- Email: [email protected]
PS1DE9 MICE
P1018 P4-009
Poster Presentations: Wednesday, July 27, 2016 DESIGN, SYNTHESIS AND MOLECULAR DOCKING STUDIES OF SOME NOVEL POTENTIAL PIPERAZINE DERIVATIVES FOR THE IMPROVEMENT OF COGNITIVE DYSFUNCTION
Poonam Piplani1, Chhanda Charan Danta2, 1Panjab University, Chandigarh, India; 2Panjab University, Chandigarh, India. Contact e-mail: [email protected] Background: Cognitive dysfunction (CD) is an impairment of the
brain’s functions that are related to memory, attention, intellect etc. It also includes aging associated cognitive decline,1 mild cognitive decline2 and cognitive impairment no dementia (CIND). Cognitive dysfunction has become a major public health concern because of its increasing prevalence and financial costs of care.3 Methods: Series of novel piperazine derivatives have been designed, synthesized and evaluated for cognition enhancing activity. The synthesized compounds were screened for their in vitro AChE inhibition and reversal of scopolamine induced memory deficit in a passive avoidance stepdown animal model in mice. ADME properties were predicted using cheminformatics tools. The antiradical activity was evaluated by DPPH method. Enzyme kinetics and molecular docking studies were carried out to elucidate the mechanism of AChE inhibition. Results: All the compounds exhibited excellent IC50 values with potential dual binding site inhibition activity. The IC50 values and inhibition constants of the most promising compounds 1d and 3c were found to be 2.23 mM, 1.05 mM, 14.38 mM and 6.93 mM respectively. They potentially reversed the scopolamine induced memory deficit at a dose of 1.0 mg/kg i.p. in mice. Furthermore, 1d and 3c showed high CNS penetration and brain AChE inhibition in ex vivo experiments in mice. Additionally, these compounds also showed free radical scavenging activity. Conclusions: The present study focused on the development of piperazine based new chemical entities that can be well received for the treatment of cognitive dysfunction manifested in various neurological disorders. The study put forward two lead molecules 1d and 3c that exhibited appreciable acetylcholinesterase inhibitory potential. Further elaborations in this domain of research can bring out novel compounds that can revolutionize the paradigm of treatment of neurodegenerative disorders.
the extracellular increase of the NMDA receptor co-agonist glycine by glycine transporter-1 (GlyT1) inhibition. Thus, inhibiting GlyT1 has potential for treating memory impairment in Alzheimer’s disease via strengthening glutamatergic neurotransmission. Previous studies with GlyT1-inhibitors show memory-enhancing effects in animals. This study characterizes the potency and selectivity of the novel GlyT1 inhibitor BI 425809 and its effects on glycine increase in rat cerebrospinal fluid (CSF). BI 425809 was also evaluated in two rodent cognition tasks assessing episodic memory and working memory. Methods: The molecular potency of BI 425809 for GlyT1 was determined by inhibition of [3H]-glycine uptake in human SK-N-MC cells and rat primary cortical neurons. Selectivity was tested against GlyT2 and other off-targets by uptake and receptor binding assays, respectively. Concentrations of glycine in rat CSF samples collected from cisterna magna following oral administration of BI 425809 were determined by HPLC-MS/MS technique. BI 425809 was tested after oral application in the mouse T-maze spontaneous alternation test and in the social recognition test in naive rats. Results: The IC50 value of BI 425809 on GlyT1 was 4.0 nM in SK-N-MC cells and 4.4 nM in rat primary neurons. BI 425809 demonstrated no relevant activity against GlyT2 (IC50 >10 mM) and 103 off-targets at 10 mM. A dose-dependent increase of glycine in rat CSF was observed. BI 425809 reversed MK-801induced memory deficits in the mouse T-maze (working memory) and improved memory performance in the rat social recognition task (episodic memory). Conclusions: BI 425809 is a potent and selective GlyT1 inhibitor. Systemic administration of BI 425809 led to an increase in glycine levels in rat CSF demonstrating functional target engagement – i.e. GlyT1 inhibition in the brain – showing that glycine levels in CSF can be used to assess GlyT1 inhibition centrally. This might also be used to evaluate central target engagement in clinical trials. BI 425809 showed memory enhancing effects in animal cognition tests. GlyT1 inhibition may be a potential approach to pharmacologically improve cognition in Alzheimer’s disease.
P4-011
INTRANASAL INSULIN ALLEVIATES INSULIN SIGNALING DEFICITS AND AMYLOID PATHOLOGY IN YOUNG ADULT APPSWE/PS1DE9 MICE
Yanxing Chen, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. Contact e-mail: chenyanxing84@ gmail.com Background: Bain insulin signaling deficits have been demonstrated
P4-010
IMPROVING COGNITIVE FUNCTION IN RODENTS VIA INCREASING GLYCINE LEVELS IN BRAIN BY THE NOVEL GLYCINE TRANSPORTER1 INHIBITOR BI 425809
Holger Rosenbrock1, Riccardo Giovannini1, Bernhard Schmid1, Gert Kramer1, Roberto Arban1, Cornelia Dorner-Ciossek1, Glen Wunderlich2, 1Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany; 2Boehringer Ingelheim (Canada) Ltd, Burlington, ON, Canada. Contact e-mail: glen.wunderlich@ boehringer-ingelheim.com Background: Dysfunction of the glutamatergic system related to the NMDA receptor may play an important role in cognitive impairment in Alzheimer’s disease. One approach to counteract this is
to contribute to multiple major pathological features in Alzheimer’s disease (AD). Although intranasal insulin has shown efficacy in AD patients, little is known about the underlying mechanisms. Methods: We investigated the potential disease-modifying effects of intranasal insulin on young adult APPswe/PS1dE9 mice at the early stage of AD pathologies. We treated 4.5-month old APPswe/ PS1dE9 mice with intransal insulin for 6 weeks, and investigated the cognition and amyloid pathology in these mice. Results: we found that intranasal insulin improved behavioral flexibility, enhanced defective brain insulin signaling, and reduced b-amyloid (Ab) production and plaque formation in APPswe/PS1dE9 mice. Furthermore, c-Jun N-terminal kinase (JNK) activation, which plays a pivotal role in insulin resistance and AD pathologies, was significantly decreased by intranasal insulin. We also found that alleviated amyloid pathology by intranasal insulin was resulted
Poster Presentations: Wednesday, July 27, 2016
from enhanced nonamyloidogenic processing and compromised amyloidogenic processing of amyloid precursor protein (APP), as well as a reduction in aggregation and fibril formation of Ab. In addition, intranasal insulin also promoted neurogenesis in APPswe/PS1dE9 mice. Conclusions: The present study unveils the mechanisms underlying the beneficial effects of intranasal insulin on Ab pathologies in vivo for the first time. Our results support that intranasal insulin can be a very promosing therapeutics for the prevention and treatment of AD.
days 1 and 3 (3 mg/kg) and Insulin (3 or 6mU/icv) or saline injection was started from day 4 and continued in an everyday manner till day 13. The animal’s learning and memory capability was assessed on days 15-18 using Morris water maze. Results: The results showed that insulin protects animals in a dose dependent manner. In dose 3 mU it was more effective in preventing memory loss. Conclusions: This study confirms the beneficial effect of exogenous insulin against memory loss and suggests its usage consideration in clinic.
P4-013 P4-012
INTRAVENTRICULAR INSULIN TREATMENT PREVENTS AGAINST CENTRAL STZ-INDUCED IMPAIRMENT IN SPATIAL LEARNING AND MEMORY
Maryam Moosavi1, Abbas Bahramian2, Karim Rastegar3, 1Shiraz University Medical Science, Shiraz, Iran; 2Shiraz University of Medical Sciences, Shiraz, Iran (Islamic Republic of); 3Shiraz University of Medical Sciences, Shiraz, Iran (Islamic Republic of). Contact e-mail: marmoosavi@ sums.ac.ir Background: Insulin has been shown to have a role in pathology of
Alzheimer’s disease (AD) in a way that sometimes AD is referred as diabetes type 3. Centrally administered streptozotocin (STZ) is known to induce an animal model of memory impairment in rodents. In order to know more the effect of insulin in learning and memory process, this study assessed if insulin (icv) is able to prevent central STZ-induced amnesia. Methods: Adult male SpragueDawely rats weighing 200-250 g were used. The canules were implanted bilaterally into lateral ventricles. STZ was administered on
P1019
PRO-COGNITIVE AND ANTI-INFLAMMATORY EFFECTS OF AF710B, A MIXED M1 MUSCARINIC/ SIGMA-1 RECEPTOR AGONIST, IN THE MCGILLR-THY1-APP RAT MODEL OF HUMAN AD-LIKE AMYLOID PATHOLOGY
Helene Hall1, Maria Florencia Iulita1, Adriana Ducatenzeiler1, Abraham Fisher2, A. Claudio Cuello1, 1McGill University, Montreal, QC, Canada; 2Israel Institute for Biological Research, Ness Ziona, Israel. Contact e-mail: [email protected] Background: Treatments currently available for Alzheimer’s disease (AD) are merely symptomatic and initiated at a stage where damage to the brain is irreversible, highlighting the pressing need for disease-modifying drugs that could potentially halt or reverse progression of the pathology. AF710B is a novel concomitant agonist of the M1 muscarinic and sigma-1 receptors, which rescues synaptic loss and reduces oxidative stress and apoptosis in vitro. In 3xTg-AD mice, AF710B attenuates cognitive deficits and decreases ADlike hallmarks (such as BACE1, GSK3beta activity, p25/CDK5, neuro-inflammation, Amyloid-beta, plaques and tau pathologies) [1]. In a further extension, we tested whether AF710B can reduce AD-like neuropathology and alleviate cognitive impairment in McGill-R-Thy1-APP transgenic (tg) rats. Methods: AF710B (10mg/kg) or vehicle (PBS) was administered orally to tg versus wild type (wt) rats of 12 months of age, daily for 4.5 months. Treatment was interrupted for 5 weeks before submitting rats to behavioral tests (Novel Object Recognition, Morris Water Maze), a unique design to test AF710B true disease-modifying properties. Post-mortem brain tissue was submitted to biochemical (ELISA for Amyloid-beta load) and histological (microglia counts) analyses. Results: Long term chronic treatment with AF710B was capable of fully reverting the cognitive deficits present at later stages of the AD-like Amyloid-beta neuropathology in the McGill-R-Thy1-APP tg rats, as evidenced by a complete reversal of the deficit measured by the Novel Object Recognition and Morris Water Maze tasks. Importantly, this effect was maintained following a 5-week interruption in the treatment, suggesting true disease-modifying properties of AF710B. An ELISA performed on cortical samples showed a trend towards a decrease in Amyloid-beta 42 soluble and insoluble fractions in treated tg rats, supporting a possible shift in the APP processing towards a nonamyloidogenic pathway. McGill-R-Thy1-APP rats display an increase in the number of microglia recruited towards CA1 hippocampal Amyloid-beta burdened-neurons; AF710B treatment normalized the number of microglial cells to that of untreated wt rats, pointing to anti-inflammatory properties. Conclusions: With M1/sigma-1 activity as well as putative disease-modifying properties at very low doses, AF710B is a promising novel therapeutic agent for the treatment of AD. [1] Fisher et al, 2015 Neurodegener Dis. DOI:10.1159/000440864.
Poster Presentations: Wednesday, July 27, 2016 DESIGN, SYNTHESIS AND MOLECULAR DOCKING STUDIES OF SOME NOVEL POTENTIAL PIPERAZINE DERIVATIVES FOR THE IMPROVEMENT OF COGNITIVE DYSFUNCTION
Poonam Piplani1, Chhanda Charan Danta2, 1Panjab University, Chandigarh, India; 2Panjab University, Chandigarh, India. Contact e-mail: [email protected] Background: Cognitive dysfunction (CD) is an impairment of the
brain’s functions that are related to memory, attention, intellect etc. It also includes aging associated cognitive decline,1 mild cognitive decline2 and cognitive impairment no dementia (CIND). Cognitive dysfunction has become a major public health concern because of its increasing prevalence and financial costs of care.3 Methods: Series of novel piperazine derivatives have been designed, synthesized and evaluated for cognition enhancing activity. The synthesized compounds were screened for their in vitro AChE inhibition and reversal of scopolamine induced memory deficit in a passive avoidance stepdown animal model in mice. ADME properties were predicted using cheminformatics tools. The antiradical activity was evaluated by DPPH method. Enzyme kinetics and molecular docking studies were carried out to elucidate the mechanism of AChE inhibition. Results: All the compounds exhibited excellent IC50 values with potential dual binding site inhibition activity. The IC50 values and inhibition constants of the most promising compounds 1d and 3c were found to be 2.23 mM, 1.05 mM, 14.38 mM and 6.93 mM respectively. They potentially reversed the scopolamine induced memory deficit at a dose of 1.0 mg/kg i.p. in mice. Furthermore, 1d and 3c showed high CNS penetration and brain AChE inhibition in ex vivo experiments in mice. Additionally, these compounds also showed free radical scavenging activity. Conclusions: The present study focused on the development of piperazine based new chemical entities that can be well received for the treatment of cognitive dysfunction manifested in various neurological disorders. The study put forward two lead molecules 1d and 3c that exhibited appreciable acetylcholinesterase inhibitory potential. Further elaborations in this domain of research can bring out novel compounds that can revolutionize the paradigm of treatment of neurodegenerative disorders.
the extracellular increase of the NMDA receptor co-agonist glycine by glycine transporter-1 (GlyT1) inhibition. Thus, inhibiting GlyT1 has potential for treating memory impairment in Alzheimer’s disease via strengthening glutamatergic neurotransmission. Previous studies with GlyT1-inhibitors show memory-enhancing effects in animals. This study characterizes the potency and selectivity of the novel GlyT1 inhibitor BI 425809 and its effects on glycine increase in rat cerebrospinal fluid (CSF). BI 425809 was also evaluated in two rodent cognition tasks assessing episodic memory and working memory. Methods: The molecular potency of BI 425809 for GlyT1 was determined by inhibition of [3H]-glycine uptake in human SK-N-MC cells and rat primary cortical neurons. Selectivity was tested against GlyT2 and other off-targets by uptake and receptor binding assays, respectively. Concentrations of glycine in rat CSF samples collected from cisterna magna following oral administration of BI 425809 were determined by HPLC-MS/MS technique. BI 425809 was tested after oral application in the mouse T-maze spontaneous alternation test and in the social recognition test in naive rats. Results: The IC50 value of BI 425809 on GlyT1 was 4.0 nM in SK-N-MC cells and 4.4 nM in rat primary neurons. BI 425809 demonstrated no relevant activity against GlyT2 (IC50 >10 mM) and 103 off-targets at 10 mM. A dose-dependent increase of glycine in rat CSF was observed. BI 425809 reversed MK-801induced memory deficits in the mouse T-maze (working memory) and improved memory performance in the rat social recognition task (episodic memory). Conclusions: BI 425809 is a potent and selective GlyT1 inhibitor. Systemic administration of BI 425809 led to an increase in glycine levels in rat CSF demonstrating functional target engagement – i.e. GlyT1 inhibition in the brain – showing that glycine levels in CSF can be used to assess GlyT1 inhibition centrally. This might also be used to evaluate central target engagement in clinical trials. BI 425809 showed memory enhancing effects in animal cognition tests. GlyT1 inhibition may be a potential approach to pharmacologically improve cognition in Alzheimer’s disease.
P4-011
INTRANASAL INSULIN ALLEVIATES INSULIN SIGNALING DEFICITS AND AMYLOID PATHOLOGY IN YOUNG ADULT APPSWE/PS1DE9 MICE
Yanxing Chen, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. Contact e-mail: chenyanxing84@ gmail.com Background: Bain insulin signaling deficits have been demonstrated
P4-010
IMPROVING COGNITIVE FUNCTION IN RODENTS VIA INCREASING GLYCINE LEVELS IN BRAIN BY THE NOVEL GLYCINE TRANSPORTER1 INHIBITOR BI 425809
Holger Rosenbrock1, Riccardo Giovannini1, Bernhard Schmid1, Gert Kramer1, Roberto Arban1, Cornelia Dorner-Ciossek1, Glen Wunderlich2, 1Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany; 2Boehringer Ingelheim (Canada) Ltd, Burlington, ON, Canada. Contact e-mail: glen.wunderlich@ boehringer-ingelheim.com Background: Dysfunction of the glutamatergic system related to the NMDA receptor may play an important role in cognitive impairment in Alzheimer’s disease. One approach to counteract this is
to contribute to multiple major pathological features in Alzheimer’s disease (AD). Although intranasal insulin has shown efficacy in AD patients, little is known about the underlying mechanisms. Methods: We investigated the potential disease-modifying effects of intranasal insulin on young adult APPswe/PS1dE9 mice at the early stage of AD pathologies. We treated 4.5-month old APPswe/ PS1dE9 mice with intransal insulin for 6 weeks, and investigated the cognition and amyloid pathology in these mice. Results: we found that intranasal insulin improved behavioral flexibility, enhanced defective brain insulin signaling, and reduced b-amyloid (Ab) production and plaque formation in APPswe/PS1dE9 mice. Furthermore, c-Jun N-terminal kinase (JNK) activation, which plays a pivotal role in insulin resistance and AD pathologies, was significantly decreased by intranasal insulin. We also found that alleviated amyloid pathology by intranasal insulin was resulted
Poster Presentations: Wednesday, July 27, 2016
from enhanced nonamyloidogenic processing and compromised amyloidogenic processing of amyloid precursor protein (APP), as well as a reduction in aggregation and fibril formation of Ab. In addition, intranasal insulin also promoted neurogenesis in APPswe/PS1dE9 mice. Conclusions: The present study unveils the mechanisms underlying the beneficial effects of intranasal insulin on Ab pathologies in vivo for the first time. Our results support that intranasal insulin can be a very promosing therapeutics for the prevention and treatment of AD.
days 1 and 3 (3 mg/kg) and Insulin (3 or 6mU/icv) or saline injection was started from day 4 and continued in an everyday manner till day 13. The animal’s learning and memory capability was assessed on days 15-18 using Morris water maze. Results: The results showed that insulin protects animals in a dose dependent manner. In dose 3 mU it was more effective in preventing memory loss. Conclusions: This study confirms the beneficial effect of exogenous insulin against memory loss and suggests its usage consideration in clinic.
P4-013 P4-012
INTRAVENTRICULAR INSULIN TREATMENT PREVENTS AGAINST CENTRAL STZ-INDUCED IMPAIRMENT IN SPATIAL LEARNING AND MEMORY
Maryam Moosavi1, Abbas Bahramian2, Karim Rastegar3, 1Shiraz University Medical Science, Shiraz, Iran; 2Shiraz University of Medical Sciences, Shiraz, Iran (Islamic Republic of); 3Shiraz University of Medical Sciences, Shiraz, Iran (Islamic Republic of). Contact e-mail: marmoosavi@ sums.ac.ir Background: Insulin has been shown to have a role in pathology of
Alzheimer’s disease (AD) in a way that sometimes AD is referred as diabetes type 3. Centrally administered streptozotocin (STZ) is known to induce an animal model of memory impairment in rodents. In order to know more the effect of insulin in learning and memory process, this study assessed if insulin (icv) is able to prevent central STZ-induced amnesia. Methods: Adult male SpragueDawely rats weighing 200-250 g were used. The canules were implanted bilaterally into lateral ventricles. STZ was administered on
P1019
PRO-COGNITIVE AND ANTI-INFLAMMATORY EFFECTS OF AF710B, A MIXED M1 MUSCARINIC/ SIGMA-1 RECEPTOR AGONIST, IN THE MCGILLR-THY1-APP RAT MODEL OF HUMAN AD-LIKE AMYLOID PATHOLOGY
Helene Hall1, Maria Florencia Iulita1, Adriana Ducatenzeiler1, Abraham Fisher2, A. Claudio Cuello1, 1McGill University, Montreal, QC, Canada; 2Israel Institute for Biological Research, Ness Ziona, Israel. Contact e-mail: [email protected] Background: Treatments currently available for Alzheimer’s disease (AD) are merely symptomatic and initiated at a stage where damage to the brain is irreversible, highlighting the pressing need for disease-modifying drugs that could potentially halt or reverse progression of the pathology. AF710B is a novel concomitant agonist of the M1 muscarinic and sigma-1 receptors, which rescues synaptic loss and reduces oxidative stress and apoptosis in vitro. In 3xTg-AD mice, AF710B attenuates cognitive deficits and decreases ADlike hallmarks (such as BACE1, GSK3beta activity, p25/CDK5, neuro-inflammation, Amyloid-beta, plaques and tau pathologies) [1]. In a further extension, we tested whether AF710B can reduce AD-like neuropathology and alleviate cognitive impairment in McGill-R-Thy1-APP transgenic (tg) rats. Methods: AF710B (10mg/kg) or vehicle (PBS) was administered orally to tg versus wild type (wt) rats of 12 months of age, daily for 4.5 months. Treatment was interrupted for 5 weeks before submitting rats to behavioral tests (Novel Object Recognition, Morris Water Maze), a unique design to test AF710B true disease-modifying properties. Post-mortem brain tissue was submitted to biochemical (ELISA for Amyloid-beta load) and histological (microglia counts) analyses. Results: Long term chronic treatment with AF710B was capable of fully reverting the cognitive deficits present at later stages of the AD-like Amyloid-beta neuropathology in the McGill-R-Thy1-APP tg rats, as evidenced by a complete reversal of the deficit measured by the Novel Object Recognition and Morris Water Maze tasks. Importantly, this effect was maintained following a 5-week interruption in the treatment, suggesting true disease-modifying properties of AF710B. An ELISA performed on cortical samples showed a trend towards a decrease in Amyloid-beta 42 soluble and insoluble fractions in treated tg rats, supporting a possible shift in the APP processing towards a nonamyloidogenic pathway. McGill-R-Thy1-APP rats display an increase in the number of microglia recruited towards CA1 hippocampal Amyloid-beta burdened-neurons; AF710B treatment normalized the number of microglial cells to that of untreated wt rats, pointing to anti-inflammatory properties. Conclusions: With M1/sigma-1 activity as well as putative disease-modifying properties at very low doses, AF710B is a promising novel therapeutic agent for the treatment of AD. [1] Fisher et al, 2015 Neurodegener Dis. DOI:10.1159/000440864.