title: Discovery, synthesis, and characterization of an orally bioavailable, brain penetrant inhibitor of mixed lineage kinase 3.
abstract: Inhibition of mixed lineage kinase 3 (MLK3) is a potential strategy for treatment of Parkinson's disease and HIV-1 associated neurocognitive disorders (HAND), requiring an inhibitor that can achieve significant brain concentration levels. We report here URMC-099 (1) an orally bioavailable (F = 41%), potent (IC50 = 14 nM) MLK3 inhibitor with excellent brain exposure in mouse PK models and minimal interference with key human CYP450 enzymes or hERG channels. The compound inhibits LPS-induced TNFα release in microglial cells, HIV-1 Tat-induced release of cytokines in human monocytes and up-regulation of phospho-JNK in Tat-injected brains of mice. Compound 1 likely functions in HAND preclinical models by inhibiting multiple kinase pathways, including MLK3 and LRRK2 (IC50 = 11 nM). We compare the kinase specificity and BBB penetration of 1 with CEP-1347 (2). Compound 1 is well tolerated, with excellent in vivo activity in HAND models, and is under investigation for further development.
authors.affiliation: Califia Bio Inc. , 11575 Sorrento Valley Road, San Diego 92121, California, United States.
authors.forename: Val S
authors.initials: VS
authors.lastname: Goodfellow
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Loweth
authors.forename: Satheesh B
authors.initials: SB
authors.lastname: Ravula
authors.forename: Torsten
authors.initials: T
authors.lastname: Wiemann
authors.forename: Thong
authors.initials: T
authors.lastname: Nguyen
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Todd
authors.forename: David
authors.initials: D
authors.lastname: Sheppard
authors.forename: Scott
authors.initials: S
authors.lastname: Pollack
authors.forename: Oksana
authors.initials: O
authors.lastname: Polesskaya
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Marker
authors.forename: Stephen
authors.initials: S
authors.lastname: Dewhurst
authors.forename: Harris A
authors.initials: HA
authors.lastname: Gelbard
mesh_terms: D000284:Administration, Oral; D000818:Animals; D019540:Area Under Curve; D001682:Biological Availability; D001812:Blood-Brain Barrier / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D002227:Carbazoles / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000494:pharmacology; D002478:Cells, Cultured; D003072:Cognition Disorders / Q000150:complications / Q000517:prevention & control; D055808:Drug Discovery / Q000379:methods*; D015658:HIV Infections / Q000150:complications; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D020930:MAP Kinase Kinase Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008956:Models, Chemical; D015394:Molecular Structure; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000494:pharmacology*; D011725:Pyridines / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000494:pharmacology*; D011758:Pyrroles / Q000138:chemical synthesis / Q000493:pharmacokinetics / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000494:pharmacology; D000097767:Mitogen-Activated Protein Kinase Kinase Kinase 11
chemical_list: D002227:Carbazoles; D048031:JNK Mitogen-Activated Protein Kinases; D020930:MAP Kinase Kinase Kinases; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011758:Pyrroles; D014409:Tumor Necrosis Factor-alpha; D054322:tat Gene Products, Human Immunodeficiency Virus; D000097767:Mitogen-Activated Protein Kinase Kinase Kinase 11; C583037:URMC-099; C106592:3,9-bis((ethylthio)methyl)-K-252a
references.citation: J Biol Chem. 1998 Jul 10;273(28):17852-8
references.pmid: 9651389
references.citation: J Biol Chem. 2007 Oct 19;282(42):30393-405
references.pmid: 17711861
references.citation: Nat Rev Mol Cell Biol. 2002 Sep;3(9):663-72
references.pmid: 12209126
references.citation: J Med Chem. 2011 Jul 28;54(14):5131-43
references.pmid: 21699136
references.citation: ACS Chem Neurosci. 2011 Jan 19;2(1):3-4
references.pmid: 22778853
references.citation: J Med Chem. 1997 Jun 6;40(12):1863-9
references.pmid: 9191963
references.citation: Int J Exp Pathol. 2007 Apr;88(2):111-26
references.pmid: 17408454
references.citation: J Immunol. 2006 Jul 1;177(1):702-11
references.pmid: 16785569
references.citation: Blood. 2010 Feb 11;115(6):1131-6
references.pmid: 20008298
references.citation: Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8116-20
references.pmid: 9223324
references.citation: Nat Biotechnol. 2005 Mar;23(3):329-36
references.pmid: 15711537
references.citation: PLoS Comput Biol. 2009 Jul;5(7):e1000446
references.pmid: 19629159
references.citation: Adv Neuroimmunol. 1994;4(3):199-206
references.pmid: 7874388
references.citation: Int J Hematol. 2013 Jun;97(6):683-94
references.pmid: 23613268
references.citation: J Neurovirol. 2011 Feb;17(1):3-16
references.pmid: 21174240
references.citation: J Biol Chem. 2001 Jul 6;276(27):25302-8
references.pmid: 11325962
references.citation: J Immunol. 2010 Jan 15;184(2):746-56
references.pmid: 19966207
references.citation: J Neurovirol. 2013 Jun;19(3):254-60
references.pmid: 23737347
references.citation: Ann Neurol. 1993 Jun;33(6):576-82
references.pmid: 8498837
references.citation: J Virol. 1998 Jul;72(7):6131-7
references.pmid: 9621077
references.citation: J Biol Chem. 1998 Dec 4;273(49):32408-15
references.pmid: 9829970
references.citation: J Immunol. 1999 Sep 1;163(5):2953-9
references.pmid: 10453044
references.citation: Neuroscience. 2005;135(3):781-90
references.pmid: 16111829
references.citation: Ann Neurol. 1992 Apr;31(4):349-60
references.pmid: 1586135
references.citation: Exp Mol Pathol. 2010 Oct;89(2):109-16
references.pmid: 20566414
references.citation: J Neurosci. 2013 Jun 12;33(24):9998-10010
references.pmid: 23761895
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13795-800
references.pmid: 12374865
references.citation: Neurology. 2007 Oct 9;69(15):1480-90
references.pmid: 17881719
references.citation: J Neurosci. 2005 Jul 6;25(27):6329-42
references.pmid: 16000623
references.citation: J Neuroimmunol. 2004 Dec;157(1-2):66-70
references.pmid: 15579282
references.citation: J Immunol. 2004 Aug 15;173(4):2762-70
references.pmid: 15294995
references.citation: Brain Res. 2004 Apr 2;1003(1-2):86-97
references.pmid: 15019567
references.citation: Expert Opin Drug Discov. 2008 Jun;3(6):607-621
references.pmid: 19662101
references.citation: J Pharmacol Exp Ther. 2006 Dec;319(3):998-1008
references.pmid: 16946104
references.citation: PLoS One. 2011 Apr 08;6(4):e18633
references.pmid: 21494611
references.citation: Curr Neurol Neurosci Rep. 2012 Jun;12(3):251-60
references.pmid: 22441981
references.citation: Curr Top Med Chem. 2008;8(18):1555-72
references.pmid: 19075767
references.citation: J Med Chem. 2008 Sep 25;51(18):5680-9
references.pmid: 18714982
references.citation: Nat Biotechnol. 2011 Oct 30;29(11):1046-51
references.pmid: 22037378
references.citation: J Immunol. 1998 Jul 15;161(2):776-81
references.pmid: 9670954
references.citation: Nat Biotechnol. 2008 Jan;26(1):127-32
references.pmid: 18183025
references.citation: J Med Chem. 2010 Feb 25;53(4):1413-37
references.pmid: 20166671
references.citation: J Exp Med. 1992 Dec 1;176(6):1703-18
references.pmid: 1460427
references.citation: Mol Cell. 2002 Dec;10(6):1527-33
references.pmid: 12504027
references.citation: J Immunol. 2007 Mar 1;178(5):3226-36
references.pmid: 17312171
references.citation: J Neurochem. 2002 Sep;82(6):1424-34
references.pmid: 12354290
references.citation: Annu Rev Pharmacol Toxicol. 2004;44:451-74
references.pmid: 14744254
references.citation: Mov Disord. 2005 Jun;20(6):653-64
references.pmid: 15719422
references.citation: J Neuroinflammation. 2012 Nov 29;9:261
references.pmid: 23190742
references.citation: Mol Cell. 2007 Aug 3;27(3):498-508
references.pmid: 17679097
references.citation: J Biol Chem. 2001 Jan 19;276(3):1961-7
references.pmid: 11053428
references.citation: J Pharmacol Exp Ther. 1999 Feb;288(2):421-7
references.pmid: 9918541
references.citation: Int Immunol. 2007 Aug;19(8):923-33
references.pmid: 17698565
references.citation: J Neurosci. 2011 Mar 2;31(9):3148-57
references.pmid: 21368026
references.citation: Nat Rev Drug Discov. 2009 Nov;8(11):892-909
references.pmid: 19876042
references.citation: Neurotherapeutics. 2010 Oct;7(4):392-8
references.pmid: 20880503
references.citation: J Neuroimmune Pharmacol. 2007 Mar;2(1):93-6
references.pmid: 18040831
references.citation: Biochem Soc Trans. 2012 Feb;40(1):85-9
references.pmid: 22260670
references.citation: Exp Neurol. 2004 Aug;188(2):246-53
references.pmid: 15246824
references.citation: Toxicol Sci. 2010 Nov;118(1):266-75
references.pmid: 20810542