KCC2 potentiators and uses thereof
Patent Information
- Application Number
- PCT/US2026/021104
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2026-02-20
- Filing Date
- 2026-03-27
- Publication Date
- 2026-10-01
Smart Images
Figure IMGF000001_0001 
Figure IMGF000002_0001 
Figure IMGF000002_0002
Abstract
Description
[0001] PATENT
[0002] ATTORNEY-DOCKET NO.: 51478-036WO4
[0003] KCC2 POTENTIATORS AND USES THEREOF
[0004] BACKGROUND OF THE INVENTION
[0005] Potassium chloride cotransporter-2 (KCC2) has been linked to neurological disorders, psychiatric disorders, and central nervous system injuries, and has been linked to neurological functions such as sensory, motor, cognitive, and / or developmental functions in the affected individual. These disorders often result in profound and irreversible neurological effects that pose severe challenges to an afflicted patient’s everyday life. Few therapeutics have been studied or utilized to treat these neurological disorders, which causes severe challenges and suffering for these patients. Additionally, the few that have been studied or utilized are not adequately sufficient to reduce the individual’s suffering or improve recovery from these neurological disorders. Accordingly, there is a need for novel therapeutic agents for the treatment of neurological disorders.
[0006] SUMMARY OF THE INVENTION
[0007] The present disclosure provides compounds, compositions, and methods for treating or preventing neurological disorders in a patient. The disclosed methods include administration to a subject suffering from a neurological disorder of a compound disclosed herein. The disclosure further provides pharmaceutical compositions containing one of the compounds described herein. The disclosure further provides compounds and pharmaceutical compositions for use as a medicament. The disclosure further provides compounds and pharmaceutical compositions for use in the treatment or prevention of neurological disorders.
[0008] In the first aspect, the disclosure provides a compound having the structure of Formula (I):
[0009]
[0010] o
[0011] Formula (I),
[0012] or a pharmaceutically acceptable salt thereof, wherein:
[0013] R1is optionally substituted C5-C12 aryl, optionally substituted C3-C12 heteroaryl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[0014] R2is H, optionally substituted C1-C4 alkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[0015] R3is H, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[0016] R4is H, optionally substituted C1-C5 alkyl, optionally substituted C1-C5 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[0017] R5is optionally substituted C3-C12 heteroaryl, optionally substituted C3-C12 heterocycle, NR6R7, or OR9;
[0018] R6is H or C1-C6alkyl;PATENT
[0019] ATTORNEY-DOCKET NO.: 51478-036WO4
[0020] R7is optionally substituted C1-C6alkyl, C1-C6heteroalkyl, or optionally substituted C3-C12 heterocycle, wherein the alkyl or heterocycle is optionally substituted with 1 or 2 substituents selected from F, OH, C1-C3 alkoxy, and (CH2)nR8;
[0021] R8is 5- or 6-membered heteroaryl, 3- to 5-membered cycloalkyl, or 3- to 6-membered heterocycle;
[0022] R9is C3-C12 heterocycle; and
[0023] n is 1, 2, or 3.
[0024] In some embodiments, R1is optionally substituted C3-C12 cycloalkyl, e.g.,
[0025]
[0026] or. In some embodiments, R1is optionally substituted C5-C12 aryl, wherein the aryl is substituted with one or more groups selected from: halo, CN, optionally substituted C1-C4 alkyl, optionally substituted Ci-04 heteroalkyl, optionally substituted C3-C12 cycloalkyl, optionally substituted C3-C12 heterocycle, SO2Ra, C(O)N(Rb)2, and ORC, wherein the alkyl, heteroalkyl, cycloalkyl, or heterocycle is optionally substituted with one or more groups selected from: halo, C1-C4 alkyl, C3-C12 heterocycle, and C5-C12 aryl;
[0027] Rais C1-C4 alkyl;
[0028] each Rbis, independently, C1-C4 alkyl; and
[0029] Rcis optionally substituted C1-C4 alkyl, optionally substituted C5-C12 aryl, or optionally substituted C3-C12 heteroaryl, wherein the alkyl, aryl, or heteroaryl is optionally substituted with one or more groups selected from: halo and optionally substituted C3-C6 heterocycle, wherein the heterocycle is optionally substituted with one or more halo.
[0030]
[0031] PATENT
[0032] ATTORNEY-DOCKET NO.: 51478-036WO4
[0033]
[0034] In some embodiments, R1is optionally substituted C3-C12 heteroaryl, wherein the heteroaryl is substituted with one or more groups selected from: halo and optionally substituted C1-C4 alkyl, wherein the alkyl is optionally substituted with one or more groups selected from: halo and C1-C4 alkyl.
[0035]
[0036] In some embodiments, R1is optionally substituted C3-C12 heterocycle, wherein the heterocycle is substituted with one or more groups selected from: optionally substituted C1-C4 alkyl and C5-C12 aryl, wherein the alkyl is optionally substituted with one or more groups selected from: OH and oxo.PATENT
[0037] ATTORNEY-DOCKET NO.: 51478-036WO4
[0038] In some embodiments, R1is optionally substituted C3-C12 heterocycle, e
[0039]
[0040] .g., y
[0041]
[0042] In some embodiments, R1is optionally substituted C3-C12 cycloalkyl, e.g.,
[0043]
[0044] In some embodiments, R2is H.
[0045] ^CH
[0046] In some embodiments, R2is optionally substituted C1-C4 alkyl, e.g.,
[0047]
[0048] 3.
[0049] In some embodiments, R3is H.
[0050] In some embodiments, R3is optionally substituted C1-C4 alkyl, wherein the alkyl is substituted with one or more groups selected from: halo, optionally substituted C1-C4 alkyl, and optionally substituted C1-C4 heteroalkyl, wherein the alkyl or heteroalkyl is optionally substituted with one or more halo.
[0051] In some embodiments, R3is optionally substituted C1-C4 alkyl, e.g.,
[0052]
[0053] ^n3,5^, or
[0054]
[0055] In some embodiments, R4is H.
[0056] In some embodiments, R4is optionally substituted C1-C5 alkyl, wherein the alkyl is substituted with one or more groups selected from: halo and optionally substituted C1-C4 alkyl, wherein the alkyl is optionally substituted with one or more halo.
[0057] In some embodiments, R4is optionally substituted C1-C5 alkyl, e.g.,
[0058]
[0059]
[0060] 3
[0061] In some embodiments, R4is optionally substituted C1-C5 heteroalkyl, e.g.,
[0062]
[0063]
[0064] In some embodiments, R4is optionally substituted C3-C12 cycloalkyl, wherein the cycloalkyl is substituted with one or more groups selected from: halo and C1-C4 alkyl.
[0065] In some embodiments, R4is optionally substituted C3-C12 cycloalkyl, e.g.,
[0066]
[0067]
[0068] PATENT
[0069] ATTORNEY-DOCKET NO.: 51478-036WO4
[0070] In some embodiments, R4is optionally substituted C3-C12 heterocycle, wherein the heterocycle is substituted with one or more groups selected from: halo, C1-C4 alkyl, C1-C4 heteroalkyl, and C3-C12 heterocycle, wherein the alkyl is optionally substituted with one or more halo.
[0071]
[0072] In some embodiments, R5is optionally substituted C3-C12 heteroaryl, wherein the heteroaryl is substituted with one or more groups selected from: halo and optionally substituted C1-C4 alkyl, wherein the alkyl is optionally substituted with one or more halo.
[0073]
[0074] In some embodiments, R5is optionally substituted C3-C12 heterocycle, wherein the heterocycle is substituted with one or more groups selected from: halo, OH, CONRdRe, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, and optionally substituted C3-C12 cycloalkyl, wherein the alkyl, heteroalkyl, or cycloalkyl is optionally substituted with one or more halo; and
[0075] each Rdand Reis, independently, C1-C4 alkyl or C5-C12 aryl.
[0076] In some embodiments, R5is optionally substituted C3-C12 heterocycle, e.g.,
[0077]
[0078]
[0079] PATENT
[0080] ATTORNEY-DOCKET NO.: 51478-036WO4
[0081]
[0082] In some embodiments, R6is C1-C6alkyl, e.g., CH3, or
[0083]
[0084] In some embodiments, R5is OR9, e.g.,
[0085] In some embodiments, R9is C3-C12 heterocycle, e.g.,
[0086]
[0087] o'PATENT
[0088] ATTORNEY-DOCKET NO.: 51478-036WO4
[0089] In some embodiments, the compound of Formula (I) has the structure of Formula (l-A):
[0090]
[0091] Formula (l-A),
[0092] or a pharmaceutically acceptable salt thereof.
[0093] In some embodiments, the compound of Formula (I) has the structure of Formula (l-B):
[0094]
[0095] or a pharmaceutically acceptable salt thereof.
[0096] In some embodiments, the compound of Formula (I) has the structure of Formula (l-C):
[0097]
[0098] Formula (l-C),
[0099] or a pharmaceutically acceptable salt thereof.
[0100] In some embodiments, the compound of Formula (I) has the structure of Formula (l-D):
[0101]
[0102] Formula (l-D),
[0103] or a pharmaceutically acceptable salt thereof.PATENT
[0104] ATTORNEY-DOCKET NO.: 51478-036WO4
[0105] In some embodiments, the compound of Formula (I) has the structure of Formula (l-E):
[0106] .0
[0107] N'R2
[0108] , N.. R3
[0109] R5^N^ yN'R4
[0110]
[0111] o
[0112] Formula (l-E),
[0113] or a pharmaceutically acceptable salt thereof.
[0114] In some embodiments, the compound of Formula (I) has the structure of Formula (l-F):
[0115] ^N'R2
[0116] , R:
[0117] R5N %
[0118]
[0119] Formula (l-F),
[0120] or a pharmaceutically acceptable salt thereof.
[0121] In some embodiments, the compound of Formula (I) has the structure of Formula (l-G):
[0122] N'R2
[0123] R5N
[0124]
[0125] I OfR4
[0126] Formula (l-G),
[0127] or a pharmaceutically acceptable salt thereof.
[0128] In some embodiments, the compound of Formula (I) has the structure of Formula (l-H):
[0129] c-.., R'
[0130] R5NN'R.
[0131]
[0132] Formula (l-H),
[0133] or a pharmaceutically acceptable salt thereof.PATENT
[0134] ATTORNEY-DOCKET NO.: 51478-036WO4
[0135] In some embodiments, the compound of Formula (I) has the structure of Formula (l-l):
[0136]
[0137] O
[0138] Formula (l-l),
[0139] or a pharmaceutically acceptable salt thereof.
[0140] In some embodiments, the compound of Formula (I) has the structure of Formula (l-J):
[0141]
[0142] Formula (l-J),
[0143] or a pharmaceutically acceptable salt thereof.
[0144] In some embodiments, the compound of Formula (I) has the structure of Formula (l-K):
[0145]
[0146] Formula (l-K),
[0147] or a pharmaceutically acceptable salt thereof.
[0148] In some embodiments, the compound of Formula (I) has the structure of Formula (l-L):
[0149]
[0150] Formula (l-L),
[0151] or a pharmaceutically acceptable salt thereof.PATENT
[0152] ATTORNEY-DOCKET NO.: 51478-036WO4
[0153] In some embodiments, the compound of Formula (I) has the structure of Formula (l-M):
[0154]
[0155] Formula (l-M),
[0156] or a pharmaceutically acceptable salt thereof.
[0157] In some embodiments, the compound of Formula (I) has the structure of Formula (l-N):
[0158]
[0159] Formula (l-N),
[0160] or a pharmaceutically acceptable salt thereof.
[0161] In some embodiments, the compound of Formula (I) has the structure of Formula (1-0):
[0162]
[0163] Formula (1-0),
[0164] or a pharmaceutically acceptable salt thereof.
[0165] In some embodiments, the compound of Formula (I) has the structure of Formula (l-P):
[0166]
[0167] Formula (l-P),
[0168] or a pharmaceutically acceptable salt thereof.
[0169] In some embodiments, the compound of Formula (I) has the structure of Formula (l-Q):
[0170]
[0171] PATENT
[0172] ATTORNEY-DOCKET NO.: 51478-036WO4
[0173] or a pharmaceutically acceptable salt thereof.
[0174] In some embodiments, the compound of Formula (I) has the structure of Formula (l-R):
[0175]
[0176] Formula (l-R),
[0177] or a pharmaceutically acceptable salt thereof.
[0178] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AL):
[0179]
[0180] or a pharmaceutically acceptable salt thereof.
[0181] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AM):
[0182]
[0183] Formula (l-AM),
[0184] or a pharmaceutically acceptable salt thereof.
[0185] In some embodiments, the compound of Formula (l-AM) has the structure of Formula (l-AM-1):
[0186]
[0187] Formula (l-AM-1),
[0188] or a pharmaceutically acceptable salt thereof.PATENT
[0189] ATTORNEY-DOCKET NO.: 51478-036WO4
[0190] In some embodiments, the compound of Formula (l-AM) has the structure of Formula (l-AM-2):
[0191]
[0192] Formula (l-AM-2),
[0193] or a pharmaceutically acceptable salt thereof.
[0194] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AN):
[0195]
[0196] Formula (l-AN),
[0197] or a pharmaceutically acceptable salt thereof.
[0198] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AU):
[0199]
[0200] O
[0201] Formula (l-AU),
[0202] or a pharmaceutically acceptable salt thereof.
[0203] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AV):
[0204]
[0205] Formula (l-AV),
[0206] or a pharmaceutically acceptable salt thereof.PATENT
[0207] ATTORNEY-DOCKET NO.: 51478-036WO4
[0208] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AW):
[0209] O
[0210] ^N'R2
[0211] 1, R'
[0212] R1%
[0213]
[0214] o
[0215] Formula (l-AW),
[0216] or a pharmaceutically acceptable salt thereof.
[0217] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AX):
[0218] 0
[0219] iL R
[0220] SrR2
[0221] R3
[0222] R5N
[0223]
[0224] O
[0225] Formula (l-AX),
[0226] or a pharmaceutically acceptable salt thereof.
[0227] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AY):
[0228] CN
[0229] N'R2
[0230] R1
[0231] R5N R'
[0232]
[0233] O
[0234] Formula (l-AY),
[0235] or a pharmaceutically acceptable salt thereof.
[0236] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AZ):
[0237] IL KI
[0238] R5N y 'R'
[0239]
[0240] o
[0241] Formula (l-AZ),
[0242] or a pharmaceutically acceptable salt thereof.PATENT
[0243] ATTORNEY-DOCKET NO.: 51478-036WO4
[0244] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BA):
[0245]
[0246] O
[0247] Formula (l-BA),
[0248] or a pharmaceutically acceptable salt thereof.
[0249] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BB):
[0250]
[0251] Formula (l-BB),
[0252] or a pharmaceutically acceptable salt thereof.
[0253] In some embodiments, the compound of Formula (l-BB) has the structure of Formula (l-BB-1):
[0254]
[0255] Formula (l-BB-1),
[0256] or a pharmaceutically acceptable salt thereof.
[0257] In some embodiments, the compound of Formula (l-BB) has the structure of Formula (l-BB-2):
[0258]
[0259] Formula (l-BB-2),
[0260] or a pharmaceutically acceptable salt thereof.PATENT
[0261] ATTORNEY-DOCKET NO.: 51478-036WO4
[0262] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BC):
[0263] ^N'R2
[0264] R!
[0265]
[0266] Y ON'R‘
[0267] Formula (l-BC),
[0268] or a pharmaceutically acceptable salt thereof.
[0269] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BD):
[0270] I
[0271] N. JN
[0272] ^N'R2
[0273] RN'R‘
[0274]
[0275] 0
[0276] Formula (l-BD),
[0277] or a pharmaceutically acceptable salt thereof.
[0278] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BE):
[0279] I
[0280] N. JN
[0281] ^N'R2
[0282] , R:
[0283] R5NN'R‘
[0284]
[0285] Formula (l-BE),
[0286] or a pharmaceutically acceptable salt thereof.
[0287] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BF):
[0288] N
[0289] N'R2
[0290] X. N. _ R:3
[0291] R5NN'R-
[0292]
[0293] Formula (l-BF),
[0294] or a pharmaceutically acceptable salt thereof.PATENT
[0295] ATTORNEY-DOCKET NO.: 51478-036WO4
[0296] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BG):
[0297]
[0298] Formula (l-BG),
[0299] or a pharmaceutically acceptable salt thereof.
[0300] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BH):
[0301]
[0302] Formula (l-BH),
[0303] or a pharmaceutically acceptable salt thereof.
[0304] In some embodiments, the compound of Formula (I) has the structure of Formula (l-S):
[0305]
[0306] O
[0307] Formula (l-S),
[0308] or a pharmaceutically acceptable salt thereof.
[0309] In some embodiments, the compound of Formula (I) has the structure of Formula (l-T):
[0310]
[0311] O
[0312] Formula (l-T),
[0313] or a pharmaceutically acceptable salt thereof.PATENT
[0314] ATTORNEY-DOCKET NO.: 51478-036WO4
[0315] In some embodiments, the compound of Formula (I) has the structure of Formula (l-U):
[0316]
[0317] or a pharmaceutically acceptable salt thereof.
[0318] In some embodiments, the compound of Formula (I) has the structure of Formula (l-V):
[0319]
[0320] Formula (l-V),
[0321] or a pharmaceutically acceptable salt thereof.
[0322] In some embodiments, the compound of Formula (I) has the structure of Formula (l-W):
[0323]
[0324] Formula (l-W),
[0325] or a pharmaceutically acceptable salt thereof.
[0326] In some embodiments, the compound of Formula (I) has the structure of Formula (l-X):
[0327]
[0328] Formula (l-X),
[0329] or a pharmaceutically acceptable salt thereof.
[0330] In some embodiments, the compound of Formula (I) has the structure of Formula (l-Y):
[0331]
[0332] PATENT
[0333] ATTORNEY-DOCKET NO.: 51478-036WO4
[0334] or a pharmaceutically acceptable salt thereof.
[0335] In some embodiments, the compound of Formula (I) has the structure of Formula (I-BI):
[0336]
[0337] Formula (l-BI),
[0338] or a pharmaceutically acceptable salt thereof.
[0339] In some embodiments, the compound of Formula (I) has the structure of Formula (I-BJ):
[0340]
[0341] Formula (l-BJ),
[0342] or a pharmaceutically acceptable salt thereof.
[0343] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BK):
[0344]
[0345] Formula (l-BK),
[0346] or a pharmaceutically acceptable salt thereof.
[0347] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BL):
[0348]
[0349] Formula (l-BL),
[0350] or a pharmaceutically acceptable salt thereof.
[0351] In some embodiments, the compound of Formula (I) has the structure of Formula (l-Z):
[0352]
[0353] Formula (l-Z),
[0354] or a pharmaceutically acceptable salt thereof.PATENT
[0355] ATTORNEY-DOCKET NO.: 51478-036WO4
[0356] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AA):
[0357] R1N-R2
[0358]
[0359] Formula (l-AA),
[0360] or a pharmaceutically acceptable salt thereof.
[0361] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BM):
[0362]
[0363] o
[0364] Formula (l-BM),
[0365] or a pharmaceutically acceptable salt thereof.
[0366] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BN):
[0367] R!N-R2
[0368]
[0369] Formula (l-BN),
[0370] or a pharmaceutically acceptable salt thereof.
[0371] In some embodiments, the compound of Formula (l-BN) has the structure of Formula (l-BN-1):
[0372]
[0373] Formula (l-BN-1),
[0374] or a pharmaceutically acceptable salt thereof.
[0375] In some embodiments, the compound of Formula (l-BN) has the structure of Formula (l-BN-2):
[0376] R1N-R2
[0377] R»SJIYNVCF3
[0378]
[0379] o =
[0380] Formula (l-BN-2),
[0381] or a pharmaceutically acceptable salt thereof.PATENT
[0382] ATTORNEY-DOCKET NO.: 51478-036WO4
[0383] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AB):
[0384] R2-N-R1
[0385]
[0386] Formula (l-AB),
[0387] or a pharmaceutically acceptable salt thereof.
[0388] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AO):
[0389]
[0390] o
[0391] Formula (l-AO),
[0392] or a pharmaceutically acceptable salt thereof.
[0393] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AP):
[0394]
[0395] Formula (l-AP),
[0396] or a pharmaceutically acceptable salt thereof.
[0397] In some embodiments, the compound of Formula (l-AP) has the structure of Formula (l-AP-1):
[0398]
[0399] Formula (l-AP-1),
[0400] or a pharmaceutically acceptable salt thereof.
[0401] In some embodiments, the compound of Formula (l-AP) has the structure of Formula (l-AP-2):
[0402]
[0403] Formula (l-AP-2),
[0404] or a pharmaceutically acceptable salt thereof.PATENT
[0405] ATTORNEY-DOCKET NO.: 51478-036WO4
[0406] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BO):
[0407]
[0408] Formula (l-BO),
[0409] or a pharmaceutically acceptable salt thereof.
[0410] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BP):
[0411]
[0412] Formula (l-BP),
[0413] or a pharmaceutically acceptable salt thereof.
[0414] In some embodiments, the compound of Formula (l-BP) has the structure of Formula (l-BP-1):
[0415]
[0416] Formula (l-BP-1),
[0417] or a pharmaceutically acceptable salt thereof.
[0418] In some embodiments, the compound of Formula (l-BP) has the structure of Formula (l-BP-2):
[0419]
[0420] Formula (l-BP-2),
[0421] or a pharmaceutically acceptable salt thereof.
[0422] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BQ):
[0423] R2N-R1
[0424]
[0425] Formula (l-BQ),
[0426] or a pharmaceutically acceptable salt thereof.PATENT
[0427] ATTORNEY-DOCKET NO.: 51478-036WO4
[0428] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BR):
[0429]
[0430] Formula (l-BR),
[0431] or a pharmaceutically acceptable salt thereof.
[0432] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AC):
[0433]
[0434] Formula (l-AC),
[0435] or a pharmaceutically acceptable salt thereof.
[0436] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AD):
[0437]
[0438] Formula (l-AD),
[0439] or a pharmaceutically acceptable salt thereof.
[0440] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AE):
[0441]
[0442] Formula (l-AE),
[0443] or a pharmaceutically acceptable salt thereof.
[0444] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AQ):
[0445]
[0446] Formula (l-AQ),
[0447] or a pharmaceutically acceptable salt thereof.PATENT
[0448] ATTORNEY-DOCKET NO.: 51478-036WO4
[0449] In some embodiments, the compound of Formula (l-AQ) has the structure of Formula (l-AQ-1):
[0450]
[0451] Formula (l-AQ-1),
[0452] or a pharmaceutically acceptable salt thereof.
[0453] In some embodiments, the compound of Formula (l-AQ) has the structure of Formula (l-AQ-2):
[0454]
[0455] Formula (l-AQ-2),
[0456] or a pharmaceutically acceptable salt thereof.
[0457] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BS):
[0458]
[0459] Formula (l-BS),
[0460] or a pharmaceutically acceptable salt thereof.
[0461] In some embodiments, the compound of Formula (l-BS) has the structure of Formula (l-BS-1):
[0462]
[0463] Formula (l-BS-1),
[0464] or a pharmaceutically acceptable salt thereof.
[0465] In some embodiments, the compound of Formula (l-BS) has the structure of Formula (l-BS-2):
[0466]
[0467] Formula (l-BS-2),
[0468] or a pharmaceutically acceptable salt thereof.PATENT
[0469] ATTORNEY-DOCKET NO.: 51478-036WO4
[0470] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BT):
[0471]
[0472] Formula (l-BT),
[0473] or a pharmaceutically acceptable salt thereof.
[0474] In some embodiments, the compound of Formula (l-BT) has the structure of Formula (l-BT-1):
[0475]
[0476] Formula (l-BT-1),
[0477] or a pharmaceutically acceptable salt thereof.
[0478] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BU):
[0479]
[0480] Formula (l-BU),
[0481] or a pharmaceutically acceptable salt thereof.
[0482] In some embodiments, the compound of Formula (l-BU) has the structure of Formula (l-BU-1):
[0483]
[0484] Formula (l-BU-1),
[0485] or a pharmaceutically acceptable salt thereof.
[0486] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BV):
[0487]
[0488] Formula (l-BV),
[0489] or a pharmaceutically acceptable salt thereof.PATENT
[0490] ATTORNEY-DOCKET NO.: 51478-036WO4
[0491] In some embodiments, the compound of Formula (l-BV) has the structure of Formula (l-BV-1):
[0492]
[0493] Formula (l-BV-1),
[0494] or a pharmaceutically acceptable salt thereof.
[0495] In some embodiments, the compound of Formula (l-BV) has the structure of Formula (l-BV-2):
[0496]
[0497] Formula (l-BV-2),
[0498] or a pharmaceutically acceptable salt thereof.
[0499] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BW):
[0500]
[0501] Formula (l-BW),
[0502] or a pharmaceutically acceptable salt thereof.
[0503] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BX):
[0504]
[0505] Formula (l-BX),
[0506] or a pharmaceutically acceptable salt thereof.
[0507] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BY):
[0508] R\.'R2
[0509]
[0510] Formula (l-BY),
[0511] or a pharmaceutically acceptable salt thereof.PATENT
[0512] ATTORNEY-DOCKET NO.: 51478-036WO4
[0513] In some embodiments, the compound of Formula (I) has the structure of Formula (l-BZ):
[0514] F
[0515]
[0516] F
[0517] Formula (l-BZ),
[0518] or a pharmaceutically acceptable salt thereof.
[0519] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AF):
[0520]
[0521] Formula (l-AF),
[0522] or a pharmaceutically acceptable salt thereof.
[0523] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AR):
[0524] R1'N'R2
[0525] R3
[0526] R4
[0527]
[0528] Formula (l-AR),
[0529] or a pharmaceutically acceptable salt thereof.
[0530] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AG):
[0531]
[0532] Formula (l-AG),
[0533] or a pharmaceutically acceptable salt thereof.
[0534] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AH):
[0535]
[0536] Formula (l-AH),
[0537] or a pharmaceutically acceptable salt thereof.PATENT
[0538] ATTORNEY-DOCKET NO.: 51478-036WO4
[0539] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AS):
[0540]
[0541] Formula (l-AS),
[0542] or a pharmaceutically acceptable salt thereof.
[0543] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AT):
[0544]
[0545] Formula (l-AT),
[0546] or a pharmaceutically acceptable salt thereof.
[0547] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AI):
[0548]
[0549] Formula (l-AI),
[0550] or a pharmaceutically acceptable salt thereof.
[0551] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AJ):
[0552]
[0553] Formula (l-AJ),
[0554] or a pharmaceutically acceptable salt thereof.
[0555] In some embodiments, the compound of Formula (I) has the structure of Formula (l-AK):
[0556]
[0557] Formula (l-AK),
[0558] or a pharmaceutically acceptable salt thereof.
[0559] In some embodiments, the present disclosure provides a compound of Table 1 or a pharmaceutically acceptable salt thereof.PATENT
[0560] ATTORNEY-DOCKET NO.: 51478-036WO4
[0561] In some embodiments, the compound of Formula (I) has the structure:
[0562]
[0563] Formula (ll-J),
[0564] Formula (ll-K), O Formula (ll-L),
[0565] Formula (ll-N),
[0566]
[0567] Formula (ll-P),PATENT
[0568] ATTORNEY-DOCKET NO.: 51478-036WO4
[0569] Formula (ll-U), Formula (ll-V),
[0570]
[0571] PATENT
[0572] ATTORNEY-DOCKET NO.: 51478-036WO4
[0573] Formula (ll-AL),
[0574] Formula (ll-AM-1),
[0575]
[0576] Formula (ll-AP),
[0577]
[0578] Formula (ll-AP-2),PATENT
[0579] ATTORNEY-DOCKET NO.: 51478-036WO4
[0580] Formula (II-AQ), Formula (II-AQ-1),
[0581]
[0582] PATENT
[0583] ATTORNEY-DOCKET NO.: 51478-036WO4
[0584] Formula (II-BB), Formula (II-BB-2),
[0585]
[0586] R4Formula (II-BH),
[0587]
[0588] Formula (II-BJ),PATENT
[0589] ATTORNEY-DOCKET NO.: 51478-036WO4
[0590]
[0591] PATENT
[0592] ATTORNEY-DOCKET NO.: 51478-036WO4
[0593] Formula (II-BU-1),
[0594]
[0595] Formula (II-BV-1), Formula (II-BV-2),
[0596]
[0597]
[0598] Formula (II-BZ), or a pharmaceutically acceptable salt thereof.
[0599] In some embodiments, the present disclosure provides a compound of Table 1 or a pharmaceutically acceptable salt thereof.
[0600] Table 1: Compounds of the Invention
[0601]
[0602]
[0603] PATENT
[0604] ATTORNEY-DOCKET NO.: 51478-036WO4
[0605] Compound
[0606] Structure
[0607] #
[0608] 1
[0609] ” I l
[0610] NH
[0611] 3 ) A ('z z < N ■ '-i
[0612] X X
[0613] I
[0614] 0Z' ZNfl 1
[0615] . J VZ- 011
[0616] { o z ---■■■
[0617] Ao.
[0618] 'N 4
[0619] 4
[0620] O’
[0621] A
[0622] < N N! T?
[0623] 0, „> 0
[0624] ( 2:—
[0625] 'NH
[0626] 5 \°.- " A xNs.
[0627] ’ I
[0628] N V
[0629] O '
[0630] 'NH
[0631] 6 A ■t'I
[0632] AV
[0633] 01
[0634] ' CJ
[0635] 7
[0636] A " A
[0637] o„ >
[0638] I
[0639]
[0640]
[0641] PATENT
[0642] ATTORNEY-DOCKET NO.: 51478-036WO4 Compound
[0643] Structure
[0644] #
[0645] 13
[0646] 1
[0647] 14
[0648] >oZ
[0649] ooo-- ' d- 15
[0650] i0
[0651] 16
[0652] I id
[0653] ° rxZJ Z0rd
[0654] 17
[0655] (k ^JAAy 01^ 1
[0656]
[0657]
[0658] PATENT
[0659] ATTORNEY-DOCKET NO.: 51478-036WO4
[0660]
[0661]
[0662] PATENT
[0663] ATTORNEY-DOCKET NO.: 51478-036WO4
[0664] Compound Compound
[0665] Structure Structure # # /
[0666] Y\o / =o z:o Z=CV / ”z~ ~ YYY
[0667] / Y (Y Z 2 Lt- - T x ACPY; HP H 7\NZH33 38 NY-Y
[0668] \ 0 P Y Y O M
[0669] L -- ks^ O^ J 0 ' O. J 0 ’ 1 “i
[0670] HO. X. 1
[0671] 34 39
[0672] < YY JL Y JM. >
[0673] 6^J o 1
[0674] Y
[0675] yY
[0676] rfP * OT - \ y
[0677] P O
[0678] 35 40 fO - o
[0679] ^ Zv / \ X _
[0680] Y L
[0681] 36 41
[0682] Y
[0683] < N' N' _X y' N "'>
[0684] (k J o •
[0685] 1
[0686] ^QP^P[MH
[0687] 37NYA 42
[0688] J. J 0 1
[0689] lr
[0690]
[0691]
[0692] PATENT
[0693] ATTORNEY-DOCKET NO.: 51478-036WO4
[0694] Compound Compound
[0695] Structure Structure # #
[0696] A / F
[0697] d
[0698] 43 o — >.
[0699] U K W r -y^
[0700] A- AXHX\ y s z / \XX<N
[0701] J
[0702] £ — I A
[0703] 0^ X1o '
[0704] Qz==\ o 2A _
[0705] CO- 44
[0706] 0 '
[0707] f
[0708] m
[0709] J--, Ak
[0710] 45NX
[0711] ^ N xXN, „Xy xhk A
[0712] XX 0 '
[0713] 46
[0714] X.
[0715] 47
[0716]
[0717]
[0718] PATENT
[0719] ATTORNEY-DOCKET NO.: 51478-036WO4
[0720]
[0721]
[0722] PATENT
[0723] ATTORNEY-DOCKET NO.: 51478-036WO4 Compound
[0724] Structure
[0725] #
[0726] \ m _
[0727] 63
[0728] N;:y
[0729] JX / \ I_ JL /
[0730] ® O
[0731] <
[0732] 64
[0733] )
[0734] \ Z0yo z^=\_ r
[0735] / \ / \ X X
[0736] (\z—
[0737] / \. M“
[0738] y_ /
[0739] 65
[0740] o\ ' r ~~
[0741] x X °X
[0742] l l
[0743] 66
[0744] O'x / Z 0
[0745] ^0
[0746] I
[0747] 67
[0748]
[0749]
[0750] PATENT
[0751] ATTORNEY-DOCKET NO.: 51478-036WO4
[0752]
[0753]
[0754] PATENT
[0755] ATTORNEY-DOCKET NO.: 51478-036WO4
[0756]
[0757]
[0758] PATENT
[0759] ATTORNEY-DOCKET NO.: 51478-036WO4
[0760] Compound Compound
[0761] Structure Structure # #
[0762] p p Yo / " F3C^Yr T T 7\— Y y / ll A= A (l==-z~ 93 Y 98 lx AL ( oQ - — D P -— \ K <y- ~ fAA - O^J 0 I 0 J 0
[0763] 1
[0764] F
[0765] F1 1
[0766] 94 99
[0767] n
[0768] H 0!
[0769] F n
[0770] ) O— -., Yr O “y /
[0771] lx A 1O7- 95 fn 100 y Y2
[0772] yi fv j Y zz- / \ T Z ‘ - V
[0773] & / so
[0774] 96 101
[0775] Oxl0
[0776] Y
[0777] 97 102
[0778] rlXX
[0779] 1
[0780]
[0781]
[0782] PATENT
[0783] ATTORNEY-DOCKET NO.: 51478-036WO4 Compound
[0784] Structure
[0785] #
[0786] 105
[0787]
[0788]
[0789] PATENT
[0790] ATTORNEY-DOCKET NO.: 51478-036WO4
[0791] Compound Compound
[0792] Structure Structure # #
[0793] F- EA\ A / F
[0794] 113 " ‘'MH F AA / \ xM ^ A
[0795] 118 AA^HF[? ANA' r Y 4 oFA^ > ^-^A^YNy^.o01— / 1
[0796] AzF
[0797] A-AlW F 114 119
[0798] OAVi Y O x "5c oi A AaL. i
[0799] 115 NA / ^AF 120
[0800] ^NAY >
[0801] O^J 0 '
[0802] F'CV
[0803] F AAX1'---NHp 116 121 N^A^^A F | / v\r ytx^AIAY^ o^ J 0
[0804] iaf F
[0805] r 'Alji Al J
[0806] 117 x*A 122
[0807] A^A
[0808] O-^J o f i
[0809]
[0810]
[0811] PATENT
[0812] ATTORNEY-DOCKET NO.: 51478-036WO4 Compound
[0813] Structure
[0814] #
[0815] 123
[0816] 124
[0817] 125
[0818] 126
[0819] 127
[0820]
[0821]
[0822] PATENT
[0823] ATTORNEY-DOCKET NO.: 51478-036WO4 Compound
[0824] Structure
[0825] #
[0826] O — y \
[0827] 133 \ / \z=__
[0828] / A z z —
[0829] < o z=
[0830] ~n
[0831] 134
[0832] yx_ ~
[0833] G z — X
[0834] 135 J '
[0835] 136 TV^
[0836] (X rWj A 0 •'i ■"’r
[0837] f
[0838] 137
[0839] O s
[0840]
[0841] A
[0842]
[0843] PATENT
[0844] ATTORNEY-DOCKET NO.: 51478-036WO4
[0845]
[0846]
[0847] PATENT
[0848] ATTORNEY-DOCKET NO.: 51478-036WO4
[0849] Compound
[0850] Structure
[0851] #
[0852] "fl
[0853] V
[0854] 153
[0855] AA s zz_
[0856] V / / \ / X
[0857] >5 °
[0858] 154
[0859] 0
[0860] O ±= \
[0861] / X
[0862] A
[0863] ] \ / S V \'
[0864] yyg rj
[0865] 155 LL. j r
[0866] N^J 01
[0867] CF3
[0868] 156
[0869] F,c’'Vi
[0870] 157
[0871] rrk"'V']
[0872] CF,
[0873]
[0874]
[0875] PATENT
[0876] ATTORNEY-DOCKET NO.: 51478-036WO4
[0877]
[0878]
[0879] PATENT
[0880] ATTORNEY-DOCKET NO.: 51478-036WO4
[0881]
[0882]
[0883] PATENT
[0884] ATTORNEY-DOCKET NO.: 51478-036WO4 Compound
[0885] Structure
[0886] #
[0887] Jn - 183 jx r x
[0888] / / v
[0889] / \ J: _
[0890] Q
[0891] F3C^. *n w
[0892] 184
[0893] o^4" o k
[0894] / \ □z^
[0895] < z z —
[0896] _27 / * ^
[0897] / \ \s
[0898] 185 oJ ( o ' — P
[0899] u.
[0900] ^c <\
[0901] T 1
[0902] 4 N
[0903] 186 r iA ^N^NPYN^CF3
[0904] Ck J o
[0905] Y
[0906] 187
[0907]
[0908]
[0909] PATENT
[0910] ATTORNEY-DOCKET NO.: 51478-036WO4
[0911] Compound Compound
[0912] Structure Structure
[0913] # #
[0914] 193 / -A 198
[0915] "i <x J 0v1
[0916] Sx-yiH
[0917] 194 199
[0918] y'xA'AyN- o^J °
[0919] X
[0920] f\ r / — ToX z r ) X °'A o
[0921] =u 1
[0922] z / — z d. z —
[0923] 195 C 2 o - — \» P A QA-*X PP'-NHNAA 200
[0924] yNAAyN^
[0925] c\ fJ o
[0926] cry
[0927] PA-MH
[0928] 196 201
[0929] AyA O'^V'v 1
[0930] 197 202 NA / K yAyy; O^ J 0viR
[0931]
[0932]
[0933] PATENT
[0934] ATTORNEY-DOCKET NO.: 51478-036WO4
[0935]
[0936]
[0937] In some embodiments, the compound is one of the compounds 1-9 of Table 1, or a pharmaceutically acceptable salt thereof.
[0938] In some embodiments, the compound is one of the compounds 1-39 of Table 1, or a pharmaceutically acceptable salt thereof.
[0939] In some embodiments, the compound is one of the compounds 1 -159 of Table 1, or a pharmaceutically acceptable salt thereof.PATENT
[0940] ATTORNEY-DOCKET NO.: 51478-036WO4
[0941] In some embodiments, the compound is one of the compounds 1-210 of Table 1, or a pharmaceutically acceptable salt thereof.
[0942] In another aspect, the disclosure provides a pharmaceutical composition including a compound described herein (e.g., any one of the compounds of Formula (I), Formulas (l-A) to (II-E3Z), and Table 1), ora pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
[0943] In another aspect, the disclosure provides a compound described herein (e.g., any one of the compounds of Formula (I), Formulas (l-A) to (ll-BZ), and Table 1), or a pharmaceutically acceptable salt thereof, ora pharmaceutical composition including said compound or salt thereof and a pharmaceutically acceptable excipient, for use as a medicament.
[0944] In another aspect, the disclosure provides a method for treating or preventing a neurological disorder, which includes administering to a subject in need thereof a therapeutically effective amount of a compound described herein (e.g., any one of the compounds of Formula (I), Formulas (l-A) to (ll-BZ), and Table 1), ora pharmaceutically acceptable salt thereof. In some embodiments, the disclosure provides a method for treating a neurological disorder, which includes administering to a subject in need thereof a therapeutically effective amount of a compound described herein (e.g., any one of the compounds of Formula (I), Formulas (l-A) to (ll-BZ), and Table 1), or a pharmaceutically acceptable salt thereof.
[0945] In some embodiments, the neurological disorder is a neurotraumatic disorder, a neurodevelopmental disorder, a neurodegenerative disorder, neurological pain, epilepsy, or an affective disorder.
[0946] In some embodiments, the neurological disorder is dementia.
[0947] In some embodiments, the dementia is Lewy body dementia.
[0948] In some embodiments, the neurological disorder is dementia induced psychosis.
[0949] In some embodiments, the neurological disorder is epilepsy.
[0950] In some embodiments, the epilepsy is refractory epilepsy, developmental and epileptic encephalopathy (DEE), Lennox-Gastaut syndrome, neurotrauma associated epilepsy (ischemia, stroke, traumatic brain injury), status epilepticus, tumor associated epilepsy, or hypoxic-ischemic encephalopathy.
[0951] In some embodiments, the neurological disorder is spinal cord injury, traumatic brain injury, stroke, peripheral nerve injury, ischemia, myelopathy, hypoxic-ischemic encephalopathy, tumor-associated epilepsy, spasticity, neurological pain, neurotraumatic injury, neurodegenerative disease, or peripheral neuropathy.
[0952] In another aspect, the disclosure provides a method for treating or preventing pain, which includes administering to a subject in need thereof a therapeutically effective amount of a compound described herein (e.g., any one of the compounds of Formula (I), Formulas (l-A) to (ll-BZ), and Table 1), or a pharmaceutically acceptable salt thereof.
[0953] In some embodiments, the pain is neurological pain.
[0954] In some embodiments, the neurological pain is a neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia, or neuralgia.
[0955] In some embodiments, the neurotraumatic injury is traumatic brain injury, stroke, nerve injury or spinal cord injury.PATENT
[0956] ATTORNEY-DOCKET NO.: 51478-036WO4
[0957] In some embodiments, the neurodegenerative disorder is multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia.
[0958] In some embodiments, the neurodevelopmental disorder is autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1, 22q11.2 microdeletion syndrome, or cerebral palsy.
[0959] In some embodiments, the affective disorder is schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder.
[0960] DEFINITIONS
[0961] To facilitate the understanding of the present disclosure, a number of terms are defined below. Terms defined herein have meanings as commonly understood by a person of ordinary skill in the areas relevant to the disclosure. Terms such as “a,” “an,” and “the” are not intended to refer to only a singular entity but include the general class of which a specific example may be used for illustration. The terms “comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps. The terminology herein is used to describe specific embodiments of the disclosure, but their usage does not limit the disclosure, except as outlined in the claims.
[0962] As used herein, the term “about” is used to indicate that a value includes the standard deviation of error for the method being employed to determine the value. In certain embodiments, the term “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of a stated value, unless otherwise stated or otherwise evident from the context (e.g., where such number would exceed 100% of a possible value).
[0963] As used herein, any values provided in a range of values include both the upper and lower bounds, and any values contained within the upper and lower bounds.
[0964] As used herein, the terms “administer” and “administering” are used to indicate the process of providing a therapeutic, pharmaceutical, housing compartment, medication, or the like thereof to a subject. In some embodiments, a pharmaceutical is provided via oral administration.
[0965] As used herein, the term “chloride transporter activity” refers to the transport of chloride, or any surrogate ion, across the plasma membrane. Such transport activity may be measured by direct or indirect means using various methods known in the art, examples of which are described herein.
[0966] As used herein, the term “KCC2 activity” refers to any detectable phenotype associated with KCC2. In an embodiment, KCC2 activity includes, but is not limited to potassium transport, chloride transport, or transport of any surrogate ion which may, for example, be determined by assessing levels (either directly or indirectly) of potassium, chloride, and / or surrogate ion inside and / or outside the cell using, for example, reversal potential measurements with patch clamping methods, chloride / potassium sensitive dyes (see for example Haugland, R. P., Handbook of Fluorescent Probes and Research Products, ninth ed., 2002, Molecular Probes, Inc., Eugene, Oreg., USA) electrodes, etc. In addition, KCC2 activity may also affect the neural cell's anion reversal potential (Eanion). The anion reversalPATENT
[0967] ATTORNEY-DOCKET NO.: 51478-036WO4
[0968] potential may be determined, for example, by using gramicidin-perforated patch clamp recording. In an embodiment, KCC2 activity also includes KCC2-mediated synaptic and / or neurite changes.
[0969] As used herein, the terms “improve” and “improving,” in reference to recovery from a disease or condition, e.g., a neurological disorder, refer to an enhancement of recovery in one or more parameters measuring or quantifying the severity of the neurological disorder relative to the recovery in these parameters in or prior to treatment with the compounds or compositions described herein. Alternatively, improvement may be measured with respect to a reference subject having the same diagnosis as the subject butthat did not receive treatment with a compound or composition of the disclosure. For neurological disorders, such parameters may include motor and sensory function in a subject. Methods for assessing motor and sensory function in a subject suffering from a neurological disorder are known in the art and are further described herein.
[0970] As used herein, the term “level” refers to a change in the measure of a compound, metabolite, or protein activity (e.g., chloride transporter activity or KCC2 activity), or a property thereof, as compared to a reference. The reference can be any useful reference, as defined herein. By a “decreased level” or an “increased level” of a compound or protein activity is meant a decrease or increase in level, as compared to a reference (e.g., a decrease or an increase by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; a decrease or an increase of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; a decrease or an increase by about 0.01 -fold, about 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, about 0.9-fold, or less; or an increase by more than about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5- fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50- fold, about 100-fold, or more). A level of a compound may be expressed in mass / vol (e.g., g / dl, mg / ml, pg / ml, ng / ml), concentration or molarity (e.g., M, mM, pM, nM, pM), or percentage relative to total compound in a sample, or by any other suitable units of measure as described by the disclosure. For example, “increasing levels of KCC2 activity” may be considered a subset of the phrase “potentiating KCC2 activity,” when describing an increase in activity of the potassium chloride cotransporter-2, KCC2, in comparison to a reference level.
[0971] As used herein, the term “neurological disorder” refers to any damage or dysfunction of one or more nerves in a subject. A neurological disorder may include any damage or dysfunction that prevents and / or inhibits one or more electrical and / or chemical transmissions of a sensory and / or motor function signal. A neurological disorder may include any damage or dysfunction that results in a transmission of one or more electrical and / or chemical transmissions of a nerve cell uncontrollably by the subject. A neurological disorder may include damage or dysfunction of one or more nerves located within the central nervous system and / or peripheral nervous system of a subject. A neurological disorder may include damage or dysfunction of a somatic, autonomic, and / or enteric nervous system of a subject. A neurological disorder may include damage or dysfunction of an afferent and / or efferent nervous system of a subject. A neurological disorder may include damage or dysfunction of a sympathetic and / or parasympathetic nervous system of a subject. A neurological disorder may include damage of dysfunctionPATENT
[0972] ATTORNEY-DOCKET NO.: 51478-036WO4
[0973] of one or more cranial nerves (e.g., the olfactory nerve, optic nerve, oculomotor nerve, trochlear nerve, trigeminal nerve, abducens nerve, facial nerve, vestibulocochlear nerve, glossopharyngeal nerve, vagus nerve, accessory nerve, and / or hypoglossal nerve) of a subject. A neurological disorder may be neuropathic pain which may include inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia. A neurological disorder may be a neurotraumatic disorder, which may include a spinal cord injury, traumatic brain injury, stroke, nerve injury, or peripheral nerve injury. A neurological disorder may be a neurodegenerative disorder (e.g., multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia). A neurological disorder may be epilepsy which may include refractory epilepsy, developmental and epileptic encephalopathies, Lennox-Gastaut syndrome, neurotrauma associated epilepsy (ischemia, stroke, traumatic brain injury), status epilepticus, tumor associated epilepsy, or hypoxic-ischemic encephalopathy. A neurological disorder may be dementia induced psychosis. A neurological disorder may be a neurodevelopmental disorder, which may include an autism spectrum disorder, Rett syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, 22q11.2 microdeletion syndrome, cerebral palsy, Down syndrome, Dravet syndrome, epilepsy (e.g., temporal lobe epilepsy), or sudden unexpected death in epilepsy. A neurological disorder may include an affective disorder, which may include schizophrenia, bipolar-disorder, anxiety disorder, major depressive disorder, attention-deficit / hyperactivity disorder, and the like thereof.
[0974] As used herein, the term “pharmaceutical composition” refers to an active compound, formulated together with one or more pharmaceutically acceptable excipients. In some embodiments, a compound of the disclosure is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, pharmaceutical compositions may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions) or tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, or pastes for application to the tongue.
[0975] The term “pharmaceutically acceptable excipient,” as used herein, refers to any inactive ingredient (for example, a vehicle capable of suspending or dissolving the active compound) having the properties of being nontoxic and non-inflammatory in a subject. Typical excipients include, for example: antiadherents, antioxidants, binders, coatings, compression aids, dis integrants, dyes, emollients, emulsifiers, diluents, film formers or coatings, flavors, fragrances, glidants, lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Excipients include, but are not limited to: butylated optionally substituted hydroxytoluene (e.g., BHT), calcium carbonate, calcium phosphate dibasic, calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxypropyl cellulose, optionally substituted hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starchPATENT
[0976] ATTORNEY-DOCKET NO.: 51478-036WO4
[0977] glycolate, sorbitol, starch, stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those of ordinary skill in the art are familiar with a variety of agents and materials useful as excipients.
[0978] As used herein, the term “pharmaceutically acceptable salt” represents those salts of the compounds described that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P. H. Stahl and C. G. Wermuth), Wiley-VCH, 2008. These salts may be acid addition salts involving inorganic or organic acids. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base group with a suitable acid. Methods for preparation of the appropriate salts are well-established in the art.
[0979] The term “subject,” as used herein, can be a human, non-human primate, or other mammal, such as but not limited to dog, cat, horse, cow, pig, goat, monkey, rat, mouse, and sheep. In preferred embodiments, the subject is a human.
[0980] As used herein, the term “therapeutically effective amount” refers to an amount sufficient to effect beneficial or desired results, such as clinical results, and, as such, a “therapeutically effective amount” depends upon the context in which it is being applied. For example, in the context of administering a compound disclosed herein (e.g., a compounds of any one of Formulas (I), (l-A) to (I- M), or Table 1, and other compounds disclosed herein) to treat a neurological disorder, a therapeutically effective amount of a compound is, for example, an amount sufficient to reverse alleviate the neurological disorder.
[0981] As used herein, the terms “treat” and “treating” refer to a therapeutic treatment of a neurological disorder in a subject. The effect of treatment can include reversing, alleviating, reducing severity of, inhibiting the progression of, reducing the likelihood of recurrence of the neurological disorder or one or more symptoms or manifestations of the neurological disorder, stabilizing (i.e., not worsening) the state of the neurological disorder as compared to the state and / or the condition of the disease or disorder in the absence of the therapeutic treatment.
[0982] Chemical Terms
[0983] The terminology employed herein is for the purpose of describing particular embodiments and is not intended to be limiting.
[0984] For any of the following chemical definitions, a number following an atomic symbol indicates that total number of atoms of that element that are present in a particular chemical moiety. As will be understood, other atoms, such as H atoms, or substituent groups, as described herein, may be present, as necessary, to satisfy the valences of the atoms. For example, an unsubstituted C2 alkyl group has the formula -CH2CH3. When used with the groups defined herein, a reference to the number of carbon atoms includes the divalent carbon in acetal and ketal groups but does not include the carbonyl carbon in acyl, ester, carbonate, or carbamate groups. A reference to the number of oxygen, nitrogen, or sulfur atoms in a heteroaryl group only includes those atoms that form a part of a heterocyclic ring.PATENT
[0985] ATTORNEY-DOCKET NO.: 51478-036WO4
[0986] Those skilled in the art will appreciate that certain compounds described herein can exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, atropisomers, tautomers) or isotopic (e.g., in which one or more atoms have been substituted with a different isotope of the atom, such as hydrogen substituted for deuterium) forms. Unless otherwise indicated or clear from context, a depicted structure can be understood to represent any such isomeric or isotopic form, individually or in combination.
[0987] Compounds described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated.
[0988] Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically active starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separated isomeric forms.
[0989] In some embodiments, one or more compounds depicted herein may exist in different tautomeric forms. As will be clear from context, unless explicitly excluded, references to such compounds encompass all such tautomeric forms. In some embodiments, tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton. In certain embodiments, a tautomeric form may be a prototropic tautomer, which is an isomeric protonation state having the same empirical formula and total charge as a reference form. Examples of moieties with prototropic tautomeric forms are ketone — enol pairs, amide — imidic acid pairs, lactam — lactim pairs, amide — imidic acid pairs, enamine — imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. In some embodiments, tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution. In certain embodiments, tautomeric forms result from acetal interconversion.
[0990] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as2H,3H,11C,13C,14C,13N,15N,150,17O,18O,32P,33P,35S,18F,36CI,123l and125l. Isotopically-labeled compounds (e.g., those labeled with3H and14C) can be useful in compound or substrate tissue distribution assays. Tritiated (i.e.,.sup.3H) and carbon-14 (i.e.,14C)) isotopes can be useful for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e.,2H or D) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced by2H, D, or3H, or one or more carbon atoms are replaced by13C- or14C-enriched carbon. Positron emitting isotopes such as150,13N,11C, and18F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Preparations of isotopically labelled compounds are known to those of skill in the art. For example, isotopically labeled compounds can generally be prepared by following proceduresPATENT
[0991] ATTORNEY-DOCKET NO.: 51478-036WO4
[0992] analogous to those disclosed for compounds of the present invention described herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.
[0993] As is known in the art, many chemical entities can adopt a variety of different solid forms such as, for example, amorphous forms or crystalline forms (e.g., polymorphs, hydrates, solvate). In some embodiments, compounds of the present invention may be utilized in any such form, including in any solid form. In some embodiments, compounds described or depicted herein may be provided or utilized in hydrate or solvate form.
[0994] At various places in the present specification, substituents of compounds of the present disclosure are disclosed in groups or in ranges. It is specifically intended that the present disclosure includes each and every individual subcombination of the members of such groups and ranges. For example, the term “C1-C4 alkyl” is specifically intended to individually disclose methyl, ethyl, C3 alkyl, and C4 alkyl. Furthermore, where a compound includes a plurality of positions at which substituents are disclosed in groups or in ranges, unless otherwise indicated, the present disclosure is intended to cover individual compounds and groups of compounds (e.g., genera and subgenera) containing each and every individual subcombination of members at each position.
[0995] The term “alkyl,” as used herein, refers to a branched or straight-chain monovalent saturated aliphatic radical containing only C and H when unsubstituted. The monovalency of an alkyl group does not include the optional substituents on the alkyl group. For example, if an alkyl group is attached to a compound, monovalency of the alkyl group refers to its attachment to the compound and does not include any additional substituents that may be present on the alkyl group. In some embodiments, the alkyl group may contain, e.g., 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, or 1-2 carbon atoms (e.g., C1-C20, C1-Cis, C1-C16, C1-C14, C1-C12, C1-C10, Ci-Cs, Ci-Ce, C1-C4, or C1-C2). Examples include, but are not limited to, methyl, ethyl, isopropyl, isobutyl, sec-butyl, and tert-butyl. An alkylene is a divalent alkyl group.
[0996] The term “alkenyl,” as used herein, alone or in combination with other groups, refers to a straight chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6 carbon atoms, or 2 carbon atoms).
[0997] The term “aryl,” as used herein, refers to any monocyclic or fused ring bicyclic or multicyclic system containing only carbon atoms in the ring(s), which has the characteristics of aromaticity in terms of electron distribution throughout the ring system, e.g., phenyl, naphthyl, or phenanthryl. An aryl group may have, e.g., six to sixteen carbons or six to fourteen carbons (e.g., six carbons, ten carbons, thirteen carbons, fourteen carbons, or sixteen carbons). An “arylene” is a divalent aryl group.
[0998] The term “arylalkyl,” as used herein, represents an alkyl group substituted with an aryl group. Unsubstituted arylalkyl groups contain from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as Ci-Ce alkyl Ce-C-io aryl, C1-C10 alkyl Ce-C-io aryl, or C1-C20 alkyl Ce-C-io aryl), such as, benzyl and phenethyl. In some embodiments, the alkyl and the aryl are each further substituted with 1, 2, 3, or 4 substituent groups, valency permitting, as defined herein for the respective groups.
[0999] The term “carbocycle,” as used herein, refers to a monovalent, saturated (“cycloalkyl”) or unsaturated, non-aromatic cyclic group containing only C and H when unsubstituted. A carbocycle may have, e.g., three to twenty carbons (e.g., a C3-C7, C3-C8, C3-C9, C3-C10, C3-C11, C3-C12, C3-C14, C3-C16, C3-C18, or C3-C20 carbocycle). Carbocyclyl structures include cycloalkyl groups and unsaturated carbocyclyl radicals.PATENT
[1000] ATTORNEY-DOCKET NO.: 51478-036WO4
[1001] The term “cycloalkyl,” as used herein, refers to a monovalent, saturated cyclic group containing only C and H when unsubstituted. A cycloalkyl group may have, e.g., three to twenty carbons (e.g., a C3-C7, C3-C8, C3-C9, C3-C10, C3-C11, C3-C12, C3-C14, C3-C16, C3-C18, or C3-C20 cycloalkyl). Cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. The term “cycloalkyl” also includes cyclic groups having a bridged multicyclic structure in which one or more carbons bridges two non-adjacent members of a monocyclic ring, e.g., bicyclo[2.2.1]heptyl and adamantyl. The term “cycloalkyl” also includes bicyclic, tricyclic, and tetracyclic fused ring structures, e.g., decalin and spiro-cyclic compounds.
[1002] The term “halo,” as used herein, refers to a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) radical.
[1003] The term “heteroalkyl,” as used herein, refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkyl groups. Examples of heteroalkyl groups are an “alkoxy” which, as used herein, refers alkyl-O- (e.g., methoxy and ethoxy). A heteroalkylene is a divalent heteroalkyl group.
[1004] The term “heteroaryl,” as used herein, refers to an aromatic mono- or polycyclic radical of 5 to 12 atoms having at least one aromatic ring containing 1, 2, or 3 ring atoms selected from nitrogen, oxygen, and sulfur, with the remaining ring atoms being carbon. One or two ring carbon atoms of the heteroaryl group may be replaced with a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl, and thiazolyl. A “heteroarylene” is a divalent heteroaryl group.
[1005] The term “heterocycle,” as used herein, represents a monocyclic or fused ring system, bicyclic or multicyclic, having at least one heteroatom: selected from nitrogen, oxygen, and sulfur, as a ring atom. For example, a heterocycle ring may have, e.g., one to fifteen carbons ring atoms (e.g., a C1-C2, C1-C3, C1-C4, C1-C5, Ci-Ce, C1-C7, Ci-Cs, C1-C9, C1-C10, C1-C11, C1-C12, C1-C13, C1-C14, or C1-C15 heterocycle) and one or more (e.g., one, two, three, four, or five) ring heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Heterocycle groups may or may not include a ring that is aromatic. An aromatic heterocycle group is referred to as a “heteroaryl” group. In preferred embodiments of the disclosure, a heterocycle group is a 3- to 8-membered ring, a 3- to 6-membered ring, a 4- to 6-membered ring, a 4- to 7-membered ring, a 6- to 10-membered ring, a 6- to 12-membered ring, a 5-membered ring, a 6-membered ring, or a 7-membered ring. Exemplary 5-membered heterocycle groups include, for example, pyrrolidine, pyrazolidine, imidazolidine, oxazolidine, isoxazolidine, and thiazolidine. Exemplary 6-membered heterocycle groups include, for example, optionally substituted piperidine, optionally substituted piperazine, morpholine, thiomorpholine, optionally substituted 2 / 7-pyran, optionally substituted 4 / 7-pyran, and optionally substituted tetrahydropyran. Exemplary 7-membered heterocycle groups include optionally substituted azepine, optionally substituted 1,4-diazepine, and optionally substituted 1,4-thiazepine. The term “heterocycle” also includes bicyclic, tricyclic, and tetracyclic fused ring structures containing one or more heteroatoms, e.g., ketals, spirocycles, and bridged compounds.
[1006] The term “optionally substituted X” (e.g., “optionally substituted alkyl”) is intended to be equivalent to “X, wherein X is optionally substituted” (e.g., “alkyl, wherein said alkyl is optionally substituted”). It is notPATENT
[1007] ATTORNEY-DOCKET NO.: 51478-036WO4
[1008] intended to mean that the feature “X” (e.g., alkyl) per se is optional. As described herein, certain compounds of interest may contain one or more “optionally substituted” moieties. In general, the term “substituted”, whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent, e.g., any of the substituents or groups described herein. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. For example, in the term “optionally substituted Ci-Ce alkyl-C2-Cg heteroaryl,” the alkyl portion, the heteroaryl portion, or both, may be optionally substituted. Combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds.
[1009] The term “oxo,” as used herein, represents an =O group.
[1010] The term “stable”, as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow fortheir production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.
[1011] The term “thiol,” as used herein, represents an -SH group.
[1012] Alkyl, carbocycle, cycloalkyl, aryl, heteroalkyl, heteroaryl, and heterocycle groups may be substituted (for example, where the term “optionally substituted” defines additional groups connected to an atom of any of the aforementioned groups) with C3-C12 carbocycle (e.g., C3-C12 cycloalkyl); C5-C12 aryl; heterocycle; halo (halogen); ORa, in which Rais H, C1-C4 alkyl, C1-C4 alkenyl, C1-C4 alkynyl, C3-C12 carbocycle (e.g., cycloalkyl), C5-C12 aryl, or C3-C12 heterocycle; SRb, in which Rbis as defined herein; CN; NO2; N3; NRcRd, in which each of Rcand Rdis, independently, H, C1-C4 alkyl, C1-C4 alkenyl, C1-C4 alkynyl, C3-C12 carbocycle (e.g., cycloalkyl), C5-C12 aryl, or C3-C12 heterocycle; COORa; CONRcRd;
[1013] SO2Re, in which Reis H, C1-C4 alkyl or C5-C12 aryl; SO2NRf'R9', in which each of Rfand R9is, independently, H, C1-C4 alkyl, or C5-C12 aryl; SORh, in which Rhis H, C1-C4 alkyl, or C5-C12 aryl; or P(O)(OR')2, in which each R' is, independently, H or C1-C4 alkyl. Aryl, carbocycle (e.g., cycloalkyl), heteroaryl, and heterocycle groups may also be substituted with C1-C4 alkyl, C1-C4 alkenyl, or C1-C4 alkynyl. Alkyl, alkylene, alkenyl, alkynyl, heteroaryl, carbocycle (e.g., cycloalkyl), and heterocycle groups may also be substituted with oxo or =NR, in which is H or C1-C4 alkyl. In some embodiments, a substituent is further substituted as described herein. For example, a Ci alkyl group, i.e., methyl, may be substituted with oxo to form a formyl group and further substituted with OH or NRCRdto form a carboxyl group or an amido group (e.g. COOH or CONRcRd).
[1014] For the avoidance of doubt, any and all disclosures of methods of treatment or prevention provided herein should also be read as disclosing the compounds, pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same, for use in the described methods of treatment or prevention.
[1015] DETAILED DESCRIPTION OF THE INVENTION
[1016] Described herein are compounds, compositions, and methods for treating neurological disorders, e.g., neurotraumatic disorders, neurodevelopmental disorders, or affective disorders, in a subject. Without wishing to be bound by theory, the compounds described herein may function as KCC2 potentiators. ThePATENT
[1017] ATTORNEY-DOCKET NO.: 51478-036WO4
[1018] compounds described herein are useful for treating neurological disorders, e.g., neurotraumatic disorders, neurodevelopmental disorders, or affective disorders.
[1019] Compounds
[1020] The present disclosure provides compounds and compositions that can be administered to a subject (e.g., a human) in order to treat a neurological disorder (e.g., neurological pain, a neurotraumatic disorder, a neurodevelopmental disorder, or an affective disorder).
[1021] In the first aspect, the disclosure provides a compound having the structure of Formula (I):
[1022]
[1023] Formula (I),
[1024] or a pharmaceutically acceptable salt thereof, wherein:
[1025] R1is optionally substituted C5-C12 aryl, optionally substituted C3-C12 heteroaryl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[1026] R2is H, optionally substituted C1-C4 alkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[1027] R3is H, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[1028] R4is H, optionally substituted C1-C5 alkyl, optionally substituted C1-C5 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;
[1029] R5is optionally substituted C3-C12 heteroaryl, optionally substituted C3-C12 heterocycle, NR6R7, or OR9;
[1030] R6is H or C1-C6alkyl;
[1031] R7is optionally substituted C1-C6 alkyl, C1-C6 heteroalkyl, or optionally substituted C3-C12 heterocycle, wherein the alkyl or heterocycle is optionally substituted with 1 or 2 substituents selected from F, OH, C1-C3 alkoxy, and (CH2)nR8;
[1032] R8is 5- or 6-membered heteroaryl, 3- to 5-membered cycloalkyl, or 3- to 6-membered heterocycle;
[1033] R9is C3-C12 heterocycle; and
[1034] n is 1, 2, or 3.
[1035] In some embodiments, the compound of Formula (I) has the structure:
[1036]
[1037] Formula (l-A), Formula (l-B),PATENT
[1038] ATTORNEY-DOCKET NO.: 51478-036WO4
[1039] Formula (l-M),
[1040]
[1041] O Formula (l-N),0Formula (l-O),
[1042]
[1043] PATENT
[1044] ATTORNEY-DOCKET NO.: 51478-036WO4
[1045]
[1046] Formula (l-AF),PATENT
[1047] ATTORNEY-DOCKET NO.: 51478-036WO4
[1048]
[1049] PATENT
[1050] ATTORNEY-DOCKET NO.: 51478-036WO4
[1051] 0 Formula (ll-AU), o Formula (ll-AV),
[1052]
[1053]
[1054] Formula (ll-BA), Formula (ll-BB),PATENT
[1055] ATTORNEY-DOCKET NO.: 51478-036WO4
[1056]
[1057] PATENT
[1058] ATTORNEY-DOCKET NO.: 51478-036WO4
[1059] Formula (II-BU-1),
[1060]
[1061] Formula (II-BV-1),PATENT
[1062] ATTORNEY-DOCKET NO.: 51478-036WO4
[1063] Formula (II-BW),
[1064]
[1065] Formula (II-BX), Formula (II-BY),
[1066]
[1067] Formula (II-BZ), or a pharmaceutically acceptable salt thereof.
[1068] In some embodiments, the compound is one of the compounds 1-210 of Table 1, or a pharmaceutically acceptable salt thereof.
[1069] Pharmaceutical compositions
[1070] A pharmaceutical composition of the disclosure contains one or more of the compounds disclosed herein (e.g., one or more of the compounds of any one of Formula (I), Formulas (l-A) to (ll-BZ), and other compounds disclosed herein) as the therapeutic compound. In addition to a therapeutically effective amount of the compound, the pharmaceutical compositions also contain a pharmaceutically acceptable excipient, which can be formulated by methods known to those skilled in the art. The compounds disclosed herein (e.g., any one of the compounds of Formula (I), Formulas (l-A) to (ll-BZ), and Table 1, and other compounds disclosed herein) may also be administered with or without other therapeutics for a particular condition, formulated in the same composition or different compositions for administration via the same or different routes.
[1071] The compounds disclosed herein (e.g., any one of the compounds of Formula (I), Formulas (l-A) to (ll-BZ), and Table 1), may be used in the form of free base, or in the form of salts or solvates. All forms are within the scope of the disclosure.
[1072] Routes of administration of the pharmaceutical compositions (or the compounds of the composition) include oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intra-arterial, intracranial, subcutaneous, intracerebroventricular, intraorbital, intraventricular, intrathecal (intraspinal), intraperitoneal, intranasal, inhalation, and topical administration.
[1073] Neurological disorders
[1074] Neurological disorders are disorders that affect the brain, as well as nerves throughout the body and also the spinal cord. Common symptoms of neurological disorders include numbness, tingling, muscle weakness, loss of muscle tone, loss of sensation, disruption or loss of autonomic function, numbness, bowel, or bladder incontinence, paralysis, confusion, pain, altered levels of consciousness,PATENT
[1075] ATTORNEY-DOCKET NO.: 51478-036WO4
[1076] mood disorders, and sexual dysfunction. Certain primary symptoms, such as impaired movement and sensation, can further lead to secondary symptoms including muscle atrophy, loss of voluntary motor control and spasticity at sites of the body innervated by the neurological disorder, pressure (e.g., bed) sores, infections, and respiratory problems. Furthermore, cell death at the neurological disorder may continue long after the initial insult that precipitated the neurological disorder as a result of stress and inflammatory signaling that leads to further ischemia, inflammation, swelling, and disruption of synaptic signaling. Neurological disorder may result in total loss of motor and sensory function distal to the neurological disorder, or incomplete, resulting in partial loss of motor and sensory function.
[1077] Neurological disorders may present as various distinct conditions, depending on the site and severity of the condition. For example, peripheral neurological disorder results from damage to peripheral nerves that extend to the extremities of an individual, leading to numbness and / or loss of sensory function. Proximal neurological disorder results from damage to peripheral and / or central nerves, leading to muscle weakness in the upper part of the legs, buttocks, and / or hips in a subject. Autonomic neurological disorder results from damage and / or dysfunction of autonomic nerves that least to reduced and / or uncontrolled body homeostasis of an individual. Focal neurological disorder and / or polyneurological disorder results from damage to one nerve and / or a plurality of nerves, respectively. Central cord syndrome frequently results from damage to the cervical spinal cord, resulting in weakness in the upper extremities with relative sparing of function in the legs and spared sensation in sacral dermatomes (e.g., urinary sphincter, anal sphincter, and genitalia).
[1078] Neurological disorders include, but are not limited to neurotraumatic disorders such as traumatic brain injury (TBI), stroke, nerve injury or spinal cord injury (SCI). Neurological disorders include, but are not limited to neurodegenerative disorder such as multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia. Neurological disorders include, but are not limited to neurodevelopmental disorders such as autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1, 22q11.2 microdeletion syndrome, or cerebral palsy. Neurological disorders include, but are not limited to neurological pain such as neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia. Neurological disorders include, but are not limited to affective disorders such as schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder. Neurological disorders include, but are not limited to dementia, Lewy body dementia, frontotemporal dementia (FTD), or dementia induced psychosis.
[1079] Dementia refers to general mental deterioration due to organic or psychological causes; characterized by disorientation, impaired memory judgment, and intelligence, and shallow, unstable affect. Dementia includes vascular dementia, ischemic vascular dementia (IVD), frontotemporal dementia (FTD), Lewy body dementia, and Alzheimer's dementia.
[1080] Frontotemporal dementia (FTD) is a neurodegenerative disorder characterized by progressive neurodegeneration in the frontal and temporal lobes of the cerebral cortex which is characterized by a complex clinical presentation that may include deficits in speech comprehension and production, poor motor planning and coordination, and / or loss of executive function characterized by lack of impulsePATENT
[1081] ATTORNEY-DOCKET NO.: 51478-036WO4
[1082] control and a preference for perseverative behaviors. Histological analyses of post-mortem brain tissue from FTD patients exhibit complex and heterogeneous neuropathological profiles with the common presentation of degeneration of neural tissue in the frontal and temporal lobes of the brain. The clinical manifestation of FTD is complex and heterogeneous, but may present as progressive aphasia, decline in cognition (e.g. reduced working memory and executive function), diminished impulse control, emergence of perseverative behaviors, apraxia, apathy, and / or social withdrawal. Neuronal loss in brains of FTD patients is associated with either one of two distinct neuropathologies: 1) the presence of tau-positive neuronal and glial inclusions; or 2) ubiquitin (ub)-positive and TAR DNA-binding protein 43 (TDP43)-positive, but tau-negative inclusions.
[1083] Neurotraumatic disorders or neurotraumatic injury are disorders of the nervous system that result from neurological trauma, such as, e.g., TBI, SCI, PNI, PN, stroke, ischemia, hypoxic-ischemic encephalopathy, tumor-associated epilepsy, and spasticity. In the U. S., roughly 1.7 million people are estimated to suffer TBI every year from causes such as falls, motor vehicle-related incidents, sports injuries, and violence, roughly 52,000 of which succumb to such injuries. Survivors of neurological trauma often face prolonged or indefinite disability.
[1084] TBI (also known as intracranial injury) usually results from an external force suddenly impacting the head of an individual, with the severity of the from mild (e.g., concussion) to severe (e.g., penetrating injury, coma-inducing injury). Sequalae of TBI often includes loss of consciousness, physical, cognitive, social, emotional, and behavioral impairments, but can also be fatal.
[1085] SCI refers to any insult to any region of the spinal cord, e.g., the cervical vertebrae, the thoracic vertebrae, the lumbar vertebrae, the sacral vertebrae, the sacrum, or the coccyx, that causes a negative effect on the function of the spinal cord, e.g., reduce mobility of feeling in limbs. The severity of a spinal cord injury is measured in levels of the injury’s outcome, e.g., ranging from no effect on mobility, e.g., retained walking capacity, to paraplegia (e.g., paralysis of legs and lower region of body), and tetraplegia (e.g., loss of muscle strength in all four extremities).
[1086] PNI refers to any disorder resulting from a nerve injury caused by a traumatic event. Peripheral nerve injury is generally divided into three distinct events, namely, (1) Wallerian degeneration; (2) axon regeneration / growth; and (3) nerve innervation. Types of PNI include, from least severe to most severe: neurapraxia (axon remains intact, but myelin is damaged), axonotmesis (disruption of the axon with maintenance of the epineurium), and neurotmesis (loss of axon continuity / axon transection).
[1087] Stroke is a condition which occurs when the blood supply to a part of the brain is interrupted (i.e., ischemic stroke) by obstruction of a blood vessel by a blood clot, an embolism, systemic hypoperfusion, or cerebral venous sinus thrombosis or when a blood vessel in the brain bursts and releases blood into the spaces surrounding the brain cells (i.e., hemorrhagic stroke) as a result of an intracerebral or a subarachnoid hemorrhage. Stroke poses a substantial public burden as nearly 77.2 million people experienced an ischemic stroke, and 29.1 million people experienced a hemorrhagic stroke in 2019. Depending on the area of the brain affected by the stroke, the symptoms of a stroke may include numbness or weakness, especially on one side of the body corresponding to the contralateral side of the stroke, muscle flaccidity or spasticity, confusion, trouble understanding or producing speech, impaired vision in both eyes, impaired mobility, dizziness, severe headache, or loss of balance or coordination.PATENT
[1088] ATTORNEY-DOCKET NO.: 51478-036WO4
[1089] Neurological trauma may also result from progressive neurodegenerative disorders that results in damage to neural tissues of the CNS. Non-limiting examples of neurodegenerative disorders contemplated for treatment using the presently disclosed compositions and methods include, but are not limited to, amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), Alzheimer's Disease (AD), Huntington’s disease, Lewy body dementia, and frontotemporal dementia.
[1090] Neurodevelopmental disorders refer to neurological disorders resulting from abnormal development of the nervous system and are characterized by abnormal brain function, including, but not limited to, impairments in emotional regulation, learning and memory, impulse control, and cognition. This class of neurological disorders is characterized by diverse etiologies that may account for the multeity of symptoms and their degree of severity. Generally, neurodevelopmental disorders are caused by disruptions of the neurotypical developmental trajectory of the nervous system, which can produce pathological anatomical architecture and connectivity in the nervous system. The causes of neurodevelopmental disorders may include genetic and metabolic diseases, social isolation, inflammatory and autoimmune disorders, infectious diseases, malnutrition, physical trauma, as well as environmental factors. The present disclosure contemplates treatment of neurodevelopmental disorders such as, e.g., autism spectrum disorders, Rett syndrome, Fragile X syndrome, Angelman syndrome, cerebral palsy, Down syndrome, pain (e.g., neuropathic pain, chronic pain, or inflammatory pain), Dravet syndrome, epilepsy (e.g., epilepsy related to one or more KCC2 mutations or epilepsy of infancy with migrating focal seizures (EIMFS) or temporal lobe epilepsy), and sudden unexpected death in epilepsy by administering a composition of the disclosure to the afflicted subject, thereby treating the subject.
[1091] Affective disorders (also known as mood disorders) are a class of neurological conditions characterized by dysregulation of normal affect and mood. Disorders of affect may feature mania or hypomania (e.g., schizophrenia and bipolar disorder), depressed mood (e.g., schizophrenia, bipolar disorder, and MDD), and moods that cycle between mania and depression (e.g., bipolar disorder).
[1092] Affective disorders that may be treated using the disclosed methods and compositions include schizophrenia, bipolar disorder, and MDD.
[1093] Schizophrenia is a psychiatric disease characterized by recurrent psychosis. Symptoms of schizophrenia may include (1) positive symptoms related to hallucinations and reality distortion; (2) disorganized symptoms characterized by attentional impairment and thought disorder; and (3) negative symptoms such as apathy, anhedonia, avolition and loss of verbal fluency. Dysfunction of the limbic-cortical system may be implicated in all three types of symptoms. Causes of schizophrenia have been attributed to biological sex, genetic mutations, environmental factors, malnutrition during pregnancy, and age of parents, among other factors. Several hypotheses exist as to the etiology of schizophrenia, one being the glutamate hypothesis in which reduced glutamatergic drive to potentiatory interneurons is thought to result in reduced cortical inhibition and altered cortical network dynamics that lead to presentation of clinical symptoms.
[1094] Bipolar disorder is an affective disorder that features recurrent bouts of depression and mania (i.e., abnormally elevated mood) spanning from days to weeks each. Causes of bipolar disorder may be manifold, but genetic and environmental factors have been implicated. Generally, two types of bipolar disorder exist, namely, bipolar I disorder, in which there has been at least one manic episode with orPATENT
[1095] ATTORNEY-DOCKET NO.: 51478-036WO4
[1096] without depressive episodes, and bipolar II disorder, in which there has been at least one hypomanic episode and one major depressive episode.
[1097] MDD is a neurological disorder that is often characterized by the patient having at least two weeks of sustained low mood, low self-esteem, loss of interest in routine activities, hyperalgesia, and low psychomotor activity. Depression in MDD may last for periods of time (weeks, days, months, or years) separated by years or may be continuous. MDD may pose a substantial risk to the afflicted patient as the patient may be at a substantially higher risk for suicide. Etiological causes of the disorder have been attributed to substance abuse, other medical conditions (e.g., neurological disorders, metabolic disorders, gastrointestinal disorders, endocrine disorders, cardiovascular disease, pulmonary disease, cancer, and autoimmune disease), and genetic and environmental factors.
[1098] A neurological disorder may also be caused by infection, ischemia, and tumors. Owing to the physiological barriers to regeneration in the central nervous system (CNS), neurological disorders have been a notoriously difficult condition to treat, with most treatments being palliative and rehabilitative. Most treatments involve imposing limitations to movement, maintenance of proper blood pressure by frequent repositioning of the subject, and physical and occupation therapy.
[1099] Methods of treating a neurological disorder
[1100] The compounds disclosed herein (e.g., a compound of Formula (I), Formula (l-A) to (II-E3Z), or Table 1, and other compounds disclosed herein) are, in general, suitable for use in treating a neurological disorder, or complications resulting therefrom.
[1101] The compounds disclosed herein (e.g., a compound of Formula (I), Formula (l-A) to (II-E3Z), or Table 1, and other compounds disclosed herein) are, in general, suitable for use in treating a neurological disorder, e.g., a neurotraumatic, neurodegenerative, neurodevelopmental, neurological pain, epilepsy, and / or affective disorder, or complications resulting therefrom. Non-limiting examples of neurotraumatic disorders may include, but are not limited to traumatic brain injury (TBI), stroke, nerve injury or spinal cord injury (SCI). Non-limiting examples of neurodegenerative disorder may include, but are not limited to multiple sclerosis, amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia. Non-limiting examples of neurodevelopmental disorders may include, but are not limited to autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex (TSC), Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1, 22q11.2 microdeletion syndrome, or cerebral palsy. Non-limiting examples of neurological pain may include, but are not limited to neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, postoperative pain, lower back pain, central pain, fibromyalgia or neuralgia. Non-limiting examples of affective disorders may include, but are not limited to schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder. Neurological disorders include, but are not limited to dementia, Lewy body dementia, frontotemporal dementia (FTD), and dementia induced psychosis.
[1102] The dosage of the pharmaceutical compositions of the disclosure depends on factors including, but are not limited to, the route of administration, the severity of the condition to be treated, and physical characteristics, e.g., age, weight, and general health, of the subject. Typically, the amount of a compoundPATENT
[1103] ATTORNEY-DOCKET NO.: 51478-036WO4
[1104] disclosed herein (e.g., a compound of any one of Formula (I), Formulas (l-A) to (ll-BZ), and other compounds disclosed herein) contained within a single dose may be an amount that effectively imparts the desired therapeutic effect without inducing significant toxicity. The dosage may be adapted by the clinician in accordance with conventional factors such as the extent of the disease and different parameters of the subject.
[1105] The compounds of the disclosure, or pharmaceutical compositions of the disclosure that contain a compound disclosed herein (e.g., a compound of any one of Formula (I), Formulas (l-A) to (ll-BZ), and other compounds disclosed herein), may be administered to a subject in need thereof one time daily or twice daily. Thus, the compounds and pharmaceutical compositions may be administered QD or BID.
[1106] The following examples are merely illustrative and should not be construed as limiting the scope of this disclosure in any way as many variations and equivalents will become apparent to those skilled in the art upon reading the present disclosure. The contents of all references, patents, and patent applications cited throughout this application are expressly incorporated herein by reference.
[1107] EXAMPLES
[1108] The following examples are put forth so as to provide those of ordinary skill in the art with a description of how the compositions and methods described herein may be used, made, and evaluated, and are intended to be purely exemplary of the disclosure and are not intended to limit the scope of what the inventors regard as their disclosure.
[1109] Example 1: (R)-3-isopropyl-8-((4-isopropylphenyl)amino)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 1)
[1110] Step 1. Synthesis of methyl 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylate
[1111] Cl ''"0
[1112] N 5. Z'NH2 CH3ONa, MeOH
[1113] . CK *.. O,.
[1114] Cl N rt, 1 hr Cl N O
[1115]
[1116] O
[1117] To a stirred solution of methyl 5-amino-2,6-dichloropyrimidine-4-carboxylate (1 g, 4.504 mmol, 1 equiv) in anhydrous methanol (10 mL) was added MeONa (0.27 g, 4.954 mmol, 1.1 equiv) at 25 °C. The reaction mixture was stirred at 25°C for a period of 1 h. After completion of reaction, the reaction mixture was quenched by addition of 50 mL of water. The aqueous layer was extracted with ethyl acetate (3 x 100 mL). The combined organic phase was washed with brine (3 x 100 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product, which was further purified by column chromatography using 5% to 55% MeCN in water (0.05% TFA) gradient to afford desired compound methyl 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylate (730 mg, 74.48% yield, 94% purity) as a white solid. MS: MS (ESI) calcd. for C7H8CIN3O3: 217.02, Found: 218.05 [M+H]+PATENT
[1118] ATTORNEY-DOCKET NO.: 51478-036WO4
[1119] Step 2. Synthesis of 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid
[1120] '■'o “'O
[1121] A LiOH, THF / H2O
[1122] .
[1123] cr N if rt, 1 hr
[1124]
[1125] O
[1126] To a solution of methyl 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylate (680 mg, 3.125 mmol, 1 equiv) in THF (6 mL) and H2O (2 mL) was added LiOH (224.52 mg, 9.375 mmol, 3 equiv) at 25°C. The reaction mixture was stirred at 25°C for a period of 1 h. After completion of reaction, the reaction mixture was quenched by addition of 50 mL of water. The aqueous layer was extracted with ethyl acetate (3 x100 mL). The combined organic phase was washed with brine (3 x100 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product, which was further purified by column chromatography using 5% to 55% MeCN in water gradient to afford desired compound 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid (530 mg, 83.31% yield, 92% purity) as a white solid. MS: MS (ESI) calcd. for C6H6ClN3O3: 203.01, Found: 204.10 [M+H]+.
[1127] Step 3. Synthesis of 5-amino-2-chloro-N-isopropyl-6-methoxypyrimidine-4-carboxamide
[1128] H2N^
[1129] HATU, DIEA, DMF
[1130] rt, 1 hr
[1131]
[1132] To a solution of 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid (530 mg, 2.603 mmol, 1 equiv) in DMF (5 mL) were added isopropylamine (307.78 mg, 5.206 mmol, 2 equiv), HATU (1484.86 mg, 3.905 mmol, 1.5 equiv) and DIEA (672.96 mg, 5.206 mmol, 2 equiv) at room temperature. The resulting mixture was stirred at room temperature for 30 mins. The residue was purified by reversed-phase flash chromatography with the following conditions:( column, C18 silica gel; mobile phase, ACN in H2O, 10% to 50% gradient in 10 min; detector, UV 254 nm.) to afford 5-amino-2-chloro-N-isopropyl-6-methoxypyrimidine-4-carboxamide (510 mg, 80.06% yield, 94% purity) as a white solid. MS: MS (ESI) calcd. for C9H13ClN4O2: 244.07, Found: 244.95 [M+H]+
[1133] Step 4. Synthesis of 6-chloro-3-isopropyl-8-methoxypyrimido[5,4-d] [1,3] diazin-4-one
[1134] I A A
[1135] " r
[1136] G..
[1137] HOAc
[1138] 120’C, 16 hrs
[1139]
[1140] To a solution of 5-amino-2-chloro-N-isopropyl-6-methoxypyrimidine-4-carboxamide (490 mg, 2.003 mmol, 1 equiv) in trimethoxymethane (5 mL) was added HOAc (240.52 mg, 4.006 mmol, 2 equiv) at room temperature. The resulting mixture was stirred at 120°C for 16 h. The mixture was allowed to cool down to room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions:( column, C18 silica gel; mobile phase, ACN in H2O, 10% to 50% gradient in 10 min; detector, UV 254 nm.) to afford 6-chloro-3-isopropyl-8-methoxypyrimido[5,4-d] [1,3] diazin-4-one (400 mg,PATENT
[1141] ATTORNEY-DOCKET NO.: 51478-036WO4
[1142] 78.43% yield, 94% purity) as a white solid. MS: MS (ESI) calcd. for C10H11CIN4O2: 254.05, Found: 255.05 [M+H]+
[1143] Step 5. Synthesis of 3-isopropyl-8-methoxy-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one
[1144] 100°C, 7 hrs
[1145]
[1146] A solution of 6-chloro-3-isopropyl-8-methoxypyrimido[5,4-d] [1,3] diazin-4-one (380 mg, 1.492 mmol, 1 equiv), (2R)-2-methylmorpholine (226.39 mg, 2.238 mmol, 1.5 equiv) and DIEA (250.71 mg, 1.940 mmol, 1.3 equiv) in DMSO (4 mL) was stirred at 100°C for 7 hours under nitrogen atmosphere. The mixture was allowed to cool down to room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: (column, C18 silica gel; mobile phase, ACN in H2O, 10% to 80% gradient in 10 min; detector, UV 254 nm.) to afford 3-isopropyl-8-methoxy-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (370 mg, 77.64% yield, 94% purity) as a white solid. MS: MS (ESI) calcd. for C15H21N5O3: 319.16, Found: 320.25 [M+H]+
[1147] Step 6. Synthesis of 8-hydroxy-3-isopropyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one
[1148] OH
[1149] LiCI, TsOH, DMF
[1150] 120'C, 1 hr
[1151]
[1152] To a solution of 3-isopropyl-8-methoxy-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (350 mg, 1.096 mmol, 1 equiv) in DMF (3 mL) were added LiCI (232.28 mg, 5.480 mmol, 5 equiv) and TsOH (943.59 mg, 5.480 mmol, 5 equiv) at room temperature. The resulting mixture was stirred at 120°C for 1h. The mixture was allowed to cool down to room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions:( column, C18 silica gel; mobile phase, ACN in H2O, 10% to 50% gradient in 10 min; detector, UV 254 nm.) to afford 8-hydroxy-3-isopropyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (300 mg, 89.65% yield, 94% purity) as a white solid. MS: MS (ESI) calcd. for C14H19N5O3: 305.14, Found: 306.10 [M+H]+PATENT
[1153] ATTORNEY-DOCKET NO.: 51478-036WO4
[1154] Step 7. Synthesis of 8-chloro-3-isopropyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one
[1155]
[1156] A solution of 8-hydroxy-3-isopropyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (280 mg, 0.917 mmol, 1 equiv) in phosphoryl trichloride (3 mL) was stirred at 80°C for 1 h. The resulting mixture was concentrated under reduced pressure. The reaction was quenched with ice water. The mixture was acidified to pH = 7 with aq. NaHCOs. The resulting mixture was extracted with EA (100 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% NH4HCO3), 5% to 80% gradient in 25 min; detector, UV 254 nm. This resulted in 8-chloro-3-isopropyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (150 mg, 50.52% yield, 93% purity) as a yellow solid. MS: MS (ESI) calcd. for C14H18CIN5O2: 323.11, Found: 324.05 [M+H]+
[1157] Step 8. Synthesis of (R)-3-isopropyl-8-((4-isopropylphenyl)amino)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1158] 100 °C, 16 h
[1159]
[1160] A solution of 8-chloro-3-isopropyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (100 mg, 0.309 mmol, 1 equiv), para-isopropyl aniline (41.76 mg, 0.309 mmol, 1 equiv) and DIEA (51.89 mg, 0.402 mmol, 1.3 equiv) in DMSO (2 mL) was stirred at 100°C for 16h under N2 atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% NH4HCO3), 5% to 80% gradient in 25 min; detector, UV 254 nm. This resulted in (R)-3-isopropyl-8-((4-isopropylphenyl)amino) -6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (87.4 mg, 66.97% yield, 99.4% purity) as a white solid. LC / MS: MS (ESI) calcd. for C23H30N6O2: 422.24. Found: 423.20 [M+H]+ 1H NMR (300 MHz, DMSO-d₆) 6 (ppm): 9.43 - 9.45 (m, 1H), 8.21 - 8.22 (m, 1H), 7.77 - 7.79 (m, 2H), 7.22 - 7.24 (m, 2H), 4.93 - 4.97 (m, 1H), 4.41 - 4.53 (m, 2H), 3.89 - 3.93 (m, 1H), 3.49 - 3.53 (m, 2H), 2.97 - 3.01 (m, 1H), 2.83 - 2.89 (m, 1H), 2.61 - 2.67 (m, 1H), 1.52 - 1.56 (m, 6H), 1.18 - 1.23 (m, 9H).PATENT
[1161] ATTORNEY-DOCKET NO.: 51478-036WO4
[1162] Example 2: (R)-3-ethyl-8-((4-isopropylphenyl)amino)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 3)
[1163] Step 1. Synthesis of 5-amino-2-chloro-N-ethyl-6-methoxypyrimidine-4-carboxamide
[1164] A ivo
[1165] HAW, DIEA, DMF A „NHZ
[1166] A -X OH. *" JI A, N
[1167] cr N rt, 1 hr cr N V >
[1168]
[1169] o 6 '
[1170] To a solution of 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid (1 g, 4.912 mmol, 1 equiv), ethylamine, hydrochloride (600.80 mg, 7.368 mmol, 1.5 equiv) and HATU (3735.48 mg, 9.824 mmol, 2 equiv) in DMF (5 mL) was added DIEA (1904.61 mg, 14.736 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1h under nitrogen atmosphere. The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with water (3 x 40 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (3:7) to afford 5-amino-2-chloro-N-ethyl-6-methoxypyrimidine-4-carboxamide (1.1 g, 97.09% yield, 97% purity) as a white solid. LC / MS: MS (ESI) calcd. for C8H11CIN4O2: 230.06. Found: 231.10 [M+H]+
[1171] Step 2. Synthesis of 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one
[1172] i
[1173] ^Os^. O
[1174] 'o ''O
[1175] „NH2HOACMA.. N„
[1176] 1 1 a. - “ 1 1
[1177] Cl' N' Y > 126'C. 16 hrs Ci 'N Y
[1178]
[1179] O!6:
[1180] To a solution of 5-amino-2-chloro-N-ethyl-6-methoxypyrimidine-4-carboxamide (1 g, 4.336 mmol, 1 equiv) in trimethoxymethane (5 mL) was added HOAc (0.5 mL) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 120 °C for 16h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (900 mg, 86.26% yield, 99% purity) as a white solid. LC / MS: MS (ESI) calcd. for C9H9CIN4O2: 240.04. Found: 241.10 [M+H]+
[1181] Step 3. Synthesis of (R)-3-ethyl-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1182] A'NH
[1183] O, J
[1184] '?1
[1185] N-'Y'A DIEA, DMSO. N.
[1186] A- A N. - *■ ii A
[1187] cr NIf J 100’C, 2 hrs f N N ' y*'
[1188] 0O. > O
[1189]
[1190] I
[1191] To a solution of 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (450 mg, 1.870 mmol, 1 equiv) and (2R)-2-methylmorpholine (226.97 mg, 2.244 mmol, 1.2 equiv) in DMSO (5 mL) wasPATENT
[1192] ATTORNEY-DOCKET NO.: 51478-036WO4
[1193] added DIEA (483.37 mg, 3.740 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100 °C for 2h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-3-ethyl-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (570 mg, 99.83% yield, 91% purity) as a white solid. LC / MS: MS (ESI) calcd. for C14H19N5O3: 305.15. Found: 306.20 [M+H]+
[1194] Step 4. Synthesis of (R)-3-ethyl-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1195] ” O OHwA. x • LICI, TsOH, DM F., XxN,
[1196] x^. A^. AXN, A N r NN1 12O’C, 1 hr f N N y Y
[1197] O1o^J o ’
[1198]
[1199] To a stirred mixture of (R)-3-ethyl-8-methoxy-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (550 mg, 1.801 mmol, 1 equiv) and LiCI (381.78 mg, 9.005 mmol, 5 equiv)) in DMSO (5 mL) was added TsOH (1550.90 mg, 9.005 mmol, 5 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 120 °C for 1h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-3-ethyl-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (500 mg, 95.29% yield, 96% purity) as a white solid. LC / MS: MS (ESI) calcd. for C13H17N5O3: 291.13. Found: 292.20 [M+H]+
[1200] Step 5. Synthesis of (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1201]
[1202] A solution of (R)-3-ethyl-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (500 mg, 1.716 mmol, 1 equiv) in phosphoryl trichloride (1 mL) at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1h under nitrogen atmosphere. The reaction was quenched with water at 0°C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (380 mg, 71.48% yield, 95% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C13H16CIN5O2: 309.10. Found: 310.10 [M+H]+PATENT
[1203] ATTORNEY-DOCKET NO.: 51478-036WO4
[1204] Step 6. Synthesis of (R)-3-ethyl-8-((4-isopropylphenyl)amino)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1205]
[1206] To a solution of (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.323 mmol, 1 equiv) and para-isopropylaniline (52.38 mg, 0.388 mmol, 1.2 equiv) in DMSO (2 mL) was added DIEA (83.45 mg, 0.646 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-3-ethyl-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (62.2 mg, 47.16% yield, 99.7% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C22H28N6O2: 408.23. Found: 409.20 [M+H]+ 1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 9.67 - 9.68 (m, 1H), 8.29 - 8.32 (m, 1H), 7.75 - 7.82 (m, 2H), 7.21 - 7.28 (m, 2H), 4.42 - 4.50 (m, 2H), 4.02 - 4.05 (m, 2H), 3.90 - 3.99 (m, 1H), 3.44 - 3.58 (m, 2H), 2.95 - 3.07 (m, 1H), 2.88 - 2.98 (m, 1H), 2.65 - 2.71 (m, 1H), 1.25 - 1.41 (m, 3H), 1.18 - 1.24 (m, 6H), 1.14 - 1.18 (m, 3H).
[1207] Starting from the same intermediate, (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one, the compounds of Table 2 were synthesized using the corresponding aniline / amine.
[1208] Table 2: Experimental Data for Compounds
[1209] Ex. Cmpd
[1210] Name Structure MS 1H NMR
[1211] # #
[1212] 3 33 (R)-8-((3- 407.15 1H NMR (300 MHz, DMSO- cyclopropylphenyl)am [M+H]+ d6) 6 (ppm): 8.89 - 8.90 (m, ino)-3-ethyl-6-(2- 1H), 8.30 - 8.31 (m, 1H), 7.61 methylmorpholino)pyr - 7.66 (m, 1H), 7.52 - 7.58 (m, imido[5,4-cf]pyrimidin- 1H), 7.21 - 7.27 (m, 1H), 6.87 r-7 NH
[1213] 4(3 / - / )-one - 6.92 (m, 1H), 4.42 - 4.54 (m,
[1214] Ji. • < N
[1215] r?NY > 2H), 3.88 - 4.07 (m, 3H), 3.47 O:
[1216] - 3.52 (m, 2H), 2.97 - 3.11 (m, 1H), 2.70 - 2.76 (m, 1H), 1.86 - 1.94 (m, 1H), 1.16 - 1.30 (m, 6H), 0.92 - 0.99 (m, 2H), 0.64 - 0.72 (m, 2H).
[1217]
[1218] PATENT
[1219] ATTORNEY-DOCKET NO.: 51478-036WO4
[1220] 4 34 (R)-3-ethyl-8-((3-(2- 425.25 1 H NMR (300 MHz, DMSO- hydroxypropan-2- [M+H]+ d6) 6 (ppm): 8.89 - 8.90 (m, yl)phenyl)amino)-6- 1 H), 8.32 - 8.34 (m, 1 H), 8.19 (2- - 8.20 (m, 1 H), 7.50 - 7.53 (m, methylmorpholino)pyr 1 H), 7.29 - 7.34 (m, 1 H), 7.22 imido[5,4-cf]pyrimidin- - 7.24 (m, 1 H), 5.30 - 5.32 (m, 4(3 / - / )-one ' ' AA
[1221] A -A N 1 H), 4.46 - 4.56 (m, 2H), 4.00 A N N A
[1222] 0, > 6 I - 4.07 (m, 2H), 3.88 - 3.92 (m,
[1223] 1 H), 3.47 - 3.51 (m, 2H), 2.99 - 3.07 (m, 1 H), 2.67 - 2.75 (m, 1 H), 1.42 - 1.47 (m, 6H), 1.28 - 1.32 (m, 3H), 1.14 - 1.17 (m, 3H).
[1224] 5 35 (R)-3-ethyl-8-((4- 395.15[ 1 H NMR (400 MHz, DMSO- ethylphenyl)amino)-6- M+H]+ d6) 6: 9.76 (s, 1 H), 8.31 (s, (2- 1 H), 7.73 - 7.80 (m, 2H), 7.20 methylmorpholino)pyr NH - 7.22 (m, 2H), 4.39 - 4.47 (m,
[1225] A N
[1226] imido[5,4-cf]pyrimidin- N A 2H), 3.99 - 4.02 (m, 1 H), 3.51
[1227] . A, N,
[1228] 4(3 / - / )-one rN' if '
[1229] 0. J o
[1230] r!- 3.91 (m, 2H), 3.01 - 3.04 (m,
[1231] 1 H), 2.62 - 2.71 (m, 1 H), 2.57 - 2.60 (m, 2H), 1.29 - 1.36 (m, 3H), 1.15 - 1.29 (m, 6H). 6 36 (R)-3-ethyl-6-(2- 423.25 1 H NMR (300 MHz, MeOD) 6 methylmorpholino)-8- [M+H]+ (ppm): 8.89 - 8.90 (m, 1 H), ((4-(oxetan-3- 8.14 (s, 1 H), 7.85 - 7.87 (m, yl)phenyl)amino)pyri 2H), 7.47 - 7.48 (m, 2H), 5.12
[1232] Ak...
[1233] mido[5,4-cf]pyrimidin- ■ NH - 5.15 (m, 2H), 4.61 - 4.83 (m, 4(3 / - / )-oneWNA - y A 4H), 4.26 - 4.39 (m, 1 H), 4.07 f -x N. A N -A y N |
[1234] < A I J 6 - 4.20 (m, 2H), 3.93 - 4.03 (m,
[1235] 1 H), 3.57 - 3.70 (m, 2H), 2.98 - 3.19 (m, 1 H), 2.74 - 2.76 (m, 1 H), 1.37 - 1.42 (m, 3H), 1.22 - 1.28 (m, 3H)
[1236]
[1237] PATENT
[1238] ATTORNEY-DOCKET NO.: 51478-036WO4
[1239] 7 37 (R)-3-ethyl-8-((3- 397.10 1 H NMR (400 MHz, DMSO- methoxyphenyl)amin [M+H]+ d6) 6: 9.52 (s, 1 H), 8.26 - o)-6-(2- 8.28 (m, 1 H), 7.68 - 7.70 (m, methylmorpholino)pyr 1 H), 7.41 - 7.48 (m, 1 H), 7.23 imido[5,4-cf]pyrimidin- - 7.25 (m, 1 H), 6.65 - 6.67 (m, 4(3 / - / )-one AA
[1240] Ji. -A N 1 H), 4.44 - 4.54 (m, 2H), 4.02 r i
[1241] O. > 0 ’ - 4.03 (m, 2H), 3.92 - 3.99 (m, i 1 H), 3.77 - 3.87 (m, 3H), 3.51
[1242] - 3.77 (m, 2H), 2.92 - 3.03 (m, 1 H), 2.57 - 2.60 (m, 1 H), 1.16 - 1.29 (m, 6H).
[1243] 8 38 (R)-8-((3- 401.10 1 H NMR (400 MHz, DMSO- chlorophenyl)amino)- [M+H]+ d6) 6: 9.88 (s, 1 H), 8.28 - 3-ethyl-6-(2- 8.30 (m, 1 H), 8.20 - 8.22 (m, methylmorpholino)pyr 1 H), 7.77 - 7.83 (m, 1 H), 7.38
[1244] £1
[1245] imido[5,4-cf]pyrimidin- cr NH - 7.40 (m, 1 H), 7.14 - 7.20 (m, 4(3 / - / )-one 1 H), 4.44 - 4.45 (m, 2H), 4.01
[1246] A 1
[1247] r N * r
[1248] O J 0!- 4.19 (m, 2H), 3.92 - 3.99 I (m, 1 H), 3.51 - 3.53 (m, 2H),
[1249] 3.01 - 3.04 (m, 1 H), 2.62 - 2.71 (m, 1 H), 1.29 - 1.36 (m, 3H), 1.15 - 1.19 (m, 3H). 9 39 (R)-8-((4- 407.25 1 H NMR (300 MHz, MeOD) 6 cyclopropylphenyl)am [M+H]+ (ppm): 8.89 - 8.91 (m, 1 H), ino)-3-ethyl-6-(2- 8.36 (s, 1 H), 7.51 - 7.71 (m, methylmorpholino)pyr 2H), 7.12 - 7.22 (m, 2H), 4.48 imido[5,4-cf]pyrimidin- - 4.51 (m, 2H), 4.1 1 - 4.24 (m,
[1250] Xl
[1251] 4(3 / - / )-one NH 2H), 4.02 - 4.04 (m, 1 H), 3.59
[1252] ¥ A N - 3.74 (m, 2H), 3.16 - 3.32 (m, f?NY 1
[1253] O J 0!1 H), 2.91 - 2.95 (m, 1 H), 1.89
[1254] - 2.04 (m, 1 H), 1.44 - 1.46 (m, 3H), 1.22 - 1.24 (m, 3H), 0.93 - 1.08 (m, 2H), 0.66 - 0.78 (m, 2H)
[1255]
[1256] PATENT
[1257] ATTORNEY-DOCKET NO.: 51478-036WO4
[1258] 10 10 (R)-8-((4- 401.15 1 H NMR (300 MHz, MeOD) 6 chlorophenyl)amino)- [M+H]+ (ppm): 8.89 - 8.91 (m, 1 H), 3-ethyl-6-(2- 8.32 (s, 1 H), 7.63 - 7.90 (m,
[1259] CL
[1260] methylmorpholino)pyr ¥,1. 2H), 7.32 - 7.56 (m, 2H), 4.49
[1261] NH
[1262] imido[5,4-cf]pyrimidin- N X -r, N -.>5 - 4.54 (m, 2H), 4.08 - 4.23 (m, 4(3 / - / )-one Y N
[1263] < N N y > 2H), 4.00 - 4.01 (m, 1 H), 3.52 6. o ' - 3.75 (m, 2H), 3.15 - 3.27 (m,
[1264] 1 H), 2.81 - 2.89 (m, 1 H), 1.39 - 1.44 (m, 3H), 1.22 - 1.24 (m, 3H).
[1265] 11 11 (R)-8-((4- 433.20 1 H NMR (300 MHz, DMSO- (difluoromethoxy)phe X
[1266] Q O— s [M+H]+ d6) 6 (ppm): 8.89 - 8.90 (m, nyl)amino)-3-ethyl-6-.0. 1 H), 8.30 (s, 1 H), 7.88 - 7.91 (2- i n ■s-,-. (m, 2H), 7.19 - 7.43 (m, 3H),
[1267] ¥< O~-- methylmorpholino)pyr A N 4.37 - 4.49 (m, 2H), 3.89 - imido[5,4-cf]pyrimidin- r *
[1268] a J a 4.06 (m, 3H), 3.46 - 3.50 (m, f
[1269] 4(3 / - / )-one 2H), 2.96 - 3.06 (m, 1 H), 2.68
[1270] - 2.73 (m, 1 H), 1.21 - 1.29 (m, 3H), 1.12 - 1.20 (m, 3H). 12 12 (R)-8-((4- 417.15[ 1 H NMR (400 MHz, DMSO- (difluoromethyl)pheny M+H]+ d6) 6 (ppm): 9.79 - 9.80 (m, l)amino)-3-ethyl-6-(2- 1 H), 8.26 - 8.27 (m, 1 H), 8.03 methylmorpholino)pyr F - 8.05 (m, 2H), 7.55 - 7.58 (m, imido[5,4-cf]pyrimidin- 2H), 6.85 - 7.13 (m, 1 H), 4.41 4(3 / - / )-one N: ' -W- - 4.48 (m, 2H), 4.02 - 4.04 (m, f N N' Y > 2H), 3.91 - 3.98 (m, 1 H), 3.47 6^J d >
[1271] - 3.52 (m, 2H), 2.96 - 3.03 (m, 1 H), 2.64 - 2.70 (m, 1 H), 1.32 - 1.33(m, 3H), 1.26 - 1.27 (m, 3H).
[1272] 13 13 (R)-3-ethyl-8-((4- 397.20 1 H NMR (400 MHz, DMSO- methoxyphenyl)amin [M+H]+ d6) 6 (ppm): 8.89 - 8.90 (m, o)-6-(2- 1 H), 8.34 - 8.35 (m, 1 H), 7.70 methylmorpholino)pyr - 7.73 (m, 2H), 6.96 - 7.00 (m, imido[5,4-cf]pyrimidin- 2H), 4.34 - 4.46 (m, 2H), 3.93 4(3 / - / )-one - 4.05 (m, 2H), 3.88 - 3.90 (m,
[1273] 1 H), 3.75 - 3.77 (m, 3H), 3.50 - 3.54 (m, 2H), 2.95 - 3.06 (m, 1 H), 2.70 - 2.78 (m, 1 H), 1.28 - 1.33 (m, 3H), 1.14 - 1.17 (m, 3H).
[1274]
[1275] PATENT
[1276] ATTORNEY-DOCKET NO.: 51478-036WO4
[1277] 14 14 (R)-3-ethyl-8-((4-(4- 477.20[ 1 H NMR (300 MHz, DMSO- fluorophenoxy)phenyl M+H] + d6) 6: 9.82 (s, 1 H), 8.30 (s, )amino)-6-(2- 1 H), 7.80 - 7.98 (m, 2H), 7.13 methylmorpholino)pyr. O. - 7.51 (m, 2H), 6.94 - 7.11 (m, imido[5,4-cf]pyrimidin- 4H), 4.37 - 4.49 (m, 2H), 3.97 4(3 / - / )-one... N r N r T’ti > - 4.10 (m, 2H), 3.79 - 3.95 (m,
[1278] 6 J a >
[1279] i 1 H), 3.40 - 3.65 (m, 2H), 2.95
[1280] - 3.02 (m, 1 H), 2.62 - 2.70 (m, 1 H), 1.26 - 1.31 (m, 3H), 1.14 - 1.16 (m, 3H).
[1281] 15 8 (R)-3-ethyl-8-((1- 413.25 1 H NMR (300 MHz, DMSO- isopropyl-1 / 7-pyrazol- [M+H] + d6) 6 (ppm): 8.41 (s, 1 H), 4-yl)amino)-6-(2- 8.14 (s, 1 H), 7.86 (s, 1 H), methylmorpholino)pyr \ 7.60 - 7.65 (m, 1 H),4.94 - / :NH
[1282] imido[5,4-cf]pyrimidin- N 4.96 (m, 1 H), 4.34 - 4.60 (m, 4(3 / - / )-onex-.„ N
[1283] j' N N Y 1 3H), 3.93 - 4.03 (m, 1 H), 3.62 0. > O '
[1284] - 3.82 (m, 2H), 3.21 - 3.25 (m, 1 H), 2.90 - 2.94 (m, 1 H), 1.46 - 1.49 (m, 12H), 1.18 - 1.20 (m, 3H).
[1285] 16 15 (R)-3-ethyl-8-((4- 423.20[ 1 H NMR (300 MHz, DMSO- isopropylphenyl)(met M+H] + d6) 6: 7.80 - 7.82 (m, 1 H), hyl)amino)-6-(2- 7.20 - 7.22 (m, 2H), 7.10 - methylmorpholino)pyr T £ 7.13 (m, 2H), 4.40 - 4.44 (m, imido[5,4-cf]pyrimidin- rvs 2H), 3.83 - 3.88 (m, 3H), 3.56
[1286] X N;
[1287] 4(3 / - / )-one r *wr "i - 3.60 (m, 3H), 3.45 - 3.52 (m,
[1288] I 2H), 2.86 - 3.03 (m, 2H), 2.60
[1289] - 2.68 (m, 1 H), 1.24 - 1.26 (m, 9H), 1.21 - 1.23 (m, 3H).
[1290]
[1291] PATENT
[1292] ATTORNEY-DOCKET NO.: 51478-036WO4
[1293] 17 9 (R)-3-ethyl-6-(2- 464.15 1 H NMR (400 MHz, DMSO- methylmorpholino)-8- [M+H]+d6) 6 (ppm): 8.64 - 8.65 (m, ((1 -phenylpiperidin-4- 1 H), 8.33 - 8.34 (m, 1 H), 7.31 yl)amino)pyrimido[5,4 - 7.33 (m, 2H), 7.29 - 7.30 (m, -cf]pyrimidin-4(3 / 7)- 2H), 6.96 - 7.00 (m, 1 H), 4.91
[1294] Cl
[1295] one L i - 4.95 (m, 1 H), 4.44 - 4.51 (m,
[1296] NH
[1297] 2H), 4.26 - 4.27 (m, 1 H), 3.90.-H A N
[1298] r « N YA - 3.92 (m, 1 H), 3.73 - 3.76 (m, i 2H), 3.48 - 3.55 (m, 2H), 3.02
[1299] - 3.08 (m, 3H), 2.72 - 2.78 (m, 1 H), 1.96 - 2.00 (m, 4H), 1.45 - 1.50 (m, 6H), 1.21 - 1.30 (m, 3H).
[1300] 18 16 (R)-8-((4-cyclopropyl- 425.15 1 H NMR (300 MHz, DMSO- 2- [M+H]+d6) 6 (ppm): 9.11 - 9.15 (m, fluorophenyl)amino)- 1 H), 8.26 - 8.27 (m, 1 H), 7.80 3-ethyl-6-(2- F - 7.85 (m, 1 H), 6.96 - 7.04 (m, methylmorpholino)pyr r c
[1301] NH 2H), 4.31 - 4.45 (m, 2H), 3.85 imido[5,4-cf]pyrimidin- - 4.03 (m, 3H), 3.34 - 3.47 (m, 4(3 / - / )-one N ": 'N■
[1302] oxJ 6 ’ 2H), 2.88 - 2.96 (m, 1 H), 2.50 I - 2.63 (m, 1 H), 1.90 - 1.99 (m,
[1303] 1 H), 1.30 - 1.34 (m, 3H), 1.23 - 1.28 (m, 3H), 0.93 - 0.97 (m, 2H), 0.69 - 0.72 (m, 2H). 19 17 (R)-8- 345.20 1 H NMR (300 MHz, DMSO- (cyclobutylamino)-3- [M+H] + d6) 6 (ppm): 8.02 - 8.29 (m, ethyl-6-(2- 1 H), 7.76 - 8.02 (m, 1 H), 4.32 methylmorpholino)pyr NH - 4.72 (m, 3H), 3.73 - 4.13 (m, imido[5,4-cf]pyrimidin-N1A 3H), 3.42 - 3.58 (m, 2H), 2.78 4(3 / - / )-oneO-A ° - 3.19 (m, 1 H), 2.54 - 2.78 (m,
[1304] 1 H), 2.01 - 2.38 (m, 4H), 1.51 - 1.87 (m, 2H), 1.21 - 1.48 (m, 3H), 0.99 - 1.19 (m, 3H).
[1305]
[1306] PATENT
[1307] ATTORNEY-DOCKET NO.: 51478-036WO4
[1308] 20 18 (R)-8- 359.25 1 H NMR (300 MHz, DMSO- (cyclopentylamino)-3- [M+H] + d6) 6 (ppm): 8.01 - 8.31 (m, ethyl-6-(2- 1 H), 7.31 - 7.68 (m, 1 H), 4.43 methylmorpholino)pyr - 4.62 (m, 2H), 4.29 - 4.61 (m, / ~Y
[1309] imido[5,4-cf]pyrimidin- 1 H), 3.94 - 4.08 (m, 2H), 3.82 4(3 / - / )-one A -Y Y - 3.94 (m, 1 H), 3.42 - 3.55 (m, r?Nr ]
[1310] o, o!2H), 2.81 - 2.99 (m, 1 H), 2.51
[1311] - 2.62 (m, 1 H), 1.89 - 2.09 (m, 2H), 1.49 - 1.89 (m, 6H), 1.22 - 1.29 (m, 3H), 1.1 1 - 1.21 (m, 3H).
[1312] O O Ti—. 21 19 (R)-3-ethyl-6-(2- 435.10 1 H NMR (300 MHz, DMSO-? ’""'‘X / /
[1313] methylmorpholino)-8- [M+H] + d6) 6 (ppm): 9.72 - 9.75 (m,
[1314] Z z
[1315] 1 H), 8.22 - 8.25 (m, 1 H), 8.12 ((4-x
[1316] (trifluoromethyl)pheny - 8.15 (m, 2H), 7.71 - 7.75 (m, J _
[1317] l)amino)pyrimido[5,4- AA 2H), 4.38 - 4.61 (m, 2H), 3.88
[1318] A N
[1319] r *NY 'i cf]pyrimidin-4(3 / - / )-one - 4.13 (m, 3H), 3.47 - 3.53 (m, O. J O
[1320] I 2H), 3.01 - 3.06 (m, 1 H), 2.69
[1321] - 2.73 (m, 1 H), 1.29 - 1.31 (m, 3H), 1.17 - 1.19 (m, 3H). 22 20 (R)-3-ethyl-8-((3- 453.10 1 H NMR (300 MHz, DMSO- fluoro-4- [M+H] + d6) 6 (ppm): 9.70 - 9.75 (m, (trifluoromethyl)pheny 1 H), 8.25 - 8.28 (m, 1 H), 8.11 l)amino)-6-(2- - 8.15 (m, 1 H), 7.98 - 8.01 (m, methylmorpholino)pyr 1 H), 7.74 - 7.79 (m, 1 H), 4.39 imido[5,4-cf]pyrimidin- - 4.50 (m, 2H), 3.92 - 4.00 (m, 4(3 / - / )-one 3H), 3.52 - 3.55 (m, 2H), 3.04
[1322] - 3.09 (m, 1 H), 2.71 - 2.75 (m, 1 H), 1.23 - 1.32 (m, 3H), 1.09 - 1.21 (m, 3H).
[1323] 23 21 (R)-3-ethyl-6-(2- 449.10 1 H NMR (300 MHz, DMSO- methylmorpholino)-8- [M+H] + d6) 6 (ppm):9.68 - 9.75 (m, ((4-(2,2,2- 1 H), 8.20 - 8.32 (m, 1 H), 7.81 trifluoroethyl)phenyl)a - 7.93 (m, 2H), 7.29 - 7.41 (m,
[1324] ”MH
[1325] mino)pyrimido[5,4- 2H), 4.31 - 4.80 (m, 2H), 3.85 cf]pyrimidin-4(3 / - / )-one - 3.47 (m, 3H), 3.62 - 3.70 YY A
[1326] 0. J o '
[1327] (m, 2H), 3.39 - 3.51 (m, 2H), I
[1328] 2.91 - 3.10 (m, 1 H), 2.65 - 2.80 (m, 1 H), 1.27 - 1.38 (m, 3H), 1.01 - 1.23 (m, 3H).
[1329]
[1330] PATENT
[1331] ATTORNEY-DOCKET NO.: 51478-036WO4
[1332] In a similar approach to the one used for Example 2, starting from (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one, (R)-8-chloro-3-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one, 8-chloro-3-ethyl-6-morpholinopyrimido[5,4-d] pyrimidin-4(3H)-one or (R)-8-chloro-3-cyclopropyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one, the compounds of Table 2A were synthesized using the corresponding aniline. Alternate conditions (Cs₂CO₃ in DMF at 80 °C or K2CO3 in DMSO at 100 °C) were used when the previously described conditions failed.
[1333] Table 2A: Experimental Data for Compounds
[1334] Ex Cmpd
[1335] Name Structure MS 1H NMR
[1336] # #
[1337] 24 40 (R)-3-ethyl-8-((4-(2- 425.25 1H NMR (400 MHz, DMSO- hydroxypropan-2- [M+H]+ d6) 6 (ppm): 9.51 - 9.52 (m, yl)phenyl)amino)-6-(2- 1H), 8.25 - 8.26 (m, 1H), 7.79 methylmorpholino)pyri - 7.82 (m, 2H), 7.42 - 7.45 (m, mido[5,4-d]pyrimidin- 2H), 4.97 - 4.98 (m, 1H), 4.50 4(3H)-one N- k - 4.54 (m, 2H), 3.99 - 4.03 (m,
[1338] 6 ^ J O 2H), 3.90 - 3.97 (m, 1H), 3.46 - 3.50 (m, 2H), 2.98 - 2.99 (m, 1H), 2.62 - 2.69 (m, 1H), 1.42 - 1.43 (m, 6H), 1.29 - 1.35 (m, 3H), 1.16 - 1.23(m, 3H).
[1339] 25 41 (R)-8-((4-cyclopropyl-3- 425.25 1H NMR (300 MHz, DMSO- fluorophenyl)amino)-3- [M+H]+ d6) 6 (ppm): 9.65 - 9.66 (m, ethyl-6-(2- 1H), 8.24 - 8.25 (m, 1H), 7.76 methylmorpholino)pyri - 7.84 (m, 1H), 7.61 - 7.65 (m, mido[5,4-d]pyrimidin- A f 1H), 6.94 - 7.00 (m, 1H), 4.39 4(3H)-one - 4.51 (m, 2H), 3.90 - 4.04 (m,
[1340] 3H), 3.46 - 3.53 (m, 2H), 2.99 V6 1- 3.04 (m, 1H), 2.63 - 2.71 (m,
[1341] 1H), 1.96 - 2.02 (m, 1H), 1.25 - 1.30 (m, 3H), 1.15 - 1.23 (m, 3H), 0.90 - 0.97 (m, 2H), 0.69 - 0.73 (m, 2H).
[1342]
[1343] PATENT
[1344] ATTORNEY-DOCKET NO.: 51478-036WO4
[1345] 26 42 (R)-3-ethyl-8-((2-fluoro- 453.10 1 H NMR (400 MHz, DMSO- 4- [M+H]+ d6) 6 (ppm): 9.38 - 9.42 (m, (trifluoromethyl)phenyl) 1 H), 8.31 - 8.36 (m, 2H), 7.80 amino)-6-(2- - 7.83 (m, 1 H), 7.67 - 7.70 (m, methylmorpholino)pyri 1 H), 4.28 - 4.54 (m, 2H), 3.95 mido[5,4-d]pyrimidin- - 4.05 (m, 2H), 3.88 - 3.95 (m, 4(3H)-one 1 H), 3.41 - 3.59 (m, 2H), 2.88
[1346] - 3.03 (m, 1 H), 2.60 - 2.71 (m, 1 H), 1.21 - 1.34 (m, 3H), 1.15 - 1.18 (m, 3H).
[1347] 27 43 (R)-3-ethyl-8-((4-(1- 425.15 1 H NMR (300 MHz, DMSO- fluorocyclopropyl)phen [M+H]+ d6) 6 (ppm): 9.10 - 9.12 (m, yl)amino)-6-(2- 1 H), 7.86 - 7.89 (m, 2H), 7.31
[1348] ZVF
[1349] methylmorpholino)pyri? Z z~ - 7.35 (m, 2H), 4.39 - 4.51 (m, mido[5,4-d]pyrimidin- AY 2H), 3.80 - 4.04 (m, 2H), 3.75 4(3H)-one - 3.78 (m, 1 H), 3.01 - 3.06 (m,
[1350] Y “
[1351] 2H), 2.72 - 2.77 (m, 1 H), 2.72 - 2.77 (m, 1 H), 1.12 - 1.52 (m, 10H).
[1352] 28 44 (R)-8-(benzo[d]thiazol- 424.05 1 H NMR (300 MHz, DMSO- 5-ylamino)-3-ethyl-6- [M+H]+ d6) 6 (ppm): 9.83 - 9.91 (m, (2- 1 H), 9.38 - 9.41 (m, 1 H), 8.68 methylmorpholino)pyri S-VA>. - 8.78 (m, 1 H), 8.23 - 8.25 (m, mido[5,4-d]pyrimidin- 1 H), 8.07 - 8.15 (m, 1 H), 7.92
[1353] J. N
[1354] 4(3H)-one - 8.00 (m, 1 H), 4.32 - 4.65 (m,
[1355] J Q!2H), 3.87 - 4.12 (m, 3H), 3.42 i
[1356] - 3.61 (m, 2H), 2.88 - 3.12 (m, 1 H), 2.62 - 2.75 (m, 1 H), 1.30 - 1.41 (m, 3H), 1.01 - 1.20 (m, 3H).
[1357] 29 45 (R)-8-(benzo[d]thiazol- 424.05 1 H NMR (300 MHz, DMSO- 6-ylamino)-3-ethyl-6- [M+H]+ d6) 6 (ppm): 9.27 - 9.30 (m, (2- 1 H), 8.67 - 8.73 (m, 1 H), 8.25 methylmorpholino)pyri N-A'X-- - 8.27 (m, 1 H), 8.03 - 8.11 (m, mido[5,4-d]pyrimidin- 1 H), 7.92 - 8.00 (m, 1 H), 4.35 4(3H)-one - 4.68 (m, 2H), 3.99 - 4.11 (m,
[1358] I 3H), 3.59 - 3.61 (m, 2H), 2.88
[1359] - 3.08 (m, 1 H), 2.65 - 2.71 (m, 1 H), 1.28 - 1.37 (m, 3H), 1.15 - 1.21 (m, 3H).
[1360]
[1361] PATENT
[1362] ATTORNEY-DOCKET NO.: 51478-036WO4
[1363] 30 46 (R)-8-((3,5- 403.10 1 H NMR (400 MHz, DMSO- difluorophenyl)amino)- [M+H]+ d6) 6 (ppm): 9.98 - 10.07 (m, 3-ethyl-6-(2- F 1 H), 8.30 (s, 1 H), 7.72 - 7.82 methylmorpholino)pyri (m, 2H), 6.95 - 6.98 (m, 1 H), mido[5,4-d]pyrimidin- F 4.40 - 4.51 (m, 2H), 4.00 - K--U. K
[1364] 4(3H)-one 4.05 (m, 2H), 3.91 - 3.99 (m, d > 6 1 H), 3.48 - 3.61 (m, 2H), 2.98
[1365] - 3.12 (m, 1 H), 2.63 - 2.78 (m, 1 H), 1.21 - 1.32 (m, 3H), 1.05 - 1.20 (m, 3H).
[1366] 31 47 (R)-3-ethyl-8-((3-fluoro- 415.15 1 H NMR (400 MHz, DMSO- 5- [M+H]+ d6) 6 (ppm): 9.73 - 9.75 (m, methoxyphenyl)amino) 1 H), 8.30 (s, 1 H), 7.50 - 7.53 -6-(2- F (m, 1 H), 7.41 - 7.48 (m, 1 H), methylmorpholino)pyri 6.49 - 6.55 (m, 1 H), 4.39 - mido[5,4-d]pyrimidin- ft Y 4.55 (m, 2H), 4.00 - 4.08 (m, 3 ) o f M' 4 N Ysi
[1367] 4( H - ne? 2H), 3.92 - 3.95 (m, 1 H), 3.81
[1368] CK J 5
[1369] i (s, 3H), 3.49 - 3.51 (m, 2H),
[1370] 2.98 - 3.08 (m, 1 H), 2.67 - 2.78 (m, 1 H), 1.21 - 1.32 (m, 3H), 1.12 - 1.20 (m, 3H). 32 48 (R)-3-ethyl-8-((4- 411.10 1 H NMR (300 MHz, DMSO- (methoxymethyl)phenyl [M+H]+ d6) 6 (ppm): 9.58 - 9.60 (m, )amino)-6-(2- 1 H), 8.25 - 8.27 (m, 1 H), 7.84 methylmorpholino)pyri - 7.87 (m, 2H), 7.29 - 7.31 (m, mido[5,4-d]pyrimidin- 2H), 4.37 - 4.51 (m, 4H), 4.01 4(3H)-one - 4.03 (m, 2H), 3.92 - 3.99 (m,
[1371] •'■ ■'NA
[1372] 6. J 6 s 1 H), 3.51 - 3.53 (m, 2H), 3.34 i
[1373] - 3.36 (m, 3H), 2.97 - 3.01 (m, 1 H), 2.62 - 2.68 (m, 1 H), 1.32 - 1.35 (m, 3H), 1.28 - 1.30 (m, 3H).
[1374]
[1375] PATENT
[1376] ATTORNEY-DOCKET NO.: 51478-036WO4
[1377] 33 49 (R)-3-((7-ethyl-2-(2- 438.20 1 H NMR (400 MHz, DMSO) 6 methylmorpholino)-8- [M+H]+ 9.89 (s, 1 H), 8.30 (s, 1 H), oxo-7, 8- 8.10 (s, 1 H), 7.89 - 7.83 (m, dihydropyrimido[5,4- 1 H), 7.46 - 7.38 (m, 1 H), 7.15 d]pyrimidin-4- - 7.08 (m, 1 H), 4.51 - 4.39 (m, yl)amino)-N, N- w A N,-i V s
[1378] C ‘ 2H), 4.07 - 3.97 (m, 2H), 3.94 dimethylbenzamide i - 3.86 (m, 1 H), 3.58 - 3.44 (m,
[1379] 2H), 3.05 - 2.92 (m, 7H), 2.72 - 2.62 (m, 1 H), 1.33 - 1.25 (m, 3H), 1.17 - 1.20 (m, 3H). 34 50 (R)-8-((4-(1,1- 431.15 1 H NMR (300 MHz, DMSO- difluoroethyl)phenyl)am [M+H]+ d6) 6 (ppm): 9.80 - 10.00 (m, ino)-3-ethyl-6-(2- 1 H), 8.28 - 8.30 (m, 1 H), 7.96 methylmorpholino)pyri - 8.30 (m, 2H), 7.58 - 7.96 (m, mido[5,4-d]pyrimidin- 2H), 4.39 - 4.51 (m, 2H), 3.90 4(3H)-one - 4.39 (m, 3H), 3.46 - 3.56 (m,
[1380] 2H), 2.98 - 3.06 (m, 1 H), 2.65 - 2.73 (m, 1 H), 1.92 - 2.07 (m, 3H), 1.29 - 1.31 (m, 3H), 1.16 - 1.26 (m, 3H).
[1381] 35 51 (R)-3-ethyl-6-(2- 445.05 1 H NMR (400 MHz, DMSO- methylmorpholino)-8- [M+H]+ d6) 6 (ppm):10.05 (s, 1 H), ((4-(methylsulfonyl)- 8.29 (s, 1 H), 8.17 - 8.23 (m, phenyl)amino)pyrimido[ 2H), 7.88 - 7.94 (m, 2H), 4.38 5,4-d]pyrimidin-4(3H)- - 4.56 (m, 2H), 3.89 - 4.07 (m, one 1 1 3H), 3.45 - 3.60 (m, 2H), 3.20
[1382] (s, 3H), 2.94 - 3.10 (m, 1 H), 2.63 - 2.76 (m, 1 H), 1.29 (t, J =9.4, 3H), 1.19 (d, J =8.2, 3H).
[1383] 36 52 (R)-3-ethyl-6-(2- 445.10 1 H NMR (400 MHz, DMSO- methylmorpholino)-8- [M+H]+ d6) 6 (ppm):10.04 (s, 1 H), ((3-(methylsulfonyl)- 8.96 (s, 1 H), 8.28 (s, 1 H), phenyl)amino)pyrimido[ 7.97 - 8.08 (m, 1 H), 7.58 - 5,4-d]pyrimidin-4(3H)- 7.63 (m, 2H), 4.49 - 4.56 (m, one 2H), 4.01 - 4.10 (m, 2H), 3.86
[1384] - 3.95 (m, 1 H), 3.44 - 3.59 (m, 2H), 3.21 (s, 3H), 2.92 - 3.03 (m, 1 H), 2.62 - 2.74 (m, 1 H), 1.29 (t, J =9.6, 3H), 1.19 (d, J =8.2, 3H).
[1385]
[1386] PATENT
[1387] ATTORNEY-DOCKET NO.: 51478-036WO4
[1388] 37 53 (R)-4-((7-ethyl-2-(2- 438.15 1 H NMR (400 MHz, DMSO- methylmorpholino)-8- [M+H]+ d6) 6 (ppm): 9.86 - 9.87 (m, oxo-7, 8-dihydro- 1 H), 8.29 - 8.30 (m, 1 H), 7.94 q
[1389] pyrimido[5,4- - 7.96 (m, 2H), 7.42 - 7.44 (m,
[1390] I L
[1391] d]pyrimidin-4- '’NH 2H), 4.40 - 4.51 (m, 2H), 4.01 yl)amino)-N, N- - 4.11 (m, 2H), 3.93 - 3.99 (m, dimethylbenzamide 6. J 6 *
[1392] I 1 H), 3.47 - 3.55 (m, 2H), 2.97
[1393] - 3.04 (m, 7H), 2.65 - 2.71 (m, 1 H), 1.27 - 1.30 (m, 3H), 1.16 - 1.17 (m, 3H).
[1394] 38 54 (R)-8-((4-isopropyl- 395.15 1 H NMR (400 MHz, DMSO- phenyl)amino)-3- p [M+H]+ d6) 6 (ppm): 9.50 - 9.56 (m,
[1395] K • z
[1396] methyl-6-(2-methyl- ( ''2•- W / 1 H), 8.22 - 8.24 (m, 1 H), 7.79 morpholino)pyrimido[5, A 2- * - 7.81 (m, 2H), 7.22 - 7.24 (m, 4-d]pyrimidin-4(3H)- 2H), 4.30 - 4.56 (m, 2H), 3.88 one - 3.90 (m, 1 H), 3.46 - 3.51 (m,
[1397] 5H), 2.97- 3.05 (m, 1 H), 2.87 - 2.96 (m, 1 H), 2.62 - 2.70 (m, 1 H), 1.13 - 1.30 (m, 9H). 39 55 3-ethyl-8-((4- 395.10 1 H NMR (300 MHz, DMSO- isopropylphenyl)amino) [M+H]+ d6) 6: 9.69 (s, 1 H), 8.31 (s, -6-morpholinopyrimido- 1 H), 7.88 - 7.95 (m, 2H), 7.22
[1398] '" ANH
[1399] [5,4-d]pyrimidin-4(3H)- frV’V, - 7.25 (m, 2H), 4.02 - 4.06 (m, one x-x N A N N ' 2H), 3.63 - 4.04 (m, 8H), 2.87
[1400] 6,v. J 6
[1401] - 2.90 (m, 1 H), 1.21 - 1.29 (m, 9H).
[1402] 40 56 (R)-3-cyclopropyl-8-((4- 469.15 1 H NMR (400 MHz, DMSO- (3,3-difluorocyclobutyl)- [M+H]+ d6) 6 (ppm): 9.55 (s, 1 H), phenyl)amino)-6-(2- 8.05 (s, 1 H), 7.84 - 7.86 (m, methylmorpholino)pyri F, 2H), 7.29 - 7.31 (m, 2H), 4.40 mido[5,4-d]pyrimidin- - 4.52 (m, 2H), 3.89 - 3.93 (m, 4(3H)-one 1 H), 3.46 - 3.53 (m, 2H), 3.36
[1403] 6 o - 3.40 (m, 1 H), 3.21 - 3.25 (m, 1
[1404] 1 H), 2.93 - 3.03 (m, 3H), 2.61 - 2.74 (m, 3H), 1.16 - 1.17 (m, 3H), 1.03 - 1.08 (m, 2H), 0.90 - 0.94 (m, 2H).
[1405]
[1406] PATENT
[1407] ATTORNEY-DOCKET NO.: 51478-036WO4
[1408] 41 57 (R)-3-ethyl-6-(2- 436.10 1H NMR (300 MHz, DMSO) 6 methylmorpholino)-8- [M+H]+ (ppm): 9.33 (s, 1H), 8.75 - ((5-(trifluoromethyl)- 8.78 (m, 1H), 8.45 - 8.52 (m, pyridin-2-yl)amino)- 1H), 8.30 - 8.35 (m, 2H), 4.46 pyrimido[5,4- A - 4.54 (m, 2H), 3.94 - 4.06 (m, d]pyrimidin-4(3H)-one i 3H), 3.52 - 3.60 (m, 2H), 3.04 - 3.10 (m, 1H), 2.69 - 2.78 (m, 1H), 1.26 - 1.37 (m, 3H), 1.19 - 1.25 (m, 3H).
[1409] 42 58 (R)-3-ethyl-6-(2- 436.10 1H NMR (300 MHz, DMSO) 6 methylmorpholino)-8- [M+H]+ (ppm): 10.24 (s, 1H), 9.22 - ((6-(trifluoromethyl)- 9.28 (m, 1H), 8.55 - 8.64 (m, pyridin-3-yl)amino)- fsC 1H), 8.29 (s, 1H), 7.88 - 7.96 'C! L
[1410] pyrimido[5,4- A _ (m, 1H), 4.38 - 4.50 (m, 2H), Y i
[1411] d]pyrimidin-4(3H)-one 3.85 - 4.05 (m, 3H), 3.42 - 6 J o
[1412] I 3.56 (m, 2H), 2.97 - 3.08 (m,
[1413] 1H), 2.61 - 2.74 (m, 1H), 1.23 - 1.36 (m, 3H), 1.14 - 1.22 (m, 3H).
[1414] 43 59 (R)-3-cyclopropyl-8-((4- 421.25 1H NMR (500 MHz, MeOD) 6 isopropylphenyl)amino) [M+H]+ 8.32 (s, 1H), 7.64 - 7.69 (m, -6-(2-methyl- 2H), 7.32 - 7.37 (m, 2H), 4.40 morpholino)pyrimido[5, - 4.50 (m, 2H), 4.00 - 4.03 (m,
[1415] ''' NM
[1416] 4-d]pyrimidin-4(3H)-, A -N-, 1H), 3.61 - 3.72 (m, 2H), 3.34.. *!■
[1417] < N 'JI’ Y N
[1418] one a.. J 6 ''' - 3.41 (m, 1H), 3.21 - 3.31 (m,
[1419] I
[1420] 1H), 2.88 - 3.03 (m, 2H), 1.29 - 1.33 (m, 6H), 1.25 - 1.27 (m, 5H), 1.02 - 1.09 (m, 2H).
[1421]
[1422] Example 44: 6-(diethylamino)-3-ethyl-8-((4-isopropylphenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 22)
[1423] Step 1. Synthesis of 6-(diethylamino)-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one " NH
[1424] ''y "'o
[1425] N- %Ei0H,N
[1426] [i I: N
[1427] .. AA. N. - *• |i M
[1428] C N'n' 1 80'C, 1 hr >
[1429]
[1430] ° >0 1
[1431] To a stirred solution of 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d]pyrimidin-4(3H)-one (260 mg, 1.080 mmol, 1 equiv) in EtOH (4 mL) was added diethylamine (395.10 mg, 5.400 mmol, 5 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1h under nitrogen atmosphere. LCMS showed the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatographyPATENT
[1432] ATTORNEY-DOCKET NO.: 51478-036WO4
[1433] with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 6-(diethylamino)-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (280 mg, 93.45% yield, 98% purity) as a yellow oil. LC / MS: MS (ESI) calcd. for C13H19N5O2: 277.15. Found: 278.10 [M+H]+.
[1434] Step 2. Synthesis of 6-(diethylamino)-3-ethyl-8-hydroxypyrimido[5,4-d]pyrimidin-4(3H)-one
[1435] ”0 OH
[1436] LiCI, TsOH, DMF
[1437] *►
[1438] 120X, 1 hr
[1439]
[1440] To a stirred solution of 6-(diethylamino)-3-ethyl-8-methoxypyrimido[5,4-d]pyrimidin-4(3H)-one (280 mg, 1.010 mmol, 1 equiv) and LiCI (213.99 mg, 5.050 mmol, 5 equiv) in DMSO (4 mL) was added TsOH (869.30 mg, 5.050 mmol, 5 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 120 °C for 1h under nitrogen atmosphere. LCMS showed the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 6-(diethylamino)-3-ethyl-8-hydroxypyrimido[5,4-d]pyrimidin-4(3H)-one (260 mg, 97.80% yield, 92% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C12H17N5O2: 263.14. Found: 264.10 [M+H]+
[1441] Step 3. Synthesis of 8-chloro-6-(diethylamino)-3-ethylpyrimido[5,4-d]pyrimidin-4(3H)-one
[1442] OH Cl jSS.P0CI-. _ M
[1443] A -A N._
[1444] NNH ) BOX, 1 hr " N N Tf -i
[1445]
[1446] J O1O '
[1447] To a stirred solution of 6-(diethylamino)-3-ethyl-8-hydroxypyrimido[5,4-d]pyrimidin-4(3H)-one (120 mg, 0.456 mmol, 1 equiv) in phosphoryl trichloride (0.5 mL) was added phosphoryl trichloride (3.5 mL) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1h under nitrogen atmosphere. After completion of reaction, the resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 7 with saturated NaHCCh (aq.). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product, which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 8-chloro-6-(diethylamino)-3-ethylpyrimido[5,4-d]pyrimidin-4(3H)-one (120 mg, 93.46% yield, 97% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C12H16CIN5O: 281.10. Found: 282.10 [M+H]+PATENT
[1448] ATTORNEY-DOCKET NO.: 51478-036WO4
[1449] Step 4. Synthesis of 6-(diethylamino)-3-ethyl-8-((4-isopropylphenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1450] M W D1EA, DMSONAx.,
[1451] •'-'n, N, ” xx A A N
[1452] '■ N N V i 1D0‘C, 16hrs " W N" V >
[1453]
[1454] > O ' > O '
[1455] To a stirred solution of 8-chloro-6-(diethylamino)-3-ethylpyrimido[5,4-d]pyrimidin-4(3H)-one (120 mg, 0.426 mmol, 1 equiv) and para-isopropylaniline (69.11 mg, 0.511 mmol, 1.2 equiv) in DMSO (3 mL) was added DIEA (110.10 mg, 0.852 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1h under nitrogen atmosphere. After completion of reaction, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 6-(diethylamino)-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (78.7 mg, 48.56% yield, 99.9% purity) as a white solid. LC / MS: MS (ESI) calcd. for C21H28N6O: 380.23. Found: 381.15 [M+H]+ 1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 9.34 (s, 1H), 8.20 (s, 1H), 7.85 -7.92 (m, 2H), 7.20 - 7.23 (m, 2H), 3.99 - 4.01 (m, 2H), 3.63 - 3.65 (m, 4H), 2.82 - 2.87 (m, 1 H), 1.29 - 1.35 (m, 3H), 1.15 - 1.21 (m, 12H)
[1456] Using either 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d]pyrimidin-4(3H)-one or 6-chloro-3-isopropyl-8-methoxypyrimido[5,4-d]pyrimidin-4(3H)-one, the compounds of Table 3 were synthesized starting from the corresponding amines.
[1457] Table 3: Experimental Data for Compounds
[1458] Ex. Cmpd
[1459] Name Structure MS 1H NMR
[1460] # #
[1461] 45 23 6-(8-oxa-3- 421.15 1H NMR (400 MHz, DMSO-d6) azabicyclo[3.2.1] [M+H]+6 (ppm): 9.49 - 9.81 (m, 1H), octan-3-yl)-3- 8.19 - 8.42 (m, 1 H), 7.69 - 7.99 ethyl-8-((4- (m, 2H), 7.12 - 7.41 (m, 2H),
[1462] Ari
[1463] isopropylphenyl) NH 4.41 - 4.51 (m, 2H), 4.19 - 4.29 amino)pyrimido[ (m, 2H), 3.99 - 4.12 (m, 2H), 5,4-d]pyrimidin- SNN 3.17 - 3.21 (m, 2H), 2.72 - 3.00 0f
[1464] 4(3H)-one (m, 1H), 1.72 - 1.91 (m, 2H),
[1465] 1.57 - 1.72 (m, 2H), 1.35 - 1.22 (m, 3H), 1.09 - 1.22 (m, 6H).
[1466]
[1467] PATENT
[1468] ATTORNEY-DOCKET NO.: 51478-036WO4
[1469] 46 24 (S)-3-isopropyl- 423.20 1 H NMR (300 MHz, DMSO-d6)
[1470] 8-((4- [M+H]+ 6 (ppm): 9.42 - 9.43 (m, 1 H), isopropylphenyl) 8.21 - 8.22 (m, 1 H), 7.77 - 7.80 amino)-6-(2- (m, 2H), 7.22 - 7.25 (m, 2H), methylmorpholin 4.91 - 5.00 (m, 1 H), 4.45 - 4.53 o)pyrimido[5,4- (m, 2H), 3.89 - 3.94 (m, 1 H), d]pyrimidin- 3.44 - 3.54 (m, 2H), 2.82 - 3.02 4(3H)-one (m, 2H), 2.60 - 2.70 (m, 1 H),
[1471] 1.35 - 1.57 (m, 6H), 1.21 - 1.30 X
[1472] ') Q - (m, 9H).
[1473] 47 25 3-ethyl-8-((4- ( }.
[1474] V Vz- 407.15[ 1 H NMR (300 MHz, DMSO-d6) isopropylphenyl) )z=
[1475] Y zz- M+H]+ 6: 10.21 (s, 1 H), 8.43 - 8.44 (m, Zzx _
[1476] amino)-6-(2-oxa- 1 H), 7.88 - 7.89 (m, 2H), 7.28 - 6- 7.30 (m, 2H), 4.73 - 4.75 (m, NH f _ X
[1477] azaspiro[3.3]hep X 4H), 4.38 - 4.40 (m, 4H), 4.05 - N
[1478] tan-6- A -A N 4.09 (m, 2H), 2.90 - 2.97 (m, yl)pyrimido[5,4-N'Y1
[1479] dJ"J 0 11 H), 1.30 - 1.33 (m, 3H), 1.22 - d]pyrimidin- 1.28 (m, 6H).
[1480] 4(3H)-one
[1481]
[1482] In a similar approach to the one used in Example 44, using either 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d]pyrimidin-4(3H)-one, the compounds of Table 3A were synthesized starting from the corresponding amines.
[1483] Table 3A: Experimental Data for Compounds
[1484] Ex Cmpd
[1485] Name Structure MS 1H NMR
[1486] # #
[1487] 48 83 6-((1 S,4S)-2-oxa-5- 407.25 1 H NMR (500 MHz, MeOD) 6 azabicyclo[2.2.1]heptan- [M+H]+ 8.36 (s, 1 H), 7.75 - 7.77 (m, 5-yl)-3-ethyl-8-((4- 2H), 7.30 - 7.34 (m, 2H), 5.19 isopropyl- (s, 1 H), 4.79 - 4.82 (m, 1 H), phenyl)amino)pyrimido[5 4.14 - 4.19 (m, 2H), 3.94 (s,,4-d]pyrimidin-4(3H)-one 2H), 3.68 - 3.76 (m, 2H), 2.93
[1488] - 2.98 (m, 1 H), 2.08 (s, 2H), 1.42 - 1.45 (m, 3H), 1.29 - 1.33 (m, 6H).
[1489]
[1490] PATENT
[1491] ATTORNEY-DOCKET NO.: 51478-036WO4
[1492] 49 84 (R)-3-ethyl-8-((4- 409.25 1 H NMR (500 MHz, MeOD):
[1493] isopropylphenyl)amino)- [M+H]+ 6 8.39 (s, 1 H), 7.68 - 7.70 (m, 6-(3-methylmorpholino)- 2H), 7.33 - 7.35 (m, 2H), 4.69 pyrimido[5,4-d]pyrimidin- - 4.71 (m, 1 H), 4.24 - 4.26 (m, 4(3H)-one 1 H), 4.15 - 4.18 (m, 2H), 4.02
[1494] - 4.05 (m, 1 H), 3.83 - 3.85 (m, 1 H), 3.73 - 3.75 (m, 1 H), 3.52 - 3.58 (m, 1 H), 3.48 - 3.50 (m, 1 H), 2.90 - 3.02 (m, 1 H), 1.40 - 1.48 (m, 6H), 1.27 - 1.30 (m, 6H).
[1495] 50 85 (S)-3-ethyl-8-((4- 409.25 1 H NMR (500 MHz, MeOD):
[1496] isopropylphenyl)amino)- [M+H]+ 6 8.37 (s, 1 H), 7.68 - 7.70 (m, 6-(3-methylmorpholino)- 2H), 7.31 - 7.37 (m, 2H), 4.67 pyrimido[5,4-d]pyrimidin- - 4.75 (m, 1 H), 4.23 - 4.30 (m, 4(3H)-one 1 H), 4.13 - 4.21 (m, 2H), 3.99
[1497] - 4.07 (m, 1 H), 3.84 - 3.86 (m, 1 H), 3.74 - 3.77 (m, 1 H), 3.56 - 3.67 (m, 1 H), 3.47 - 3.55 (m, 1 H), 2.90 - 3.02 (m, 1 H), 1.39 - 1.48 (m, 6H), 1.29 - 1.32 (m,6H).
[1498] 51 86 6-((1 S,4R)-2-oxa-5- 407.25 1 H NMR (500 MHz, MeOD) 6 azabicyclo[2.2.1]heptan- [M+H]+ 8.46 (s, 1 H), 7.71 - 7.74 (m, 5-yl)-3-ethyl-8-((4- 2H), 7.35 - 7.37 (m, 2H), 5.18 isopropylphenyl)amino)p - 5.25 (m, 1 H), 4.83 (s, 1 H), yrimido[5,4-d]pyrimidin- 4.15 - 4.23 (m, 2H), 3.92 - »t4 A
[1499] 4(3H)-one 4.00 (m, 2H), 3.78 - 3.82 (m,
[1500] 0 D 1 H), 3.70 - 3.72 (m, 1 H), 2.94
[1501] - 3.00 (m, 1 H), 2.1 1 (s, 2H), 1.42 - 1.45 (m, 3H), 1.28 - 1.34 (m, 6H).
[1502]
[1503] PATENT
[1504] ATTORNEY-DOCKET NO.: 51478-036WO4
[1505] Example 52: 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-((4-isopropylphenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 26)
[1506] Step 1. Synthesis of 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one
[1507] OH
[1508] D
[1509] rY 'on
[1510] N ' Y. Pd(cippf}C!2. KJPOJ, dioxane / HjO
[1511] A. Y. - * JI 1 i
[1512] Cl8O‘C, 2 hrs sAA"' 'N'xY >
[1513] 0' N 6
[1514]
[1515] To a solution of 6-chloro-3-ethyl-8-methoxy- [1,3] diazino[5,4-d] pyrimidin-4-one (300 mg, 1.247 mmol, 1 equiv), 2-cyclopropylpyridin-4-ylboronic acid (243.81 mg, 1.496 mmol, 1.2 equiv), Pd(dppf)Cl2 (91.22 mg, 0.125 mmol, 0.1 equiv) and K2CO3 (344.58 mg, 2.494 mmol, 2 equiv) in Dioxane (5 mL) was added H2O (1 mL) in portions at room temperature under nitrogen atmosphere, he resulting mixture was stirred at 80 °C for 2h under nitrogen atmosphere, he resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. his resulted in 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (400 mg, 99.23% yield, 90% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C17H17N5O2: 323.14. Found: 324.20 [M+H]+
[1516] Step 2. Synthesis of 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-hydroxypyrimido[5,4-d] pyrimidin-4(3H)-one
[1517] o OH
[1518]
[1519] To a solution of 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-methoxy- [1,3] diazino[5,4-d] pyrimidin-4-one (200 mg, 0.619 mmol, 1 equiv) and LiCI (131.09 mg, 3.095 mmol, 5 equiv) in DMSO (4 mL) was added TsOH (532.54 mg, 3.095 mmol, 5 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 120 °C for 1h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-hydroxypyrimido[5,4-d] pyrimidin-4(3H)-one (180 mg, 94.08% yield, 96% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C16H15N5O2: 309.12. Found: 310.10 [M+H]+PATENT
[1520] ATTORNEY-DOCKET NO.: 51478-036WO4
[1521] Step 3. Synthesis of 8-chloro-6-(2-cyclopropylpyridin-4-yl)-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one
[1522]
[1523] To a solution of 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-hydroxypyrimido[5,4-d] pyrimidin-4(3H)-one (180 mg, 0.582 mmol, 1 equiv) in phosphoryl trichloride (1 mL) was added phosphoryl trichloride (4 mL) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1h under nitrogen atmosphere. The reaction was quenched with ice water at 0°C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 8-chloro-6-(2-cyclopropylpyridin-4-yl)-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one (130 mg, 68.16% yield, 97% purity) as a white solid. LC / MS: MS (ESI) calcd. for Ci6Hi4CIN5O:327.09. Found:328.05 [M+H]+
[1524] Step 4. Synthesis of 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-((4-isopropylphenyl)amino)-pyrimido[5,4-d]pyrimidin-4(3H)-one
[1525]
[1526] To a solution of 8-chloro-6-(2-cyclopropylpyridin-4-yl)-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.309 mmol, 1 equiv) and 1-isopropylpyrazol-4-amine (46.39 mg, 0.371 mmol, 1.2 equiv) in DMSO (1 mL) was added DIEA (79.84 mg, 0.618 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100 °C for 1h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 6-(2-cyclopropylpyridin-4-yl)-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (80.8 mg, 49.69% yield, 99.7% purity) as a trifluoroacetate salt and as a white solid. LC / MS: MS (ESI) calcd. for C25H26N6O:
[1527] 426.22. Found: 427.15 [M+H]+ 1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 10.04 (s, 1H), 8.63 - 8.64 (m, 2H), 8.06 - 8.14 (m, 2H), 7.92 - 7.94 (m, 2H), 7.31 - 7.34 (m, 2H), 4.07 - 4.09 (m, 2H), 2.91 - 2.96 (m, 1H), 2.27 - 2.32 (m, 1 H), 1.36 - 1.39 (m, 3H), 1.30 - 1.34 (m, 6H), 1.18 - 1.25 (m, 2H), 1.00 - 1.04 (m, 2H).PATENT
[1528] ATTORNEY-DOCKET NO.: 51478-036WO4
[1529] Example 53: (R)-3-ethyl-8-((4-isopropylphenyl)amino)-2-methyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 7)
[1530] Step 1. Synthesis of 6-chloro-3-ethyl-8-methoxy-2-methylpyrimido[5,4-d] pyrimidin-4(3H)-one
[1531]
[1532] To a stirred solution of 5-amino-2-chloro-N-ethyl-6-methoxypyrimidine-4-carboxamide (800 mg, 3.468 mmol, 1 equiv) in 1,1,1 -triethoxyethane (8 mL) was added acetic acid (624.86 mg, 10.404 mmol, 3 equiv) at room temperature. The reaction mixture was stirred at 120 °C for a period of 5 h. After completion of reaction, the reaction mixture was quenched by addition of water 25 mL. The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by column chromatography using 5% to 80% ethyl acetate in PE gradient to afford desired compound 6-chloro-3-ethyl-8-methoxy-2-methylpyrimido[5,4-d] pyrimidin-4(3H)-one (780 mg, 88.30% yield, 95% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for CIOHHCIN402:254.06, Found: 255.20.
[1533] Step 2. Synthesis of (R)-3-ethyl-8-methoxy-2-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1534]
[1535] To a stirred solution of 6-chloro-3-ethyl-8-methoxy-2-methyl- [1,3] diazino[5,4-d] pyrimidin-4-one (200 mg, 0.785 mmol, 1 equiv) in anhydrous DMSO (2 mL) were added (2R)-2-methylmorpholine (119.15 mg, 1.177 mmol, 1.5 equiv) and DIEA (203.00 mg, 1.570 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 100 °C for a period of 2 h. After completion of reaction, the reaction mixture was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nmto afford desired compound (R)-3-ethyl-8-methoxy-2-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (190 mg, 75.76% yield, 93% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H21 N5O3: 319.16, Found: 320.20.PATENT
[1536] ATTORNEY-DOCKET NO.: 51478-036WO4
[1537] Step 3. Synthesis of (R)-3-ethyl-8-hydroxy-2-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1538] O OH
[1539] LiCI, TsOH, DMF
[1540] 120’C, 1 hr
[1541]
[1542] To a stirred solution of 3-ethyl-8-methoxy-2-methyl-6-[(2R)-2-methylmorpholin-4-yl]- [1,3] diazino[5,4-d] pyrimidin-4-one (170 mg, 0.532 mmol, 1 equiv) in anhydrous DMF (2 mL) were added LiCI (112.82 mg, 2.660 mmol, 5 equiv) and TsOH (458.32 mg, 2.660 mmol, 5 equiv) at room temperature. The reaction mixture was stirred at 120 °C for a period of 1 h. After completion of reaction, the reaction mixture was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford desired compound (R)-3-ethyl-8-hydroxy-2-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (140 mg, 86.14% yield, 93% purity) as a white solid. LC / MS: MS (ESI) calcd. for C14H19N5O3: 305.15, Found: 306.20.
[1543] Step 4. Synthesis of (R)-8-chloro-3-ethyl-2-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1544]
[1545] A mixture of 3-ethyl-8-hydroxy-2-methyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (120 mg, 0.393 mmol, 1 equiv) in POCl3 (2 mL) was stirred for 1 h at 80 °C. After completion of reaction, the resulting mixture was concentrated under reduced pressure. The residue was basified to pH = 8 with saturated NaHCCh (aq.). The aqueous layer was extracted with EtOAc (3x 25 mL). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product. The resulting mixture was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-8-chloro-3-ethyl-2-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (98 mg, 77.02% yield, 93% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C14H18CIN5O2: 323.11, Found: 324.20.PATENT
[1546] ATTORNEY-DOCKET NO.: 51478-036WO4
[1547] Step 5. Synthesis of (R)-3-ethyl-8-((4-isopropylphenyl)amino)-2-methyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1548]
[1549] To a stirred solution of 8-chloro-3-ethyl-2-methyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (95 mg, 0.293 mmol, 1 equiv) in anhydrous DMSO (2 mL) were added paraisopropylaniline (47.61 mg, 0.352 mmol, 1.2 equiv) and DIEA (75.84 mg, 0.586 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 100 °C for a period of 2 h. After completion of reaction, the reaction mixture was quenched by addition of water 25 mL. The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was washed with brine (2 x 25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford desired compound (R)-3-ethyl-8-((4-isopropylphenyl)amino)-2-methyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (55 mg, 44.36% yield, 99.8% purity) as a light yellow solid. LC / MS: MS (ESI) calcd. for C23H3oN602:422.24, Found: 423.25.1H NMR (300 MHz, DMSO-d6) 6 (ppm): 9.48 - 9.49 (m, 1H), 7.75 - 7.79 (m, 2H), 7.24 - 7.28 (m, 2H), 4.45 - 4.50 (m, 2H), 4.03 - 4.10 (m, 2H), 3.89 - 3.94 (m, 1 H), 3.45 - 3.56 (m, 2H), 2.84 - 3.06 (m, 2H), 2.65 - 2.72 (m, 4H), 1.01 - 1.32 (m,12H).
[1550] Example 54: (R)-3-(2-(dimethylamino)ethyl)-8-((4-isopropylphenyl)amino)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 4)
[1551] Step 1. Synthesis of 5-amino-2-chloro-N-(2-(dimethylamino) ethyl)-6-methoxypyrimidine-4-carboxamide
[1552] HZN..
[1553] "'O "" O
[1554] ... NH2HATU, DIEA. DMF K>4^X, NH2
[1555] I i _OH - ► n s.
[1556] Cl N rt, 1 hr Cl' N' y '
[1557] o o
[1558]
[1559] To a solution of 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid (1 g, 4.912 mmol, 1 equiv), (2-aminoethyl) dimethylamine (649.53 mg, 7.368 mmol, 1.5 equiv) and HATU (3735.48 mg, 9.824 mmol, 2 equiv) in DMF (5 mL) was added DIEA (1904.61 mg, 14.736 mmol, 3.00 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2h under nitrogen atmosphere. The resulting mixture was extracted with EtOAc (3 x 80 mL). The combined organic layers were washed with water (3 x 70 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gelPATENT
[1560] ATTORNEY-DOCKET NO.: 51478-036WO4
[1561] column chromatography, eluted with CH2Cl2 / MeOH (10:1) to afford 5-amino-2-chloro-N-(2-(dimethylamino) ethyl)-6-methoxypyrimidine-4-carboxamide (1.1 g, 81.81% yield, 98% purity) as a white solid. LC / MS: MS (ESI) calcd. for C10H16CIN5O2: 273.10. Found: 274.10 [M+H]+
[1562] Step 2. Synthesis of 6-chloro-3-(2-(dimethylamino) ethyl)-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one
[1563]
[1564] To a solution of 5-amino-2-chloro-N-[2-(dimethylamino) ethyl]-6-methoxypyrimidine-4-carboxamide (500 mg, 1.827 mmol, 1 equiv) in trimethoxymethane (4.5 mL) was added HOAc (0.5 mL) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 120 °C for 16 h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in 6-chloro-3-(2-(dimethylamino) ethyl)-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (350 mg, 67.53% yield, 91% purity) as a white solid. LC / MS: MS (ESI) calcd. for C11H14CIN5O2: 283.08. Found: 284.10 [M+H]+.
[1565] Step 3. Synthesis of (R)-3-(2-(dimethylamino) ethyl)-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1566] BO'C, 1 hr
[1567]
[1568] To a solution of 6-chloro-3-(2-(dimethylamino) ethyl)-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (350 mg, 1.234 mmol, 1 equiv) and (2R)-2-methylmorpholine (149.73 mg, 1.481 mmol, 1.2 equiv) in DMSO (5 mL) was added DIEA (318.88 mg, 2.468 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1 h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-3-(2-(dimethylamino) ethyl)-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (400 mg, 93.07% yield, 91% purity) as a white solid. LC / MS: MS (ESI) calcd. for C16H24N6O3: 348.19. Found: 349.20 [M+H]+.PATENT
[1569] ATTORNEY-DOCKET NO.: 51478-036WO4
[1570] Step 4. Synthesis of (R)-3-(2-(dimethylamino) ethyl)-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1571] '■'■Q OH
[1572] ■AA LiCI, TSOH, DMF
[1573] ...-x.. X. ANIN
[1574] r N N If > 120‘C, 1 hr f N 'N V >
[1575] 0M0V
[1576]
[1577] To a solution of (R)-3-(2-(dimethylamino) ethyl)-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (200 mg, 0.574 mmol, 1 equiv) and LiCI (121.67 mg, 2.870 mmol, 5 equiv) in DMSO (4 mL) was added TsOH (494.25 mg, 2.870 mmol, 5 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 120 °C for 1h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-3-(2-(dimethylamino) ethyl)-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (70 mg, 36.47% yield, 96% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H22N6O3: 334.18. Found:
[1578] 335.20 [M+H]+.
[1579] Step 5. Synthesis of (R)-8-chloro-3-(2-(dimethylamino) ethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1580] POCI3 80 °C, 1 hr
[1581]
[1582] A solution of (R)-3-(2-(dimethylamino) ethyl)-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 0.179 mmol, 1 equiv) in phosphoryl trichloride (0.5 mL) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 1h under nitrogen atmosphere. The reaction was quenched with ice water at 0°C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-8-chloro-3-(2-(dimethylamino) ethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 94.77% yield, 96% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C15H21CIN6O2: 352.14. Found: 353.20 [M+H]+.PATENT
[1583] ATTORNEY-DOCKET NO.: 51478-036WO4
[1584] Step 6. Synthesis of (R)-3-(2-(dimethylamino)ethyl)-8-((4-isopropylphenyl)amino)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1585]
[1586] To a solution of (R)-8-chloro-3-(2-(dimethylamino) ethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 0.170 mmol, 1 equiv) and para-isopropylaniline (27.59 mg, 0.204 mmol, 1.2 equiv) in DMSO (3 mL) was added DIEA (43.96 mg, 0.340 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100 °C for 1h under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254nm. This resulted in (R)-3-(2-(dimethylamino) ethyl)-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one; trifluoroacetic acid salt; (14.1 mg, 14.66% yield, 99.0% purity) as a white solid. LC / MS: MS (ESI) calcd. for C24H33N7O2: 451.27. Found: 452.25 [M+H]+ 1H NMR (400 MHz, DMSO-c / e) 6 (ppm): 8.90 - 8.94 (m, 1H), 8.21 (s, 1H), 7.76 - 7.82 (m, 2H), 7.24 - 7.26 (m, 2H), 4.51 - 4.54 (m, 2H), 4.42 - 4.45 (m, 2H), 3.93 - 3.94 (m, 1H), 3.43 - 3.52 (m, 4H), 3.17 - 3.18 (m, 1H), 2.95 - 3.07 (m, 1H), 2.88 -2.98 (m, 6H), 2.52 - 2.61 (m, 1H), 1.19 - 1.21 (m, 9H).
[1587] Using the same approach, the compound of Table 4 was synthesized from the corresponding amine and 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid.
[1588] Table 4: Experimental Data for Compounds
[1589] Ex. Cmpd
[1590] Name Structure MS 1H NMR
[1591] # #
[1592] 55 27 (R)-8-((4- 478.35 1H NMR (500 MHz, MeOD) 6 isopropylphenyl)- [M+H]+ 9.40 - 9.43 (m, 1H), 8.19 (s, amino)-6-(2- 1H), 7.68 - 7.70 (m, 2H), 7.30 methylmorpholino)-3- - 7.31 (m, 2H), 4.54 - 4.62 (m, (1-methylpiperidin-4- 2H), 3.99 - 4.01 (m, 1H), 3.72 yl)pyrimido[5,4- - 3.75 (m, 2H), 3.64 - 3.66 (m, d]pyrimidin-4(3H)-one 2H), 3.50 - 3.52 (m, 1H), 3.30 - 3.33 (m, 1H), 3.13 - 3.19 (m, 1H), 2.97 - 3.00 (m, 4H), 2.92 - 2.94 (m, 1H), 2.79 - 2.84 (m, 3H), 2.29 - 2.31 (m, 2H), 1.25 - 1.30 (m, 9H).
[1593]
[1594] PATENT
[1595] ATTORNEY-DOCKET NO.: 51478-036WO4
[1596] Example 56: 3-ethyl-8-((4-isopropylphenyl)amino)-6-(tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 28)
[1597] Step 1. Synthesis of 3-ethyl-8-methoxy-6-(tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1598] ,-~-x, Br
[1599] f T
[1600] ?wlr(dF(CF3)ppyl2(dto^>y)PFB, Y
[1601] N '■ Y, NiCI2(dtbbppy), TTMSS, N2CO3, DME€, X, Nc,N.... - i X
[1602] il i Blue LEDs, 16 hrs f Y N y 1
[1603]
[1604] ° 01
[1605] To a stirred solution of 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (200 mg, 0.831 mmol, 1 equiv) in anhydrous DME (5 mL) were added 4-bromotetrahydro-2H-pyran (205.73 mg, 1.246 mmol, 1.5 equiv), lr[dF(CF3) PPyHdtbpy)) PFe (9.32 mg, 0.008 mmol, 0.01 equiv), NiCl2(dtbbpy) (1.65 mg, 0.004 mmol, 0.005 equiv), TTMSS (219.96 mg, 0.831 mmol, 1 equiv) and Na2COs (176.17 mg, 1.662 mmol, 2 equiv) at room temperature under N2 atmosphere. The reaction mixture was stirred at room temperature for a period of 16 h under blue light irradiation. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL). The aqueous layer was extracted with ethyl acetate (3 x 25 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford 3-ethyl-8-methoxy-6-(tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (86 mg, 35.64% yield, 95% purity) as a light yellow solid. LC / MS: MS (ESI) calcd. for C14H18N4O3: 290.14, Found: 291.20.
[1606] Step 2. Synthesis of 3-ethyl-8-hydroxy-6-(tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1607] Y OH
[1608] N '[ ''i LiCI, TsOH, DMFKN,. X
[1609] A, A A,. * A J fNT| ] 120" C, 1 hr •' V N’ y >
[1610]
[1611] o ' o ’
[1612] To a stirred solution of 3-ethyl-8-methoxy-6-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (76 mg, 0.262 mmol, 1 equiv) in anhydrous DMF (2 mL) were added LiCI (55.48 mg, 1.310 mmol, 5 equiv) and TsOH (225.39 mg, 1.310 mmol, 5 equiv) at room temperature. The reaction mixture was stirred at 120 °C for a period of 1 h. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL). The aqueous layer was extracted with ethyl acetate (3 x 25 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford 3-ethyl-8-hydroxy-6-(tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (60 mg, 82.96% yield, 94% purity) as a white solid. LC / MS: MS (ESI) calcd. for Ci3Hi6N4O3:276.12, Found: 277.20.PATENT
[1613] ATTORNEY-DOCKET NO.: 51478-036WO4
[1614] Step 3. Synthesis of 8-chloro-3-ethyl-6-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1615] OH Cl
[1616] POCk Nx
[1617] _: _ T "i
[1618] A -A N
[1619] 80’C, 1 hr r YNif:
[1620]
[1621] o.. > O ’
[1622] A solution of 3-ethyl-8-hydroxy-6-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (55 mg, 0.199 mmol, 1 equiv) in POCl3 (50 mL) was stirred at 80 °C for a period of 1 h. After completion of reaction, the reaction mixture was concentrated under reduced pressure. The residue was basified to pH 7 with saturated NaHCOs (aq.). The aqueous layer was extracted with ethyl acetate (3 x 25 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 8-chloro-3-ethyl-6-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (50 mg, 85.22% yield, 92% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C13H15CIN4O2: 294.09, Found: 295.10.
[1623] Step 4. Synthesis of 3-ethyl-8-((4-isopropylphenyl)amino)-6-(tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1624] XI X i
[1625] ClvNH2
[1626] NAy DSEA, DMSONAy A
[1627] j I. N *■.... Jil A N
[1628] f TN|f j 50'C, 2 hr < V N A
[1629]
[1630] To a stirred solution of 8-chloro-3-ethyl-6-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (50 mg, 0.170 mmol, 1 equiv) in anhydrous DMSO (2 mL) were added 4-isopropylaniline (34.41 mg, 0.255 mmol, 1.5 equiv) and DIEA (43.85 mg, 0.340 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 50 °C for a period of 2 h. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL). The aqueous layer was extracted with ethyl acetate (3 x25 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford 3-ethyl-8-((4-isopropylphenyl)amino)-6-(tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (58.1 mg, 87.04% yield, 98.7% purity) as a white solid. LC / MS: MS (ESI) calcd. for C22H27N5O2: 393.22, Found: 394.15.1H NMR (300 MHz, DMSO-d6) 6 (ppm): 9.65 - 9.66 (m, 1 H), 8.53 - 8.54 (m, 1 H), 7.91 - 7.94 (m, 2H), 7.23 - 7.27 (m, 2H), 4.02 - 4.09 (m, 2H), 3.92 - 3.97 (m, 2H), 3.42 - 3.51 (m, 2H), 2.83 - 3.01 (m, 2H), 1.80 - 1.88 (m, 4H), 1.30 - 1.33 (m, 3H), 1.28 - 1.30 (m, 6H).PATENT
[1631] ATTORNEY-DOCKET NO.: 51478-036WO4
[1632] Example 57: (S)-6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl)amino)pyrimido-[5,4-d]pyrimidin-4(3H)-one and (R)-6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one - Stereochemistry arbitrarily assigned (Compound 29 and 30)
[1633] Step 1. Synthesis of 3-ethyl-6,8-dihydroxypyrimido[5,4-d] pyrimidin-4(3H)-one
[1634] OH
[1635] LtCf, TsOH, DMSO N " T' "'j
[1636] - ►. HO N
[1637] 120’C, 1 hr o I
[1638]
[1639] To a solution of 6-chloro-3-ethyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (1000 mg, 4.155 mmol, 1 equiv) and TsOH (3577.81 mg, 20.775 mmol, 5 equiv) in DMF (15 mL) was added LiCI (880.74 mg, 20.777 mmol, 5.00 equiv) at room temperature. The resulting mixture was stirred at 120°C for 1 h. Desired product could be detected by LCMS. The precipitated solids were collected by filtration and washed with water (3 x 10 mL). The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 30% gradient in 20 min; detector, UV 254 nm. This resulted in 3-ethyl-6,8-dihydroxypyrimido[5,4-d] pyrimidin-4(3H)-one (450 mg, 47.79% yield, 96% purity) as a white solid. MS: MS (ESI) calcd. for C8H8N4O3: 208.06, Found: 209.10 [M+H]+.
[1640] Step 2. Synthesis of 6,8-dichloro-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one
[1641] OH POCI3110”C, 1 hr
[1642]
[1643] O
[1644] To a mixture of 3-ethyl-6,8-dihydroxypyrimido[5,4-d] pyrimidin-4(3H)-one (450 mg, 2.162 mmol, 1 equiv) in POCl3 (5 mL) were added DIEA (1 mL) dropwise at room temperature. The resulting mixture was stirred at 110°C for 1 h. Desired product could be detected by LCMS. The resulting mixture was concentrated under vacuum. The mixture was neutralized to pH 8 with saturated NaHCCh (aq.). The resulting mixture was extracted with EtOAc (3 x 50mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in 6,8-dichloro-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one (400 mg, crude) as a brown solid. MS: MS (ESI) calcd. for C8H6CI2N4O: 243.99, Found: 245.00 [M+H]+.
[1645] Step 3. Synthesis of 6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1646] A
[1647]
[1648] 2. D16A, DMSO, 100‘ C. 4hrsPATENT
[1649] ATTORNEY-DOCKET NO.: 51478-036WO4
[1650] To a stirred solution of 6,8-dichloro-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one (120 mg, 0.490 mmol, 1 equiv) and 4-isopropylaniline (66.21 mg, 0.490 mmol, 1 equiv) in DMSO (2 mL) was added DIEA (126.58 mg, 0.980 mmol, 2 equiv) at room temperature. The resulting mixture was stirred at 50°C for 1 h. To the above mixture was added 2-cyclopropylmorpholine (74.74 mg, 0.588 mmol, 1.2 equiv) at room temperature. The resulting mixture was stirred at 100°C for 1h. Desired product could be detected by LCMS. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The resulting mixture was extracted with EtOAc (3 x 20 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 70% gradient in 30 min; detector, UV 254 nm. This resulted in 6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (90 mg, 42.30% yield, 96% purity) as a yellow solid. MS: MS (ESI) calcd. for C24H30N6O2: 434.24, Found: 435.15 [M+H]+.
[1651] Step 4. Separation of (R)-6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one and (S)-6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1652]
[1653] The product (90 mg) was purified by silica gel Column: CHIRALPAK IC, 5 pm, 250 mm x 20 mm; Mobile Phase A: MTBE (with 0.5% NH3 (2 M in MeOH)), Mobile Phase B: MEOH: DCM=1: 1; Flow rate: 20 mL / min; Gradient (B%): isocratic 7% B; to afford (R)-6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (23.7 mg, 31.25% yield, 99.5% purity) as a white solid. RT1(min): 14.67. LC / MS: MS (ESI) calcd. for C24H30N6O2: 432.24. Found: 435.15 [M+H]+ 1H NMR (400 MHz, DMSO+D2O) 6 (ppm): 9.89 - 9.90 (m, 2H), 8.25 (s, 1H), 7.75 - 7.77 (m, 2H), 7.23 - 7.25 (m, 2H), 4.56 - 4.59 (m, 1H), 4.39 - 4.42 (m, 1H), 3.91 - 4.03 (m, 3H), 3.39 - 3.44 (m, 1H), 3.00 - 3.06 (m, 1 H), 2.83 - 2.90 (m, 2H), 2.70 - 2.74 (m, 1 H), 1.27 - 1.30 (m, 3H), 1.20 - 1.22 (m, 6H), 0.89 - 0.92 (m, 1 H), 0.50 - 0.55 (m, 2H), 0.32 - 0.35 (m, 1H), 0.25 - 0.26 (m, 1H).
[1654] and (S)-6-(2-cyclopropylmorpholino)-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (24.3 mg, 32.04% yield, 99.9% purity) as an off-white solid. RT2(min): 19.78. LC / MS: MS (ESI) calcd. for C24H30N6O2: 434.24. Found: 435.15 [M+H]+1H NMR (400 MHz, DMSO-d6) 6 (ppm): 9.55 (s, 1 H), 8.25 (s, 1 H), 7.77 - 7.80 (m, 2H), 7.21 - 7.23 (m, 2H), 4.56 - 4.60 (m, 1 H), 4.37 - 4.41 (m, 1 H), 3.91 - 4.04 (m, 3H), 3.38 - 3.45 (m, 2H), 2.96 - 3.05 (m, 1 H), 2.71 - 2.92 (m, 2H), 1.25 - 1.30 (m, 3H), 1.19 - 1.22 (m, 6H), 0.88 - 0.92 (m, 1 H), 0.49 - 0.52 (m, 2H), 0.23 - 0.36 (m, 2H).
[1655] Stereochemistry was arbitrarily assigned.PATENT
[1656] ATTORNEY-DOCKET NO.: 51478-036WO4
[1657] Example 58: (R)-8-((1 -acetylpiperidin-4-yl)amino)-3-isopropyl-6-(2-methylmorpholino)pyrimido- [5,4-d]pyrimidin-4(3H)-one (Compound 31)
[1658] Step 1. Synthesis of (R)-8-((1-acetylpiperidin-4-yl)amino)-3-isopropyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1659]
[1660] A solution of (R)-8-chloro-3-isopropyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.309 mmol, 1 equiv), 1-(4-aminopiperidin-1-yl) ethan-1-one (48.31 mg, 0.340 mmol, 1.1 equiv), Pd2(dba)3 (28.28 mg, 0.031 mmol, 0.1 equiv), Xantphos (17.87 mg, 0.031 mmol, 0.1 equiv) and Cs₂CO₃ (301.89 mg, 0.927 mmol, 3 equiv) in Dioxane (2 mL) was stirred at 100°C for 1h under nitrogen atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-8-((1-acetylpiperidin-4-yl) amino)-3-isopropyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one; trifluoroacetic acid salt. (43.8 mg, 26.09% yield, 99.9% purity) as a light yellow semi-solid. LC / MS: MS (ESI) calcd. for C21H31N7O3: 429.25. Found: 430.20 [M+H]+ 1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 8.39 - 8.42 (m, 1 H), 7.59 - 7.72 (m, 1 H), 4.87 - 4.98 (m, 1H), 4.36 - 4.46 (m, 3H), 4.28 - 4.33 (m, 1H), 3.88 - 3.97 (m, 2H), 3.51 - 3.59 (m, 2H), 3.09 - 3.23 (m, 2H), 2.82 - 2.88 (m, 1 H), 2.64 - 2.76 (m, 1 H), 1.99 - 2.04 (m, 3H), 1.81 - 1.92 (m, 2H), 1.53 - 1.77 (m, 2H), 1.42 - 1.47 (m, 6H), 1.14 - 1.20 (m, 3H).
[1661] Using the same approach, the compounds of Table 5 were synthesized.
[1662] Table 5: Experimental Data for Compounds
[1663] Ex. Cmpd
[1664] Name Structure MS 1H NMR
[1665] # #
[1666] 59 6 (R)-3-isopropyl-8- 424.15 1H NMR (300 MHz, DMSO- ((5-isopropylpyridin- [M+H]+ d6) 6 (ppm): 9.36 - 9.40 (m, 2-yl)amino)-6-(2- 1H), 8.27 - 8.34 (m, 3H), 7.92 methylmorpholino)pNH- 7.95 (m, 1H), 4.96 - 5.00 (m, yrimido[5,4-.. X 1H), 4.43 - 4.54 (m, 2H), 3.93 d]pyrimidin-4(3H)- - 3.97 (m, 1H), 3.50 - 3.57 (m,
[1667] O. J O *
[1668] one 1 2H), 2.92 - 3.09 (m, 2H), 2.72 - 2.75 (m, 1H), 1.42 - 1.45 (m, 6H) 1.21 - 1.34 (m, 9H).
[1669]
[1670] PATENT
[1671] ATTORNEY-DOCKET NO.: 51478-036WO4
[1672] 60 5 (R)-3-isopropyl-8- 424.15 1 H NMR (300 MHz, DMSO- ((6-isopropylpyridin- [M+H]+ d6) 6 (ppm): 10.35 - 10.40 3-yl)amino)-6-(2- (m, 1 H), 9.26 - 9.32 (m, 1 H), methylmorpholino)p 8.76 - 8.79 (m, 1 H), 8.20 - yrimido[5,4- 8.27 (m, 1 H), 7.88 - 7.92 (m, d]pyrimidin-4(3H)- '" MH
[1673] X,, N, 1 H), 4.96 - 5.00 (m, 1 H), 4.37 one ~ A A A X - 4.49 (m, 2H), 3.91 - 3.95 (m,
[1674] ( N M Y •
[1675] G J 6 = 1 H), 3.48 - 3.55 (m, 2H), 3.21 I - 3.31 (m, 1 H), 2.70 - 3.07 (m,
[1676] 1 H), 2.55 - 2.66 (m, 1 H), 1.44 - 1.66 (m, 6H) 1.35 - 1.40 (m, 6H), 1.19 - 1.32 (m, 3H).
[1677]
[1678] In a similar approach to the one used for example 58, using 6,8-dichloro-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one or 6,8-dichloro-3-methylpyrimido[5,4-d] [1,3] diazin-4-one, the compounds of Table 5A were synthesized starting from the corresponding amines.
[1679] Table 5A: Experimental Data for Compounds
[1680] Ex Cmpd
[1681] Name Structure MS 1H NMR
[1682] # #
[1683] 61 60 6-(6-oxa-3- 407.15 1 H-NMR (300 MHz, DMSO- azabicyclo[3.1,1]hepta [M+H]+d6) 6 (ppm): 8.37 - 8.38 (m, n-3-yl)-3-ethyl-8-((4- 1 H), 7.89 - 7.92 (m, 2H), isopropylphenyl)amino) 7.25 - 7.28 (m, 2H), 4.72 - pyrimido[5,4- 4.75 (m, 2H), 3.95 - 4.09 A,, N
[1684] d]pyrimidin-4(3H)-one N Y " I (m, 4H), 3.63 - 3.82 (m, 2H), oJJ d 3.12 - 3.16 (m, 1 H), 2.86 - 2.91 (m, 1 H), 1.84 - 1.88 (m, 1 H), 1.31 - 1.35 (m, 3H), 1.19 - 1.29 (m, 6H).
[1685] 62 61 (R)-3-ethyl-8-((4- 439.15 1 H NMR (400 MHz, DMSO) isopropylphenyl)amino) [M+H]+6 9.85 (s, 1 H), 8.33 (s, 1 H), -6-(2- 7.75 - 7.78 (m, 2H), 7.22 - (methoxymethyl)morph 7.24 (m, 2H), 4.53 - 4.56 olino)pyrimido[5,4- (m, 1 H), 4.38 - 4.41 (m, 1 H), d]pyrimidin-4(3H)-one N N
[1686] Q.X„J ci 3.93 - 3.99 (m, 3H), 3.66 - 3.56 (m, 1 H), 3.55 - 3.42 (m, 2H), 3.40 - 3.33 (m, 1 H), 3.31 (s, 3H), 3.12 - 3.00 (m, 1 H), 2.94 - 2.77 (m, 2H),
[1687]
[1688] PATENT
[1689] ATTORNEY-DOCKET NO.: 51478-036WO4
[1690] 1.33 - 1.25 (m, 3H), 1.21 - 1.23 (m, 6H).
[1691] 63 62 3-ethyl-6-(3- 381.15 1 H NMR (300 MHz, DMSO- hydroxyazetidin-1 -yl)- [M+H]+d6) 6 (ppm): 8.47 - 8.54 (m, 8-((4- 1 H), 7.80 - 7.89 (m, 2H), isopropylphenyl)amino) 7.27 - 7.38 (m, 2H), 4.56 - pyrimido[5,4- 4.67 (m, 1 H), 4.42 - 4.54 d]pyrimidin-4(3H)-one (m, 2H), 4.01 - 4.18 (m, 4H),
[1692] 2.85 - 2.96 (m, 1 H), 1.30 - 1.41 (m, 3H), 1.15 - 1.26 (m, 6H).
[1693] 64 63 3-ethyl-8-((4- 397.15 1 H NMR (300 MHz, DMSO) isopropylphenyl)amino) [M+H]+6 (ppm): 8.45 - 8.57 (m, -6-((2- 1 H), 7.70 - 7.83 (m, 2H), methoxyethyl)(methyl)a 7.19 - 7.32 (m, 2H), 4.01 - mino)pyrimido[5,4- 'lTi 4.12 (m, 2H), 3.80 - 3.89 d]pyrimidin-4(3H)-one (m, 3H), 3.62 - 3.69 (m, 2H),
[1694] !Y
[1695] 6 3.25 - 3.32 (m, 2H), 3.16 - 3.25 (m, 3H), 2.85 - 2.94 (m, 1 H), 1.25 - 1.47 (m, 3H), 1.08 - 1.23 (m, 6H).
[1696] 65 64 3-ethyl-6-(3- 383.15 1 H NMR (400 MHz, DMSO- fluoroazetidin-1 -yl)-8- [M+H]+d6) 6 (ppm): 8.41 - 8.48 (m, ((4- 1 H), 7.71 - 7.80 (m, 2H), isopropylphenyl)amino) 7.23 - 7.32 (m, 2H), 5.25 - pyrimido[5,4- 5.73 (m, 1 H), 4.40 - 4.63 d]pyrimidin-4(3H)-one (m, 2H), 4.20 - 4.37 (m, 2H),
[1697] 3.98 - 4.11 (m, 2H), 2.81 - 2.98 (m, 1 H), 1.30 - 1.37 (m, 3H), 0.99 - 1.23 (m, 6H).
[1698] 66 65 (R)-3-ethyl-8-((4- 409.15 1 H NMR (300 MHz, DMSO- isopropylphenyl)amino) [M+H]+d6) 6 (ppm): 8.41 (s, 1 H), -6-(3- 7.88 - 7.91 (m, 2H), 7.26 - methoxypyrrolidin-1 - 7.29 (m, 2H), 4.02 - 4.09 Af!
[1699] yl)pyrimido[5,4- YA (m, 3H), 3.51 - 3.78 (m, 4H), d]pyrimidin-4(3H)-one 327 (s, 3H), 2.84 - 2.94 (m,
[1700] ' \. J
[1701] 1 H), 2.02 - 2.09 (m, 2H), 1.29 - 1.33 (m, 3H), 1.20 - 1.22 (m, 6H).
[1702]
[1703] PATENT
[1704] ATTORNEY-DOCKET NO.: 51478-036WO4
[1705] 67 66 (S)-3-ethyl-8-((4- 439.20 1 H NMR (400 MHz, DMSO- isopropylphenyl)amino) [M+H]+d6) 6 (ppm): 9.56 - 9.59 (m, -6-(2- 1 H), 8.28 - 8.32 (m, 1 H), (methoxymethyl)morph 7.78 - 7.80 (m, 2H), 7.21 - olino)pyrimido[5,4- 7.23 (m, 2H), 4.56 - 4.59 d]pyrimidin-4(3H)-one (m, 1 H), 4.37 - 4.48 (m, 1 H),
[1706] J- 3.92 - 4.04 (m, 3H), 3.58 - 3.63 (m, 1 H), 3.42 - 3.58 'o (m, 2H), 3.37 - 3.44 (m, 1 H),
[1707] 3.28 - 3.38 (m, 3H), 2.98 - 2.13 (m, 1 H), 2.88 - 2.98 (m, 1 H), 2.72 - 2.86 (m, 1 H), 1.26 - 1.32 (m, 3H), 1.18 - 1.28(m, 6H).
[1708] 68 67 (R)-3-ethyl-6-(3- 395.15 1 H NMR (400 MHz, DMSO) hydroxypyrrolidin-1-yl)- [M+H]+6 (ppm) 10.15 (s, 1 H), 8.42 8-((4- (s, 1 H), 7.70 - 8.08 (m, 2H), isopropylphenyl)amino) 7.10 - 7.39 (m, 2H), 4.41 (s, pyrimido[5,4- 1 H), 3.99 - 4.12 (m, 2H), d]pyrimidin-4(3H)-one 3.61 - 3.79 (m, 3H), 3.51 - HO. / '’*' ’M' ’r<’ ’
[1709] 3.61 (m, 1 H), 2.80 - 2.99 (m, 1 H), 1.98 - 2.10 (m, 1 H), 1.93 (s, 1 H), 1.27 - 1.37 (m, 3H), 1.13 - 1.37 (m, 6H). 69 68 (S)-3-ethyl-6-(3- 395.15 1 H NMR (300 MHz, DMSO- hydroxypyrrolidin-1-yl)- [M+H]+d6) 6 (ppm): 8.41 (s, 1 H), 8-((4- 7.89 - 7.91 (m, 2H), 7.26 - isopropylphenyl)amino) 7.29 (m, 2H), 4.30 - 4.40 pyrimido[5,4- (m, 1 H), 3.95 - 4.15 (m, 2H),
[1710] , N,
[1711] d]pyrimidin-4(3H)-one H*> / ft ft
[1712] 6 3.65 - 3.85 (m, 4H), 2.71 - 2.95 (m, 1 H), 1.80 - 2.1 1 (m, 2H), 1.28 - 1.42 (m, 3H), 1.10 - 1.26 (m, 6H).
[1713] 70 69 (S)-3-ethyl-8-((4- 409.15 1 H NMR (300 MHz, DMSO- isopropylphenyl)amino) [M+H]+d6) 6 (ppm): 8.40 (s, 1 H), -6-(3- 7.88 - 7.91 (m, 2H), 7.26 - Tin
[1714] methoxypyrrolidin-1 - A N 7.29 (m, 2H), 4.01 - 4.15 yl)pyrimido[5,4-zX--1' 6 (m, 3H), 3.61 - 3.95 (m, 4H), d]pyrimidin-4(3H)-one 3.27 (s, 3H), 2.71 - 2.99 (m,
[1715] 1 H), 1.90 - 2.22 (m, 2H),
[1716]
[1717] PATENT
[1718] ATTORNEY-DOCKET NO.: 51478-036WO4
[1719] 1.28 - 1.41 (m, 3H), 1.07 - 1.22 (m, 6H).
[1720] 71 70 6-(dimethylamino)-3- 353.15 1H NMR (400 MHz, DMSO- ethyl-8-((4- [M+H]+d6) 6 (ppm): 8.38 - 8.40 (m, isopropylphenyl)amino) 1. 1H), 7.81 - 7.84 (m, 2H), pyrimido[5,4- ' rj 7.26 - 7.28 (m, 2H), 4.02 - A A.
[1721] d]pyrimidin-4(3H)-one H A A 4.07 (m, 2H), 3.21 - 3.24 (s,
[1722] ;O 6H), 2.85 - 2.92 (m, 1H),
[1723] 1.29 - 1.32 (m, 3H), 1.20 - 1.22 (m, 6H).
[1724] 72 71 3-ethyl-6-((2- 383.10 1H NMR (400 MHz, DMSO- hydroxyethyl)(methyl)a [M+H]+d6) 6 (ppm): 8.40 - 8.44 (m, mino)-8-((4- 1H), 7.82 - 7.84 (m, 2H), isopropylphenyl)amino) 7.26 - 7.28 (m, 2H), 4.02 -ACiKK
[1725] pyrimido[5,4- 4.07 (m, 2H), 3.72 - 3.82 d]pyrimidin-4(3H)-one 6 (m, 4H), 3.23 - 3.25 (s, 3H),
[1726] 2.86 - 2.92 (m, 1H), 1.29 - 1.32 (m, 3H), 1.20 - 1.22 (m, 6H).
[1727] 73 72 3-ethyl-8-((4- 395.15 1H NMR (400 MHz, DMSO- isopropylphenyl)amino) [M+H]+d6) 6 (ppm): 8.47 (s, 1 H), -6-(3-methoxyazetidin- 7.80 - 7.82 (m, 2H), 7.30 - 1-yl)pyrimido[5,4- 7.33 (m, 2H), 4.41 - 4.60 d]pyrimidin-4(3H)-one (m, 2H), 4.26 - 4.39 (m, 1H), ft <. N z 4.02 - 4.16 (m, 4H), 3.29 (s,
[1728] 3H), 2.85 - 2.96 (m, 1H), 1.31 - 1.42 (m, 3H), 1.12 - 1.30 (m, 6H).
[1729] 74 73 1-(7-ethyl-4-((4- 436.15 1H NMR (400 MHz, DMSO- isopropylphenyl)amino) [M+H]+d6) 6 (ppm): 8.48 (s, 1 H), -8-oxo-7,8- 7.88 - 7.94 (m, 2H), 7.28 - dihydropyrimido[5,4- 7.33 (m, 2H), 4.20 - 4.53 d]pyrimidin-2-yl)-N, N- | C’N H A Y (m, 4H), 4.00 - 4.18 (m, 2H), dimethylazetidine-3- 0 3.80 - 3.93 (m, 1H), 2.82 - carboxamide 2.99 (m, 7H), 1.24 - 1.38 (m, 3H), 1.11 - 1.23 (m, 6H).
[1730] 75 74 (R)-8-((4-(3,3- R 443.15 1H NMR (300 MHz, DMSO- F V-C
[1731] difluorocyclobutyl)phen [M+H]+d6) 6 (ppm): 9.56 - 9.57 (m, yl)amino)-3-methyl-6- A 1H), 8.21 - 8.56 (m, 1H), (2- > 6 7.79 - 7.88 (m, 2H), 7.29 - i
[1732] methylmorpholino)pyri 7.36 (m, 2H), 4.40 - 4.52
[1733]
[1734] PATENT
[1735] ATTORNEY-DOCKET NO.: 51478-036WO4
[1736] mido[5,4-d]pyrimidin- (m, 2H), 3.90 - 3.93 (m, 1H), 4(3H)-one 3.45 - 3.49 (m, 5H), 3.38 - 3.41 (m, 1H), 2.93 - 3.17 (m, 3H), 2.50 - 2.77 (m, 3H), 1.18 - 1.23 (m, 3H).
[1737] 76 75 3-ethyl-6-(methyl(2,2,2- 447.05 1H NMR (300 MHz, DMSO- trifluoroethyl)amino)-8- [M+H]+d6) 6 (ppm): 10.12 (s, 1H), ((4- o 8.33 (s, 1H), 8.15 - 8.17 (m, (trifluoromethyl)phenyl) 2H), 7.69 - 7.72 (m, 2H), amino)pyrimido[5,4- 4.55 - 4.66 (m, 2H), 4.01 - d]pyrimidin-4(3H)-one 4.06 (m, 2H), 3.22 - 3.26 (m, 3H), 1.23 - 1.32 (m, 3H).
[1738] 77 76 (R)-3-methyl-6-(2- 435.05 1H NMR (300 MHz, DMSO- methylmorpholino)-8- [M+H]+d6) 6 (ppm): 8.24 (s, 1 H), ((4-(2,2,2- 7.88 - 7.90 (m, 2H), 7.34 - trifluoroethyl)phenyl)a 7.37 (m, 2H), 4.39 - 4.51 mino)pyrimido[5,4- (m, 2H), 3.90 - 3.94 (m, 1H), d]pyrimidin-4(3H)-one r 3.59 - 3.63 (m, 7H), 2.95 - 3.04 (m, 1H), 2.64 - 2.71 (m, 1H), 1.15 - 1.17 (m, 3H).
[1739] 78 77 8-((3- 393.15 1H NMR (300 MHz, DMSO- cyclopropylphenyl)ami [M+H]+d6) 6 (ppm): 8.29 (s, 1 H), no)-3-ethyl-6- 7.58 - 7.61 (m, 2H), 7.22 - morpholinopyrimido[5,4 7.29 (m, 1H), 6.88 - 6.91 -d]pyrimidin-4(3H)-one (m, 1H), 3.95 - 4.05 (m, 2H),
[1740] 3.72 - 3.85 (m, 8H), 1.85 - 2.01 (m, 1H), 1.22 - 1.40 (m, 3H), 0.90 - 1.05 (m, 2H), 0.61 - 0.71 (m, 2H).
[1741] 79 78 6-(diethylamino)-8-((4- 383.15 1H NMR (300 MHz, DMSO)
[1742] (1- [M+H]+69.76 (s, 1H), 8.25 (s, 1H), fluorocyclopropyl)phen Vi 7.97 - 8.00 (m, 2H), 7.28 - yl)amino)-3- A 7.31 (m, 2H), 3.62 - 3.66 methylpyrimido[5,4- (m, 4H), 3.51 (s, 3H), 1.38 - d]pyrimidin-4(3H)-one 1.47 (m, 2H), 1.11 - 1.19 (m, 8H).
[1743] 80 79 3-ethyl-6- 393.05 1H NMR (400 MHz, DMSO)
[1744] (ethyl(methyl)amino)-8- [M+H]+6: 9.87 (s, 1H), 8.22 - 8.24 ((4- (m, 3H), 7.70 - 7.73 (m, 2H), (trifluoromethyl)phenyl) 4.02 - 4.04 (m, 2H), 3.67 -
[1745]
[1746] PATENT
[1747] ATTORNEY-DOCKET NO.: 51478-036WO4
[1748] amino)pyrimido[5,4- 3.72 (m, 2H), 3.15 (s, 3H), d]pyrimidin-4(3H)-one 1.13 - 1.17 (m, 6H).
[1749] 81 80 (S)-3-methyl-6-(2- 375.00 1H NMR (400 MHz, DMSO- (trifluoromethyl)morpho [M+H]+ d6) 6 (ppm): 10.08 (s, 1H), lino)-8-((4- 8.26 (s, 1H), 8.10 - 8.12 (m, (trifluoromethyl)phenyl) 2H), 7.67 - 7.69 (m, 2H), amino)pyrimido[5,4- 4.67 - 4.70 (m, 1H), 4.42 - d]pyrimidin-4(3H)-one 6 r. A >'V / 6 '' 4.46 (m, 1H), 4.29 - 4.30
[1750] X CF3(m, 1H), 4.06 - 4.09 (m, 1H),
[1751] 3.67 - 3.69 (m, 1H), 3.50 (s, 3H), 3.09 - 3.20 (m, 2H). 82 81 (R)-3-methyl-6-(2- 375.00 1H NMR (300 MHz, DMSO- (trifluoromethyl)morpho [M+H]+d6) 6 (ppm): 8.27 (s, 1 H), lino)-8-((4- 8.10 - 8.13 (m, 2H), 7.68 - (trifluoromethyl)phenyl) 7.71 (m, 2H), 4.67 - 4.72 amino)pyrimido[5,4- (m, 1H), 4.43 - 4.47 (m, 1H),
[1752] N' H' y '
[1753] d]pyrimidin-4(3H)-one O 4.27 - 4.32 (m, 1H), 4.07 - 4.10 (m, 1H), 3.64 - 3.71 (m, 1H), 3.45 - 3.51 (m, 3H), 3.10 - 3.24 (m, 2H).
[1754] 83 82 8-((4-(3,3- 447.15 1H NMR (400 MHz, DMSO) difluorocyclobutyl)-3- E [M+H]+6: 9.82 (s, 1H), 8.24 (s, 1H), fluorophenyl)amino)-3- 7.76 - 7.88 (m, 2H), 7.34 - methyl-6- 7.38 (m, 1H), 3.65 - 3.79
[1755] A
[1756] morpholinopyrimido[5,4 (m, 8H), 3.40 - 3.80 (m, 4H),
[1757] 6
[1758] -d]pyrimidin-4(3H)-one 2.94 - 3.05 (m, 2H), 2.71 - 2.86 (m, 2H).
[1759]
[1760] Example 84: 3-isopropyl-8-((4-isopropylphenyl)amino)-6-(2-methylpyridin-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 2)
[1761] Step 1. Synthesis of 3-isopropyl-8-methoxy-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1762] OH
[1763] r^Y®'
[1764] N,.;JOH
[1765] V Pd(dppf)Ci2, K3PO4, dioxane / H2O0crV'v’V'. * J X il -Cl Nn f 90’C, 2 hrsN]T Y
[1766] 0N 6!
[1767]
[1768] A solution of 6-chloro-3-isopropyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (200 mg, 0.785 mmol, 1 equiv), Pd(dppf)Cl2 (57.46 mg, 0.079 mmol, 0.1 equiv), K3PO4 (500.09 mg, 2.355 mmol, 3 equiv)PATENT
[1769] ATTORNEY-DOCKET NO.: 51478-036WO4
[1770] and (2-methylpyridin-4-yl) boronic acid (118.31 mg, 0.864 mmol, 1.1 equiv) in Dioxane (3 mL) / H2O (0.6 mL) was stirred at 90°C for 2h under nitrogen atmosphere. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The resulting mixture was extracted with EtOAc (3 x 150mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 3-isopropyl-8-methoxy-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (150 mg, 61.35% yield, 92% purity) as a white solid. MS: MS (ESI) calcd. for C16H17N5O2: 311.14, Found: 312.15 [M+H]+.
[1771] Step 2. Synthesis of 8-hydroxy-3-isopropyl-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin- 4(3H)-one
[1772] OH
[1773] LiCI, TsOH, DMF 120’C, 1 hr
[1774]
[1775] A solution of 3-isopropyl-8-methoxy-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (120 mg, 0.385 mmol, 1 equiv), LiCI (81.69 mg, 1.925 mmol, 5 equiv) and TSOH (331.85 mg, 1.925 mmol, 5 equiv) in DMF (3 mL) was stirred at 120°C for 1h. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in 8-hydroxy-3-isopropyl-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 87.26% yield, 91% purity) as a white solid. MS: MS (ESI) calcd. for C15H15N5O2: 297.12, Found: 298.15 [M+H]+.
[1776] Step 3. Synthesis of 8-chloro-3-isopropyl-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1777]
[1778] A solution of 8-hydroxy-3-isopropyl-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.336 mmol, 1 equiv) in phosphoryl trichloride (0.5 mL) was stirred at 80°C for 1 h. The resulting mixture was concentrated under vacuum. The mixture was neutralized to pH 7 with saturated NaHCCh (aq.). The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 5% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in 8-chloro-3-isopropyl-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 94.16% yield, 92% purity) as a white solid. MS: MS (ESI) calcd. for C15H14CIN5O: 315.09, Found: 316.10 [M+H]+.PATENT
[1779] ATTORNEY-DOCKET NO.: 51478-036WO4
[1780] Step 4. Synthesis of 3-isopropyl-8-((4-isopropylphenyl)amino)-6-(2-methylpyridin-4-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1781]
[1782] A solution of 8-chloro-3-isopropyl-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (80 mg, 0.253 mmol, 1 equiv), para-isopropylaniline (37.68 mg, 0.278 mmol, 1.1 equiv) and DIEA (98.24 mg, 0.759 mmol, 3 equiv) in DMSO (2 mL) was stirred at 100°C for 1h. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH4HCO3), 2% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in 3-isopropyl-8-((4-isopropylphenyl) amino)-6-(2-methylpyridin-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (31.8 mg, 30.28% yield, 99.7% purity) as a white solid. LC / MS: MS (ESI) calcd. for C24H26N6O: 414.22. Found: 415.20 [M+H]+ 1H NMR (300 MHz, DMSO-d₆) 6 (ppm): 9.89 (s, 1H), 8.58 - 8.64 (m, 2H), 8.10 (s, 1H), 8.01 - 8.03 (m, 1H),7.93 - 7.96 (m, 2H), 7.32 - 7.35 (m, 2H), 4.99 - 5.04 (m, 1H), 2.88 - 2.97 (m, 1H), 2.60 (s, 3H), 1.48 - 1.51 (m, 6H), 1.21 - 1.26 (m, 6H).
[1783] Example 85: (R)-8-((4-(3,3-difluorocyclobutyl)phenyl)amino)-3-ethyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 32)
[1784] Step 1. Synthesis of 1-bromo-4-(3,3-difluorocyclobutyl) benzene
[1785]
[1786] To a solution of 3-(4-bromophenyl) cyclobutan-1-one (600 mg, 2.666 mmol, 1 equiv) in 5 mL of DCM (5 mL) and cooled to -70 °C. DAST (1074.19 mg, 6.665 mmol, 2.5 equiv) was added drop wise and the reaction mixture was stirred at room temperature for 24 h. After completion of reaction monitored by LCMS. The reaction mixture was quenched with NH4HCO3 (50 mL), extracted with DCM (3 x 50 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 1 -bromo-4-(3,3-difluorocyclobutyl) benzene (450 mg, 68.32% yield, 95% purity) as a white solid. MS: MS (ESI) calcd. for C7H6BrCIN2O3: 223.98, Found: 224.05 [M+H]+.PATENT
[1787] ATTORNEY-DOCKET NO.: 51478-036WO4
[1788] Step 2. Synthesis of tert-butyl N-[4-(3,3-difluorocyclobutyl) phenyl] carbamate
[1789] , F- ')
[1790] NHjBoc, BrettPhos Pd G3, CSJCOJ, dioxane \_.
[1791] " X I. A y
[1792]
[1793] ^An 80*C, 24 hr N O '•
[1794] ' 8r H To a solution of 1-bromo-4-(3,3-difluorocyclobutyl) benzene (400 mg, 1.619 mmol, 1 equiv) in 15 mL of dioxane (15 mL) were added NH2B0C (751.13 mg, 2.429 mmol, 1.5 equiv), BrettPhos Pd G3 (293.51 mg, 0.324 mmol, 0.2 equiv) and Cs₂CO₃ (1054.93 mg, 3.238 mmol, 2 equiv) at room temperature. The resulting mixture was stirred at 80°C for 2 h. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (50 mL). The aqueous layer was extracted with ethyl acetate (3 x 150 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN in Water (0.05% NH4HCO3), 5% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in tert-butyl N-[4-(3,3-difluorocyclobutyl) phenyl] carbamate (250 mg, 54.51% yield, 95% purity) as a yellow oil. MS: MS (ESI) calcd. for C15H19F2NO2: 283.14, Found: 282.00 [M-H]+.
[1795] Step 3. Synthesis of 4-(3,3-difluorocyclobutyl) aniline
[1796]
[1797] H
[1798] A solution of tert-butyl (4-(3,3-difluorocyclobutyl) phenyl) carbamate (100 mg, 2.022 mmol, 1 equiv) in 8 mL of DCM (6 mL) and TFA (2 mL) was stirred at 25°C for 2 h. After completion of reaction, the reaction was quenched with sat. NH4CI (aq.) at 0°C. The resulting mixture was extracted with EA (3 x 100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford 4-(3,3-difluorocyclobutyl) aniline (75 mg, 29.00% yield, 95% purity) as a white solid. MS: MS (ESI) calcd. for C10H11F2N: 183.09, Found: 184.30 [M+H]+.
[1799] Step 4. Synthesis of (R)-8-((4-(3,3-difluorocyclobutyl)phenyl)amino)-3-ethyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1800]
[1801] To a solution of 8-chloro-3-ethyl-6-[(2R)-2-methylmorpholin-4-yl] pyrimido[5,4-d] [1,3] diazin-4-one (60 mg, 0.194 mmol, 1 equiv) and 4-(3,3-difluorocyclobutyl) aniline (53.23 mg, 0.291 mmol, 1.5 equiv) in 1.5 mL of DMSO (1.5 mL) was added DIEA (50.07 mg, 0.388 mmol, 2 equiv). The resulting mixture wasPATENT
[1802] ATTORNEY-DOCKET NO.: 51478-036WO4
[1803] stirred at 100°C for2h. LCMS showed the reaction was completed. The mixture solution was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN in Water (0.1% TFA), 5% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-8-((4-(3,3-difluorocyclobutyl) phenyl) amino)-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (12.3 mg, 13.91% yield, 99.6% purity) as a white solid. LC / MS: MS (ESI) calcd. for C23H26F2N6O2: 456.21. Found: 457.25 [M+H]+ 1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 9.57 - 9.60 (m, 1H), 8.11 - 8.38 (m, 1H), 7.72 - 8.01 (m, 2H), 7.17 - 7.48 (m, 2H), 4.28 - 4.68 (m, 2H), 3.99 - 4.19 (m, 2H), 3.79 - 3.99 (m, 1H), 3.38 - 3.59 (m, 3H), 2.87 - 3.19 (m, 3H), 2.59 - 2.87 (m, 3H), 1.21 - 1.39 (m, 3H), 1.09 - 1.21 (m, 3H).
[1804] Example 86: (R)-3-methyl-6-(2-methylmorpholino)-8-((4-(trifluoromethyl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 87)
[1805] Step 1. Synthesis of 6-chloro-3-methyl-8-((4-(trifluoromethyl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1806] G 'NH- DIEA, DMSO
[1807] ZN. J J
[1808] ' ** Y 50’0, 2 hr cr 'N'
[1809]
[1810] c 6
[1811] To a solution of 6,8-dichloro-3-methylpyrimido[5,4-d] [1,3] diazin-4-one (50 mg, 0.216 mmol, 1 equiv) and P-trifluoromethylaniline (34.87 mg, 0.216 mmol, 1 equiv) in DMSO (1 mL) was added DIEA (83.91 mg, 0.648 mmol, 3 equiv) was stirred at 50°C for 2 h. Desired product could be detected by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in 6-chloro-3-methyl-8-((4-(trifluoromethyl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (30.2 mg, 39.23% yield, 95% purity) as an off-white solid. MS: MS (ESI) calcd. for C14H9CIF3N5O: 355.04, Found: 356.00 [M+H]+.
[1812] Step 2. Synthesis of (R)-3-methyl-6-(2-methylmorpholino)-8-((4-(trifluoromethyl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1813] NH p
[1814] T > r ||
[1815] I DIEA, DMSO
[1816] A A N
[1817] we e. 1 hr f- M' fcr Y
[1818] 0, J O
[1819]
[1820] To a solution of 6-chloro-3-methyl-8-{[4-(trifluoromethyl) phenyl] amino} pyrimido[5,4-d] [1,3] diazin-4-one (30 mg, 0.084 mmol, 1 equiv) and (2R)-2-methylmorpholine (25.59 mg, 0.252 mmol, 3 equiv) in DMSO (1 mL) was added DIEA (32.70 mg, 0.252 mmol, 3 equiv) was stirred at 100°C for 1h. Desired product could be detected by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% toPATENT
[1821] ATTORNEY-DOCKET NO.: 51478-036WO4
[1822] 50% gradient in 10 min; detector, UV 254 nm. This resulted in (R)-3-methyl-6-(2-methylmorpholino)-8-((4-(trifluoromethyl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (12.6 mg, 42.64% yield, 99.5% purity) as a trifluoroacetate salt and as an off-white solid. MS: MS (ESI) calcd. for C19H19F3N6O2: 420.15, Found: 421.10 [M+H]+ 1H NMR (400 MHz, DMSO) 6 9.98 (s, 1H), 8.25 (s, 1H), 8.14 - 8.16 (m, 2H), 7.72 -7.74 (m, 2H), 4.41 - 4.51 (m, 2H), 3.91 - 3.94 (m, 1H), 3.40 - 3.58 (m, 5H), 2.97 - 3.04 (m, 1H), 2.65 - 2.70 (m, 1H), 1.19 - 1.23 (m, 3H).
[1823] Example 87: 3-ethyl-8-((4-isopropylphenyl)amino)-6-(methyl(oxetan-3-yl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 88)
[1824] Step 1. Synthesis of 6-chloro-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1825] ■XO V
[1826] G8
[1827] nOKA, DM3C
[1828] CS '' " N100-C(
[1829]
[1830] o a
[1831] A solution of 6,8-dichloro-3-ethylpyrimido[5,4-d] pyrimidin-4(3H)-one (430 mg, 1.755 mmol, 1 equiv), 4-isopropylaniline (237.3 mg, 1.755 mmol, 1 equiv) and DIEA (680.4 mg, 5.265 mmol, 3 equiv) in DMSO (1 mL) was stirred at 100°C for 1 h. Desired product could be detected by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, water (0.05% TFA) in ACN, 0% to 60% gradient in 15 min; detector, UV 254 nm. This resulted in 6-chloro-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (500 mg, 82.88% yield, 80% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C17H18CIN5O: 343.12, Found: 344.00[M+H]+.
[1832] Step 2. Synthesis of 3-ethyl-8-((4-isopropylphenyl) amino)-6-(methyl(oxetan-3-yl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1833]
[1834] To a solution of 6-chloro-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (70 mg, 0.204 mmol, 1 equiv), N-methyloxetan-3-amine (70.9 mg, 0.816 mmol, 4 equiv) in DMSO (2 mL) was added DIEA (78.9 mg, 0.612 mmol, 3 equiv) was stirred at 100°C for 2 h. Desired product could be detected by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, water (0.05% TFA) in ACN, 10% to 50% gradient in 10 min; detector, UV 254, 220 nm. This resulted in 3-ethyl-8-((4-isopropylphenyl) amino)-6-(methyl(oxetan-3-yl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (75.7 mg, 94.26% yield, 97.5% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C21H26N6O2: 394.21, Found: 395.15[M+H]+.1H NMR (400 MHz, DMSO-de) 6 (ppm): 11.08 - 11.13 (m, 1 H), 8.55 - 8.61 (m, 1 H), 7.75 - 7.85 (m, 2H), 7.30 - 7.39 (m, 2H),PATENT
[1835] ATTORNEY-DOCKET NO.: 51478-036WO4
[1836] 4.73 - 4.85 (m, 1 H), 4.60 - 4.70 (m, 1 H), 4.18 - 4.26 (m, 1 H), 4.05 - 4.15 (m, 2H), 3.80 - 3.88 (m, 1 H), 3.55 - 3.63 (m, 1H), 3.02 - 3.11 (m, 3H), 2.90 - 3.00(m, 1H), 1.30 - 1.40 (m, 3H), 1.19 - 1.28 (m, 6H).
[1837] Example 88: 3-ethyl-8-((4-isopropylphenyl)amino)-6-(oxetan-3-ylamino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 89)
[1838] Synthesis of 3-ethyl-8-((4-isopropylphenyl)amino)-6-(oxetan-3-ylamino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1839] i
[1840] A DIE*. WOO
[1841] t 1.... * IJ 1
[1842] «rc.2hf 'N' V
[1843] O A 6
[1844]
[1845] 'o''
[1846] To a solution 6-chloro-3-ethyl-8-[(4-isopropylphenyl) amino] pyrimido[5,4-d] [1,3] diazin-4-one (70 mg, 0.204 mmol, 1 equiv) and oxetan-3-amine (29.7 mg, 0.408 mmol, 2 equiv) in DMSO (1.5 mL) was added DIEA (78.9 mg, 0.612 mmol, 3 equiv) at room temperature. The reaction mixture was stirred 80 °C for a period of 2 h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 30 min; detector, UV 254 nm. To afford 3-ethyl-8-[(4-isopropylphenyl) amino]-6-(oxetan-3-ylamino) pyrimido[5,4-d] [1,3] diazin-4-one (51.2 mg, 50.86% yield, 97.3% purity) as a trifluoroacetate salt and as a white solid. LC / MS: MS (ESI) calcd. for C20H24N6O2: 380.19 m / z, found: 381,15[M+H]+.1H NMR (400 MHz, DMSO-de) 6 (ppm): 8.60 - 8.70 (m, 1H), 7.60 - 8.00 (m, 2H), 7.20 - 7.40 (m, 2H), 4.60 - 4.90 (m, 2H), 4.20 - 4.30 (m, 1 H), 3.95 - 4.20 (m, 2H), 3.60 - 3.70 (m, 2H), 2.80 - 3.05 (m, 1 H), 1.30 - 1.50 (m, 3H), 1.00 - 1.30 (m, 6H).
[1847] Example 89: 3-ethyl-8-((4-isopropylphenyl)amino)-6-(oxetan-3-yloxy)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 90)
[1848] Synthesis of 3-ethyl-8-((4-isopropylphenyl) amino)-6-(oxetan-3-yloxy) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1849] OH O
[1850] S-BuOK, THF
[1851]
[1852] To a solution of 6-chloro-3-ethyl-8-((4-isopropylphenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.291 mmol) in THF (10 mL) were added t-BuOK (163.2 mg, 1.455 mmol) and oxetan-3-ol (861.8 mg, 11.640 mmol) at 20 °C. The mixture was replaced with N2 for 3 times. The resulting mixture was stirred at 80 °C for 3 h. The reaction was monitored by LCMS. After completion of reaction, the mixture was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, water (0.05% NH4HCO3) in ACN, 30% to 80% gradient in 20 min; detector, UV 254 nm. After lyophilization, it provided 3-ethyl-8-((4-isopropylphenyl) amino)-6-(oxetan-3-yloxy)PATENT
[1853] ATTORNEY-DOCKET NO.: 51478-036WO4
[1854] pyrimido[5,4-d] pyrimidin-4(3H)-one (52.4 mg, 47.23% yield, 98.3% purity) as a white solid. LC / MS: MS (ESI) calcd for C20H23N5O3: 381.18. Found: 382.05[M+H]+.1H NMR (400 MHz, DMSO-d6) 6: 9.91 (s, 1 H), 8.43 - 8.44 (m, 1 H), 7.72 - 7.74 (m, 2H), 7.26 - 7.28 (m, 2H), 5.53 - 5.58 (m, 1 H), 4.83 - 4.87 (m, 2H), 4.56 - 4.59 (m, 2H), 4.00 - 4.06 (m, 2H), 2.86 - 2.93 (m, 1 H), 1.27 - 1.31 (m, 3H), 1.19 - 1.22 (m, 6H).
[1855] Using the approach to synthesize Compound 29, the compounds of Table 6 were synthesized from morpholine and the corresponding aniline.
[1856] Table 6: Experimental Data for Compounds
[1857] Ex Cmpd
[1858] Name Structure MS 1H NMR
[1859] # #
[1860] 90 91 8-((3-chlorophenyl)- 401.10 1 H NMR (300 MHz, DMSO- amino)-3-ethyl-2- [M+H]+d6) 6: 8.01 - 8.08 (m, 1 H), 7.83 methyl-6-morpholino- sZV' NH - 7.85 (m, 1 H), 7.40 - 7.46 (m, pyrimido[5,4- 1 H), 7.18 - 7.21 (m, 1 H), 4.00 -.3 A f.t
[1861] d]pyrimidin-4(3H)-one f N N" 'if 4.08 (m, 2H), 3.71 - 3.74 (m,
[1862] 6
[1863] 8H), 2.75 (s, 3H), 1.24 - 1.28 (m, 3H).
[1864] 91 92 8-((4- 407.15 1 H NMR (300 MHz, DMSO-d6) cyclopropylphenyl)am [M+H]+6: 7.69 - 7.70 (m, 2H), 7.1 1 - ino)-3-ethyl-2-methyl- 7.14 (m, 2H), 4.07 - 4.14 (m, 'NH
[1865] 6-. / LAX 2H), 3.72 - 3.74 (m, 8H), 2.68 morpholinopyrimido[5..-"v A A, N.
[1866] rnr (s, 3H), 1.90 - 1.95 (m, 1 H),4-d]pyrimidin-4(3H)- 1.25 - 1.29 (m, 3H), 0.95 - 0.99 one (m, 2H), 0.69 - 0.70 (m, 2H). 92 93 8-((3- 407.15 1 H NMR (400 MHz, DMSO-d6) cyclopropylphenyl)am [M+H]+6 (ppm):9.21 - 9.28 (m, 1 H), ino)-3-ethyl-2-methyl- 7.55 - 7.67 (m, 2H), 7.18 - 7.27 6- (m, 1 H), 6.83 - 6.91 (m, 1 H), morpholinopyrimido[5 4.01 - 4.13 (m, 2H), 3.70 - 3.89 N X v,. x-i xT.
[1867] ,4-d]pyrimidin-4(3H)- f N N - (m, 4H), 3.65 - 3.70 (m, 4H), one C^ J Q 2.60 - 2.79 (s, 3H), 1.88 - 1.98(m, 1 H), 1.11 - 1.40 (m, 3H), 0.85 - 1.10 (m, 2H), 0.59 - 0.80 (m, 2H).
[1868]
[1869] PATENT
[1870] ATTORNEY-DOCKET NO.: 51478-036WO4
[1871] Example 93: (R)-3-ethyl-8-((4-isopropylphenyl)amino)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 94)
[1872] Step 1. Synthesis of 4-bromo-1-(difluoromethyl)-2 -fluorobenzene
[1873] 9F
[1874] DAST, DCMF
[1875]
[1876] 0*C - FL 18 hrsF'8r
[1877] To a stirred solution of 4-bromo-2-fluorobenzaldehyde (500 mg, 2.463 mmol, 1 equiv) in anhydrous DCM (10 mL) was added DAST (794.00 mg, 4.926 mmol, 2 equiv) at 0 °C. The reaction mixture was stirred at 25 °C for a period of 16 h. Desired product could be detected by GCMS. After completion of reaction, the reaction mixture was quenched by addition of saturated NaHCOs (25 mL) at 0 °C. The aqueous layer was extracted with ethyl acetate (3 x 100 mL). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by silica gel column chromatography, eluted with PE to afford 4-bromo-1-(difluoromethyl)-2-fluorobenzene (370 mg, 66.77% yield, 93% purity) as a yellow oil. GC / MS: MS (ESI) calcd. for C7H5BrF2: 223.94, Found: 223.90.
[1878] Step 2. Synthesis of tert-butyl (4-(difluoromethyl)-3-fluorophenyl) carbamate
[1879] F F
[1880] 1 BretPbos Pd Cs^CO^ Sow
[1881] i >........................................................................ || j
[1882]
[1883] F' Br S0'C,ownl$tf: Fh <10
[1884] To a stirred solution of 4-bromo-1-(difluoromethyl)-2-fluorobenzene (200 mg, 0.889 mmol, 1 equiv) in anhydrous dioxane (5 mL) were added NH2B0C (156.19 mg, 1.333 mmol, 1.5 equiv) BrettPhos Pd G3 (120.86 mg, 0.133 mmol, 0.15 equiv) and Cs₂CO₃ (579.21 mg, 1.778 mmol, 2 equiv) at 25 °C. The reaction mixture was stirred at 80 °C for a period of 16 h. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL). The aqueous layer was extracted with ethyl acetate (3 x25 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by silica gel column chromatography, eluted with PE I EA (5 / 1) to afford tert-butyl (4-(difluoromethyl)-3-fluorophenyl) carbamate (154 mg, 66.32 % yield, 92% purity) as a white solid. LC / MS: MS (ESI) calcd. for C12H14F3NO2: 261.10, Found: 260.15 [M-H]+
[1885] Step 3. Synthesis of 4-(difluoromethyl)-3-fluoroaniline
[1886] j 1.
[1887] Q | HCIftHGxane ’^-' ^x
[1888]
[1889] H '
[1890] To a stirred solution of tert-butyl (4-(difluoromethyl)-3-fluorophenyl) carbamate (140 mg, 0.536 mmol, 1 equiv) in DCM (2 mL) was added HCI in 1,4-dioxane (4.0 M) (2 mL) at 0 °C. The resulting mixture was stirred at 25 °C for 1 h. The reaction was monitored by LCMS. After completion of reaction, the resulting mixture was concentrated under reduced pressure to afford 4-(difluoromethyl)-3-fluoroanilinePATENT
[1891] ATTORNEY-DOCKET NO.: 51478-036WO4
[1892] (60 mg, 69.49% yield, 93% purity) as a white solid. LC / MS: MS (ESI) calcd. for C7H6F2N: 161.05, Found: 162.10 [M+H]+
[1893] Step 4. Synthesis of (R)-8-((4-(difluoromethyl)-3-fluorophenyl)amino)-3-ethyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1894] Ci F 'vN H;. P
[1895] KaCQa, DMSO f N N jf WC, 16ftr < *hT 'N' y ”'i o J o16 J o '
[1896]
[1897] i T
[1898] To a stirred solution of (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (80 mg, 0.258 mmol, 1 equiv) in anhydrous DMSO (2 mL) were added 4-(difluoromethyl)-3-fluoroaniline (49.94 mg, 0.310 mmol, 1.2 equiv) and K2CO3 (71.39 mg, 0.516 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 70 °C for a period of 16 h. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL). The aqueous layer was extracted with ethyl acetate (3 x25 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford (R)-8-((4-(difluoromethyl)-3-fluorophenyl)amino)-3-ethyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one;trifluoroacetic acid salt (39.4 mg, 27.82% yield, 99.9% purity) as a light yellow solid. LC / MS: MS (ESI) calcd. for C20H21F3N6O2: 434.42, Found: 435.10.1H NMR (300 MHz, DMSO-d6) 6 (ppm): 8.26 - 8.28 (m, 1H), 7.89 - 7.98 (m, 1H), 7.85 - 7.87 (m, 1H), 7.59 -7.62 (m, 1H), 6.96 - 7.33 (m, 1H), 4.38 - 4.50 (m, 2H), 3.91 - 4.05 (m, 3H), 3.48 - 3.51 (m, 2H), 2.99 - 3.09 (m, 1 H), 2.67 - 2.75 (m, 1 H), 1.29 - 1.31 (m, 3H), 1.26 - 1.28 (m, 3H).
[1899] Example 94: (R)-8-((4-(difluoromethyl)-2-fluorophenyl)amino)-3-ethyl-6-(2-methylmorpholino)-pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 95)
[1900]
[1901] Using the same approach as for compound 91, starting from 4-bromo-3-fluorobenzaldehyde, (R)-8-((4-(difluoromethyl)-2-fluorophenyl) amino)-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one was isolated as a light yellow solid. LC / MS: MS (ESI) calcd. for C20H21F3N6O2: 434.42, Found: 435.10.1H-NMR (300 MHz, DMSO-d6) 6 (ppm): 8.19 - 8.27 (m, 2H), 7.48 - 7.58 (m, 2H), 6.84 - 7.21 (m, 1H), 4.33 - 4.45 (m, 2H), 3.87 - 4.04 (m, 2H), 3.69 - 3.70 (m, 1H), 3.47 - 3.70 (m, 2H), 2.93 - 3.03 (m, 1H), 2.52 - 2.69 (m, 1 H), 1.29 - 1.32 (m, 3H), 1.26 - 1.28 (m, 3H).PATENT
[1902] ATTORNEY-DOCKET NO.: 51478-036WO4
[1903] Example 95: 3-ethyl-8-((4-((1s,3S)-3-fluorocyclobutyl)phenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-ethyl-8-((4-((1 r,3R)-3-fluorocyclobutyl)phenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 96 and 97)
[1904] Step 1. Synthesis of tert-butyl (4-(3-fluorocyclobutyl) phenyl) carbamate
[1905] '"■'" 'OH
[1906] F.„,
[1907] BrWHC, pyrMrte. g& OMe, rt, 15min V-A. Q
[1908] cqt&iucWte, jjfiWSmricte. BuOW H
[1909]
[1910] DMA Wue LEDs, rts16hrs
[1911] To a solution of 3-fluorocyclobutan-1-ol (253.27 mg, 2.810 mmol, 2.55 equiv) in MTBE (43.6 mL) was added 5, 7-di-tert-butyl-3-phenylbenzo[d]oxazol-3-ium tetrafluoroborate (1045.70 mg, 2.645 mmol, 2.4 equiv) in portions at 0°C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 10min under nitrogen atmosphere. To this suspension, a solution of pyridine (209.28 mg, 2.645 mmol, 2.4 equiv) in MTBE (43.6 mL) was added dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 15 min under nitrogen atmosphere. Another dram vial was charged with tert-butyl (4-bromophenyl) carbamate (300 mg, 1.102 mmol, 1 equiv), NiBr2(dtbbpy) (26.84 mg, 0.055 mmol, 0.05 equiv), [lr(dtbbpy)(ppy)2]PFe (15.11 mg, 0.017 mmol, 0.015 equiv), Quinuclidine (214.50 mg, 1.929 mmol, 1.75 equiv) and Phthalimide (35.68 mg, 0.242 mmol, 0.22 equiv). DMAc (13.5 mL) was added and the reaction mixture was sparged with nitrogen atmosphere. The suspension from the first vial was transferred to a syringe. Then a syringe filter and new needle were installed on the syringe, and 1 mL of the solution was injected through the syringe filter into the DMAc solution. The resulting reaction mixture was sparged with nitrogen for 15 min. The vial was irradiated with 2 blue LEDs Photoreaction at room temperature for 16 hours. LCMS showed that the desired product was formed. The reaction mixture was quenched by addition of saturated NH4CI (20 mL). The aqueous layer was extracted with EtOAc (3 x 150 mL). The combined organic phase was washed with brine (70 mL), dried over anhydrous Na2SC>4. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (5:1) to afford tert-butyl (4-(3-fluorocyclobutyl) phenyl) carbamate (250 mg, 85.47% yield, 94% purity) as a white solid. LC / MS: MS (ESI) calcd for C15H20FNO2: 265.15. Found: 266.10[M+H]+.
[1912] Step 2. Synthesis of 4-(3-fluorocyclobutyl) aniline
[1913] F^ F,v,
[1914] TA Q HCiZdiasane V '■*'I. '-x
[1915] LL M A O' JA '' ft, 1 hr it, A NH2
[1916]
[1917] H
[1918] A solution of tert-butyl (4-(3-fluorocyclobutyl) phenyl) carbamate (250 mg, 0.942 mmol, 1 equiv) in HCI in 1,4-dioxane (4.0 M) (5 mL) was stirred at room temperature for 1 h under nitrogen atmosphere. LCMS showed that the desired product was formed. The resulting mixture was concentrated underPATENT
[1919] ATTORNEY-DOCKET NO.: 51478-036WO4
[1920] reduced pressure. This resulted in 4-(3-fluorocyclobutyl) aniline (150 mg, crude product) as a yellow solid. LC / MS: MS (ESI) calcd forCioHi2FN: 165.10. Found: 166.10[M+H]+.
[1921] Step 3. Synthesis of (R)-3-ethyl-8-((4-(3-fluorocyclobutyl) phenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1922] R-. R
[1923] Cl - NNDIEA. DMSON
[1924] J. J
[1925]
[1926] 1 I
[1927] To a solution of (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (150 mg, 0.484 mmol, 1 equiv) and 4-(3-fluorocyclobutyl) aniline (96.01 mg, 0.581 mmol, 1.2 equiv) in DMSO (3 mL) was added DIEA (125.18 mg, 0.968 mmol, 2 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 70°C for 5 h under nitrogen atmosphere. After completion of reaction, the reaction mixture was quenched by addition of saturated NF CI (2 mL) and extracted with ethyl acetate (30 mL x 3). The combined extract was washed with water, brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in (R)-3-ethyl-8-((4-(3-fluorocyclobutyl) phenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (150 mg, 70.64% yield, 96% purity) as a yellow solid. LC / MS: MS (ESI) calcd for C23H27FN6O2: 438.22. Found: 439.20[M+H]+
[1928] Step 4. Separation of 3-ethyl-8-((4-((1s,3S)-3-fluorocyclobutyl)phenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-ethyl-8-((4-((1 r,3R)-3-fluorocyclobutyl)phenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1929]
[1930] The product (150 mg) was purified by silica gel Column: CHIRALPAK AD-H, 5 pm, 250 mm x 20 mm; Mobile Phase A: Hex (with 0.1% TFA), Mobile Phase B: ETCH; Flow rate: 20 mL / min; Gradient (B%)30% B; Wave Length: 220 / 254 nm; Sample Solvent: EtOH--HPLC; Injection Volume: 0.5 mL;
[1931] Number Of Runs: 14; Single injection run time(min): 29 to afford 3-ethyl-8-((4-((1s,3S)-3fluorocyclobutyl)phenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (5.4 mg, 2.86% yield, 99.8% purity) as a yellow solid. Stereochemistry arbitrarily assigned.
[1932] RT1(min): 22.24;
[1933] LC / MS: MS (ESI) calcd for C23H27FN6O2: 438.22. Found: 439.15[M+H]+PATENT
[1934] ATTORNEY-DOCKET NO.: 51478-036WO4
[1935] 1H NMR (300 MHz, DMSO+D2O) 6: 8.29 (s, 1H), 7.76 - 7.86 (m, 2H), 7.24 - 7.33 (m, 2H), 5.20 - 5.40 (m, 1H), 4.40 - 4.48 (m, 2H), 3.87 - 4.08 (m, 2H), 3.76 - 3.87 (m, 1H), 3.60 - 3.70 (m, 2H), 3.41 - 3.60 (m, 1 H), 2.98 - 3.07 (m, 1 H), 2.61 - 2.76 (m, 3H), 2.52 - 2.61 (m, 2H), 1.27 - 1.31 (m, 3H), 1.24 - 1.27 (m, 3H).
[1936] And also to obtain the assumed 3-ethyl-8-((4-((1r,3R)-3-fluorocyclobutyl)phenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (45.2 mg, 23.91% yield, 98.7% purity) as an yellow solid. Stereochemistry arbitrarily assigned.
[1937] RT2(min): 26.48
[1938] LC / MS: MS (ESI) calcd for C23H27FN6O2: 438.22. Found: 439.15[M+H]+
[1939] 1H NMR (300 MHz, DMSO+D2O) 6: 8.35 (s, 1H), 7.77 - 7.81 (m, 2H), 7.29 - 7.33 (m, 2H), 4.94 - 5.18 (m, 1H), 4.37 - 4.48 (m, 2H), 3.91 - 4.08 (m, 3H), 3.41 - 3.60 (m, 2H), 2.98 - 3.20 (m, 2H), 2.61 - 2.77 (m, 3H), 2.12 - 2.31 (m, 2H), 1.27 - 1.31 (m, 3H), 1.24 - 1.27 (m, 3H).
[1940] Example 96: 3-methyl-6-((R)-2-methylmorpholino)-8-((4-((R)-1,1,1 -trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-methyl-6-((R)-2-methylmorpholino)-8-((4-((S)-1,1,1 -trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 98 and 99)
[1941] Step 1. Synthesis of 4-(3,3,3-trifluoroprop-1-en-2-yl)aniline
[1942] J. 1 fi-fei i, 0 'G, ll.
[1943] F3C'
[1944]
[1945] 2. n-BuLi, THF, -78 ‘C - ft, 2 h
[1946] To a solution of methyltriphenylphosphaniumbromide (1227.68 mg, 3.437 mmol, 1.3 equiv) in THF (5 mL) was added n-BuLi (220.15 mg, 3.437 mmol, 1.3 equiv) dropwise at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at 0 °C for 10 min under nitrogen atmosphere. To a stirred solution of aforementioned reaction system was added 1-(4-aminophenyl)-2,2,2-trifluoroethan-1-one (500 mg, 2.644 mmol, 1 equiv) in THF (5 mL) dropwise at -78 °C under nitrogen atmosphere. The resulting mixture was stirred at -78 °C for 30 mins under nitrogen atmosphere. Then, the reaction was allowed to proceed at room temperature. Desired product could be detected by GCMS. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 35 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (10 / 1) to afford 4-(3,3,3-trifluoroprop-1-en-2-yl)aniline as a yellow oil. GCMS (ESI) calcd. for CgHaFsN, 187.06 m / z, found 187.06[M]
[1947] Step 2. Synthesis of 4-(1,1,1-trifluoropropan-2-yl)aniline
[1948] Pd / C, MeOH FsC^^'i
[1949]
[1950] PATENT
[1951] ATTORNEY-DOCKET NO.: 51478-036WO4
[1952] To a solution of 4-(3,3,3-trifluoroprop-1-en-2-yl)aniline (100 mg, 0.534 mmol, 1 equiv) in MeOH (1 mL) was added Pd / C (56.86 mg, 0.534 mmol, 1 equiv) in portions at room temperature under hydrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h under hydrogen atmosphere. The reaction was monitored by GCMS. The resulting mixture was filtered, the filter cake was washed with MeOH (3x2 mL). The filtrate was concentrated under reduced pressure, to afford 4-(1,1,1-trifluoropropan-2-yl)aniline (95 mg, 93.99% yield, 90% purity) as a yellow oil. GCMS (ESI) calcd. for C9H10F3N, 189.08 m / z, found 189.08[M]
[1953] Step 3. Synthesis of 3-methyl-6-((R)-2-methylmorpholino)-8-((4-(1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1954] 6„ J
[1955] DIEA, DMSO
[1956] .11 xjL, N ji j <
[1957] Cf ' N Y 60 'C, 5 hr (■' " W' 'N' 'Vx
[1958] O o
[1959]
[1960] To a solution of 4-(1,1,1-trifluoropropan-2-yl)aniline (70 mg, 0.370 mmol, 1 equiv) and 6,8-dichloro-3-methylpyrido[3,2-d]pyrimidin-4(3H)-one (93.63 mg, 0.407 mmol, 1.1 equiv) in DMSO (1 mL) was added DIEA (143.47 mg, 1.110 mmol, 3 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 50 °C for 1 h under nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Add (2R)-2-methylmorpholine (41.17 mg, 0.407 mmol, 1.1 equiv) to above reaction system. The resulting mixture was stirred at 100 °C for 1 h under nitrogen atmosphere. The reaction was monitored by LCMS. Desired product could be detected by LCMS. The mixture was allowed to cool down to room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: (column, C18 silica gel; mobile phase, MeCN in water (0.1% TFA), 10% to 90% gradient in 20 min; detector, UV 254 nm) to afford 3-methyl-6-((R)-2-methylmorpholino)-8-((4-(1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (70 mg, 42.19% yield, 96% purity) as a white solid. MS (ESI) calcd. for C21H23F3N6O2, 448.18 m / z, found 449.25[M+H]+.
[1961] Step 4. Separation of 3-methyl-6-((R)-2-methylmorpholino)-8-((4-((R)-1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-methyl-6-((R)-2-methylmorpholino)-8-((4-((S)-1,1,1 -trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[1962]
[1963] The product (60 mg) was purified by Prep-HPLC with the following conditions (Column:
[1964] CHIRALPAK IK, 5 pm, 250 mm 20 mm; Mobile Phase A: Hex (with 0.5% NH3( 2 M in MeOH)), MobilePATENT
[1965] ATTORNEY-DOCKET NO.: 51478-036WO4
[1966] Phase B: (IPA: DCM=1: 1); Flow rate: 20 mL / min; Gradient (B%)30% B; Wave Length: 220 / 254 nm; Sample Solvent: ETOH; Injection Volume: 0.7 mL; Number Of Runs: 10; Single injection run time(min): 17.5) to afford
[1967] 3-methyl-6-((R)-2-methylmorpholino)-8-((4-((R)-1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (9.1 mg, 15.1% yield, 99.4% purity) as a white solid, stereochemistry arbitrarily assigned.
[1968] RT1(min): 14.08;
[1969] MS (ESI) calcd. for C21H23F3N6O2, 448.18 m / z, found 449.15[M+H]+
[1970] 1H-NMR (300 MHz, DMSO-d6) 6 (ppm): 9.64 (s, 1 H), 8.23 (s, 1 H), 7.86 - 7.99 (m, 2H), 7.32 - 7.45 (m, 2H), 4.36 - 4.59 (m, 2H), 3.85 - 3.98 (m, 1H), 3.67 - 3.84 (m, 1H), 3.43 - 3.58 (m, 5H), 2.91 - 3.08 (m, 1 H), 2.60 - 2.75 (m, 1 H), 1.39 - 1.52 (m, 3H), 1.10 - 1.20 (m, 3H).
[1971] And also to obtain 3-methyl-6-((R)-2-methylmorpholino)-8-((4-((S)-1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (9 mg, 15.0% yield, 99.5% purity) as a white solid, stereochemistry arbitrarily assigned.
[1972] RT2(min): 16.25;
[1973] MS (ESI) calcd. for C21H23F3N6O2, 448.18 m / z, found 449.10[M+H]+
[1974] 1H-NMR (300 MHz, DMSO-d6) 6 (ppm): 9.65 (s, 1 H), 8.23 (s, 1 H), 7.86 - 7.97 (m, 2H), 7.34 - 7.43 (m, 2H), 4.37 - 4.58 (m, 2H), 3.88 - 3.98 (m, 1H), 3.68 - 3.81 (m, 1H), 3.43 - 3.59 (m, 5H), 2.92 - 3.06 (m, 1 H), 2.59 - 2.75 (m, 1 H), 1.40 - 1.51 (m, 3H), 1.08 - 1.21 (m, 3H).
[1975] Example 97: (R)-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (Compound 100)
[1976] Step 1. Synthesis of 5-amino-2-chloro-6-methoxy-N-(tetrahydro-2H-pyran-4-yl) pyrimidine-4-carboxamide
[1977] - y
[1978] .: N. X 'V. MH2HATU. DSEA. DMF
[1979] yJL --A. OH
[1980] Of H' K ft, 1 hr
[1981]
[1982] 0
[1983] To a solution of 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid (4 g, 19.648 mmol, 1 equiv) and HATU (8965.16 mg, 23.578 mmol, 1.2 equiv) in DMF (40 mL) were added DIEA (7618.45 mg, 58.944 mmol, 3 equiv) and tetrahydro-2H-pyran-4-amine (2384.89 mg, 23.578 mmol, 1.2 equiv) at room temperature. The resulting mixture was stirred at room temperature for 1h. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 70% gradient in 20 min; detector, UV 254 nm. This resulted in 5-amino-2-chloro-6-methoxy-N-(tetrahydro-2H-pyran-4-yl) pyrimidine-4-carboxamide (4.2 g, 74.55% yield, 98% purity) as a yellow solid. MS: MS (ESI) calcd. for C11H15CIN4O3: 286.08, Found: 287.05 [M+H]+.PATENT
[1984] ATTORNEY-DOCKET NO.: 51478-036WO4
[1985] Step 2. Synthesis of 6-chloro-8-methoxy-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1986] "'Oc■ ‘ c
[1987] . NHaHOAc N- Ji A N A A, N. xx
[1988] Cl’ N y Y ■') 120’0,4 hrs N I T )
[1989]
[1990] 6 k.,,o 6 >, o
[1991] To a solution of 5-amino-2-chloro-6-methoxy-N-(tetrahydro-2H-pyran-4-yl) pyrimidine-4-carboxamide (1 g, 3.488 mmol, 1 equiv) in trimethoxymethane (10 mL) was added HOAc (1.05 g, 17.440 mmol, 5 equiv) dropwise at room temperature. The resulting mixture was stirred at 120°C for 4 h. The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in 6-chloro-8-methoxy-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (800 mg, 77.31% yield, 99% purity) as a white solid. MS: MS (ESI) calcd. for C12H13CIN4O3: 296.07, Found: 297.05 [M+H]+.
[1992] Step 3. Synthesis of (R)-8-methoxy-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[1993] 9 ‘" O
[1994] DJEA. OMSOuA..xN>..
[1995] - * j J
[1996] c N::: ■ 100’C, 1 hr r N N y y >
[1997] 0A. A 6.^ Y 6 Y,6
[1998]
[1999] To a solution of 6-chloro-8-methoxy-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (2700 mg, 9.100 mmol, 1 equiv) and (R)-2-methylmorpholine (1104.52 mg, 10.920 mmol, 1.2 equiv) in DMSO (30 mL) was added DIEA (2352.24 mg, 18.200 mmol, 2 equiv) dropwise at room temperature. The resulting mixture was stirred at 100°C for 1h. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 70% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-8-methoxy-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (2.5 g, 76.02% yield, 98% purity) as a white solid. MS: MS (ESI) calcd. for C17H23N5O4: 361.18, Found: 362.10 [M+H]+.
[2000] Step 4. Synthesis of (R)-8-hydroxy-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2001] O OH UCXTsaitDMF A
[2002] ..... X A. - *. 11 4N, f N N- Y Y - hr AAHA;
[2003] A J o k,6 6 J o k 0
[2004]
[2005] I I
[2006] To a solution of (R)-8-methoxy-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (2.5 g, 6.918 mmol, 1 equiv) and LiCI (1.46 g, 34.590 mmol, 5 equiv) in DMF (30PATENT
[2007] ATTORNEY-DOCKET NO.: 51478-036WO4
[2008] mL) was added TsOH (5.95 g, 34.590 mmol, 5 equiv) at room temperature. The resulting mixture was stirred at 120°C for 1h. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 1% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-8-hydroxy-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (2 g, 83.23% yield, 98% purity) as a white solid. MS: MS (ESI) calcd. for C16H21N5O4: 347.16, Found: 348.10 [M+H]+.
[2009] Step 5. Synthesis of (R)-8-chloro-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2010] OH a
[2011] NAAPC°-> NAA
[2012] f / A 7 «A IfA T - } 8O’C< 1 hr * f Jl Hl y N V
[2013] 0^ 0 k,a o. J a I.6
[2014]
[2015] I - I
[2016] A solution of (R)-8-hydroxy-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (2.0 g, 5.757 mmol, 1 equiv) in POCl3 (20 mL) was stirred at 80°C for 1h. The resulting mixture was concentrated under vacuum. The reaction was quenched with ice water at 0°C. The mixture was neutralized to pH 8 with saturated NaHCCh (aq.). The resulting mixture was extracted with EtOAc (3 x 50 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in (R)-8-chloro-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (1.8 g, crude) as a yellow solid. MS: MS (ESI) calcd. for C16H20CIN5O3: 365.13, Found: 366.10 [M+H]+.
[2017] Step 6. Synthesis of (R)-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2018] N Y % DSEA. DMSO
[2019] .-SI A N.
[2020] M |f -r; go's, 1 hr
[2021] o. J o L 0
[2022]
[2023] I
[2024] To a solution of (R)-8-chloro-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (300 mg, 0.820 mmol, 1 equiv) and 4-isopropylaniline (133.06 mg, 0.984 mmol, 1.2 equiv) in DMSO (5 mL) was added DIEA (211.98 mg, 1.640 mmol, 2 equiv) dropwise at room temperature. The resulting mixture was stirred at 50°C for 1h. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 90% gradient in 30 min; detector, UV 254 nm. This resulted in (R)-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (316.2 mg, 83.00% yield, 99.7% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C25H32N6O3: 464.25. Found: 465.20 [M+H]+.1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 9.49 (s, 1H), 8.25 (s, 1H), 7.77 - 7.79 (m, 2H), 7.22 - 7.24 (m, 2H), 4.80 - 4.86 (m, 1H), 4.41 - 4.52 (m, 2H), 3.99 - 4.03 (m, 2H), 3.89 - 3.93 (m, 1H), 3.51 - 3.52 (m, 2H), 3.47 - 3.49 (m, 2H), 2.96 - 3.02 (m, 1H), 2.85 - 2.89 (m, 1H), 2.63 - 2.68 (m, 1H), 2.07 - 2.16 (m, 2H), 1.76 - 1.79 (m, 2H), 1.15 - 1.21 (m, 9H).PATENT
[2025] ATTORNEY-DOCKET NO.: 51478-036WO4
[2026] Using the approach to synthesize Example 97, the compounds of Table 7 were synthesized from the corresponding amine.
[2027] Table 7: Experimental Data for Compounds
[2028] Ex Cmpd
[2029] Name Structure MS 1H NMR
[2030] # #
[2031] 98 101 8-((4-isopropylphenyl)- 451.15 1H NMR (300 MHz,
[2032] amino)-6-((R)-2- [M+H]+DMSO-d6) 6 (ppm): 8.11 - methylmorpholino)-3- 8.12 (m, 1H), 7.73 - 7.76 ((R)-tetrahydrofuran-3- (m, 2H), 7.25 - 7.28 (m, yl)pyrimido[5,4- 2H), 5.30 - 5.31 (m, 1H), d]pyrimidin-4(3H)-one 4.38 - 4.50 (m, 2H), 4.00 - 4.11 (m, 2H), 3.81 - 3.91 O. J O (m, 3H), 3.46 - 3.66 (m, I
[2033] 2H), 2.91 - 3.04 (m, 1H), 2.75 - 2.86 (m, 1H), 2.53 - 2.67 (m, 1H), 2.46 - 2.48 (m, 1H), 2.14 - 2.18 (m, 1H), 1.15 - 1.22 (m, 9H). 99 102 8-((4-isopropylphenyl)- 451.15 1H NMR (300 MHz,
[2034] amino)-6-((R)-2- [M+H]+DMSO-d6) 6: 8.12 (s, 1H), methylmorpholino)-3- 7.77 - 7.79 (m, 2H), 7.27 - ((S)-tetrahydrofuran-3- 7.28 (m, 2H), 5.31 (s, 1H), yl)pyrimido[5,4- 4.39 - 4.50 (m, 2H), 4.01 - d]pyrimidin-4(3H)-one 4.14 (m, 2H), 3.78 - 3.93 AA
[2035] (m, 3H), 3.47 - 3.54 (m, i 2H), 2.91 - 3.10 (m, 1H),
[2036] 2.76 - 2.89 (m, 1H), 2.73 - 2.76 (m, 1H), 2.53 - 2.55 (m, 1H), 2.16 - 2.34 (m, 1H), 1.19 - 1.28 (m, 9H). 100 103 (R)-3-(2,2-difluoroethyl)- 445.25 1H NMR (300 MHz,
[2037] 8-((4-isopropylphenyl)- [M+H]+MeOD) 6 8.15 (s, 1H), amino)-6-(2-methyl- 7.66 - 7.69 (m, 2H), 7.25 - morpholino)pyrimido[5,4- 7.28 (m, 2H), 6.08 - 6.27 d]pyrimidin-4(3H)-one (m, 1H), 4.44 - 4.64 (m, N A
[2038] 4H), 3.94 - 3.99 (m, 1H), f N A 'hr -A. yN >
[2039] ex J 6 A,,, 3.57 - 3.66 (m, 2H), 3.05 - rF F
[2040] 3.28 (m, 1H), 2.83 - 3.00 (m, 1H), 2.70 - 2.83 (m, 1H), 1.24 - 1.33 (m, 9H).
[2041]
[2042] PATENT
[2043] ATTORNEY-DOCKET NO.: 51478-036WO4
[2044] 101 104 8-((4-isopropylphenyl)- 464.25 1 H NMR (500 MHz,
[2045] amino)-6-((R)-2- [M+H]+MeOD) 6 8.21 (s, 1 H), methylmorpholino)-3- 7.68 - 7.69 (m, 2H), 7.30 - ((R)-1 -methylpyrrolidin- 7.32 (m, 2H), 5.23 (s, 1 H), 3-yl)pyrimido[5,4- 4.51 - 4.58 (m, 2H), 4.09 - d]pyrimidin-4(3H)-one 4.11 (m, 2H), 4.00 - 4.03
[2046] (m, 1 H), 3.63 - 3.67 (m, 3H), 3.20 - 3.23 (m, 1 H), I 3.10 - 3.15 (m, 1 H), 3.07
[2047] (s, 3H), 2.96 - 2.98 (m, 1 H), 2.84 - 2.89 (m, 1 H), 2.76 - 2.80 (m, 1 H), 2.56 - 2.60 (m, 1 H), 1.29 - 1.32 (m, 6H), 1.26 - 1.28 (m, 3H).
[2048] 102 105 8-((4-isopropylphenyl)- 464.35 1 H NMR (500 MHz,
[2049] amino)-6-((R)-2- [M+H]+MeOD) 6 8.09 (s, 1 H), methylmorpholino)-3- 7.70 - 7.72 (m, 2H), 7.27 - ((S)-1-methylpyrrolidin-3- 7.29 (m, 2H), 5.16 - 5.18 yl)pyrimido[5,4- (m, 1 H), 4.70 - 4.87 (m, d]pyrimidin-4(3H)-one 2H), 4.01 - 4.13 (m, 2H),
[2050] 3.98 - 4.00 (m, 1 H), 3.60 - 3.66 (m, 3H), 3.33 - 3.36 (m, 1 H), 3.28 - 3.32 (m, 2H), 3.06 - 3.10 (m, 3H), 2.92 - 2.96 (m, 1 H), 2.75 - 2.79 (m, 1 H), 2.70 - 2.73 (m, 1 H), 1.24 - 1.29 (m, 9H).
[2051] 103 106 6-(diethylamino)-3- 463.15 1 H NMR (400 MHz,
[2052] (tetrahydro-2H-pyran-4- [M+H]+DMSO-d6) 6 (ppm): 9.91 yl)-8-((4- (s, 1 H), 8.20 - 8.25 (m, (trifluoromethyl)phenyl)a 3H), 7.70 - 7.73 (m, 2H), mino)pyrimido[5,4- 4.80 - 4.86 (m, 1 H), 4.00 - d]pyrimidin-4(3H)-one 4.04 (m, 2H), 3.47 - 3.52
[2053] (m, 4H), 3.34 - 3.39 (m, 2H), 2.32 - 2.34 (m, 2H), 1.77 - 1.80 (m, 2H), 1.17 - 1.19 (m, 6H).
[2054]
[2055] PATENT
[2056] ATTORNEY-DOCKET NO.: 51478-036WO4
[2057] 104 107 6-(dimethylamino)-3- 435.10 1H NMR (400 MHz,
[2058] (tetrahydro-2H-pyran-4- [M+H]+DMSO-d6) 6 (ppm): 9.91 yl)-8-((4- (s, 1H), 8.20 - 8.25 (m, (trifluoromethyl)phenyl)a 3H), 7.71 - 7.73 (m, 2H), mino)pyrimido[5,4- 4.82 - 4.87 (m, 1H), 4.00 - d]pyrimidin-4(3H)-one 4.04 (m, 2H), 3.47 - 3.53
[2059] (m, 2H), 3.11 - 3.19 (m, 6H), 2.18 - 2.21 (m, 2H), 1.78 - 1.81 (m, 2H).
[2060] 105 108 (R)-6-(2- 491.20 1H NMR (400 MHz,
[2061] methylmorpholino)-3- [M+H]+DMSO-d6) 6 (ppm): 10.02 (tetrahydro-2H-pyran-4- (s, 1H), 8.28 (s, 1H), 8.13 yl)-8-((4- - 8.15 (m, 2H), 7.72 - 7.74 (trifluoromethyl)phenyl)a (m, 2H), 4.81 - 4.87 (m, mino)pyrimido[5,4- k ii 1H), 4.47 - 4.51 (m, 2H),,1 N.
[2062] d]pyrimidin-4(3H)-one I 'TV'1. 4.00 - 4.03 (m, 2H), 3.91 - ’’'A'"'';
[2063] o. > o <,6 3.94 (m, 1H), 3.47 - 3.55 I
[2064] (m, 4H), 2.99 - 3.04 (m, 1H), 2.66 - 2.72 (m, 1H), 2.12 - 2.18 (m, 2H), 1.77 - 1.81 (m, 2H), 1.17 - 1.19 (m, 3H).
[2065]
[2066] Example 106: 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-((S)-1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-((R)-1,1,1 -trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 109 and 110)
[2067] Step 1. Synthesis of 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-(1,1,1-trifluoropropan-2-yl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2068] C!
[2069] EHEA. OMSO
[2070] JI N *"..
[2071] f N N y r 1 we, a nr r
[2072] 6. J d k.o □ J
[2073]
[2074] i ' Y
[2075] To a solution of (R)-8-chloro-6-(2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl) pyrimido[5,4-d] pyrimidin-4(3H)-one (200 mg, 0.547 mmol, 1 equiv) and para-isopropylaniline (133.0 mg, 0.984 mmol, 1.2 equiv) in DMSO (5 mL) was added DIEA (212.0 mg, 1.641 mmol, 3 equiv) dropwise at room temperature. The resulting mixture was stirred at 100°C for 3h. LCMS showed the reaction was completed, the residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 90% gradient in 30 min; detector, UV 254 nm. ThisPATENT
[2076] ATTORNEY-DOCKET NO.: 51478-036WO4
[2077] resulted in 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-(1,1,1-trifluoropropan-2-yl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (316.2 mg, 83.00% yield, 99.7% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C25H29F3N6O3: 518.23, Found: 519.05[M+H]+.
[2078] Step 2. Separation of 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-((S)-1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-((R)-1,1,1 -trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[2079]
[2080] The product (270 mg) was separated by chiral HPLC with the following conditions: UniChiral OD-5H 2*25 cm, 5 pm; Mobile Phase A: Hex (with 0.1% DEA), Mobile Phase B: ETOH; Flow rate: 20 mL / min; Gradient (B%)30% B; Wave Length: 220 / 254 nm; Sample Solvent: EtOH--HPLC; Injection Volume: 0.5 mL; Number Of Runs: 24; Single injection run time(min): 25 to afford 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-((S)-1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (51.0 mg, 15.09% yield, 99.1% purity) as a yellow solid,
[2081] RT1(min): 16.34
[2082] LC / MS: MS (ESI) calcd. for C25H29F3N6O3: 518.54. Found: 519.10[M+H]+.
[2083] 1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 9.42 (s, 1 H), 8.28 (s, 1 H), 7.88 - 7.91 (m, 2H), 7.38 - 7.40 (m, 2H), 4.80 - 4.86 (m, 1H), 4.40 - 4.51 (m, 2H), 4.00 - 4.03 (m, 2H), 3.90 - 3.94 (m, 1H), 3.74 - 3.78 (m, 1H), 3.47 - 3.54 (m, 4H), 2.98 - 3.03 (m, 1H), 2.65 - 2.70 (m, 1H), 2.13 - 2.17 (m, 2H), 1.77 - 1.80 (m, 2H), 1.44 - 1.45 (m, 3H), 1.15 - 1.17 (m, 3H).
[2084] And also to afford 6-((R)-2-methylmorpholino)-3-(tetrahydro-2H-pyran-4-yl)-8-((4-((R)-1,1,1-trifluoropropan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (38.9 mg, 11.51% yield, 98.9% purity) as a yellow solid,
[2085] RT2(min): 20.78.
[2086] LC / MS: MS (ESI) calcd. for C25H29F3N6O3: 518.54. Found: 519.15[M+H]+.
[2087] 1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 9.75 (s, 1 H), 8.28 (s, 1 H), 7.88 - 7.90 (m, 2H), 7.38 - 7.40 (m, 2H), 4.80 - 4.86 (m, 1H), 4.41 - 4.51 (m, 2H), 4.00 - 4.03 (m, 2H), 3.90 - 3.94 (m, 1H), 3.74 - 3.78 (m, 1H), 3.47 - 3.54 (m, 4H), 2.98 - 3.04 (m, 1H), 2.65 - 2.71 (m, 1H), 2.13 - 2.17 (m, 2H), 1.77 - 1.80 (m, 2H), 1.44 - 1.45 (m, 3H), 1.15 - 1.17 (m, 3H).PATENT
[2088] ATTORNEY-DOCKET NO.: 51478-036WO4
[2089] Example 107: (R)-2-(difluoromethyl)-3-ethyl-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (Compound 111)
[2090] Step 1. Synthesis of (R)-5-Amino-N-ethyl-6-methoxy-2-(2-methylmorpholino) pyrimidine-4-carboxamide
[2091] NM Q V I O
[2092] . A. HH-s 3 eMoropiytidlne, CssCO3. <£oxac«., A- / NHJ
[2093] N "f: uNT u ^4 A. tsj..... H cr s' y wc,zhra f 'H' N' Y d i o, o!
[2094]
[2095] To a solution of 5-amino-2-chloro-N-ethyl-6-methoxypyrimidine-4-carboxamide (1.5 g, 6.080 mmol, 1 equiv) and (R)-2-methylmorpholine (0.74 g, 7.296 mmol, 1.2 equiv) in Dioxane (10 mL) were added Pd-PEPPSI-IHeptCI 3-chloropyridine (888.10 mg, 0.912 mmol, 0.15 equiv) and Cs₂CO₃ (5.942 g, 18.240 mmol, 3 equiv) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100°C for 2 h under nitrogen atmosphere. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-5-Amino-N-ethyl-6-methoxy-2-(2-methylmorpholino) pyrimidine-4-carboxamide (1.2 g, 63.38% yield, 98% purity) as a white solid. MS: MS (ESI) calcd. for C13H21N5O3: 295.16, Found: 296.10 [M+H]+.
[2096] Step 2. Synthesis of (R)-5-(2,2-difluoroacetamido)-N-ethyl-6-methoxy-2-(2-methylmorpholino) pyrimidine-4-carboxamide
[2097] o o ■
[2098] N n
[2099]
[2100] To a solution of (R)-5-Amino-N-ethyl-6-methoxy-2-(2-methylmorpholino) pyrimidine-4-carboxamide (700 mg, 2.370 mmol, 1 equiv) and 2,2-difluoroacetic anhydride (495.03 mg, 2.844 mmol, 1.2 equiv) in DCM (10 mL) was added TEA (479.68 mg, 4.740 mmol, 2 equiv) dropwise at 0°C. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 70% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-5-(2,2-difluoroacetamido)-N-ethyl-6-methoxy-2-(2-methylmorpholino) pyrimidine-4-carboxamide (400 mg, 45.20% yield, 97% purity) as a brown oil. MS: MS (ESI) calcd. for C15H21F2N5O4: 373.16, Found: 374.10 [M+H]+.PATENT
[2101] ATTORNEY-DOCKET NO.: 51478-036WO4
[2102] Step 3. Synthesis of (R)-2-(difluoromethyl)-3-ethyl-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2103]
[2104] A solution of (R)-5-(2,2-difluoroacetamido)-N-ethyl-6-methoxy-2-(2-methylmorpholino) pyrimidine-4-carboxamide (500 mg, 1.339 mmol, 1 equiv) in HOAc (10 mL) was stirred at 130°C for 4 h. The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 70% gradient in 30 min; detector, UV 254 nm. This resulted in (R)-2-(difluoromethyl)-3-ethyl-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (350 mg, 73.55% yield, 96% purity) as a white solid. MS: MS (ESI) calcd. for C15H19F2N5O3: 355.15, Found: 356.10 [M+H]+.
[2105] Step 4. Synthesis of (R)-2-(difluoromethyl)-3-ethyl-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2106] OH F
[2107] Av A,
[2108] LiCl. TsOH. DWSO N 'y ' YF....................................... fc..... Af, J <-A 'y N >
[2109] 1:20'C, 1 Sir 6. 6!
[2110]
[2111] I
[2112] To a solution of (R)-2-(difluoromethyl)-3-ethyl-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (700 mg, 1.970 mmol, 1 equiv) and TsOH (1696.09 mg, 9.850 mmol, 5 equiv) in DMF (10 mL) was added LiCI (417.52 mg, 9.850 mmol, 5 equiv) at room temperature. The resulting mixture was stirred at 120°C for 1h. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 50% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-2-(difluoromethyl)-3-ethyl-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (600 mg, 89.24% yield, 97% purity) as a white solid. MS: MS (ESI) calcd. for C14H17F2N5O3: 341.13, Found: 342.10 [M+H]+.
[2113] Step 5. Synthesis of (R)-8-chloro-2-(difluoromethyl)-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2114] 80'0. 1 hr
[2115]
[2116] A solution of (R)-2-(difluoromethyl)-3-ethyl-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (600 mg, 1.758 mmol, 1 equiv) in POCl3 (10 mL) was stirred at 80°C for 1h. The resulting mixture was concentrated under vacuum. The reaction was quenched with ice water at 0°C. The mixture was neutralized to pH 7 with saturated NaHCCh (aq.). The resulting mixture was extracted with EtOAc (3 x50 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated underPATENT
[2117] ATTORNEY-DOCKET NO.: 51478-036WO4
[2118] reduced pressure. This resulted in (R)-8-chloro-2-(difluoromethyl)-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (450 mg, crude) as a yellow solid. MS: MS (ESI) calcd. for C14H16CIF2N5O2: 359.10, Found: 360.10 [M+H]+.
[2119] Step 6. Synthesis of (R)-2-(difluoromethyl)-3-ethyl-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2120] S F L X..1.
[2121] I N X T
[2122] N— T A A A
[2123] I J A OffiA, DMSO Ijl 'Y V **
[2124] A A. N.
[2125] 6V) 6 * wc„ 1 hr fNf 1 | o.. 0!
[2126]
[2127] To a solution of (R)-8-chloro-2-(difluoromethyl)-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (150 mg, 0.417 mmol, 1 equiv) and 4-isopropylaniline (67.65 mg, 0.500 mmol, 1.20 equiv) in DMSO (5 mL) was added DIEA (107.78 mg, 0.834 mmol, 2 equiv) at room temperature. The resulting mixture was stirred at 100°C for 1h. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 2% to 80% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-2-(difluoromethyl)-3-ethyl-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (134.7 mg, 56.43% yield, 99.4% purity) as a trifluoroacetate salt and as a light yellow solid. LC / MS: MS (ESI) calcd. for C23H28F2N6O2: 458.24. Found: 459.15 [M+H]+.1H NMR (300 MHz, DMSO-d₆) 6 (ppm): 7.76 - 7.77 (m, 2H), 7.37 - 7.40 (m, 2H), 6.84 -7.28 (m, 1H), 4.43 - 4.56 (m, 2H), 4.09 - 4.16 (m, 2H), 3.90 - 3.94 (m, 1H), 3.48 - 3.52 (m, 2H), 3.03 - 3.07 (m, 1H), 2.84 - 2.99 (m, 1H), 2.65 - 2.73 (m, 1H), 1.27 - 1.30 (m, 3H), 1.19 - 1.26 (m, 9H).
[2128] Using the approach in Example 107, the compounds of Table 8 were synthesized from the corresponding amine and the corresponding aniline.PATENT
[2129] ATTORNEY-DOCKET NO.: 51478-036WO4
[2130] Table 8: Experimental Data for Compounds
[2131] Ex Cmpd
[2132] Name Structure MS 1H NMR
[2133] # #
[2134] 108 112 8-((4-(3,3-difluorocyclo- 549.15 1 H NMR (300 MHz, DMSO- butyl)phenyl)amino)-2- [M+H]+d6) 6: 9.38 (s, 1 H), 7.80 - (difluoromethyl)-6-((R)- 7.83 (m, 2H), 7.30 - 7.42 2-methylmorpholino)-3- (m, 2H), 6.89 - 7.08 (m, 1 H), ((R)-tetrahydrofuran-3- 4.90 - 5.08 (m, 1 H), 4.32 - NH F
[2135] yl)pyrimido[5,4- 4.61 (m, 2H), 4.10 - 4.23. J! J. N
[2136] d]pyrimidin-4(3H)-one 6 J d I.? (m, 1 H), 3.85 - 4.02 (m, 5H),
[2137] J
[2138] 3.30 - 3.42 (m, 2H), 2.88 - 3.15 (m, 3H), 2.62 - 2.81 (m, 3H), 2.32 - 2.45 (m, 1 H), 2.10 - 2.28 (m, 1 H), 1.08 - 1.16 (m, 3H).
[2139] 109 113 8-((4-(3,3- 549.15 1 H NMR (300 MHz, DMSO- difluorocyclobutyl)phen [M+H]+d6) 6 (ppm): 7.80 - 7.82 (m, yl)amino)-2- 2H), 7.33 - 7.35 (m, 2H), (difluoromethyl)-6-((R)- 6.89 - 7.25 (m, 1 H), 4.82 - 2-methylmorpholino)-3- 5.11 (m, 1 H), 4.32 - 4.75 ((S)-tetrahydrofuran-3- (m, 2H), 4.08 - 4.26 (m, 1 H),
[2140] . A A A x
[2141] yl)pyrimido[5,4- O r >? « x T o 3.89 - 4.02 (m, 4H), 3.61 - d]pyrimidin-4(3H)-one 3.62 (m, 1 H), 3.35 - 3.43 (m, 2H), 2.91 - 3.11 (m, 3H), 2.63 - 2.78 (m, 3H), 2.35 - 2.43 (m, 1 H), 2.11 - 2.27 (m, 1 H), 1.10 - 1.21 (m, 3H).
[2142] 110 114 (R)-2-(difluoromethyl)- 447.25 1 H NMR (500 MHz, MeOD):
[2143] 3-ethyl-8-((4-methoxy- [M+H]+6 7.65 - 7.66 (m, 2H), 6.87 - phenyl)amino)-6-(2- 7.01 (m, 3H), 4.53 - 4.64 methylmorpholino)pyri F (m, 2H), 4.32 - 4.36 (m, 2H), mido[5,4-d]pyrimidin- 3.95 - 4.02 (m, 1 H), 3.84 (s,
[2144] JI J «
[2145] 4(3H)-one f Y N Y J
[2146] J o!3H), 3.57 - 3.68 (m, 2H), 1 3.16 - 3.19 (m, 1 H), 2.80 - 2.84 (m, 1 H), 1.43 - 1.44 (m, 3H), 1.24 - 1.25 (m, 3H).
[2147]
[2148] PATENT
[2149] ATTORNEY-DOCKET NO.: 51478-036WO4
[2150] 111 115 8-((3-chlorophenyl)- 437.15 1 H NMR (500 MHz, MeOD) amino)-2-(difluoro- [M+H]+6 8.01 - 8.02 (m, 1 H), 7.68 - methyl)-3-ethyl-6- 7.70 (m, 1 H), 7.37 - 7.40 morpholinopyrimido[5,4 (m, 1 H), 7.16 - 7.18 (m, 1 H),
[2151] H X. Y P-L y, L r
[2152] -d]pyrimidin-4(3H)-one A -A.. N 6.78 - 6.99 (m, 1 H), 4.30 - p" W N 'jf -[
[2153] O 4.35 (m, 2H), 3.92 - 3.94 (m, 4H), 3.77 - 3.79 (m, 4H), 1.42 - 1.44 (m, 3H).
[2154] 112 116 (R)-2-(difluoromethyl)- 447.25 1 H NMR (400 MHz, MeOD)
[2155] 8-((4-isopropylphenyl)- [M+H]+6 7.65 - 7.68 (m, 2H), 7.27 - amino)-3-methyl-6-(2- 7.29 (m, 2H), 6.73 - 6.99 methylmorpholino)pyri (m, 1 H), 4.62 - 4.65 (m, 2H), mido[5,4-d]pyrimidin- F 3.95 - 4.03 (m, 1 H), 3.78 - N’ A xy;-i yA. -f
[2156] 4(3H)-one..... Al. J...
[2157] < N' y -• 3.80 (m, 3H), 3.58 - 3.63 (m, 2H), 3.16 - 3.18 (m, 1 H), 2.88 - 3.01 (m, 1 H), 2.81 - 2.83 (m, 1 H), 1.27 - 1.30 (m, 9H).
[2158] 113 117 (R)-2-(difluoromethyl)- 453.10 1 H NMR (400 MHz, DMSO- 8-((4-(difluoromethyl)- [M+H]+d6) 6 (ppm): 7.92 - 8.06 (m, phenyl)amino)-3- F 2H), 7.56 - 7.82 (m, 2H), methyl-6-(2-methyl- 6.81 - 7.18 (m, 2H), 4.27 - F
[2159] morpholino)pyrimido[5, N A F 4.68 (m, 2H), 3.89 - 4.00 AL
[2160] 4-d p r m d f N M Y
[2161] ] y i i in-4(3H)- ' if
[2162] D J 6 (m, 1 H), 3.65 - 3.67 (m, 4H), one I 3.38 - 3.40 (m, 1 H), 2.92 - 3.12 (m, 1 H), 1.11 - 1.28 (m, 1 H), 1.21 - 1.22 (m, 3H).
[2163] 114 118 2-(difluoromethyl)-8- 517.15 1 H NMR (300 MHz, DMSO- ((4-(1-fluorocyclo- [M+H]+d6) 6: 7.85 - 7.87 (m, 2H), propyl)phenyl)amino)- 7.32 - 7.35 (m, 2H), 6.90 - 6-((R)-2-methyl- 7.26 (m, 1 H),4.90 - 5.10 (m, morpholino)-3-((R)- Vi 1 H), 4.30 - 4.62 (m, 2H), MB F
[2164] tetrahydrofuran-3-. N,,-L,y. N -.. J,-A. F 4.10 - 4.18 (m, 1 H), 3.85 - yl)pyrimido[5,4- I T * T >■ C
[2165] o., / 0 L z 4.02 (m, 4H), 3.40 - 3.58 T
[2166] d]pyrimidin-4(3H)-one (m, 2H), 2.95 - 3.10 (m, 1 H),
[2167] 2.64 - 2.82 (m, 1 H), 2.32 - 2.48 (m, 1 H), 2.03 - 2.28 (m, 1 H), 1.40 - 1.51 (m, 2H), 1.10 - 1.18 (m, 5H).
[2168]
[2169] PATENT
[2170] ATTORNEY-DOCKET NO.: 51478-036WO4
[2171] 115 119 2-(difluoromethyl)-8- 517.15 1 H NMR (400 MHz, DMSO- ((4-(1-fluorocyclo- [M+H]+d6) 6 (ppm): 7.85 - 7.87 (m, propyl)phenyl)amino)- 2H), 7.32 - 7.34 (m, 2H), 6-((R)-2-methyl- 6.90 - 7.25 (m, 1 H), 4.95 - morpholino)-3-((S)- 5.07 (m, 1 H), 4.38 - 4.55 tetrahydrofuran-3- (m, 2H), 4.07 - 4.21 (m, 1 H), yl)pyrimido[5,4- 3.82 - 4.02 (m, 4H), 3.45 - M
[2172] d]pyrimidin-4(3H)-one 3.55 (m, 2H), 2.95 - 3.1 1 (m, 1 H), 2.65 - 2.75 (m, 1 H), 2.32 - 2.42 (m, 1 H), 2.11 - 2.21 (m, 1 H), 1.40 - 1.49 (m, 2H), 1.10 - 1.25 (m, 5H).
[2173] 116 120 2-(difluoromethyl)-8- 514.20 1 H NMR (400 MHz, DMSO- ((4-isopropylphenyl)- [M+H]+d6) 6 (ppm): 7.81 - 7.89 (m, amino)-6-((R)-2- 2H), 7.23 - 7.27 (m, 2H), methylmorpholino)-3- 6.92 - 7.18 (m, 1 H), 5.32 - ((R)-1 -methylpyrrolidin- 5.36 (m, 1 H), 4.46 - 4.59 3-yl)pyrimido[5,4- (m, 2H), 4.04 - 4.07 (m, 1 H), d]pyrimidin-4(3H)-one 3.91 - 3.99 (m, 2H), 3.55 - 3.61 (m, 2H), 3.30 - 3.32 (m, 1 H), 3.21 - 3.24 (m, 1 H), 3.01 - 3.08 (m, 2H), 2.87 - 3.08 (m, 4H), 2.68 - 2.74 (m, 1 H), 2.34 - 2.44 (m, 1 H), 1.20 - 1.22 (m, 6H), 1.14 - 1.16 (m, 3H).
[2174] 117 121 (R)-2-(difluoromethyl)- 471.05 1 H NMR (300 MHz, DMSO)
[2175] 3-methyl-6-(2- [M+H]+6: 9.71 (s, 1 H), 8.08 - 8.11 methylmorpholino)-8- (m, 2H), 7.74 - 7.77 (m, 2H), ((4-(trifluoromethyl)- 6.98 - 7.11 (m, 1 H), 4.46 - phenyl)amino)pyrimido[ 4.50 (m, 2H), 3.94 - 3.95 5,4-d]pyrimidin-4(3H)- (m, 1 H), 3.56 - 3.59 (m, 3H),
[2176] T
[2177] one 3.48 - 3.56 (m, 2H), 2.94 - 3.05 (m, 1 H), 2.71 - 2.73 (m, 1 H), 1.14 - 1.18 (m, 3H).
[2178]
[2179] PATENT
[2180] ATTORNEY-DOCKET NO.: 51478-036WO4
[2181] Example 118: (R)-8-((4-(3,3-difluorocyclobutyl)-3-fluorophenyl)amino)-3-methyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 122)
[2182] Step 1. Synthesis of 5-amino-2-chloro-N-methyl-6-(methylthio) pyrimidine-4-carboxamide
[2183] H2
[2184] "'S ' '"s
[2185] X. NHs MATU DlEAhDMFkX
[2186] X, < XNCL
[2187]
[2188] N rt, I hr Cl' N
[2189] 0 0
[2190] To a stirred solution of 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylic acid (2 g, 9.106 mmol, 1 equiv) and methanamine, hydrochloride (737.79 mg, 10.927 mmol, 1.2 equiv) in DMF (50 mL) were added HATU (6924.69 mg, 18.212 mmol, 2 equiv) and DIEA (3530.70 mg, 27.318 mmol, 3 equiv) at 0°C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1h under nitrogen atmosphere. LCMS showed that the desired product was formed. The reaction was quenched with saturated NH4CI (15 mL). The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in 5-amino-2-chloro-N-methyl-6-(methylthio) pyrimidine-4-carboxamide (2.1 g, 99.11% yield, 95% purity) as a yellow solid. LC / MS: MS (ESI) calcd for C7H9CIN4OS: 232.02. Found: 233.10[M+H]+
[2191] Step 2. Synthesis of 6-chloro-3-methyl-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2192] I
[2193] „X NHj HO*C
[2194] T H >
[2195] X, X N, X. kk N QF PF y 120X 6 hrs CT ’N" y
[2196]
[2197] O O
[2198] To a stirred solution of 5-amino-2-chloro-N-methyl-6-(methylthio) pyrimidine-4-carboxamide (1 g, 4.298 mmol, 1 equiv) in trimethoxymethane (6 mL) was added HOAc (774.23 mg, 12.894 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 120°C for 6h under nitrogen atmosphere. LCMS showed that the desired product was formed. The reaction mixture was quenched by addition of saturated NH4CI (20 mL). The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in 6-chloro-3-methyl-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (330 mg, 31.64% yield, 96% purity) as a white solid. LC / MS: MS (ESI) calcd for C8H7CIN4OS: 242.68. Found: 243.10[M+H]+.PATENT
[2199] ATTORNEY-DOCKET NO.: 51478-036WO4
[2200] Step 3. Synthesis of (R)-3-methyl-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2201] o.,>
[2202] ■S Y
[2203] OIEA. DMSQ
[2204] « I i N V
[2205] ..... J.( J. K
[2206] u” 8 WC, 1: ftr f N’ N' Y
[2207]
[2208] To a stirred solution of 6-chloro-3-methyl-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.412 mmol, 1 equiv) and (2R)-2-methylmorpholine (50.02 mg, 0.494 mmol, 1.2 equiv) in DMSO (4 mL) was added DIEA (106.52 mg, 0.824 mmol, 2 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100°C for 1 h under nitrogen atmosphere. LCMS showed desired product was formed. The reaction mixture was quenched by addition of saturated NH4CI (20 mL). The aqueous layer was extracted with EtOAc (3 x 100 mL). The combined organic phase was washed with brine (60 mL), dried over anhydrous Na2SO4. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in (R)-3-methyl-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (80 mg, 63.16% yield, 95% purity) as a yellow solid. LC / MS: MS (ESI) calcd for C13H17N5O2S: 307.11. Found: 308.10[M+H]+.
[2209] Step 4. Synthesis of 3-methyl-6-((R)-2-methylmorpholino)-8-(methylsulfinyl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2210]
[2211] To a stirred solution of (R)-3-methyl-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 0.195 mmol, 1 equiv) in DCM (3 mL) was added m-CPBA (67.37 mg, 0.390 mmol, 2 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred at 0°C for 1h under nitrogen atmosphere. After completion of the reaction. The resulting mixture was diluted with DCM (10 mL) and quenched with saturated NaHCOs (aq.) at 0°C. The aqueous layer was extracted with DCM (30 mL x 3). The combined organic phase was washed with sodium bicarbonate aqueous solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. This resulted in 3-methyl-6-((R)-2-methylmorpholino)-8-(methylsulfinyl) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, crude product) as a yellow solid. LC / MS: MS (ESI) calcd for C13H17N5O3S: 323.11. Found: 324.10[M+H]+.PATENT
[2212] ATTORNEY-DOCKET NO.: 51478-036WO4
[2213] Step 5. Synthesis of tert-butyl (4-(3,3-difluorocyclobutyl)-3-fluorophenyl) carbamate
[2214] “ ‘ OH j.
[2215] F F-VTh
[2216] Brj 1. NRC, pyritta. tSuOMe, rt, 15mm.- VQ
[2217] ' < O > - | J II p '‘“'-n A 2, K®r2. dtbbfjy. ''N " '" Q '' pwqymiscficSiEie, ph&ssfimide, JBisOMe / H
[2218]
[2219] DMA.. bk® LEDs, rt, 1Shra
[2220] To a stirred solution of 3,3-difluorocyclobutan-1-ol (285.00 mg, 2.637 mmol, 2.55 equiv) in MTBE (32.1 mL) was added 5,7-di-tert-butyl-3-phenylbenzo[d]oxazol-3-ium tetra fluoroborate (980.86 mg, 2.482 mmol, 2.4 equiv) at 0°C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 10min under nitrogen atmosphere. To this suspension, a solution of pyridine (196.30 mg, 2.482 mmol, 2.4 equiv) in MTBE (2.1 mL) was added dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 15min under nitrogen atmosphere. Another dram vial was charged with tert-butyl (4-bromo-3-fluorophenyl) carbamate (300 mg, 1.034 mmol, 1 equiv), NiBr2(dtbbpy) (25.17 mg, 0.052 mmol, 0.05 equiv), [lr(dtbbpy)(ppy)2]PFe (14.18 mg, 0.016 mmol, 0.015 equiv), Quinuclidine (201.20 mg, 1.810 mmol, 1.75 equiv) and Phthalimide (33.47 mg, 0.227 mmol, 0.22 equiv). DMAc (14.2 mL) was added and the reaction mixture was sparged with nitrogen atmosphere. The suspension from the first vial was transferred to a syringe. Then a syringe filter and new needle were installed on the syringe, and 1 mL of the solution was injected through the syringe filter into the DMAc solution. The resulting reaction mixture was sparged with nitrogen for 15 min. The vial was irradiated with 2 blue LEDs Photoreaction at room temperature for 16 hours. LCMS showed that the desired product was formed. The reaction mixture was quenched by addition of saturated NH4CI (20 mL). The aqueous layer was extracted with EtOAc (3 x 150 mL). The combined organic phase was washed with brine (70 mL), dried over anhydrous Na2SC>4. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (5:1) to afford tert-butyl (4-(3,3-difluorocyclobutyl)-3-fluorophenyl) carbamate (160 mg, 50.02% yield, 97% purity) as a white solid. LC / MS: MS (ESI) calcd for C15H18F3NO2: 301.13. Found: 302.15[M+H]+.
[2221] Step 6. Synthesis of 4-(3,3-difluorocyclobutyl)-3-fluoroanilines
[2222]
[2223] To a stirred solution of tert-butyl (4-(3,3-difluorocyclobutyl)-3-fluorophenyl) carbamate (120 mg, 0.398 mmol, 1 equiv) in DCM (4 mL) was added TFA (1 mL) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h under nitrogen atmosphere. After completion of the reaction. The resulting mixture was diluted with DCM (10 mL) and quenched with saturated NaHCOs (aq.) at 0°C. The aqueous layer was extracted with DCM (10 mL x 3). The combined organic phase was washed with sodium bicarbonate aqueous solution, dried over anhydrous sodium sulfate and concentrated under reduced pressure. This resulted in 4-(3,3-difluorocyclobutyl)-3-fluoroanilines (80 mg, crude product) as a black solid. LC / MS: MS (ESI) calcd for C10H10F3N: 201.08. Found: 202.15[M+H]+.PATENT
[2224] ATTORNEY-DOCKET NO.: 51478-036WO4
[2225] Step 7. Synthesis of (R)-8-((4-(3,3-difluorocyclobutyl)-3-fluorophenyl) amino)-3-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2226] " S ' '" 'C -l NHS" Yi 'NH
[2227] DIEA. DMSO %
[2228] Y,. J, £ &
[2229] r' N N 'ff '• 70‘G, 5 hr f'’ N' N y
[2230] G O
[2231]
[2232] To a stirred solution of 3-methyl-6-((R)-2-methylmorpholino)-8-(methylsulfinyl) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 0.186 mmol, 1 equiv) and 4-(3,3-difluorocyclobutyl)-3-fluoroaniline (44.80 mg, 0.223 mmol, 1.2 equiv) in DMSO (5 mL) was added DIEA (47.96 mg, 0.372 mmol, 2 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 50°C for 1h under nitrogen atmosphere. LCMS showed that the desired product was formed. The reaction mixture was quenched by addition of saturated NH4CI (20 mL). The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in (R)-8-((4-(3,3-difluorocyclobutyl)-3-fluorophenyl) amino)-3-methyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one as a trifluoroacetate salt (15.5 mg, 14.54% yield, 99.5% purity) and as a yellow solid. LC / MS: MS (ESI) calcd for C22H23F3N6O2: 460.18. Found: 461.10[M+H]+ 1H NMR (300 MHz, DMSO-d6) 6: 8.27 (s, 1 H), 7.81 - 7.86 (m, 1 H), 7.68 - 7.78 (m, 1 H), 7.37 - 7.43 (m, 1 H), 4.44 - 4.51 (m, 2H), 3.93 - 3.95 (m, 1 H), 3.51 (s, 6H), 2.68 - 3.18 (m, 3H), 2.76 - 2.89 (m, 3H), 1.17 - 1.23 (m, 3H).
[2233] Using the approach to synthesize Compound 122, the compounds of Table 9 were synthesized from morpholine and the corresponding aniline.
[2234] Table 9: Experimental Data for Compounds
[2235] Ex Cmpd
[2236] Name Structure MS 1H NMR
[2237] # #
[2238] 119 123 (R)-8-((3-fluoro-4- 439.05 1H NMR (300 MHz, DMSO) 6 (trifluoromethyl)pheny [M+H]+10.18 (s, 1H), 8.25 - 8.27 (m, F
[2239] l)amino)-3-methyl-6- v yk 2H), 7.98 - 8.01 (m, 1H), 7.72 - (2- 7.78 (m, 1H), 4.43 - 4.49 (m, methylmorpholino)pyr 2H), 3.91 - 3.96 (m, 1H), 3.50 - f ’N” v
[2240] imido[5,4-d]pyrimidin- kA 6 3.55 (m, 5H), 2.98 - 3.07 (m, 4(3H)-one 1H), 2.70 - 2.74 (m, 1H), 1.17 - 1.19 (m, 3H).
[2241]
[2242] PATENT
[2243] ATTORNEY-DOCKET NO.: 51478-036WO4
[2244] 120 124 8-((3-fluoro-4- 425.05 1H NMR (300 MHz, DMSO) 6 F
[2245] (trifluoromethyl)pheny FSCX[M+H]+10.18 (s, 1H), 8.26 (s, 1H), 8.03 - l)amino)-3-methyl-6- 8.13 (m, 2H), 7.73 - 7.79 (m, morpholinopyrimido[5 ■A-A. 1H), 3.69 - 3.76 (m, 8H), 3.50 - 1 1
[2246] ,4-d]pyrimidin-4(3H)- f M Y
[2247] Y. J 0 3.52 (m, 3H).
[2248] one
[2249] 121 125 (R)-8-((4- 450.20 1H NMR (400 MHz, DMSO-d6) 6 isopropylphenyl)amin [M+H]+(ppm): 7.94 (s, 1H), 7.74 - 7.76 o)-3-(1- (m, 2H), 7.26 - 7.28 (m, 2H), methylazetidin-3-yl)- 5.07 - 5.15 (m, 1H), 4.81 - 4.83 6-(2- (m, 1H), 4.60 - 4.67 (m, 2H), methylmorpholino)pyr....-U JI. 4.51 - 4.54 (m, 1H), 4.43 - 4.46 imido[5,4-d]pyrimidin- YNAA
[2250] a J 0 (m, 2H), 3.92 - 3.95 (m, 1H), 4(3H)-one I 3.50 - 3.53 (m, 2H), 3.01 - 3.04 (m, 2H), 2.90 - 2.94 (m, 2H), 2.87 - 2.89 (m, 1H), 2.68 - 2.72 (m, 1H), 1.89 - 1.22 (m, 9H).
[2251]
[2252] Example 122: (R)-8-((4-(3,3-difluorocyclobutyl)-3-fluorophenyl)amino)-3-methyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 126)
[2253] Step 1. Synthesis of 5-amino-2-chloro-N-cyclopropyl-6-methoxypyrimidine-4-carboxamide
[2254] HATH, DIEA. DMFfctX. NH>
[2255] Q" N f hr ' N y '-77
[2256]
[2257] O O
[2258] To a solution of 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid (2 g, 9.824 mmol, 1 equiv) and aminocyclopropane (0.67 g, 11.789 mmol, 1.2 equiv) in DMF (20 mL) were added HATU (4.48 g, 11.789 mmol, 1.2 equiv) and DIEA (3.81 g, 29.472 mmol, 3 equiv) at room temperature. The final reaction mixture was irradiated with microwave radiation at room temperature for 1 h. The reaction was monitored by LCMS. LCMS showed the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 1% to 70% gradient in 20 min; detector, UV 254 nm. This resulted in 5-amino-2-chloro-N-cyclopropyl-6-methoxypyrimidine-4-carboxamide (2 g, 83.90% yield, 96% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C9H11CIN4O2: 242.66, Found: 243.05[M+H]+.PATENT
[2259] ATTORNEY-DOCKET NO.: 51478-036WO4
[2260] Step 2. Synthesis of 6-chloro-3-cyclopropyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one
[2261] I
[2262] Y < " O
[2263] ... X -W MOAc 1,. N.
[2264] A --A N A N.
[2265] GT M" Y '" V7 120'C. Cl' N" Y -V7
[2266]
[2267] O7O
[2268] To a solution of 5-amino-2-chloro-N-cyclopropyl-6-methoxypyrimidine-4-carboxamide (2.4 g, 9.890 mmol, 1 equiv) in trimethoxymethane (25 mL) was added acetic acid (1.78 g, 29.670 mmol, 3 equiv) at room temperature. The final reaction mixture was irradiated with microwave radiation at 120°C for 1h. The reaction was monitored by LCMS. LCMS showed the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 1% to 70% gradient in 20 min; detector, UV 254 nm. This resulted in 6-chloro-3-cyclopropyl-8-methoxypyrimido[5,4-d] pyrimidin-4(3H)-one (480 mg, 19.21% yield, 98% purity) as a white solid.
[2269] LC / MS: MS (ESI) calcd. for C10H9CIN4O2: 252.66. Found: 253.15[M+H]+.
[2270] Step 3. Synthesis of (R)-3-cyclopropyl-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2271] Y'NM
[2272] 6. J
[2273] -1
[2274] D1EA. DMSO 'N.
[2275] . A; A, xN, - T J
[2276] Ci HIf ‘V W0'C, 2 hre " V?
[2277] 0&
[2278]
[2279] To a solution of 6-chloro-3-cyclopropyl-8-methoxypyrimido[5,4-d] [1,3] diazin-4-one (480 mg, 1.900 mmol, 1 equiv) and (2R)-2-methylmorpholine (230.6 mg, 2.280 mmol, 1.2 equiv) in DMSO (6 mL) was added DIEA (736.6 mg, 5.700 mmol, 3 equiv) at room temperature. The final reaction mixture was irradiated with microwave radiation at 100°C for 2 h. The reaction was monitored by LCMS. LCMS showed the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 5% to 90% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-3-cyclopropyl-8-methoxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (600 mg, 99.52% yield, 98% purity) as a light yellow solid. LC / MS: MS (ESI) calcd. for C15H19N5O3:
[2280] 317.35. Found: 318.05[M+H]+.PATENT
[2281] ATTORNEY-DOCKET NO.: 51478-036WO4
[2282] Step 4. Synthesis of (R)-3-cyclopropyl-6-(2-methylmorpholino)-8-((4-(trifluoromethyl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2283] KHMD8, THF f # N if X7 80’C, 1 hr r* T8 NY 'Y~?
[2284] ov v
[2285]
[2286] To a solution of (R)-3-cyclopropyl-8-methoxy-6-(2-methylmorpholino)pyrimido[5,4-d] pyrimidin-4(3H)-one (550 mg, 1.733 mmol, 1 equiv) and P-trifluoromethylaniline (418.9 mg, 2.600 mmol, 1.50 equiv) in THF (22 mL) was added KHMDS (1.0M in THF) (2.6 mL, 1.5 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred at 0°C for additional 6 min. The final reaction mixture was irradiated with microwave radiation at 80°C for 1h. The reaction was monitored by LCMS. LCMS showed the reaction was completed, the resulting mixture was concentrated under reduced pressure. The reaction was quenched with sat. NH₄Cl (aq)(100 mL) at 0°C. The resulting mixture was extracted with EtOAc (2 x 300 mL). The combined organic layers were washed with brine (1 x 100 mL) The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 5% to 95% gradient in 20 min; detector, UV 254 nm. This resulted in (R)-3-cyclopropyl-6-(2-methylmorpholino)-8-((4-(trifluoromethyl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (317 mg, 40.97% yield, 98.0% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C21H21F3N6O2: 446.43. Found: 447.05[M+H]+.1H NMR (400 MHz, DMSO-d₆) 6 (ppm): 9.91 (s, 1 H), 8.12 - 8.22 (m, 3H), 7.72 - 7.74 (m, 2H), 4.44 - 4.51 (m, 2H), 3.91 - 3.94 (m, 1H), 3.47 - 3.55 (m, 2H), 3.23 - 3.26 (m, 1H), 2.98 - 3.04 (m, 1H), 2.65 - 2.71 (m, 1H), 1.17 - 1.19 (m, 3H), 1.04 - 1.09 (m, 2H), 0.92 - 0.96 (m, 2H).
[2287] Using the approach to synthesize Compound 126, the compounds of Table 10 were synthesized from morpholine and the corresponding aniline.
[2288] Table 10: Experimental Data for Compounds
[2289] Ex Cmpd
[2290] Name Structure MS 1H NMR
[2291] # #
[2292] 123 127 (S)-3-ethyl-6-(3- 435.10 1H NMR (400 MHz, DMSO-d6) 6 methylmorpholino)-8- [M+H]+(ppm): 9.95 - 10.04 (m, 1H), 8.30 ((4-(trifluoromethyl)- - 8.35 (m, 1H), 8.10 - 8.20 (m, phenyl)amino)pyrimid 2H), 7.75 - 7.85 (m, 2H), 4.65 - o[5,4-d]pyrimidin- 7.78 (m, 1H), 4.28 - 4.35 (m, 4(3H)-one r N w Y N x 1H), 4.00 - 4.10 (m, 2H), 3.90 - 4.00 (m, 1H), 3.70 - 3.80 (m, 1H), 3.55 - 3.65 (m, 1H), 3.40 - 3.50 (m, 1H), 3.20 - 3.30(m, 1H), 1.20 - 1.40(m, 6H).
[2293]
[2294] PATENT
[2295] ATTORNEY-DOCKET NO.: 51478-036WO4
[2296] 124 128 (S)-6-(3- 491.15 1H NMR (400 MHz, DMSO-d6) 6 methylmorpholino)-3- [M+H]+(ppm): 8.27 (s, 1H), 8.13 - 8.16 (tetrahydro-2H-pyran- (m, 2H), 7.72 - 7.74 (m, 2H), 4-yl)-8-((4- 4.81 - 4.85 (m, 1H), 4.65 - 4.67 (trifluoromethyl)pheny X,„ (m, 1H), 4.27 - 4.31 (m, 1H), l)amino)pyrimido[5,4- A XS 4.00 - 4.04 (m, 2H), 3.94 - 3.97
[2297] N._,
[2298] d]pyrimidin-4(3H)-one (m, 1H), 3.73 - 3.76 (m, 2H),
[2299] 3.58 - 3.61 (m, 2H), 3.57 - 3.52 (m, 1H), 3.18 - 3.25 (m, 1H), 2.19 - 2.21 (m, 2H), 1.86 - 1.89 (m, 2H),1.19 - 1.21 (m, 3H).
[2300]
[2301] Example 125: (R)-8-((4-isopropylphenyl)amino)-3-(2-methoxyethyl)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 129)
[2302] Step 1. Synthesis of methyl 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylate
[2303] a ’""s
[2304] 4, X„ NH- CH3SNa, THF,, X. NH
[2305] N -'-r N:<<;i
[2306] X,0.4. A..0.
[2307]
[2308] C: N Y 1 hr cr N 'IT
[2309] O O
[2310] To a solution of methyl 5-amino-2,6-dichloropyrimidine-4-carboxylate (10 g, 45.039 mmol, 1 equiv) in anhydrous THF (100 mL) was added CH₃SNa (3.47 g, 49.543 mmol, 1.1 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for a period of 1 h. Desired product could be detected by LCMS. After completion of the reaction, the reaction mixture was quenched by addition of saturated NH4CI (100 mL) at 0 °C. The aqueous layer was extracted with ethyl acetate (3 x 100 mL). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford methyl 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylate (7.9 g, 75.06% yield, 92% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C7H8CIN3O2S: 233.67, Found: 234.10[M+H]+.
[2311] Step 2. Synthesis of 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylic acid
[2312] LiOH, THF, MeOH
[2313] CT N -|f - rt, W
[2314]
[2315] O
[2316] To a solution of methyl 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylate (7.9 g, 33.808 mmol, 1 equiv) in THF (50 mL), MeOH (25 mL) and H2O (50 mL) was added LiOH (1.62 g, 67.616 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at room temperature for a period of 1 h. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was acidified to pH = 5 with 2 M HCI (aq.). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product, which wasPATENT
[2317] ATTORNEY-DOCKET NO.: 51478-036WO4
[2318] further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylic acid (5 g, 67.33% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for C6H6CIN3O2S: 218.99, Found: 220.05[M+H]+.
[2319] Step 3. Synthesis of 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxamide
[2320] ■' S '" S
[2321] X UM. NH.. OS, HATU. D1EA. DMF X. NH,
[2322] Ft. i hr VNHie
[2323]
[2324] 6 6
[2325] To a solution of 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylic acid (5 g, 22.765 mmol, 1 equiv) in anhydrous DMF (50 mL) were added NH4CI (1.84 g, 34.148 mmol, 1.5 equiv), HATU (12.9 g, 34.148 mmol, 1.5 equiv) and DIEA (5.8 g, 45.530 mmol, 2 equiv) at 0 °C. The reaction mixture was stirred at 25 °C for a period of 1 h. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (100 mL). The aqueous layer was extracted with ethyl acetate (3 x 100 mL). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxamide (7.9 g, 75.06% yield, 92% purity) as a white solid. LC / MS: MS (ESI) calcd. for C6H7CIN4OS: 218.00, Found: 219.15[M+H]+.
[2326] Step 4. Synthesis of 6-chloro-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2327] I
[2328] HOAc A.
[2329] ■yN^ 120’C, 16 hrs
[2330]
[2331] a 6
[2332] To a solution of 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxamide (3.5 g, 16.007 mmol, 1 equiv) in anhydrous trimethoxymethane (30 mL) was added AcOH (2.88 g, 48.021 mmol, 3 equiv) at room temperature. The reaction mixture was stirred at 120 °C for a period of 16 h. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (50 mL). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford 6-chloro-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (3 g, 81.97% yield, 90% purity) as a white solid. LC / MS: MS (ESI) calcd. for C7H5CIN4OS: 227.99, Found: 229.10[M+H]+.PATENT
[2333] ATTORNEY-DOCKET NO.: 51478-036WO4
[2334] Step 5. Synthesis of (R)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2335] QfEA. DMSO
[2336] I'-"' N
[2337] O. G
[2338]
[2339] To a solution of 6-chloro-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (350 mg, 1.531 mmol, 1 equiv) in anhydrous DMSO (5 mL) were added (R)-2-methylmorpholine (232.2 mg, 2.296 mmol, 1.5 equiv) and DIEA (395.7 mg, 3.062 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 100 °C for a period of 1 h. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL) at 0 °C. The aqueous layer was extracted with ethyl acetate (3 x 80 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford (R)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (320 mg, 71.26% yield, 95% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C12H15N5O2S: 293.09, Found: 294.10[M+H]+.
[2340] Step 6. Synthesis of (R)-3-(2-methoxyethyl)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2341] D. O C. 1 hr o j o
[2342]
[2343] To a stirred solution of (R)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (320 mg, 1.091 mmol, 1 equiv) in anhydrous DMF (5 mL) were added 2-bromoethyl methyl ether (181.9 mg, 1.309 mmol, 1.2 equiv) and Cs₂CO₃ (710.8 mg, 2.182 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 70 °C for a period of 1 h. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL). The aqueous layer was extracted with ethyl acetate (3 x 60 mL). The combined organic phase was washed with brine (25 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford (R)-3-(2-methoxyethyl)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (300 mg, 78.26% yield, 95% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H21N5O3S: 351.14, Found: 352.15[M+H]+.PATENT
[2344] ATTORNEY-DOCKET NO.: 51478-036WO4
[2345] Step 7. Synthesis of (R)-3-(2-methoxyethyl)-6-(2-methylmorpholino)-8-(methylsulfonyl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2346] Q
[2347] 's X. A
[2348] N X' nfrCPBA, DGMrS '!
[2349] • I.4 w X z'A M
[2350] XX' - *• <" Y ’N' Y °
[2351] 6: 6 25 c, 1 hr 6 J o
[2352]
[2353] T T
[2354] To a solution of (R)-3-(2-methoxyethyl)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (280 mg, 0.830 mmol, 1 equiv) in anhydrous DCM (5 mL) was added m-CPBA (157.5 mg, 0.913 mmol, 1.1 equiv) at room temperature. The reaction mixture was stirred at 25 °C for a period of 1 h. After completion of reaction, the reaction mixture was quenched by addition of saturated Na₂S₂O₃ (25 mL). The aqueous layer was extracted with ethyl acetate (3 x 40 mL). The combined organic phase was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford (R)-3-(2-methoxyethyl)-6-(2-methylmorpholino)-8-(methylsulfonyl) pyrimido[5,4-d] pyrimidin-4(3H)-one (220 mg, 71.77% yield, 90% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H21N5O5S: 383.13, Found: 384.10[M+H]+.
[2355] Step 8. Synthesis of (R)-8-hydroxy-3-(2-methoxyethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2356] p
[2357] OH
[2358] M-XA DIEA, EMWSO, X N
[2359] 1 1 1 »• v
[2360] f" -N- -hr y were, 1 hr XNAN*'YN'' / V
[2361] X. X o X J o
[2362]
[2363] To a solution of (R)-3-(2-methoxyethyl)-6-(2-methylmorpholino)-8-(methylsulfonyl) pyrimido[5,4-d] pyrimidin-4(3H)-one (200 mg, 0.522 mmol, 1 equiv) in DMSO (5 mL) and H2O (0.1 mL) was added DIEA (134.8 mg, 1.044 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 100 °C for a period of 1 h. After completion of reaction, the reaction mixture was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford (R)-8-hydroxy-3-(2-methoxyethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (140 mg, 83.52% yield, 92% purity) as a white solid. LC / MS: MS (ESI) calcd. for C14H19N5O4: 321.14, Found: 322.20[M+H]+.PATENT
[2364] ATTORNEY-DOCKET NO.: 51478-036WO4
[2365] Step 9. Synthesis of (R)-8-chloro-3-(2-methoxyethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2366] C:
[2367] POCl3 -...-k. A.,
[2368] jl J I _ - ». 'i 1 r 'ox«rc,1 hr
[2369] 0■ ° c J 6
[2370]
[2371] t 1
[2372] A solution of (R)-8-hydroxy-3-(2-methoxyethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (120 mg, 0.373 mmol, 1 equiv) in anhydrous POCl3 (2 mL) was stirred at 80 °C for a period of 1 h. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was concentrated under reduced pressure. The residue was basified to pH 7-8 with saturated NaHCCh (aq.) at 0°C. The aqueous layer was extracted with ethyl acetate (3 x 25 mL). The combined organic phase was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give (R)-8-chloro-3-(2-methoxyethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (80 mg, 63.05% yield, 94% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C14H18CIN5O3: 339.11, Found: 340.20[M+H]+.
[2373] Step 10. Synthesis of (R)-8-((4-isopropylphenyl)amino)-3-(2-methoxyethyl)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[2374] A ftd ’ *■ W''V DIEA.& MSO %
[2375] i-'‘ N' «• y ' a wc, i hrv-o"'
[2376] ;O f>.. J 6
[2377]
[2378] To a solution of (R)-8-chloro-3-(2-methoxyethyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 0.177 mmol, 1 equiv) in anhydrous DMSO (2 mL) were added paraisopropylaniline (28.7 mg, 0.212 mmol, 1.2 equiv) and DIEA (45.7 mg, 0.354 mmol, 2 equiv) at room temperature. The reaction mixture was stirred 100 °C for a period of 1 h. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (25 mL). The aqueous layer was extracted with ethyl acetate (3 x 25 mL). The combined organic phase was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford (R)-8-((4-isopropylphenyl)amino)-3-(2-methoxyethyl)-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one; trifluoroacetic acid salt (35.4 mg, 36.28% yield, 99.354% purity) as a light yellow solid. LC / MS: MS (ESI) calcd. for C23H30N6O3: 438.53, Found: 439.20[M+H]+.1H NMR (400 MHz, DMSO-d6) 6 (ppm): 9.62 - 9.70 (m, 1H), 8.16 - 8.17 (m, 1H), 7.77 - 7.80 (m, 2H), 7.23 - 7.25 (m, 2H), 4.40 - 4.58 (m, 2H), 4.15 - 4.18 (m, 2H), 3.90 - 3.94 (m, 1H), 3.52 - 3.62 (m, 2H), 3.46 - 3.49 (m, 2H), 3.25 - 3.26 (m, 3H), 2.97 - 3.04 (m, 1H), 2.84 - 2.91 (m, 1H), 2.65 - 2.70 (m, 1H), 1.15 - 1.22 (m, 9H).PATENT
[2379] ATTORNEY-DOCKET NO.: 51478-036WO4
[2380] Example 126: (R)-8-((4-(2,2-difluoroethyl)phenyl)amino)-3-ethyl-6-(2-methylmorpholino)pyrimido-[5,4-d]pyrimidin-4(3H)-one (Compound 130)
[2381] Step 1. Synthesis of tert-butyl (4-(2,2-difluoroethyl)phenyl)carbamate
[2382] p.. ~ NHjBac. Ephss RS G4, Ephos, Cs-Cos. dstsxane
[2383] I I: -sJ I -:
[2384] F < F < x X 'Br 80 ’C. 1 hr 'N' ‘O’
[2385]
[2386] K
[2387] To a solution of 1-bromo-4-(2,2-difluoroethyl)benzene (1 g, 4.524 mmol) in 13 ml of dioxane were added NH2B0C (0.79 g, 6.786 mmol), Ephos Pd G4 (0.42 g, 0.452 mmol), Cs₂CO₃ (3.68 g, 11.310 mmol) and Ephos (0.48 g, 0.905 mmol) at room temperature. The resulting mixture was stirred at 80 °C for 1 h under N2. After completion of reaction and cooled to room temperature, the reaction mixture was further purified by reverse phase column eluted with ACN and H2O (0.05% NH4HCO3), after concentrated to afford tert-butyl (4-(2,2-difluoroethyl)phenyl)carbamate (200 mg, 17.18% yield) as a white solid. MS: MS (ESI) calcd. for C13H17F2NO2: 257.28 Found: 258.05[M+H]+.
[2388] Step 2. Synthesis of 4-(2,2-difluoroethyl)aniline
[2389] DCM HCLdioxane
[2390] .
[2391]
[2392] 0 C - rt 0.5 hr
[2393] To a solution of tert-butyl (4-(2,2-difluoroethyl)phenyl)carbamate (60 mg, 0.233 mmol) in 0.9 ml of DCM was added 0.3 ml of HCI in dioxane (4M) at 0 °C. The resulting mixture was stirred at room temperature for 0.5 h. After completion of reaction, the reaction mixture was concentrated under reduced pressure to afford 4-(2,2-difluoroethyl)aniline (52.9 mg, crude) as a white solid. LC / MS: MS (ESI) calcd. for C8H9F2N: 157.16 Found: 158.00[M+H]+.
[2394] Step 3. Synthesis of (R)-8-((4-(2,2-difluoroethyl)phenyl)amino)-3-ethyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[2395] r: IFA. DMSQ, 50' C - 100 X,
[2396]
[2397] To a solution of 4-(2,2-difluoroethyl)aniline (52.91 mg, 0.337 mmol) in 2 ml of DMSO were added 6,8-dichloro-3-ethylpyrimido[5,4-d]pyrimidin-4(3H)-one (75 mg, 0.306 mmol) and DIEA (197.78 mg, 1.530 mmol) at room temperature. The resulting mixture was stirred at 50 °C for 1 h. After cooled to room temperature. Then, to the above mixture was added (R)-2-methylmorpholine (46.43 mg, 0.459 mmol) at room temperature and stirred at 100 °C for 1 h. After completion of reaction and cooled to room temperature, the reaction mixture was further purified by reverse phase column eluted with ACN and H2O (0.05% TFA), after concentrated to afford (R)-8-((4-(2,2-difluoroethyl)phenyl)amino)-3-ethyl-6-(2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (57.7 mg, 34.63% yield) as a trifluoroacetate and as a yellow solid. LC / MS: MS (ESI) calcd. for C21H24F2N6O2: 430.46 Found: 431.10[M+H]+.1H NMR (400 MHz, DMSO-de) 6 (ppm): 9.99 (s, 1H), 8.31 (s, 1 H), 7.73 - 7.86 (m, 2H), 7.21 - 7.40(m, 2H), 6.02 -PATENT
[2398] ATTORNEY-DOCKET NO.: 51478-036WO4
[2399] 6.41 (m, 1 H), 4.28 - 4.51 (m, 2H), 3.98 - 4.11 (m, 2H), 3.89 - 3.97 (m, 1 H), 3.39 - 3.59 (m, 2H), 3.11 - 3.25 (m, 2H), 2.91 - 3.10 (m, 1H), 2.60 - 2.75 (m, 1H), 1.26 - 1.37 (m, 3 H), 1.14 - 1.20 (m, 3H).
[2400] Example 127: (R)-6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one and (S)-6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 131 and Compound 132)
[2401] Step 1. Synthesis of 6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl) phenyl) amino)-3-methylpyrimido[5,4-d] pyrimidin-4(3H)-one
[2402]
[2403] 2. D£A. DMSO, 10OCC, Ihf
[2404] To a stirred solution of 6,8-dichloro-3-methylpyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.433 mmol, 1 equiv) and 4-(difluoromethyl) aniline (74.34 mg, 0.520 mmol, 1.2 equiv) in DMSO (4 mL) was added DIEA (111.88 mg, 0.866 mmol, 2 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 50°C for 1h under nitrogen atmosphere. LCMS showed the reaction was completed. The solution was cooled to room temperature. To above solution was added 2-cyclopropylmorpholine (45.19 mg, 0.355 mmol, 1.2 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100°C for 1h under nitrogen atmosphere. After completion of reaction monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in 6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl) phenyl) amino)- 3-methylpyrimido[5,4-d] pyrimidin-4(3H)-one (90 mg, 70.94% yield, 96% purity) as a white solid. LC / MS: MS (ESI) calcd for C21H22F2N6O2: 428.18. Found: 429.10[M+H]+.
[2405] Step 2. Separation of (R)-6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one and 3166 (S)-6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one
[2406]
[2407] The product (90 mg) was purified by silica gel Column: (R, R)-WHELK-O1-Kromasil, 5 pm, 250 mm x 20 mm; Mobile Phase A: Hex (with 0.5% NH3 (2 M in MeOH)), Mobile Phase B: (1: 1 EtOH: DCM);PATENT
[2408] ATTORNEY-DOCKET NO.: 51478-036WO4
[2409] Flow rate: 20 mL / min; Gradient (B%)40% B; Wave Length: 220 / 254 nm; Sample Solvent: EtOH+DCM; Injection Volume: 0.5 mL; Number Of Runs: 20; Single injection run time(min): 18 to afford
[2410] The first eluting compound, (R)-6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one (29.2 mg, 32.44% yield, 98.7% purity) as a white solid. Stereochemistry arbitrarily assigned.
[2411] LC / MS: MS (ESI) calcd for C21H22F2N6O2: 428.18. Found: 429.10[M+H]+
[2412] 1H NMR (400 MHz, DMSO) 6: 9.84 (s, 1H), 8.24 (s, 1H), 8.04 - 8.06 (m, 2H), 7.59 - 7.52 (m, 2H), 6.87 - 7.15 (m, 1 H), 4.38 - 4.58 (m, 2H), 3.90 - 3.98 (m, 1H), 3.50 (s, 3H), 3.38 - 3.48 (m, 1H), 3.07 - 3.33 (m, 1 H), 2.83 - 2.93 (m, 1 H), 2.74 - 2.77 (m, 1 H), 0.81 - 0.99 (m, 1 H), 0.51 - 0.54 (m, 2H), 0.24 - 0.39 (m, 2H).
[2413] The second eluting compound (S)-6-(2-cyclopropylmorpholino)-8-((4-(difluoromethyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one (28.5 mg, 31.67% yield, 99.0% purity) as a white solid. Stereochemistry arbitrarily assigned.
[2414] LC / MS: MS (ESI) calcd for C21H22F2N6O2: 428.18. Found: 429.10[M+H]+
[2415] 1H NMR (400 MHz, DMSO) 6: 9.84 (s, 1H), 8.24 (s, 1H), 8.04 - 8.06 (m, 2H), 7.59 - 7.52 (m, 2H), 6.87 - 7.15 (m, 1H), 4.55 - 4.58 (m, 1H), 4.38 - 4.55 (m, 1H), 3.90 - 3.98 (m, 1H), 3.50 (s, 3H), 3.38 - 3.48 (m, 1 H), 3.07 - 3.33 (m, 1 H), 2.83 - 2.93 (m, 1 H), 2.74 - 2.77 (m, 1 H), 0.81 - 0.99 (m, 1 H), 0.51 -0.54 (m, 2H), 0.24 - 0.39 (m, 2H).
[2416] Example 128: 3-ethyl-6-((R)-2-methylmorpholino)-8-((4-((R)-tetrahydrofuran-2-yl)phenyl)amino)-pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-ethyl-6-((R)-2-methylmorpholino)-8-((4-((S)-tetrahydro-furan-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 133 and Compound 134) Step 1. Synthesis of tert-butyl (4-(tetrahydrofuran-2-yl) phenyl) carbamate
[2417] g?, NIBr.tfyme. fflbbpy. KJ-SPO,,. RsCHO, THF < \
[2418] ¥ i ~ <? t,
[2419] N ' o'^ rt. 72 hrs N O
[2420]
[2421] H H
[2422] To a solution of tert-butyl (4-bromophenyl) carbamate (1 g, 3.675 mmol, 1 equiv) in anhydrous THF (10 mL, 123.428 mmol) were added NiBr₂·glyme (113.4 mg, 0.367 mmol, 0.1 equiv), dtbbpy (98.6 mg, 0.367 mmol, 0.1 equiv), PhCHO (194.9 mg, 1.837 mmol, 0.5 equiv) and K2HPO4 (1.28 g, 7.350 mmol, 2 equiv) at room temperature under N2 atmosphere. The reaction mixture was stirred at room temperature for a period of 72 h irradiated with blue LED under N2 atmosphere. Desired product could be detected by LCMS. After completion of reaction, the reaction mixture was quenched by addition of saturated NH4CI (50 mL) at 0 °C. The aqueous layer was extracted with ethyl acetate (3 x 200 mL). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product, which was further purified by silica gel column chromatography, eluted with PE / EA (3:1) to afford tert-butyl (4-(tetrahydrofuran-2-yl) phenyl) carbamate (240 mg, 24.80% yield, 94% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H21NO3: 263.15, Found: 262.10[M-H]+.PATENT
[2423] ATTORNEY-DOCKET NO.: 51478-036WO4
[2424] Step 2. Synthesis of 4-(tetrahydrofuran-2-yl) aniline
[2425] ■( X...., ( 1
[2426] ° T *1 f L' TFA DCM
[2427]
[2428] To a solution of tert-butyl (4-(tetrahydrofuran-2-yl) phenyl) carbamate (240 mg, 0.911 mmol, 1 equiv) in anhydrous DCM (3 mL) was added TFA (1 mL) at room temperature. The reaction mixture was stirred at room temperature for a period of 1 h. Desired product could be detected by LCMS. After completion of the reaction, the reaction mixture was basified to pH = 8 with saturated NaHCOs (aq.). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was washed with brine (30 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by silica gel column chromatography, eluted with PE / EA (1:2) to afford 4-(tetrahydrofuran-2-yl) aniline (90 mg, 60.50% yield, 93% purity) as a light yellow solid. LC / MS: MS (ESI) calcd. for C10H13NO: 163.10, Found: 164.15[M+H]+.
[2429] Step 3. Synthesis of 3-ethyl-6-((R)-2-methylmorpholino)-8-((4-(tetrahydrofuran-2-yl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2430]
[2431] To a solution of (R)-8-chloro-3-ethyl-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (113.8 mg, 0.367 mmol, 1 equiv) in anhydrous DMSO (2 mL) were added 4-(tetrahydrofuran-2-yl) aniline (90 mg, 0.551 mmol, 1.5 equiv) and DIEA (95.0 mg, 0.735 mmol, 2 equiv) at room temperature. The reaction mixture was stirred at 100 °C for a period of 2 h. Desired product could be detected by LCMS. After completion of the reaction, the reaction mixture was quenched by the addition of saturated NH4CI (30 mL) at 0 °C. The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give crude product which was further purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.05% TFA), 10% to 70% gradient in 20 min; detector, UV 254 nm to afford 3-ethyl-6-((R)-2-methylmorpholino)-8-((4-(tetrahydrofuran-2-yl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (90 mg, 56.09% yield, 95% purity) as a light yellow solid. LC / MS: MS (ESI) calcd. for C23H28N6O3: 436.52, Found: 437.10[M+H]+.PATENT
[2432] ATTORNEY-DOCKET NO.: 51478-036WO4
[2433] Step 4. Separation of 3-ethyl-6-((R)-2-methylmorpholino)-8-((4-((R)-tetrahydrofuran-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-ethyl-6-((R)-2-methylmorpholino)-8-((4-((S)-tetrahydrofuran-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[2434]
[2435] The product (90 mg) was separated by SFC flash chromatography with the following conditions: Column: CHIRALPAK IB, 5 pm, 250 mm x 20 mm; Mobile Phase A: Hex (with 0.5% NH3 (2 M in MeOH)), Mobile Phase B: EtOH: DCM=1: 1; Flow rate: 20 mL / min; Gradient (B%)10% B; Wave Length: 220 / 254 nm; Sample Solvent: EtOH--HPLC; Injection Volume: 0.5 mL; Number Of Runs: 10; Single injection run time(min): 55.
[2436] to afford 3-ethyl-6-((R)-2-methylmorpholino)-8-((4-((R)-tetrahydrofuran-2-yl)phenyl)amino)pyrimido[5,4-d]pyrimidin-4(3H)-one (22.0 mg, 24.44% yield, 95.021% purity) as a light yellow solid. Stereochemistry arbitrarily assigned.
[2437] RT1(min): 41.45
[2438] LC / MS: MS (ESI) calcd. for C23H28N6O3: 436.52, Found: 437.10[M+H]+.
[2439] 1H NMR (300 MHz, DMSO-d6) 6 (ppm): 9.54 - 9.55 (m, 1H), 8.25 - 8.26 (m, 1H), 7.82 - 7.83 (m, 2H), 7.28 - 7.29 (m, 2H), 4.76 - 4.80 (m, 1H), 4.40 - 4.53 (m, 2H), 3.90 - 4.04 (m, 4H), 3.75 - 3.82 (m, 1H), 3.45 - 3.52 (m, 2H), 2.94 - 3.03 (m, 1 H), 2.61 - 2.69 (m, 1 H), 2.24 - 2.30 (m, 1 H), 1.96 - 1.99 (m, 2H), 1.62 - 1.71 (m, 1H), 1.22 - 1.28 (m, 3H), 1.16 - 1.20 (m, 3H).
[2440] And also to obtain 3-methyl-6-((R)-2-methylmorpholino)-8-((4-((S)-tetrahydrofuran-2-yl) phenyl) amino) pyrimido[5,4-d] pyrimidin-4(3H)-one (15.0 mg, 16.6% yield, 96.823% purity) a light yellow solid. Stereochemistry arbitrarily assigned.
[2441] RT2(min): 48.45
[2442] LC / MS: MS (ESI) calcd. for C23H28N6O3: 436.52, Found: 437.15[M+H]+.
[2443] 1H NMR (300 MHz, DMSO-d6) 6 (ppm): 9.56 - 9.56 (m, 1H), 8.25 - 8.26 (m, 1H), 7.82 - 7.86 (m, 2H), 7.28 - 7.32 (m, 2H), 4.74 - 4.80 (m, 1H), 4.40 - 4.53 (m, 2H), 3.89 - 4.04 (m, 4H), 3.72 - 3.83 (m, 1H), 3.40 - 3.52 (m, 2H), 2.94 - 3.02 (m, 1 H), 2.62 - 2.69 (m, 1 H), 2.22 - 2.30 (m, 1 H), 1.94 - 1.97 (m, 2H), 1.61 - 1.73 (m, 1H), 1.23 - 1.30 (m, 3H), 1.15 - 1.17 (m, 3H).PATENT
[2444] ATTORNEY-DOCKET NO.: 51478-036WO4
[2445] Example 129: 3-((1s,3S)-3-fluorocyclobutyl)-8-((4-isopropylphenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-((1 r,3R)-3-fluorocyclobutyl)-8-((4-isopropylphenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 135 and Compound 136)
[2446] Step 1. Synthesis of 5-amino-2-chloro-N-(3-fluorocyclobutyl)-6-(methylthio) pyrimidine-4-carboxamide
[2447] X «H.?HATH, DiEA, DMF X
[2448] 11: H
[2449] ci' y cr Y
[2450]
[2451] To a solution of 5-amino-2-chloro-6-(methylthio) pyrimidine-4-carboxylic acid (1 g, 4.553 mmol, 1 equiv) and HATU (3.46 g, 9.106 mmol, 2 equiv) in DMF (10 mL) was added DIEA (1.18 g, 9.106 mmol, 2 equiv) at 0°C. The resulting mixture was stirred at room temperature for 5min. To the above mixture was added 3-fluorocyclobutan-1 -amine hydrochloride (0.61 g, 6.829 mmol, 1.5 equiv) at room temperature. The resulting mixture was stirred at room temperature for additional 1h. Desired product could be detected by LCMS. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN and H2O (0.1% TFA), 2% to 50% gradient in 20 min; detector, UV 254 nmto afford 5-amino-2-chloro-N-(3-fluorocyclobutyl)-6-(methylthio) pyrimidine-4-carboxamide (880 mg, 66.48% yield, 90% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C10H12CIFN4OS: 290.04, Found: 291,05[M+H]+.
[2452] Step 2. Synthesis of 6-chloro-3-(3-fluorocyclobutyl)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2453] ^,0 6
[2454] '"'S '"'3
[2455] HDAc X
[2456] 1 ' H * T 1
[2457] J, H N cr 'N' Y 120*0, 5 hrs CL Y 'y'A
[2458]
[2459] O O ' ' ' T To a solution of 5-amino-2-chloro-N-(3-fluorocyclobutyl)-6-(methylthio) pyrimidine-4-carboxamide (500 mg, 1.720 mmol, 1 equiv) in trimethoxymethane (5 mL) was added HOAc (309.8 mg, 5.160 mmol, 3 equiv) at room temperature. The resulting mixture was stirred at 120°C for 5 h. Desired product could be detected by LCMS. The mixture was allowed to cool down to room temperature. The reaction was quenched with ice water at room temperature. The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (1 x 100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 6-chloro-3-(3-fluorocyclobutyl)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (300 mg, 58.00% yield, 92% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C11H10CIFN4OS: 300.02, Found: 301,10[M+H]+.PATENT
[2460] ATTORNEY-DOCKET NO.: 51478-036WO4
[2461] Step 3. Synthesis of (R)-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2462] DiEA. DM-SO
[2463]
[2464] To a solution of 6-chloro-3-(3-fluorocyclobutyl)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (300 mg, 0.998 mmol, 1 equiv) and (R)-2-methylmorpholine (151.3 mg, 1.497 mmol, 1.5 equiv) in DMSO (3 mL) was added DIEA (386.8 mg, 2.994 mmol, 3 equiv) at room temperature. The resulting mixture was stirred at 100°C for 1h. Desired product could be detected by LCMS. The mixture was allowed to cool down to room temperature. The reaction was quenched with ice water at room temperature. The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (1 x 100 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 reverse phase gel; mobile phase, ACN and H2O (0.1% TFA), 2% to 70% gradient in 15 min; detector, UV 254 nm to afford (R)-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (260 mg, 71.32% yield, 95% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C16H20FN5O2S: 365.13, Found: 366.05[M+H]+.
[2465] Step 4. Synthesis of (R)-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino)-8-(methylsulfonyl) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2466] .0
[2467] ij | m-CPSA. DCM
[2468] r N *N" Y - *
[2469] CK J O G" C, 1 Sw
[2470]
[2471] To a solution of (R)-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino)-8-(methylthio) pyrimido[5,4-d] pyrimidin-4(3H)-one (150 mg, 0.410 mmol, 1 equiv) in DCM (2 mL, 31.461 mmol, 76.65 equiv) was added m-CPBA (71 mg, 0.411 mmol, 1.00 equiv) at 0°C. The resulting mixture was stirred at 0°C for 1h. Desired product could be detected by LCMS. The reaction was quenched with ice water (10 mL) at 0°C. The resulting mixture was extracted with CH2CI2 (3 x 80 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN and H2O (0.1% TFA), 2% to 60% gradient in 15 min; detector, UV 254 nm to afford (R)-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino)-8-(methylsulfonyl) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 61.30% yield, 92% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C16H20FN5O4S: 397.12, Found: 398.10[M+H]+.PATENT
[2472] ATTORNEY-DOCKET NO.: 51478-036WO4
[2473] Step 5. Synthesis of (R)-3-(3-fluorocyclobutyl)-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2474] C!
[2475] MeOK, HjO
[2476] ri, I t>r
[2477]
[2478] To a solution of (R)-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino)-8-(methylsulfonyl) pyrimido[5,4-d] pyrimidin-4(3H)-one (100 mg, 0.252 mmol, 1 equiv) in MeOH (1 mL) and H2O (1 mL) was added NaOH (10.1 mg, 0.252 mmol, 1 equiv) at 0 °C. The resulting mixture was stirred at room temperature for 1h. Desired product could be detected by LCMS. The reaction was quenched with ice water at 0°C. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN and H2O (0.1% TFA), 2% to 50% gradient in 15 min; detector, UV 254 nm to afford (R)-3-(3-fluorocyclobutyl)-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (80 mg, 94.81% yield, 92% purity) as a white solid. LC / MS: MS (ESI) calcd. for C15H18FN5O3: 335.14, Found: 336.10[M+H]+.
[2479] Step 6. Synthesis of (R)-8-chloro-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2480] QH Cl
[2481] J JTlV..>1. S N -POGI’ ’*■ JJ N„ r N' Y TA BSTC. 1 hr r Y T k
[2482]
[2483] i I
[2484] A solution of (R)-3-(3-fluorocyclobutyl)-8-hydroxy-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 0.179 mmol, 1 equiv) in POCl3 (2 mL) was stirred at 80°C for 1h. Desired product could be detected by LCMS. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure. The reaction was quenched with ice water at room temperature. The resulting mixture was extracted with EA (3 x 20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford (R)-8-chloro-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, crude product) as a yellow oil. LC / MS: MS (ESI) calcd. for C15H17CIFN5O2: 353.11, Found: 354.10[M+H]+.
[2485] Step 7. Synthesis of (R)-3-(3-fluorocyclobutyl)-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one
[2486]
[2487] To a solution of (R)-8-chloro-3-(3-fluorocyclobutyl)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (60 mg, 0.170 mmol, 1 equiv), 4-isopropylaniline (34.4 mg, 0.255 mmol, 1.5 equiv) in DMSO (2 mL) was added DIEA (65.7 mg, 0.510 mmol, 3 equiv) at room temperature. The resulting mixture was stirred at 100°C for 1h. Desired product could be detected by LCMS. The mixture wasPATENT
[2488] ATTORNEY-DOCKET NO.: 51478-036WO4
[2489] allowed to cool down to room temperature. The reaction was quenched with ice water at room temperature. The resulting mixture was extracted with EA (3 x20 mL). The combined organic layers were washed with brine (1 x 50 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN and H2O (0.1% TFA), 2% to 50% gradient in 15min; detector, UV 254 nm to afford (R)-3-(3-fluorocyclobutyl)-8-((4-isopropylphenyl) amino)-6-(2-methylmorpholino) pyrimido[5,4-d] pyrimidin-4(3H)-one (50 mg, 65.15% yield, 95% purity) as a yellow solid. LC / MS: MS (ESI) calcd. for C24H29FN6O2: 452.23, Found:
[2490] 453.10[M+H]+.
[2491] Step 8. Separation of 3-((1s,3S)-3-fluorocyclobutyl)-8-((4-isopropylphenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one and 3-((1 r,3R)-3-fluorocyclobutyl)-8-((4-isopropylphenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one
[2492]
[2493] The product (70mg) was purified by Chiral-HPLC with the following conditions (Column:
[2494] CHIRALPAK 3; Mobile Phase A: Hex (0.1% DEA): (EtOH: DCM=1: 1)=70: 30; Flow rate: 1.0 mL / min mL / min; Gradient (B%); Injection Volume: 3 mL) to afford the first eluting isomer, 3-((1s,3S)-3-fluorocyclobutyl)-8-((4-isopropylphenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one (13.4 mg, 22.33% yield, 95.0% purity) as a white solid. Stereochemistry arbitrarily assigned.
[2495] LC / MS: MS (ESI) calcd. for C24H29FN6O2: 452.23, Found: 453.15[M+H]+.
[2496] 1H NMR (400 MHz, DMSO-d6) 6 (ppm):9.46 - 9.51 (m, 1H), 8.15 - 8.17 (m, 1H), 7.78 - 7.80 (m, 2H), 7.22 - 7.24 (m, 2H), 5.47 - 5.51 (m, 1H), 5.14 - 5.30 (m, 1H), 4.38 - 4.59 (m, 2H), 3.87 - 3.98 (m, 1 H), 3.42 - 3.61 (m, 2H), 2.81 - 3.08 (m, 4H), 2.61 - 2.80 (m, 3H), 1.18 - 1.23 (m, 9H).
[2497] And also to obtain the second eluting isomer, 3-((1r,3R)-3-fluorocyclobutyl)-8-((4-isopropylphenyl)amino)-6-((R)-2-methylmorpholino)pyrimido[5,4-d]pyrimidin-4(3H)-one trifluoroacetate (13.3 mg, 15.18% yield, 99.9% purity) as a white solid. Stereochemistry arbitrarily assigned.
[2498] LC / MS: MS (ESI) calcd. for C24H29FN6O2: 452.23, Found: 453.15[M+H]+.
[2499] 1H NMR (400 MHz, DMSO-d6) 6 (ppm): 9.46 - 9.51 (m, 1H), 8.16 - 8.21 (m, 1H), 7.73 - 7.85 (m, 2H), 7.16 - 7.28 (m, 2H), 5.30 - 5.51 (m, 1H), 4.36 - 4.52 (m, 3H), 3.87 - 3.98 (m, 1H),3.42 - 3.61 (m, 2H), 2.81 - 3.08 (m, 4H), 2.61 - 2.80 (m, 3H), 1.18 - 1.23 (m, 9H).PATENT
[2500] ATTORNEY-DOCKET NO.: 51478-036WO4
[2501] Example 130: (R)-6-(2-cyclopropylmorpholino)-8-((4-(1 -fluorocyclopropyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one and (S)-6-(2-cyclopropylmorpholino)-8-((4-(1 -fluorocyclopropyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 137 and Compound 138)
[2502] Step 1. Synthesis of 6-(2-cyclopropylmorpholino)-8-((4-(1 -fluorocyclopropyl) phenyl) amino)-3-methylpyrimido[5,4-d] pyrimidin-4(3H)-one
[2503] A F
[2504] Cl L I
[2505] J ' ' NH
[2506] 's>i 1. DffiA. DMSO. SO' G..?<
[2507] Cl ' if - - f-.. N
[2508] O r T Y ''
[2509] 6., J O
[2510]
[2511] 2. D1EA, DMSa, W C, -W
[2512] To a solution of 6,8-dichloro-3-methylpyrimido[5,4-d] pyrimidin-4(3H)-one (53 mg, 0.229 mmol, 1 equiv) and 4-(1 -fluorocyclopropyl) aniline (40 mg, 0.265 mmol, 1.15 equiv) in DMSO (1 mL) was added DIEA (60 mg, 0.464 mmol, 2.02 equiv) at room temperature. The resulting mixture was stirred at 50°C for 0.5 h. Desired product could be detected by LCMS. Next, 2-cyclopropylmorpholine (29 mg, 0.228 mmol, 0.99 equiv) was added to the system at room temperature. The resulting mixture was stirred at 100°C for 0.5 h. Desired product could be detected by LCMS. The residue was purified by reverse-phase flash with the following conditions: Column, C18, 90 g, 20~45pm,100A; mobile phase, CH3CN: H2O (0.05% TFA) = 5%~95% in 25 min; Detector, UV 220 nm & 254 nm to afford the race mate of 6-(2-cyclopropylmorpholino)-8-((4-(1 -fluorocyclopropyl) phenyl) amino)-3-methylpyrimido[5,4-d] pyrimidin-4(3H)-one (51 mg, 50.93% yield, 95.4% purity) as a white solid. LC / MS: MS (ESI) calcd. for C23H25FN6O2: 436.20 m / z, found: 437.15[M+H]+.
[2513] Step 2. Separation of (R)-6-(2-cyclopropylmorpholino)-8-((4-(1-fluorocyclopropyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one and (S)-6-(2-cyclopropylmorpholino)-8-((4-(1 -fluorocyclopropyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one
[2514]
[2515] The product (51 mg) was separated by chiral HPLC with the following conditions: CHIRALPAK IK, 5 pm, 250 mm x 20 mm; Mobile Phase A: MTBE (with 0.5% NH3 (2 M in MeOH)), Mobile Phase B: EtOH; Flow rate: 20 mL / min; Gradient (B%): 25% B; Wave Length: 220 / 254 nm; Sample Solvent: EtOH + DCM; Injection Volume: 0.5 mL; Number Of Runs: 25; Single injection run time(min): 11 to afford (R)-6-(2-cyclopropylmorpholino)-8-((4-(1-fluorocyclopropyl)phenyl)amino)-3-methylpyrimido[5,4-d]pyrimidin-4(3H)-one (11.7 mg, 11.68% yield, 99.8% purity) as a white solid. Stereochemistry arbitrarily assigned.
[2516] RT1(min): 7.49
[2517] LC / MS: MS (ESI) calcd. for C23H25FN6O2: 436.20 m / z, found: 437.15[M+H]+.PATENT
[2518] ATTORNEY-DOCKET NO.: 51478-036WO4
[2519] 1H NMR (300 MHz, DMSO) 6 (ppm): 9.64 - 9.70 (m, 1H), 8.23 (s, 1H), 7.87 - 7.96 (m, 2H), 7.25 -7.33 (m, 2H), 4.56 - 4.64 (m, 1H), 4.35 - 4.45 (m, 1H), 3.91 - 3.98 (m, 1H), 3.43 - 3.55 (m, 3H), 3.40 - 3.43 (m, 1H), 2.94 - 3.09 (m, 1H), 2.72 - 2.91 (m, 2H), 1.40 - 1.55 (m, 2H), 1.12 - 1.19 (m, 2H), 1.04 - 1.09 (m, 1H), 0.48 - 0.58 (m, 2H), 0.25 - 0.42 (m, 2H).
[2520] And also to obtain (S)-6-(2-cyclopropylmorpholino)-8-((4-(1 -fluorocyclopropyl) phenyl) amino)-3-methylpyrimido[5,4-d] pyrimidin-4(3H)-one (13.6 mg, 13.58% yield, 99.5% purity) as a white solid.
[2521] Stereochemistry arbitrarily assigned.
[2522] RT2(min): 8.68
[2523] LC / MS: MS (ESI) calcd. for C23H25FN6O2: 436.20 m / z, found: 437.15[M+H]+.
[2524] 1H NMR (300 MHz, DMSO) 6 (ppm): 9.64 - 9.70 (m, 1H), 8.23 (s, 1H), 7.87 - 7.96 (m, 2H), 7.25 -7.33 (m, 2H), 4.56 - 4.64 (m, 1 H), 4.35 - 4.45 (m, 1 H), 3.91 - 3.98 (m, 1 H), 3.43 - 3.58 (m, 4H), 3.05 - 3.11 (m, 1 H), 2.75 - 2.92 (m, 2H), 1.45 - 1.54 (m, 2H), 1.02 - 1.24 (m, 2H), 0.86 - 0.92 (m, 1 H), 0.46 - 0.56 (m, 2H), 0.21 - 0.43 (m, 2H).
[2525] Example 131: 6-((R)-2-cyclopropylmorpholino)-8-((4-isopropylphenyl)amino)-3-((R)-1 -methylpyrrolidin-3-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one and 6-((S)-2-cyclopropylmorpholino)-8-((4-isopropylphenyl)amino)-3-((R)-1 -methylpyrrolidin-3-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 139 and Compound 140)
[2526] Step 1. Synthesis of (R)-5-amino-2-chloro-6-methoxy-N-(1-methylpyrrolidin-3-yl) pyrimidine-4-carboxamide
[2527] / "'A
[2528] ■ s v
[2529] Q 0
[2530] tlA.. NH.> HATH. D1EA, DMF
[2531] Y >: >.......... „ N A A, OH,. T. -A. fl.....
[2532] cr Y ri, 1 hr ct' N Y
[2533]
[2534] o o A. /
[2535] To a solution of 5-amino-2-chloro-6-methoxypyrimidine-4-carboxylic acid (4.67 g, 22.962 mmol, 1 equiv) and (R)-1-methylpyrrolidin-3-amine (2.3 g, 22.962 mmol, 1 equiv) in DMF (50 mL) were added DIEA (8.90 g, 68.886 mmol, 3 equiv) and HATU (13.10 g, 34.443 mmol, 1.5 equiv). The mixture was stirred at room temperature for 1h. The reaction progress was monitored by LCMS, and it showed the reaction was completed. The reaction mixture was quenched by addition of water (60ml) and extracted with ethyl acetate (50ml x 3). The combined extract was washed with water, brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel, eluted with MeOH / DCM (0-30%) to afford (R)-5-amino-2-chloro-6-methoxy-N-(1-methylpyrrolidin-3-yl) pyrimidine-4-carboxamide (3.5 g, 53.35% yield, 90% purity) as yellow semi-solid. MS: MS (ESI) calcd. for C11H16CIN5O2: 285.10, Found: 286.05 [M+H]+.PATENT
[2536] ATTORNEY-DOCKET NO.: 51478-036WO4
[2537] Step 2. Synthesis of 5-amino-2-(2-cyclopropylmorpholino)-6-methoxy-N-((R)-1-methylpyrrolidin-3-yl) pyrimidine-4-carboxamide
[2538] ' tret
[2539] 6,. J
[2540] '■'? A
[2541] fct ' PdhPEPPSMHeptCj, 3-iiitorapyridine, tBoxane
[2542] £!.:. '. "M7
[2543] cr N 'K v wc,3hrc f 'TT 'N' Y *<%-.• o o. > d
[2544]
[2545] To a solution of (R)-5-amino-2-chloro-6-methoxy-N-(1-methylpyrrolidin-3-yl) pyrimidine-4-carboxamide (0.9 g, 3.150 mmol, 1 equiv) and 2-cyclopropylmorpholine (0.80 g, 6.300 mmol, 2.00 equiv) in dioxane (10 mL) were added Cs₂CO₃ (3.08 g, 9.450 mmol, 3 equiv) and Pd-PEPPSI-IHeptCI 3-chloropyridine (529.27 mg, 0.630 mmol, 0.20 equiv) under nitrogen atmosphere. The resulting mixture was stirred at 105°C for 3 h under nitrogen atmosphere. The residue was purified by C18 reversed phase column chromatography, eluted with MeCN / FLO (with TFA) (10% - 95%) to afford 5-amino-2-(2-cyclopropylmorpholino)-6-methoxy-N-((R)-1-methylpyrrolidin-3-yl) pyrimidine-4-carboxamide (950 mg, 80.12% yield, 90% purity) as a yellow solid. MS: MS (ESI)...
Claims
1. PATENTATTORNEY-DOCKET NO.: 51478-036WO4What is claimed is:CLAIMS1. A compound having the structure of Formula (I):Formula (I),or a pharmaceutically acceptable salt thereof, wherein:R1is optionally substituted C5-C12 aryl, optionally substituted C3-C12 heteroaryl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;R2is H, optionally substituted C1-C4 alkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;R3is H, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;R4is H, optionally substituted C1-C5 alkyl, optionally substituted C1-C5 heteroalkyl, optionally substituted C3-C12 cycloalkyl, or optionally substituted C3-C12 heterocycle;R5is optionally substituted C3-C12 heteroaryl, optionally substituted C3-C12 heterocycle, NR6R7, or OR9;R6is H or C1-C6alkyl;R7is optionally substituted C1-C6 alkyl, C1-C6 heteroalkyl, or optionally substituted C3-C12 heterocycle, wherein the alkyl or heterocycle is optionally substituted with 1 or 2 substituents selected from F, OH, C1-C3 alkoxy, and (CH2)nR8;R8is 5- or 6-membered heteroaryl, 3- to 5-membered cycloalkyl, or 3- to 6-membered heterocycle;R9is C3-C12 heterocycle; andn is 1, 2, or 3.
2. The compound of claim 1, wherein R1is optionally substituted C5-C12 aryl.
3. The compound of claim 2, wherein the optionally substituted C5-C12 aryl isPATENTATTORNEY-DOCKET NO.: 51478-036WO44. The compound of claim 1, wherein R1is optionally substituted C3-C12 heteroaryl.PATENTATTORNEY-DOCKET NO.: 51478-036WO45. The compound of claim 4, wherein the optionally substituted C3-C12 heteroarylis6. The compound of claim 1, wherein R1is optionally substituted C3-C12 heterocycle.
7. The compound of claim 6, wherein the optionally substituted C3-C12 heterocycle is8. The compound of claim 1, wherein R1is optionally substituted C3-C12 cycloalkyl.
9. The compound of claim 8, wherein the optionally substituted C3-C12 cycloalkyl isor10. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- A):Formula (l-A),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO411. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- B):Formula (l-B),or a pharmaceutically acceptable salt thereof.
12. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- C):Formula (l-C),or a pharmaceutically acceptable salt thereof.
13. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- D):Formula (l-D),or a pharmaceutically acceptable salt thereof.
14. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- E):Formula (l-E),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO415. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- F):Formula (l-F),or a pharmaceutically acceptable salt thereof.
16. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- G):Formula (l-G),or a pharmaceutically acceptable salt thereof.
17. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- H):Formula (l-H),or a pharmaceutically acceptable salt thereof.
18. The compound of claim 17, wherein the compound of Formula (l-H) has the structure of Formula (l-l):OFormula (l-l),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO419. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- J):or a pharmaceutically acceptable salt thereof.
20. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- K):Formula (l-K),or a pharmaceutically acceptable salt thereof.
21. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- L):Formula (l-L),or a pharmaceutically acceptable salt thereof.
22. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- M):Formula (l-M),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO423. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- N):Formula (l-N),or a pharmaceutically acceptable salt thereof.
24. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- O):Formula (l-O),or a pharmaceutically acceptable salt thereof.
25. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- P):Formula (l-P),or a pharmaceutically acceptable salt thereof.
26. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- Q):Formula (l-Q),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO427. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- R):^N'R2^N., R3R5NFormula (l-R),or a pharmaceutically acceptable salt thereof.
28. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-S):N\IN'R2R- R5N Y R‘OFormula (l-S),or a pharmaceutically acceptable salt thereof.
29. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- T):N\'R2.. J\, RN:*£ \|R5N y' R‘oFormula (l-T),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO430. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- U):or a pharmaceutically acceptable salt thereof.
31. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- V):Formula (l-V),or a pharmaceutically acceptable salt thereof.
32. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- W):Formula (l-W),or a pharmaceutically acceptable salt thereof.
33. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- X):Formula (l-X),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO434. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- Y):Formula (l-Y),or a pharmaceutically acceptable salt thereof.
35. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- Bl):Formula (l-BI),or a pharmaceutically acceptable salt thereof.
36. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- BJ):Formula (l-BJ),or a pharmaceutically acceptable salt thereof.
37. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- BK):Formula (l-BK),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO438. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- BL):Formula (l-BL),or a pharmaceutically acceptable salt thereof.
39. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AL):Formula (l-AL),or a pharmaceutically acceptable salt thereof.
40. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AM):Formula (l-AM),or a pharmaceutically acceptable salt thereof.
41. The compound of claim 40, wherein the compound of Formula (l-AM) has the structure of Formula (l-AM-1):PATENTATTORNEY-DOCKET NO.: 51478-036WO4or a pharmaceutically acceptable salt thereof.
42. The compound of claim 40, wherein the compound of Formula (l-AM) has the structure of Formula (l-AM-2):CF3N'R2X ^R:R5NN'R-OFormula (l-AM-2),or a pharmaceutically acceptable salt thereof.
43. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AN):N'R2R1¥ 0N'R4Formula (l-AN),or a pharmaceutically acceptable salt thereof.
44. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AU):N'R2X >R1N'R‘Formula (l-AU),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO445. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AV):Formula (l-AV),or a pharmaceutically acceptable salt thereof.
46. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AW):Formula (l-AW),or a pharmaceutically acceptable salt thereof.
47. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AX):Formula (l-AX),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO448. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AY):N'R2R1%OFormula (l-AY),or a pharmaceutically acceptable salt thereof.
49. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AZ):R'R5N y 'R‘oFormula (l-AZ),or a pharmaceutically acceptable salt thereof.
50. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- BA):J-L R'Sr*21. R:NR5^N'xy'N'R‘oFormula (l-BA),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO451. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-Formula (l-BB),or a pharmaceutically acceptable salt thereof.
52. The compound of claim 51, wherein the compound of Formula (l-BB) has the structure of Formula (l-BB-1):Formula (l-BB-1),or a pharmaceutically acceptable salt thereof.
53. The compound of claim 51, wherein the compound of Formula (l-BB) has the structure of Formula (l-BB-2):Formula (l-BB-2),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO454. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- BC):OFormula (l-BC),or a pharmaceutically acceptable salt thereof.
55. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-0Formula (l-BD),or a pharmaceutically acceptable salt thereof.
56. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-Formula (l-BE),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO457. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BF):NN'R2k ^NK ^R:R5NN'R‘OFormula (l-BF),or a pharmaceutically acceptable salt thereof.
58. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BG):^N'R2R5NOFormula (l-BG),or a pharmaceutically acceptable salt thereof.
59. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BH):■L'R2^N.. R:R5NOFormula (l-BH),or a pharmaceutically acceptable salt thereof.
60. The compound of any one of claims 1 to 59, wherein R2is H.
61. The compound of any one of claims 1 to 59, wherein R2is optionally substituted C1-C4 alkyl.
62. The compound of claim 61, wherein the optionally substituted C1-C4 alkyl isCH3.
63. The compound of any one of claims 1 to 62, wherein R3is H.PATENTATTORNEY-DOCKET NO.: 51478-036WO464. The compound of any one of claims 1 to 62, wherein R3is optionally substituted C1-C4 alkyl.
65. The compound of claim 64, wherein the optionally substituted C1-C4 alkyl isun3,1\, orF66. The compound of any one of claims 1 to 65, wherein R4is H.
67. The compound of any one of claims 1 to 65, wherein R4is optionally substituted C1-C5 alkyl.
68. The compound of claim 67, wherein the optionally substituted C1-C5 alkyl isCH369. The compound of any one of claims 1 to 65, wherein R4is optionally substituted C1-C5 heteroalkyl.N 70. The compound of claim 69, wherein the optionally substituted C1-C5 heteroalkyl is I or71. The compound of any one of claims 1 to 65, wherein R4is optionally substituted C3-C12 heterocycle.
72. The compound of claim 71, wherein the optionally substituted C3-C12 heterocycle is73. The compound of any one of claims 1 to 65, wherein R4is optionally substituted C3-C12 cycloalkyl.PATENTATTORNEY-DOCKET NO.: 51478-036WO474. The compound of claim 73, wherein the optionally substituted C3-C12 cycloalkyl is75. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- Z):Formula (l-Z),or a pharmaceutically acceptable salt thereof.
76. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AA):Formula (l-AA),or a pharmaceutically acceptable salt thereof.
77. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BM):Formula (l-BM),or a pharmaceutically acceptable salt thereof.
78. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BN):RVR2Formula (l-BN),PATENTATTORNEY-DOCKET NO.: 51478-036WO4or a pharmaceutically acceptable salt thereof.
79. The compound of claim 78, wherein the compound of Formula (l-BN) has the structure of Formula (l-BN-1):Formula (l-BN-1),or a pharmaceutically acceptable salt thereof.
80. The compound of claim 78, wherein the compound of Formula (l-BN) has the structure of Formula (l-BN-2):R!N-R2oFormula (l-BN-2),or a pharmaceutically acceptable salt thereof.
81. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AB):R?N'R1Formula (l-AB),or a pharmaceutically acceptable salt thereof.
82. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AO):Formula (l-AO),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO483. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AP):Formula (l-AP),or a pharmaceutically acceptable salt thereof.
84. The compound of claim 83, wherein the compound of Formula (l-AP) has the structure of Formula (l-AP-1):R5N-R1Formula (l-AP-1),or a pharmaceutically acceptable salt thereof.
85. The compound of claim 83, wherein the compound of Formula (l-AP) has the structure of Formula (l-AP-2):Formula (l-AP-2),or a pharmaceutically acceptable salt thereof.
86. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BO):Formula (l-BO),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO487. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BP):R5N'R1Formula (l-BP),or a pharmaceutically acceptable salt thereof.
88. The compound of claim 87, wherein the compound of Formula (l-BP) has the structure of Formula (l-BP-1):R?N'R1Formula (l-BP-1),or a pharmaceutically acceptable salt thereof.
89. The compound of claim 87, wherein the compound of Formula (l-BP) has the structure of Formula (l-BP-2):Formula (l-BP-2),or a pharmaceutically acceptable salt thereof.
90. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BQ):R-N-R’Formula (l-BQ),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO491. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BR):R;N-R1Formula (l-BR),or a pharmaceutically acceptable salt thereof.
92. The compound of any one of claims 1 to 91, wherein R5is optionally substituted C3-C12 heteroaryl.
94. The compound of any one of claims 1 to 91, wherein R5is optionally substituted C3-C12 heterocycle.
95. The compound of claim 94, wherein the optionally substituted C3-C12 heterocycle isPATENTATTORNEY-DOCKET NO.: 51478-036WO496. The compound of any one of claims 1 to 91, wherein R5is NR6R7.
97. The compound of claim 96, wherein the NR6R7is98. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AC):Formula (l-AC),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO499. The compound of claim 98, wherein the compound of Formula (l-AC) has the structure of Formula (l-AD):Formula (l-AD),or a pharmaceutically acceptable salt thereof.
100. The compound of claim 98, wherein the compound of Formula (l-AC) has the structure of Formula (l-AE):Formula (l-AE),or a pharmaceutically acceptable salt thereof.
101. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BS):Formula (l-BS),or a pharmaceutically acceptable salt thereof.
102. The compound of claim 101, wherein the compound of Formula (l-BS) has the structure of Formula (l-BS-1):R\,, R2Formula (l-BS-1),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO4103. The compound of claim 101, wherein the compound of Formula (l-BS) has the structure of Formula (l-BS-2):Formula (l-BS-2),or a pharmaceutically acceptable salt thereof.
104. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BT):Formula (l-BT),or a pharmaceutically acceptable salt thereof.
105. The compound of claim 104, wherein the compound of Formula (l-BT) has the structure of Formula (l-BT-1):Formula (l-BT-1),or a pharmaceutically acceptable salt thereof.
106. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BU):Formula (l-BU),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO4107. The compound of claim 106, wherein the compound of Formula (l-BU) has the structure of Formula (l-BU-1):Formula (l-BU-1),or a pharmaceutically acceptable salt thereof.
108. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BV):R1.,, R2Formula (l-BV),or a pharmaceutically acceptable salt thereof.
109. The compound of claim 108, wherein the compound of Formula (l-BV) has the structure of Formula (l-BV-1):R,-N-R2Formula (l-BV-1),or a pharmaceutically acceptable salt thereof.
110. The compound of claim 108, wherein the compound of Formula (l-BV) has the structure of Formula (l-BV-2):R1,, R2Formula (l-BV-2),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO4111. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BW):N N N R'OFormula (l-BW),or a pharmaceutically acceptable salt thereof.
112. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- BX):i T ^ VNxp N N R'O^ J OFormula (l-BX),or a pharmaceutically acceptable salt thereof.
113. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BY):N^bkp N N j^ R‘N. J OFormula (l-BY),or a pharmaceutically acceptable salt thereof.
114. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-BZ):^NxF < N N R'-L ^bk J OFormula (l-BZ),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO4115. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AF):Formula (l-AF),or a pharmaceutically acceptable salt thereof.
116. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AQ):Formula (l-AQ),or a pharmaceutically acceptable salt thereof.
117. The compound of claim 116, wherein the compound of Formula (l-AQ) has the structure of Formula (l-AQ-1):Formula (l-AQ-1),or a pharmaceutically acceptable salt thereof.
118. The compound of claim 116, wherein the compound of Formula (l-AQ) has the structure of Formula (l-AQ-2):Formula (l-AQ-2),PATENTATTORNEY-DOCKET NO.: 51478-036WO4or a pharmaceutically acceptable salt thereof.
119. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I- AR):Formula (l-AR),or a pharmaceutically acceptable salt thereof.
120. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AG):Formula (l-AG),or a pharmaceutically acceptable salt thereof.
121. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AH):Formula (l-AH),or a pharmaceutically acceptable salt thereof.
122. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AI):Formula (l-AI),or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO4123. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AJ):Formula (l-AJ),or a pharmaceutically acceptable salt thereof.
124. The compound of claim 1, wherein the compound of Formula (I) has the structure of Formula (I-AK):Formula (l-AK),or a pharmaceutically acceptable salt thereof.
125. The compound of claim 1, wherein the compound of Formula (I) has the structure:PATENTATTORNEY-DOCKET NO.: 51478-036WO4Formula (ll-P),Formula (11-R),PATENTATTORNEY-DOCKET NO.: 51478-036WO4Formula (ll-X),Formula (ll-AM-2),Formula (ll-AW),PATENTATTORNEY-DOCKET NO.: 51478-036WO4Formula (ll-BB-1),Formula (ll-BG),Formula (ll-BH), Formula (ll-BI),PATENTATTORNEY-DOCKET NO.: 51478-036WO4Formula (ll-BK),Formula (ll-BL), or a pharmaceutically acceptable salt thereof.
126. The compound of claim 1, wherein the compound of Formula (I) has the structure:Formula (ll-AB), or a pharmaceutically acceptable salt thereof.
127. The compound of claim 1, wherein the compound of Formula (I) has the structure:Formula (ll-AF),PATENTATTORNEY-DOCKET NO.: 51478-036WO4Formula (II-AQ), Formula (II-AQ-1),Formula (ll-BV),Formula (ll-BV-2),PATENTATTORNEY-DOCKET NO.: 51478-036WO4pharmaceutically acceptable salt thereof.
128. The compound of claim 1, wherein the compound of Formula (I) has the structure:Formula (ll-AI), Formula (ll-AK), or a pharmaceutically acceptable salt thereof.
129. The compound of claim 1, wherein the compound of Formula (I) has the structure:Formula (ll-AT), or a pharmaceutically acceptable salt thereof.PATENTATTORNEY-DOCKET NO.: 51478-036WO4130. The compound of claim 1, wherein the compound of Formula (I) has the structure:Formula (ll-BR), or a pharmaceutically acceptable salt thereof.
131. The compound of claim 1, wherein the compound has the structure of any of compounds 1 -210 in Table 1, or a pharmaceutically acceptable salt thereof.
132. A pharmaceutical composition comprising a compound of any one of claims 1 to 131 and a pharmaceutically acceptable excipient.PATENTATTORNEY-DOCKET NO.: 51478-036WO4133. A compound of any one of claims 1 to 131, or a pharmaceutical composition of claim 132 for use as a medicament.
134. A method of treating or preventing pain in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 131, or a pharmaceutical composition of claim 132.
135. The method of claim 134, wherein the pain is neuropathic pain, inflammation, chronic pain, inflammatory pain, arthritic pain, diabetic pain, post-operative pain, lower back pain, central pain, fibromyalgia or neuralgia.
136. A method of treating epilepsy in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 131, or a pharmaceutical composition of claim 132.
137. The method of claim 136, wherein the epilepsy is refractory epilepsy, developmental and epileptic encephalopathies, Lennox-Gastaut syndrome, neurotrauma associated epilepsy, status epilepticus, tumor associated epilepsy, or hypoxic-ischemic encephalopathy.
138. A method of treating a neurodevelopmental disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any one of claims 1 to 131, ora pharmaceutical composition of claim 132.
139. The method of claim 138, wherein the neurodevelopmental disorder is autism spectrum disorder, Rett Syndrome, Tuberous Sclerosis Complex, Fragile X syndrome, Angelman syndrome, Down syndrome, Dravet syndrome, CKDL5 Deficiency syndrome, SYNGAP1, 22q11.2 microdeletion syndrome, or cerebral palsy.
140. A method of treating neurotraumatic injury in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 131, or a pharmaceutical composition of claim 132.
141. The method of claim 140, wherein the neurotraumatic injury is traumatic brain injury, stroke, nerve injury, or spinal cord injury.
142. A method of treating neurodegenerative disorder in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 131, ora pharmaceutical composition of claim 132.PATENTATTORNEY-DOCKET NO.: 51478-036WO4143. The method of claim 142, wherein the neurodegenerative disorder is multiple sclerosis, amyotrophic lateral sclerosis, Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Lewy body dementia, or frontotemporal dementia.
144. A method of treating an affective disorder in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 131, or a pharmaceutical composition of claim 132.
145. The method of claim 144, wherein the affective disorder is schizophrenia, bipolar disorder, general anxiety disorder, social anxiety disorder, attention-deficit / hyperactivity disorder, or major depressive disorder.
146. A method of treating dementia in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 131, or a pharmaceutical composition of claim 132.
147. The method of claim 146, wherein the dementia is frontotemporal dementia or Lewy body dementia.
148. A method of treating dementia-induced psychosis in a subject in need thereof, the method including administering to the subject an effective amount of a compound of any one of claims 1 to 131, ora pharmaceutical composition of claim 132.
149. A method for potentiating KCC2 activity, clustering, dimerization or membrane expression in a cell or subject, the method comprising contacting the cell with, or administering to the subject, an effective amount of a compound of any one of claims 1 to 131, or a pharmaceutical composition of claim 132.
150. A method for increasing Cl efflux or potentiating KCC2 activity in a cell or subject, the method comprising contacting the cell with, or administering to the subject, an effective amount of a compound of any one of claims 1 to 131, ora pharmaceutical composition of claim 132.