ABSTRACT
A HTS campaign identified compound 1, an excellent hit-like molecule to initiate medicinal chemistry efforts to optimize a dual ROCK1 and ROCK2 inhibitor. Substitution (2-Cl, 2-NH2, 2-F, 3-F) of the pyridine hinge binding motif or replacement with pyrimidine afforded compounds with a clean CYP inhibition profile. Cocrystal structures of an early lead compound were obtained in PKA, ROCK1, and ROCK2. This provided critical structural information for medicinal chemistry to drive compound design. The structural data indicated the preferred configuration at the central benzylic carbon would be ( R), and application of this information to compound design resulted in compound 16. This compound was shown to be a potent and selective dual ROCK inhibitor in both enzyme and cell assays and efficacious in the retinal nerve fiber layer model after oral dosing. This tool compound has been made available through the AbbVie Compound Toolbox. Finally, the cocrystal structures also identified that aspartic acid residues 176 and 218 in ROCK2, which are glutamic acids in PKA, could be targeted as residues to drive both potency and kinome selectivity. Introduction of a piperidin-3-ylmethanamine group to the compound series resulted in compound 58, a potent and selective dual ROCK inhibitor with excellent predicted drug-like properties.
Subject(s)
Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/pharmacology , rho-Associated Kinases/antagonists & inhibitors , Administration, Oral , Animals , Biological Availability , Crystallography, X-Ray , Cytochrome P-450 CYP2C9 Inhibitors/chemistry , Cytochrome P-450 CYP2C9 Inhibitors/pharmacology , Cytochrome P-450 CYP3A Inhibitors/chemistry , Cytochrome P-450 CYP3A Inhibitors/pharmacology , Drug Design , Drug Evaluation, Preclinical/methods , Half-Life , Humans , Mice, Inbred C57BL , Optic Nerve Injuries/drug therapy , Optic Nerve Injuries/pathology , Rats, Sprague-Dawley , Structure-Activity Relationship , rho-Associated Kinases/chemistryABSTRACT
The individual isomers of methyl 1-amino-3-(4-bromophenyl)cyclopentanecarboxylate are useful intermediates for the synthesis of S1P1 receptor agonists. Herein we describe a scalable synthesis and isolation of each of the four stereoisomers of this compound in gram quantities with >98% ee and de. The utility of this approach is demonstrated by the synthesis of ((1R,3R)-1-amino-3-(4-octylphenyl)cyclopentyl)methanol in 7 steps, 11% overall yield, and >98% ee and de.
Subject(s)
Carboxylic Acids/chemical synthesis , Cyclopentanes/chemical synthesis , Receptors, Lysosphingolipid/antagonists & inhibitors , Carboxylic Acids/chemistry , Cyclopentanes/chemistry , Molecular Structure , Receptors, Lysosphingolipid/metabolism , Stereoisomerism , Structure-Activity RelationshipABSTRACT
Automated semipreparative LC/MS systems are now well established commercially and commonly used for purification of early stage drug discovery compounds. A number of vendors have instruments on the market that are capable of reliably purifying compounds with good water/acetonitrile solubility. However, these systems often fail when the sample has poor solubility, extreme polarity, and/or poor ionization. Even in cases when substantial optimization has been done prior to purification, a certain percent of failures to recover the desired product is unavoidable. In the past, when the majority of samples run on LC/MS semipreparative systems were large combinatorial libraries, some losses in this high throughput mode were acceptable. However, now that more chemistry laboratories are making smaller more focused libraries with higher purity requirements, reliability and recovery are more crucial. This paper describes modifications made to customize an MS-triggered semipreparative LC/MS system in order to ensure improved reliability and recovery of products from traditional medicinal chemistry as well as combinatorial libraries.
Subject(s)
Mass Spectrometry/instrumentation , Chromatography, Liquid/instrumentation , Combinatorial Chemistry Techniques , Equipment Design , Small Molecule Libraries/chemistryABSTRACT
Compound 1 (FTY720, Fingolimod) represents a new generation of immunosuppressant that modulates lymphocyte trafficking by interacting with the S1P(1) receptor. Compound 1 also provides a template molecule for studying the molecular biology of S1P receptors and related enzymes (kinases and phosphatases). In this study, two conformationally constrained analogues of 1 ( 3a and 3c) were asymmetrically synthesized in high optical purity. In vitro assessment documented that both analogues are Sphk2 substrates, their phosphorylated species are potent S1P(1) receptor agonists, and 3a-P is a potent S1P 3 antagonist. After oral administration in mice, both compounds evoked lymphopenia, but their duration of action differed markedly.
