2 ) 2 ‐NH‐C(O)‐CH 3 , ‐NH‐(CH 2 ) 2 ‐morpholino, ‐NH‐C(O)‐CH 3 , ‐NH‐C(O)‐NH‐CH 3 , ‐NH‐C(O)‐N(CH 3 ) 2 , ‐NH‐S(O) 2 ‐CH 3 , ‐N=S(O)(CH 3 ) 2 , y is 1 or 2; A is selected from the group consisting of C 6‐10 aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; R 2 is each independently selected from the group consisting of C 1‐4 alkyl, C 2‐4 alkenyl, C 2‐ 4 alkinyl, C 1‐4 haloalkyl, hydroxy‐C 1‐4 alkyl, hydroxy‐C 1‐4 haloalkyl, C 3‐6 cycloalkyl, 3‐6 membered heterocyclyl, hydroxy‐C 3‐6 cycloalkyl, C 1‐4 haloalkyl substituted with a 3‐6 membered heterocyclyl, 3‐6 membered heterocyclyl substituted with hydroxy, halogen, ‐NH 2 , ‐SO 2 ‐C 1‐4 alkyl and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring x is 1, 2 or 3; or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. B) A compound as defined in A (above), wherein A is selected from the group consisting of C 6‐10 ‐aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; R2 is each independently selected from the group consist ing of C 1‐4 ‐alkyl, C 2‐4 ‐alkenyl, C 2‐4 ‐ alkinyl, C 1‐4 ‐haloalkyl, hydroxy‐C 1‐4 ‐alkyl, hydroxy‐C 1‐4 ‐haloalkyl, C 3‐6 ‐cycloalkyl, 3‐6 membered heterocyclyl, hydroxy‐C 3‐6 ‐cycloalkyl, C 1‐4 ‐haloalkyl substituted with a 3‐6 membered heterocyclyl, 3‐6 membered heterocyclyl substituted with hydroxy, halogen, ‐NH 2 , ‐SO 2 ‐C 1‐4 ‐alkyl and the bivalent substituent =O, while =O may only be a substituent in a non‐aromatic ring x is 1, 2 or 3 or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. C) A compound as defined in A or B (above), wherein A is selected from the group consisting of C 6‐10 aryl, 5‐10 membered heteroaryl and 9‐10 membered bicyclic heterocyclyl; x is 1 or 2 R 2 is each independently selected from the group consist ing of C 1‐4 ‐alkyl, C 2‐4 ‐alkinyl, C 1‐4 ‐ haloalkyl, hydroxy‐C 1‐4 ‐haloalkyl, C 1‐4 ‐haloalkyl substituted with a 3‐6 membered heterocyclyl, halogen, and the bivalent substituent =O , while =O may only be a substituent in a non‐aromatic ring or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. D) A compound as defined in A, B or C (above), wherei n and wherein R 3 is selected from the group consisting of C 1‐4 ‐alkyl, C 1‐4 ‐haloalkyl, hydroxy‐C 1‐4 ‐alkyl, hydroxy‐C 1‐4 ‐haloalkyl, C 1‐4 ‐haloalkyl substituted with a 3‐6 membered h eterocyclyl, C 3‐6 ‐cycloalkyl, hydroxy‐C 3‐6 ‐cycloalkyl, 3‐6 membered heterocyclyl, 3‐6 membered hydroxy‐heterocyclyl, halogen and ‐SO 2 ‐C 1‐4 ‐alkyl; R 4 is selected from the group consisting of hydrogen an d ‐NH 2 , R 5 is selected from the group consisting of hydrogen, C 1‐4 ‐alkyl and halogen; or R 3 and R 5 together with the carbon atoms they are attac hed form a 5‐6 membered nonaromatic carbocycle, a 5‐6 membered non‐aromatic heterocycle or a 5‐6 membered heteroaryl, wherein the 5‐6 membered non‐ aromatic carbocycle, 5‐6 membered nonaromatic heterocycle and 5‐6 membered he teroaryl are all optionally substituted by one or more halogen or by an oxo gr oup or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. E) A compound as defined in A, B, C or D (above), wh erein R 3 is selected from the group consisting of C 1‐4 ‐haloalkyl, hydroxy‐C 1‐4 ‐haloalkyl and C 1‐4 ‐ haloalkyl substituted with a 3‐6 membered heterocycl yl; R 4 is hydrogen; R 5 is selected from the group consisting of hydrogen, C 1‐4 ‐alkyl and fluorine; or R 3 and R 5 together with the carbon atoms they are attac hed form a 5‐6 membered nonaromatic carbocycle, a 5‐6 membered non‐aromatic heterocycle or a 5‐6 membered heteroaryl, wherein the 5‐6 membered non‐ aromatic carbocycle, 5‐6 membered nonaromatic heterocycle and 5‐6 membered he teroaryl are all optionally substituted by one or more fluorine or by an oxo g roup or a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. F) A compound as defined in A, B, C, D or E (above), wherein is selected from or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. G) The compound as defined in A, B, C, D, E or F (a bove), wherein V is nitrogen and T is carbon or a stereoisomer, a tautomer, an N‐oxide, a hydrate, a solvate, or a salt thereof, or a mixture of same. H) The compound as defined in A, B, C, D, E, F or G (above), wherein y = 1 and R 1 is selected from or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. I) The compound as defined in A, B, C, D, E, F, G o r H (above), wherein V is nitrogen, T is carbon, y = 1, R 1 is selected from and is selected from or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. J) The compound as defined in A, B, C, D, E, F, G, H or I (above), which is selected from the group consisting of: N‐[(3R)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluo romethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluo romethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) ‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3, 4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) ‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3, 4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐fluoro‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. K) A SOS1 inhibitor compound as described herein or as defined in A, B, C, D, E, F, G, H, I or J (above) for use in the treatment and/or prevention o f cancer, wherein said SOS1 inhibitor compound is administered in combination with at least one other pharmacologically active substance and wherein each of said other pharmacologi cally active substance(s) is selected from the group consisting of: an inhibitor of HRas, NRas or KRAS and mutants thereof, in particular an inhibitor of KRAS‐G12C; an inhibitor of MAP kinases, in particular MEK1, MEK2, ERK1, ERK2, ERK5 and/or of an inhibitor of PI3‐kin ases and mutants thereof; an inhibitor of Tropomyosin Receptor kinases and/or of mutants thereof ; an inhibitor of SHP2 and mutants thereof; inhibitor of EGFR and/or of mutants thereof; an inhibitor of FGFR1 and/or FGFR2 and/or FGFR3 and/or of mutants thereof; an inhibitor of ALK and/or of mutants thereof; an inhibitor of c‐MET and/or of mutants thereof; an i nhibitor of BCR‐ABL and/or of mutants thereof; an inhibitor of ErbB2 (Her2) and/or of muta nts thereof; an inhibitor of AXL and/or of mutants thereof; an inhibitor of A‐Raf and/or B‐R af and/or C‐Raf and/or of mutants thereof; an inhibitor of mTOR and mutants thereof; an inhibitor of IGF1/2 and/or of IGF1‐R; an inhibitor of farnesyl transferase. In accordance with further embodiments, the present i nvention covers the following compounds. A compound of formula I or Ia as defined in A, B, C, D, E, F, G, H or I (above), wherein R 1 is selected from
R 1 can also be selected from
A compound of formula I or Ia as defined in A, B, C, D, E, F, G, H or I (above), wherein R 1 is selected from A compound of formula I as defined in A, B, C, D, E, F, G, H or I (above), wherein A compound of formula I or Ia as defined in A, B, C, D, E, F, G, H or I (above), wherein A compound of formula I or Ia as defined in A, B, C, D, E, F, G, H or I (above), wherein A compound of formula I or Ia as defined in A, B, C, D, E, F, G, H or I (above), wherein A compound of formula I or Ia as defined in A, B, C, D, E, F, G, H or I (above), wherein A compound of formula I or Ia as defined in A, B, C, D, E, F, G, H or I (above), wherein In a particular further embodiment of the first aspe ct, the present invention covers combinations of two or more of the above mentioned embodiments under the heading “further embodiments of the first aspect of the present invention”. Further embodiments of this invention can be presented by the following alternative claim set possibility: 1. A compound of general formula (1) wherein R 1a is selected from the group consisting of 5‐6 membered heteroaryl, 9‐10 membered bicyclic he teroaryl or phenyl, all optionally one or more times substituted by ‐H, ‐OH, ‐CN, ‐NO 2 , ‐NH 2 , halogen, ‐COOH, ‐COO‐CH 3 , ‐SF 5 , (1E)‐2‐ethoxyethenyl, [(tert‐butoxy)carbonyl]amino, 1H‐pyrazol‐1‐yl, 2 (methylamino)ethoxy, oxolan‐3‐ yloxy, (1‐methylpyrrolidin‐3‐yl)oxy, C 1‐6 ‐alkyl optionally substituted one or more times with ‐F and/or ‐OH and/or ‐O‐C 1‐6 ‐alkyl or ‐S‐C 1‐6 ‐alkyl, both optionally substituted one or more times with ‐F; R 2a is selected from the group consisting of −F, −Cl , −OCH 3 , −COOCH 3 , −S(=O) 2 ‐CH 3 , −O‐CH 2 ‐ CH 2 R 9 , −C(=O)‐NHR 3a , 2,5‐dihydrofuran‐3‐yl, 4,5‐dihydrofuran‐ 2‐yl, oxolan‐3‐yl, oxetan‐ 3‐yloxy, cyclopentylamino, 5,6‐dihydro‐2H‐pyran‐ 3‐yl, oxan‐3‐yl, 3,6‐dihydro‐2H‐pyran‐ 4‐yl, 1‐methyl‐1H‐pyrazol‐4‐yl, oxan‐4‐yl, [(oxetan‐2‐yl)methyl]amino, −N(R 3a )‐CH(R 3a )‐ CH 2 ‐R 12 , 1‐methylpiperidin‐4‐yl, 1‐methyl‐1,2,3,6 ‐tetrahydropyridin‐4‐yl, 1‐oxa‐6‐ azaspiro[3.3]heptan‐6‐yl, [(oxolan‐2‐yl)methyl]amin o, 2‐oxa‐6‐azaspiro[3.3]heptan‐6‐yl, (1‐methylpyrrolidin‐3‐yl)oxy, (5‐oxopyrrolidin‐3 yl)amino, 3‐(difluoromethyl)azetidin‐1‐ yl, 2‐oxa‐6‐azaspiro[3.4]octan‐6‐yl, 3‐oxo‐1 ,4‐diazepan‐1‐yl, −R 22 ‐COOC(CH 3 ) 3 , 4‐cyano‐4‐ methylpiperidin‐1‐yl, 2‐oxa‐6‐azaspiro[3.5]nonan 6‐yl, 2‐oxa‐7‐azaspiro[3.5]nonan‐7‐yl, 5‐oxo‐2,6‐diazaspiro[3.4]octan‐2‐yl, 7‐oxo‐2, 6‐diazaspiro[3.4]octan‐2‐yl, 8‐oxo‐2,7‐ diazaspiro[4.4]nonan‐2‐yl, 4‐methyl‐2,3‐dioxo‐1 ,4‐diazepan‐1‐yl, ; R 3a is selected from the group consisting of −H and CH 3 ; R 4a is selected from the group consisting of −H and F; R 5a is selected from the group consisting of −H, −F, −Cl, −Br, −CN, −NO 2 , −OH, −CH 2 OH, −COOH, ‐COO‐CH 3 , −CH 3 , −CF 3 , −CHF 2 , −CF 2 −CH 3 , −CF 2 −CH 2 OH, −CF 2 −C(CH 3 ) 2 OH, −O‐CH 3 , −O‐CH 2 ‐CHF 2 , −O‐CF 3 , ‐O‐CHF 2 , −S‐CF 3 , −SF 5 , (1E)‐2‐ethoxyethenyl, and [(tert‐ butoxy)carbonyl]amino; R 6a is selected from the group consisting of −H, −F, −Cl, −CH 3 , −CHF 2 , −O‐CH 3 , −O‐CHF 2 , 1H‐ pyrazol‐1‐yl, 2‐(methylamino)ethoxy, oxolan‐3‐yl oxy, and (1‐methylpyrrolidin‐3‐yl)oxy; R 7 is selected from the group consisting of −H, −NH 2 , −F, and −Br; R 8 is selected from the group consisting of −H, −CH 3 , and −F; R 9 is selected from the group consisting of −H, −CH 2 ‐CH 3 , and –NH‐CH 3 ; R 10 is selected from the group consisting of R 11 is selected from the group consisting of ‐CH 2 ‐CH 2 ‐CH 2 ‐, ‐CH 2 ‐O‐CH 2 ‐, ‐CH 2 ‐CH 2 ‐O‐, −N(CH 3 )‐CH 2 ‐CH 2 −, ‐CH 2 ‐NH‐CH 2 ‐ and ‐CH 2 ‐N(R 31 )‐CH 2 ‐; R 12 is selected from the group consisting of ‐H, ‐OC H 3 , and ‐N(CH 3 ) 2 ; R 13 is selected from the group consisting of R 14 is selected from the group consisting of −CH 2 ‐C(R 4a ) 2 ‐CH 2 ‐, ‐CH 2 ‐CH 2 ‐C(=O)‐ and ‐CH 2 ‐O‐ C(=O)‐; R 15 is selected from the group consisting of −H, −OH , −F, −OCH 3 , −N(CH 3 ) 2 , −C(=O)‐NH 2 , ‐ COOH, pyrrolidin‐1‐yl, ‐NH‐SO 2 ‐R 34 , ‐N(R 3a )‐CO‐R 35 , and morpholin‐4‐yl; R 16 is selected from the group consisting of −H, −CH 3 , −F, and −CH 2 ‐CH 2 OH; R 17 is selected from the group consisting of −H and N(CH 3 ) 2 ; R 18 is selected from the group consisting of −H and CH=CH 2 ; R 19 is selected from the group consisting of =CH 2 and =O; R 20 is selected from the group consisting of −H and CN; R 21 is selected from the group consisting of −H, −CH 3 , and –C(=O)‐CH 3 ; R 22 is selected from the group consisting of R 23 is selected from the group consisting of −H, −CH 3 , and −COOH; R 24 is selected from the group consisting of −CH 3 and –C(=O)‐O‐C(CH 3 ) 3 ; R 25 is selected from the group consisting of −NH− an d R 26 is selected from the group consisting of −H and OH; R 27 is selected from the group consisting of −H, −CH 3 , −CH 2 ‐CH 3 , −CN, −CH 2 OH, cyclopropyl, −CH 2 ‐CN, −N(CH 3 ) 2 , −C(CH 3 ) 2 OH, −NH‐C(=O)‐CH 3 , −S(=O) 2 CH 3 , −CH 2 ‐CH 2 ‐OR 36 , −CH 2 ‐ CF 2 R 4a , −C(=O)‐N(R 3a ) 2 , oxetane‐3‐carbonyl, −C(=O)‐C(R 38 )(R 39 )R 4a , and R 28 is selected from the group consisting of −H, −CH 3 , −OH, −N(CH 3 ) 2 , −S(=O) 2 NH 2 , and – C(=O)‐NHR 3a ; R 29 is selected from the group consisting of −H, −CH 3 , and −CH 2 OH; R 31 is selected from the group consisting of −CH 3 and –C(=O)‐CH 3 ; R 32 is selected from the group consisting of −H and CF 3 ; R 33 is selected from the group consisting of −H, −CN , and −CF 3 ; R 34 is selected from the group consisting of −CH 3 and cyclopropyl; R 35 is selected from the group consisting of −CH 3 , −OCH 3 , cyclopropyl, −CH 2 ‐OCH 3 , −CHF 2 , oxetan‐3‐yl, and 1‐methylazetidin‐3‐yl; R 36 is selected from the group consisting of −H, −CH 3 , and R 37 is selected from the group consisting of −H, −F, and −CN; R 38 is selected from the group consisting of −H, −CH 3 , −CH 2 ‐NH 2 , −CH 2 ‐NH‐CH 3 , and −CH 2 ‐ CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐C(=O)‐OR 3a ; and R 39 is selected from the group consisting of −H, −NH 2 , −F, −NH‐CH 3 , −OCH 3 , and −N(CH 3 ) 2 . or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 2. The compound according to claim 1 of general for mula (I) in which: R 1a is selected from the group consisting of 5‐chloro‐1,3‐thiazol‐2‐yl, 6‐aminopyridin‐2‐yl, 5‐bromopyridin‐3‐yl, 3‐(trifluoromethyl)‐1,2,4‐oxadiazol‐5‐yl, 3‐fluoro‐1‐benzofuran‐7‐yl, R 4a is selected from the group consisting of −H and F; R 5a is selected from the group consisting of −H, −F, −Cl, −Br, −CN, −NO 2 , −OH, −CH 2 OH, −COOH, ‐COO‐CH 3 , −CH 3 , −CF 3 , −CHF 2 , −CF 2 −CH 3 , −CF 2 −CH 2 OH, −CF 2 −C(CH 3 ) 2 OH, −O‐CH 3 , −O‐CH 2 ‐CHF 2 , −O‐CF 3 , ‐O‐CHF 2 , −S‐CF 3 , −SF 5 , (1E)‐2‐ethoxyethenyl, and [(tert‐ butoxy)carbonyl]amino; R 6a is selected from the group consisting of −H, −F, −Cl, −CH 3 , −CHF 2 , −O‐CH 3 , −O‐CHF 2 , 1H‐ pyrazol‐1‐yl, 2‐(methylamino)ethoxy, oxolan‐3‐yl oxy, and (1‐methylpyrrolidin‐3‐yl)oxy; R 7 is selected from the group consisting of −H, −NH 2 , −F, and −Br; and R 8 is selected from the group consisting of −H, −CH 3 , and −F; or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 3. The compound according to claim 2 of general for mula (I) in which: R 1a is R 5a is selected from the group consisting of −CF 3 , −CHF 2 , −CF 2 −CH 3 , −CF 2 −CH 2 OH, and −CF 2 −C(CH 3 ) 2 OH; and R 6a is selected from the group consisting of −H, −F, and −CH 3 ; or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 4. The compound according to claim 1 of general for mula (1a) or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 5. The compound according to claim 1 of general for mula (1b) or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 6. The compound according to claim 4 of general for mula (1a) in which: R 1a is R 2a is selected from the group consisting of −F, −Cl , −OCH 3 , −COOCH 3 , −S(=O) 2 ‐CH 3 , −O‐CH 2 ‐ CH 2 R 9 , −C(=O)‐NHR 3a , 2,5‐dihydrofuran‐3‐yl, 4,5‐dihydrofuran‐ 2‐yl, oxolan‐3‐yl, oxetan‐ 3‐yloxy, cyclopentylamino, 5,6‐dihydro‐2H‐pyran‐ 3‐yl, oxan‐3‐yl, 3,6‐dihydro‐2H‐pyran‐ 4‐yl, 1‐methyl‐1H‐pyrazol‐4‐yl, oxan‐4‐yl, [(oxetan‐2‐yl)methyl]amino, −N(R 3a )‐CH(R 3a )‐ CH 2 ‐R 12 , 1‐methylpiperidin‐4‐yl, 1‐methyl‐1,2,3,6 ‐tetrahydropyridin‐4‐yl, 1‐oxa‐6‐ azaspiro[3.3]heptan‐6‐yl, [(oxolan‐2‐yl)methyl]amin o, 2‐oxa‐6‐azaspiro[3.3]heptan‐6‐yl, (1‐methylpyrrolidin‐3‐yl)oxy, (5‐oxopyrrolidin‐3 yl)amino, 3‐(difluoromethyl)azetidin‐1‐ yl, 2‐oxa‐6‐azaspiro[3.4]octan‐6‐yl, 3‐oxo‐1 ,4‐diazepan‐1‐yl, −R 22 ‐COOC(CH 3 ) 3 , 4‐cyano‐4‐ methylpiperidin‐1‐yl, 2‐oxa‐6‐azaspiro[3.5]nonan 6‐yl, 2‐oxa‐7‐azaspiro[3.5]nonan‐7‐yl, 5‐oxo‐2,6‐diazaspiro[3.4]octan‐2‐yl, 7‐oxo‐2, 6‐diazaspiro[3.4]octan‐2‐yl, 8‐oxo‐2,7‐ diazaspiro[4.4]nonan‐2‐yl, 4‐methyl‐2,3‐dioxo‐1 ,4‐diazepan‐1‐yl, , and R 3a is selected from the group consisting of −H and CH 3 ; R 4a is selected from the group consisting of −H and F; R 5a is selected from the group consisting of −CF 3 , −CHF 2 , −CF 2 −CH 3 , −CF 2 −CH 2 OH, and −CF 2 −C(CH 3 ) 2 OH; and R 6a is selected from the group consisting of −H, −F, and −CH 3 ; R 9 is selected from the group consisting of −H, −CH 2 ‐CH 3 , and –NH‐CH 3 ; R 10 is selected from the group consisting of 1 1 R is selected from the group consisting of ‐CH 2 ‐CH 2 ‐CH 2 ‐, ‐CH 2 ‐O‐CH 2 ‐, ‐CH 2 ‐CH 2 ‐O‐, −N(CH 3 )‐CH 2 ‐CH 2 −, ‐CH 2 ‐NH‐CH 2 ‐ and ‐CH 2 ‐N(R 31 )‐CH 2 ‐; R 12 is selected from the group consisting of ‐H, ‐OC H 3 , and ‐N(CH 3 ) 2 ; R 13 is selected from the group consisting of R 14 is selected from the group consisting of −CH 2 ‐C(R 4a ) 2 ‐CH 2 ‐, ‐CH 2 ‐CH 2 ‐C(=O)‐ and ‐CH 2 ‐O‐ C(=O)‐; R 15 is selected from the group consisting of −H, −OH , −F, −OCH 3 , −N(CH 3 ) 2 , −C(=O)‐NH 2 , ‐ COOH, pyrrolidin‐1‐yl, ‐NH‐SO 2 ‐R 34 , ‐N(R 3a )‐CO‐R 35 , and morpholin‐4‐yl; R 16 is selected from the group consisting of −H, −CH 3 , −F, and −CH 2 ‐CH 2 OH; R 17 is selected from the group consisting of −H and N(CH 3 ) 2 ; R 18 is selected from the group consisting of −H and CH=CH 2 ; R 19 is selected from the group consisting of =CH 2 and =O; R 20 is selected from the group consisting of −H and CN; R 21 is selected from the group consisting of −H, −CH 3 , and –C(=O)‐CH 3 ;
R 22 is selected from the group consisting of , , , , , , and ; R 23 is selected from the group consisting of −H, −CH 3 , and −COOH; R 24 is selected from the group consisting of −CH 3 and –C(=O)‐O‐C(CH 3 ) 3 ; R 25 is selected from the group consisting of −NH− an d ; R 26 is selected from the group consisting of −H and OH; R 27 is selected from the group consisting of −H, −CH 3 , −CH 2 ‐CH 3 , −CN, −CH 2 OH, cyclopropyl, −CH 2 ‐CN, −N(CH 3 ) 2 , −C(CH 3 ) 2 OH, −NH‐C(=O)‐CH 3 , −S(=O) 2 CH 3 , −CH 2 ‐CH 2 ‐OR 36 , −CH 2 ‐ CF 2 R 4a , −C(=O)‐N(R 3a ) 2 , oxetane‐3‐carbonyl, −C(=O)‐C(R 38 )(R 39 )R 4a , and ; R 28 is selected from the group consisting of −H, −CH 3 , −OH, −N(CH 3 ) 2 , −S(=O) 2 NH 2 , and – C(=O)‐NHR 3a ; R 29 is selected from the group consisting of −H, −CH 3 , and −CH 2 OH; R 31 is selected from the group consisting of −CH 3 and –C(=O)‐CH 3 ; R 32 is selected from the group consisting of −H and CF 3 ; R 33 is selected from the group consisting of −H, −CN , and −CF 3 ; R 34 is selected from the group consisting of −CH 3 and cyclopropyl; R 35 is selected from the group consisting of −CH 3 , −OCH 3 , cyclopropyl, −CH 2 ‐OCH 3 , −CHF 2 , oxetan‐3‐yl, and 1‐methylazetidin‐3‐yl; R 36 is selected from the group consisting of −H, −CH 3 , and ; R 37 is selected from the group consisting of −H, −F, and −CN; R 38 is selected from the group consisting of −H, −CH 3 , −CH 2 ‐NH 2 , −CH 2 ‐NH‐CH 3 , and −CH 2 ‐ CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐C(=O)‐OR 3a ; and R 39 is selected from the group consisting of −H, −NH 2 , −F, −NH‐CH 3 , −OCH 3 , and −N(CH 3 ) 2 . or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 7. The compound according to claim 5 of general for mula (1b) in which: R 1a is ; R 2a is selected from the group consisting of −F, −Cl , −OCH 3 , −COOCH 3 , −S(=O) 2 ‐CH 3 , −O‐CH 2 ‐ CH 2 R 9 , −C(=O)‐NHR 3a , 2,5‐dihydrofuran‐3‐yl, 4,5‐dihydrofuran‐ 2‐yl, oxolan‐3‐yl, oxetan‐ 3‐yloxy, cyclopentylamino, 5,6‐dihydro‐2H‐pyran‐ 3‐yl, oxan‐3‐yl, 3,6‐dihydro‐2H‐pyran‐ 4‐yl, 1‐methyl‐1H‐pyrazol‐4‐yl, oxan‐4‐yl, [(oxetan‐2‐yl)methyl]amino, −N(R 3a )‐CH(R 3a )‐ CH 2 ‐R 12 , 1‐methylpiperidin‐4‐yl, 1‐methyl‐1,2,3,6 ‐tetrahydropyridin‐4‐yl, 1‐oxa‐6‐ azaspiro[3.3]heptan‐6‐yl, [(oxolan‐2‐yl)methyl]amin o, 2‐oxa‐6‐azaspiro[3.3]heptan‐6‐yl, (1‐methylpyrrolidin‐3‐yl)oxy, (5‐oxopyrrolidin‐3 yl)amino, 3‐(difluoromethyl)azetidin‐1‐ yl, 2‐oxa‐6‐azaspiro[3.4]octan‐6‐yl, 3‐oxo‐1 ,4‐diazepan‐1‐yl, −R 22 ‐COOC(CH 3 ) 3 , 4‐cyano‐4‐ methylpiperidin‐1‐yl, 2‐oxa‐6‐azaspiro[3.5]nonan 6‐yl, 2‐oxa‐7‐azaspiro[3.5]nonan‐7‐yl, 5‐oxo‐2,6‐diazaspiro[3.4]octan‐2‐yl, 7‐oxo‐2, 6‐diazaspiro[3.4]octan‐2‐yl, 8‐oxo‐2,7‐ diazaspiro[4.4]nonan‐2‐yl, 4‐methyl‐2,3‐dioxo‐1 ,4‐diazepan‐1‐yl,
, , , , , and ; R 3a is selected from the group consisting of −H and CH 3 ; R 4a is selected from the group consisting of −H and F; R 5a is selected from the group consisting of −CF 3 , −CHF 2 , −CF 2 −CH 3 , −CF 2 −CH 2 OH, and −CF 2 −C(CH 3 ) 2 OH; and R 6a is selected from the group consisting of −H, −F, and −CH 3 ; R 9 is selected from the group consisting of −H, −CH 2 ‐CH 3 , and –NH‐CH 3 ; R 10 is selected from the group consisting of and ; R 11 is selected from the group consisting of ‐CH 2 ‐CH 2 ‐CH 2 ‐, ‐CH 2 ‐O‐CH 2 ‐, ‐CH 2 ‐CH 2 ‐O‐, −N(CH 3 )‐CH 2 ‐CH 2 −, ‐CH 2 ‐NH‐CH 2 ‐ and ‐CH 2 ‐N(R 31 )‐CH 2 ‐; R 12 is selected from the group consisting of ‐H, ‐OC H 3 , and ‐N(CH 3 ) 2 ; R 13 is selected from the group consisting of , , , , and ; R 14 is selected from the group consisting of −CH 2 ‐C(R 4a ) 2 ‐CH 2 ‐, ‐CH 2 ‐CH 2 ‐C(=O)‐ and ‐CH 2 ‐O‐ C(=O)‐; R 15 is selected from the group consisting of −H, −OH , −F, −OCH 3 , −N(CH 3 ) 2 , −C(=O)‐NH 2 , ‐ COOH, pyrrolidin‐1‐yl, ‐NH‐SO 2 ‐R 34 , ‐N(R 3a )‐CO‐R 35 , and morpholin‐4‐yl; R 16 is selected from the group consisting of −H, −CH 3 , −F, and −CH 2 ‐CH 2 OH; R 17 is selected from the group consisting of −H and N(CH 3 ) 2 ; R 18 is selected from the group consisting of −H and CH=CH 2 ; R 19 is selected from the group consisting of =CH 2 and =O; R 20 is selected from the group consisting of −H and CN; R 21 is selected from the group consisting of −H, −CH 3 , and –C(=O)‐CH 3 ;
R 22 is selected from the group consisting of , , , , , , and ; R 23 is selected from the group consisting of −H, −CH 3 , and −COOH; R 24 is selected from the group consisting of −CH 3 and –C(=O)‐O‐C(CH 3 ) 3 ; R 25 is selected from the group consisting of −NH− an d ; R 26 is selected from the group consisting of −H and OH; R 27 is selected from the group consisting of −H, −CH 3 , −CH 2 ‐CH 3 , −CN, −CH 2 OH, cyclopropyl, −CH 2 ‐CN, −N(CH 3 ) 2 , −C(CH 3 ) 2 OH, −NH‐C(=O)‐CH 3 , −S(=O) 2 CH 3 , −CH 2 ‐CH 2 ‐OR 36 , −CH 2 ‐ CF 2 R 4a , −C(=O)‐N(R 3a ) 2 , oxetane‐3‐carbonyl, −C(=O)‐C(R 38 )(R 39 )R 4a , and ; R 28 is selected from the group consisting of −H, −CH 3 , −OH, −N(CH 3 ) 2 , −S(=O) 2 NH 2 , and – C(=O)‐NHR 3a ; R 29 is selected from the group consisting of −H, −CH 3 , and −CH 2 OH; R 31 is selected from the group consisting of −CH 3 and –C(=O)‐CH 3 ; R 32 is selected from the group consisting of −H and CF 3 ; R 33 is selected from the group consisting of −H, −CN , and −CF 3 ; R 34 is selected from the group consisting of −CH 3 and cyclopropyl; R 35 is selected from the group consisting of −CH 3 , −OCH 3 , cyclopropyl, −CH 2 ‐OCH 3 , −CHF 2 , oxetan‐3‐yl, and 1‐methylazetidin‐3‐yl; R 36 is selected from the group consisting of −H, −CH 3 , and ; R 37 is selected from the group consisting of −H, −F, and −CN;
R 38 is selected from the group consisting of −H, −CH 3 , −CH 2 ‐NH 2 , −CH 2 ‐NH‐CH 3 , and −CH 2 ‐ CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐CH 2 ‐C(=O)‐OR 3a ; and R 39 is selected from the group consisting of −H, −NH 2 , −F, −NH‐CH 3 , −OCH 3 , and −N(CH 3 ) 2 . or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 8. A compound according to claim 1, which is select ed from the group consisting of: N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐fluoro‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoro‐phenyl] ethyl]‐2‐methyl‐6‐pyrrolidin‐1‐yl‐pyrido[3,4 d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]eth yl}‐6‐fluoro‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(1R)‐1‐[3‐(difluoromethyl)‐2‐methyl‐phenyl] ethyl]‐2‐methyl‐6‐pyrrolidin‐1‐yl‐pyrido[3,4 d]pyrimidin‐4‐amine 6‐fluoro‐2‐methyl‐N‐[(1R)‐1‐[3‐(trifluoromet hyl)phenyl]ethyl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐[(3R)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluo romethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluo romethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl]‐6 fluoro‐2‐methyl‐pyrido[3,4‐d]pyrimidin‐4‐ amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide
N‐[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl] 6‐fluoro‐2‐methyl‐pyrido[3,4‐d]pyrimidin‐4‐ amine N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) ‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3, 4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) ‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3, 4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐fluoro‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2,8‐dimethylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2,8‐dimethyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one N‐{(3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)pheny l]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3S)‐1‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl 3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide 6‐ethoxy‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromet hyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 1‐(3‐{(1R)‐1‐[(6‐ethoxy‐2‐methylpyrido[3,4‐d ]pyrimidin‐4‐yl)amino]ethyl}‐2‐fluorophenyl)‐1,1 difluoro‐2‐methylpropan‐2‐ol 6‐ethoxy‐N‐{(1R)‐1‐[2‐fluoro‐3‐(trifluoromet hyl)phenyl]ethyl}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluoropheny l]ethyl}‐6‐ethoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]eth yl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine 6‐ethoxy‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}‐6‐eth oxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine
N‐{(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phenyl] ethyl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine 6‐methoxy‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluorome thyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐ amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]et hyl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]eth yl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine 6‐methoxy‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3 (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluoropheny l]ethyl}‐6‐methoxy‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine 2,2‐difluoro‐2‐(2‐fluoro‐3‐{(1R)‐1‐[(6‐met hoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl)amino]ethyl}phenyl)ethan‐1‐ol 1,1‐difluoro‐1‐(2‐fluoro‐3‐{(1R)‐1‐[(6‐met hoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl)amino]ethyl}phenyl)‐2‐methylpropan‐2‐ol N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2 hydroxyethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl} acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2 hydroxy‐2‐methylpropyl)‐2‐fluorophenyl]ethyl}amino) ‐ 2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3 yl}acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[2‐fluoro‐3‐(trifluo romethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl 3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)pheny l]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(2‐methylph enyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(3‐methylph enyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(4‐methylph enyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide
N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2‐fluorophenyl)ethyl ]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐fluorophenyl)ethyl]am ino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(4‐fluorophenyl)ethyl]am ino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2‐methoxyphenyl)ethyl]a mino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐methoxyphenyl)ethyl]a mino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2‐chlorophenyl)ethyl]am ino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐chlorophenyl)ethyl]am ino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[4‐({(1RS)‐1‐[2‐(difluoromethyl)phen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1RS)‐1‐[2‐(difluoromethoxy)phe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethoxy)phen yl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1R)‐1‐[3‐(trifluo romethoxy)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromophenyl)ethyl]ami no}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐{3‐[(triflu oromethyl)sulfanyl]phenyl}ethyl]amino}pyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1R)‐1‐[3‐(pentafl uoro‐lambda 6 ‐ sulfanyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐y l]pyrrolidin‐3‐yl}acetamide methyl 3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidi n‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]benzoate N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐cyanophenyl)ethyl]ami no}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(2‐methyl‐4‐{[(1R)‐1‐(3‐nitrophe nyl)ethyl]amino}pyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide
tert‐butyl {3‐[(1RS)‐1‐({6‐[(3R)‐3‐acetami dopyrrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin 4‐ yl}amino)ethyl]phenyl}carbamate N‐[(3R)‐1‐(4‐{[(1R)‐1‐(4‐fluoro‐3‐methylph enyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6 yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,3‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3,4‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,4‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(3,5‐difluorophenyl)eth yl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(2,6‐difluorophenyl)eth yl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(2,5‐difluorophenyl)eth yl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromo‐2‐methylphe nyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromo‐5‐fluorophe nyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromo‐4‐fluorophe nyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐bromo‐2‐fluorophe nyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromo‐2‐fluorophe nyl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromo‐2‐methoxyph enyl)ethyl]amino}‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐fluoro‐1‐benzofur an‐7‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide. N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3‐fluoro‐1‐benzofur an‐7‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidi n‐ 6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1RS)‐1‐[2‐(1H‐p yrazol‐1‐yl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide
N‐{(3R)‐1‐[4‐({(1RS)‐1‐[3‐(difluoromethyl) 1‐methyl‐1H‐pyrazol‐4‐yl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl} acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1RS)‐1‐[1‐methyl 3‐(trifluoromethyl)‐1H‐pyrazol‐4‐ yl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin 3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1RS)‐1‐(5‐chloro‐1,3‐thiaz ol‐2‐yl)ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidi n‐ 6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[2‐methyl‐4‐({(1RS)‐1‐[3‐(triflu oromethyl)‐1,2,4‐oxadiazol‐5‐ yl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin 3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(5‐bromopyridin‐3‐yl )ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(6‐aminopyridin‐2‐yl )ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐amino‐5‐(trifluor omethyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[1‐(3‐aminophenyl)ethyl]amino}‐2 ‐methylpyrido[3,4‐d]pyrimidin‐6‐yl)pyrrolidin‐ 3‐yl]acetamide (mixture of stereoisomers) tert‐butyl {3‐[(1S)‐1‐({6‐[(3R)‐3‐acetamidopy rrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4 yl}amino)ethyl]phenyl}carbamate tert‐butyl {3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopy rrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4 yl}amino)ethyl]phenyl}carbamate N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3‐aminophenyl)ethyl]ami no}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐aminophenyl)ethyl]ami no}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3,5‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3,5‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1S)‐1‐(2,6‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,6‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,5‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide
N‐[(3R)‐1‐(4‐{[(1S)‐1‐(2,5‐difluorophenyl)e thyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide 3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidin‐1‐ yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]benzoic acid N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(hydroxymethyl)phenyl ]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐hydroxyphenyl)ethyl]a mino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(2,2‐difluoroethoxy )phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐[(3R)‐1‐(4‐{[(1R)‐1‐{3‐[(E)‐2‐ethoxyethe nyl]phenyl}ethyl]amino}‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl)pyrrolidin‐3‐yl]acetamide N‐{(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}‐2‐met hyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]eth yl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl}‐2 methyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluoropheny l]ethyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]et hyl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine 2,2‐difluoro‐2‐{2‐fluoro‐3‐[(1R)‐1‐{[2‐met hyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐d]pyrim idin‐ 4‐yl]amino}ethyl]phenyl}ethan‐1‐ol 1,1‐difluoro‐1‐{2‐fluoro‐3‐[(1R)‐1‐{[2‐met hyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4‐d]pyrim idin‐ 4‐yl]amino}ethyl]phenyl}‐2‐methylpropan‐2‐ol N‐{(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phenyl]eth yl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}‐6‐[(3 R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl}‐6 [(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluoropheny l]ethyl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐ yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine
6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐N {(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phenyl]ethyl }‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]eth yl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine 2‐{3‐[(1R)‐1‐({6‐[(3R)‐3‐(dimethylamino)pyrrol idin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]‐2‐fluorophenyl}‐2,2‐difluoroethan‐ 1‐ol 1‐{3‐[(1R)‐1‐({6‐[(3R)‐3‐(dimethylamino)pyrrol idin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]‐2‐fluorophenyl}‐1,1‐difluoro‐2‐m ethylpropan‐2‐ol 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl}amin o)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐2,6‐ diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphen yl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl }amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐ 2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[2‐fluoro‐3‐(trifluoromethyl)phe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2‐hydroxyethy l)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2‐[4‐({(1R)‐1‐[3‐amino‐5‐(trifluoromethyl)phen yl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)phenyl]ethyl }amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl] ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐ fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐methy lphenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[2‐fluoro‐3‐(trifluoromethy l)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(1,1‐difluoro‐2‐hydrox y‐2‐methylpropyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]piperazin‐1‐yl}e than‐1‐one
1‐{4‐[4‐({(1R)‐1‐[3‐amino‐5‐(trifluoromet hyl)phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ‐ethyl‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 6 ‐cyclopropyl‐N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methyl‐N 6 ‐(propan‐2‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ‐ethyl‐N 6 ,2‐dimethylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ,2‐dimethyl‐N 6 ‐(prop‐2‐en‐1‐ yl)pyrido[3,4‐d]pyrimidine‐4,6‐diamine N 6 ‐cyclopropyl‐N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ,2‐dimethylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 6 ‐cyclobutyl‐N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ,2‐dimethyl‐N 6 ‐(propan‐2‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ‐(2‐methoxyethyl)‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(piperidin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N 6 ‐cyclopentyl‐N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(piperazin‐1‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐ol (3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐ol N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(morpholin‐4‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methyl‐N 6 ‐{[(2RS)‐oxetan‐2‐ yl]methyl}pyrido[3,4‐d]pyrimidine‐4,6‐diamine
N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methyl‐N 6 ‐[(3R)‐oxolan‐3‐yl]pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ‐(2‐methoxyethyl)‐N 6 ,2‐ dimethylpyrido[3,4‐d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methyl‐N 6 ,N 6 ‐di(prop‐2‐en‐1‐ yl)pyrido[3,4‐d]pyrimidine‐4,6‐diamine 6‐[2‐azabicyclo[2.2.1]heptan‐2‐yl]‐N‐{(1R)‐1 [3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of st ereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(1‐oxa‐6‐azaspiro[3.3]heptan‐ 6‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(2‐oxa‐6‐azaspiro[3.3]heptan‐ 6‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine N 6 ‐cyclohexyl‐N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine 4‐{[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]amino}pyrrolidin‐2‐one (mixture o f stereoisomers) 4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]piperazin‐2‐one 6‐(1,4‐diazepan‐1‐yl)‐N‐{(1R)‐1‐[3‐(difluo romethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4 d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(3R)‐3‐methylmorpholin‐4‐ yl]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(3S)‐3‐methylmorpholin‐4‐ yl]pyrido[3,4‐d]pyrimidin‐4‐amine (3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐3‐ol (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐3‐ol N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methyl‐N 6 ‐(oxan‐4‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐2‐methyl‐N 6 ‐{[(2R)‐oxolan‐2‐ yl]methyl}pyrido[3,4‐d]pyrimidine‐4,6‐diamine
N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl] ethyl}‐6‐[(3S)‐3‐methoxypyrrolidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ‐[2‐(dimethylamino)ethyl]‐N 6 ,2‐ dimethylpyrido[3,4‐d]pyrimidine‐4,6‐diamine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(thiomorpholin‐4‐yl)pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐[3‐(difluoromethyl)azetidin‐1‐yl]‐N‐{(1R)‐1 [3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐(3,3‐difluoropyrrolidin‐1‐yl)‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐(2,6‐dihydropyrrolo[3,4‐c]pyrazol‐5(4H)‐ yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]piperidine‐4‐carbonitrile N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[hexahydrocyclopenta[c]pyrrol‐2(1H)‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[hexahydropyrrolo[3,4‐c]pyrrol‐2(1H)‐ yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixtu re of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(3aR,6aS)‐tetrahydro‐1H‐ furo[3,4‐c]pyrrol‐5(3H)‐yl]pyrido[3,4‐d]pyrimidin‐ 4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(3aRS,6aRS)‐hexahydro‐5H‐furo[2,3‐ c]pyrrol‐5‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4 amine (mixture of stereisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(2‐oxa‐6‐azaspiro[3.4]octan‐6 ‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N 6 ‐cyclohexyl‐N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ,2‐dimethylpyrido[3,4‐ d]pyrimidine‐4,6‐diamine 4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐1,4‐diazepan‐2‐one (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidine‐3‐carboxamide (6R)‐4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐6‐methylpiperazin‐2‐one (6S)‐4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐6‐methylpiperazin‐2‐one
N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl] ethyl}‐6‐(3,3‐dimethylpiperazin‐1‐yl)‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(4‐methyl‐1,4‐diazepan‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐(4‐ethylpiperazin‐1‐yl)‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(3S)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine {1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐4‐yl}methanol N 4 ‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoropheny l]ethyl}‐N 6 ,2‐dimethyl‐N 6 ‐(oxan‐4‐yl)pyrido[3,4‐ d]pyrimidine‐4,6‐diamine 4‐{[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]amino}cyclohexan‐1‐ol (mixture of stereoisomers) (1RS,4SR,5RS)‐2‐[4‐({(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐2‐azabicyclo[2. 2.1]heptane‐5‐carbonitrile (mixture of stereoisomers) N 2 ‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐N,N,N 2 ‐trimethylglycinamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐(6,7‐dihydropyrazolo[1,5‐a]pyrazin‐ 5(4H)‐yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐(5,6‐dihydroimidazo[1,5‐a]pyrazin‐7(8H)‐ yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐(5,6‐dihydroimidazo[1,2‐a]pyrazin‐7(8H)‐ yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(1‐methyl‐4,6‐ dihydropyrrolo[3,4‐c]pyrazol‐5(1H)‐yl)pyrido[3,4‐d]p yrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐(5,6‐dihydro[1,2,4]triazolo[1,5‐a]pyrazin‐ 7(8H)‐yl)‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐4‐methylpiperidine‐4‐carbonitrile {4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}acetonitrile
2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorop henyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐2,6‐diazaspiro[3.4]octan‐5‐one 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(3aS,6aS)‐1‐ methylhexahydropyrrolo[3,4‐b]pyrrol‐5(1H)‐yl]pyrido[3, 4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(3aRS,6aSR)‐5‐ methylhexahydropyrrolo[3,4‐c]pyrrol‐2(1H)‐yl]pyrido[3, 4‐d]pyrimidin‐4‐amine (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(3aR,6aR)‐1‐ methylhexahydropyrrolo[3,4‐b]pyrrol‐5(1H)‐yl]pyrido[3, 4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(8aS)‐hexahydropyrrolo[1,2‐a]pyrazin‐ 2(1H)‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(8aR)‐hexahydropyrrolo[1,2‐a]pyrazin‐ 2(1H)‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(6‐methyl‐2,6‐ diazaspiro[3.4]octan‐2‐yl)pyrido[3,4‐d]pyrimidin‐4 amine 6‐(4‐cyclopropylpiperazin‐1‐yl)‐N‐{(1R)‐1‐[3 ‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(2‐oxa‐6‐azaspiro[3.5]nonan‐6 ‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(2‐oxa‐7‐azaspiro[3.5]nonan‐7 ‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine (3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fl uorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐3‐methylpyrrolidine‐3‐carboxam ide 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]piperidine‐4‐carboxamide 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one (3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐3‐carboxamide (3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐3‐carboxamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(cis)‐3,4,5‐trimethylpiperazin 1‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine (mixture of s tereoisomers)
N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl] ethyl}‐2‐methyl‐6‐[(3R,5R)‐3,4,5‐ trimethylpiperazin‐1‐yl]pyrido[3,4‐d]pyrimidin‐4‐a mine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(3S,5S)‐3,4,5‐ trimethylpiperazin‐1‐yl]pyrido[3,4‐d]pyrimidin‐4‐a mine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[3‐(dimethylamino)piperidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of st ereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[4‐(dimethylamino)piperidin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐3‐methylpyrrolidine‐3‐carboxylic acid (mix ture of stereoisomers) 4‐{[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]amino}‐1‐methylcyclohexan‐1‐ol (mixture of stereoisomers) 2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐ol 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐3‐(2‐hydroxyethyl)pyrrolidin‐3‐ol (mixtur e of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(3‐methyl‐5,6‐ dihydro[1,2,4]triazolo[4,3‐a]pyrazin‐7(8H)‐yl)pyrido[3 ,4‐d]pyrimidin‐4‐amine 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]hexahydropyrrolo[1,2‐a]pyrazin‐6(2H)‐one (mixt ure of stereoisomers) (5RS)‐7‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fl uorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,7‐diazaspiro[4.4]nonan‐3‐one (mixture of stereoisomers) 6‐[[1,3'‐bipyrrolidin]‐1'‐yl]‐N‐{(1R)‐1‐[3 (difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine (mixture of st ereoisomers) 7‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]hexahydro‐3H‐[1,3]oxazolo[3,4‐a]pyrazin‐3‐o ne (mixture of stereoisomers) 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐4‐methyl‐1,4‐diazepane‐2,3‐dione 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐1,4‐diazepan‐1‐yl}ethan‐1‐ one N‐{(3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 ‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐N‐methylacetam ide (mixture of stereoisomers) N‐{1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐4‐yl}acetamide
(3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐N‐methylpiperidine‐3‐carboxami de (mixture of stereoisomers) 2‐{1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidin‐4‐yl}propan‐2‐ol (2R)‐4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐6‐oxopiperazine‐2‐carboxylic acid N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[4‐(2‐methoxyethyl)piperazin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine 5‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐4,5,6,7‐tetrahydropyrazolo[1,5‐a]pyrazine‐2 carbonitrile 6‐[4‐(2,2‐difluoroethyl)piperazin‐1‐yl]‐N‐{(1R )‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2 methylpyrido[3,4‐d]pyrimidin‐4‐amine 1‐[5‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]hexahydropyrrolo[3,4‐c]pyrrol‐2(1H) ‐yl]ethan‐1‐one (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[3‐(piperidin‐1‐yl)pyrrolidin 1‐ yl]pyrido[3,4‐d]pyrimidin‐4‐amine (mixture of stere oisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[3‐(morpholin‐4‐yl)pyrrolidin 1‐yl]pyrido[3,4‐d]pyrimidin‐4‐amine (mixture of s tereoisomers) 6‐[7,7‐difluorohexahydropyrrolo[1,2‐a]pyrazin‐2(1H) yl]‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐ fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐d]pyrimidin‐4 ‐amine (mixture of stereoisomers) (3RS)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fl uorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐3‐sulfonamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[4‐(2,2,2‐ trifluoroethyl)piperazin‐1‐yl]pyrido[3,4‐d]pyrimidin 4‐amine tert‐butyl {(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluorome thyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl} carbamate tert‐butyl {3‐[4‐({(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐3‐azabicyclo[3.1.0]hexan‐1‐yl} carbamate (mixture of stereoisomers) tert‐butyl {1‐[4‐({(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐4‐fluoropyrrolidin‐3‐yl}carbam ate (mixture of stereoisomers) tert‐butyl 6‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐ 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octane‐2‐ca rboxylate tert‐butyl 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐ 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,7‐diazaspiro[3.5]nonane‐7‐ca rboxylate
tert‐butyl 7‐[4‐({(1R)‐1‐[3‐(difluoromethyl) ‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,7‐diazaspiro[3.5]nonane‐2‐ca rboxylate N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐(6‐methyl‐2,6‐ diazaspiro[3.4]octan‐2‐yl)pyrido[3,4‐d]pyrimidin‐4 amine tert‐butyl 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐ 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octane‐6‐ca rboxylate methyl 4‐(2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl )‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]piperazin‐1‐yl}e thoxy)benzoate 4‐(2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐ fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethoxy)benzoic acid 6‐(methanesulfonyl)‐2‐methyl‐N‐{(1R)‐1‐[3‐(t rifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐[(3R)‐3‐aminopyrrolidin‐1‐yl]‐N‐{(1R)‐1‐ [3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride s alt N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}cyclopropanecarboxa mide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐2,2‐difluoroac etamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐2‐methoxyaceta mide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}oxetane‐3‐carbo xamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐1‐methylazetid ine‐3‐carboxamide methyl {(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl} carbamate N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}methanesulfonamide N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}cyclopropanesulfona mide cyclopropyl{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}methanone 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2‐methoxyethan 1‐one
1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fl uorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2,2‐difluoroeth an‐1‐one {4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}(oxetan‐3‐yl)met hanone 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2‐(dimethylamin o)ethan‐1‐one {4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}(1‐fluorocycloprop yl)methanone 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2,2‐difluoropro pan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazine‐1‐carbonyl}cyclopropane 1‐carbonitrile methyl 10‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐ 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐10‐oxodecanoate 10‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluo rophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐10‐oxodecanoic acid 4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐yl]‐N,N‐dimethylpiperazine‐1‐carboxamide N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[4‐(methanesulfonyl)piperazin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine 2‐amino‐1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl) ‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐2‐(methylamino) ethan‐1‐one 3‐amino‐1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl) ‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}propan‐1‐one 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}‐3‐(methylamino) propan‐1‐one 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐me thyl‐N‐{(1R)‐1‐[3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine 2‐[2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phe nyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]‐ 2,6‐diazaspiro[3.4]octan‐7‐one 1‐{4‐[2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethy l)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one
2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{(1 R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐fluoro‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐ 4‐amine 2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{(1R) 1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine 2‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(triflu oromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐me thyl‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine 1‐{4‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(t rifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐(trifluoromet hyl)phenyl]ethyl}‐6‐(1‐oxa‐6‐azaspiro[3.3]heptan 6‐yl)pyrido[3,4‐d]pyrimidin‐4‐amine 6‐fluoro‐2,8‐dimethyl‐N‐{(1R)‐1‐[3‐(trifluor omethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐ amine 1‐{4‐[2,8‐dimethyl‐4‐({(1R)‐1‐[3‐(trifluorom ethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐ yl]piperazin‐1‐yl}ethan‐1‐one 2,8‐dimethyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{( 1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2,8‐ dimethyl‐N‐{(1R)‐1‐[3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine 2‐[2,8‐dimethyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl )phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin‐6‐yl]‐ 2,6‐diazaspiro[3.4]octan‐7‐one 6‐fluoro‐2,8‐dimethyl‐N‐{(1R)‐1‐[2‐methyl‐ 3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 1‐{4‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐methyl‐3 (trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one 2‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐methyl‐3‐(tr ifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]‐2,6‐diazaspiro[3.4]octan‐7‐one 2,8‐dimethyl‐6‐(4‐methylpiperazin‐1‐yl)‐N‐{( 1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2,8‐ dimethyl‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine
N‐{(3R)‐1‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐m ethyl‐3‐ (trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidin ‐6‐yl]pyrrolidin‐3‐yl}acetamide N‐{(3S)‐1‐[2,8‐dimethyl‐4‐({(1R)‐1‐[2‐meth yl‐3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide 6‐chloro‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromet hyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine 2‐methyl‐6‐(1‐methyl‐1H‐pyrazol‐4‐yl)‐N‐ {(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐(4,5‐dihydrofuran‐2‐yl)‐2‐methyl‐N‐{(1R) 1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐(2,5‐dihydrofuran‐3‐yl)‐2‐methyl‐N‐{(1R) 1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine 6‐(3,6‐dihydro‐2H‐pyran‐4‐yl)‐2‐methyl‐N {(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4 d]pyrimidin‐4‐amine 6‐(5,6‐dihydro‐2H‐pyran‐3‐yl)‐2‐methyl‐N {(1R)‐1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4 d]pyrimidin‐4‐amine 2‐methyl‐6‐(1‐methyl‐1,2,3,6‐tetrahydropyridin 4‐yl)‐N‐{(1R)‐1‐[3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine 2‐methyl‐6‐[(3RS)‐oxolan‐3‐yl]‐N‐{(1R)‐1 [3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine amine (mixture of stereoisomers) 2‐methyl‐6‐(oxan‐4‐yl)‐N‐{(1R)‐1‐[3‐(tri fluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐ amine 2‐methyl‐6‐[(3RS)‐oxan‐3‐yl]‐N‐{(1R)‐1‐[ 3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine (mixture of stereoisomers) 2‐methyl‐6‐(1‐methylpiperidin‐4‐yl)‐N‐{(1R) 1‐[3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐ d]pyrimidin‐4‐amine methyl 2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl) phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxylate 2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]e thyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxamide N,2‐dimethyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phen yl]ethyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxamide 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphen yl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperidine‐4‐carbonitrile N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(2S)‐2,4‐dimethylpiperazin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine
{1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoro phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]‐4‐methylpiperazin‐2‐yl}methano l (mixture of stereoisomers) N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐ dihydroimidazo[1,2‐a]pyrazin‐7(8H)‐yl]pyrido[3,4‐d]p yrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐ dihydro[1,2,4]triazolo[1,5‐a]pyrazin‐7(8H)‐yl]pyrido[3 ,4‐d]pyrimidin‐4‐amine 6‐(cyclobutyloxy)‐N‐{(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine 6‐butoxy‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}‐2‐methylpyrido[3,4‐d]pyrimidin‐ 4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[2‐ (methylamino)ethoxy]pyrido[3,4‐d]pyrimidin‐4‐amine N‐[(1R)‐1‐{3‐(difluoromethyl)‐2‐[2‐(methylamin o)ethoxy]phenyl}ethyl]‐2‐methyl‐6‐[2‐ (methylamino)ethoxy]pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[(oxetan‐3‐yl)oxy]pyrido[3,4‐ d]pyrimidin‐4‐amine tert‐butyl 3‐{[4‐({(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]oxy}azetidine‐1‐carboxylate N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐{[(3R)‐oxolan‐3‐ yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐{[(3R)‐oxolan 3‐yl]oxy}phenyl]ethyl}‐2‐methyl‐6‐{[(3R)‐oxola n‐ 3‐yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐{[(3S)‐oxolan‐3‐ yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐{[(3S)‐oxolan 3‐yl]oxy}phenyl]ethyl}‐2‐methyl‐6‐{[(3S)‐oxola n‐ 3‐yl]oxy}pyrido[3,4‐d]pyrimidin‐4‐amine N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐{[(3S)‐1‐met hylpyrrolidin‐3‐yl]oxy}phenyl]ethyl}‐2‐methyl‐6‐ {[(3S)‐1‐methylpyrrolidin‐3‐yl]oxy}pyrido[3,4‐d]py rimidin‐4‐amine 6‐[(azetidin‐3‐yl)oxy]‐N‐{(1R)‐1‐[3‐(difluor omethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine hydrochloride tert‐butyl {(3‐trans)‐1‐[4‐({(1R)‐1‐[3‐(dif luoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐4‐fluoropyrroli din‐3‐yl}carbamate (mixture of stereoisomers) 6‐[(trans)‐3‐amino‐4‐fluoropyrrolidin‐1‐yl]‐ N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐ 2‐methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochlorid e (mixture of stereoisomers)
tert‐butyl {(cis)‐1‐[4‐({(1R)‐1‐[3‐(difluo romethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐4‐fluoropyrroli din‐3‐yl}carbamate (mixture of stereoisomers) 6‐[(cis)‐3‐amino‐4‐fluoropyrrolidin‐1‐yl]‐N {(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl }‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride ( mixture of stereoisomers) or a stereoisomer, a tautomer, an N‐oxide, a hydra te, a solvate, or a salt thereof, or a mixture of same. 9. A compound of general formula (1) according to a ny one of claims 1 to 8 for use in the treatment or prophylaxis of a disease. 10. A pharmaceutical composition comprising a compound of general formula (1) according to any one of claims 1 to 8 and one or more pharmaceutica lly acceptable excipients. 11. A pharmaceutical combination comprising: ^ one or more first active ingredients, in particular compounds of general formula (1) according to any one of claims 1 to 8, and ^ one or more further active ingredients, in particular anti‐hyperproliferative and/or anti‐ cancer agents. 12. Use of a compound of general formula (1) accord ing to any one of claims 1 to 8 for the treatment or prophylaxis of a disease. 13. Use of a compound of general formula (1) according to any one of claims 1 to 8 for the preparation of a medicament for the treatment or pro phylaxis of a disease. 14. Use according to claim 9, 12 or 13, wherein th e disease is a hyperproliferative disorder, such as cancer, for example. 15. Use of SOS1 Inhibitors for the treatment or pro phylaxis of a disease, especially for the treatment or prophylaxis of cancer. The present invention covers any sub‐combination wit hin any embodiment or aspect of the present invention of compounds of general formula (I), supra. The present invention covers any sub‐combination wit hin any embodiment or aspect of the present invention of intermediate compounds of general formula (II).The present invention covers the compounds of general formula (I) which are disclosed in the Example Section of this text, infra. SYNTHESIS OF COMPOUNDS (OVERVIEW) The compounds of the present invention can be prepar ed as described in the following section. The schemes and the procedures described below illustrate general synthetic routes to the compounds
of general formula (I) of the invention and are not intended to be limiting. It is clear to the pe rson skilled in the art that the order of transformations as exemplified in the schemes can be modified in various ways. The order of transformations exemplified in the schemes is therefore not intended to be limiting. In addition, interconversion of any of the substituents can be achieved before and/or after the exemplified transformations. These modificati ons can be such as the introduction of protecting groups, cleavage of protecting groups, exch ange, reduction or oxidation of functional groups, halogenation, metallation, substitution or othe r reactions known to the person skilled in the art. These transformations include those which introdu ce a functionality which allows for further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well‐known to the person skilled in the art ( see for example P.G.M. Wuts and T.W. Greene in "Protective Groups in Organic Synthesis", 4'" edition, Wiley 2006). Specific examples are described in the subsequent paragraphs. Further, it is possible th at two or more successive steps may be performed without work‐up being performed between sa id steps, e.g. a "one‐pot" reaction, as is well‐known to the person skilled in the art. The syntheses of the compounds of the present invent ion are preferably carried out according to the general synthetic sequence, shown in schemes 1‐7.
Scheme 1: Route for the preparation of compounds of general formula 8, wherein T, V, R 1 and x have the meaning as given for general formula (I), supra and R is alkyl, Hal is chloro, bromo or iodo and LG has the meaning as a leaving group, preferably chlor o, bromo or a sulfonate group as depicted in scheme 1. Specific examples are described in the sub sequent paragraphs. Step 1 ^ 7 (Scheme 1) Azaquinazoline formation In the first step (scheme 1) amino acid ester deriv ative 1 (which is commercially available or described in the literature) can be converted to the corresponding azaquinazoline 7 in analogy to literature procedures. Typically acetonitrile and hydro chloric acid in organic solvent such as for example 1,4‐dioxane at elevated temperatures is used . For example see ACS Medicinal Chemistry Letters, 2013, vol. 4, # 9 p. 846 – 851; Journal of Medicinal Chemistry, 2009, vol. 52, # 8 p. 234 1 ‐ 2351 or WO2015/54572 and references therein.
Step 2 ^ 7 (Scheme 1) Azaquinazoline formation Alternatively halogen substituted benzoic acid derivati ve of general formula 2 (which is commercially available or described in the literature) can be con verted to the corresponding azaquinazoline 7 in analogy to literature procedures. Typically derivative 2 is reacted with acetamidine, copper metal, a base such as for example potassium carbonate in an organic solvent such as for example DMF at elevated temperature. For example see WO2005/51410, US 2008/107623 and references therein. Step 3 ^ 7 (Scheme 1) Azaquinalzoline formation Alternatively amino substituted benzoic acid derivative of general formula 3 (which is commercially available or described in the literature) can be con verted to the corresponding azaquinazoline 7 in analogy to literature procedures. Typically derivative 3 is reacted with acetyl chloride or acetic anhydride, an ammonia source such as for example amm onia or ammonium acetate, a base such as for example triethylamine or pyridine with or without DMAP in an organic solvent such as for example DMF, toluene, 1,4‐dioxane / water at elevat ed temperature. For example see Bioorganic and Medicinal Chemistry Letters, 2011, vol. 21, # 4 p. 1270 – 1274; Bioorganic and Medicinal Chemistry Letters, 2010, vol. 20, # 7 p. 2330 – 2334; WO2008/117079 or WO2006/74187 and references therein. Step 4 ^ 7 (Scheme 1) Azaquinazoline formation Alternatively benzoxazinone derivative of general formu la 4 (which is commercially available or can be prepared in analogy to literature procedures) can be converted to the corresponding azaquinazoline 7 in analogy to literature procedures. Typically derivative 4 is reacted with ammonium acetate in a solvent at elevated temperature . For example see Bioorganic and Medicinal Chemistry Letters, 2011, vol. 21, # 4 p. 1270 – 1274 or US6350750 and references therein. Step 5 ^ 7 (Scheme 1) Azaquinazoline formation Alternatively benzoic acid amide derivative of general formula 5 (which is commercially available or described in the literature) can be converted to the corresponding azaquinazoline 7 in analogy to literature procedures. Typically derivative 5 is react ed with a base such as for example sodium hydroxide in a solvent such as for example water at elevated temperature. For example see
Bioorganic and Medicinal Chemistry Letters, 2008, v ol. 18, # 16 p. 4573 – 4577 and references therein. Step 6 ^ 7 (Scheme 1) Azaquinazoline formation Alternatively amino benzoic acid amide derivative of general formula 6 (which is commercially available or described in the literature) can be con verted to the corresponding azaquinazoline 7 in analogy to literature procedures. Typically derivative 6 is reacted with acetic acid at elevated temperature. For example see Bioorganic and Medicinal Chemistry Letters, 2008, vol. 18, # 3 p. 1037 – 1041 and references therein. Step 7 ^ 8 (Scheme 1) Conversion of hydroxyl group into leaving group In the next step (scheme 1) hydroxy azaquinazoline d erivative 7 can be converted to the corresponding azaquinazoline 8 in analogy to literatur e procedures. For W = chloro typically trichlorophosphate or thiony lchloride, with or without N,N‐dimethylaniline or N,N‐ diisopropylethylamine with or without an or ganic solvent such as for example toluene at elevated temperatures is used. For examples see Bioor ganic and Medicinal Chemistry Letters, 2011, 1270; Journal of Medicinal Chemistry, 2009, 2341; ACS Medicinal Chemistry Letters, 2013, 846; Bioorganic and Medicinal Chemistry Letters, 2010, 23 30; US6350750 or WO2015/54572 and references therein. For W = bromo typically phosphorus oxytribromide, wit h or without N,N‐dimethylaniline or N,N‐ diisopropylethylamine with or without an organic solve nt such as for example toluene at elevated temperatures is used. For examples see US2012/53174; WO2012/30912 or WO2012/66122 and references therein. For W = 2,4,6‐triisopropylsulfonate typically 2,4,6 triisopropylbenzenesulfonyl chloride, a base such as for example triethylamine and/or DMAP in an organ ic solvent such as for example dichloromethane is used. For examples see WO2010/99379 US2012/53176 and references therein. For W = tosylate typically 4‐methylbenzene‐1‐sulf onyl chloride, a base such as for example triethylamine or potassium carbonate and/or DMAP in a n organic solvent such as for example dichloromethane or acetonitrile is used. For examples see Organic Letters, 2011, 4374 or Bioorganic and Medicinal Chemistry Letters, 2013, 2663 and refer ences therein. For W = trifluoromethanesulfonate typically N,N‐bis(t rifluoromethylsulfonyl)aniline or trifluoromethanesulfonic anhydride, a base such as for example triethylamine or 1,8‐ diazabicyclo[5.4.0]undec‐7‐ene and/or DMAP in an or ganic solvent such as for example dichloromethane is used. For examples see Journal of the American Chemical Society, 2015, 13433 or WO2014/100501and references therein. Scheme 1. Synthesis route for the preparation of compounds of general formula (I), which are compounds of general formula (I), in which R2, A and x has the meaning as given for general formula (I), supra. Step 9 ^ 10 (Scheme 1) Acetyl formation In the first step (scheme 1) the bromo derivative 9 (which is commercially available or described in the literature) could be converted to the corresponding acetyl 10 in analogy to the numerous literature procedures. For example the reaction can be performed using different chemistries known to those skilled in the art, for example, Grignard chemistry using magnesium in an organic solvent as for example THF; or palladium catalyzed chemistry or Stille chemistry. For such transformations see the teachings of (Grignard: Fillon et al., Tetahedron 2003, 59, 8199; Leazer et al., Org. Synth.2005, 82, 115; Palladium: WO2005/5382; Stille: WO2019/122129 and the references therein. Step 10 ^ 11 (Scheme 1) Sulfinimine formation In the first step (scheme 1) aldehyde derivative 10 (which is commercially available or described in the literature) could be converted to the corresponding sulfinimine 11 in analogy
to the numerous literature procedures. For example the reaction could be performed at ambient temperature using Titanium(IV)ethoxide or Titanium(IV) isopropoxide in an organic solvent as for example THF. For a review about sulfinimine chemistry see for example Chem. Rev.2010, 110, 3600–3740; Chem. Soc. Rev.2009, 38, 1162–1186; Tetrahedron 2004, 60, 8003 or WO2019/122129 and the references therein. Step 11 ^ 12 (Scheme 1) Formation of sulfinamide In the next step (scheme 1) sulfinimine 11 can be converted to the corresponding sulfinamide 12 in analogy to the numerous literature procedures. For example the reaction can be performed using a reducing agent, for example, sodium borohydride or borane-THF, in a protic organic solvent as for example ethanol or methanol or tetrahydrofuran. Such transformations are known to those skilled in the art, see the teachings of Pan et al., Tetrahedron Asym., 2011, 22, 329; WO2019/122129; Li et al., Chem. Med. Chem., 2018, 13, 1363; Ghosh et al., Eur. J. Med. Chem., 2018, 160, 171. Alternatively, the reaction can be performed using a reducing agent, for example, diisopropylaluminium hydride, in an aprotic solvent, for example, toluene. Such transformations are known to those skilled in the art, see the teachings of WO2017/6282; Lee et al., Synlett., 2019, 30, 401. Step 12 ^ 13 (Scheme 1) Formation of amine In the next step (scheme 2) sulfinamide 12 can be converted to the corresponding amine 13 in analogy to the numerous literature procedures. For example the reaction can be performed using acetylchloride in a protic organic solvent as for example methanol. For a review about sulfinimine and sulfonamide chemistry see for example Chem. Rev.2010, 110, 3600–3740; Chem. Soc. Rev.2009, 38, 1162–1186; Tetrahedron 2004, 60, 8003 or WO2013030138 and the references therein.
Scheme 2 Synthesis route for the preparation of compounds of general formula (I), which are compounds of general formula (I), in which R2, A and x has the meaning as given for general formula (I), supra. Step 10 ^ 14 (Scheme 2) Formation of alcohol In the first step (scheme 2) ketone derivative 10 (which is commercially available or described in the literature) could be converted to the corresponding chiral alcohol 14 in analogy to the numerous literature procedures. For example the enanioselective reduction could be performed using catalytic hydrogenation, with hydrogen gas under pressure with a catalyst, for example a BINAP-derived catalyst, e.g. (R)- or (S)-RUCY-Xyl-BINAP (see WO2019/122129 page 140 or WO2013/185103 page 81). Step 14 ^ 15 (Scheme 2) Formation of azide In the next step (scheme 2) alcohol 14 can be converted to the corresponding azide 15 in analogy to the numerous literature procedures. For example the reaction can be performed using diphenylphosphonic azide and a base, for example, DBU, in an aprotic organic solvent as for example, toluene (see the teachings of WO2019/122129 page 144). For a review about azide chemistry see for example Chem. Rev.1988, 88, 297. Step 15 ^ 13 (Scheme 2) Formation of amine In the next step (scheme 2) azide 15 can be converted to the corresponding amine 13 in analogy to the numerous literature procedures. For example the reaction can be performed using the Staudinger reduction conditions, with a phosphine, for example, triphenyl phosphine, in water with various different organic solvents, for example methanol, ethanol or THF. Alternatively, the azide reduction can be carried out using catalytic hydrogenation methods, using a metal catalyst, for example, palladium on charcoal, under a pressurized atmosphere of hydrogen (see WO2019/122129 page 144). For a review about azide chemistry see for example Chem. Rev.1988, 88, 297. Scheme 3. Synthesis route for the preparation of compounds of general formula (I), which are compounds of general formula (I), in which R2, A and x has the meaning as given for general formula (I), supra. To those skilled in the art it is possible to carry out the chemical reactions described in Schemes 1 and 2, where the stereoisomers can be separated using various methods known to those skilled in the art, such as, for example, separation using chiral HPLC purification. The separation of these stereoisomers can be carried out on compounds of general formula 13. Scheme 4: Route for the preparation of compounds of general formula 16 (a compound of general formula I), wherein T, V, R 1 , R 2 , x, y and A have the meaning as given for general formula (I), supra and LG has the meaning as a leaving group, preferab ly chloro, bromo or a sulfonate group as depicted in scheme 4. Specific examples are described in the subsequent paragraphs. Step 12 + 8 ^ 17 (Scheme 4)
Amine coupling In the first step (scheme 4) amine derivative rac‐ 13 and azaquinazoline derivative 8 are converted to amine 16 in analogy to literature procedures. Typical ly the reaction is performed in an organic solvent such as for example THF, DMF, acetonitrile d ichloromethane or isopropyl alcohol with or without a base such as for example triethylamine, N ethyl‐N,N‐diisopropylamine, potassium carbonate or potassium tert‐butylate. For LG = chloro see for example the literature refe rences WO2008/86462; WO2008/86462 or European Journal of Medicinal Chemistry, 2015, 462 an d references therein. For LG = bromo see for example the literature refer ences US2009/247519 or Journal of Organic Chemistry, 2009, 8460 and references therein. For LG = tosylate see for example the literature re ferences Synthetic Communications, 2012, 1715; Synthesis 2015, 2055 or Bioorganic and Medicinal Chem istry Letters, 2013, 2663 and references therein. For LG = triflate see for example the literature re ferences Bioorganic and Medicinal Chemistry Letters, 2013, 3325 and references therein. For LG = 2,4,6‐triisopropylbenzenesulfonate see for example the literature reference WO2010/99379 and references therein. In accordance with a further aspect, the present inv ention covers intermediate compounds which are useful in the preparation of compounds of the presen t invention of general formula (I), particularly in the methods described herein. The present invention covers the intermediate compounds which are disclosed in the Example Section of this text, infra. The present invention covers any sub‐combination wit hin any embodiment or aspect of the present invention of intermediate compounds. In accordance with another aspect, the present invent ion covers methods of preparing compounds of the present invention, said methods comprising the step as described below and / or the Experimental Section. The preparation of compounds of general formula I ca n be performed in a protic or aprotic solvent, preferably in dioxan, tetrahydrofuran, N,N‐dimethylfor mamide, dimethylsulfoxid, methanol, ethanol or 2‐propanol.
Preferred bases which can be used for the prepara tion of compounds of the general formula I are N,N‐diisopropylethylamin or triethylamin. Said compound of general formula I can then optionally be converted into solvates, salts and/or solvates of such salts using the corresponding (i) s olvents and/or (ii) bases or acids. The present invention covers methods of preparing compounds of th e present invention of general formula (I), said methods comprising the steps as de scribed in the Experimental Section herein. The compounds of general formula (I) of the present invention can be converted to any salt, preferably pharmaceutically acceptable salts, as descri bed herein, by any method which is known to the person skilled in the art. Similarly, any salt of a compound of g eneral formula (I) of the present invention can be converted into the free compound, by any method which is known to the person skilled in the art. One of the most fundamental characteristics of cancer cells is their ability to sustain chronic proliferation whereas in normal tissues the entry int o and progression through the cell division cycle is tightly controlled to ensure a homeostasis of cell number and maintenance of normal tissue function. Loss of proliferation control is emphasized as one of the six hallmarks of cancer [Hanahan D and Weinberg 15 RA, Cell 100, 57, 2000; Hanahan D and Weinberg RA, Cell 144, 646, 2011]. Compounds of general formula (I) of the present inve ntion demonstrate a valuable pharmacological spectrum of action which could not have been predict ed. Compounds of the present invention have surprisingly been found to effectively inhibit the Ra s‐Sos1 interaction and it is possible therefore tha t said compounds be used for the treatment or prophyla xis of diseases, preferably hyperproliferative disorders in humans and animals. Compounds of the present invention can be utilized to inhibit, block, reduce, decrease, etc., cell proliferation and/or cell division, and/or produce apo ptosis. This method comprises administering to a mammal in need thereof, including a human, an amount of a compound of gener al formula (I) of the present invention, or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate, solvate or ester thereof, which is effective to treat the disorder. Hyperproliferative disorders include, but are not limi ted to, for example: psoriasis, keloids, and other hyperplasias affecting the skin, benign prostate hyper plasia (BPH), solid tumours, such as cancers of the breast, respiratory tract, brain, reproductive org ans, digestive tract, urinary tract, eye, liver, skin , head and neck, thyroid, parathyroid and their distant metastases. Those disorders also include lymphomas, sarcomas, and leukaemias. Examples of breast cancers include, but are not limi ted to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carc inoma in situ.
Examples of cancers of the respiratory tract inclu de, but are not limited to, small‐cell and non‐s mall‐ cell lung carcinoma, as well as bronchial adenoma an d pleuropulmonary blastoma. Examples of brain cancers include, but are not limited to, brain stem and hypophtalmic glioma, cerebellar and cerebral astrocytoma, medulloblastoma, ependymoma, as w ell as neuroectodermal and pineal tumour. Tumours of the male reproductive organs include, but are not limited to, prostate and testicular cancer. Tumours of the female reproductive organs include, but are not limited to, endometrial, cervical, ovarian, vaginal, and vulvar cancer, as well as sarc oma of the uterus. Tumours of the digestive tract include, but are not limited to, anal, colon, colorectal, oesophageal, gallbladder, gastric, pancreatic, rectal, small‐intest ine, and salivary gland cancers. Tumours of the urinary tract include, but are not limited to, bladder, penile, kidney, renal pelvis, ureter, urethral and human papillary renal cancers. Eye cancers include, but are not limited to, intraoc ular melanoma and retinoblastoma. Examples of liver cancers include, but are not limited to, hepatocellular carcinoma (liver cell carcinomas with or without fibrolamellar variant), cholangiocarcinoma (intrahepatic bile duct carcinoma), and mixed hepatocellular cholangiocarcinoma. Skin cancers include, but are not limited to, squamous cell carcinoma, Kaposi’s sarcom a, malignant melanoma, Merkel cell skin cancer, and non‐melanoma skin cancer. Head‐and‐neck cancers include, but are not limited to, laryngeal, hypopharyngeal, nasopharyngeal, oropharyngeal cancer, lip and oral cavity cancer and squamous cell. Lymphomas include, but are not limited to, AIDS‐related lymphoma, non‐Hodgkin’s lymphoma, cutaneous T‐cell lymphoma, Burkitt lymphoma, Hodgkin’s disease, and lymphoma of the central nervous system. Sarcomas include, but are not limited to, sarcoma of the soft tissue, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, and rhabdomyosarcoma. Leukemias include, but are not limited to, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leuke mia, and hairy cell leukemia. The present invention also provides methods of treating angiogenic disorders including diseases associated with excessive and/or abnormal angiogenesis.
Inappropriate and ectopic expression of angiogenesis can be deleterious to an organism. A number of pathological conditions are associated with the gr owth of extraneous blood vessels. These include, for example, diabetic retinopathy, ischemic retinal‐vein occlusion, and retinopathy of prematurity [Aiello et al., New Engl. J. Med., 1994, 331, 1480 ; Peer et al., Lab. Invest., 1995, 72, 638], age related macular degeneration (AMD) [Lopez et al., Invest. Opt hthalmol. Vis. Sci., 1996, 37, 855], neovascular glaucoma, psoriasis, retrolental fibroplasias, angiofibroma, inflammation, rheumatoid arthritis (RA), restenosis, in‐stent restenosis, vascular graft reste nosis, etc. In addition, the increased blood supply associated with cancerous and neoplastic tissue, encourages growth, leading to rapid tumour enlargement and metastasis. Moreover, the growth of new blood and lymph vessels in a tumour provides an escape route for renegade cells, encourag ing metastasis and the consequence spread of the cancer. Thus, compounds of general formula (I) o f the present invention can be utilized to treat and/or prevent any of the aforementioned angiogenesis disorders, for example by inhibiting and/or reducing blood vessel formation; by inhibiting, blocking, reducing, decreasing, etc. endothelial cell proliferation, or other types involved in angiogenesis, as well as causing cell death or a poptosis of such cell types. These disorders have been well characterized in humans, but also exist with a similar etiology in other mammals, and can be treated by administering pharmaceutical compositions of the present invention. The term “treating” or “treatment” as stated throughout this document is used conventionally, for example the management or care of a subject for the purpose of combating, alleviating, reducing, relieving, improving the condition of a disease or d isorder, such as a carcinoma. The compounds of the present invention can be used in particular in therapy and prevention, i.e. prophylaxis, of tumour growth and metastases, especially in solid tumours of all indications and stages with or without pre‐treatment of the tumour growth. Generally, the use of chemotherapeutic agents and/or anti‐cancer agents in combination with a compound or pharmaceutical composition of the present invention will serve to: 1. yield better efficacy in reducing the growth of a tumour or even eliminate the tumour as compared to administration of either agent alone, 2. provide for the administration of lesser amounts of the administered chemotherapeutic agents, 3. provide for a chemotherapeutic treatment that is well tolerated in the patient with fewer deleterious pharmacological complications than observed with single agent chemotherapies and certain other combined therapies,
4. provide for treating a broader spectrum of different cancer types in mammals, especially humans, 5. provide for a higher response rate among treated pat ients, 6. provide for a longer survival time among treated patients compared to standard chemotherapy treatments, 7. provide a longer time for tumour progression, and/or 8. yield efficacy and tolerability results at least as good as those of the agents used alone, compared to known instances where other cancer agent combinations produce antagonistic effects. In addition, the compounds of general formula (I) of the present invention can also be used in combination with radiotherapy and/or surgical intervent ion. In a further embodiment of the present invention, the compounds of general formula (I) of the present invention may be used to sensitize a cell to radiation, i.e. treatment of a cell with a compound of the present invention prior to radiation treatment of the cell renders the cell more susceptible to DNA damage and cell death than the c ell would be in the absence of any treatment with a compound of the present invention. In one aspect, the cell is treated with at least one compound of general formula (I) of the present inven tion. Thus, the present invention also provides a method o f killing a cell, wherein a cell is administered on e or more compounds of the present invention in combin ation with conventional radiation therapy. The present invention also provides a method of rendering a cell more susceptible to cell death, wherein the cell is treated with one or more compounds of general formula (I) of the present invention prior to the treatment of the cell to cau se or induce cell death. In one aspect, after the cell is treated with one or more compounds of general formula (I) of the present invention, the cell is treated with at least one compound, or at least one method, or a combination thereof, in order to cause DNA damage for the purpose of inhibiting the function of the normal cell or killing the cell. In other embodiments of the present invention, a cell is killed by treating the cell with at least one DNA damaging agent, i.e. after treating a cell with one or more compounds of general formula (I) of the present invention to sensitize the cell to cell death, the cell is treated with at least one DNA damaging agent to kill the cell. DNA damaging agents useful in the present invention include, but are not limited to, chemotherapeutic agents (e.g. cis platin), ionizing radiation (X‐rays, ultraviolet radiation), carcinogenic agents, and mutagenic agents.
In other embodiments, a cell is killed by treatin g the cell with at least one method to cause or i nduce DNA damage. Such methods include, but are not limited to, activation of a cell signalling pathway that results in DNA damage when the pathway is acti vated, inhibiting of a cell signalling pathway that results in DNA damage when the pathway is inhibited, and inducing a biochemical change in a cell, wherein the change results in DNA damage. By way of a non‐limiting example, a DNA repair pathway in a cell can be inhibited, thereby preventing the repair of DNA damage and resulting in an abnormal accumulation of DNA damage in a cell. In one aspect of the invention, a compound of general formula (I) of the present invention is administered to a cell prior to the radiation or ot her induction of DNA damage in the cell. In another aspect of the invention, a compound of general formu la (I) of the present invention is administered to a cell concomitantly with the radiation or other induction of DNA damage in the cell. In yet another aspect of the invention, a compound of general formula (I) of the present invention is administered to a cell immediately after radiation or other induction of DNA damage in the cell has begun. In another aspect, the cell is in vitro. In another embodiment, the cell is in vivo. It is possible for the compounds according to the i nvention to have systemic and/or local activity. For this purpose, they can be administered in a suitable manner, such as, for example, via the oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, vaginal, dermal, transdermal, conjunctival, otic route or as an implant or stent. For these administration routes, it is possible for the compounds according to the invention to be administered in suitable administration forms. For oral administration, it is possible to formulate the compounds according to the invention to dosage forms known in the art that deliver the compounds of the invention rapidly and/or in a modified manner, such as, for example, tablets (uncoa ted or coated tablets, for example with enteric or controlled release coatings that dissolve with a delay or are insoluble), orally‐disintegrating tablets, films/wafers, films/lyophylisates, capsules (for example hard or soft gelatine capsules), sugar‐coated tablets, granules, pellets, powders, emu lsions, suspensions, aerosols or solutions. It is possible to incorporate the compounds according to the invention in crystalline and/or amorphised and/or dissolved form into said dosage forms. Parenteral administration can be effected with avoidance of an absorption step (for example intravenous, intraarterial, intracardial, intraspinal, intralumbal or intratumoral) or with inclusion of absorption (for example intramuscular, subcutaneous, intracutaneous, percutaneous or
intraperitoneal). Administration forms which are sui table for parenteral administration are, inter alia, preparations for injection and infusion in the form of solutions, suspensions, emulsions, lyophylisates or sterile powders. Examples which are suitable for other administration routes are pharmaceutical forms for inhalation [inter alia powder inhalers, nebulizers], nasal drops, nasal solutions, nasal sprays; tablets/films/wafers/capsules for lingual, sublingual or buccal administration; suppositories; eye drops, eye ointments, eye baths, ocular inserts, ear drops, ear sprays, ear powders, ear‐ri nses, ear tampons; vaginal capsules, aqueous suspensions (lotions, mixturae agitandae), lipophilic suspensions, emulsions, ointments, creams, transdermal therapeutic systems (such as, for example, patches), milk, pastes, foams, dusting powders, implan ts or stents. The compounds according to the invention can be inco rporated into the stated administration forms. This can be effected in a manner known per se by mixing with pharmaceutically suitable excipients. Pharmaceutically suitable excipients include, inter ali a, ^ fillers and carriers (for example cellulose, microcrystalline cellulose (such as, for example, Avicel ® ), lactose, mannitol, starch, calcium phosphate (such as, for example, Di‐Cafos ® )), ^ ointment bases (for example petroleum jelly, paraffins , triglycerides, waxes, wool wax, wool wax alcohols, lanolin, hydrophilic ointment, polyethyle ne glycols), ^ bases for suppositories (for example polyethylene glyc ols, cacao butter, hard fat), ^ solvents (for example water, ethanol, isopropanol, gly cerol, propylene glycol, medium chain‐ length triglycerides fatty oils, liquid polyethylene g lycols, paraffins), ^ surfactants, emulsifiers, dispersants or wetters (for example sodium dodecyl sulfate), lecithin, phospholipids, fatty alcohols (such as, for example, Lanette ® ), sorbitan fatty acid esters (such as, for example, Span ® ), polyoxyethylene sorbitan fatty acid esters (s uch as, for example, Tween ® ), polyoxyethylene fatty acid glycerides (such a s, for example, Cremophor ® ), polyoxethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, glycerol fatty acid esters, poloxamers (such as, for example, Pluronic ® ), ^ buffers, acids and bases (for example phosphates, carbonates, citric acid, acetic acid, hydrochloric acid, sodium hydroxide solution, ammonium carbonate, trometamol, triethanolamine), ^ isotonicity agents (for example glucose, sodium chlori de), ^ adsorbents (for example highly‐disperse silicas),
^ viscosity‐increasing agents, gel formers, thickeners and/or binders (for example polyvinylpyrrolidone, methylcellulose, hydroxypropylmethylcellulose, hydroxypropyl‐ cellulose, carboxymethylcellulose‐sodium, starch, carbomers, polyacrylic acids (such as, for example, Carbopol ® ); alginates, gelatine), ^ disintegrants (for example modified starch, carboxymethylcellulose‐sodium, sodium starch glycolate (such as, for example, Explotab ® ), cross‐ linked polyvinylpyrrolidone, croscarmellose‐sodium (such as, for example, AcDiSol ® )), ^ flow regulators, lubricants, glidants and mould release agents (for example magnesium stearate, stearic acid, talc, highly‐disperse silicas (such as, for example, Aerosil ® )), ^ coating materials (for example sugar, shellac) and film formers for films or diffusion membranes which dissolve rapidly or in a modified manner (for example polyvinylpyrrolidones (such as, for example, Kollidon ® ), polyvinyl alcohol, hydroxypropylmethylcellulose, hydroxypropylcellulose, ethylcellulose, hydroxypropyl‐ methylcellulose phthalate, cellulose acetate, cellulose acetate phthalate, polyacrylates, polymethacrylates such as, for example, Eudragit ® )), ^ capsule materials (for example gelatine, hydroxypropylm ethylcellulose), ^ synthetic polymers (for example polylactides, polyglycolides, polyacrylates, polymethacrylates (such as, for example, Eudragit ® ), polyvinylpyrrolidones (such as, for example, Kollidon ® ), polyvinyl alcohols, polyvinyl acetates, polyet hylene oxides, polyethylene glycols and their copolymers and blockcopolymers), ^ plasticizers (for example polyethylene glycols, propyle ne glycol, glycerol, triacetine, triacetyl citrate, dibutyl phthalate), ^ penetration enhancers, ^ stabilisers (for example antioxidants such as, for ex ample, ascorbic acid, ascorbyl palmitate, sodium ascorbate, butylhydroxyanisole, butylhydroxytoluen e, propyl gallate), ^ preservatives (for example parabens, sorbic acid, thiomersal, benzalkonium chloride, chlorhexidine acetate, sodium benzoate), ^ colourants (for example inorganic pigments such as, for example, iron oxides, titanium dioxide), ^ flavourings, sweeteners, flavour‐ and/or odour‐maski ng agents.
The present invention furthermore relates to a pha rmaceutical composition which comprise at least one compound according to the invention, conventionally together with one or more pharmaceutically suitable excipient(s), and to their u se according to the present invention. In accordance with another aspect, the present invention covers pharmaceutical combinations, in particular medicaments, comprising at least one compound of general formula (I) of the present invention and at least one or more further active i ngredients, in particular for the treatment and/or prophylaxis of a hyper‐proliferative disorder, in pa rticular cancer. Particularly, the present invention covers a pharmaceu tical combination, which comprises: ^ one or more first active ingredients, in particular compounds of general formula (I) as defined supra, and ^ one or more further active ingredients, in particular those used for treatment of hyper‐ proliferative disorder, in particular cancer. The term “combination” in the present invention is used as known to persons skilled in the art, it being possible for said combination to be a fixed c ombination, a non‐fixed combination or a kit‐of‐ parts. A “fixed combination” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein, for example, a first active ingredient, such as one or more compounds of general formula (I) of the present invention, and a further active ingredient are present together in one unit dosage or in one singl e entity. One example of a “fixed combination” i s a pharmaceutical composition wherein a first active ingredient and a further active ingredient are present in admixture for simultaneous administration, such as in a formulation. Another example of a “fixed combination” is a pharmaceutical combination wherein a first activ e ingredient and a further active ingredient are present in one unit without be ing in admixture. A non‐fixed combination or “kit‐of‐parts” in the present invention is used as known to persons skilled in the art and is defined as a combination wherein a first active ingredient and a further active ingredient are present in more than one unit. One e xample of a non‐fixed combination or kit‐of‐part s is a combination wherein the first active ingredient and the further active ingredient are present separately. It is possible for the components of the non‐fixed combination or kit‐of‐parts to be administered separately, sequentially, simultaneously, c oncurrently or chronologically staggered. The compounds of the present invention can be admini stered as the sole pharmaceutical agent or in combination with one or more other pharmaceutically active ingredients where the combination causes no unacceptable adverse effects. The present invention also covers such pharmaceutical
combinations. For example, the compounds of the pr esent invention can be combined with known anti‐tumor agents (cancer therapeutics). Examples of anti‐tumor agents (cancer therapeutics) include: 131I‐chTNT, abarelix, abiraterone, aclarubicin, ado‐trastuzumab emtansine, afatinib, aflibercept, aldesleukin, alectinib, alemtuzumab, alendronic acid, alitretinoin, altretamine, amifostine, aminoglutethimide, hexyl aminolevulinate, amrubicin, amsacrine, anastrozole, ancestim, anethole dithiolethione, anetumab ravtansine, angiotensin II, antithrombin III, aprepitant, arcitumomab, arglabin, arsenic trioxide, asparaginase, axitinib, azacitidine, basilixim ab, belotecan, bendamustine, besilesomab, belinostat, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, blinatumomab, bortezomib, buserelin, bosutinib, brentuximab vedotin, busulfan, cabazitaxel, cabozantinib, calcitonine, calcium folinate, calcium levofolinate, capecitabine, capromab, carboplatin, carboquone, carfilzomib, carmofur, carmusti ne, catumaxomab, celecoxib, celmoleukin, ceritinib, cetuximab, chlorambucil, chlormadinone, chlormethine, cidofovir, cinacalcet, cisplatin, cladribine, clodronic acid, clofarabine, cobimetinib, copanlisib, crisantaspase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daratumumab, darbepoetin alfa, darolutamide, dabrafenib, dasatinib, daunorubicin, decitabine, degarelix, denileukin diftitox, denosumab, depreotide, deslorelin, dianhydrogalactitol, dexrazoxane, dibrospidium chloride, dianhydrogalactitol, diclofenac, dinutuximab, docetaxel, dolasetron, doxifluridine, doxorubicin, doxorubicin + estrone, dronabinol, eculizumab, edrecolomab, elliptinium acetate, elotuzumab, eltrombopag, endostatin, enocitabine, enzalutamide, epir ubicin, epitiostanol, epoetin alfa, epoetin beta, epoetin zeta, eptaplatin, eribulin, erlotinib, esomeprazole, estradiol, estramustine, ethinylestradiol, etoposide, everolimus, exemestane, fadrozole, fentanyl, filgrastim, fluoxymesterone, floxuridine, fludarabine, fluorouracil, flutamide, folinic acid, formestane, fosaprepitant, fotemustine, fulvestrant, gadobutrol, gadoteridol, gadoteric acid meglumine, gadoversetamide, gadoxetic acid, gallium nitrate, ganirelix, gefitinib, gemcitabine, gemtuzumab, Glucarpidase, glutoxim, GM‐CSF, goserelin, granisetron, granulocyte colony stimulating factor, histamine dihydrochloride, histrelin, hydroxycarbamide, I‐125 seeds, lansoprazole, ibandronic acid, ibritumomab tiuxetan, ibrutinib, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, indisetron, incadronic acid, ingenol mebutate, interferon alfa, interferon beta, interferon gamma, iobitridol, iobenguane (123I), iomeprol, ipilimumab, irinotecan, It raconazole, ixabepilone, ixazomib, lanreotide, lansoprazole, lapatinib, larotrectinib, Iasocholine, lenalidomide, lenvatinib, lenograstim, lentinan, letrozole, leuprorelin, levamisole, levonorgestrel, levothyroxine sodium, lisuride, lobaplatin, lomustine, lonidamine, masoprocol, medroxyprogesterone, megestrol, melarsoprol, melphalan, mepitiostane, mercaptopurine, mesna, methadone, methotrexate, methoxsalen,
methylaminolevulinate, methylprednisolone, methyltestosterone, metirosine, mifamurtide, miltefosine, miriplatin, mitobronitol, mitoguazone, mito lactol, mitomycin, mitotane, mitoxantrone, mogamulizumab, molgramostim, mopidamol, morphine hydroch loride, morphine sulfate, nabilone, nabiximols, nafarelin, naloxone + pentazocine, naltrexo ne, nartograstim, necitumumab, nedaplatin, nelarabine, neridronic acid, netupitant/palonosetron, ni volumabpentetreotide, nilotinib, nilutamide, nimorazole, nimotuzumab, nimustine, nintedanib, nitracri ne, nivolumab, obinutuzumab, octreotide, ofatumumab, olaparib, omacetaxine mepesuccinate, omepraz ole, ondansetron, oprelvekin, orgotein, orilotimod, osimertinib, oxaliplatin, oxycodone, oxymetholone, ozogamicine, p53 gene therapy, paclitaxel, palbociclib, palifermin, palladium‐103 see d, palonosetron, pamidronic acid, panitumumab, panobinostat, pantoprazole, pazopanib, pegaspargase, PEG‐epoetin beta (methoxy PEG‐epoetin beta), pembrolizumab, pegfilgrastim, peginterferon alfa 2b, pemetrexed, pentazocine, pentostatin, peplomycin, Perflubutane, perfosfamide, Pertuzumab, pici banil, pilocarpine, pirarubicin, pixantrone, plerixafor, plicamycin, poliglusam, polyestradiol phosphate, polyvinylpyrrolidone + sodium hyaluronate, polysaccharide‐K, pomalidomide, ponatinib, porfimer sodium, pralatrexate, prednimustine, prednisone, procarbazine, procodazole, propranolol, quinagolide, rabeprazole, racotumomab, radium‐223 chloride, radotinib, raloxifene, raltitrexed, ramosetron, ramucirumab, ranimustine, rasburicase, razoxane, refametinib, regorafenib, risedronic acid, rhenium‐186 etidronate, rituximab, rogaratinib, rolapitant, romidepsin, romiplostim, romurtide, roniciclib, samarium (153Sm) lexidronam, sargramostim, satumomab, secretin, siltuximab, sipuleucel‐T, sizofiran, sobuzoxane, sodium glycididazole, sonidegib, sorafenib, stanozolol, streptozocin, sunitinib, talaporfin, talimogene laherparepvec, tamibarotene, tamo xifen, tapentadol, tasonermin, teceleukin, technetium (99mTc) nofetumomab merpentan, 99mTc‐HYNIC‐[Tyr3]‐octreotide, tegafur, tegafur + gimeracil + oteracil, temoporfin, temozolomide, temsiro limus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thyrotropin alfa, t ioguanine, tocilizumab, topotecan, toremifene, tositumomab, trabectedin, trametinib, tramadol, trastuzumab, trastuzumab emtansine, treosulfan, tretinoin, trifluridine + tipiracil, trilostane, triptorelin, trametinib, trofosfamide, thrombopoietin, tryptophan, ubenimex, valatinib, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib, vorinostat, vorozole, yttrium‐90 glass microspheres, zinostatin, zinostatin stimalamer, zoledro nic acid, zorubicin. Further examples of combination partners are ATR inhibitors (e.g. BAY 1895344), DHODH inhibitors (e.g. BAY 2402234), SHP2 inhibitors (e.g. SHP099, RMC‐4550, TNO155) or H‐, N‐ or K‐Ras inhibitors, including inhibitors of mutants thereof, especially K‐RAS‐G12C inhibitors (e.g. ARS‐853, ARS‐1620, AMG‐510, MRTX849, MRTX1257) or farnesyl transferase inhibitors.
In particular, the present invention covers a comb ination of a covalent inhibitor of KRAS‐G12C and a SOS1 inhibitor. It has been shown that covalent KRAS‐G12C inhibitors (e.g. ARS‐853 or ARS‐1620) specifically bind to KRAS‐G12C in the GDP‐bound state, but not in the GTP‐bound state (Patricelli 2016 Cancer Discovery; Janes et al. 2018 Cell), ther eby trapping KRAS‐G12C in its inactive GDP‐bound state. In addition, it has been shown that certain RAS mutants, which usually exist in the active, GTP bound state, are undergoing a slow intrinsic GTP hyd rolysis, in particular G12C and G12D mutants of KRAS (Hunter et al. 2015 Molecular Cancer Research). It can be postulated that even those mutant RAS proteins require the activation by nucleotide exchange factors like SOS1 for full activity and tumorigenesis. Treatment with a SOS1 inhibitor is expected to shift the intracellular equilibrium of KRAS mutants towards the inactive GDP‐bound state, which in turn favours binding of inhibitors of KRAS which bind preferentially to the GDP‐bound sta te of RAS, as is the case for covalent KRAS‐G12C inhibitors like ARS‐853 and ARS‐1620. Synergistic anti‐proli ferative activity in vitro has been shown for the combination of BAY‐293 with ARS‐853 (Hill ig 2019 PNAS). Based upon standard laboratory techniques known to ev aluate compounds useful for the treatment of hyper‐proliferative disorders, by standard toxicity tests and by standard pharmacological assays for the determination of treatment of the conditions identified above in mammals, and by comparison of these results with the results of known active ingredients or medicaments that are used to treat these conditions, the effective dosage of the compounds of the present invention can readily be determined for treatment of each desired indication. The amount of the active ingredient to be administered in the treatment of one of these conditions can vary widely according to such considerations as the particular compound and dosage unit employed, the mode of administration, the period of treatment, the age and sex of the patient treated, and the nature and extent of the condition treated. The total amount of the active ingredient to be administered will generally range fr om about 0.001 mg/kg to about 200 mg/kg body weight per day, and preferably from about 0.01 mg/kg to about 20 mg/kg body weight per day. Clinically useful dosing schedules will range from one to three times a day dosing to once every four weeks dosing. In addi tion, it is possible for "drug holidays", in which a patient is not dosed with a drug for a certain period of time, to be beneficial to the overall balance between pharmacological effect and tolerability. It is possible for a unit dosage to contain from about 0.5 mg to about 1500 mg of active ingredient, and can be administered one or more times per day or less than once a day. The average daily dosage for administration by injection, including intravenous, intramuscular, subcutaneous and parenteral injections, and use of infusion techniques will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily rectal dosage regimen will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily vaginal dosage regimen
will preferably be from 0.01 to 200 mg/kg of total body weight. The average daily topical dosage regimen will preferably be from 0.1 to 200 mg administered between one to four times daily. The transdermal concentration will preferably be that requ ired to maintain a daily dose of from 0.01 to 200 mg/kg. The average daily inhalation dosage regime n will preferably be from 0.01 to 100 mg/kg of total body weight. Of course the specific initial and continuing dosage regimen for each patien t will vary according to the nature and severity of the condition as determin ed by the attending diagnostician, the activity of the specific compound employed, the age and general condition of the patient, time of administration, route of administration, rate of excretion of the drug, drug combinations, and the like. The desired mode of treatment and number of d oses of a compound of the present invention or a pharmaceutically acceptable salt or ester or composition thereof can be ascertained by those skilled in the art using conventional treatment tests . EXPERIMENTAL SECTION The following table lists the abbreviations used in this paragraph, and in the examples section. BuLi Butyllithium DCE Dichloroethane DCM Dichloromethane DMF Dimethylformamide DMSO Dimethyl sulfoxide EA Ethyl acetate FA Formic acid HPLC, LC high performance liquid chromatography h hour LiHMDS Lithium bis(trimethylsilyl)amide KHMDS Potassium bis(trimethylsilyl)amide KOtBu Potassium tert‐butoxide min minute LDA Lithiumdiisopropylamid MS mass spectroscopy
NMR nuclear magnetic resonance NaHMDS Sodium bis(trimethylsilyl)amide PE Petrol ether Rac Racemate R f Retardiation factor R t Retention time RT Room temperature TFA Trifluoroacetic acid THF Tetrahydrofuran TLC thin‐layer chromatography Chemical names were generated using ACD/Name Batch Ve rsion 12.01 or Autonom 2000. All reagents, for which the synthesis is not describ ed in the experimental part, are either commercially available or synthesized as described in literature references. Analytical Methods LC‐MS Method 1: Column: Ascentis Express C18 2.7 µm, 30x2.1 mm Fragment. potential: 50 V Mass range: 80‐800 m/z Solvent: A = H 2 O + 0.1%vol HCOOH B = methanol + 0.1%vol HCOOH Gradient: 0‐1 min 5% B, 1‐4 min 5‐100% B 4‐5 min 100 % B, 5‐6 min 100‐5% B, 6‐ 6.5 min 5% B Flow: 0.8 mL/min Temperature: 30°C Injection: 1.0 µL Detection: MM‐ES + APCI + DAD (254 nm) System time delay: 0.2 min LC‐MS Method 2: MS instrument type: Micromass Quatr o Micro; HPLC instrument type: Agilent 1100 Series; UV DAD; column: Chromolith Flash RP‐18E 25 2 mm; mobile phase A: 0.0375% TFA in water, mobile phase B: 0.01875% TFA in acetonitrile; gradien t: 0.0 min 100% A ^ 1.0 min 95% A ^ 3.0 min 95% A ^ 3.5 min 5% A ^ 3.51 min 5% A ^ 4.0 min 95% A; flow rate: 0.8 mL/min; column tem p: 50°C; UV detection: 220 nm & 254 nm. LC‐MS Method 3: System: Waters Acquity UPLC‐MS: Binary Solvent Manager, Samp le Manager/Organizer, PDA, ELSD
Column: Acquity UPLC BEH C18 1.7 µm, 50x2.1 mm Solvent: A = H 2 O + H 2 O + 0.1%vol. HCOOC (99%) B = acetonitrile Gradient: 0‐1.6 min 1‐99% B, 1.6‐2 min 99% B Flow: 0.8 mL/min Temperature: 60°C Injection: 2.0 µL Detection: DAD scan range 210‐400 nm + ELSD LC‐MS Method 4: System: Shimadzu LC‐MS: UFLC 20‐AD and LCMS 2020 MS dete ctor Column: Shim‐pack XR‐ODS 2.2 µm, 3.0x50 mm Solvent: A = H 2 O + 0.05%vol. HCOOC (99%) B = acetonitrile+ 0.05%vol. HCOOC (99%) LC‐MS Method 5: System: Waters Acquity UPLC‐MS: Binary Solvent Manager, Samp le Manager/Organizer, PDA, ELSD Column: Acquity UPLC BEH C18 1.7 µm, 50x2.1 mm Solvent: A = H 2 O + 0.2%vol. NH 3 (32%) B = acetonitrile Gradient: 0‐1.6 min 1‐99% B, 1.6‐2 min 99% B Flow: 0.8 mL/min Temperature: 60°C Injection: 2.0 µL Detection: DAD scan range 210‐400 nm + ELSD LC‐MS Method 6: System: Instrument HPLC: Waters UPLC Acquity; Instrument MS: Waters ZQ Column: Acquity UPLC BEH C18 1.7µm, 50x2.1mm Solvent: A = H 2 O + 0.1%vol. HCOOC (99%) B = acetonitrile Gradient: 0‐1.6 min 1‐99% B, 1.6‐1.8 min 99% B, 1.81‐2 min 1% B Flow: 0.8 mL/min Temperature: 60°C Detection: PDA scan range 210‐400 nm LC‐MS Method 7: System: Agilent 1290 UHPLC‐MS Tof Column: BEH C 18 (Waters) 1.7 µm, 50x2.1 mm Solvent: A = H 2 O + 0.05%vol. HCOOC (99%) B = acetonitrile + 0.05%vol. HCOOC (99%) Gradient: 0‐1.7 min 2‐90% B, 1.7‐2 min 90% B, 2‐2.5 m in 90‐2% B Flow: 1.2 mL/min Temperature: 60°C Detection: DAD scan range 210‐400 nm LC‐MS Method 8: System: Waters Acquity UPLC‐MS: Binary Solvent Manager, Samp le Manager/Organizer, PDA, ELSD
Column: Acquity UPLC BEH C18 1.7 µm, 50x2.1 mm Solvent: A = H 2 O + 0.1%vol. HCOOC (99%) B = acetonitrile Gradient: 0‐1.6 min 1‐99% B, 1.6‐2 min 99% B Flow: 0.8 mL/min Temperature: 60°C Injection: 2.0 µL Detection: DAD scan range 210‐400 nm + ELSD LC‐MS Method 9: System: Waters Acquity UPLC‐MS SingleQuad Column: Kinetex C 18 (Phenomenex) 2.6 µm, 50x2.1 mm Solvent: A = H 2 O + 0.05%vol. HCOOC (99%) B = acetonitrile + 0.05%vol. HCOOC (99%) Gradient: 0‐0.2 min 2% B, 0.2‐1.7 min 2‐90% B, 1.7‐1.9 min 90% B, 1.9‐2 min 90‐2% B, 2‐2.5 min 2% B Flow: 1.3 mL/min Temperature: 60°C Detection: DAD scan range 210‐400 nm LC‐MS method 10: System: Waters Acquity UPLC‐MS SingleQuad; Column: A cquity UPLC BEH C18 1.7 µm, 50x2.1mm; Solvent: A = H2O + 0.2%vol. NH3 (32%), B = acetoni trile; Gradient: 0‐1.6 min 1‐99% B, 1.6‐2 min 99% B; Flow: 0.8 mL/min; Temperature: 60°C; Detection: DAD scan range 210‐400 nm Preparative HPLC a) Autopurifier: acidic conditions System: Waters Autopurification system: Pump 2545, Sample Mana ger 2767, CFO, DAD 2996, ELSD 2424, SQD Column: XBrigde C18 5.0 µm 100x30 mm Solvent: A = H 2 O + 0.1%vol. HCOOH (99%) B = acetonitrile Gradient: 0‐0.5 min 5% B 25 mL/min, 0.51‐5.5 min 10‐100% B 70 mL/min, 5.51‐6.5 min 100% B 70 mL/min Temperature: RT Solution: max. 250 mg / max. 2.5 mL DMSO or DMF Injection: 1 x 2.5 mL Detection: DAD scan range 210–400 nm, MS ESI+, ESI‐, scan range 160‐1000 m/z b) Autopurifier: basic conditions System: Waters Autopurification system: Pump 2545, Sample Mana ger 2767, CFO, DAD 2996, ELSD 2424, SQD Column: XBrigde C18 5.0 µm 100x30 mm Solvent: A = H 2 O + 0.2%vol. NH 3 (32%) B = acetonitrile
Gradient: 0‐0.5 min 5% B 25 mL/min, 0.51‐5.5 min 10‐100% B 70 mL/min, 5.51‐6.5 min 100% B 70 mL/min Temperature: RT Solution: max. 250 mg / max. 2.5 mL DMSO or DMF Injection: 1 x 2.5 mL Detection: DAD scan range 210–400 nm, MS ESI+, ESI‐, scan range 160‐1000 m/z Method X1: Instrument: Labomatic HD5000, Labocord‐5000; Gilson G X‐241, Labcol Vario 4000; Column: Chiralpak IE 5 µm 250x20 mm; Eluent A: MTBE + 0.1%vol. Diet hylamine (99%); Eluent B: Ethanol; Isocratic: 90%A + 10%B; Flow 30.0 mL/min; UV 254 nm. Method X2: Instrument: Labomatic HD5000, Labocord‐5000; Gilson G X‐241, Labcol Vario 4000; Column: Chiralpak IA 5 µm 250x30 mm; Eluent A: MTBE + 0.1%vol. Diet hylamine (99%); Eluent B: Ethanol; Isocratic: 85%A + 15%B; Flow 40.0 mL/min; UV 254 nm. Method X3: Instrument: Labomatic HD5000, Labocord‐5000; Gilson G X‐241, Labcol Vario 4000, Column: Chiralpak IA 5.0 µm 250x30 mm; Eluent: 100% Acetonitrile; Flo w 50.0 mL/min; UV 280 nm. Method X4: Instrument: Waters Autopurification system; Column: Wat ers XBrigde C18 5.0 µm 100x30 mm; Eluent A: H 2 O + 0.2%vol. NH 3 (32%), Eluent B: Acetonitrile; Gradient: 0.00 0.50 min 8% B (25‐ >70mL/min), 0.51–5.50 min 8‐15% B (70mL/min), D AD scan: 210‐400 nm. Method X5: Instrument: Labomatic HD5000, Labocord‐5000; Gilson G X‐241, Labcol Vario 4000, Column: Chiralpak IF 5.0 µm 250x30 mm; Eluent A: Hexane + 0.1%vol. Diethylamine (99%); Eluent B: Ethanol; Isocratic: 90%A + 10%B; Flow 50.0 mL/min; UV 280 nm. Method X6: Instrument: Waters Autopurification system; Column: Wat ers XBrigde C18 5.0 µm 100x30 mm; Eluent A: H 2 O + 0.2%vol. NH 3 (32%), Eluent B: Acetonitrile; Gradient: 0.00 0.50 min 30% B (25‐ >70mL/min), 0.51–5.50 min 30‐45% B (70mL/min), DAD scan: 210‐400 nm. Method X7: Instrument: Labomatic HD5000, Labocord‐5000; Gilson G X‐241, Labcol Vario 4000, Column: Chiralpak ID 5.0 µm 250x30 mm; Eluent A: H exane + 0.1%vol Diethylamin (99%); Eluent B: 2‐ Propanol; Isocratic: 85%A + 15%B; Flow 50.0 mL/min; UV 254 nm.
Synthesis of intermediates 13 Experimental procedure [A] for the synthesis of 13‐ a (see WO 2019/122129, page 141, line 2 – page 144, line 1) A solution of 12‐a (13.20 g, 45.00 mmol; 1.0 equi v.) in 1,4‐dioxane (100 ml) is cooled to 0°C and treated with 4 N HCI in 1,4‐dioxane (50.00 ml, 20 0.00 mmol, 4.4 equiv.). The reaction mixture is stirred for 3 h. After complete conversion of the s tarting material, the reaction mixture is concentrated under reduced pressure, the precipitate f iltered and washed with diethyl ether to obtain the desired product 13‐a as HCI salt. The crude product 13 is purified by chromatography i f necessary and isolated as HCI salt. Experimental procedure [B] for the synthesis of B‐5 k (see WO 2019/122129, page 144, line 2 – page 146, line 1) Alcohol 14 (2.00 g, 9.61 m mol, 1.0 equiv.) is dissolved in anhydrous toluene ( 20 mL). Diazabicycloundecene (1.73 mL, 11.5 mmol, 1.2 equiv.) and diphenylphosphonic azide (2.28 mL, 10.6 mmol, 1.1 equiv.) are added subsequently. The reactio n mixture is stirred at 40°C for 18 h until complete conversion of 14 is achieved. The reaction mixture is cooled to room temperature and the organic layer is washed with aqueous Na 2 CO 3 solution (2 x 10 mL). Azide B‐7a thus obta ined is not isolated but directly converted in the next step. Pd/C (200 mg, 10% w/w, 10% Pd) is added to the or ganic layer. The reaction mixture is charged with a H 2 atmosphere (10 bar) and is stirred for 24 h until complete conversion of 15 is achieved. The reaction is filtered and the volatiles are removed i n vacuo. The residue is dissolved in methyl tert‐ butyl ether (30 mL) and treated with HCI in dioxane (4.8 mL, 4 M). The white precipitate is filter, washed with methyl tert‐butyl ether (20 mL) and fu rther dried in vacuo to furnish the desired product 13. The crude product is purified by chromat ography if necessary. Table 1: Intermediates 13 (benzyl amines) available i n analogous manner starting from different sulfonamides 12 (experimental procedure [A], table 1, column 2) or alcohols 14 via azides 15 (experimental procedure [B], table 1, column 3) Table 1:
The synthesis of the different necessary sulfonamides B‐4 is described in WO 2019/122129 at page 136 line 2 to page 140 line 9. The synthesis of the different necessary alcohols B 6 is described in WO 2019/122129 at page 140 line 10 to page 141 line 1 (incl. table 14). Intermediate 1 1‐bromo‐3‐(difluoromethyl)‐2‐fluorobenzene To a solution of 3‐bromo‐2‐fluorobenzaldehyde (4. 07 g, 20.1 mmol) in DCM (35 ml) at 0°C was added slowly dropwise a solution of N‐ethyl‐N‐(trifluor o‐lambda 4 ‐sulfanyl)ethanamine (4.0 ml, 30 mmol) in DCM (10ml). The reaction was allowed to warm and stirred at RT overnight. The reaction mixture was added to ice‐water and extracted with DCM. The organics were combined, washed with sat. NaCl(aq), filtered through an hydrophobic filter and concentrated under reduced pressure. The residue was purified by silica chromatography (Hexane: EtOAc) and gave the titled compound (3.57 g, 75%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 2.518 (0.97) , 2.522 (0.62), 7.113 (7.95), 7.248 (16.00), 7.303 (4.71), 7.323 (9.95), 7.343 (5.61), 7.383 (7.82), 7.6 42 (3.92), 7.659 (6.89), 7.678 (3.45), 7.911 (3.70), 7.928 (6.59), 7.948 (3.45). Intermediate 2
1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐1‐ one To a solution of 1‐bromo‐3‐(difluoromethyl)‐2‐ fluorobenzene (3.07 g, 13.6 mmol) in anhydrous THF (10ml) at ‐10°C was added isopropylmagnesium chlori de (2M in THF, 7.5 ml, 15 mmol). The reaction was stirred at ‐10°C for 1h and then to added t o acetic anhydride (3.9 ml, 41 mmol) cooled to ‐1 5°C. The reaction was was to warm to 0°C and stirred for 15 min. The reaction was quenched by the addition of water and stirred at 60°C for 15 min. The reaction mixture was extracted with DCM. The organics were combined, washed with sat. NaHCO3(aq), sat. NaCl(aq), filtered through an hydrophobic filter and concentrated under reduced pres sure. The crude product (787 mg, 28%) was used directly without any further purification. Intermediate 3 (R)‐N‐{1‐[3‐(difluoromethyl)‐2‐fluorophenyl]et hylidene}‐2‐methylpropane‐2‐sulfinamide To a solution of 1‐[3‐(difluoromethyl)‐2‐fluorophenyl]e than‐1‐one (787 mg, 4.18 mmol) and (R)‐2‐ methyl‐2‐propane‐2‐sulfinamide (760 mg, 6.27 mmol) was added Ti(OEt)4 (2.86 g, 12.5 mmol) and heated at 80°C overnight. The reaction was added to a mixture of EtOAc and ice‐water and extracted with EtOAc. The organics were combined, filtered through an hydrophobic filter and concentrated under reduced pressure. The residue (1.31 g, 97%) wa s used directly in the next step. Intermediate 4 (R)‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoroph enyl]ethyl}‐2‐methylpropane‐2‐sulfinamide To a solution of (R)‐N‐{1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethy lidene}‐2‐methylpropane‐2‐ sulfinamide (1218 mg, 4.18 mmol) in THF (12 ml) was cooled to 0°C and NaBH4 (158 mg, 4.18 mmol) was added. The reaction was stirred at RT for 2h. The reaction was added to a mixture of EtOAc and ice‐water, then extracted with EtOAc. The organics were combined, filtered through an hydrophobic
filter and concentrated under reduced pressure. The titled compound (802 mg, 62%) was obtained after silica chromatography (EtOAc:Hexane) along with its diastereoisomer (166 mg, 13%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.099 (16.00 ), 1.154 (0.44), 1.172 (0.85), 1.190 (0.42), 1.401 (2.11), 1.418 (2.10), 1.987 (1.59), 5.870 (0.54), 5.8 89 (0.52), 7.074 (0.41), 7.209 (0.86), 7.345 (1.03). Intermediate 5 (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan 1‐amine, salt with hydrogen chloride x HCl To an ice‐cooled solution of (R)‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}‐2‐ methylpropane‐2‐sulfinamide (1.00 g, 3.41 mmol) in dioxane (7.5 ml) was added HCl (4M in dioxane, 3.75 ml). The reaction was allowed to warm to RT and stirred for 3h. The reaction mixture was concentrated under reduced pressure to approximately a volume of about 2ml. The solid was collected by filtered and was washed with MTBE and the titled compound (618 mg, 76%) was obtained. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.102 (7.29), 1.532 (7.14), 1.549 (7.00), 2.518 (0.81), 2.523 (0.57), 3.072 (2.53), 3.565 (5.88), 4.636 (0.46), 4.6 53 (1.59), 4.670 (1.66), 4.681 (0.63), 4.686 (0.58), 5.760 (16.00), 7.119 (2.25), 7.254 (4.53), 7.388 (2.0 2), 7.429 (1.08), 7.449 (2.38), 7.468 (1.37), 7.651 (1.03), 7.669 (1.76), 7.687 (0.86), 7.888 (0.87), 7.9 06 (1.16), 7.925 (0.54), 8.584 (0.43), 8.709 (1.89). Intermediate 6 6‐Fluoro‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ol A round‐bottom flask was charged with 5.00 g (32.0 mmol, commercially available) 5‐Amino‐2‐ fluoro‐4‐pyridinecarboxylic acid, 7.57 g (80 mmol, commercially available) acetamidine hydrochloride , and 6.56 g (80 mmol) anhydrous sodium acetate. The mixture was suspended in 50.0 ml of 2‐ methoxyethanol, and then the mixture was stirred at 130 °C for 16 h. The course of the reaction was monitored by LC/MS. Complete conversion was observed. The resulting mixture was poured into cold water and stirred for 30 min. The precipitate was f iltered off and dried in vacuo. 5.95 g (98 % d. T h.) of the title compound was obtained in form of a be ige‐coloured solid. 1H‐NMR (400 MHz, DMSO‐d6): ^ [ppm] = 13.14‐11.96 (br s, 1H), 8.66 (s, 1H), 7.59 (d, 1H), 2.37 (s, 3H).
Intermediate 7 6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ol A round‐bottom flask was charged with ethanol (110 ml) and cooled with an ice bath. To the ethanol was carefully added sodium (3.73 g, 163 mmol) and s tirred for 5 min. 6‐fluoro‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐ol (5.85 g, 32.7 mmol) was added an d the mixture was stirred at 110 °C for 16 h. Th e course of the reaction was monitored by LC/MS, nearl y complete conversion was detected. The solution was cooled to room temperature and concentra ted in vacuo. Under cooling in an ice‐bath the residue was diluted with 500 ml of water, then acidified with 2M hydrochloric acid (200 mL) to pH = 1 and extracted with dichloromethane (2 x 200 ml) and a mixture of dichlormethane/isopropanol (4 : 1, 5 x 200 ml). The combined organic layers were dried over sodium sulfate and then concentrated in vacuo. The title co mpound (4.83 g, 77 %) was obtained in form of a beige/brown‐coloured solid. 1H‐NMR (400 MHz, DMSO): ^ [ppm] = 8.62 (s, 1H), 7.17 (s, 1H), 4.34 (q, 2H), 1.34 (t, 3H). Intermediate 8 6‐methoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ol A mixture of 5‐amino‐2‐methoxypyridine‐4‐carbox ylic acid (2.50 g, 14.9 mmol), ethanimidamide hydrochloride (2.81 g, 29.7 mmol) and anhydrous sodiu m acetate (2.44 g, 29.7 mmol) in 2‐ methoxyethanol (40 ml) was heated under reflux condit ions for 6h. The solution was cooled to room temperature and water (50 ml) was added. The precipi tate was collected by filtration, washed with water and dried in vacuo to give the titled compoun d (2.31 g). 1H‐NMR (400 MHz, DMSO): ^ [ppm] = 2.27 (br s, 1H ), 8.60 (d, 1H), 7.19 (d, 1H), 3.79‐3.98 (s, 3H), 2.32 (s, 3H). Intermediate 9 N‐[(3R)‐1‐(4‐hydroxy‐2‐methylpyrido[3,4‐d]pyri midin‐6‐yl)pyrrolidin‐3‐yl]acetamide
A mixture of 6‐fluoro‐2‐methylpyrido[3,4‐d]pyrimi din‐4‐ol (10.0 g, 55.8 mmol) and N‐[(3R)‐pyrro lidin‐ 3‐yl]acetamide (12.5 g, 97.7 mmol in DMSO (40 ml) was added triethylamine (23 ml, 170 mmol) and heated at 90°C for 16h. The reaction mixture was concentrated under reduced pressure and the residue purified by silica chromatography (DCM:EtOH) t o give the titled compound (13.56g, 80%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.035 (2.29) , 1.052 (4.89), 1.070 (2.52), 1.807 (16.00), 1.898 (0.76), 1.911 (0.63), 1.922 (0.47), 1.929 (0.47), 2.1 59 (0.51), 2.174 (0.60), 2.190 (0.55), 2.205 (0.43), 2.258 (0.80), 2.284 (13.66), 2.522 (1.32), 2.539 (4.9 1), 2.669 (0.43), 3.288 (0.67), 3.297 (0.72), 3.314 (0.92), 3.417 (0.41), 3.421 (1.02), 3.434 (1.07), 3.4 39 (1.07), 3.452 (1.13), 3.457 (0.54), 3.469 (0.53), 3.484 (0.62), 3.497 (0.71), 3.504 (0.71), 3.513 (0.72), 3.531 (1.07), 3.549 (0.56), 3.556 (0.49), 3.635 (0.82), 3.650 (0.97), 3.662 (0.83), 3.677 (0.77), 4.3 45 (1.31), 4.358 (1.96), 4.370 (0.96), 5.758 (0.45), 6.737 (3.67), 8.162 (1.02), 8.179 (1.01), 8.571 (4.12 ), 12.085 (0.80). Intermediate 10 N‐[(3S)‐1‐(4‐hydroxy‐2‐methylpyrido[3,4‐d]pyri midin‐6‐yl)pyrrolidin‐3‐yl]acetamide Analogously to Intermediate 9 using N‐[(3S)‐pyrrolidin‐3‐yl]acet amide (2.15 g, 16.7 mmol) gave the titled compound (1.06 g, 63%) after silica chromatogr aphy (DCM:EtOH). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.035 (2.38) , 1.052 (5.02), 1.069 (2.64), 1.807 (16.00), 1.898 (0.70), 1.912 (0.55), 2.159 (0.45), 2.174 (0.53), 2.1 90 (0.47), 2.283 (13.64), 2.518 (0.44), 3.287 (0.65), 3.297 (0.72), 3.314 (1.02), 3.337 (4.95), 3.428 (0.67), 3.445 (0.66), 3.482 (0.55), 3.495 (0.62), 3.502 (0.60), 3.513 (0.66), 3.547 (0.49), 3.555 (0.44), 3.6 34 (0.76), 3.649 (0.89), 3.660 (0.77), 3.676 (0.70), 4.347 (0.64), 4.361 (0.64), 5.758 (1.76), 6.732 (3.35), 6.734 (3.31), 8.161 (0.91), 8.177 (0.89), 8.567 (3.83), 8.568 (3.81). Intermediate 11
2‐methyl‐6‐(4‐methylpiperazin‐1‐yl)pyrido[3,4 ‐d]pyrimidin‐4‐ol Analogously to Intermediate 9 using 1‐methylpiperazine (2.24 g, 22.3 mmol) gave the titled compound (1.69 g, 55%) after silica chromatography (D CM:EtOH). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.052 (0.49) , 2.178 (0.53), 2.219 (13.17), 2.296 (16.00), 2.404 (2.87), 2.417 (3.99), 2.430 (3.10), 2.518 (1.22), 2.5 23 (0.83), 3.509 (2.77), 3.522 (3.47), 3.535 (2.74), 7.110 (3.66), 8.592 (4.09), 12.145 (0.89). Intermediate 12 6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐2‐me thylpyrido[3,4‐d]pyrimidin‐4‐ol Analogously to Intermediate 9 using (3R)‐N,N‐dimethylpyrrolidin‐3‐amine (2.55 g, 22.3 mmol) gave the titled compound (2.17 g, 68%) after silica chrom atography (DCM:EtOH). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.824 (0.41) , 2.206 (16.00), 2.279 (9.34), 2.288 (0.61), 3.131 (0.58), 3.152 (0.65), 3.155 (0.72), 3.176 (0.56), 3.3 64 (0.79), 3.381 (0.62), 3.390 (0.41), 3.619 (0.53), 3.694 (0.40), 3.712 (0.48), 3.719 (0.46), 6.751 (2.42 ), 6.753 (2.36), 8.555 (2.60), 8.557 (2.56). Intermediate 13 2‐(4‐hydroxy‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)‐2,6‐diazaspiro[3.4]octan‐7‐one Analogously to Intermediate 9 using 2,6‐diazaspiro[3. 4]octan‐7‐one oxalate salt (4.83 g, 22.3 mmol) gave the titled compound (1 g, 30%) after silica ch romatography (DCM:EtOH). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.035 (1.25) , 1.052 (2.83), 1.069 (1.11), 2.290 (16.00), 2.327 (0.45), 2.518 (2.47), 2.523 (1.44), 2.539 (8.49), 2.6 69 (0.48), 3.165 (6.18), 3.336 (0.51), 3.411 (0.68),
3.428 (1.27), 3.445 (1.25), 3.463 (0.57), 3.982 (1 5.13), 6.737 (4.83), 6.739 (4.60), 7.675 (1.53), 8.56 2 (4.66), 8.565 (4.55), 12.151 (0.73). Intermediate 14 1‐[4‐(4‐hydroxy‐2‐methylpyrido[3,4‐d]pyrimidin 6‐yl)piperazin‐1‐yl]ethan‐1‐one Analogously to Intermediate 9 using 1‐(piperazin‐1 yl)ethan‐1‐one (2.38 g, 18.6 mmol) gave the tit led compound (511 g, 16%) after silica chromatography (DC M:EtOH). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.052 (0.58) , 2.050 (16.00), 2.301 (14.44), 2.518 (1.23), 2.523 (0.89), 2.540 (2.37), 3.523 (1.33), 3.532 (1.43), 3.5 38 (1.95), 3.563 (3.09), 3.578 (3.59), 3.595 (2.31), 7.146 (3.41), 7.148 (3.39), 8.615 (3.86), 12.174 (0.8 4). Intermediate 15 7‐chloro‐2‐methylpyrido[4,3‐d]pyrimidin‐4‐ol To a solution of 5‐amino‐2‐chloropyridine‐4‐ca rboxylic acid (100 g, 579 mmol) and ethanimidamide hydrochloride (164 g, 1.74 mol) in 2‐methoxyethanol (1.2 L) was added sodium acetate (143 g, 1.74 mol) at room temperature. The reaction mixture was stirred at 130 °C for 48 hours. The reaction mixtrue was concentrated to remove about 400 ml 2‐ methoxyethanol under reduced pressure. The residue was poured into water, brown solid was precipitated. The precipi tates were filtered, dried under reduced pressure by oil pump to give 7‐chlor o‐2‐methylpyrido[4,3‐d]pyrimidin‐4‐ol as a brow n solid (77 g, 67%) 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 2.384 (1 6.00), 2.518 (0.89), 2.523 (0.59), 7.928 (4.21), 7.93 0 (4.17), 8.817 (3.76), 8.819 (3.55). Intermediate 16 1‐(4‐hydroxy‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)piperidine‐4‐carbonitrile
Analogously to Intermediate 9 using 1 piperidine‐4‐carbonitrile (2.46 g, 22.3 mmol) gave the titled compound (1.51 g, 48%) gave the title compound after silica chromatography (DCM:EtOH). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.052 (0.81), 1.070 (0.41), 1.722 (0.42), 1.734 (0.75), 1.744 (1.07), 1.755 (0.93), 1.767 (1.26), 1.776 (1.05), 1.7 89 (0.60), 1.798 (0.52), 1.919 (0.47), 1.928 (0.95), 1.936 (1.02), 1.944 (0.99), 1.952 (0.87), 1.961 (0.74 ), 1.968 (0.75), 1.976 (0.71), 2.296 (16.00), 2.518 (0.97), 2.523 (0.66), 3.114 (0.51), 3.124 (0.74), 3.1 34 (0.98), 3.145 (0.73), 3.156 (0.49), 3.393 (0.81), 3.401 (0.94), 3.414 (0.91), 3.426 (1.28), 3.435 (1.20), 3.448 (1.08), 3.456 (0.95), 3.820 (0.81), 3.830 (1.04), 3.836 (0.99), 3.846 (0.90), 3.854 (0.81), 3.8 63 (0.86), 3.870 (0.91), 3.879 (0.70), 5.758 (0.61), 7.157 (3.88), 8.599 (4.25), 12.156 (0.92). Intermediate 17 6‐[(2S)‐2,4‐dimethylpiperazin‐1‐yl]‐2‐methylpy rido[3,4‐d]pyrimidin‐4‐ol Analogously to Intermediate 9 using (3S)‐1,3‐dimeth ylpiperazine (153 mg, 1.34 mmol) gave the titled compound (30 mg, 16%) gave the title compound after preparative HPLC purification (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.120 (6.27), 1.137 (6.28), 1.751 (0.45), 1.921 (0.49), 1.941 (0.71), 1.950 (0.75), 1.971 (0.53), 1.979 (0.42), 2.1 14 (0.77), 2.125 (0.91), 2.142 (0.91), 2.152 (0.83), 2.201 (11.69), 2.291 (16.00), 2.304 (0.42), 2.518 (3. 07), 2.523 (2.26), 2.702 (1.07), 2.729 (0.99), 2.843 (0.68), 2.871 (0.64), 3.017 (0.43), 3.025 (0.49), 3.0 48 (0.91), 3.056 (0.83), 3.079 (0.52), 3.957 (0.61), 3.989 (0.57), 4.535 (0.56), 7.036 (3.56), 8.595 (3.91 ). Intermediate 18 6‐[2‐(hydroxymethyl)‐4‐methylpiperazin‐1‐yl]‐2 ‐methylpyrido[3,4‐d]pyrimidin‐4‐ol (mixture of stereoisomers)
Analogously to Intermediate 9 using [4‐methylpiperazi n‐2‐yl]methanol (218 mg, 1.67 mmol) gave the titled compound (40 mg, 17%) gave the title compound after preparative HPLC purification (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.751 (0.99), 1.909 (0.46), 1.930 (0.67), 1.938 (0.70), 1.960 (1.09), 1.969 (1.06), 1.988 (0.85), 1.998 (0.74), 2.0 66 (0.78), 2.118 (0.78), 2.200 (10.96), 2.287 (16.00) , 2.302 (0.49), 2.306 (0.46), 2.518 (3.49), 2.523 (2.57), 2.815 (0.67), 2.843 (0.60), 2.997 (0.46), 3.019 (0.81), 3.027 (0.74), 3.052 (1.30), 3.081 (0.95), 3.7 27 (0.49), 3.739 (0.49), 4.072 (0.56), 4.103 (0.53), 4.285 (0.53), 4.627 (0.42), 4.770 (0.60), 7.072 (3.49 ), 8.563 (4.23). Intermediate 19 2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐dihydroimida zo[1,2‐a]pyrazin‐7(8H)‐yl]pyrido[3,4‐d]pyrimidin‐4 ‐ ol Analogously to Intermediate 9 using 2‐(trifluoromethyl)‐5,6,7,8‐tetrahydroimidazo[1,2‐a] pyrazine (128 mg, 670 µmol) gave the title compound (40 mg, 34%) after preparative HPLC purification (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 2.315 (1 6.00), 2.327 (1.22), 2.332 (0.79), 2.518 (4.14), 2.52 3 (2.82), 2.665 (0.66), 2.669 (0.91), 2.673 (0.64), 4.1 52 (3.17), 4.162 (3.14), 4.821 (7.48), 7.335 (3.95), 7.805 (2.85), 7.808 (2.91), 8.673 (4.41). Intermediate 20 2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐dihydro[1,2, 4]triazolo[1,5‐a]pyrazin‐7(8H)‐yl]pyrido[3,4‐ d]pyrimidin‐4‐ol
Analogously to Intermediate 9 using 2‐(trifluoromethyl)‐5,6,7,8‐tetrahydro[1,2,4]triazolo[ 1,5‐ a]pyrazine (215 mg, 1.12 mmol) gave the title compound (25 mg, 13%) after preparative HPLC purification (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.056 (0 .83), 1.071 (0.88), 1.752 (0.45), 2.320 (16.00), 2.43 0 (0.80), 2.518 (7.83), 2.523 (5.50), 2.540 (1.66), 2.6 65 (0.77), 2.669 (1.03), 2.673 (0.77), 4.260 (1.23), 4.272 (2.59), 4.286 (2.00), 4.373 (1.76), 4.386 (2.40), 4.400 (1.13), 4.997 (6.61), 7.422 (3.99), 7.424 (3.99), 8.088 (0.45), 8.681 (4.22), 8.683 (4.22). Example 1 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ amine To a solution of 6‐ethoxy‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ol (75.0 mg, 365 µmol) and 2,4,6‐ tri(propan‐2‐yl)benzene‐1‐sulfonyl chloride (188 mg, 621 µmol) was added triethylamine (180 µl, 1.3 mmol) followed by DMAP (6.70 mg, 54.8 µmol) and st irred at RT for 1h. To the reaction was added (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐ 1‐amine hydrochloride (99.0 mg, 439 µmol) and stirred at RT overnight. The reaction was diluted wi th water and DCM and extracted with DCM. The organics were combined, washed with sat. NaCl(aq), filtered through an hydrophobic filter and concentrated under reduced pressure. The residue was purified by preparative HPLC (basic method) and gave the titled compound (14 mg, 10%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.349 (2.26), 1.366 (5.02), 1.384 (2.32), 1.586 (2.95), 1.604 (2.94), 2.322 (0.41), 2.326 (0.58), 2.331 (0.60), 2.3 42 (8.54), 2.518 (1.99), 2.522 (1.25), 2.669 (0.49), 4.337 (0.68), 4.355 (2.20), 4.372 (2.13), 4.389 (0.63 ), 5.742 (0.52), 5.758 (16.00), 5.777 (0.45), 7.098
(0.64), 7.234 (1.31), 7.268 (0.48), 7.287 (1.04), 7.306 (0.60), 7.370 (0.58), 7.502 (0.64), 7.667 (0.63 ), 7.745 (2.20), 8.567 (0.72), 8.585 (0.69), 8.698 (2.49 ). Example 2 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐fluoro‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ amine Using the method described for Example 1 using 6‐fluoro‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐ol (200 mg, 1.12 mmol) and (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐ 1‐amine hydrochloride (302 mg, 1.34 mmol) gave the titled compound (187 mg, 45 %) after preparative HPLC. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.604 (5.88) , 1.621 (5.85), 2.392 (16.00), 2.518 (1.16), 2.522 (0.75), 5.741 (0.80), 5.758 (2.24), 5.775 (0.79), 7.1 02 (1.27), 7.238 (2.56), 7.278 (0.89), 7.297 (1.99), 7.316 (1.13), 7.374 (1.16), 7.497 (0.70), 7.514 (1.18), 7.532 (0.57), 7.667 (0.64), 7.685 (1.18), 7.704 (0.58), 8.152 (2.56), 8.735 (3.89), 8.796 (1.23), 8.8 14 (1.20). Example 3 Example 3: N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluor omethyl)‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐ pyrido[3,4‐d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetam ide To a solution of Example 2 (40 mg, 114 µmol) in DMSO (1.5 ml) was added N‐[(3R)‐pyrrolidin‐3‐ yl]acetamide (58 mg, 457 µmol) and heated at 110°C overnight. The reaction was purified by preparative HPLC (basic method) and gave the titled compound (41 mg, 74%). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (1 .65), 1.603 (4.46), 1.620 (4.45), 1.826 (16.00), 1.93 2 (0.54), 1.945 (0.57), 2.183 (0.44), 2.199 (0.54), 2.2 14 (0.48), 2.290 (13.85), 2.332 (0.42), 2.518 (2.11), 2.523 (1.31), 2.673 (0.42), 3.302 (2.03), 3.312 (2.35), 3.328 (2.85), 3.339 (2.94), 3.504 (0.49), 3.518 (0.51), 3.525 (0.60), 3.530 (0.68), 3.538 (0.58), 3.5 44 (0.72), 3.550 (0.65), 3.563 (0.51), 3.601 (0.46), 3.620 (0.93), 3.638 (0.54), 3.646 (0.65), 3.665 (0.96), 3.681 (0.95), 3.693 (0.82), 3.708 (0.73), 4.395 (0.58), 4.407 (0.58), 5.762 (0.66), 5.780 (1.02), 5.7 98 (0.66), 7.079 (2.80), 7.101 (1.04), 7.237 (2.19),
7.276 (0.77), 7.295 (1.67), 7.314 (0.97), 7.373 (0.90), 7.483 (0.56), 7.501 (0.96), 7.518 (0.47), 7.629 (0.52), 7.647 (0.96), 7.665 (0.47), 8.155 (4.92), 8.1 96 (1.14), 8.212 (1.13), 8.395 (1.08), 8.414 (1.04), 8.633 (4.17). Example 4 N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 2 (40mg, 114 µmol) was treated with N‐[(3S)‐ pyrrolidin‐3‐yl]acetamide (59 mg, 457 µmol) and g ave the titled compound (41 mg, 75%). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (3 .59), 1.604 (4.53), 1.622 (4.52), 1.830 (16.00), 1.92 9 (0.56), 1.943 (0.58), 1.960 (0.42), 2.180 (0.46), 2.1 97 (0.55), 2.212 (0.49), 2.288 (14.00), 2.518 (1.56), 2.523 (0.97), 3.548 (0.41), 3.554 (0.59), 3.561 (0.43), 3.568 (0.67), 3.574 (0.63), 3.587 (0.86), 3.606 (0.98), 3.624 (0.53), 3.632 (0.54), 3.645 (0.83), 3.6 61 (0.93), 3.672 (0.79), 3.687 (0.71), 4.400 (0.58), 4.413 (0.58), 5.757 (0.70), 5.775 (1.05), 5.793 (0.67), 7.074 (2.82), 7.100 (1.06), 7.237 (2.21), 7.274 (0.78), 7.293 (1.70), 7.312 (0.99), 7.372 (0.91), 7.4 83 (0.57), 7.499 (0.98), 7.517 (0.48), 7.626 (0.52), 7.644 (0.96), 7.661 (0.48), 8.202 (1.23), 8.208 (0.94), 8.219 (1.21), 8.396 (1.15), 8.414 (1.11), 8.633 (4.25). Example 5 N‐[(1R)‐1‐[3‐(difluoromethyl)‐2‐fluoro‐phenyl] ethyl]‐2‐methyl‐6‐pyrrolidin‐1‐yl‐pyrido[3,4 d]pyrimidin‐4‐amine Using the method described for Example 3: Example 2 (40mg, 114 µmol) was treated with pyrrolidine (32 mg, 457 µmol) and gave the titled compound (42 mg, 86%). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (1 3.90), 1.604 (5.13), 1.622 (5.13), 1.990 (1.90), 2.00 0 (2.33), 2.007 (5.52), 2.014 (2.31), 2.023 (1.97), 2.2 85 (16.00), 2.518 (1.84), 2.522 (1.19), 3.448 (0.59), 3.457 (1.23), 3.473 (3.14), 3.481 (3.09), 3.497 (1.14), 3.506 (0.55), 5.762 (0.77), 5.780 (1.18), 5.798
(0.74), 7.061 (3.26), 7.100 (1.17), 7.236 (2.42), 7.270 (0.85), 7.289 (1.85), 7.308 (1.07), 7.372 (1.02 ), 7.481 (0.63), 7.498 (1.06), 7.516 (0.51), 7.634 (0.58), 7.652 (1.05), 7.670 (0.52), 8.362 (1.27), 8.380 (1.22), 8.621 (4.54). Example 6 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]eth yl}‐6‐fluoro‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ amine Using the method described for Example 1 using 6‐fluoro‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐ol and (1R)‐1‐[3‐(difluoromethyl)‐2‐methylphenyl]ethan‐ 1‐amine hydrochloride gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.542 (5 .50), 1.560 (5.58), 2.401 (16.00), 2.518 (1.42), 2.52 3 (1.00), 2.543 (8.10), 5.706 (0.82), 5.723 (1.27), 5.7 41 (0.81), 7.079 (1.03), 7.216 (2.15), 7.278 (0.70), 7.297 (1.70), 7.317 (1.11), 7.353 (0.92), 7.388 (1.66), 7.407 (1.12), 7.637 (1.34), 7.656 (1.19), 8.143 (2.44), 8.145 (2.48), 8.711 (4.26), 8.828 (1.24), 8.8 46 (1.20). Example 7 N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 6 was treated with N‐[(3R)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.539 (3 .55), 1.556 (3.58), 1.825 (13.28), 1.928 (0.46), 1.94 2 (0.48), 2.195 (0.47), 2.210 (0.41), 2.303 (11.17), 2. 323 (0.45), 2.327 (0.57), 2.518 (2.39), 2.523 (1.71), 2.540 (16.00), 2.669 (0.52), 3.300 (0.57), 3.310 (0.6 6), 3.523 (0.45), 3.536 (0.47), 3.542 (0.42), 3.613 (0.72), 3.631 (0.40), 3.638 (0.50), 3.658 (0.77), 3.6 74 (0.76), 3.685 (0.64), 3.700 (0.57), 4.391 (0.49), 4.405 (0.48), 5.720 (0.55), 5.738 (0.84), 5.756 (0.54), 7.069 (2.40), 7.075 (0.97), 7.214 (1.47), 7.277
(0.52), 7.296 (1.27), 7.315 (0.84), 7.351 (0.62), 7.376 (1.27), 7.393 (0.82), 7.630 (1.01), 7.649 (0.89 ), 8.192 (0.97), 8.208 (0.96), 8.434 (0.95), 8.453 (0.91 ), 8.610 (3.38). Example 8 N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(difluoromethyl)‐2 methyl‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 6 was treated with N‐[(3S)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.539 (4 .28), 1.557 (4.28), 1.829 (16.00), 1.925 (0.58), 1.93 8 (0.59), 1.956 (0.41), 2.175 (0.46), 2.191 (0.55), 2.2 06 (0.49), 2.303 (13.86), 2.323 (0.54), 2.327 (0.69), 2.331 (0.49), 2.518 (2.77), 2.523 (1.97), 2.541 (6.90), 2.665 (0.47), 2.669 (0.63), 2.673 (0.44), 3.308 (0.74), 3.319 (0.95), 3.345 (0.98), 3.550 (0.57), 3.5 64 (0.65), 3.570 (0.63), 3.582 (0.75), 3.599 (0.98), 3.616 (0.51), 3.625 (0.50), 3.638 (0.81), 3.654 (0.91), 3.665 (0.76), 3.680 (0.67), 4.400 (0.59), 4.413 (0.57), 5.716 (0.66), 5.734 (1.01), 5.751 (0.65), 7.0 63 (2.77), 7.076 (0.90), 7.214 (1.77), 7.275 (0.64), 7.294 (1.53), 7.313 (1.04), 7.351 (0.73), 7.375 (1.50), 7.393 (0.97), 7.627 (1.19), 7.646 (1.06), 8.199 (1.19), 8.216 (1.15), 8.435 (1.12), 8.453 (1.07), 8.6 10 (4.01). Example 9 N‐[(1R)‐1‐[3‐(difluoromethyl)‐2‐methyl‐phenyl] ethyl]‐2‐methyl‐6‐pyrrolidin‐1‐yl‐pyrido[3,4 d]pyrimidin‐4‐amine Using the method described for Example 3: Example 6 was treated with pyrrolidine and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.541 (5.03), 1.558 (4.97), 1.986 (1.89), 1.996 (2.29), 2.003 (5.67), 2.010 (2.30), 2.019 (1.99), 2.299 (16.00), 2. 322 (0.61), 2.326 (0.81), 2.332 (0.59), 2.518 (3.54),
2.523 (2.38), 2.539 (7.68), 2.664 (0.57), 2.669 (0.80), 2.673 (0.58), 3.441 (0.58), 3.450 (1.11), 3.466 (3.01), 3.476 (3.00), 3.483 (1.78), 3.492 (1.08), 3.5 02 (0.57), 5.720 (0.73), 5.739 (1.14), 5.756 (0.73), 7.051 (3.18), 7.075 (0.97), 7.212 (2.01), 7.271 (0.67), 7.290 (1.65), 7.310 (1.10), 7.350 (0.84), 7.374 (1.62), 7.392 (1.06), 7.640 (1.28), 7.660 (1.14), 8.3 96 (1.23), 8.414 (1.19), 8.599 (4.28). Example 10 6‐fluoro‐2‐methyl‐N‐[(1R)‐1‐[3‐(trifluoromet hyl)phenyl]ethyl]pyrido[3,4‐d]pyrimidin‐4‐amine Using the method described for Example 1 using 6‐fluoro‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐ol and (1R)‐1‐[3‐(trifluoromethyl)phenyl]ethan‐1‐amine h ydrochloride gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.612 (5 .87), 1.629 (5.90), 1.986 (0.64), 2.421 (16.00), 2.51 8 (1.36), 2.523 (0.91), 5.603 (0.79), 5.620 (1.18), 5.6 39 (0.77), 7.550 (0.50), 7.569 (1.59), 7.588 (1.81), 7.598 (2.01), 7.617 (0.61), 7.752 (1.37), 7.770 (1.08), 7.830 (2.25), 8.098 (2.53), 8.101 (2.53), 8.731 (4.14), 8.765 (1.20), 8.784 (1.16). Example 11 N‐[(3R)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluo romethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐6‐ yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 10 was treated with N‐[(3R)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.613 (4 .68), 1.631 (4.71), 1.823 (16.00), 1.927 (0.53), 1.94 0 (0.56), 2.179 (0.44), 2.195 (0.52), 2.210 (0.46), 2.3 21 (14.30), 2.332 (0.69), 2.518 (1.70), 2.523 (1.16), 2.669 (0.57), 3.294 (0.64), 3.303 (0.69), 3.519 (0.40), 3.525 (0.50), 3.532 (0.40), 3.538 (0.55), 3.545 (0.49), 3.610 (0.90), 3.628 (0.48), 3.636 (0.59), 3.6 55 (0.91), 3.670 (0.88), 3.682 (0.76), 3.698 (0.67), 4.390 (0.56), 4.404 (0.55), 5.623 (0.67), 5.642 (0.98), 5.659 (0.64), 7.033 (2.77), 7.569 (1.35), 7.589
(2.87), 7.732 (1.17), 7.750 (0.87), 7.797 (1.99), 8.191 (1.11), 8.208 (1.09), 8.377 (1.14), 8.397 (1.10 ), 8.633 (4.11). Example 12 N‐[(3S)‐1‐[2‐methyl‐4‐[[(1R)‐1‐[3‐(trifluo romethyl)phenyl]ethyl]amino]pyrido[3,4‐d]pyrimidin‐6‐ yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 10 was treated with N‐[(3S)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.615 (4 .44), 1.633 (4.45), 1.824 (16.00), 1.924 (0.53), 1.93 8 (0.54), 2.176 (0.43), 2.193 (0.51), 2.207 (0.44), 2.3 20 (13.83), 2.332 (0.62), 2.518 (1.81), 2.523 (1.17), 2.540 (5.21), 2.669 (0.52), 3.304 (0.67), 3.315 (0.78), 3.546 (0.53), 3.559 (0.59), 3.565 (0.55), 3.580 (0.68), 3.599 (0.90), 3.618 (0.48), 3.625 (0.51), 3.6 36 (0.76), 3.651 (0.87), 3.662 (0.73), 3.678 (0.64), 4.395 (0.54), 4.409 (0.53), 5.618 (0.63), 5.636 (0.93), 5.654 (0.61), 7.027 (2.64), 7.568 (1.28), 7.587 (2.70), 7.592 (1.40), 7.732 (1.12), 7.749 (0.83), 7.7 95 (1.87), 8.196 (1.09), 8.213 (1.08), 8.376 (1.13), 8.395 (1.07), 8.634 (3.91). Example 13 N‐[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl]‐6 fluoro‐2‐methyl‐pyrido[3,4‐d]pyrimidin‐4‐amine Using the method described for Example 1 using 6‐fluoro‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐ol and (1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethanamine hydr ochloride gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.600 (5.73), 1.618 (5.72), 1.910 (4.85), 1.957 (9.96), 2.004 (4.33), 2.435 (16.00), 2.518 (2.83), 2.523 (1.94), 2. 673 (0.48), 5.596 (0.56), 5.614 (0.85), 5.632 (0.55), 7.436 (3.99), 7.442 (1.34), 7.451 (1.86), 7.471 (0.45), 7.567 (1.06), 7.579 (0.73), 7.583 (0.95), 7.672 (2.13), 8.110 (2.28), 8.113 (2.30), 8.726 (3.70), 8.7 38 (0.94), 8.756 (0.86).
Example 14 N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐d]pyrimidin 6‐ yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 13 was treated with N‐[(3R)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.600 (4 .60), 1.618 (4.63), 1.822 (16.00), 1.906 (4.45), 1.92 4 (0.60), 1.937 (0.69), 1.953 (8.68), 2.000 (3.68), 2.1 75 (0.50), 2.192 (0.57), 2.207 (0.51), 2.223 (0.41), 2.323 (1.11), 2.333 (14.89), 2.518 (4.14), 2.523 (2.8 1), 2.665 (0.69), 2.669 (0.96), 2.673 (0.68), 3.291 (0.57), 3.301 (0.75), 3.318 (0.95), 3.516 (0.41), 3.5 22 (0.48), 3.535 (0.57), 3.542 (0.56), 3.555 (0.47), 3.606 (0.81), 3.614 (0.47), 3.624 (0.51), 3.632 (0.68), 3.649 (0.93), 3.664 (0.81), 3.676 (0.77), 3.692 (0.60), 4.391 (0.60), 4.403 (0.59), 5.619 (0.60), 5.6 37 (0.87), 5.656 (0.57), 7.042 (2.54), 7.407 (0.45), 7.429 (2.75), 7.447 (1.71), 7.466 (0.59), 7.547 (1.22), 7.564 (0.99), 7.651 (2.18), 8.191 (1.10), 8.208 (1.10), 8.353 (1.23), 8.372 (1.17), 8.628 (4.36). Example 15 N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) phenyl]ethyl]amino]‐2‐methyl‐pyrido[3,4‐d]pyrimidin 6‐ yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 13 was treated with N‐[(3S)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.602 (4 .60), 1.619 (4.59), 1.822 (16.00), 1.904 (4.38), 1.92 1 (0.62), 1.935 (0.66), 1.952 (8.60), 1.998 (3.64), 2.1 73 (0.54), 2.190 (0.59), 2.206 (0.51), 2.333 (14.50), 2.518 (3.18), 2.523 (2.14), 2.665 (0.53), 2.669 (0.73), 2.673 (0.52), 3.301 (0.73), 3.311 (0.72), 3.535 (0.46), 3.541 (0.59), 3.554 (0.62), 3.561 (0.52), 3.5 75 (0.53), 3.597 (0.82), 3.605 (0.47), 3.615 (0.53),
3.623 (0.56), 3.631 (0.81), 3.647 (0.92), 3.658 (0.70), 3.674 (0.69), 4.393 (0.61), 4.405 (0.60), 5.614 (0.59), 5.632 (0.88), 5.651 (0.60), 7.038 (2.54), 7.4 06 (0.46), 7.428 (2.78), 7.446 (1.74), 7.466 (0.59), 7.547 (1.21), 7.564 (0.99), 7.650 (2.20), 8.195 (1.12), 8.211 (1.10), 8.352 (1.22), 8.372 (1.17), 8.628 (4.42). Example 16 N‐[(1R)‐1‐[3‐(1,1‐difluoroethyl)phenyl]ethyl]‐6 fluoro‐2‐methyl‐pyrido[3,4‐d]pyrimidin‐4‐amine Using the method described for Example 1 using 6‐fluoro‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐ol and (1R)‐1‐[3‐(1,1‐difluoroethyl)‐2‐fluoro‐phenyl] ethanamine hydrochloride gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.600 (5.37), 1.617 (5.29), 1.981 (2.65), 2.028 (5.15), 2.076 (2.38), 2.332 (0.68), 2.392 (16.00), 2.518 (3.61), 2. 523 (2.50), 2.673 (0.68), 5.735 (0.79), 5.753 (1.23), 5.770 (0.79), 7.235 (0.82), 7.254 (1.84), 7.273 (1.08), 7.429 (0.66), 7.447 (1.11), 7.463 (0.56), 7.621 (0.59), 7.637 (1.05), 7.655 (0.52), 8.150 (2.33), 8.1 53 (2.33), 8.735 (3.90), 8.792 (1.14), 8.810 (1.11). Example 17 N‐[(3R)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) ‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3, 4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 16 was treated with N‐[(3R)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.597 (4 .59), 1.615 (4.59), 1.827 (16.00), 1.932 (0.59), 1.94 5 (0.64), 1.963 (0.51), 1.981 (2.75), 2.029 (5.16), 2.0 77 (2.35), 2.183 (0.54), 2.199 (0.60), 2.214 (0.54), 2.232 (0.43), 2.289 (13.51), 2.518 (4.05), 2.523 (2.7 2), 3.301 (0.73), 3.312 (0.87), 3.526 (0.44), 3.532 (0.56), 3.545 (0.60), 3.551 (0.52), 3.621 (0.89), 3.6 39 (0.51), 3.647 (0.62), 3.667 (0.92), 3.683 (0.94),
3.694 (0.81), 3.710 (0.71), 4.395 (0.62), 4.408 (0.60), 5.755 (0.73), 5.773 (1.11), 5.791 (0.70), 7.079 (2.91), 7.231 (0.83), 7.250 (1.80), 7.269 (1.05), 7.4 13 (0.67), 7.431 (1.08), 7.447 (0.51), 7.583 (0.59), 7.600 (1.05), 7.617 (0.52), 8.198 (1.22), 8.214 (1.18 ), 8.396 (1.18), 8.414 (1.13), 8.630 (4.27). Example 18 N‐[(3S)‐1‐[4‐[[(1R)‐1‐[3‐(1,1‐difluoroethyl) ‐2‐fluoro‐phenyl]ethyl]amino]‐2‐methyl‐pyrido[3, 4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl]acetamide Using the method described for Example 3: Example 16 was treated with N‐[(3S)‐pyrrolidin‐3‐ yl]acetamide and gave the titled compound. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.514 (0 .42), 1.598 (4.69), 1.616 (4.66), 1.830 (16.00), 1.91 2 (0.41), 1.930 (0.64), 1.944 (0.67), 1.960 (0.65), 1.9 81 (2.81), 2.007 (0.47), 2.029 (5.36), 2.076 (2.45), 2.182 (0.56), 2.197 (0.67), 2.216 (0.92), 2.229 (0.52 ), 2.287 (13.61), 2.318 (0.44), 2.323 (0.80), 2.327 (1.08), 2.331 (0.78), 2.518 (3.87), 2.523 (2.53), 2.6 65 (0.67), 2.669 (0.95), 2.673 (0.64), 3.315 (1.05), 3.352 (0.80), 3.556 (0.59), 3.570 (0.69), 3.576 (0.64), 3.589 (0.87), 3.606 (1.03), 3.624 (0.54), 3.632 (0.52), 3.646 (0.85), 3.662 (0.95), 3.673 (0.80), 3.6 88 (0.70), 4.401 (0.62), 4.413 (0.60), 5.750 (0.74), 5.767 (1.13), 5.785 (0.70), 7.074 (2.93), 7.229 (0.85), 7.248 (1.85), 7.267 (1.11), 7.412 (0.67), 7.430 (1.09), 7.447 (0.52), 7.580 (0.62), 7.596 (1.06), 7.6 14 (0.52), 8.203 (1.26), 8.220 (1.21), 8.396 (1.21), 8.414 (1.14), 8.631 (4.33). Example 19 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐fluoro‐2,8‐dimethylpyrido[3,4‐d]pyrimidin‐ 4‐ amine
To a solution of Example 2 (250 mg, 714 µmol) in DMSO (5 ml) was added DBU (213 µl, 3.6 mmol) and nitromethane (193 µl, 1.43 mmol) and stirred for 4 days at RT. The reaction was diluted with water and the solid collected by filtration and wash ed with water. The solid was dried to give the tit le compound (261 mg, 95%). ¹H‐NMR (600 MHz, DMSO‐d6) δ [ppm]: 0.909 (0.44), 1.111 (2.03), 1.233 (0.43), 1.601 (6.17), 1.612 (5.96), 2.386 (0.69), 2.388 (0.89), 2.391 (0.77), 2.3 95 (0.65), 2.403 (16.00), 2.519 (1.95), 2.522 (1.82), 2.525 (1.44), 2.613 (0.46), 2.616 (0.66), 2.619 (0.53 ), 2.727 (12.15), 3.313 (0.74), 5.757 (0.60), 7.142 (1.06), 7.232 (2.12), 7.276 (0.96), 7.289 (2.03), 7.3 02 (1.12), 7.323 (0.94), 7.496 (0.63), 7.508 (1.10), 7.519 (0.57), 7.658 (0.60), 7.669 (1.12), 7.681 (0.57), 7.949 (2.43), 8.088 (0.78), 8.316 (4.63), 8.693 (0.48). Example 20 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide To a solution of Example 19 (20.8 mg, 57 µmol) in DMSO (0.5 ml) was added N‐[(3R)‐pyrrolidin‐3‐ yl]acetamide (14 mg, 114 µmol) and TEA (32 µl, 22 8 µmol). The reaction was heated at 110°C for 16h . The reaction was allowed to cool and then purified by preparative HPLC (basic method) to give the titled compound (9.5 mg, 35%). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.094 (3.50), 1.170 (0.41), 1.228 (1.01), 1.591 (5.83), 1.608 (6.19), 1.820 (16.00), 1.903 (1.21), 1.913 (0.98), 1. 927 (0.99), 1.944 (0.74), 2.164 (0.77), 2.179 (0.98), 2.195 (0.91), 2.211 (0.68), 2.297 (12.97), 2.323 (1.1 9), 2.637 (13.81), 2.657 (1.41), 2.665 (1.17), 3.286 (1.41), 3.297 (1.92), 3.478 (0.48), 3.503 (0.95), 3.5 17 (1.03), 3.536 (0.66), 3.589 (0.59), 3.606 (1.24), 3.624 (0.90), 3.631 (1.01), 3.655 (1.28), 3.670 (1.35), 3.682 (1.17), 3.698 (1.01), 4.352 (0.62), 4.366 (1.06), 4.379 (1.05), 5.753 (0.98), 5.770 (1.50), 5.7 88 (1.01), 6.896 (3.50), 7.095 (1.23), 7.231 (2.49), 7.264 (1.09), 7.283 (2.32), 7.302 (1.39), 7.367 (1.13), 7.473 (0.99), 7.490 (1.63), 7.507 (0.88), 7.614 (0.92), 7.632 (1.64), 7.649 (0.87), 8.084 (0.50), 8.1 72 (1.75), 8.188 (1.72), 8.275 (1.03), 8.292 (1.07). Example 21 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[(3R)‐3‐(dimethylamino)pyrrolidin‐1‐yl]‐ 2,8‐ dimethylpyrido[3,4‐d]pyrimidin‐4‐amine
Using the method described for Example 20: Example 19 was treated with (3R)‐N,N‐ dimethylpyrrolidin‐3‐amine (58.0 mg, 508 µmol) and gave the titled compound (25 mg, 51%) after preparative HPLC (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.602 (3.56), 1.619 (3.58), 1.854 (0.51), 1.876 (0.42), 2.182 (0.49), 2.197 (0.49), 2.239 (16.00), 2.296 (9.36), 2. 323 (0.67), 2.327 (0.82), 2.639 (8.90), 2.665 (0.68), 2.669 (0.81), 2.812 (0.40), 2.830 (0.53), 3.153 (0.62), 3.178 (0.80), 3.198 (0.59), 3.383 (0.67), 3.400 (0.64), 3.645 (0.43), 3.666 (0.70), 3.742 (0.56), 3.7 59 (0.67), 3.766 (0.64), 3.784 (0.48), 5.757 (0.56), 5.775 (0.86), 5.793 (0.54), 6.865 (2.29), 7.100 (0.76), 7.236 (1.58), 7.262 (0.61), 7.282 (1.35), 7.301 (0.80), 7.371 (0.70), 7.474 (0.53), 7.491 (0.87), 7.5 09 (0.44), 7.619 (0.48), 7.638 (0.87), 7.655 (0.45), 8.217 (0.95), 8.235 (0.94). Example 22 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐d]pyrimidi n‐ 6‐yl]piperazin‐1‐yl}ethan‐1‐one Using the method described for Example 20: Example 1 9 was treated with 1‐(piperazin‐1‐yl)ethan‐1‐ one (65.1 mg, 508 µmol) and gave the titled compou nd (20 mg, 40%) after preparative HPLC (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.967 (0.44), 1.107 (0.42), 1.603 (5.55), 1.621 (5.45), 1.957 (0.40), 2.074 (16.00), 2.321 (15.94), 2.432 (0.46), 2.522 (4.88), 2.658 (14.29), 2.669 (1.88), 3.516 (1.82), 3.606 (9.65), 5.749 (0.91), 5.766 (1.31), 5.7 84 (0.82), 7.101 (1.29), 7.238 (2.64), 7.272 (0.97), 7.293 (4.58), 7.310 (1.29), 7.374 (1.12), 7.485 (0.72), 7.500 (1.25), 7.517 (0.63), 7.622 (0.70), 7.641 (1.22), 7.658 (0.63), 8.340 (1.37), 8.359 (1.35). Example 23
N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl] ethyl}‐2,8‐dimethyl‐6‐(4‐methylpiperazin‐1‐ yl)pyrido[3,4‐d]pyrimidin‐4‐amine Using the method described for Example 20: Example 1 9 was treated with 1‐methylpiperazine (110 µl, 1.0 mmol) and gave the titled compound (30 mg, 60%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.860 (0.75), 0.967 (2.61), 1.109 (1.08), 1.144 (1.52), 1.209 (0.57), 1.224 (0.66), 1.596 (5.12), 1.614 (5.11), 2.2 52 (10.44), 2.313 (16.00), 2.322 (1.22), 2.327 (1.13) , 2.332 (0.78), 2.459 (2.46), 2.471 (3.88), 2.518 (3.87 ), 2.523 (2.45), 2.642 (13.95), 2.660 (0.42), 2.665 (0.72), 2.669 (0.97), 2.673 (0.71), 3.525 (2.34), 3.5 37 (3.14), 3.549 (2.33), 5.744 (0.78), 5.762 (1.20), 5.780 (0.77), 7.101 (1.16), 7.237 (2.68), 7.245 (3.17), 7.269 (0.89), 7.289 (1.92), 7.307 (1.11), 7.373 (1.02), 7.480 (0.65), 7.497 (1.10), 7.514 (0.54), 7.6 20 (0.59), 7.637 (1.08), 7.655 (0.53), 8.313 (1.29), 8.331 (1.24). Example 24 2‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophen yl]ethyl}amino)‐2,8‐dimethylpyrido[3,4‐d]pyrimidin‐6 ‐ yl]‐2,6‐diazaspiro[3.4]octan‐7‐one Using the method described for Example 20: Example 1 9 was treated with 2,6‐diazaspiro[3.4]octan‐ 7‐one (64.1 mg, 508 µmol) and gave the titled co mpound (20 mg, 40%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.109 (0.49), 1.231 (0.52), 1.348 (0.42), 1.569 (0.44), 1.587 (5.53), 1.605 (5.56), 2.286 (0.50), 2.310 (16.00), 2. 322 (1.35), 2.327 (1.54), 2.332 (1.12), 2.422 (0.79), 2.428 (0.51), 2.432 (0.74), 2.449 (0.49), 2.518 (5.90 ), 2.523 (3.76), 2.542 (8.45), 2.632 (13.73), 2.660 (0.51), 2.665 (1.05), 2.669 (1.45), 2.673 (1.05), 2.6 78 (0.49), 3.522 (6.98), 3.954 (0.86), 3.978 (8.61), 4.003 (0.84), 5.738 (0.91), 5.756 (1.33), 5.774 (0.83), 6.966 (3.70), 7.097 (1.28), 7.233 (2.68), 7.265
(0.95), 7.285 (2.08), 7.303 (1.25), 7.369 (1.15), 7.478 (0.74), 7.495 (1.24), 7.513 (0.64), 7.617 (0.69 ), 7.634 (1.30), 7.653 (0.69), 7.676 (2.61), 8.088 (0.56 ), 8.299 (1.40), 8.317 (1.35). Example 25 N‐{(3S)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)pheny l]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]pyrrolidin‐3‐yl}acetamide To a solution of intermediate 10 (57.7 mg, 201 µmo l) and PyBOP (136 mg, 261 µmol) in DMF (580 µL) was added DBU (90 µl, 600 µmol) followed by (1R)‐1‐[3‐(difluoromethyl)phenyl]ethan‐1‐amine hydrochloride (50.0 mg, 241 µmol). The reaction was stirred at RT for 16h. The titled compound was isolated (50 mg, 54%) after preparative HPLC purifica tion (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.601 (4.90) , 1.619 (4.90), 1.823 (16.00), 1.922 (0.60), 1.935 (0.61), 1.953 (0.44), 2.174 (0.49), 2.190 (0.58), 2.2 05 (0.52), 2.221 (0.40), 2.326 (14.40), 2.518 (1.83), 2.522 (1.14), 2.669 (0.42), 3.303 (0.74), 3.313 (0.85), 3.542 (0.59), 3.550 (0.43), 3.556 (0.67), 3.563 (0.62), 3.577 (0.71), 3.597 (1.00), 3.615 (0.54), 3.6 23 (0.59), 3.632 (0.84), 3.648 (0.99), 3.658 (0.81), 3.674 (0.72), 4.394 (0.62), 4.406 (0.61), 5.623 (0.67), 5.641 (1.01), 5.659 (0.66), 6.884 (1.24), 7.024 (2.60), 7.045 (2.98), 7.164 (1.15), 7.415 (0.75), 7.4 34 (1.54), 7.459 (1.15), 7.478 (1.73), 7.497 (0.74), 7.597 (1.22), 7.616 (0.96), 7.637 (2.00), 8.195 (1.21), 8.211 (1.20), 8.361 (1.23), 8.381 (1.18), 8.630 (4.29). Example 26 N‐{(3S)‐1‐[2‐methyl‐4‐({(1R)‐1‐[2‐methyl 3‐(trifluoromethyl)phenyl]ethyl}amino)pyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}acetamide Using the method described for Example 25: Intermedia te 10 was treated with (1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethan‐1‐amine hydrochloride (50.0 mg, 209 µmol) and gave the titled compound (30 mg, 35%) after preparative HPLC purifica tion (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.557 (4.32) , 1.575 (4.42), 1.831 (16.00), 1.928 (0.56), 1.941 (0.56), 1.960 (0.41), 2.178 (0.45), 2.194 (0.54), 2.2 09 (0.49), 2.226 (0.40), 2.281 (13.18), 2.518 (3.92), 2.523 (2.45), 2.539 (0.42), 2.618 (5.45), 3.313 (0.80), 3.349 (1.00), 3.554 (0.55), 3.567 (0.64), 3.574 (0.62), 3.585 (0.71), 3.602 (0.99), 3.620 (0.51), 3.6 28 (0.49), 3.643 (0.82), 3.658 (0.90), 3.670 (0.76), 3.685 (0.68), 4.402 (0.56), 4.414 (0.56), 5.682 (0.65), 5.699 (1.02), 5.716 (0.66), 7.062 (2.74), 7.338 (0.63), 7.357 (1.37), 7.377 (0.80), 7.527 (1.47), 7.5 46 (1.20), 7.737 (1.29), 7.756 (1.17), 8.202 (1.19), 8.218 (1.14), 8.492 (1.11), 8.510 (1.07), 8.613 (4.06 ). Table 1: Examples 27‐34 Using the method described for Example 25: Intermediate 7 was treated with the corresponding phenylethan‐1‐amines or their hydrochloride salts a nd gave the desired compounds after preparative HPLC purification (basic method).
Table 2: Examples 35‐42 Using the method described for Example 25: Intermediate 8 was treated with the corresponding phenylethan‐1‐amines or their hydrochloride salts a nd gave the desired compounds after preparative HPLC purification (basic method).
Table 3: Examples 43‐91 Using the method described for Example 25: Intermediate 9 was treated with the corresponding phenylethan‐1‐amines or their hydrochloride salts a nd gave the desired compounds after preparative HPLC purification (basic method).
Example 92 N‐[(3R)‐1‐(4‐{[1‐(3‐aminophenyl)ethyl]amino}‐2 ‐methylpyrido[3,4‐d]pyrimidin‐6‐yl)pyrrolidin‐3‐ yl]acetamide (mixture of stereoisomers) To Example 68 (25.0 mg, 49.4 µmol) was added 4M H Cl in dioxane (3.1 ml) followed by MeOH (3 ml). The reaction was stirred at RT for 3h and concentrated. The titled compound (9 mg, 43%) was isolated after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.514 (4.94) , 1.532 (5.01), 1.819 (16.00), 1.914 (0.59), 1.928 (0.64), 1.944 (0.49), 2.167 (0.57), 2.184 (0.74), 2.1 99 (0.62), 2.215 (0.48), 2.327 (0.77), 2.344 (15.22), 2.522 (2.32), 2.665 (0.44), 2.669 (0.61), 2.673 (0.44), 3.280 (0.42), 3.290 (0.54), 3.305 (0.88), 3.528 (0.62), 3.545 (0.57), 3.590 (0.66), 3.603 (0.73), 3.6 22 (0.79), 3.638 (0.76), 3.659 (0.70), 3.669 (0.61), 3.685 (0.47), 4.370 (0.45), 4.384 (0.77), 4.397 (0.73), 4.410 (0.44), 5.003 (3.73), 5.512 (0.68), 5.531 (0.99), 5.548 (0.68), 6.392 (1.18), 6.396 (1.20), 6.4 12 (1.25), 6.415 (1.32), 6.563 (1.45), 6.583 (1.90), 6.589 (2.20), 6.594 (2.57), 6.931 (1.52), 6.950 (2.59), 6.970 (1.28), 7.070 (2.87), 8.184 (1.30), 8.201 (1.32), 8.219 (1.48), 8.239 (1.40), 8.617 (5.00). Example 93 tert‐butyl {3‐[(1S)‐1‐({6‐[(3R)‐3‐acetamidopy rrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4 yl}amino)ethyl]phenyl}carbamate Example 68 (116 mg, 230 µmol) was purified by chir al HPLC to give: Example 93 (68 mg, 56%, e.e. >95%). Rt = 8.51 m in Example 94 (37 mg, 33%, e.e. >95%). Rt = 6.24 m ins Analytical Method: Instrument: Agilent: 1260, Aurora SFC‐Module; Column: Chiralpak IC 5µ 100x4.6mm; eluent A: CO2; eluent B: 2‐propanol + 0 .4 vol % diethylamine; isocratic: 30%B; flow: 4 ml/min; temperature: 37.5°C; BPR: 100bar; UV: 280 nm
Preparative Method: Instrument: Sepiatec: Prep SFC10 0; Column: Chiralpak IC 5µ 250x30mm; eluent A: CO2; eluent B: 2‐propanol + 0.4 vol % diethylamine; isocratic: 30%B; flow: 100 ml/min; temperature: 40°C; BPR: 150bar; UV: 280 nm. Example 94 tert‐butyl {3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopy rrolidin‐1‐yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4 yl}amino)ethyl]phenyl}carbamate See example 93 for details. Example 95 N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3‐aminophenyl)ethyl]ami no}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl)pyrrolidin ‐3‐ yl]acetamide Using the method described for Example 92: Example 93 gave the titled compound (12 mg, 60%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.593 (4.29) , 1.610 (4.27), 1.822 (16.00), 1.949 (0.54), 1.963 (0.56), 1.979 (0.47), 2.190 (0.41), 2.206 (0.52), 2.2 22 (0.49), 2.518 (3.31), 2.523 (2.89), 2.530 (9.56), 3.316 (0.90), 3.326 (1.04), 3.344 (1.36), 3.558 (0.52), 3.572 (0.58), 3.592 (0.67), 3.611 (0.74), 3.629 (0.46), 3.637 (0.44), 3.655 (0.79), 3.670 (0.79), 3.6 81 (0.67), 3.697 (0.59), 4.380 (0.60), 4.392 (0.59), 5.665 (0.56), 5.684 (0.80), 5.701 (0.55), 6.468 (0.91), 6.471 (0.93), 6.487 (0.96), 6.490 (1.01), 6.590 (1.05), 6.609 (1.26), 6.619 (1.54), 6.623 (1.93), 6.9 88 (1.36), 7.007 (2.26), 7.026 (1.16), 7.276 (2.17), 8.209 (1.18), 8.225 (1.15), 8.726 (4.11). Example 96 N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐aminophenyl)ethyl]ami no}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl)pyrrolidin ‐3‐ yl]acetamide
Using the method described for Example 92: Example 94 gave the titled compound (12 mg, 60%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.584 (4.03) , 1.601 (4.08), 1.821 (16.00), 1.950 (0.51), 1.962 (0.55), 1.980 (0.44), 2.204 (0.49), 2.219 (0.44), 2.3 23 (0.51), 2.327 (0.73), 2.332 (0.52), 2.518 (3.95), 2.523 (2.49), 2.665 (0.53), 2.669 (0.76), 2.673 (0.52), 3.301 (0.83), 3.311 (1.01), 3.537 (0.47), 3.551 (0.50), 3.622 (0.67), 3.640 (0.40), 3.648 (0.46), 3.6 70 (0.75), 3.686 (0.80), 3.698 (0.69), 3.713 (0.60), 4.379 (0.57), 4.392 (0.56), 5.650 (0.52), 5.668 (0.75), 5.687 (0.50), 6.456 (0.90), 6.460 (0.90), 6.476 (0.94), 6.480 (0.99), 6.583 (1.08), 6.603 (1.29), 6.6 13 (1.51), 6.617 (1.96), 6.981 (1.37), 7.000 (2.27), 7.019 (1.17), 7.263 (1.88), 8.205 (1.10), 8.222 (1.07 ), 8.716 (3.74). Example 97 N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3,5‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide Example 73 (70 mg, 188 µmol) was purified by chira l HPLC to give: Example 97 (25 mg, e.e. >95%). Rt = 4.57 min Example 98 (23 mg, e.e. >95%). Rt = 5.44 mins Analytical Method: Instrument: Thermo Fisher UltiMate 3000; Column: YMC Cellulose SB 3µ, 100x4.6; eluent A: methyl tert‐butyl ether + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 95%A+5%B; flow: 1.4 ml/min; temperature: 25°C; UV: 2 80 nm Preparative Method: Instrument: PrepCon Labomatic HPLC‐3; Column: YMC Cellulose SB 10µ, 250x50; eluent A: methyl tert‐butyl ether + 0.1 vol % diethylamine; eluent B: ethanol + 0.1 vol % diethylamine; isocratic: 95%A+5%B; flow: 80 ml/min; te mperature: 25°C; UV: 280 nm ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.570 (4.93) , 1.588 (5.00), 1.823 (16.00), 1.928 (0.57), 1.941 (0.60), 1.959 (0.43), 2.179 (0.46), 2.196 (0.55), 2.2 10 (0.49), 2.331 (13.92), 2.348 (0.45), 2.518 (0.65), 2.523 (0.40), 3.298 (0.66), 3.308 (0.74), 3.325 (1.11), 3.519 (0.43), 3.525 (0.53), 3.532 (0.43), 3.537
(0.58), 3.544 (0.52), 3.558 (0.40), 3.597 (0.41), 3.616 (0.87), 3.634 (0.49), 3.641 (0.61), 3.663 (0.87 ), 3.678 (0.91), 3.689 (0.78), 3.705 (0.69), 4.391 (0.59), 4.404 (0.58), 5.578 (0.67), 5.597 (0.98), 5.615 (0.66), 7.032 (2.76), 7.059 (0.70), 7.065 (0.52), 7.0 76 (0.58), 7.083 (1.35), 7.088 (1.03), 7.105 (0.75), 7.112 (0.79), 7.122 (1.56), 7.128 (1.78), 7.145 (1.92), 7.149 (1.36), 8.190 (1.12), 8.207 (1.12), 8.300 (1.12), 8.319 (1.09), 8.646 (4.19). Example 98 N‐[(3R)‐1‐(4‐{[(1S)‐1‐(3,5‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide See example 97 for details. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.571 (4.85) , 1.589 (4.97), 1.825 (16.00), 1.907 (0.52), 1.926 (0.58), 1.940 (0.60), 1.957 (0.44), 2.175 (0.50), 2.1 91 (0.56), 2.206 (0.50), 2.330 (14.07), 2.518 (0.79), 2.523 (0.50), 3.311 (0.76), 3.321 (0.97), 3.551 (0.57), 3.558 (0.40), 3.565 (0.66), 3.571 (0.63), 3.583 (0.77), 3.600 (1.00), 3.618 (0.53), 3.625 (0.52), 3.6 38 (0.82), 3.654 (0.91), 3.665 (0.79), 3.680 (0.69), 4.395 (0.60), 4.409 (0.58), 5.574 (0.67), 5.592 (0.98), 5.610 (0.67), 7.026 (2.78), 7.059 (0.68), 7.065 (0.51), 7.076 (0.58), 7.082 (1.32), 7.088 (1.00), 7.1 05 (0.79), 7.111 (0.83), 7.121 (1.55), 7.126 (1.77), 7.143 (1.91), 7.148 (1.36), 8.196 (1.16), 8.214 (1.14 ), 8.297 (1.15), 8.316 (1.10), 8.646 (4.11). Example 99 N‐[(3R)‐1‐(4‐{[(1S)‐1‐(2,6‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide Example 74 (80 mg, 190 µmol) was purified by chira l HPLC to give: Example 99 (25 mg, 30%, e.e. >95%). Rt = 5.00 m in Example 100 (29 mg, 34%, e.e. >95%). Rt = 3.31 mins
Analytical Method: Instrument: Thermo Fisher UltiMate 3000; Column: YMC Cellulose SB 3µ, 100x4.6; eluent A: methyl tert‐butyl ether + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 95%A+5%B; flow: 1.4 ml/min; temperature: 25°C; UV: 280 nm Preparative Method: Instrument: PrepCon Labomatic HPLC‐3; Column: YMC Cellulose SB 10µ, 250x50; eluent A: methyl tert‐butyl ether + 0.1 vol % diethylamine; eluent B: ethanol + 0.1 vol % diethylamine; isocratic: 95%A+5%B; flow: 80 ml/min; temperature: 25°C; UV: 280 nm ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.695 (4.49) , 1.713 (4.49), 1.831 (16.00), 1.918 (0.59), 1.931 (0.60), 1.950 (0.44), 2.074 (1.34), 2.173 (0.47), 2.1 90 (0.56), 2.204 (0.51), 2.220 (0.40), 2.266 (14.03), 2.277 (0.57), 2.518 (0.72), 2.523 (0.51), 3.294 (0.69), 3.304 (0.76), 3.321 (1.07), 3.535 (0.57), 3.543 (0.40), 3.549 (0.64), 3.555 (0.58), 3.569 (0.50), 3.5 75 (0.49), 3.594 (0.96), 3.602 (0.41), 3.612 (0.54), 3.620 (0.60), 3.628 (0.83), 3.644 (0.96), 3.655 (0.80), 3.670 (0.69), 4.399 (0.59), 4.412 (0.58), 5.625 (0.63), 5.642 (0.89), 5.659 (0.60), 6.984 (1.40), 6.9 93 (0.41), 7.005 (2.91), 7.018 (0.42), 7.026 (1.65), 7.117 (2.91), 7.261 (0.71), 7.265 (0.57), 7.277 (0.48), 7.282 (1.11), 7.286 (0.49), 7.297 (0.56), 7.302 (0.63), 8.203 (1.16), 8.219 (1.13), 8.389 (1.03), 8.4 06 (0.98), 8.603 (4.05). Example 100 N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,6‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide See example 99 for details. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.693 (4.37) , 1.711 (4.42), 1.825 (16.00), 1.922 (0.56), 1.936 (0.59), 1.954 (0.42), 2.174 (0.48), 2.191 (0.57), 2.2 05 (0.50), 2.221 (0.40), 2.267 (13.53), 2.283 (0.44), 2.518 (0.55), 3.299 (0.70), 3.309 (0.74), 3.326 (0.97), 3.511 (0.43), 3.517 (0.54), 3.524 (0.43), 3.530 (0.57), 3.537 (0.52), 3.603 (0.88), 3.610 (0.41), 3.6 21 (0.50), 3.629 (0.65), 3.637 (0.82), 3.653 (0.94), 3.665 (0.78), 3.680 (0.69), 4.395 (0.57), 4.409 (0.57), 5.631 (0.61), 5.649 (0.87), 5.666 (0.58), 6.984 (1.36), 7.005 (2.87), 7.026 (1.61), 7.118 (2.89), 7.2 59 (0.68), 7.264 (0.56), 7.280 (1.07), 7.297 (0.52), 7.301 (0.58), 8.197 (1.13), 8.214 (1.11), 8.387 (1.05 ), 8.403 (1.01), 8.603 (4.11). Example 101 N‐[(3R)‐1‐(4‐{[(1R)‐1‐(2,5‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide
Example 75 (80 mg, 190 µmol) was purified by chira l HPLC to give: Example 101 (32 mg, 38%, e.e. >95%). Rt = 3.34 min Example 102 (32 mg, 38%, e.e. >95%). Rt = 4.41 mins Analytical Method: Instrument: Thermo Fisher UltiMate 3000; Column: YMC Cellulose SB 3µ, 100x4.6; eluent A: methyl tert‐butyl ether + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 95%A+5%B; flow: 1.4 ml/min; temperature: 25°C; UV: 2 80 nm Preparative Method: Instrument: PrepCon Labomatic HPLC‐3; Column: YMC Cellulose SB 10µ, 250x50; eluent A: methyl tert‐butyl ether + 0.1 vol % diethylamine; eluent B: ethanol + 0.1 vol % diethylamine; isocratic: 95%A+5%B; flow: 80 ml/min; te mperature: 25°C; UV: 280 nm 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.231 (0 .59), 1.570 (4.90), 1.588 (4.94), 1.825 (16.00), 1.93 1 (0.59), 1.945 (0.61), 1.963 (0.45), 2.184 (0.47), 2.2 00 (0.56), 2.216 (0.50), 2.305 (14.24), 2.322 (0.63), 2.327 (0.72), 2.332 (0.50), 2.518 (2.73), 2.523 (1.69), 2.665 (0.43), 2.669 (0.61), 2.673 (0.43), 3.306 (0.90), 3.316 (1.26), 3.526 (0.45), 3.532 (0.54), 3.5 45 (0.61), 3.551 (0.52), 3.565 (0.41), 3.604 (0.43), 3.623 (0.90), 3.641 (0.50), 3.648 (0.63), 3.671 (0.88), 3.687 (0.95), 3.698 (0.81), 3.713 (0.72), 4.395 (0.61), 4.410 (0.61), 5.732 (0.63), 5.750 (0.97), 5.7 68 (0.63), 7.056 (2.96), 7.094 (0.45), 7.108 (0.50), 7.116 (0.88), 7.126 (0.65), 7.136 (0.63), 7.145 (0.43), 7.216 (0.72), 7.226 (0.77), 7.239 (1.24), 7.250 (1.62), 7.257 (1.17), 7.262 (0.93), 7.266 (0.70), 7.2 73 (1.11), 7.281 (0.54), 8.194 (1.15), 8.210 (1.15), 8.309 (1.20), 8.328 (1.15), 8.642 (4.36). Example 102 N‐[(3R)‐1‐(4‐{[(1S)‐1‐(2,5‐difluorophenyl)ethy l]amino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl)pyrrolidin‐3‐yl]acetamide See example 101 for details. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.572 (5.06) , 1.589 (5.07), 1.830 (16.00), 1.929 (0.63), 1.943 (0.65), 1.960 (0.48), 2.179 (0.53), 2.195 (0.63), 2.2 10 (0.55), 2.226 (0.44), 2.304 (14.05), 2.518 (0.63),
3.320 (0.92), 3.330 (1.31), 3.552 (0.40), 3.558 (0.63), 3.572 (0.72), 3.578 (0.72), 3.587 (0.72), 3.605 (1.09), 3.623 (0.59), 3.630 (0.54), 3.645 (0.89), 3.6 60 (0.98), 3.671 (0.84), 3.687 (0.75), 4.387 (0.40), 4.401 (0.66), 4.413 (0.64), 5.726 (0.67), 5.744 (1.02), 5.762 (0.67), 7.049 (3.02), 7.091 (0.48), 7.105 (0.54), 7.113 (0.93), 7.123 (0.69), 7.133 (0.67), 7.1 43 (0.44), 7.213 (0.76), 7.225 (0.83), 7.231 (0.76), 7.237 (1.37), 7.247 (1.54), 7.254 (1.26), 7.260 (1.00), 7.271 (0.98), 7.277 (0.59), 8.204 (1.24), 8.221 (1.20), 8.308 (1.24), 8.327 (1.18), 8.643 (4.30). Example 103 3‐[(1R)‐1‐({6‐[(3R)‐3‐acetamidopyrrolidin‐1‐ yl]‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ yl}amino)ethyl]benzoic acid To a solution of Example 65 (21.2 mg, 47.3 µmol) in MeOH (2ml) was added 1M NaOH (2ml). Stirred at RT for 16h.) Reaction concentrated under reduced pressure and the residue was purified by preparative HPLC (basic method) to give the titled c ompound (13.8 mg, 64%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.571 (4.77) , 1.588 (4.82), 1.815 (16.00), 1.915 (0.63), 1.928 (0.67), 1.946 (0.46), 2.163 (0.51), 2.178 (0.65), 2.1 94 (0.57), 2.209 (0.44), 2.333 (14.27), 2.522 (0.81), 3.307 (1.33), 3.316 (1.51), 3.334 (1.99), 3.343 (2.31), 3.478 (1.15), 3.490 (1.03), 3.499 (1.08), 3.504 (1.14), 3.516 (1.07), 3.524 (0.94), 3.536 (0.74), 3.5 76 (0.61), 3.595 (1.11), 3.613 (0.71), 3.620 (0.83), 3.638 (1.17), 3.654 (1.11), 3.666 (0.95), 3.681 (0.84), 4.368 (0.42), 4.381 (0.69), 4.395 (0.69), 5.629 (0.72), 5.647 (1.04), 5.667 (0.70), 7.103 (3.09), 7.2 03 (1.12), 7.222 (2.51), 7.241 (1.46), 7.363 (1.41), 7.383 (1.14), 7.704 (1.78), 7.722 (1.65), 7.971 (2.63), 8.227 (1.41), 8.244 (1.40), 8.398 (1.29), 8.418 (1.24), 8.608 (4.76). Example 104 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(hydroxymethyl)phenyl ]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]pyrrolidin‐3‐yl}acetamide
To a solution of Example 65 (22mg, 49 µmol) in THF (3 ml) and NaBH4 (14.8 mg, 392 µmol) was added and stirred at RT for 1h. To the reaction mi xture was added MeOH (3 ml) and stirred at RT for 3h. The reaction was concentrated and the residue wa s purified by preparative HPLC (basic method) to give the titled compound (3.4 mg, 16%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.569 (5.26) , 1.587 (5.32), 1.821 (16.00), 1.902 (0.45), 1.920 (0.71), 1.933 (0.75), 1.951 (0.56), 2.173 (0.66), 2.1 89 (0.75), 2.204 (0.68), 2.221 (0.52), 2.334 (14.38), 2.669 (0.41), 3.288 (1.00), 3.298 (1.31), 3.315 (1.92), 3.492 (0.42), 3.517 (0.74), 3.530 (0.78), 3.537 (0.69), 3.550 (0.52), 3.588 (0.48), 3.606 (1.04), 3.6 24 (0.62), 3.631 (0.73), 3.647 (1.09), 3.662 (1.05), 3.674 (0.90), 3.689 (0.79), 4.374 (0.46), 4.386 (0.78), 4.400 (0.77), 4.471 (5.94), 5.611 (0.77), 5.629 (1.13), 5.647 (0.75), 7.065 (3.32), 7.149 (1.24), 7.1 67 (1.60), 7.248 (0.92), 7.267 (2.36), 7.286 (3.19), 7.289 (2.65), 7.308 (0.67), 7.402 (2.60), 8.194 (1.37), 8.211 (1.37), 8.316 (1.41), 8.336 (1.39), 8.618 (4.71). Example 105 N‐[(3R)‐1‐(4‐{[(1R)‐1‐(3‐hydroxyphenyl)ethyl]a mino}‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐yl)pyrrolid in‐ 3‐yl]acetamide In a vessel flushed with argon was added tBuBrettPhos Pd G3 (8.19 mg, 9.59 µmol), tBuBrettPhos (4.65 mg, 9.59 µmol), Cs2CO3 (43.7 mg, 134 µmol) and Example 62 (45.0 mg, 95.9 µmol). The vessel was flushed again with Argon and toluene (1.2 ml) a nd 2,2‐difluoroethan‐1‐ol (61 µl, 960 µmol) we re added. The reaction mixture was heated at 80°C for 16h. The reaction mixture was diluted with EtOAc, washed with water, filtered through a hydrophobic membran concentrated under vacuu m. The residue was purified by silica chromatography (DC M:EtOH) to give Example 105 (5 mg, 13%) and Example 106 (10 mg, 22%) 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.850 (0.66), 0.867 (1.18), 0.872 (0.87), 0.887 (1.18), 0.905 (1.63), 0.924 (0.69), 1.107 (1.28), 1.232 (1.49), 1.2 56 (0.52), 1.278 (0.76), 1.295 (0.76), 1.316 (0.49), 1.349 (1.28), 1.537 (4.69), 1.555 (4.69), 1.820 (16.0 0), 1.921 (0.59), 1.934 (0.62), 1.952 (0.42), 2.075 (0.83), 2.172 (0.45), 2.188 (0.56), 2.202 (0.49), 2.3 18 (0.49), 2.323 (1.08), 2.327 (1.56), 2.332 (1.56), 2.339 (14.30), 2.518 (4.65), 2.523 (3.16), 2.540 (0.5 2), 2.660 (0.45), 2.665 (0.97), 2.669 (1.35), 2.674 (0.94), 2.679 (0.45), 3.285 (0.76), 3.295 (0.94), 3.5 05 (0.56), 3.511 (0.62), 3.519 (0.56), 3.532 (0.59), 3.539 (0.52), 3.606 (0.87), 3.623 (0.49), 3.631 (0.59), 3.646 (0.87), 3.661 (0.94), 3.673 (0.80), 3.688
(0.69), 4.249 (0.42), 4.371 (0.42), 4.386 (0.62), 4.398 (0.59), 5.555 (0.62), 5.574 (0.90), 5.592 (0.62 ), 6.591 (1.01), 6.595 (1.01), 6.597 (0.97), 6.611 (1.04), 6.615 (1.11), 6.813 (2.12), 6.818 (1.49), 6.836 (1.21), 6.855 (1.39), 7.064 (2.88), 7.086 (1.67), 7.1 05 (2.57), 7.124 (1.25), 8.088 (2.08), 8.185 (1.21), 8.202 (1.18), 8.251 (1.25), 8.271 (1.15), 8.624 (4.23 ), 9.304 (2.74). Example 106 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(2,2‐difluoroethoxy )phenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐ yl]pyrrolidin‐3‐yl}acetamide See Example 105 for details. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (0 .69), 1.563 (4.39), 1.581 (4.42), 1.820 (16.00), 1.92 2 (0.55), 1.936 (0.57), 2.174 (0.48), 2.191 (0.53), 2.2 07 (0.46), 2.322 (1.01), 2.333 (13.97), 2.518 (2.95), 2.522 (2.01), 2.664 (0.70), 2.668 (0.95), 2.673 (0.67), 3.294 (0.67), 3.304 (0.83), 3.516 (0.42), 3.521 (0.52), 3.528 (0.42), 3.534 (0.56), 3.541 (0.49), 3.6 09 (0.84), 3.627 (0.46), 3.634 (0.57), 3.653 (0.94), 3.668 (0.88), 3.679 (0.76), 3.695 (0.67), 4.244 (0.78), 4.252 (0.81), 4.281 (1.59), 4.289 (1.55), 4.317 (0.84), 4.326 (0.74), 4.388 (0.57), 4.402 (0.56), 5.5 91 (0.60), 5.609 (0.88), 5.628 (0.60), 6.230 (0.69), 6.358 (0.62), 6.366 (1.40), 6.375 (0.64), 6.502 (0.59), 6.859 (0.80), 6.861 (0.83), 6.865 (0.84), 6.868 (0.85), 6.881 (0.95), 6.886 (1.02), 7.052 (5.41), 7.0 60 (1.59), 7.068 (1.48), 7.247 (1.57), 7.268 (2.60), 7.288 (1.17), 8.186 (1.16), 8.203 (1.13), 8.262 (1.16 ), 8.282 (1.12), 8.626 (3.97). Example 107 N‐[(3R)‐1‐(4‐{[(1R)‐1‐{3‐[(E)‐2‐ethoxyethe nyl]phenyl}ethyl]amino}‐2‐methylpyrido[3,4‐d]pyrimidin ‐6‐ yl)pyrrolidin‐3‐yl]acetamide To a solution of Example 62 (600 mg, 1.28 mmol) in dioxane (8.1 ml) was added 2‐[(E)‐2‐ ethoxyethenyl]‐4,4,5,5‐tetramethyl‐1,3,2‐dioxaborola ne (253 mg, 1.28 mmol), followed by K2CO3 (589 mg, 4.26 mmol) and Pd(PPh3)4 (123 mg, 107 µmol) and water (1.62 ml). The reaction was heated at 90°C for 16h. The reaction was concentrated and purified by silica chromatography (EtOH:DCM) to give the titled compound (480 mg, 81%) . 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.222 (4.24), 1.239 (9.16), 1.256 (4.35), 1.562 (4.62), 1.579 (4.65), 1.819 (16.00), 1.919 (0.59), 1.931 (0.65), 1. 949 (0.46), 2.171 (0.49), 2.187 (0.59), 2.202 (0.54), 2.218 (0.41), 2.323 (1.30), 2.327 (1.95), 2.331 (1.86 ), 2.341 (13.89), 2.518 (10.73), 2.523 (7.46), 2.665 (1.30), 2.669 (1.76), 2.673 (1.27), 3.286 (0.73), 3.2 96 (0.76), 3.518 (0.59), 3.531 (0.65), 3.551 (0.43), 3.586 (0.43), 3.605 (0.95), 3.623 (0.54), 3.630 (0.65), 3.647 (1.00), 3.662 (0.97), 3.673 (0.84), 3.689 (0.76), 3.846 (1.16), 3.864 (3.81), 3.881 (3.81), 3.8 99 (1.16), 4.385 (0.65), 4.400 (0.68), 5.574 (0.70), 5.593 (0.95), 5.611 (0.65), 5.794 (2.24), 5.827 (2.38), 7.055 (2.97), 7.143 (0.76), 7.148 (1.05), 7.153 (1.05), 7.169 (4.08), 7.177 (2.73), 7.188 (1.11), 7.2 02 (2.89), 7.213 (0.46), 7.314 (2.30), 8.185 (1.30), 8.202 (1.24), 8.261 (1.24), 8.280 (1.19), 8.622 (4.27 ). Table 4: Examples 108‐115 Using the method described for Example 25: Intermediate 11 was treated with the corresponding phenylethan‐1‐amines or their hydrochloride salts a nd gave the desired compounds after preparative HPLC purification (basic method) and/or optionally sil ica chromatography.
Table 5: Examples 116‐122 Using the method described for Example 25: Intermediate 12 was treated with the corresponding phenylethan‐1‐amines or their hydrochloride salts a nd gave the desired compounds after preparative HPLC purification (basic method) and/or optionally sil ica chromatography.
Table 6: Examples 123‐129 Using the method described for Example 25: Intermediate 13 was treated with the corresponding phenylethan‐1‐amines or their hydrochloride salts a nd gave the desired compounds after preparative HPLC purification (basic method) and/or optionally sil ica chromatography.
Table 7: Examples 130‐136 Using the method described for Example 25: Intermediate 14 was treated with the corresponding phenylethan‐1‐amines or their hydrochloride salts a nd gave the desired compounds after preparative HPLC purification (basic method) and/or optionally sil ica chromatography.
Table 8: Examples 137‐257 Using the method described for Example 3: Example 2 was treated with nitrogen containing nucleophile at 130°C. The desired compounds were obtainied after preparative HPLC purification (basic method) and/or optionally silica chromatography.
Example 258 methyl 4‐(2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl )‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethoxy)benzoate A mixture of Example 2 (50.0 mg, 143 µmol), methyl 4‐[2‐(piperazin‐1‐yl)ethoxy]benzoate hydrochloride (144 mg, 428 µmol) and DIPEA (150 µl , 860 µmol) in DMSO (1 ml) was heated at 130°C for 16h. The titled compound was isolated (10 mg, 20%) after preparative HPLC purification (basic method). 1H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.594 (1.73), 1.605 (4.49), 1.612 (2.38), 1.623 (4.38), 2.302 (12.90), 2.318 (0.66), 2.322 (1.18), 2.327 (1.56), 2. 332 (1.15), 2.336 (0.55), 2.401 (4.33), 2.518 (6.52), 2.523 (4.11), 2.539 (0.60), 2.665 (3.26), 2.669 (3.62), 2.674 (3.84), 2.688 (2.33), 2.725 (3.21), 2.800 (1.18), 2.813 (2.41), 2.827 (1.21), 3.582 (2.79), 3.8 13 (16.00), 4.231 (1.32), 4.245 (2.68), 4.259 (1.26), 5.750 (1.01), 5.768 (1.23), 5.786 (0.71), 7.069 (0.55), 7.076 (4.16), 7.081 (1.29), 7.094 (1.40), 7.099 (4.79), 7.237 (2.58), 7.275 (0.82), 7.295 (1.75), 7.3 14 (1.01), 7.373 (1.07), 7.445 (2.52), 7.488 (0.71), 7.504 (1.18), 7.523 (0.58), 7.632 (0.55), 7.650 (1.12), 7.667 (0.77), 7.898 (0.68), 7.905 (4.74), 7.911 (1.37), 7.923 (1.34), 7.928 (4.27), 7.935 (0.47), 7.9 45 (0.60), 8.426 (1.10), 8.445 (1.04), 8.659 (4.16). Example 259 4‐(2‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐ fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimid in‐ 6‐yl]piperazin‐1‐yl}ethoxy)benzoicacid To a solution of Example 258 (8.20 mg, 13.8 µmol) in MeOH (2ml) was added 1M NaOH (2ml), additional MeOH (1ml) needed for homogeneous solution. Stirred at RT for 16h. Reaction was concentrated under reduced pressure to remove MeOH. Dissolved in DMSO:water (1:1) The titled compound was isolated (3.4 mg, 40%) after preparative HPLC purification (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.605 (5.44) , 1.623 (5.45), 2.301 (16.00), 2.322 (0.47), 2.326 (0.52), 2.518 (1.82), 2.522 (1.17), 2.668 (4.18), 2.6 77 (3.07), 2.784 (1.43), 2.799 (2.90), 2.813 (1.53), 3.383 (2.18), 3.578 (4.14), 4.193 (1.62), 4.207 (3.21), 4.221 (1.57), 5.751 (0.87), 5.769 (1.34), 5.787 (0.85), 6.985 (3.99), 7.008 (4.09), 7.101 (1.25), 7.2 37 (2.74), 7.272 (0.99), 7.291 (2.13), 7.310 (1.23), 7.372 (1.10), 7.467 (3.18), 7.483 (0.87), 7.499 (1.31), 7.517 (0.64), 7.651 (0.70), 7.669 (1.28), 7.687 (0.64), 7.858 (5.09), 7.862 (1.65), 7.875 (1.64), 7.8 79 (4.58), 8.513 (1.21), 8.532 (1.14), 8.656 (5.55). Example 260 6‐(methanesulfonyl)‐2‐methyl‐N‐{(1R)‐1‐[3‐(t rifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐ amine A mixture of Example 10 (50.0 mg, 143 µmol) and s odium methanesulfinate (72.9 mg, 714 µmol) in DMSO (1 ml) was heated at 130°C for 16h. The titled compound was isolated (14 mg, 23%) after preparative HPLC purification (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.625 (1.19), 1.643 (1.20), 2.482 (4.12), 2.518 (0.67), 2.523 (0.44), 3.313 (4.58), 3.331 (16.00), 5.758 (4.13), 7. 607 (0.44), 7.846 (0.49), 9.089 (1.10), 9.105 (0.97). Example 261
6‐[(3R)‐3‐aminopyrrolidin‐1‐yl]‐N‐{(1R)‐1 ‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride s alt x.HCl To an ice‐cooled solution of Example 250 (960 mg, 1.86 mmol) in dioxane (4.1 ml) was added HCL in dioxane (4.1 ml, 4.0 M, 16 mmol) and stirred for 3 h. The reaction was concentrated under reduced pressure to give the titled compound (904 mg) which was used without further purification. 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.748 (2.93), 1.765 (2.93), 2.374 (0.53), 2.392 (0.44), 2.518 (0.95), 2.523 (0.71), 2.539 (11.48), 3.161 (8.17), 3. 561 (0.42), 3.599 (0.49), 3.678 (0.48), 3.710 (0.67), 3.750 (0.58), 3.770 (0.48), 3.788 (0.74), 3.803 (0.82), 3.818 (0.46), 3.834 (0.46), 3.983 (0.54), 5.758 (16.00), 5.978 (0.63), 5.996 (0.94), 6.013 (0.59), 7. 105 (1.06), 7.241 (2.23), 7.329 (0.50), 7.348 (1.07), 7.376 (1.06), 7.542 (0.51), 7.559 (0.84), 7.900 (0.72), 7.942 (0.49), 7.959 (0.61), 8.534 (0.83), 8.864 (2.51). Example 262 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}cyclopropanecarboxa mide To a solution of example 261 (50.0 mg, 110 µmol) and cyclopropanecarboxylic acid (18 µl, 220 µmol) in DMF (830µl) was added DIPEA (96 µl, 550 µmol) and propylphosphonic anhydride solution (T3P) in DMF (130 µl, 50 % purity, 220 µmol). Tot he reac tion was added a few drops of water and the the titled compound (29 mg, 52%) was isolated after prep arative HPLC (basc method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.638 (1.36), 0.643 (1.84), 0.654 (1.40), 0.656 (1.63), 0.663 (1.86), 0.669 (1.89), 0.686 (2.52), 0.699 (1.20), 0.7 12 (0.46), 1.107 (0.61), 1.231 (0.74), 1.551 (0.64), 1.558 (0.74), 1.570 (1.12), 1.582 (0.67), 1.589 (0.76), 1.603 (5.13), 1.621 (5.06), 1.959 (0.59), 1.973 (0.64), 1.991 (0.46), 2.197 (0.48), 2.214 (0.61), 2.2 30 (0.54), 2.245 (0.41), 2.290 (16.00), 2.322 (0.56), 2.326 (0.72), 2.332 (0.51), 2.518 (3.04), 2.522 (1.87), 2.539 (0.41), 2.664 (0.49), 2.669 (0.69), 2.673 (0.51), 3.295 (0.72), 3.306 (0.89), 3.536 (0.49), 3.5 42 (0.58), 3.549 (0.48), 3.556 (0.62), 3.562 (0.56),
3.635 (0.41), 3.653 (0.90), 3.674 (1.17), 3.689 (1.07), 3.700 (1.00), 3.716 (0.76), 4.410 (0.39), 4.425 (0.66), 4.437 (0.66), 5.762 (0.77), 5.780 (1.18), 5.7 98 (0.77), 7.088 (3.16), 7.100 (1.28), 7.237 (2.53), 7.275 (0.87), 7.294 (1.92), 7.313 (1.12), 7.372 (1.02), 7.482 (0.66), 7.500 (1.09), 7.518 (0.54), 7.627 (0.59), 7.645 (1.09), 7.663 (0.54), 8.402 (1.35), 8.4 20 (2.61), 8.436 (1.41), 8.636 (4.72). Example 263 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐2,2‐difluoroac etamide Using the method described for Example 262: Example 261 (60 mg, 132 µmol) and difluoroacetic acid (17 µl, 260 µmol) gave the titled compound (39 mg , 56%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (8.91), 1.605 (5.01), 1.622 (5.05), 2.056 (0.61), 2.069 (0.66), 2.088 (0.47), 2.253 (0.53), 2.270 (0.70), 2.2 93 (16.00), 2.322 (0.54), 2.327 (0.55), 2.518 (2.15), 2.523 (1.31), 2.539 (0.64), 2.669 (0.50), 3.401 (0.72), 3.411 (0.76), 3.428 (0.82), 3.438 (0.82), 3.543 (0.50), 3.549 (0.59), 3.562 (0.66), 3.568 (0.59), 3.5 82 (0.42), 3.628 (0.42), 3.646 (0.92), 3.663 (0.53), 3.672 (0.65), 3.725 (0.82), 3.741 (1.01), 3.752 (0.87), 3.768 (0.77), 4.190 (0.57), 4.507 (0.65), 4.520 (0.65), 5.763 (0.78), 5.781 (1.20), 5.800 (0.77), 6.0 79 (1.61), 6.214 (3.89), 6.348 (1.41), 7.100 (4.19), 7.237 (2.49), 7.276 (0.88), 7.295 (1.92), 7.314 (1.11), 7.372 (1.02), 7.484 (0.66), 7.501 (1.13), 7.520 (0.56), 7.629 (0.61), 7.646 (1.10), 7.664 (0.55), 8.3 98 (1.32), 8.416 (1.27), 8.644 (4.72), 9.160 (1.21), 9.178 (1.19). Example 264 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐2‐methoxyaceta mide Using the method described for Example 262: Example 261 (50 mg, 110 µmol) and methoxyacetic acid (19.9 mg, 221 µmol) gave the titled compound (35 mg, 61%) after preparative HPLC (basic method).
¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (0.99), 1.602 (2.93), 1.620 (2.94), 2.289 (9.20), 2.327 (0.42), 2.518 (1.78), 2.522 (1.10), 2.669 (0.42), 3.3 02 (16.00), 3.364 (0.58), 3.377 (0.52), 3.524 (0.43), 3.647 (0.41), 3.698 (0.49), 3.714 (0.58), 3.724 (0.52), 3.741 (0.45), 3.829 (4.30), 4.495 (0.43), 4.509 (0.42), 5.761 (0.45), 5.779 (0.69), 5.798 (0.45), 7.0 76 (1.85), 7.100 (0.69), 7.237 (1.45), 7.274 (0.50), 7.293 (1.13), 7.312 (0.65), 7.373 (0.60), 7.500 (0.64), 7.647 (0.64), 8.119 (0.80), 8.137 (0.79), 8.390 (0.76), 8.409 (0.73), 8.630 (2.77). Example 265 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}oxetane‐3‐carbo xamide Using the method described for Example 262: Example 261 (50 mg, 110 µmol) and oxetane‐3‐ carboxylic acid (22.5 mg, 221 µmol) gave the titled compound (31 mg, 53%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (1.39), 1.232 (0.90), 1.601 (5.07), 1.619 (5.07), 1.940 (0.62), 1.953 (0.64), 1.971 (0.45), 2.202 (0.51), 2.2 17 (0.62), 2.233 (0.56), 2.249 (0.41), 2.288 (16.00), 2.322 (0.66), 2.326 (0.90), 2.332 (0.64), 2.518 (3.21), 2.522 (1.90), 2.539 (0.43), 2.664 (0.60), 2.669 (0.83), 2.673 (0.62), 3.304 (0.88), 3.315 (1.16), 3.5 39 (0.60), 3.552 (0.66), 3.559 (0.60), 3.572 (0.49), 3.582 (0.47), 3.600 (1.01), 3.618 (0.53), 3.626 (0.58), 3.694 (0.79), 3.710 (1.03), 3.722 (1.50), 3.725 (1.26), 3.743 (1.71), 3.763 (1.01), 4.459 (0.64), 4.4 72 (0.64), 4.593 (5.37), 4.611 (7.94), 4.613 (4.98), 4.628 (3.32), 4.632 (3.19), 4.646 (0.41), 5.760 (0.77), 5.778 (1.22), 5.796 (0.77), 7.081 (3.14), 7.100 (1.22), 7.236 (2.57), 7.276 (0.90), 7.295 (1.95), 7.3 14 (1.11), 7.372 (1.05), 7.483 (0.66), 7.501 (1.11), 7.519 (0.53), 7.627 (0.60), 7.645 (1.09), 7.663 (0.53), 8.245 (1.35), 8.262 (1.33), 8.395 (1.30), 8.414 (1.24), 8.630 (4.77). Example 266 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}‐1‐methylazetid ine‐3‐carboxamide
Using the method described for Example 262: Example 261 (50 mg, 110 µmol) and 1‐ methylazetidine‐3‐carboxylic acid (25.4 mg, 221 µmol) gave the titled compound (12.5 mg, 21%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.103 (1.38), 1.107 (1.89), 1.224 (1.03), 1.230 (1.16), 1.601 (4.38), 1.619 (4.45), 1.826 (0.75), 1.926 (0.50), 1.9 40 (0.52), 2.160 (11.15), 2.183 (0.67), 2.198 (0.63), 2.214 (0.52), 2.288 (16.00), 2.322 (0.84), 2.327 (0.7 5), 2.332 (0.55), 2.518 (2.88), 2.523 (1.76), 2.539 (0.56), 2.665 (0.50), 2.669 (0.72), 2.673 (0.52), 3.0 38 (0.50), 3.055 (1.15), 3.072 (1.99), 3.077 (1.47), 3.087 (1.35), 3.104 (0.75), 3.122 (0.59), 3.268 (1.36), 3.278 (1.60), 3.294 (2.06), 3.346 (7.98), 3.460 (0.74), 3.496 (0.67), 3.510 (0.65), 3.516 (0.69), 3.5 23 (0.75), 3.531 (0.65), 3.537 (0.76), 3.542 (0.73), 3.556 (0.55), 3.592 (0.48), 3.610 (0.88), 3.618 (0.75), 3.628 (0.80), 3.636 (0.85), 3.644 (0.65), 3.679 (0.81), 3.694 (0.98), 3.706 (0.96), 3.721 (0.80), 4.4 15 (0.53), 4.428 (0.52), 5.759 (0.80), 5.779 (1.19), 5.796 (0.79), 7.064 (0.84), 7.074 (2.45), 7.100 (1.39), 7.236 (2.80), 7.275 (0.82), 7.295 (1.76), 7.314 (1.04), 7.372 (1.20), 7.483 (0.79), 7.500 (1.33), 7.5 18 (0.67), 7.628 (0.64), 7.646 (1.14), 7.665 (0.59), 8.161 (0.90), 8.178 (0.90), 8.398 (1.20), 8.416 (1.16 ), 8.629 (3.85). Example 267 methyl {(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}carbamate To a mixture of Example 261 (50 mg, 110 µmol), tr iethylamine (77 µl, 550 µmol), DMAP (0.3 mg) in DCE (830 µl) was stirred at RT for 16h. The react ion mixture was added water, extracted with DCM, washed with sat. NaCl. The organics were filtered th rough a hydrophobic filter and concentrated. The titled compound (13 mg, 24%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (1.81), 1.231 (0.85), 1.603 (5.33), 1.621 (5.42), 1.952 (0.54), 1.966 (0.61), 1.987 (0.52), 2.187 (0.54), 2.2 05 (0.67), 2.219 (0.63), 2.237 (0.46), 2.288 (16.00), 2.322 (0.81), 2.327 (1.09), 2.331 (0.83), 2.518 (7.66), 2.523 (5.33), 2.539 (1.65), 2.665 (0.74), 2.669 (1.09), 2.673 (0.81), 3.512 (0.72), 3.552 (6.68), 3.6 06 (0.59), 3.624 (0.96), 3.642 (0.63), 3.648 (0.70), 3.675 (0.83), 3.691 (0.96), 3.702 (0.85), 3.718 (0.74), 4.203 (0.46), 4.219 (0.76), 4.231 (0.72), 4.245 (0.44), 5.759 (1.28), 5.778 (1.26), 5.796 (0.83), 7.0 68 (3.35), 7.101 (1.28), 7.237 (2.61), 7.274 (0.98), 7.293 (2.11), 7.312 (1.22), 7.373 (1.11), 7.483 (0.72), 7.500 (1.22), 7.517 (0.65), 7.570 (0.83), 7.586 (0.83), 7.628 (0.67), 7.646 (1.20), 7.664 (0.61), 8.3 93 (1.31), 8.411 (1.31), 8.625 (5.01). Example 268 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]pyrrolidin‐3‐yl}methanesulfonamide Using the method described for Example 262: Example 261 (50 mg, 110 µmol) and methanesulfonyl chloride (17 µl, 220 µmol) gave the titled compound (27 mg, 46%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (2.65), 1.231 (0.47), 1.609 (4.33), 1.626 (4.34), 1.999 (0.48), 2.012 (0.54), 2.030 (0.47), 2.290 (14.09), 2. 309 (0.63), 2.322 (0.73), 2.326 (0.89), 2.332 (0.54), 2.518 (2.21), 2.522 (1.33), 2.669 (0.55), 3.004 (16.0 0), 3.318 (0.48), 3.363 (0.71), 3.377 (0.71), 3.389 (0.76), 3.403 (0.76), 3.459 (0.62), 3.467 (0.48), 3.4 77 (0.41), 3.485 (0.77), 3.625 (0.54), 3.632 (0.44), 3.639 (0.47), 3.646 (0.45), 3.761 (0.66), 3.777 (0.84), 3.787 (0.71), 3.803 (0.66), 4.105 (0.62), 4.120 (0.60), 5.763 (0.67), 5.781 (1.03), 5.799 (0.66), 7.0 84 (2.75), 7.101 (1.07), 7.237 (2.13), 7.274 (0.76), 7.293 (1.66), 7.312 (0.96), 7.373 (0.89), 7.483 (0.60), 7.499 (1.99), 7.514 (1.48), 7.630 (0.51), 7.648 (0.93), 7.667 (0.47), 8.407 (1.13), 8.426 (1.07), 8.6 36 (4.14). Example 269 N‐{(3R)‐1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2 fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]p yrrolidin‐3‐yl}cyclopropanesulfonamide Using the method described for Example 262: Example 261 (50 mg, 110 µmol) and cyclopropanesulfonyl chloride (22 µl, 220 µmol) gave the titled compound (31 mg, 51%) after preparative HPLC (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.946 (0.72), 0.952 (1.27), 0.960 (2.24), 0.963 (2.18), 0.972 (2.54), 0.977 (3.04), 0.987 (1.21), 0.990 (1.23), 0.9 93 (1.39), 0.997 (1.95), 1.016 (0.43), 1.107 (3.52), 1.231 (0.43), 1.608 (5.02), 1.626 (5.03), 2.010 (0.44), 2.028 (0.54), 2.041 (0.62), 2.059 (0.55), 2.290 (16.00), 2.322 (1.03), 2.327 (0.84), 2.332 (0.80), 2. 518 (2.56), 2.523 (1.58), 2.539 (0.67), 2.635 (0.71), 2.639 (0.47), 2.651 (1.16), 2.659 (0.67), 2.665 (1.11), 2.669 (1.00), 3.384 (0.78), 3.397 (0.80), 3.410 (0.86), 3.423 (0.84), 3.464 (0.71), 3.471 (0.56), 3.4 82 (0.47), 3.488 (0.88), 3.634 (0.62), 3.639 (0.51), 3.647 (0.54), 3.653 (0.51), 3.772 (0.74), 3.789 (0.96), 3.799 (0.82), 3.816 (0.74), 4.115 (0.43), 4.131 (0.76), 4.147 (0.74), 5.763 (0.78), 5.780 (1.21), 5.7 98 (0.76), 7.085 (3.17), 7.101 (1.25), 7.237 (2.52), 7.273 (0.87), 7.293 (1.93), 7.312 (1.12), 7.373 (1.04), 7.483 (0.64), 7.501 (1.11), 7.518 (0.56), 7.539 (1.83), 7.556 (1.77), 7.630 (0.60), 7.648 (1.10), 7.6 65 (0.55), 8.401 (1.30), 8.420 (1.25), 8.637 (4.81). Table 9: Examples 270‐278 General method: To a solution of the carboxylic acid (230 µmol) in DMF (1 ml) was added HATU (230 µmol) and stirred for 15 mins, then DIPEA (766 µmol) and Example 148 (75mg, 153 µmol) were added. The reaction was stirred at RT. The compounds in Table 9 were then purified by preparative HPLC (basic method) and/or silica chromatograpy.
Example 279 10‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluo rophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin 6‐ yl]piperazin‐1‐yl}‐10‐oxodecanoic acid To a solution of Example 278 (195 mg, 317 µmol) in MeOH (5.5) and THF (1.5 ml) under Argon was added LiOH (1M in water, 1.9 ml). The reaction was stirred at RT for 16h and then neutralized by the addition of 2M HCl and concentrated. The residue was purified by silica chromatography to give the title compound (185 mg, 92%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.033 (1.69), 1.051 (3.72), 1.068 (1.55), 1.269 (4.16), 1.465 (0.73), 1.482 (1.16), 1.498 (1.22), 1.515 (1.03), 1.6 88 (1.77), 1.704 (1.81), 1.907 (0.63), 2.167 (1.77), 2.185 (3.41), 2.204 (1.61), 2.351 (1.10), 2.370 (1.69), 2.388 (0.99), 2.444 (1.45), 2.518 (4.86), 2.523 (3.25), 2.539 (16.00), 3.162 (0.69), 3.170 (0.76), 3. 409 (0.57), 3.427 (1.00), 3.444 (0.96), 3.622 (4.67), 3.705 (0.84), 7.104 (0.81), 7.240 (1.75), 7.309 (0.49), 7.328 (1.04), 7.347 (0.59), 7.376 (0.75), 7.520 (0.41), 7.537 (0.69), 8.777 (0.80). Example 280
4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorop henyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6 yl]‐ N,N‐dimethylpiperazine‐1‐carboxamide To a solution of dimethylcarbamyl chloride (25.7 mg, 239 µmol) in anhydrous THF (1 ml) was added triethylamine (67 µl, 480 µmol) followed by the sl ow addition of Example 148 (78mg, 159 µmol). The reaction was stirred at RT for 2h and then a few drops of water were added‐ The titled compound (36 mg, 45%) was isolated after preparative HPLC (basic method). ¹H‐NMR (400 MHz, CHLOROFORM‐d) δ [ppm]: 1.041 ( 0.50), 1.262 (0.58), 1.283 (0.79), 1.727 (1.41), 1.743 (1.40), 2.540 (6.00), 2.907 (16.00), 3.427 (1.0 4), 3.439 (1.43), 3.444 (1.12), 3.452 (1.42), 3.618 (1.24), 3.634 (1.26), 3.642 (0.81), 6.505 (0.42), 6.7 92 (0.45), 6.929 (0.91), 7.067 (0.43), 7.219 (0.75), 7.238 (0.43), 7.518 (0.42), 8.893 (1.48). Example 281 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐6‐[4‐(methanesulfonyl)piperazin‐1‐yl]‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine To a solution of Example 148 (88 mg, 180 µmol) in DCM (1.1. ml) was added triethylamine (75 µl, 540 µmol) followed by the slow addition of methanesulfon yl chloride (30.9 mg, 270 µmol). The reaction was stirred at RT for 2h and then a few drops of water were added‐ The titled compound (65 mg, 72%) was isolated after preparative HPLC (basic metho d). ¹H‐NMR (400 MHz, CHLOROFORM‐d) δ [ppm]: 1.040 ( 0.74), 1.194 (0.58), 1.261 (0.59), 1.282 (5.18), 1.299 (0.55), 1.730 (5.23), 1.748 (5.24), 2.543 (14.9 2), 2.844 (16.00), 3.403 (3.13), 3.415 (4.63), 3.427 (3.69), 3.730 (3.57), 3.743 (4.20), 3.755 (3.07), 5.7 73 (0.75), 5.791 (1.23), 5.809 (0.91), 5.878 (0.75), 5.895 (0.56), 6.552 (3.05), 6.783 (1.07), 6.920 (2.15), 7.058 (1.02), 7.196 (0.86), 7.216 (1.89), 7.235 (1.09), 7.497 (0.68), 7.514 (1.18), 7.533 (1.19), 7.5 53 (1.20), 7.571 (0.59), 8.893 (4.07). Example 282
2‐amino‐1‐{4‐[4‐({(1R)‐1‐[3‐(difluorometh yl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4 d]pyrimidin‐6‐yl]piperazin‐1‐yl}ethan‐1‐one To a solution of Example 148 (1g, 2.04 mmol) and N ‐Boc Glycine (537 mg, 3.07 mmol) in DMF (20 ml) under argon was added DIPEA (1.78 ml, 10.2 mmol) an d HATU (1.165 g, 3.07 mmol) and stirred at RT for 16h. The reaction was diluted with EtOAc, washed with water, sat. NaCl, dried over Na2SO4, filtered and concentrated under reduced pressure. The Boc‐protected product was purified by silica chromatography (DCM:EtOH). The Boc‐protected product was treated with 4M HCl in dioxane, concentrated and a portion was purified by preparative HPLC (basic method) to give the titled compound. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.612 (5.02) , 1.630 (5.00), 2.309 (16.00), 2.322 (0.79), 2.327 (0.76), 2.332 (0.56), 2.518 (2.20), 2.523 (1.50), 2.6 65 (0.46), 2.669 (0.65), 2.673 (0.47), 3.411 (7.92), 3.563 (2.70), 3.603 (1.56), 3.613 (1.53), 3.652 (1.25), 3.663 (1.36), 5.755 (0.76), 5.773 (1.17), 5.791 (0.76), 7.102 (1.14), 7.238 (2.50), 7.278 (0.87), 7.2 97 (1.88), 7.317 (1.08), 7.374 (1.01), 7.481 (2.98), 7.507 (1.10), 7.525 (0.53), 7.636 (0.59), 7.654 (1.07), 7.672 (0.54), 8.456 (1.22), 8.475 (1.16), 8.684 (4.80). Example 283 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐ yl]piperazin‐1‐yl}‐2‐(methylamino)ethan‐1‐one To a solution of Example 148 (1g, 2.04 mmol) and N ‐Boc Sarcosine (580 mg, 3.07 mmol) in DMF (20 ml) under argon was added DIPEA (1.78 ml, 10.2 mmol ) and HATU (1.165 g, 3.07 mmol) and stirred at RT for 16h. The reaction was diluted with EtOAc, wa shed with water, sat. NaCl, dried over Na2SO4, filtered and concentrated under reduced pressure. The Boc‐protected product was purified by silica chromatography (DCM:EtOH).
The Boc‐protected product was treated with 4M HCl in dioxane, concentrated and a portion was purified by preparative HPLC (basic method) to give the titled compound. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.612 (3.51) , 1.630 (3.49), 2.296 (16.00), 2.310 (11.42), 2.322 (0.55), 2.327 (0.51), 2.518 (1.24), 2.523 (0.83), 2.6 69 (0.41), 3.385 (5.64), 3.576 (0.99), 3.608 (3.19), 3.645 (1.01), 3.658 (1.07), 5.755 (0.55), 5.773 (0.84), 5.791 (0.53), 7.102 (0.83), 7.238 (1.78), 7.278 (0.63), 7.297 (1.33), 7.317 (0.77), 7.374 (0.73), 7.4 82 (2.16), 7.507 (0.78), 7.636 (0.42), 7.655 (0.75), 8.456 (0.87), 8.474 (0.83), 8.684 (3.40). Example 284 3‐amino‐1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl) ‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐ d]pyrimidin‐6‐yl]piperazin‐1‐yl}propan‐1‐one To a solution of Example 148 (1g, 2.04 mmol) and N ‐Boc ß‐Alanine (580 mg, 3.07 mmol) in DMF (20 ml) under argon was added DIPEA (1.78 ml, 10.2 mmol ) and HATU (1.165 g, 3.07 mmol) and stirred at RT for 16h. The reaction was diluted with EtOAc, wa shed with water, sat. NaCl, dried over Na2SO4, filtered and concentrated under reduced pressure. The Boc‐protected product was purified by silica chromatography (DCM:EtOH). The Boc‐protected product was treated with 4M HCl in dioxane, concentrated and a portion was purified by preparative HPLC (basic method) to give the titled compound. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.613 (5.18), 1.631 (5.16), 1.751 (0.48), 2.300 (1.31), 2.310 (16.00), 2.322 (1.32), 2.327 (1.51), 2.332 (1.09), 2. 336 (0.49), 2.457 (1.81), 2.473 (4.64), 2.518 (4.52), 2.523 (3.04), 2.660 (0.44), 2.665 (0.96), 2.669 (1.39), 2.673 (0.97), 2.678 (0.42), 2.757 (2.15), 2.773 (4.15), 2.789 (1.71), 3.547 (1.41), 3.630 (7.37), 3.6 52 (2.07), 5.756 (0.82), 5.774 (1.24), 5.792 (0.78), 7.103 (1.26), 7.238 (2.67), 7.279 (0.93), 7.298 (1.98), 7.317 (1.14), 7.374 (1.11), 7.483 (3.14), 7.508 (1.17), 7.525 (0.56), 7.637 (0.64), 7.654 (1.13), 7.6 72 (0.58), 8.454 (1.25), 8.472 (1.18), 8.684 (4.93). Example 285 1‐{4‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluor ophenyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐ 6‐ yl]piperazin‐1‐yl}‐3‐(methylamino)propan‐1‐one To a solution of Example 148 (1g, 2.04 mmol) and N‐(tert‐butoxycarbonyl)‐N‐methyl‐beta‐alani ne (623 mg, 3.07 mmol) in DMF (20 ml) under argon was added DIPEA (1.78 ml, 10.2 mmol) and HATU (1.165 g, 3.07 mmol) and stirred at RT for 16h. The reaction was diluted with EtOAc, washed with water, sat. NaCl, dried over Na2SO4, filtered and concentrated under reduced pressure. The Boc‐ protected product was purified by silica chromatograph y (DCM:EtOH). The Boc‐protected product was treated with 4M HCl in dioxane, concentrated and a portion was purified by preparative HPLC (basic method) to give the titled compound. ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.230 (0.72), 1.614 (5.39), 1.631 (5.29), 1.751 (0.92), 1.897 (0.52), 2.311 (16.00), 2.330 (14.82), 2.518 (4.60), 2 .523 (2.81), 2.540 (0.68), 2.560 (1.21), 2.577 (2.64) , 2.594 (1.64), 2.665 (0.88), 2.669 (1.14), 2.673 (0.83), 2.747 (1.79), 2.763 (2.96), 2.780 (1.22), 3.412 (0.85), 3.424 (0.69), 3.480 (0.43), 3.552 (1.81), 3.6 34 (8.43), 5.759 (3.01), 5.774 (1.33), 5.792 (0.84), 7.103 (1.28), 7.239 (2.76), 7.278 (1.03), 7.298 (2.12), 7.317 (1.21), 7.375 (1.14), 7.488 (3.84), 7.508 (1.39), 7.526 (0.65), 7.638 (0.73), 7.656 (1.28), 7.6 74 (0.63), 8.462 (1.37), 8.479 (1.28), 8.686 (5.28). Table 10: Examples 286‐289 Using the method described for Example 20: Example 1 0 was treated with the corresponding amines or their hydrochloride salts and gave the desired compounds after preparative HPLC purification (basic method) and/or silica chromatography.
Example 290 6‐fluoro‐2‐methyl‐N‐{(1R)‐1‐[2‐methyl‐3‐ (trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4 amine Using the method described for Example 1 using 6‐fluoro‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐ol (735 mg, 4.1 mmol) and (1R)‐1‐[2‐methyl‐3‐(trifluoromethyl)phenyl]ethan 1‐amine (1.00 g, 4.92 mmol) gave the titled compound (919 mg, 58%) after silica chromatography (Hexane:EtOAc). 1H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.559 (5.53), 1.576 (5.57), 2.382 (16.00), 2.518 (2.36), 2.522 (1.58), 2.616 (6.32), 5.677 (0.83), 5.694 (1.28), 5.7 12 (0.83), 7.341 (0.67), 7.360 (1.52), 7.380 (0.89), 7.542 (1.63), 7.560 (1.32), 7.750 (1.49), 7.769 (1.34), 8.138 (2.47), 8.141 (2.47), 8.715 (4.00), 8.883 (1.22), 8.900 (1.19). Table 11: Examples 291‐295 Using the method described for Example 25: Example 1 0 was treated with the corresponding amines or their hydrochloride salts and gave the desired compounds after preparative HPLC purification (basic method) and/or silica chromatography.
Example 296 6‐fluoro‐2,8‐dimethyl‐N‐{(1R)‐1‐[3‐(trifluor omethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine To a solution of Example 10 (250 mg, 714 µmol) in DMSO (5 ml) was added DBU (213 µl, 1.4 mmol) and nitromethane (193 µl, 3.6 mmol) and stirred for 4 days at RT. The reaction was diluted with water and the solid collected by filtration and wash ed with water. The solid was dried to give the tit le compound (260 mg, 95%). 1H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.153 (0.82), 1.171 (1.59), 1.189 (0.82), 1.603 (5.61), 1.621 (5.61), 1.986 (3.24), 2.429 (16.00), 2.518 (1.22), 2. 523 (0.74), 2.539 (1.06), 2.724 (11.33), 4.016 (0.71) , 4.034 (0.70), 5.592 (0.75), 5.609 (1.14), 5.628 (0.74), 5.758 (2.79), 7.544 (0.46), 7.563 (1.48), 7.582 (1.74), 7.591 (1.88), 7.611 (0.55), 7.740 (1.27), 7.7 58 (1.00), 7.818 (2.04), 7.889 (2.05), 7.892 (2.07), 8.639 (1.19), 8.658 (1.16). Table 12: Examples 297‐300 Using the method described for Example 25: Example 296 was treated with the corresponding amines or their hydrochloride salts and gave the desired compounds after preparative HPLC purification (basic method) and/or silica chromatograph y.
Example 301 6‐fluoro‐2,8‐dimethyl‐N‐{(1R)‐1‐[2‐methyl‐ 3‐(trifluoromethyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin 4‐ amine To a solution of Example 290 (250 mg, 868 µmol) in DMSO (4.8 ml) was added DBU (205 µl, 1.4 mmol) and nitromethane (186 µl, 3.4 mmol) and stirred for 4 days at RT. The reaction was diluted with water and the solid collected by filtration and washed with water. The solid was dried to give the title compound (243 mg, 89%). 1H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.550 (5.96), 1.567 (6.02), 1.987 (0.41), 2.327 (0.61), 2.389 (16.00), 2.539 (4.42), 2.615 (7.78), 2.669 (0.66), 2. 708 (12.68), 5.665 (0.92), 5.683 (1.45), 5.700 (0.92) , 7.333 (0.80), 7.353 (1.78), 7.372 (1.06), 7.535 (1.93), 7.555 (1.60), 7.739 (1.77), 7.759 (1.61), 7.932 (2.66), 8.760 (1.45), 8.777 (1.42). Table 13: Examples 302‐307 Using the method described for Example 25: Example 301 was treated with the corresponding amines or their hydrochloride salts and gave the desired compounds after preparative HPLC purification (basic method) and/or silica chromatograph y.
Example 308 6‐chloro‐2‐methyl‐N‐{(1R)‐1‐[3‐(trifluoromet hyl)phenyl]ethyl}pyrido[3,4‐d]pyrimidin‐4‐amine To a solution of Intermediate 15 (2.00 g, 9.71 mmol ) in DMF (40 ml) were added triethylamine (4.7 ml, 34 mmol), 4‐(dimethylamino)pyridine (1 crystal) and 2,4,6‐tri(propan‐2‐yl)benzene‐1‐sulfonyl chloride (3.24 g, 10.7 mmol) at RT. The reaction mi xture was stirred at RT for 1 hour. Then (1R)‐1 ‐[3‐ (trifluoromethyl)phenyl]ethan‐1‐amine hydrochloride (2.66 g, 11.7 mmol) was added and stirred at room temperature for 16 hoursThe reaction mxiture was diluted with water and extracted with EtOAc. The organic phase was washed with water and brine, dried over anhydrous sodium sulfate and concentrated to give a residue. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate to give the titled c ompound (3.2 g, 84%). 1H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.605 (5.96), 1.622 (5.97), 2.423 (16.00), 2.518 (1.39), 2.523 (0.89), 5.589 (0.71), 5.607 (1.08), 5.625 (0.70), 7.5 48 (0.50), 7.567 (1.59), 7.586 (1.75), 7.597 (1.94), 7.617 (0.61), 7.749 (1.40), 7.767 (1.11), 7.825 (2.30 ), 8.481 (4.08), 8.831 (4.99), 8.849 (1.11). Table 14: Examples 309‐314 Using the general method: To a solution of Example 308 (100 mg, 263 µmol) in tetrahydrofuran (1.9 ml) was added the boronic acid or pinacol borate es ter (1.2eq), potassium phosphate (2 M in water, 2 eq) and methanesulfonato(2‐dicyclohexylphosphino‐2',4',6'‐tri i‐propyl‐1,1'‐biphenyl)(2'‐amino‐ 1,1'‐biphenyl‐2‐yl)palladium(II) (0.1 eq) at RT. The reaction mixture was stirred at 70 °C for 16 hours under a nitrogen atmosphere. The reaction was diluted with water and extracted with E tOAc. The desired compounds were isolated after preparative HPLC purification (basic method) and/or silica chromatography.
Table 15: Examples 315‐318 Following the method described here for Example 315, the Examples in Table 14 were used to prepare their corresponding analogs in Table 15. Example 315: To a solution of Example 311 (180 mg, 445 µmol) in MeOH (4 ml) was added palladium on activated charcoal (10%, 0.1 eq). The reaction ve ssel was flushed with hydrogen and stirred for 4h at RT. The reaction was filtered through Celite, and the filtrate was concentrated. The desired compounds were isolated after preparative HPLC purification (acidic or basic method) and/or by silica chromatography.
Example 319 methyl 2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl) phenyl]ethyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxylate
To a solution of Example 308 (3.00 g, 8.18 mmol), triethylamine (2.3 ml, 16 mmol) in MeOH (60 ml) was added [1,1'‐bis(diphenylphosphino)ferrocene]dichloropalladium(ii ) (598 mg, 818 µmol) at RT. The reaction mixture was stirred at 80 °C for 18 hours under carbon monoxide atmosphere (50 psi). The reaction mixture was filtered and the filtrate w as purified by silica gel column chromatography (petroleum ether: EtOAc) to give the title compound (820 mg, 24%). Example 320 2‐methyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phenyl]e thyl}amino)pyrido[3,4‐d]pyrimidine‐6‐carboxamide Ammonia gas was bubling to ethanol to give a colorl ess solution at ‐65 °C. To the solution was adde d Example 319 (100 mg, 251 µmol) at RT. The reaction mixture was stirred in a 30 ml sealed tube at 45 °C for 16 hours The reaction mixture was concentrat ed to give a residue. The residue was purified by preparative HPLC [Instrument:ACSWH‐GX‐C; ColumnPhenomenex luna C18 150*25mm* 10µm; eluent A: water (0.225% formic acid in water), eluent B: acetonitrile; gradient: 0‐10 m in 25‐55% B; flow 25 ml/min; temperature: RT; Detector: UV 220/254 nm.] to give the title compound (55 mg, 58%). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.617 (6 .33), 1.634 (6.55), 2.324 (0.62), 2.452 (16.00), 2.66 6 (0.48), 5.629 (1.09), 5.647 (1.63), 5.665 (1.14), 7.5 45 (0.77), 7.563 (2.11), 7.587 (3.72), 7.731 (2.32), 7.764 (2.24), 7.782 (1.94), 7.852 (3.38), 8.211 (2.19), 8.379 (0.55), 8.952 (4.68), 9.055 (4.64), 9.213 (1.76), 9.231 (1.82). Example 321 N,2‐dimethyl‐4‐({(1R)‐1‐[3‐(trifluoromethyl)phen yl]ethyl}amino)pyrido[3,4‐d]pyrimidine‐6‐ carboxamide
To a solution of methylamine in ethanol (2 M) was added Example 319 (120 mg, 307 µmol) at room temperature. The reaction mixture was heated in a se aled tube at 40 °C for 16 hours. The reaction mixture was concentrated to give a residue. The residue was purified by preparative HPLC [Instrument:ACSWH‐GX‐C; Column: Phenomenex Luna C18 150*25mm*10µm; eluent A: water (0.225% formic acid in water), eluent B: acetonitrile; gradient: 0‐10 m in 25‐55% B; flow 25 ml/min; temperature: RT; Detector: UV 220/254 nm.] to give t he title compound (32 mg, 26%). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.624 (6 .20), 1.642 (6.35), 2.452 (16.00), 2.864 (7.76), 2.87 6 (8.04), 5.633 (0.99), 5.651 (1.50), 5.669 (1.01), 7.5 47 (0.60), 7.566 (1.94), 7.585 (3.48), 7.589 (3.39), 7.609 (0.81), 7.768 (1.81), 7.785 (1.49), 7.856 (3.12), 8.407 (0.54), 8.858 (1.47), 8.870 (1.49), 8.956 (5.09), 9.023 (4.83), 9.241 (1.66), 9.260 (1.65). Example 322 1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐methylphen yl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]piperidine‐4‐carbonitrile Using the method described for Example 25: Intermediate 16 (50 mg, 186 µmol) was treated with (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐ 1‐amine hydrochloride (49.4 mg, 223 µmol) and gave the titled compound (53 mg, 62%) after preparat ive HPLC purification (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.544 (4.76), 1.561 (4.83), 1.802 (0.71), 1.808 (0.73), 1.831 (0.85), 1.854 (0.41), 2.004 (0.84), 2.318 (16.00), 2. 518 (1.48), 2.523 (1.21), 2.534 (7.42), 3.121 (0.45), 3.131 (0.63), 3.142 (0.85), 3.153 (0.62), 3.164 (0.41), 3.385 (0.68), 3.411 (1.02), 3.438 (0.72), 3.873 (0.86), 3.906 (0.76), 5.715 (0.69), 5.732 (1.05), 5.7 50 (0.68), 7.078 (0.91), 7.216 (1.93), 7.282 (0.65), 7.300 (1.63), 7.320 (1.13), 7.353 (0.79), 7.382 (1.64), 7.400 (1.05), 7.462 (2.93), 7.623 (1.26), 7.641 (1.12), 8.464 (1.13), 8.483 (1.09), 8.645 (4.51). Example 323
N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl] ethyl}‐6‐[(2S)‐2,4‐dimethylpiperazin‐1‐yl]‐2 methylpyrido[3,4‐d]pyrimidin‐4‐amine Using the method described for Example 25: Intermedia te 17 (35.0 mg, 128 µmol) was treated with (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐ 1‐amine hydrochloride (34.7 mg, 154 µmol) and gave the titled compound (51 mg, 86%) after preparat ive HPLC purification (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.859 (0.88), 0.967 (2.64), 1.109 (2.22), 1.132 (5.97), 1.144 (2.86), 1.149 (6.06), 1.208 (0.45), 1.224 (0.58), 1.6 03 (5.21), 1.621 (5.18), 1.989 (0.45), 2.010 (0.70), 2.017 (0.72), 2.038 (0.47), 2.164 (0.75), 2.173 (0.87 ), 2.192 (0.92), 2.201 (0.87), 2.230 (10.75), 2.298 (16.00), 2.318 (0.49), 2.323 (0.92), 2.327 (1.22), 2. 331 (0.87), 2.336 (0.40), 2.518 (5.31), 2.523 (3.58), 2.660 (0.41), 2.665 (0.87), 2.669 (1.20), 2.673 (0.83), 2.747 (1.05), 2.774 (0.96), 2.903 (0.73), 2.931 (0.68), 3.070 (0.49), 3.078 (0.70), 3.101 (0.85), 3.1 09 (0.77), 3.132 (0.49), 3.164 (0.51), 3.906 (0.70), 3.937 (0.64), 4.684 (0.62), 5.757 (0.77), 5.775 (1.19), 5.793 (0.77), 7.103 (1.19), 7.240 (2.48), 7.278 (0.87), 7.298 (1.92), 7.317 (1.13), 7.344 (3.07), 7.3 75 (1.05), 7.487 (0.66), 7.504 (1.13), 7.522 (0.55), 7.635 (0.62), 7.653 (1.11), 7.672 (0.55), 8.411 (1.24 ), 8.430 (1.20), 8.663 (4.82). Example 324 {1‐[4‐({(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophe nyl]ethyl}amino)‐2‐methylpyrido[3,4‐d]pyrimidin‐6‐ yl]‐ 4‐methylpiperazin‐2‐yl}methanol (mixture of stereoi somers) Using the method described for Example 25: Intermedia te 18 (33.0 mg, 145 µmol) was treated with (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐ 1‐amine hydrochloride (34.7 mg, 154 µmol) and gave the titled compound (32 mg, 55%) after preparat ive HPLC purification (basic method). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.967 (0.55), 1.109 (1.44), 1.598 (4.22), 1.605 (4.42), 1.616 (4.36), 1.623 (4.10), 1.974 (0.46), 2.004 (1.93), 2.0 14 (1.28), 2.023 (1.17), 2.032 (1.55), 2.233 (14.08), 2.290 (11.34), 2.296 (11.41), 2.518 (16.00), 2.523 (10.44), 2.673 (2.26), 2.877 (0.98), 2.903 (0.91), 3.072 (0.53), 3.109 (2.04), 3.136 (1.60), 3.779 (0.79), 3.919 (0.51), 3.950 (0.54), 3.998 (0.45), 4.550
(0.76), 4.741 (0.81), 4.753 (0.83), 5.747 (0.66), 5.755 (0.74), 5.765 (1.03), 5.773 (1.04), 5.783 (0.71 ), 5.791 (0.65), 7.103 (1.22), 7.239 (2.48), 7.272 (0.69), 7.281 (0.72), 7.291 (1.48), 7.300 (1.51), 7.310 (0.92), 7.319 (0.87), 7.341 (2.18), 7.361 (2.16), 7.3 74 (1.22), 7.502 (1.31), 7.635 (0.80), 7.653 (1.42), 7.671 (0.72), 8.408 (0.96), 8.420 (1.19), 8.437 (0.92 ), 8.633 (3.50), 8.640 (3.50). Example 325 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐ dihydroimidazo[1,2‐a]pyrazin‐7(8H)‐yl]pyrido[3,4‐d]p yrimidin‐4‐amine Using the method described for Example 25: Intermediate 19 (30 mg, 86 µmol) was treated with (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐ 1‐amine hydrochloride (23 mg, 103 µmol) and gave the titled compound (15 mg, 33%) after preparative H PLC purification (basic method). 1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (3.17), 1.225 (0.55), 1.348 (0.42), 1.632 (5.40), 1.650 (5.34), 2.325 (16.00), 2.518 (6.33), 2.523 (4.14), 2. 660 (0.42), 2.665 (0.91), 2.669 (1.29), 2.673 (0.93), 2.678 (0.42), 4.205 (2.79), 4.215 (3.02), 4.224 (2.01), 4.825 (3.65), 4.830 (3.69), 5.760 (0.82), 5.778 (1.25), 5.796 (0.80), 7.107 (1.20), 7.243 (2.51), 7.2 85 (0.91), 7.303 (2.01), 7.323 (1.14), 7.379 (1.06), 7.495 (0.70), 7.512 (1.18), 7.530 (0.57), 7.623 (3.25), 7.648 (0.68), 7.666 (1.18), 7.684 (0.59), 7.836 (3.00), 7.839 (3.15), 8.490 (1.33), 8.509 (1.29), 8.7 41 (4.92). Example 326 N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐methyl‐6‐[2‐(trifluoromethyl)‐5,6‐ dihydro[1,2,4]triazolo[1,5‐a]pyrazin‐7(8H)‐yl]pyrido[3 ,4‐d]pyrimidin‐4‐amine Using the method described for Example 25: Intermediate 20 (30 mg, 85 µmol) was treated with (1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethan‐ 1‐amine hydrochloride (23 mg, 102 µmol) and gave the titled compound (15 mg, 31%) after preparative H PLC purification (basic method).
1 H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.636 (5 .12), 1.653 (5.03), 2.330 (16.00), 2.518 (7.79), 2.52 3 (4.89), 2.665 (1.21), 2.669 (1.66), 2.673 (1.18), 4.3 02 (1.12), 4.314 (2.28), 4.328 (1.69), 4.441 (1.49), 4.455 (2.14), 4.983 (4.47), 5.762 (0.73), 5.779 (1.18), 5.797 (0.76), 7.108 (1.12), 7.243 (2.39), 7.286 (0.87), 7.305 (1.91), 7.324 (1.10), 7.379 (1.01), 7.4 98 (0.67), 7.515 (1.12), 7.532 (0.56), 7.649 (0.65), 7.669 (1.29), 7.679 (3.32), 8.488 (1.27), 8.506 (1.21 ), 8.757 (4.72). Example 327 6‐(cyclobutyloxy)‐N‐{(1R)‐1‐[3‐(difluoromethyl) 2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine To sodium hydride (60% dispersion on mineral oil, 28.5 mg, 714 µmol) under Argon was added a solution of cyclobutanol (51.5 mg, 714 µmol) in NMP (2 ml) and stirred for 5 min at RT. Then Example 2 (50 mg, 143 µmol) was added and the reaction he ated using a microwave at 180°C for 20 min. The reaction mixture was diluted with water and extracted with EtOAc. The combined organics were washed with sat. NaCl, filtered through a hydrophobic filter and concentrated. The title compound (6.7 mg, 12%) was isolated after preparative HPLC purification along with the ring‐opened side‐ product (see Example 328, 1.5 mg, 3%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (0.46), 1.228 (0.71), 1.394 (0.41), 1.418 (0.75), 1.443 (0.87), 1.463 (0.58), 1.495 (5.88), 1.512 (5.90), 1.6 95 (0.54), 1.705 (0.75), 1.721 (0.61), 2.298 (0.59), 2.318 (1.78), 2.323 (1.71), 2.327 (1.61), 2.340 (2.65), 2.361 (1.63), 2.364 (1.64), 2.386 (0.99), 2.412 (16.00), 2.518 (3.94), 2.523 (2.71), 2.539 (0.75), 2. 665 (0.48), 2.669 (0.66), 2.673 (0.48), 4.715 (0.89), 4.736 (1.24), 4.754 (0.86), 5.729 (0.83), 5.746 (1.29), 5.764 (0.84), 6.962 (1.03), 7.100 (2.00), 7.205 (1.11), 7.224 (2.48), 7.238 (1.12), 7.243 (1.55), 7.4 37 (1.61), 7.456 (1.34), 7.617 (1.47), 7.635 (1.35), 8.218 (2.88), 8.744 (4.28), 8.767 (1.39), 8.785 (1.35 ). Example 328 6‐butoxy‐N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐flu orophenyl]ethyl}‐2‐methylpyrido[3,4‐d]pyrimidin‐4‐ amine Isolated as a side‐product (see Example 327). 1H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 0.832 (0.50), 0.850 (0.77), 0.947 (4.12), 0.965 (9.91), 0.984 (4.82), 1.229 (2.06), 1.347 (0.53), 1.496 (1.46), 1.5 11 (6.55), 1.528 (6.09), 1.553 (1.00), 1.807 (0.57), 1.824 (1.56), 1.842 (2.13), 1.859 (1.36), 1.878 (0.50), 2.322 (1.03), 2.326 (1.20), 2.331 (0.86), 2.382 (16.00), 2.412 (2.06), 2.522 (4.32), 2.664 (0.77), 2. 669 (1.03), 2.673 (0.77), 2.692 (0.60), 2.722 (0.53), 2.856 (0.43), 3.300 (0.47), 3.898 (0.47), 3.913 (1.06), 3.936 (1.13), 3.952 (0.47), 4.425 (0.50), 4.441 (1.16), 4.464 (1.10), 4.481 (0.47), 5.814 (0.83), 5.8 31 (1.26), 5.850 (0.83), 6.999 (1.03), 7.136 (1.96), 7.236 (1.23), 7.256 (2.40), 7.275 (2.16), 7.458 (1.73), 7.477 (1.33), 7.664 (1.46), 7.683 (1.36), 8.201 (2.86), 8.218 (0.43), 8.738 (4.39), 8.776 (1.33), 8.7 94 (1.26). Table 15: Examples 329‐337 Following the method described here for Example 327, the following Examples in Table 15 were prepared either as their mono‐ or di‐substituted analogs.
Example 338 6‐[(azetidin‐3‐yl)oxy]‐N‐{(1R)‐1‐[3‐(difluor omethyl)‐2‐fluorophenyl]ethyl}‐2‐methylpyrido[3,4‐ d]pyrimidin‐4‐amine hydrochloride x.HCl To a solution of Example 332 (13.4 mg, 26.6 µmol) in dioxane (130 µl) was added a HCl solution in dioxane (4M, 130 µmol) and stirred at RT for 1h. The reaction was concentrated to give the title compound (13 mg). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (16.00 ), 1.232 (0.51), 1.593 (0.96), 1.669 (2.47), 1.686 (2.49), 1.709 (1.27), 1.727 (1.17), 1.907 (0.50), 2.3 32 (0.76), 2.423 (6.04), 2.431 (6.05), 2.518 (4.08), 2.523 (2.61), 2.579 (2.11), 2.673 (0.75), 3.384 (0.82), 3.675 (0.45), 4.064 (0.40), 5.248 (0.50), 5.281 (0.40), 5.706 (0.40), 5.792 (0.53), 5.810 (0.53), 7.0 96 (0.51), 7.103 (0.78), 7.231 (1.04), 7.238 (1.62), 7.290 (0.44), 7.307 (0.96), 7.326 (0.57), 7.357 (0.73), 7.367 (0.58), 7.375 (1.10), 7.512 (0.53), 7.529 (0.88), 7.549 (0.49), 7.573 (0.43), 7.826 (0.44), 8.6 20 (0.84), 8.879 (0.51), 9.557 (3.01). Example 339 tert‐butyl {(3‐trans)‐1‐[4‐({(1R)‐1‐[3‐(dif luoromethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐ methylpyrido[3,4‐d]pyrimidin‐6‐yl]‐4‐fluoropyrroli din‐3‐yl}carbamate (mixture of stereoisomers) To a solution of Example 2 (50.0 mg, 143 µmol) in DMSO (1.3 ml) was added tert‐butyl [rac‐(trans) 4‐ fluoropyrrolidin‐3‐yl]carbamate (58.3 mg, 285 µmol) and TEA (80 µl, 570 µmol). The reaction was heated at 110°C for 16h. Another portion of the am ine was added (58.3 mg, 285 µmol) and TEA (80 µl, 570 µmol) were added and heated at 130°C for 16h. The reaction was allowed to cool and then purified by preparative HPLC (basic method) to give the titled compound (23 mg, 28%). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (2.26) , 1.404 (16.00), 1.612 (4.71), 1.630 (4.52), 2.273 (0.50), 2.300 (10.47), 2.327 (0.45), 2.401 (0.69), 2. 518 (1.82), 2.522 (1.09), 2.725 (0.51), 3.489 (0.51), 3.501 (0.49), 3.517 (0.61), 3.746 (0.64), 3.762 (0.74), 3.776 (1.11), 3.803 (0.71), 5.155 (0.55), 5.284 (0.55), 5.763 (0.55), 5.780 (0.82), 5.796 (0.53), 7.1 02 (1.09), 7.148 (2.78), 7.237 (2.25), 7.273 (0.75), 7.293 (1.64), 7.312 (0.97), 7.373 (0.96), 7.454 (0.61), 7.469 (0.61), 7.486 (0.71), 7.503 (1.08), 7.521 (0.54), 7.632 (0.54), 7.650 (0.99), 7.668 (0.55), 8.4 09 (1.13), 8.427 (1.10), 8.655 (3.98). Example 340 6‐[(trans)‐3‐amino‐4‐fluoropyrrolidin‐1‐yl]‐ N‐{(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]eth yl}‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride ( mixture of stereoisomers) Using the method described for Example 338: Example 339 (17.1 mg, 32.0 µmol) gave the titled compound (16.6 mg). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (16.00 ), 1.143 (0.40), 1.223 (1.20), 1.231 (0.61), 1.740 (3.25), 1.757 (3.26), 2.323 (0.47), 2.327 (0.65), 2.3 32 (0.46), 2.518 (2.97), 2.523 (2.22), 2.537 (7.42), 2.665 (0.46), 2.669 (0.64), 2.673 (0.45), 2.737 (0.40), 3.841 (0.68), 3.919 (0.65), 3.934 (1.02), 3.950 (0.76), 3.965 (0.69), 3.982 (0.41), 4.160 (0.55), 5.5 10 (0.59), 5.634 (0.61), 5.983 (0.68), 5.992 (0.63), 6.000 (0.48), 7.109 (0.92), 7.244 (1.87), 7.338 (0.87), 7.357 (1.88), 7.378 (1.79), 7.551 (0.73), 7.568 (1.21), 7.585 (0.59), 7.941 (0.73), 8.730 (0.89), 8.7 90 (0.74), 8.844 (1.61), 8.860 (1.31). Example 341 tert‐butyl {(cis)‐1‐[4‐({(1R)‐1‐[3‐(difluorom ethyl)‐2‐fluorophenyl]ethyl}amino)‐2‐methylpyrido[3, 4‐ d]pyrimidin‐6‐yl]‐4‐fluoropyrrolidin‐3‐yl}carbam ate (mixture of stereoisomers) Using the method described for Example 339: Example 2 (17.1 mg, 32.0 µmol) treated with tert‐butyl [rac‐(cis)‐4‐fluoropyrrolidin‐3‐yl]carbamate (58.3 mg, 285 µmol) gave the titled compound (16 mg, 20%) after preparative HPLC purification (basic method ). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.231 (0.46) , 1.433 (16.00), 1.608 (2.37), 1.626 (2.37), 2.294 (4.13), 2.298 (4.49), 2.326 (0.43), 2.518 (1.77), 2.5 22 (1.09), 2.669 (0.42), 3.314 (0.59), 3.828 (0.65), 3.847 (0.68), 3.870 (0.50), 5.774 (0.60), 7.102 (0.61), 7.121 (1.22), 7.238 (1.26), 7.291 (0.68), 7.374 (0.67), 7.400 (0.42), 7.502 (0.51), 7.647 (0.56), 8.4 02 (0.48), 8.420 (0.48), 8.648 (2.34). Example 342 6‐[(cis)‐3‐amino‐4‐fluoropyrrolidin‐1‐yl]‐N {(1R)‐1‐[3‐(difluoromethyl)‐2‐fluorophenyl]ethyl }‐2‐ methylpyrido[3,4‐d]pyrimidin‐4‐amine hydrochloride ( mixture of stereoisomers) Using the method described for Example 338: Example 341 (13.3 mg, 24.9 µmol) gave the titled compound (13 mg). ¹H‐NMR (400 MHz, DMSO‐d6) δ [ppm]: 1.107 (16.00 ), 1.224 (1.24), 1.232 (0.79), 1.731 (3.65), 1.748 (3.62), 2.323 (0.77), 2.327 (1.07), 2.332 (0.77), 2.5 18 (7.03), 2.523 (8.51), 2.665 (0.79), 2.669 (1.11), 2.673 (0.77), 3.504 (0.41), 3.526 (0.77), 3.542 (0.81), 3.565 (0.45), 3.899 (0.45), 3.935 (0.69), 3.965 (0.52), 3.974 (0.49), 3.999 (0.41), 4.064 (0.52), 4.0 88 (0.69), 4.109 (0.58), 5.482 (0.64), 5.620 (0.62), 5.953 (0.47), 5.969 (0.67), 5.985 (0.47), 7.107 (1.01), 7.243 (2.10), 7.336 (0.69), 7.355 (1.52), 7.378 (1.42), 7.552 (0.69), 7.569 (1.18), 7.587 (0.60), 7.8 91 (0.58), 8.827 (2.34).
EXPERIMENTAL SECTION – BIOLOGICAL ASSAYS Examples were tested in selected biological assays on e or more times. When tested more than once, data are reported as either average values or as me dian values, wherein ^ the average value, also referred to as the arithmeti c mean value, represents the sum of the values obtained divided by the number of times teste d, and ^ the median value represents the middle number of the group of values when ranked in ascending or descending order. If the number of valu es in the data set is odd, the median is the middle value. If the number of values in the d ata set is even, the median is the arithmetic mean of the two middle values. Examples were synthesized one or more times. When synthesized more than once, data from biological assays represent average values or median values calculated utilizing data sets obtained from testing of one or more synthetic batch. In vitro metabolic stability in human liver microsomes. The in vitro metabolic stability of test compounds was determined by incubating them at 1 µM in a suspension of liver microsomes in 100 mM phosphate buffer, pH 7.4 (NaH2PO4 x H2O + Na2HPO 4 x 2H2O) and at a protein concentration of 0.5 mg/mL at 37 °C. The microsomes were activat ed by adding a co‐factor mix containing 8 mM Glucose‐6‐phosphate, 4 mM MgCl2, 0.5 mM NADP and 1 IU/ml G‐6‐P‐Dehydrogenase in phosphate buffer, pH 7.4. The metabolic assay was started shor tly afterwards by adding the test compound to the incubation at a final volume of 1 mL. Organic solve nt in the incubations was limited to ≤0.01 % dimethylsulfoxide (DMSO) and ≤1% acetonitrile. During incubation, the microsomal suspensions were continuously shaken at 580 rpm and aliquots were taken at 2, 8, 16, 30, 45 and 60 min, to which equal volumes of cold methanol were immediately added. Samples were frozen at ‐20 °C overnight, subsequently centrifuged for 15 minutes at 3000 rpm and the supernatant was analyzed with an Agilent 1200 HPLC‐system with LC/MS‐MS de tection. The half‐life of a test compound was determined from the concentration‐time plot. From the half‐life the intrinsic clearances and the hepatic in vivo blood clearance (CL) and maximal ora l bioavailability (Fmax) were calculated using the ‘well stirred’ liver model together with the additional parameters liver blood flow, specific liver weight and microsomal protein content. The following parameter values were used: Liver blood flow 1.32 L/h/kg, specific liver weight 21 g/kg, microsoma l protein content 40 mg/g. In vitro metabolic stability in rat hepatocytes. Hepatocytes from Han/Wistar rats were isolated via a 2‐step perfusion method. After perfusion, the liver was carefully removed from the rat: the liver capsule was opened and the hepatocytes were gently shaken out into a Petri dish with ice‐cold Williams’ medium E (WME). The resulting cell
suspension was filtered through sterile gaze in 50 ml falcon tubes and centrifuged at 50 × g for 3 min at room temperature. The cell pellet was resuspended in 30 ml WME and centrifuged twice through a Percoll® gradient at 100 × g. The hepatocytes w ere washed again with WME and resuspended in medium containing 5 % FCS. Cell viability was determ ined by trypan blue exclusion. For the metabolic stability assay liver cells were distributed in WME containing 5 % FCS to glass vials at a density of 1.0 × 106 vital cells/ml. The test compound was added to a final concentration of 1 µM. During incubation, the hepatocyte suspensions were continuously shaken at 580 rpm and aliquots were taken at 2, 8, 16, 30, 45 and 90 min, to which equal volumes of cold methanol were immediately added. Samples were frozen at ‐20 °C overnight, subsequently centrifuged for 15 minutes at 3000 rpm and the supernatant was analyzed with an Agilent 1200 HPLC‐system with LC/MS‐MS detection. The half‐life of a test compound was determined fr om the concentration‐time plot. From the half‐life the intrinsic clearances and the hepatic in vivo blo od clearance (CL) and maximal oral bioavailability (Fmax) were calculated using the ‘well stirred’ liver model together with the additional parameters liver blood flow, specific liver weight and amount of liver cells in vivo and in vitro. The following parameter values were used: Liver blood flow 4.2 L/h /kg, specific liver weight 32 g/kg, liver cells in vivo 1.1 x 108 cells/g liver, liver cells in vitro 1.0 x 106/ml. Caco‐2 Permeability Assay. Caco‐2 cells (purchased from DSMZ Braunschweig, Germ any) were seeded at a density of 4.5 × 10 4 cells/well on 24‐well insert plates, 0.4 µm pore size, and grown for 15 d in DMEM supplemented with 10% FCS, 1% GlutaMAX (100 ×, Gibco), 100 U/mL peni cillin, 100 µg/mL streptomycin (Gibco) and 1% non‐essential amino acids (100 ×). Cells were maintained at 37 °C in a humidified 5% CO 2 atmosphere. Medium was changed every 2–3 d. Before the permeation assay was run, the culture medium was replaced by FCS‐free HEPES carbonate tra nsport buffer (pH 7.2) For the assessment of monolayer integrity, the transepithelial electrical resistance was measured. Test compounds were predissolved in DMSO and added either to the apical or basolateral compartment at a final concentration of 2 µM. Before and after incubation for 2 h at 37 °C, samples were taken f rom both compartments and analyzed by LC‐MS/MS after precipitation with MeOH. Permeability (P app ) was calculated in the apical to basolateral (A → B) and basolateral to apical (B → A) direcƟons. The apparent permeability was calculated using following e quation: P app = (V r /P 0 )(1/S)(P 2 /t), where V r is the volume of medium in the receiver chamber, P 0 is the measured peak area of the test drug in the donor chamber at t = 0, S is the surface area of the monolayer, P 2 is the measured peak area of the test drug in the acceptor chamber after incubation for 2 h, and t is the incubation time. The efflux ratio basolateral (B) to apical (A) was calculated as P app B–A/P app A–B. In addition, the compound recovery was calculated. As an assay control, referen ce compounds were analyzed in parallel.
6,7‐dimethoxy‐N‐[(1R)‐1‐(1‐naphthyl)ethyl]qui nazolin‐4‐amine, which was used to calibrate the assay, was prepared as follows: To 4‐chloro‐6,7‐dimethoxyquinazoline (100 mg, 0.44 5 mmol, commercially available) in 1.7 mL DMSO was added (1R)‐1‐(1‐naphthyl)ethanamine (76 mg, 0 .445 mmol, commercially available) and N‐ethyl‐ N‐isopropylpropan‐2‐amine (202 µl, 1.16 mmol). The reaction was stirred at 100°C overnight, cooled to ambient temperature and filtered. After removal of the solvent under reduced pressure the crude product was purified via HPLC chromatography to yield the title compound (118 mg, 73%). 1 H‐NMR (400 MHz ,DMSO‐d6), d [ppm]= 1.72 (3H), 3.90 (6H), 6.32‐6.41 (1H), 7.09 (1H), 7.46‐7.58 (3H), 7.64 7.69 (1H), 7.78 (2H), 7.92‐7.97 (1H), 8.18‐8.24 ( 2H), 8.28 (1H). The in vitro activity of the compounds of the prese nt invention can be demonstrated in the following assays: Biochemical assay 1: hK‐RasG12C interaction assay wi th hSOS1 This assay quantifies the equilibrium interaction of human SOS1 (hSOS1) with human K‐Ras G12C (hK‐ RasG12C). Detection of the interaction is achieved by measuring homogenous time‐resolved fluorescence resonance energy transfer (HTRF) from ant iGST‐Europium (FRET donor) bound to GST‐ K‐RasG12C to anti‐6His‐XL665 bound to His‐tagge d hSOS1 (FRET‐acceptor). The assay buffer containes 5 mM HEPES pH 7.4 (Applichem), 150 mM NaCl (Sigma), 10 mM EDTA (Promega), 1 mM DTT (Thermofisher), 0.05% BSA Fractio n V, pH 7.0, (ICN Biomedicals), 0.0025% (v/v) Igepal (Sigma) and 100 mM KF (FLUKA). The expression and purification of N‐terminal GST‐tagged hK‐RasG12C and N‐terminal His‐tagged hSOS1 is described below. Concentrations of protein batches used are optimized to be within the linear range of the HTRF signal. A Ras working solu tion is prepared in assay buffer containing typically 10 nM GST‐hK‐RasG12C and 2 nM antiGST‐Eu(K) (Cisbio, France). A SOS1 working solution is prepared in assay buffer containing typically 20nM His‐hSOS1 and 10 nM anti‐6His‐XL665 (Cisbio, France). An inhibitor control solution is prepared in assay buffer containing 10 nM anti‐6His‐XL665 without hSOS1.
Fifty nl of a 100‐fold concentrated solution of the test compound in DMSO are transferred into a black microtiter test plate (384 or 1536, Greiner Bi o‐One, Germany). For this, either a Hummingbird liquid handler (Digilab, MA, USA) or an Echo acousti c system (Labcyte, CA, USA) is used. All steps of the assay are performed at 20°C. A v olume of 2.5 µl of the Ras working solution is ad ded to all wells of the test plate using a Multidrop dispenser (Thermo Labsystems). After 2 min preincubation, 2.5 µl of the SOS1 working solution are added to all wells except for those wells at t he side of the test plate that are subsequently filled with 2.5 µl of the inhibitor control solution. Aft er 60 min incubation the fluorescence is measured with a Pherastar (BMG, Germany) using the HTRF module (excitation 337nm, emission 1: 620nm, emission 2: 665nm). The ratiometric data (emission 2 divided by emission 1) are normalized using the controls (DMSO = 0% inhibition, inhibition control wells with inhibitor control solution = 100% inhibition). Compounds are tested in duplicates at up to 11 concentrations (for example 20 µM, 5,7 µM, 1,6 µM, 0,47 µM, 0,13 µM, 38 nM, 11 nM, 3,1 nM, 0,89 nM, 0,25 nM and 0,073 nM). IC50 values are calculated by 4‐ Parameter fitting using a commercial software package (Genedata Screener, Switzerland). Biochemical assay 2: hK‐RasG12C activation assay by hSOS1 at high GTP concentration This assay quantifies human SOS1‐mediated nucleotide exchange of human K‐Ras G12C (hK‐RasG12C) preloaded with a fluorescent GTP‐analog and in presence of an excess of free GTP. Loaded hK‐ RasG12C generates a high HTRF‐signal by energy tran sfer from antiGST‐Terbium (FRET donor) bound to hK‐Ras to the loaded fluorescent GDP analog (FRET‐acceptor). hSOS1 activity exchanges the fluorescent GDP for non‐fluorescent GTP and therefor e leads to a reduction of the HTRF signal. The fluorescent GDP‐analog EDA‐GDP‐Dy647P1 (2’/3’‐O‐(2‐Aminoethyl‐carbamoyl)‐guanosine‐ 5’‐ diphosphate labelled with Dy647P1 (Dyomics GmbH, Germany)) is synthesized by Jena Biosciences GmbH (Germany) and supplied as a 1mM aqueous solutio n. The expression and purification of N‐terminal GST‐tagged human K‐RasG12C and N‐terminal His‐ tagged human SOS1 is described below. Concentrations of protein batches used are optimized to be within the linear range of the HTRF signal. Preparation of GST‐tagged hK‐RasG12C loaded with fluorescent nucleotide is performed as follows: incubation of 11.5 µM hK‐Ras G12C with 5‐fold excess GDP‐Dy647 nucleotide (54 µM) in 500 µl NLS‐ buffer (RAS activation Kit Jena Bioscience, Kat. #PR‐950) for 10 min at 37°C. Addition of 20 µl 1 M MgCl 2 (Sigma) to final 40 mM and store on ice. Purification into buffer (10 mM HEPES pH 7.4 (Applichem), 150 mM NaCl (Sigma), 5 mM MgCl 2 (Sigma)) by use of a PD‐Minitrap desalting column (GE Healthcare). Concentration of 1 ml purified hK‐ Ras‐GDP‐Dy647 is approx. 4‐5 µM.
The assay buffer containes 10 mM HEPES pH 7.4 (Applichem), 150 mM NaCl (Sigma), 5 mM MgCl 2 (Sigma), 1 mM DTT (Thermofisher), 0.05% BSA Fraction V, pH 7.0, (ICN Biomedicals), 0.0025% (v/v) Igepal (Sigma). A Ras working solution is prepared in assay buffer containing typically 80 nM loaded GST‐hK‐ RasG12C‐EDA‐GDP‐Dy647P1 and 2 nM antiGST‐Tb (Cisbio, France). A hSOS1 working solution is prepared in assay buffer containing typically 8nM His‐hSOS1 and 100 µM GTP (Jena Bioscience, Germany). An inhibitor control solution is prepared in assay buffer containing the same concentration of hSOS1 without GTP. Alternatively, the inhibitor control solution is prepa red by supplementing the hSOS1 working solution with 20 µM of 6,7‐dimethoxy‐N‐[(1R)‐1‐(1‐naphthyl)ethyl]quinaz olin‐4‐amine which is used to calibrate the assay. Fifty nl of a 100‐fold concentrated solution of the test compound in DMSO are transferred into a black microtiter test plate (384 or 1536, Greiner Bi o‐One, Germany). For this, either a Hummingbird liquid handler (Digilab, MA, USA) or an Echo acousti c system (Labcyte, CA, USA) is used. All steps of the assay are performed at 20°C. A v olume of 2.5 µl of the Ras working solution is ad ded to all wells of the test plate using a Multidrop dispenser (Thermo Labsystems). After 2 min preincubation, 2.5 µl of the hSOS1 working solution are added to all wells except for those wells at the side of the test plate that are subsequently filled with 2.5 µl of the inhibitor control solution. After 20 min incubation the fluorescence is measured with a Pherastar (BMG, Germany) using the HTRF module (excitation 337nm, emission 1: 620nm, emi ssion 2: 665nm). The ratiometric data (emission 2 divided by emission 1) are normalized using the controls (DMSO = 0% inhibition, inhibition control wells with inhibitor control solution = 100% inhibition). Compounds are tested in duplicates at up to 11 concentrations (for example 20 µM, 5,7 µM, 1,6 µM, 0,47 µM, 0,13 µM, 38 nM, 11 nM, 3,1 nM, 0,89 nM, 0,25 nM and 0,073 nM). IC50 values are calculated by 4‐ Parameter fitting using a commercial software package (Genedata Screener, Switzerland). Biochemical assay 3: hK‐RasG12C activation assay by hSOS1 K‐Ras is a small GTPase that can bind GDP and GTP. The guanine nucleotide e xchange factor SOS1 catalyzes the activation of K‐Ras by promoting the exchange of GDP to GTP. SOS1 binds to K‐Ras‐GDP thereby opening the GDP‐binding pocket to facilitate GDP release. Rebinding of excess nucleotide leads to dissociation of the K‐Ras‐SOS1 intermediate complex leaving K‐Ras loaded with the nucleotide. This assay quantifies human SOS1‐ (hSOS1‐) mediate d loading of human K‐Ras G12C ‐GDP (hK‐RasG12C‐ GDP) with a fluorescent GTP‐analog. Detection of successful loading is achieved by measuring
homogenous time‐resolved fluorescence resonance energy transfer (HTRF) from antiGST‐Terbium (FRET donor) bound to GST‐hK‐RasG12C (see below) to the loaded fluorescent GTP analog (FRET‐ acceptor). The fluorescent GTP‐analog EDA‐GTP‐Dy647P1 (2’/3’‐O‐(2‐Aminoethyl‐carbamoyl)‐guanosine‐ 5’‐ triphosphate labelled with Dy647P1 (Dyomics GmbH, Germany)) is synthesized by Jena Biosciences GmbH (Germany) and supplied as a 1mM aqueous solutio n. The assay buffer containes 10 mM HEPES pH 7.4 (Applichem), 150 mM NaCl (Sigma), 5 mM MgCl 2 (Sigma), 1 mM DTT (Thermofisher), 0.05% BSA Fraction V, pH 7.0, (ICN Biomedicals), 0.0025% (v/v) Igepal (Sigma). The expression and purification of N‐terminal GST‐tagged human K‐RasG12C and N‐terminal His‐ tagged hSOS1 is described below. Concentrations of pr otein batches used are optimized to be within the linear range of the HTRF signal. A hRas working solution is prepared in assay buffer containing typically 100 nM GST‐hK‐RasG12C and 2 nM antiGST Tb (Cisbio, France). A hSOS1 working solution is prepared in assay buffer containing typically 20nM hSOS1 and 200 nM EDA‐GTP‐Dy647P1. An inhibitor control solution is prepared in assay buffer containing 200 nM EDA‐GTP‐Dy647P1 without hSOS1. Fifty nl of a 100‐fold concentrated solution of the test compound in DMSO are transferred into a black microtiter test plate (384 or 1536, Greiner Bi o‐One, Germany). For this, either a Hummingbird liquid handler (Digilab, MA, USA) or an Echo acousti c system (Labcyte, CA, USA) is used. All steps of the assay are performed at 20°C. A v olume of 2.5 µl of the hRas working solution is a dded to all wells of the test plate using a Multidrop dispenser (Thermo Labsystems). After 10 min preincubation, 2.5 µl of the hSOS1 working solution are added to all wells except for those wells at the side of the test plate that are subsequently filled with 2.5 µl of the inhibitor control solution. After 30 min incubation the fluorescence is measured with a Pherastar (BMG, Germany) using the HTRF module (excitation 337nm, emission 1: 620nm, emi ssion 2: 665nm). The ratiometric data (emission 2 divided by emission 1) are normalized using the controls (DMSO = 0% inhibition, inhibition control wells with inhibitor control solution = 100% inhibition). Compounds are tested in duplicates at up to 11 concentrations (for example 20 µM, 5,7 µM, 1,6 µM, 0,47 µM, 0,13 µM, 38 nM, 11 nM, 3,1 nM, 0,89 nM, 0,25 nM and 0,073 nM). IC50 values are calculated by 4‐ Parameter fitting using a commercial software package (Genedata Screener, Switzerland). Biochemical assay 4: hK‐RasG12C activation assay by hSOS2 This assay quantifies hSOS2‐mediated loading of hK‐Ras G12C ‐GDP (hK‐RasG12C‐GDP) with a fluorescent GTP‐analog. Detection of successful loadi ng is achieved by measuring homogenous time‐
resolved fluorescence resonance energy transfer (HTR F) from antiGST‐Terbium (FRET donor) bound to GST‐hK‐RasG12C to the loaded fluorescent GTP a nalog (FRET‐acceptor). The fluorescent GTP‐analog EDA‐GTP‐Dy647P1 (2’/3’‐O‐(2‐Aminoethyl‐carbamoyl)‐guanosine‐ 5’‐ triphosphate labelled with Dy647P1 (Dyomics GmbH, Germany)) is synthesized by Jena Biosciences GmbH (Germany) and supplied as a 1mM aqueous solutio n. The assay buffer containes 10 mM HEPES pH 7.4 (Applichem), 150 mM NaCl (Sigma), 5 mM MgCl 2 (Sigma), 1 mM DTT (Thermofisher), 0.05% BSA Fraction V, pH 7.0, (ICN Biomedicals), 0.0025% (v/v) Igepal (Sigma). The expression and purification of N‐terminal GST‐tagged hK‐RasG12C and N‐terminal His‐tagged hSOS2 is described below. Concentrations of protein batches used are optimized to be within the linear range of the HTRF signal. A hRas working solution is prepared in assay buffer containing typically 100 nM GST‐hK‐RasG12C and 2 nM antiGST Tb (Cisbio, France). A hSOS2 working solution is prepared in assay buffer containing typically 20nM hSOS2 and 200 nM EDA‐GTP‐Dy647P1. An inhibitor control solution is prepared in assay buffer containing 200 nM EDA‐GTP‐Dy647P1 without hSOS2. Fifty nl of a 100‐fold concentrated solution of the test compound in DMSO are transferred into a black microtiter test plate (384 or 1536, Greiner Bi o‐One, Germany). For this, either a Hummingbird liquid handler (Digilab, MA, USA) or an Echo acousti c system (Labcyte, CA, USA) is used. All steps of the assay are performed at 20°C. A v olume of 2.5 µl of the hRas working solution is a dded to all wells of the test plate using a Multidrop dispenser (Thermo Labsystems). After 10 min preincubation, 2.5 µl of the hSOS2 working solution are added to all wells except for those wells at the side of the test plate that are subsequently filled with 2.5 µl of the inhibitor control solution. After 30 min incubation the fluorescence is measured with a Pherastar (BMG, Germany) using the HTRF module (excitation 337nm, emission 1: 620nm, emi ssion 2: 665nm). The ratiometric data (emission 2 divided by emission 1) are normalized using the controls (DMSO = 0% inhibition, inhibition control wells with inhibitor control solution = 100% inhibition). Compounds are tested in duplicates at up to 11 concentrations (for example 20 µM, 5,7 µM, 1,6 µM, 0,47 µM, 0,13 µM, 38 nM, 11 nM, 3,1 nM, 0,89 nM, 0,25 nM and 0,073 nM). IC50 values are calculated by 4‐ Parameter fitting using a commercial software package (Genedata Screener, Switzerland). EGFR kinase assay EGFR inhibitory activity of compounds of the present invention is quantified employing the TR‐FRET based EGFR assay as described in the following parag raphs.
Epidermal Growth Factor Receptor (EGFR) affinity pu rified from human carcinoma A431 cells (Sigma‐ Aldrich, # E3641) is used as kinase. As substrate for the kinase reaction the biotinylated peptide biotin‐Ahx‐AEEEEYFELVAKKK (C‐terminus in amid form ) is used which can be purchased e.g. form the company Biosyntan GmbH (Berlin‐Buch, Germany). For the assay 50 nL of a 100fold concentrated solution of the test compound in DMSO is pipetted into a black low volume 384well microtiter plate (Gr einer Bio‐One, Frickenhausen, Germany), 2 µL of a solution of EGFR in aqueous assay buffer [50 mM Hepes/HCl pH 7.0, 1 mM MgCl 2 , 5 mM MnCl 2 , 0.5 mM activated sodium ortho‐vanadate, 0.005% (v/v) Tween‐20] are added and the mixture is incubated for 15 min at 22°C to allow pre‐binding of the test compounds to the enzyme before the start of the kinase reaction. Then the kinase reacti on is started by the addition of 3 µL of a solut ion of adenosine‐tri‐phosphate (ATP, 16.7 µM => final conc. in the 5 µL assay volume is 10 µM) and substrate (1.67 µM => final conc. in the 5 µL assay volume is 1 µM) in assay buffer and the re sulting mixture is incubated for a reaction time of 20 min at 22°C. The concentration of EGFR is adjusted depending of the activity of the enzyme lot and is chosen appropriate to have the assay in the linear range, typical concentration are about 3 U/ml. The reaction is stopped by the addition of 5 µl of a solution of HTRF detection reagents (0.1 µM streptavidine‐XL665 [Cis Biointernational] and 1 nM PT66‐Tb‐Cryptate, an terbium‐cryptate labelled anti‐phospho‐tyrosine antibody from Cis Biointernational [instead of the PT66‐Tb‐cryptate PT66‐Eu‐Chelate from Perkin Elmer can also be used]) in an aqueous EDTA‐solution (80 mM EDTA, 0.2 % (w/v) bovine serum albumin in 50 mM HEPES pH 7.5). The resulting mixture is incubated 1 h at 22°C to allow the binding of the biotinylated phosphorylated peptide to the streptavidine‐XL665 and the PT66‐Eu‐Chelate. Subsequently the amount of phosphorylated substrate is evaluated by measurement of the resonance energy transfer from the PT66‐Tb‐Cryptate to the streptavidine‐XL665. There fore, the fluorescence emissions at 620 nm and 665 nm after excitation at 337 nm are measured in a HTRF reader, e.g. a Pherastar (BMG Labtechnologies, Offenburg, Germany) or a Viewlux (Per kin‐Elmer). The ratio of the emissions at 665 nm and at 622 nm is taken as the measure for the amount of phosphorylated substrate. The data are normalised (enzyme reaction without inhibitor = 0 % inhibition, all other assay components but no enzyme = 100 % inhibition). Usually the test compo unds are tested on the same microtiterplate in 11 different concentrations in the range of 20 µM to 0.072 nM (e.g. 20 µM, 5.7 µM, 1.6 µM, 0.47 µ M, 0.13 µM, 38 nM, 11 nM, 3.1 nM, 0.89 nM, 0.25 n M and 0.072 nM, the dilution series are prepared separately before the assay on the level of the 100fold concentrated solutions in DMSO by serial dilutions, the exact concentrations may vary depending on the pipettor used) in duplicate values for each concentration and IC50 values are calculated by a 4 parameter fit.
Cellular assays 3D‐Softagar MiaPaca‐2 (ATCC CRL‐1420) and NCI‐H 1792 (ATCC CRL‐5895) Day 1: Softagar (Select Agar, Invitrogen, 3% in ddH2O autoclaved) is boiled and tempered at 48°C. Medium (MiaPaca‐2: DMEM/Ham's F12; [Biochrom; # FG 4815, with stable Glutamine] 10% FCS and 2.5% Horse Serum, H1792: RPMI 1640; [Biochrom; # FG 1215, with stable Glutamine and 10%FCS]) is tempered to 37°C; Agar (3%) is diluted 1:5 in medium (=0.6%) and 50 µl/well plated into 96 well plates (Corning, #3904), wait at room temperature unt il the agar is solid. 3% agar is diluted to 0.25% in medium (1:12 dilution) and tempered at 42°C. Cells are trypsinized, counted and tempered at 37°C; cells (MiaPaCa‐2: 125‐150, NCI‐H1792: 1000 ) are resuspended in 100 µl 0.25% Agar and plated. Wait at room temperature until the agar is solid. O verlay wells with 50 µl medium. Plate sister wells in separate plate for time zero determination. All p lates are incubated overnight 37°C and 5% CO2. Day 2: Measurement of time zero values: Add 40 µl Cell Titer 96 Aqueous Solution (Promega) per well, (light sensitive) and incubate in the dark at 37°Cand 5% CO2. Absorption is measured at 490 nm and reference wavelength 660 nm. DMSO‐prediluted tes t compounds are added with HP Dispenser to a final DMSO concentration of 0.3%. Day 10: Measurement of test compound and control treated wells with Cell Titer 96 AQueous according to time zero. The IC50 values were determi ned using the four parameter fit. Active RAS in Calu‐1 cells (CLS 300141) 40.000 Calu‐1 cells are seeded in 96well plate (NUNC161093) for 48h at 37°C/5%CO2 (10%FBS (S0615), DMEM/Ham's F‐12 (Biochrom; # FG 4815), 2mM L‐Glutamine). After that, medium is changed to FBS‐free medium and the cells were incu bated for further 24h at 37°C/5%CO2. Cells are treated with varying concentrations of DMSO‐predilute d test compounds (final 0.1%) for 30 min at 37°C/5%CO2. Supernatant with test compounds is discarded and, after that, treated cells are stimulated with 100ng/ml EGF (Sigma#E9644, diluted in serum free medium) for 3 minutes. Cells were treated with lysis buffer and all next steps were performed on ice according to the supplier's manual of G‐LISA Kit (Cytoskeleton BK131, Ras Activ ation Assay). Finally, the content of active Ras is measured by detecting the absorbance at 490 nm (Teca n Sunrise). The value of EGF‐stimulated cells is set as 100%, whereas the value of untreated cell s is set as 0%. The IC50 values were determined using the four parameter fit. Active Ras in Hela cells (ATCC CCL‐2) 30.000 Hela cells are seeded in 96well plate for 96 h at 37°C (10%FBS, DMEM/Ham's F‐12, 2mM L‐ Glutamine). After that, medium is changed in to FBS free medium for 24h. Cells are treated with varying concentrations of test compounds for 30 min. After that, treated cells are stimulated with
100ng/ml EGF for 2 minutes. Cells are treated wit h lysis buffer and all next steps are performed on ice according to the supplier's manual of G‐LISA K it (Cytoskeleton BK131, Ras Activation Assay). Finally, the content of active Ras is measured by d etecting the absorbance at 490 nm. The value of EGF‐stimulated cells is set as 100%, whereas the v alue of untreated cells is set as 0%. The results given as % reflecting the inhibition of formation of active Ras compared to control. The IC50 values are determined using the four parame ter fit. pERK HTRF in MOLM‐13 (DMSZ ACC 554) 10000 MOLM‐13 cells are seeded in HTRF 384well low volume plate (Greiner bio‐one #784080) in medium (RPMI 1640 + 10% FCS). After 24 hours, cells are treated with varying concentrations of test compounds for 1h. Next steps are performed to the s upplier's manual Advanced phospho‐ERK1/2 (#64AERPEH) Cisbio one‐plate assay protocol. The con tent of pERK is measured with PHERAstar HTRF protocol, calculated Ratio*1000. The calculated ratio of DMSO‐treated cells is set as 100% and the calculated ratio of negative control is set as 0% (maximum possible effect). The results given as IC50 reflecting the inhibition of formation of pERK compared to DMSO control and negat ive control and normalized according to cell number. The IC50 values are determined by means of a 4 par ameter fit. pERK HTRF in Calu‐1 (CLS 300141) 5000 Calu‐1 cells are seeded in HTRF 384well low volume plate (Greiner bio‐one #784080) in medium (McCoy's 5A + 10% FCS). After 24 hours, cells are treated with varying concentrations of test compounds for 24h. Next steps are performed to the supplier's manual Advanced phospho‐ERK1/2 (#64AERPEH) Cisbio one‐plate assay protocol. The con tent of pERK is measured with PHERAstar HTRF protocol, calculated Ratio*1000. The calculated ratio of DMSO‐treated cells is set as 100% and the calculated ratio of negative control is set as 0% (maximum possible effect). The results given as IC50 reflecting the inhibition of formation of pERK compared to DMSO control and negat ive control and normalized according to cell number. The IC50 values are determined by means of a 4 par ameter fit. pERK HTRF in K‐562 (ATCC CCL‐243) 10000 K‐562 cells are seeded in HTRF 384well low volume plate (Greiner bio‐one #784075) in medium (RPMI 1640 + 10% FCS) and treated with varyi ng concentrations of test compounds for 1h. Next steps are performed to the supplier's manual Ad vanced phospho‐ERK1/2 (#64AERPEH) Cisbio
one‐plate assay protocol. The content of pERK is measured with PHERAstar HTRF protocol, calculated Ratio*1000. The calculated ratio of DMSO‐treated cells is set as 100% and the calculated ratio of negative control is set as 0% (maximum possible effect). The results given as IC50 reflecting the inhibition of formation of pERK compared to DMSO control and negat ive control and normalized according to cell number. The IC50 values are determined by means of a 4 par ameter fit. pERK assay in NCI‐H358 cells (ATCC CRL‐5807) for combination experiments 5000 NCI‐H358 cells are seeded in HTRF 384well low volume plate (Greiner bio‐one #784080) in medium (RPMI + 10% FCS). After 24h, cells are treat ed for 1h with component A and with component B for single compound treatments (final concentration ranges covering the expected IC50 values), and in nine different fixed‐ratio combinations of compound A (D1) and compound B (D2) (0.9xD1+0.1xD2, 0.8xD1+0.2xD2, 0.7xD1+0.3xD2, 0.6xD1+0.4xD2, 0.5xD1+0.5xD2, 0.4xD1+0.6xD2, 0.3xD1+0.7xD2, 0.2xD1+0.8xD2, 0.1xD1+0.9xD2) using a Te can HP digital dispenser. Next steps are performed to the supplier's manual Advanced phospho‐ERK1/2 (#64AERPEH) Cisbio one‐plate assay protocol. The content of pERK is m easured with PHERAstar HTRF protocol, calculated Ratio*1000. IC50 values (inhibitory concentration at 50% of maximal effect) are determined by means of a 4 parameter fit on measurement data which are normalize d to vehicle (DMSO) treated cells (=100%) and measurement readings taken immediately before comp ound exposure (=0%). IC50 isobolograms are plotted with the actual concentrations of the two compounds on the x‐ and y‐axis, and the combination index (CI) is calculated according to the median‐effect model of Chou‐Talalay (Chou T.C. 2006 Pharmacol. Rev.). A CI of <0.8 is defined a s more than additive (synergistic) interaction, and a CI of >1.2 is defined as antagonistic interaction. P‐EGFR assay (In‐Cell Western) in Hela cells (ATC C CCL‐2) After stimulation with EGF, the EGF receptor autophosphorylates at Y1173. In‐cell Western assay simultaneously detect two targets at 700 and 800nm u sing two spectrally distinct near‐infrared dyes. With a specific antibody, phosphorylated EGFR can be quantified and the samples can be normalized with total EGFR antibody parallel. 25000 Hela cells are seeded in 96well plate (NUNC161 093) for 24 h at 37°C/5%CO2 (10%FBS (S0615), DMEM/Ham's F‐12 (Biochrom; # FG 4815), 2mM L‐Glut amine). After that, medium is changed to FBS‐ free medium and the cells are incubated for further 24h at 37°C/5%CO2.
Cells are treated with varying concentrations of D MSO‐prediluted test compounds (final 0.1%) for 30 minutes and finally with 100ng/ml EGF (Sigma#E9644, d iluted in serum free medium) for 2 minutes. Cells are treated according the manual of EGFR Near Infrared In‐Cell ELISA Kit (Pierce #62210). If not specified, all buffers and antibodies are part of th is kit. Cells are fixed with 4% formaldehyde, washed twice with 100µl per well with TRIS‐buffered saline with Surfact‐Amps 20, permeabilized with 100µl TRIS ‐buffered saline with Surfact‐Amps X‐100, wash again with 100µl TRIS‐buffered saline, and finally 200µl blocking buffer are added for 60 minutes at room temperature. Fixed and washed cells are incubate d with primary antibody mix (P‐EGFR; EGFR) overnight at 2‐8°C. After washing with 100µl TRIS ‐buffered saline with Surfact‐Amps 20, secondary IRDye‐labeled antibody mix (DyLight 800 Goat Anti‐ Rabbit IgG, Pierce SA5‐35571; DyLight 680 Goat Anti‐Mouse IgG, Pierce 35518) is added for 1h at room temperature and washed again. Plates are scanned with LiCor Odyssey Infrared Imager at 800nm for P‐EGFR and at 700nm for total EGFR. The quotient of 800nm and 700nm for EGF only treated ce lls is set as 100% and the quotient of 800nm and 700nm of untreated cells is set as 0%. The IC5 0 values are determined using the four parameter fit. pERK assay in NCI‐H358 cells (ATCC CRL‐5807) for combination experiments NCI‐H358 human non‐small cell lung tumor cells (ATCC CRL‐5807) are propagated in a humidified 37°C incubator in RPMI1640 growth medium (Thermo Fis her Gibco, #61870‐010) supplemented with 10% fetal calf serum (Biochrom, #S 0615). For analys is of combination effects between compound A and compound B, cells are plated in 384‐well plate s (Greiner bio‐one, #784080) at a density of 20,00 0 cells per well in 8 microL of growth medium supplem ented with 10% fetal calf serum. After 24h, cells are treated with component A and with component B for single compound treatments (final concentration ranges covering the expected IC50 values), and in nine different fixed‐ratio combinations of compound A (D1) and compound B (D2) (0.9xD1+0.1xD2, 0.8xD1+0.2xD2, 0.7xD1+0.3xD2, 0.6xD1+0.4xD2, 0.5xD1+0.5xD2, 0.4xD1+0.6xD2, 0.3xD1+0.7xD2, 0.2xD1+0.8xD2, 0.1xD1+0.9xD2) using a Tecan HP digital dispenser. Th e cells are incubated for 60 minutes at 37°C. 4 microL/well of a freshly prepared solution of 0.6 nanog/microL of epid ermal growth factor (Sigma, #E9644) in RPMI1640 medium are added using a Thermo Fisher Multidrop device (final concentration 200 nanog/milliL). The cells are incubated for another 3 minutes immediately followed by the detection of total ERK1/2 and phosphorylated ERK1/2 a t positions Thr202/Tyr204 using commercial HTRF detection kits (Cisbio: total ERK1/2, 64NRKPEG; phospho‐ERK1/2, 64AERPEH) and a PHERAstar microplate reader device (BMG Labtech). Cell lysis and detection are performed according to the manufacturer’s recommendations. The ratio of phosphor ylated ERK1/2 to total ERK1/2 protein are calculated and IC50 values (inhibitory concentration at 50% of maximal effect) are determine d by means of a 4 parameter fit on measurement data whic h are normalized to vehicle (DMSO) treated cells (=100%). IC50 isobolograms are plotted with the actual concentration s of the two compounds on the x‐ and y‐axis, and the combination index (CI) is calculated according to the median‐effect model of Chou‐Talalay (Chou T.C. 2006 Pharmacol. Re v.). A CI of <0.8 is defined as more than addit ive (synergistic) interaction, and a CI of >1.2 is de fined as antagonistic interaction. Table 1: IC 50 values of some examples in the K‐RasG12C – SOS interaction assay, in K‐RasG12C activation by SOS, in K‐Ras activation by SOS high GTP and in K‐Ras‐wt activation by SOS
As exemplified in table 1, the compounds of the present invention inhibit the binding of hSOS1 to hKRAS, which was measured in the biochemical hK‐Ras G12C ‐hSOS1 interaction assay (assay 1). The ability to inhibit the hKRAS‐hSOS1 interaction results in the inhibition of hKRAS activation by the compounds, as measured in biochemical assay 3, which quantifies the hSOS1‐mediated nucleotide exchange from hK‐RasG12C‐GDP to hK‐RasG12C loaded with a fluorescent GTP‐analog. Furthermore, the compounds of the present invention show the abil ity to inhibit the nucleotide exchange reaction catalyzed by hSOS1 in the presence of a high concentration of 50 µM GTP, as measured in assay 2. This ability increases the chance that the compounds will be able to inhibit hSOS1 mediated hKRAS‐ activation inside cells, where high GTP concentrations are present. The chemical structure of the compounds of the present invention is similar to kno wn inhibitors of EGFR‐kinase. As shown in table 1, most compounds are inactive against EGFR‐kinase up to the highest concentration measured in the assay (>20 µM). The assay data of the large number of compounds in table 1 gives evidence that compounds which have a pharmacological profile as tested according to assays 1 to 3 and as described in the preceding paragraph will be generally useful to inhibit hSOS1 mediated hKRAS‐activation inside cells, where high GTP concentrations are present and activity agai nst EGFR‐kinase up to highest concentrations (>20 µM) will not be measured in the assay. Therefore an even further aspect of the present invention refers to the use of a compound which inhibits the binding of hSOS1 to human H‐ or N‐ or K‐RAS including their clinically known mutation s and which inhibits the nucleotide exchange reaction catalyzed by hSOS1 in the presence of a concentration of 20 µM or lower, but which is substantially inactive against EGFR‐kinase at concentrations of 20 µM or lower for the preparation of a medicament for the treatment or prophylaxis of a hyperproliferative disorder. Particularly this aspect refers to the use of a compound which inhibits the binding of hSOS1 specifically to hK‐RasG12C protein and which inhibit s the nucleotide exchange reaction catalyzed by hSOS1 in the presence of a concentration of 20 µM or lower, but which is substantially inactive against EGFR‐kinase at concentrations of 20 µM or lower for the preparation of a medicament for the treatment or prophylaxis of a hyperproliferative disor der. Expression of hK‐RasG12C, hSOS1, hSOS1_12 and hSOS2 in E. coli: The applied DNA expression constructs encoding the following protein sequences and its corresponding DNA sequences were optimized for expression in E. coli and synthesized by the GeneArt Technology at Life Technologies: Human K‐Ras (P01116‐2): hK‐RasG12C (amino acid 1‐169) Human SOS1 (Q07889): hSOS1 (amino acid 564‐1049) hSOS1_12: (amino acid 564‐1049 which is fused at its N‐terminus with the amino acid sequence GAMA Human SOS2 (Q07890): hSOS2 (amino acid 564‐1043) These expressions construct additionally encoded att‐site sequences at the 5´and 3´ ends for subcloning into various destination vectors using the Gateway Technology as well as a TEV (Tobacco Etch Virus) protease site for proteolytic cleavage of tag sequences. The applied destination vectors were: pD‐ECO1 (an in‐house derivate of the pET vector series from Novagen with ampicillin resistance gene) which provides an N‐terminal fusion of a GST‐tag to the integrated gene of interest. pD‐ECO5 (also an in‐house derivative of the pET vector series with ampicillin resistance gene) which provides a N‐terminal fusion of a His10‐tag to t he integrated gene. To generate the final expression vectors the expression construct of hK‐Ras_G12C was cloned into pD‐ECO1. hSOS1, hSOS1_12 as well as hSOS2 were cloned into pD‐ECO5. The resulting expression vectors were termed pD‐ECO1_hK‐ RasG12C, pD‐ECO5_hSOS1, pD‐ECO5_hSOS1_12, pD‐ECO5_h SOS2 Sequences: GST‐hK‐RasG12C (G12C mutation according to numberin g in P01116‐2)
E. coli Expression: The expression vectors were transformed into E. coli strain BL21 (DE3). Cultivation of the transformed strains for expression was done in 10 L and 1 L fermenter. The cultures were grown in Terrific Broth media (MP Biomedicals, Kat. #113045032) with 200 ug/mL ampicillin at a temperature of 37 °C to a density of 0.6 (OD600), shifted to a temperature of 27 °C (for hK‐Ras expression vectors) or 17 °C (for hSOS expression vectors), induced for expression with 100 mM IPTG and further cultivated for 24 hours. Purification After cultivation the transformed E. coli were harvested by centrifugation and the resulting pellet was suspended in a lysis buffer (see below) and lys ed by passing three‐times through a high pressure device (Microfluidics). The lysate was centrifuged (49000g, 45 min, 4 °C) and the supernatant used for further purification. An Äkta chromatography system was used for all furt her chromatography steps. Purification of GST‐hK‐RasG12C for biochemical assa ys E. coli culture (transformed with pD‐ECO1_hK‐RasG12C) from a 10L fermenter was lysed in lysis buffer (50mM Tris HCl 7.5, 500mM NaCl,1mM DTT, 0,5% CHAPS, Complete Protease Inhibitor Cocktail‐(Roche)). As a first chromatography step the centrifuged lysate was incubated with 50mL Glutathione Agarose 4B (Macherey‐Nagel; 745500.100) in a spinner flask (16 h, 10°C). The Glutathione Agarose 4B loaded with protein was transf erred to a chromatography column connected to an Äkta chromatography system. The column was wa shed with wash buffer (50mM Tris HCl 7.5, 500mM NaCl, 1mM DTT) and the bound protein eluted with elution buffer (50mM Tris HCl 7.5, 500mM NaCl, 1mM DTT, 15mM Glutathione). The main fra ctions of the elution peak (monitored by OD280) were pooled. For further purification by size‐exclusion chromatogr aphy the above eluate volume was applied to a column Superdex 200 HR prep grade (GE Healthcare) an d the resulting peak fractions of the eluted fusion protein were collected. The final yield of hK ‐RasG12C was about 50 mg purified fusion protein
per L culture and the final product concentration was about 1 mg/mL. Native mass spectrometry analyses of the final purified K‐RasG12C demonstrate d its homogeneous load with GDP. Purification of His10‐hSOS1 and His10‐hSOS2 for bi ochemical assays E. coli transformed with pD‐ECO5_hSOS1 or pD‐ECO5_hSOS2 were cultured and induced in a fermenter, harvested and lysed in lysis buffer (25mM Tris HCl 7.5, 500mM NaCl, 20mM Imidazol, Complete EDTA‐free (Roche)). For immobilized metal ion affinity chromatography (IMAC) the centrifuged lysate (50 000 xg, 45 min, 4°C) was incubated with 30mL Ni‐NTA (Macherey‐Nagel; #745400.100) in a spinner flask (16 h, 4°C) and subsequently transferred to a chromatography column connected to an Äkta chromatography system. The column was rinsed with wash buffer (25mM Tris HCl 7.5, 500mM NaCl, 20mM Imidazol) and the bound protein eluted with a linear gradient (0‐100%) of elution buffer (25mM Tris HCl 7.5, 500mM NaCl, 300mM Imidazol). The main fractions of the elution peak (monitored by OD280) containing homogenous His10‐hSOS were pooled. The final yield of His10‐hSOS1 was about 1 10 mg purified protein per L culture and the final product concentration was about 2 mg/mL. For His10‐ hSOS2 the final yield was 190 mg per L culture and the product concentration 6 mg/mL. Purification of hSOS1_12 To produce tag‐free hSOS1_12 the same process consi sting of 4 chromatography steps applying an Äkta system was used as decribed here below for hS OS1. His10‐hSOS1_12 was expressed in E. coli transformed with pD‐ECO5_hSOS1_12 as described above. For IMAC the centrifuged lysate was directly applied to a 30 mL (or 50 mL) column with Ni‐NTA (Macherey‐Nagel) in an Äkta system, rinsed with wash buffer (25mM Tris HCl 7.5, 500mM NaCl, 20mM Imidazol) and the bound protein was eluted with a linear gradient (0‐100%) of elution buffer (25mM Tris HCl 7.5, 500mM NaCl, 300mM Imidazol). The main fractions of the elution peak (monitored by OD280) were passed over a HiPrep Desal ting column (GE; #17‐5087‐01) to change to the cleavage buffer (25mM Tris HCl 7.5, 150mM NaCl, 1mM DTT). The adjusted protein solution was treated with purified His‐TEV protease (ratio hSOS1 : TEV, w/w, 30:1) for 16 h at 4 °C and afterwar ds passed over a Ni‐NTA column to remove non‐cleaved hSOS1 protein, cleaved tag and His‐TEV. The pooled flow through fractions with the processed hSOS 1 were concentrated using a Amicon Ultra 15 Ultracel‐10 device (Centrifugal Filter 10000 NMWL; Merck‐Millipore #UFC901024) and applied to size‐exclusion chromatography column with Superdex 200 HR prep grade (GE Healthcare) in SEC buffer (25mM Tris HCl 7.5, 100mM NaCl). The final y ield of tag‐free protein for SOS1_12 was about 245 mg per liter cell culture was. The final product (tag‐free ) concentration for hSOS1_12 was 30.7 mg/mL. Complex formation and Crystallization of hSOS1_12 with SOS1 inhibitors The catalytic domain of human SOS1 (hSOS1) in complex with inhibitors can be crystallized using construct hSOS1_12. It is identical to the construct published by Freedman et al. (Ref. 1). It comprises of hSOS1 residues Glu564 to Thr1049 with an addition al four amino acids (Gly‐Ala‐Met‐Ala) at the N terminus and is shown in Figures X1 and X2 below. For inhibitor‐complex formation, frozen aliquots of the hSOS1_12 protein (concentration 30.7 mg/ml) in buffer (25mM Tris HCl 7.5/50mM NaCl/ 1mM DTT) are thawed and the respective SOS1 inhibitor is added before setting up of the crystallization experiment (co‐crystallization approach) or soaked in to pre‐formed apo crystals (soaking approach). For the co‐crystallization approach, the inhibitor i s added from a 200 mM DMSO stock solution to a final inhibitor concentration of 2 mM and the mix i s incubated over night at 4°C. The complex can be crystallized using the Hanging Drop method. Crystals grow at 20°C. Drops are made from 1 µl hSOS1_12:inhibitor mix, 1 µl reservoir solution (20 30 % % (v/v) ethylenglycole) and 0.2 µl seed stoc k. The seed stock was generated from hSOS1 crystals pre viously obtained in an initial screen using the same hSOS1_12 construct and a reservoir solution of 25% ethylene glycol. For the soaking approach, apo SOS1 crystals (grown using the same procedure as described above, just without addition of an inhibitor) are soaked for 2 to 24 hours with 2 mM ligand. Data Collection and Processing SOS1‐inhibitor crystals are directly shock frozen in liquid nitrogen. Diffraction data sets collected at synchrotrons can be processed using the programs XDS and XDSAPP. Structure determination and refinement The crystal form described here was first obtained a nd solved for a hSOS1_12 crystal grown in the presence of another inhibitor of the same chemical series, from a reservoir solution composed of 25% ethylene glycol. This initial structure was solve d using the Molecular Replacement method with the program PHASER from the CCP4 program suite and the published structure of hSOS1 (PDB entry 2ii0, Ref. 1) as search model. The data sets for f urther SOS1:inhibitor crystal structures can be solved by Molecular Replacement using PHASER and an earlier in‐house SOS1:inhibitor co‐complex structure as starting model. 3D models for the inhibitors are generated using the program Discovery Studio (company Biovia) and parameter files for crystallographic refinement and model building are generated using software PRODRG. The inhibitors can be built manually built into the electron density maps using the program COOT, followed by several cycles of refinement (using program REFMAC as part of the CCP4 program suite) and rebui lding in COOT. Figure X1: Sequence of hSOS1_12 with N‐terminal His tag (His10‐hSOS1_12) before cleavage by TEV protease.