No abstract

Recent studies indicate that the time required for virus‐infected cells to become vulnerable for the activity of CTL is of significance for the capacity of CTL to control ongoing viral reproduction. To investigate whether this applies to the effectiveness of HIV‐1‐specific CTL, we measured virus production in cultures containing CD4+ T cells inoculated with HIV at low multiplicity of infection, and CTL directed against an early protein, Rev, or a late protein, RT. The Rev‐specific CTL prevented at least 2 log10 more HIV‐1 production, in 10 days, than similar numbers of RT‐specific CTL. To study how CTL effectiveness depends on variations in the potency of effector functions and kinetics of HIV protein expression, we developed a mathematical model describing CTL‐target cell interactions during successive infection cycles. The results show that substantially higher CTL‐mediated target cell elimination rates are required to achieve control as there is less time for CTL to act before infected cells release progeny virions. Furthermore, in vitro experiments with HIV recombinant viruses showed that the RT‐specific CTL were at least as effective as the Rev‐specific CTL, but only if the RT epitope was expressed as part of the early protein Nef. Together these results indicate that CTL control ongoing HIV reproduction more effectively if they are able to recognize infected cells earlier during individual viral replication cycles. This provides rationale for immunization strategies that aim at inducing, boosting or skewing CTL responses to early regulatory proteins in AIDS vaccine development.

The design and study of two classes of noncompetitive acetylcholinesterase inhibitors (AChEIs) which also function as NSAID prodrugs are reported. The most potent AChEIs disclosed contain an aromatic alkyl-aryl linker between an NSAID and a lipophilic choline mimic and they inhibit acetylcholinesterase (AChE) in the submicromolar range. These agents have the therapeutic potential to dually target inflammation by releasing an NSAID in vivo and activating the cholinergic antiinflammatory pathway via cholinergic up-regulation.Pro-inflammatory cytokine up-regulation plays a role in the pathogenesis of a wide range of disorders including osteoarthritis, 1 psoriasis, 2 multiple sclerosis, 3 and other autoimmune disorders. 4 Despite decades of research, non-steroidal anti-inflammatory drugs (NSAIDs) are still one of the most commonly used, highly effective treatments for such disorders. However, since chronic NSAID-use often leads to gastrointestinal (GI) side effects, NSAID ester prodrugs have been explored to mask the acidic GI-irritating portion of the NSAID. 5 For conditions such as arthritis, topical NSAID prodrugs are particularly useful as their therapeutic action is localized, resulting in minimal systemic side effects. 6 Alternately, acetylcholinesterase inhibitors (AChEIs) have implications in the treatment of severe inflammation resulting from sepsis, 7 endotoxemia, 8 and rheumatoid arthritis, 9 as well as in the treatment of neuroinflammation associated with Alzheimer's disease [10][11][12] and Myasthenia Gravis. 13 The administration of CNS-active AChEIs such as galanthamine depletes systemic pro-inflammatory cytokines and ameliorates both central and peripheral inflammation. 8 AChEIs seem to suppress inflammation via the cholinergic anti-inflammatory pathway, a mechanism by which the vagus nerve of the CNS regulates the production and release of tumor necrosis factor and other cytokines. 14,15 *Corresponding author. Tel.: 1-610-758-3464; fax.: 1-610-758-3461; ndh0@lehigh.edu. † Current address: PTC Therapeutics, Inc., South Plainfield, NJ 07080-2449, USA ‡ Current address: Digestive Care, Inc., Bethlehem, PA 18017-7059, USA Supplementary data Synthetic procedures, physical characterization of compounds, in vitro assay methods and Lineweaver-Burk plots can be found in the online version of this article.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 affect the content, and all legal disclaimers that apply to the journal pertain. (Figure 1). 18 The synthesis and evaluation of these compounds as anti-inflammatory and anticholinesterase agents for topical or oral administration are presented herein. NIH Public AccessAuthor Manuscript Bioor...

The azetidinone LY307174 (1) was identified as a screening lead for the vasopressin V1a receptor (IC 50 45 nM at the human V1a receptor) based on molecular similarity to ketoconazole (2), a known antagonist of the luteinizing hormone releasing hormone receptor. Structure-activity relationships for the series were explored to optimize receptor affinity and pharmacokinetic properties, resulting in compounds with K i values < 1 nM and brain levels after oral dosing ~100-fold higher than receptor affinities.The neurohypophysial hormones vasopressin 1 and oxytocin exert a wide range of physiological effects through binding to specific membrane receptors belonging to the G protein-coupled receptor (GPCR) superfamily. To date, three vasopressin receptor subtypes and one oxytocin receptor have been pharmacologically and functionally described 1 . V1a, V1b, and oxytocin receptors activate phospholipase C, resulting in the production of inositol 1,4,5-trisphosphate and diacylglycerol, mobilization of intracellular calcium, and activation of protein kinase C. V2 receptors stimulate adenylyl cyclase, resulting in the accumulation of cyclic AMP and activation of protein kinase A. All four receptor subtypes from several mammalian species have been recently cloned 2-5 , as well as closely related receptors from bony fishes and invertebrates 6, 7 . Although vasopressin is perhaps best-known for its role in the cardiovascular system, it also has actions in the central nervous system (CNS), and several CNS applications of vasopressin receptor antagonists have been suggested (reviewed in references 8 and 9 ). A number of research groups have prepared antagonists directed at the vasopressin V1 receptor 10-15 . While V1a antagonists have been made, none of these have been reported to penetrate the CNS efficiently. 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 affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptBioorg Med Chem. Author manuscript; available in PMC 2007 November 8. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptOur vasopressin antagonist program was initiated to identify a CNS-active V1a antagonist: one with potent affinity for the human V1a receptor (IC 50 < 10 nM), good oral availability, and ability to penetrate the blood brain barrier -in short, a candidate for human clinical development targeting CNS disorders. The program began at Lilly in 1990 with the selection of a 1,500-compound "Neuropeptide Cassette" -a library intended to identify nonpeptide ligands for neuropeptide receptors. The library applied the concept of receptor crosstalk -previously well...