Development of a novel therapeutic suppressor of brain proinflammatory cytokine up-regulation that attenuates synaptic dysfunction and behavioral deficits

Abstract

We report the development of a novel, aqueous-soluble, safe, small molecule, experimental therapeutic that suppresses injury-induced, proinflammatory cytokine increases in the brain, with resultant attenuation of synaptic protein biomarker loss and improvement in hippocampus-dependent behavioral deficits. A GMP production scheme for the active pharmaceutical ingredient, compound 17, is presented. The development and large-scale availability of this novel compound allow exploration of new, potentially disease-modifying, therapeutic approaches to CNS disorders.Over-production of proinflammatory cytokines contributes to the progression of pathology in a diverse array of diseases, 1-5 and the recent approval of protein therapies 5-8 that seek to control the level or activity of proinflammatory cytokines provides a proof-of-concept for targeting proinflammatory cytokine levels in drug discovery. However, the macromolecular therapeutic approach is not as amenable to clinical use in some diseases, such as central nervous system (CNS) disorders, creating an unmet need for small molecule, brain-penetrant compounds as new classes of disease-modifying drugs. An accumulating body of evidence from animal model studies and clinical observations supports the concept 1,3,4,9,10 that development of such therapies for CNS disorders could have potential impact on both acute and chronic disorders as diverse as Alzheimer's disease (AD), stroke, traumatic brain injury, neuropathic pain, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, agerelated macular degeneration, and non-AD dementias. The prevailing hypothesis in this regard is that targeting the up-regulation of proinflammatory cytokine production by activated glia, the main cellular source of cytokines in the CNS, will alter disease progression through attenuation of the subsequent neuronal synaptic dysfunction, the cellular basis of clinical symptoms and behavioral alterations. However, there are no such current therapies available and there are no consensus molecular targets for such discovery efforts. The current state of the art requires, therefore, the use of the more classical and unbiased functional approach to drug discovery. 11 *To Whom Correspondence Should be Addressed: Phone: 312-503-0656, FAX: 312-503-0007, E-mail: m-watterson@northwestern.edu. SUPPORTING INFORMATION AVAILABLE:The contents of Supporting Information include the following: details of synthetic procedures, analytical chemistry methods, physical property determinations, NMR spectra, and biological procedures, including screening data for oral bioavailability.Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affec...

Molecular Properties and CYP2D6 Substrates: Central Nervous System Therapeutics Case Study and Pattern Analysis of a Substrate Database

“…Analytical Chemistry. All the HPLC analytical data were obtained on a commercially available system from Dionex Corp. (Sunnyvale, CA) by using a Phenomenex (Torrance, CA) Luna C18 column (250 ϫ 2.0 mm; 5 m) with guard column as described previously (Hu et al, 2007). Peak quantification was done based on absorption measurements at 260 nm relative to a standard curve obtained by serial dilutions of compounds.…”

“…Minozac, MW01-5-079HAB, and MW01-5-042HAB were synthesized and characterized as described previously (Hu et al, 2007). Synthetic reaction progress was monitored by HPLC, and final products were characterized by mass spectrometry, HPLC, and 1 H NMR as described previously (Hu et al, 2007).…”

“…In this regard, minaprine and minozac provide a novel study of structurally similar compounds (Tanimoto similarity coefficient ϭ 0.71) with distinct pharmacology and CNS activity that are also different in their CYP2D6 substrate status. In terms of prevailing models, the difference in CYP2D6 status of minaprine and minozac cannot be explained by differences in lipophilicity (de Groot et al, 1997) because the compounds possess nearly identical experimental log 10 of the octanol/ water partition coefficient (LogP) values of 1.9 (Arnaud-Neu et al, 1990) and 2.3 (Hu et al, 2007), respectively. Although minaprine adheres to the prevailing localized charge model for CYP2D6 substrate status (de Groot et al, 1997), in that it is more basic than the nonsubstrate minozac, and a quantitative model of CYP2D6 substrates provides an accurate forecast for minaprine (Haji-Momenian et al, 2003), the quantitative structure-activity relationship model cannot explain why minozac is not a substrate.…”

Minozac Treatment Prevents Increased Seizure Susceptibility in a Mouse “Two-Hit” Model of Closed Skull Traumatic Brain Injury and Electroconvulsive Shock-Induced Seizures

“…To determine whether suppression of the acute increase in proinflammatory cytokines after TBI would attenuate the subsequent increase in susceptibility to ECS-induced seizures, we used Minozac (Mzc), a small molecule, selective inhibitor of proinflammatory cytokine production by activated glia (Hu et al, 2007;Lloyd et al, 2008). We used the same dose and route of administration (5 mg/kg IP injection), and similar timing of administration (a total of 2 doses of Mzc, given at 3 h and 6 h following TBI), to those used in previous studies of TBI (Lloyd et al, 2008), and a two-hit model of seizures (Somera-Molina et al, 2007.…”

Pharmacogenomics of Alzheimer’s disease and cerebral palsy: implications of the apolipoprotein E genotype

“…Activated microglia contribute to neuronal injury by the production of multiple factors, including cytokines, nitric oxide and reactive oxygen species [30,31]. Both oxidative stress [32] and the glial activation cycle [33,34] are promising therapeutic targets in AD.…”