, Yield: 14%; 14 mg; Yellow solid, 8H)-one (4a), vol.3068, pp.699-700, 1346.
H Ar ), 3.73 (3H, s, CH 3 ), CDCl 3 , 25 ? C, 300 MHz): ? H 9.39 (1H, br s, H Ar ), 8.73 (1H, d, J = 4.8 Hz, H Ar ), 8.58-8.36 (2H, m, H Ar + H 4 ), 8.19 (1H, s, H 7 ), vol.7, pp.129-154 ,
, 95:5; v/v); mp 202-205 ? C; IR (neat) ? max 3540, HRMS (ESI + ): Calcd for C 15 H 10 N 4 OS, pp.34-37
, 8H)-one (4c or FC162) [34]: yield: 69%; 91 mg
, CDCl 3 , 25 ? C, 300 MHz): ? H 9.41 (d, J = 2.3 Hz, 1H, H Ar ), 8.75 (d, J = 4.9 Hz, 1H, H Ar ), 8.54-8.38 (m, 2H, H Ar + H 4 ), 8.26 (s, 1H, H 7 ), 7.88 (d, J = 8.7 Hz, 1H, H 5 ), vol.3012, pp.151-160, 1024.
, Cyclobutyl-2-(pyridin-3-yl)thiazolo[5,4-f]quinazolin-9(8H)-one (4d): Yield: 57%; 64 mg
, 39 (1H, br s, H Ar ), 8.72 (1H, d, J = 4.8, H Ar ), 8.45-8.29 (2H, m, H Ar + H 4 ), 8.30 (1H, s, H 7, CDCl 3 + D 2 O, 25 ? C, 300 MHz): ? H 9, vol.7, pp.848-849, 1347.
95:5; v/v); mp 212-215 ? C; IR (neat) ? max 3064, 3964, 2872; 1902, 1645, 1584, 1451, 828 cm ?1 ; 1 H-NMR (CDCl 3 , 25 ? C, 300 MHz): ? H 9.40 (1H, br s, H Ar ) ,
5 (CH), 148.4 (CH), 146.3, 144.1 (CH), ? C 168.0 (C), 159.8 (C), vol.152, p.151 ,
, Calcd for C 19 H 16 N 4 OS
CDCl 3 , 25 ? C, 300 MHz): ? H 9.38 (1H, br s, H Ar ), 8.72 (1H, br s, H Ar ), 2H, m, H Ar + H 4 ), 8.25 (1H, s, H 7 ), 7.84 (1H, d, J = 8.7 Hz, H 4, vol.3146, pp.2181-2194, 1076. ,
, × CH 2 ), 25.2 (CH 2 ). HRMS (ESI + ): Calcd for, vol.54
, In Vitro Kinase Preparation and Assays
, Homogenization buffer: 25 mM MOPS; 15 mM EGTA; 15 mM MgCl 2 ; 60 mM ?-glycerophosphate
, Buffer A: 10 mM MgCl 2 ; 1 mM EGTA; 1 mM DTT; 25 mM Tris/HCl, and 50 µg/mL heparin. Buffer B: 60 mM ?-glycerophosphate; 30 mM p-nitrophenylphosphate; 25 mM MOPS pH 7.0; sodium vanadate. All chemicals were purchased from Sigma-Aldrich, vol.2, p.15
, Kinase Preparations and Assays Kinase activities were assayed in triplicates, in buffer A or B, at 30 ? C, at a final adenosine triphosphate (ATP) concentration of 15 µmol/L. Blank values were subtracted, and activities were expressed in percent (%) of the maximal activity
, Ci/mmol; 10 mCi/mL) in a final volume of 30 µL. After 30 min incubation at 30 ? C, 25 µL aliquots of supernatant were spotted onto sheets of Whatman P81 phosphocellulose paper, and 20 s later, the filters were washed eight times (for at least 5 min each time) in a solution of 10 mL phosphoric acid/L of water. The wet filters were counted in the presence of 1 mL ACS (Amersham) scintillation fluid. GSK-3?/? (porcine brain, native) was assayed, in buffer A, with 0.5 mg BSA/mL + 1 mM DTT, using GS-1 (YRRAAVPPSPSLSRHSSPHQSpEDEEE) (pS stands for phosphorylated serine), a GSK-3 specific substrate, Its kinase activity was assayed in buffer A, with 1 mg of histone H1/mL, in the presence of 15 µM
, native) was assayed as described for CDK1, but in buffer B, and using 25 µM CKS peptide (RRKHAAIGpSAYSITA), a CK1-specific substrate, vol.39
, DYRK1A (Human, recombinant, expressed in E. coli as GST fusion proteins) was purified by affinity chromatography on glutathione-agarose and assayed as described for CDK1/cyclin B, with with 0.5 mg BSA/mL + 1 mM DTT and using Woodtide (KKISGRLSPIMTEQ) (1.5 µg/assay) as a substrate
, Conclusions This work demonstrates the efficacy of synthetic methodologies, such as C-H arylation of arenes and hetero-arenes for SAR studies. The application of this powerful tool at the last stage of the synthesis of kinase inhibitors allowed the synthesis of arrays of molecules inspired by fragment-growing studies generated by molecular modeling calculations. Among the potential active compounds generated through this strategy, vol.162
, Supplementary Materials: The following are available online. 1 H-NMR and 13 C-NMR spectra of new compounds 8a-f and 4a-f
conceived the project and designed the experiments. F.C. and M.H. performed the experimental work ,
designed and supervised the biological experiments. T.B. wrote the manuscript with the cooperation of C.F. All authors discussed the results and commented on the manuscript ,
thank the LABEX SynOrg (ANR-11-LABX-0029) for financial support. This research was supported by grants from the, Fonds Unique Interministériel" (FUI) TRIAD project and Conseil Régional de Bretagne (L.M.) and the "Fondation Jérôme Lejeune ,
Synthesis of novel pentacyclic pyrrolothiazolobenzoquinolinones, analogs of natural marine alkaloids, Tetrahedron Lett, vol.42, pp.2673-2676, 2001. ,
Synthesis and cytotoxic evaluation of novel thiazolocarbazoles, J. Enz. Inhib. Med. Chem, vol.17, pp.397-401, 2002. ,
Novel 6-substituted benzothiazol-2-yl indolo[1,2-c]quinazolines and benzimidazo[1,2-c]quinazolines, Tetrahedron, vol.59, pp.773-779, 2003. ,
Synthesis and cytotoxic activity of thiazolofluorenone derivatives, J. Enzym. Inhib. Med. Chem, vol.19, pp.567-575, 2004. ,
Microwave-assisted synthesis of novel thiazolocarbazoles and evaluation as potential anticancer agents. Part III, J. Enz. Inhib. Med. Chem, vol.19, pp.467-473, 2004. ,
URL : https://hal.archives-ouvertes.fr/hal-01967795
Novel 9-oxo-thiazolo[5,4-f ]quinazoline-2-carbonitrile derivatives as dual cyclin-dependent kinase 1 (CDK1)/glycogen synthase kinase-3 (GSK-3) inhibitors: Synthesis, biological evaluation and molecular modeling studies, Eur. J. Med. Chem, vol.43, pp.1469-1477, 2008. ,
Thiazolo[5,4-f ]quinazolin-9-ones, inhibitors of glycogen synthase kinase-3, Bioorg. Med. Chem. Lett, vol.16, pp.3419-3423, 2006. ,
] pyrimidin-4-amines as multitarget Ser/Thr kinases inhibitors, Eur. J. Med. Chem, vol.92, pp.124-134, 2015. ,
]pyrimidin-4-amine analogues as dual inhibitors of CLK1 and DYRK1A kinases, Eur. J. Med. Chem, vol.59, pp.283-295, 2013. ,
URL : https://hal.archives-ouvertes.fr/hal-00997416
Synthesis and biological evaluation of N-arylbenzo[b]thieno[3,2-d]pyrimidin-4-amines and their pyrido and pyrazino analogues as Ser/Thr kinase inhibitors, Eur. J. Med. Chem, vol.58, pp.171-183, 2012. ,
URL : https://hal.archives-ouvertes.fr/hal-00997084
Microwave-accelerated Dimroth rearrangement for the synthesis of 4-anilino-6-nitroquinazolines. Application to an efficient synthesis of a microtubule destabilizing agent, Tetrahedron, vol.66, pp.4495-4502, 2010. ,
URL : https://hal.archives-ouvertes.fr/hal-01020796
Design and synthesis of thiazolo[5,4-f ]quinazolines as DYRK1A inhibitors, Part I, Molecules, vol.19, pp.15546-15571, 2014. ,
URL : https://hal.archives-ouvertes.fr/hal-01141172
Design and synthesis of thiazolo[5,4-f ]quinazolines as DYRK1A inhibitors, Part II, Molecules, vol.19, pp.15411-15439, 2014. ,
URL : https://hal.archives-ouvertes.fr/hal-01135223
DYRK1 inhibitors and uses thereof WO 2013026806, Chem. Abstr, vol.158, p.390018, 2013. ,
Synthesis of Thiazolo[5,4-f ]quinazolin-9(8H)ones as Multi-Target Directed Ligands of Ser/Thr Kinases, Molecules, vol.21, 2016. ,
Synthesis of Bioactive 2-(Arylamino)thiazolo[5,4-f ]-quinazolin-9-ones via the Hügershoff Reaction or Cu-Catalyzed Intramolecular C-S Bond Formation, Molecules, vol.21, 2016. ,
Tau protein kinases: Involvement in Alzheimer's disease, Ageing Res. Rev, vol.12, pp.289-309, 2013. ,
Regulation of Alzheimer's disease amyloid-? formation by casein kinase I, Proc. Nat. Acad. Sci, vol.104, pp.4159-4164, 2007. ,
Drug discovery process for kinase Inhibitors, Chembiochem, vol.6, pp.455-459, 2005. ,
Small-molecule kinase inhibitors: An analysis of FDA-approved drugs, Drug Discovery Today, vol.21, pp.5-10, 2016. ,
Annotated and updated database of protein kinase inhibitors in clinical trials, Molecules, vol.23, p.908, 2018. ,
Dual-dspecificity tyrosine phosphorylationregulated kinase 1A (DYRK1A) inhibitors as potential therapeutics, J. Med. Chem, p.61, 2018. ,
Dual-specificity tyrosine phosphorylation-regulated kinase 1a (dyrk1a) inhibitors: A survey of recent patent literature, Expert Opin. Ther. Pat, vol.27, pp.1183-1199, 2017. ,
URL : https://hal.archives-ouvertes.fr/hal-02046302
DYRK1A in neurodegeneration and cancer: Molecular basis and clinical implications, Pharmacol. Ther, vol.151, pp.87-98, 2015. ,
Beyond secretases: Kinase inhibitors for the treatment of Alzheimer's disease, Annu. Rep. Med. Chem, vol.48, pp.57-71, 2013. ,
Recent advances in the design, synthesis, and biological evaluation of selective DYRK1A inhibitors: A new avenue for a disease modifying treatment of Alzheimer's?, ACS Chem. Neurosci, vol.3, pp.857-872, 2012. ,
The protein interaction landscape of the human CMGC kinase group, Cell Rep, vol.3, pp.1306-1320, 2013. ,
, Molecules, vol.23, pp.2181-2197, 2018.
A Novel DYRK1A (Dual specificity tyrosine phosphorylationregulated kinase 1A) inhibitor for the treatment of Alzheimer's disease: Effect on Tau and amyloid pathologies in vitro, J. Neurochem, vol.133, pp.440-451, 2015. ,
An unusual binding mode of the methyl 9-anilinothiazolo[5,4-f ]quinazoline-2carbimidates (EHT 1610 and EHT 5372) confers high selectivity for DYRK kinases, J. Med. Chem, vol.59, pp.10315-10321, 2016. ,
Frags2Drugs: An in silico tool to discover new molecules based on 3D fragment network, J. Chem. Inf. Model, 2018. ,
Late-stage C-H Pd-catalyzed and copper assisted regioselective sequential C2 and C7 arylation of thiazolo[5,4-f ]quinazolin-9(8H)-one with aryl halides, Org. Lett, vol.18, pp.3282-3285, 2016. ,
Arylation of thiazolo[5,4-f ]quinazolin-9(8H)-one backbone: Synthesis of an array of potential kinase inhibitors, vol.49, pp.4615-4622, 2017. ,
URL : https://hal.archives-ouvertes.fr/hal-02046272
Recent developments in microwave-assisted metal-catalyzed C-H functionalization of heteroarenes for medicinal chemistry and material applications, Synthesis, vol.48, pp.3879-3889, 2016. ,
URL : https://hal.archives-ouvertes.fr/hal-02046186
Synthese und reaktionen des 4,5-dichlor-1,2,3-dithiazoliumchlorids, Eur. J. Inorg. Chem, vol.118, pp.1632-1643, 1985. ,
Utilization of operational schemes for analog synthesis in drug design, J. Med. Chem, vol.15, pp.1006-1011, 1972. ,
]dec-5-ene (TBD) as a Lewis Base, Synlett, vol.5, issue.1, pp.894-895, 2014. ,
cocrystal structures and neuroprotective properties of leucettines, a family of protein kinase inhibitors derived from the marine sponge alkaloid leucettamine B, J. Med. Chem, vol.55, pp.9312-9330, 2012. ,
URL : https://hal.archives-ouvertes.fr/hal-00870472
Purification of GSK-3 by affinity chromatography on immobilized axin, Protein Expr. Purif, vol.20, pp.394-404, 2000. ,
Purification of CK1 by affinity chromatography on immobilised axin, Protein Expr. Purif, vol.54, pp.101-109, 2007. ,
URL : https://hal.archives-ouvertes.fr/hal-00169405
A review on medicinal importance, pharmacological activity and bioanalytical aspects of beta-carboline alkaloid, Harmine". Asian Pac. J. Trop. Biomed, vol.2, pp.660-664, 2012. ,
, Sample Availability: Samples of compound FC162 (4c) are available from the authors for academic studies with Material Transfer Agreement (MTA)