a-85380 and Disease-Models--Animal

When Zika virus emerged as a public health emergency there were no drugs or vaccines approved for its prevention or treatment. We used a high-throughput screen for Zika virus protease inhibitors to identify several inhibitors of Zika virus infection. We expressed the NS2B-NS3 Zika virus protease and conducted a biochemical screen for small-molecule inhibitors. A quantitative structure-activity relationship model was employed to virtually screen ∼138,000 compounds, which increased the identification of active compounds, while decreasing screening time and resources. Candidate inhibitors were validated in several viral infection assays. Small molecules with favorable clinical profiles, especially the five-lipoxygenase-activating protein inhibitor, MK-591, inhibited the Zika virus protease and infection in neural stem cells. Members of the tetracycline family of antibiotics were more potent inhibitors of Zika virus infection than the protease, suggesting they may have multiple mechanisms of action. The most potent tetracycline, methacycline, reduced the amount of Zika virus present in the brain and the severity of Zika virus-induced motor deficits in an immunocompetent mouse model. As Food and Drug Administration-approved drugs, the tetracyclines could be quickly translated to the clinic. The compounds identified through our screening paradigm have the potential to be used as prophylactics for patients traveling to endemic regions or for the treatment of the neurological complications of Zika virus infection.

Topics: Animals; Antiviral Agents; Artificial Intelligence; Chlorocebus aethiops; Disease Models, Animal; Drug Evaluation, Preclinical; High-Throughput Screening Assays; Immunocompetence; Inhibitory Concentration 50; Methacycline; Mice, Inbred C57BL; Protease Inhibitors; Quantitative Structure-Activity Relationship; Small Molecule Libraries; Vero Cells; Zika Virus; Zika Virus Infection

Enhancement of cholinergic function via nicotinic acetylcholine (ACh) receptor (nAChR) agonism is a potential approach for the treatment of cognitive impairment associated with schizophrenia (CIAS). Some atypical antipsychotic drugs (AAPDs) enhance ACh release in rodent brain, indirectly stimulating these receptors. Here, we elucidate which nAChR subtypes mediate novel object recognition (NOR) in normal rats and contribute to the ability of the AAPD, lurasidone, to improve the NOR deficit in sub-chronic (sc) phencyclidine (PCP)-treated rats, a model for CIAS.. The ability of lurasidone and nAChR ligands to reverse the scPCP-induced deficit in NOR was assessed in female, Long-Evans rats.. The broad acting nAChR antagonist, mecamylamine (MEC), induced a NOR deficit in normal rats. The NOR deficit secondary to scPCP was reversed by either selective α4β2* nAChR agonism (A-85380) or α7 nAChRs agonism (PNU-282987); these effects were blocked by DHβE and MLA, selective antagonists of α4β2* and α7 nAChR, respectively. The ability of lurasidone to reverse the scPCP-induced NOR deficit was blocked by MEC, but not MLA or DHβE. However, sub-effective doses (SED) of either A-85380 or PNU-282987 potentiated the ability of SED lurasidone to reverse the scPCP-induced NOR deficit.. These results identify both α4β2* and α7 nAChRs as candidates for enhancing the ability of lurasidone and other AAPDs, which increase the release of ACh, to improve CIAS.

Topics: alpha7 Nicotinic Acetylcholine Receptor; Animals; Azetidines; Benzamides; Bridged Bicyclo Compounds; Cognition Disorders; Disease Models, Animal; Female; Lurasidone Hydrochloride; Mecamylamine; Nicotinic Agonists; Nicotinic Antagonists; Nootropic Agents; Phencyclidine; Random Allocation; Rats, Long-Evans; Receptors, Nicotinic

L-dopa therapy for Parkinson's disease leads to dyskinesias or abnormal involuntary movement (AIMs) for which there are few treatment options. Our previous data showed that nicotine administration reduced L-dopa-induced AIMs in parkinsonian monkeys and rats. To further understand how nicotine mediates its antidyskinetic action, we investigated the effect of nicotinic receptor (nAChR) agonists in unilateral 6-OHDA-lesioned rats with varying striatal damage. We first tested the drugs in L-dopa-treated rats with a near-complete striatal dopamine lesion (>99%), the standard rodent dyskinesia model. Varenicline, an agonist that interacts with multiple nAChRs, did not significantly reduce L-dopa-induced AIMs, while 5-iodo-A-85380 (A-85380), which acts selectively at α4β2* and α6β2* subtypes, reduced AIMs by 20%. By contrast, both varenicline and A-85380 reduced L-dopa-induced AIMs by 40-50% in rats with a partial striatal dopamine lesion. Neither drug worsened the antiparkinsonian action of L-dopa. The results show that selective nicotinic agonists reduce dyskinesias, and that they are optimally effective in animals with partial striatal dopamine damage. These findings suggest that presynaptic dopamine terminal α4β2* and α6β2* nAChRs are critical for nicotine's antidyskinetic action. The current data have important implications for the use of nicotinic receptor-directed drugs for L-dopa-induced dyskinesias, a debilitating motor complication of dopamine replacement therapy for Parkinson's disease.

Topics: Animals; Antiparkinson Agents; Azetidines; Benzazepines; Corpus Striatum; Disease Models, Animal; Dopamine Plasma Membrane Transport Proteins; Drug Therapy, Combination; Dyskinesia, Drug-Induced; Levodopa; Male; Nicotinic Agonists; Oxidopamine; Parkinson Disease; Quinoxalines; Rats; Rats, Sprague-Dawley; Varenicline

Recent evidence has suggested that the anti-allodynic effect of neuronal acetylcholine receptor (nAChR) agonists may have a peripheral component [L.E. Rueter, K.L. Kohlhaas, P. Curzon, C.S. Surowy, M.D. Meyer, Peripheral and central sites of action for A-85380 in the spinal nerve ligation model of neuropathic pain, Pain 103 (2003) 269-276]. In further studies of the peripheral anti-allodynic mechanisms of nAChR agonists, we investigated the function of nAChRs in acutely isolated dorsal root ganglion (DRG) neurons from allodynic [L5-L6 spinal nerve ligation (SNL)] and naive adult rats. Following determination of cell diameter and membrane capacitance, responses to rapid applications of nAChR agonists were recorded under whole cell patch clamp. nAChR inward currents were observed in approximately 60% of naive neurons, across small, medium, and large diameter cells. Evoked nAChR currents could be clustered into three broad classes: fast transient, biphasic, and slow desensitizing currents, consistent with multiple subtypes of nAChR expressed in DRG [J.R. Genzen, W. Van Cleve, D.S. McGehee, Dorsal root ganglion neurons express multiple nicotinic acetylcholine receptor subtypes, J. Neurophysiol. 86 (2001) 1773-1782]. In contrast, in neurons from allodynic animals, the occurrence and amplitude of responses to nAChR agonists were significantly reduced. Reduced responsiveness to nAChR agonists covered the range of DRG neuron sizes. The decrease in the responsiveness to nAChR agonists was not seen in neighboring uninjured L4 neurons. The significant decrease in the number of cells with nAChR agonist responses, compounded with the significant decrease in response amplitude, indicates that there is a marked down regulation of functional nAChRs in DRG somata associated with SNL.

Topics: Acetylcholine; Animals; Azetidines; Cell Count; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Interactions; Ganglia, Spinal; Ligation; Lumbosacral Region; Male; Mecamylamine; Membrane Potentials; Neurons; Nicotinic Agonists; Nicotinic Antagonists; Pain; Patch-Clamp Techniques; Rats; Rats, Sprague-Dawley; Receptors, Nicotinic; Spinal Cord Injuries

Evidence suggests that nicotinic receptors play a role in nigrostriatal function, a finding that may be relevant to Parkinson's disease. Knowledge of the conditions that regulate nicotinic receptor expression is therefore important. Previous studies showed that several different nicotinic receptors, including alpha-conotoxinMII (alpha-CtxMII)-sensitive receptors, are decreased after nigrostriatal damage. Nigrostriatal dopaminergic terminals also demonstrate a capacity for recovery after lesioning. The present experiments were therefore done to determine whether there were changes in striatal nicotinic receptors with recovery. To address this, we used two well-characterized animal models of nigrostriatal damage produced using the selective dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Studies in mice showed that striatal 125I-alpha-CtxMII, as well as 125I-epibatidine and 125I-A85380 binding sites significantly recovered 1 month after lesioning, suggesting that alpha6* and most likely alpha4* receptors are increased. Experiments were next done in monkeys since striatal 125I-alpha-CtxMII receptors constitute a large percentage of nicotinic receptors and are more vulnerable to nigrostriatal damage in this model that closely mirrors Parkinson's disease. In monkeys allowed to recover from the toxic effects of MPTP for a 1-2 year period, there was a significant improvement in the Parkinson disability score. There was also a reversal in lesion-induced declines in striatal alpha-CtxMII-sensitive receptors, but no significant change in 125I-epibatidine and 125I-A85380 receptors. These findings suggest that alpha3*/alpha6* sites are selectively increased in monkey striatum with recovery. The present data show that recovery of 125I-alpha-CtxMII receptors occurs in parallel with the dopamine transporter, indicating that these nicotinic receptors sites are localized to presynaptic dopamine terminals in both species.

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Animals; Azetidines; Bridged Bicyclo Compounds, Heterocyclic; Conotoxins; Corpus Striatum; Disease Models, Animal; Dopamine Plasma Membrane Transport Proteins; Female; Iodine Radioisotopes; Male; Membrane Glycoproteins; Membrane Transport Proteins; Mice; Mice, Inbred C57BL; Nerve Tissue Proteins; Neural Pathways; Parkinsonian Disorders; Presynaptic Terminals; Pyridines; Receptors, Nicotinic; Recovery of Function; Saimiri; Substantia Nigra; Up-Regulation