- Email: [email protected]
Amyloidogenesis targeting small molecule as a drug candidate
S532 P3-273
Poster Presentations P3 DISCOVERY OF NOVEL, POTENT BACE INHIBITORS WITH CENTRAL ACTIVITY FOR THE TREATMENT OF ALZHEIMER’S DISEASE
Lynn A. Hyde1, Prescott T. Leach1, Qi Zhang1, Giuseppe Terracina1, Bonnie J. Werner1, Robert A. Hodgson1, Cinzia Cantu1, Xia Chen1, Maxine Chen1, Jeffrey Misiaszek1, Yusheng Wu1, Suresh Babu2, Tao Guo2, Dawit Tadesse2, Jack Scott1, Wei Li1, Lili Zhang1, Eric M. Parker1, Andrew W. Stamford1, Matthew E. Kennedy1, 1Merck Research Laboratories, Kenilworth, NJ, USA; 2Ligand Pharmaceuticals, Inc, Cranbury, NJ, USA. Contact e-mail: [email protected] Background: One of the neuropathological hallmarks of Alzheimer’s disease is the presence of b-amyloid (Ab)-containing neuritic plaques. Ab is produced by the proteolytic cleavage of the amyloid precursor protein (APP) by b- and g-secretase enzymes. Thus, inhibition of b-secretase (BACE1) is a promising disease modifying approach for Alzheimer’s disease. Several high affinity BACE inhibitors have been described that reduce plasma Ab in vivo. Although this may be sufficient to provide clinical benefit (e.g., via peripheral sink), our goal was to identify BACE inhibitors that substantially reduced (>25%) Ab in the brain. Methods: To support ongoing medicinal chemistry SAR efforts to identify BACE inhibitors that would reduce Ab in the brain, we used lowering of Ab1-40 in cerebrospinal fluid (CSF) as a marker of CNS BACE1 inhibition. We also switched our in vivo screening model from transgenic CRND8 mice expressing the Swedish and Indiana mutations in the human APP gene to non-transgenic rats and developed a sensitive rat Ab1-40 immunoassay assay to support the in vivo screen. Results: Our ex vivo binding studies and studies with heterozygous BACE1 mice supported the conclusion that BACE is not the rate limiting step for Ab synthesis in the rodent brain and thus high occupancy of the BACE enzyme is likely required for brain efficacy. After focusing our SAR efforts around robust Ab-lowering in rat CSF, several novel, potent, high affinity, orally available, brain penetrant BACE inhibitors were identified that substantially lowered CSF Ab and more importantly, inhibited Ab in the rat cortex. Based on our experience, brain exposures well in excess of inhibitor cell potency alone did not always result in substantial lowering of CSF or cortical Ab, but that high brain exposure combined with sustained and substantial CSF efficacy was needed to observe Ab-lowering in the cortex. Conclusions: Thus, we have identified several novel, potent BACE inhibitors with excellent Ab-lowering activity in CSF and the brain which may be of use as disease modifying Alzheimer’s disease therapeutics. P3-274
THE PLANT-DERIVED POLYPHENOL TANNIC ACID MITIGATES ALZHEIMER-LIKE PATHOLOGY
was continued for a period of 6 months. Results: Tannic acid administration resulted in significant improvement of transgene-associated behavioral impairment including hyperactivity, impaired object recognition, and defective spatial reference memory, but had no effect on cognitive ability of non-transgenic mice. Furthermore, brain parencymal and cerebral vascular b-amyloid deposits and abundance of Ab peptides were significantly reduced in tannic acid-treated PSAPP mice. These effects were associated with decreased cleavage of the b-C-terminal APP fragment, reduction of b-site APP cleaving enzyme 1 protein expression and activity, attenuated b-amyloid plaque-associated gliosis, and decreased mRNA expression of proinflammatory cytokines such as IL-1b and TNF-a. Conclusions: These results raise the possibility that tannic acid dietary supplementation may be a promising prophylaxis for AD. P3-275
OPTIMIZATION OF THE TREATMENT REGIMEN WITH ACTIVE Aß IMMUNOTHERAPY CAD106 IN ALZHEIMER PATIENTS
Ana Graf1, Niels Andreasen2, M. E. Riviere1, Jacqueline Ros1, Jessie Moreau1, Jeffrey Sevigny1, Peter Quarg1, Angelika Caputo1, Luca Finelli1, Bengt Winblad2, Georges Imbert1, 1Novartis Pharma AG, Basel, Switzerland; 2Karolinska Universitetssjukhuset, Huddinge, Sweden. Contact e-mail: [email protected] Background: CAD106 is an active immunotherapy being developed for Alzheimers Disease (AD), which comprises the amyloid (Aß) 1-6 peptide coupled to the Qß virus-like particle. In animals, CAD106 induced Aßantibody titers without activating Aß-reactive T-cells. Administration of CAD106 to APP transgenic mice showed a reduction of amyloid accumulation in the brain. Methods: Two regimens were tested in two Phase IIa studies, with the objective to assess safety, tolerability and immunogenicity. These studies were 52-week, multi-center, randomized, double-blind, placebo-controlled, parallel group studies in patients with mild AD. CAD106 dose of 150 mg was administered subcutaneously (s.c.) at weeks 0/6/12 in Study 1, and s.c. or intramuscularly (i.m.) at weeks 0/2/6 in Study 2. Results: At baseline, out of 58 patients, none had Aß-specific IgG antibody titers above the Lower Limit of Quantitation (LLOQ, 8.9 units) of the assay. Aß IgG antibody response was defined as increase above 16 units (LLOQ + 3 SD) at least once during the study. Following treatment with CAD106 in Study 1, antibody response was observed in 20/22 patients. Similar mean peak titers were achieved following both the second and third injection (week 8: 44 units, week 14: 40 units). In addition, antibodies were detected for longer than 6 months. Preliminary data from Study 2 are available. These data show a single antibody titer peak at week 8 of similar magnitude at the same time-point in Study 1. Responder rate and duration of response seem also to be comparable at this early stage. Antibody titer profiles with s.c. and i.m. routes of administration from Study 2 will be presented. Conclusions: Preliminary data from these studies confirm previous information from Phase I that injection at week 2 does not enhance antibody response. A week 0/6/12 regimen is therefore selected for the next study. Further injections at 12-week intervals are also being evaluated, along with addition of adjuvants.
Takashi Mori1, Naoki Koyama1, Kavon Rezai-Zadeh2, Nobuto Kakuda3, Haruyasu Yamaguchi4, Gary W. Arendash5,6, Jun Tan7, Terrence Town2,8, 1 Saitama Medical Center/University, Kawagoe, Japan; 2Departments of Biomedical Sciences and Neurosurgery, Maxine Dunitz Neurosurgical Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; 3 Immuno-Biological Laboratories Co., Ltd., Fujioka, Japan; 4Gunma University School of Health Sciences, Maebashi, Japan; 5Florida Alzheimer’s Disease Research Center, University of South Florida, Tampa, FL, USA; 6 Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, FL, USA; 7Department of Psychiatry and Behavioral Medicine, University of South Florida, Tampa, FL, USA; 8 Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA, USA. Contact e-mail: t_mori@ saitama-med.ac.jp
YoungSoo Kim1, Seong Rim Byeon1, HyeYun Kim1, Ji Hoon Lee1, Hana Hwang1, Jeiwon Cho1, Young-Gil Ahn2, Young Hoon Kim2, Maeng Sup Kim2, Gwan Sun Lee2, Inhee Mook-Jung3, Dong Jin Kim1, 1 Korea Institute of Science & Technology, Seoul, Republic of Korea; 2Hanmi Pharm. Co., Ltd., Seoul, Republic of Korea; 3Seoul National University, Seoul, Republic of Korea. Contact e-mail: [email protected]
Background: Amyloid precursor protein (APP) proteolysis is essential for production of amyloid-b (Ab) peptides that form b-amyloid deposits in brains of Alzheimer’s disease (AD) patients. Owing to their anti-amyloidogenic properties, recent focus has been given to a group of naturally occurring polyphenols known as flavonoids. Methods: We orally administered the polymeric polyphenol tannic acid to the transgenic PSAPP (APPsw, PSEN1dE9) mouse model of AD and evaluated impact on cognitive impairment and AD-like pathology. Treatment commenced at 6 months of age and
Background: Since cholinergic deficit in the central nervous system has been observed in AD patients, inhibitors for acetylcholine esterase (AchE) such as tacrin, donepezil, and rivastigmine are used for the clinical treatment of AD symptoms in current therapeutic approaches. Although these are the only FDA-approved AD therapies on the market, they cannot solve fundamental problems but can be only palliative solutions. Pathomorphologically important hallmarks in AD are senile plaques (SP) composed of amyloid b (Ab) and neurofibrillary tangles (NFTs). Therefore, reducing deposition of Ab plaques in
P3-276
AMYLOIDOGENESIS TARGETING SMALL MOLECULE AS A DRUG CANDIDATE
Poster Presentations P3 the brain is believed to be pivotal to treat the AD patients. Based on Ab hypothesis, Ab(1-42) monomers released from APP by b- and g-secretases are gradually aggregated to oligomers, protofibrils, and fibrils. Ab(1-42) aggregates such as oligomers and fibrils deposit in the brain causing strong neuronal toxicity. In addition, it was recently reported that oligomers might be more toxic than fibrils, and the oligomers are becoming the target for the development of AD treatments. Methods: Here we report pharmacology, ADME and toxicity tests of a novel aminostyrylbenzofuran derivative with potent inhibitory activities for amyloidogenesis. Results: KMS88009 showed outstanding results in in vitro Inhibition activities of formation of Ab42 plague, fear conditioning, water maze, Y-maze, noble object recognition, hERG assay, ADME and toxicity. Conclusions: In particular, KMS88009 is a promising AD drug candidate as an amyloidogenesis modulator. P3-277
IVIG TREATMENT IN ALZHEIMER’S AND LEWY BODY DISEASE: POTENTIAL DISEASE DELAYING EFFECT
Junko Hara1,2, Kaitlin Masatsugu2, William R. Shankle1,2, 1Medical Care Corporation, Irvine, CA, USA; 2Shankle Clinic, Irvine, CA, USA. Contact e-mail: [email protected] Background: Intravenous immunoglobulin (IVIg) therapy has shown treatment efficacy in patients with Alzheimer’s disease (AD) in a 6-month trial of 5 patients, in an 18-month, open-label trial of 8 patients, and in a retrospective study of 10 patients. In two retrospective analyses_one in the USA and one in Sweden_patients treated with IVIG for autoimmune disorders had reduced risk of developing dementia or AD. In the present study, we evaluated treatment efficacy of IVIG for a series of AD and Lewy body disease (LBD) patients treated for up to 55 months. Methods: Patient with AD (n ¼ 13) and with LBD (n ¼ 4) received IVIg for an average of 26.4 (std. ¼ 16.8) and 15.6 (std. ¼ 17.2) months. All patients were treated with cholinesterase inhibitors and/or memantine before and during IVIG. The primary functional and cognitive measures were the Functional Assessment Staging Test (FAST) and the MCI Screen (MCIS). Other measures included ADAS-Cog, MMSE, NPI, ADL, CGIC, ApoE genotype, plus baseline and endpoint CSF Tau and Ab42 levels when data were available. Results: Compared to the expected, untreated duration for each FAST stage, IVIg treatment delayed FAST stage progression by an average of 30.7 (std. ¼ 23.8) and 6.6 (std. ¼ 18.2) months for AD and LBD patients respectively. This disease-delaying effect was greater when IVIG was started in earlier AD stages (FAST ¼ 4,5), and was not related to ApoE genotype. Cognitively, the MCIS showed that IVIG slowed rate of decline, especially for AD patients. Conclusions: This study suggests that IVIG has a disease-delaying effect for AD and LBD, particularly when started in milder stages. A mechanistic hypothesis for delaying AD and LBD progression is the binding of IgG to abnormally folded proteins of oligomeric beta amyloid in AD, and alpha synuclein in LBD. More sophisticated analytical methods are being used to evaluate potential confounding influences on these results.
conditioned, unlabeled media and 50% unconditioned 13C6 labeled media. As cells produce Abeta the stable isotope is incorporated into newly synthesized Abeta. Results: By measuring the ratio of labeled to unlabeled Abeta for each of the C-terminally differentiated Abeta isoforms at 4, 8, 12 and 24 hours after the media change we can estimate the production curves for each of these isoforms. Subsequently, we tested the effects that a variety of publicly available gamma-secretase modulating drugs have on the production curves of the Abeta isoforms. Conclusions: Our data show that the SILK-Abeta isoform methodology allows for precise quantification of the rate of production of different Abeta isoforms in cultured cells. This method has application for measuring the in vivo metabolism of Abeta isoforms in humans. P3-279
EFFECT OF GAMMA-SECRETASE MODULATING DRUGS ON ABETA ISOFORM PRODUCTION RATES FROM CULTURED CELLS USING STABLE ISOTOPE LABELING KINETICS AND MASS SPECTROMETRY
EFFECT OF TIBET-MEDICINE ON CHANGES OF SERUM b-AMYLOID PROTEIN AND CYTOKINE LEVELS IN PATIENTS WITH ALZHEIMER’S DISEASE
Aiqin Zhu, Yide Chu, Guofeng Li, Qinghai Provincial Hospital,Institution of Geriatrics, Xining, China. Contact e-mail: [email protected] Background: To study of Ratanasampil (RNSP)which is traditional Chinese Tibet-medicines on changes of serum b-amyloid protein and cytokine interleukin levels and tumor necrosis factor alpha(TNF-a)in patients with mild to moderate Alzheimer’s disease (AD) at high altitude. Methods: One hundred cases of patients with AD at high altitude of 2,600m above sea levels were divided into two groups randomly including treated by Ratanasampil and Piracetam, respectively. The whole course of treatment lasted for 12 weeks. Mini Mental State Examination (MMSE), Alzheimer’s disease Assessment Scale-cognitive subscale (ADAS-cog) and Activity of Daily Living Scale (ADL) were taken to evaluated the efficacy. Serum levels of amyloid peptides (Ab40 and Ab40) were measured by ELISA assay. The radioimmunologic assay was used to determine the levels of IL-1b,IL-2,IL-6,IL-8 and TNF-a from serum. ECG and laboratorial tests were measured every 4 weeks for safety. Results: The scores of MMSE, ADAS-cog and ADL in treated group were significantly improved at 12 wk after Ratanasampil treatment (p value respectively < 0.01, 0.01, 0.05), while no significant change in the Piracetam treated group(p > 0.05). The levels of TNF-a, IL-1b, IL-6 and Ab42 after Ratanasampil treatment were significantly lower than those in Piracetam group (p <0. 01). There was a decrease trend of the Ab42/Ab40 ratio after Ratanasampil-treated patients (p < 0. 05, p <0.01). The serum Ab42 had a strong correlation with TNF-a, IL-1b and IL-6. We could not find significant difference of Ab40 and IL-8 between Ratanasampil and Piracetam group. No obvious drug side effect happened on the groups. Conclusions: The serum TNF-a, IL-1b and IL-6 reduction after Ratanasampil treatment may lead to decrease Ab42 production in AD patients. Ratanasampil may decrease the Ab42/Ab40 ratio and slowed down the progress of AD. It is effective in treating patients with mild to moderate AD at high altitude and may improve the learning and memory ability in those. It was well tolerated and safe. P3-280
P3-278
S533
CHARACTERIZATION OF TAT-CONJUGATED B6C15 PEPTIDE WHICH IS A COMMON MIMOTOPE OF Aß42-FIBRIL-SPECIFIC HUMAN SCFVS IN A FLOW OF Aß42 FIBRILLATION
Tim West, Mary S. Holubasch, Andrew C. Paoletti, C2N Diagnostics, Saint Louis, MO, USA. Contact e-mail: [email protected]
Masaaki Nishimura, Yuka Dochi, Koichi Tanaka, Makoto Yamaguchi, Risa Abe, Yuya Yamaguchi, Shuhei Hashiguchi, Yuji Ito, Kazuhisa Sugimura, Kagoshima University, Kagoshima, Japan. Contact e-mail: [email protected]
Background: Several drugs have been proposed to modulate the cleavage site of gamma-secretase within amyloid precursor protein giving rise to production of different ratios and amounts of amyloid-beta (Abeta) isoforms with various C-termini. However, studies to date have focused on static measurements of Abeta isoform concentrations in biological fluids such as cell culture media or brain homogenates. Methods: We have developed a stable isotope labeling kinetic (SILK-Abeta isoform) methodology for measuring production of the C-terminally differentiated Abeta isoforms and have applied this methodology to the measurement of production rates of these isoforms from neuroblastoma cells over-expressing APP. Cells are grown in unlabeled media for 24 hours after which they are switched to media that contains 50%
Background: Aggregation and fibril formation of amyloid-ß42 (Aß42) peptides is crucial events in the phathogenesis of Alzheimer’s disease. Recently, we have established conformation dependent human antibody, B6, which binds to Aß42 fibril, but not to soluble form of Aß42, inhibiting Aß42 fibril formation. We have identified a mimotope of B6, B6-C15, using the random peptide displaying phage library (PhD.C7C). We synthesized B6-C15 peptide conjugated with biotinylated TAT at the N-terminus (TAT-B6-C15). It was demonstrated that this peptide inhibited Aß42 fibrillation by binding to prefibrillar oligomer, but not to freshly prepared monomer Aß42 nor its fibril form. In this study, we investigated the molecular mechanism of Aß42 fibril formation by using of TAT-B6-C15 peptide. Methods: Aß42
Poster Presentations P3 DISCOVERY OF NOVEL, POTENT BACE INHIBITORS WITH CENTRAL ACTIVITY FOR THE TREATMENT OF ALZHEIMER’S DISEASE
Lynn A. Hyde1, Prescott T. Leach1, Qi Zhang1, Giuseppe Terracina1, Bonnie J. Werner1, Robert A. Hodgson1, Cinzia Cantu1, Xia Chen1, Maxine Chen1, Jeffrey Misiaszek1, Yusheng Wu1, Suresh Babu2, Tao Guo2, Dawit Tadesse2, Jack Scott1, Wei Li1, Lili Zhang1, Eric M. Parker1, Andrew W. Stamford1, Matthew E. Kennedy1, 1Merck Research Laboratories, Kenilworth, NJ, USA; 2Ligand Pharmaceuticals, Inc, Cranbury, NJ, USA. Contact e-mail: [email protected] Background: One of the neuropathological hallmarks of Alzheimer’s disease is the presence of b-amyloid (Ab)-containing neuritic plaques. Ab is produced by the proteolytic cleavage of the amyloid precursor protein (APP) by b- and g-secretase enzymes. Thus, inhibition of b-secretase (BACE1) is a promising disease modifying approach for Alzheimer’s disease. Several high affinity BACE inhibitors have been described that reduce plasma Ab in vivo. Although this may be sufficient to provide clinical benefit (e.g., via peripheral sink), our goal was to identify BACE inhibitors that substantially reduced (>25%) Ab in the brain. Methods: To support ongoing medicinal chemistry SAR efforts to identify BACE inhibitors that would reduce Ab in the brain, we used lowering of Ab1-40 in cerebrospinal fluid (CSF) as a marker of CNS BACE1 inhibition. We also switched our in vivo screening model from transgenic CRND8 mice expressing the Swedish and Indiana mutations in the human APP gene to non-transgenic rats and developed a sensitive rat Ab1-40 immunoassay assay to support the in vivo screen. Results: Our ex vivo binding studies and studies with heterozygous BACE1 mice supported the conclusion that BACE is not the rate limiting step for Ab synthesis in the rodent brain and thus high occupancy of the BACE enzyme is likely required for brain efficacy. After focusing our SAR efforts around robust Ab-lowering in rat CSF, several novel, potent, high affinity, orally available, brain penetrant BACE inhibitors were identified that substantially lowered CSF Ab and more importantly, inhibited Ab in the rat cortex. Based on our experience, brain exposures well in excess of inhibitor cell potency alone did not always result in substantial lowering of CSF or cortical Ab, but that high brain exposure combined with sustained and substantial CSF efficacy was needed to observe Ab-lowering in the cortex. Conclusions: Thus, we have identified several novel, potent BACE inhibitors with excellent Ab-lowering activity in CSF and the brain which may be of use as disease modifying Alzheimer’s disease therapeutics. P3-274
THE PLANT-DERIVED POLYPHENOL TANNIC ACID MITIGATES ALZHEIMER-LIKE PATHOLOGY
was continued for a period of 6 months. Results: Tannic acid administration resulted in significant improvement of transgene-associated behavioral impairment including hyperactivity, impaired object recognition, and defective spatial reference memory, but had no effect on cognitive ability of non-transgenic mice. Furthermore, brain parencymal and cerebral vascular b-amyloid deposits and abundance of Ab peptides were significantly reduced in tannic acid-treated PSAPP mice. These effects were associated with decreased cleavage of the b-C-terminal APP fragment, reduction of b-site APP cleaving enzyme 1 protein expression and activity, attenuated b-amyloid plaque-associated gliosis, and decreased mRNA expression of proinflammatory cytokines such as IL-1b and TNF-a. Conclusions: These results raise the possibility that tannic acid dietary supplementation may be a promising prophylaxis for AD. P3-275
OPTIMIZATION OF THE TREATMENT REGIMEN WITH ACTIVE Aß IMMUNOTHERAPY CAD106 IN ALZHEIMER PATIENTS
Ana Graf1, Niels Andreasen2, M. E. Riviere1, Jacqueline Ros1, Jessie Moreau1, Jeffrey Sevigny1, Peter Quarg1, Angelika Caputo1, Luca Finelli1, Bengt Winblad2, Georges Imbert1, 1Novartis Pharma AG, Basel, Switzerland; 2Karolinska Universitetssjukhuset, Huddinge, Sweden. Contact e-mail: [email protected] Background: CAD106 is an active immunotherapy being developed for Alzheimers Disease (AD), which comprises the amyloid (Aß) 1-6 peptide coupled to the Qß virus-like particle. In animals, CAD106 induced Aßantibody titers without activating Aß-reactive T-cells. Administration of CAD106 to APP transgenic mice showed a reduction of amyloid accumulation in the brain. Methods: Two regimens were tested in two Phase IIa studies, with the objective to assess safety, tolerability and immunogenicity. These studies were 52-week, multi-center, randomized, double-blind, placebo-controlled, parallel group studies in patients with mild AD. CAD106 dose of 150 mg was administered subcutaneously (s.c.) at weeks 0/6/12 in Study 1, and s.c. or intramuscularly (i.m.) at weeks 0/2/6 in Study 2. Results: At baseline, out of 58 patients, none had Aß-specific IgG antibody titers above the Lower Limit of Quantitation (LLOQ, 8.9 units) of the assay. Aß IgG antibody response was defined as increase above 16 units (LLOQ + 3 SD) at least once during the study. Following treatment with CAD106 in Study 1, antibody response was observed in 20/22 patients. Similar mean peak titers were achieved following both the second and third injection (week 8: 44 units, week 14: 40 units). In addition, antibodies were detected for longer than 6 months. Preliminary data from Study 2 are available. These data show a single antibody titer peak at week 8 of similar magnitude at the same time-point in Study 1. Responder rate and duration of response seem also to be comparable at this early stage. Antibody titer profiles with s.c. and i.m. routes of administration from Study 2 will be presented. Conclusions: Preliminary data from these studies confirm previous information from Phase I that injection at week 2 does not enhance antibody response. A week 0/6/12 regimen is therefore selected for the next study. Further injections at 12-week intervals are also being evaluated, along with addition of adjuvants.
Takashi Mori1, Naoki Koyama1, Kavon Rezai-Zadeh2, Nobuto Kakuda3, Haruyasu Yamaguchi4, Gary W. Arendash5,6, Jun Tan7, Terrence Town2,8, 1 Saitama Medical Center/University, Kawagoe, Japan; 2Departments of Biomedical Sciences and Neurosurgery, Maxine Dunitz Neurosurgical Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; 3 Immuno-Biological Laboratories Co., Ltd., Fujioka, Japan; 4Gunma University School of Health Sciences, Maebashi, Japan; 5Florida Alzheimer’s Disease Research Center, University of South Florida, Tampa, FL, USA; 6 Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, FL, USA; 7Department of Psychiatry and Behavioral Medicine, University of South Florida, Tampa, FL, USA; 8 Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA, USA. Contact e-mail: t_mori@ saitama-med.ac.jp
YoungSoo Kim1, Seong Rim Byeon1, HyeYun Kim1, Ji Hoon Lee1, Hana Hwang1, Jeiwon Cho1, Young-Gil Ahn2, Young Hoon Kim2, Maeng Sup Kim2, Gwan Sun Lee2, Inhee Mook-Jung3, Dong Jin Kim1, 1 Korea Institute of Science & Technology, Seoul, Republic of Korea; 2Hanmi Pharm. Co., Ltd., Seoul, Republic of Korea; 3Seoul National University, Seoul, Republic of Korea. Contact e-mail: [email protected]
Background: Amyloid precursor protein (APP) proteolysis is essential for production of amyloid-b (Ab) peptides that form b-amyloid deposits in brains of Alzheimer’s disease (AD) patients. Owing to their anti-amyloidogenic properties, recent focus has been given to a group of naturally occurring polyphenols known as flavonoids. Methods: We orally administered the polymeric polyphenol tannic acid to the transgenic PSAPP (APPsw, PSEN1dE9) mouse model of AD and evaluated impact on cognitive impairment and AD-like pathology. Treatment commenced at 6 months of age and
Background: Since cholinergic deficit in the central nervous system has been observed in AD patients, inhibitors for acetylcholine esterase (AchE) such as tacrin, donepezil, and rivastigmine are used for the clinical treatment of AD symptoms in current therapeutic approaches. Although these are the only FDA-approved AD therapies on the market, they cannot solve fundamental problems but can be only palliative solutions. Pathomorphologically important hallmarks in AD are senile plaques (SP) composed of amyloid b (Ab) and neurofibrillary tangles (NFTs). Therefore, reducing deposition of Ab plaques in
P3-276
AMYLOIDOGENESIS TARGETING SMALL MOLECULE AS A DRUG CANDIDATE
Poster Presentations P3 the brain is believed to be pivotal to treat the AD patients. Based on Ab hypothesis, Ab(1-42) monomers released from APP by b- and g-secretases are gradually aggregated to oligomers, protofibrils, and fibrils. Ab(1-42) aggregates such as oligomers and fibrils deposit in the brain causing strong neuronal toxicity. In addition, it was recently reported that oligomers might be more toxic than fibrils, and the oligomers are becoming the target for the development of AD treatments. Methods: Here we report pharmacology, ADME and toxicity tests of a novel aminostyrylbenzofuran derivative with potent inhibitory activities for amyloidogenesis. Results: KMS88009 showed outstanding results in in vitro Inhibition activities of formation of Ab42 plague, fear conditioning, water maze, Y-maze, noble object recognition, hERG assay, ADME and toxicity. Conclusions: In particular, KMS88009 is a promising AD drug candidate as an amyloidogenesis modulator. P3-277
IVIG TREATMENT IN ALZHEIMER’S AND LEWY BODY DISEASE: POTENTIAL DISEASE DELAYING EFFECT
Junko Hara1,2, Kaitlin Masatsugu2, William R. Shankle1,2, 1Medical Care Corporation, Irvine, CA, USA; 2Shankle Clinic, Irvine, CA, USA. Contact e-mail: [email protected] Background: Intravenous immunoglobulin (IVIg) therapy has shown treatment efficacy in patients with Alzheimer’s disease (AD) in a 6-month trial of 5 patients, in an 18-month, open-label trial of 8 patients, and in a retrospective study of 10 patients. In two retrospective analyses_one in the USA and one in Sweden_patients treated with IVIG for autoimmune disorders had reduced risk of developing dementia or AD. In the present study, we evaluated treatment efficacy of IVIG for a series of AD and Lewy body disease (LBD) patients treated for up to 55 months. Methods: Patient with AD (n ¼ 13) and with LBD (n ¼ 4) received IVIg for an average of 26.4 (std. ¼ 16.8) and 15.6 (std. ¼ 17.2) months. All patients were treated with cholinesterase inhibitors and/or memantine before and during IVIG. The primary functional and cognitive measures were the Functional Assessment Staging Test (FAST) and the MCI Screen (MCIS). Other measures included ADAS-Cog, MMSE, NPI, ADL, CGIC, ApoE genotype, plus baseline and endpoint CSF Tau and Ab42 levels when data were available. Results: Compared to the expected, untreated duration for each FAST stage, IVIg treatment delayed FAST stage progression by an average of 30.7 (std. ¼ 23.8) and 6.6 (std. ¼ 18.2) months for AD and LBD patients respectively. This disease-delaying effect was greater when IVIG was started in earlier AD stages (FAST ¼ 4,5), and was not related to ApoE genotype. Cognitively, the MCIS showed that IVIG slowed rate of decline, especially for AD patients. Conclusions: This study suggests that IVIG has a disease-delaying effect for AD and LBD, particularly when started in milder stages. A mechanistic hypothesis for delaying AD and LBD progression is the binding of IgG to abnormally folded proteins of oligomeric beta amyloid in AD, and alpha synuclein in LBD. More sophisticated analytical methods are being used to evaluate potential confounding influences on these results.
conditioned, unlabeled media and 50% unconditioned 13C6 labeled media. As cells produce Abeta the stable isotope is incorporated into newly synthesized Abeta. Results: By measuring the ratio of labeled to unlabeled Abeta for each of the C-terminally differentiated Abeta isoforms at 4, 8, 12 and 24 hours after the media change we can estimate the production curves for each of these isoforms. Subsequently, we tested the effects that a variety of publicly available gamma-secretase modulating drugs have on the production curves of the Abeta isoforms. Conclusions: Our data show that the SILK-Abeta isoform methodology allows for precise quantification of the rate of production of different Abeta isoforms in cultured cells. This method has application for measuring the in vivo metabolism of Abeta isoforms in humans. P3-279
EFFECT OF GAMMA-SECRETASE MODULATING DRUGS ON ABETA ISOFORM PRODUCTION RATES FROM CULTURED CELLS USING STABLE ISOTOPE LABELING KINETICS AND MASS SPECTROMETRY
EFFECT OF TIBET-MEDICINE ON CHANGES OF SERUM b-AMYLOID PROTEIN AND CYTOKINE LEVELS IN PATIENTS WITH ALZHEIMER’S DISEASE
Aiqin Zhu, Yide Chu, Guofeng Li, Qinghai Provincial Hospital,Institution of Geriatrics, Xining, China. Contact e-mail: [email protected] Background: To study of Ratanasampil (RNSP)which is traditional Chinese Tibet-medicines on changes of serum b-amyloid protein and cytokine interleukin levels and tumor necrosis factor alpha(TNF-a)in patients with mild to moderate Alzheimer’s disease (AD) at high altitude. Methods: One hundred cases of patients with AD at high altitude of 2,600m above sea levels were divided into two groups randomly including treated by Ratanasampil and Piracetam, respectively. The whole course of treatment lasted for 12 weeks. Mini Mental State Examination (MMSE), Alzheimer’s disease Assessment Scale-cognitive subscale (ADAS-cog) and Activity of Daily Living Scale (ADL) were taken to evaluated the efficacy. Serum levels of amyloid peptides (Ab40 and Ab40) were measured by ELISA assay. The radioimmunologic assay was used to determine the levels of IL-1b,IL-2,IL-6,IL-8 and TNF-a from serum. ECG and laboratorial tests were measured every 4 weeks for safety. Results: The scores of MMSE, ADAS-cog and ADL in treated group were significantly improved at 12 wk after Ratanasampil treatment (p value respectively < 0.01, 0.01, 0.05), while no significant change in the Piracetam treated group(p > 0.05). The levels of TNF-a, IL-1b, IL-6 and Ab42 after Ratanasampil treatment were significantly lower than those in Piracetam group (p <0. 01). There was a decrease trend of the Ab42/Ab40 ratio after Ratanasampil-treated patients (p < 0. 05, p <0.01). The serum Ab42 had a strong correlation with TNF-a, IL-1b and IL-6. We could not find significant difference of Ab40 and IL-8 between Ratanasampil and Piracetam group. No obvious drug side effect happened on the groups. Conclusions: The serum TNF-a, IL-1b and IL-6 reduction after Ratanasampil treatment may lead to decrease Ab42 production in AD patients. Ratanasampil may decrease the Ab42/Ab40 ratio and slowed down the progress of AD. It is effective in treating patients with mild to moderate AD at high altitude and may improve the learning and memory ability in those. It was well tolerated and safe. P3-280
P3-278
S533
CHARACTERIZATION OF TAT-CONJUGATED B6C15 PEPTIDE WHICH IS A COMMON MIMOTOPE OF Aß42-FIBRIL-SPECIFIC HUMAN SCFVS IN A FLOW OF Aß42 FIBRILLATION
Tim West, Mary S. Holubasch, Andrew C. Paoletti, C2N Diagnostics, Saint Louis, MO, USA. Contact e-mail: [email protected]
Masaaki Nishimura, Yuka Dochi, Koichi Tanaka, Makoto Yamaguchi, Risa Abe, Yuya Yamaguchi, Shuhei Hashiguchi, Yuji Ito, Kazuhisa Sugimura, Kagoshima University, Kagoshima, Japan. Contact e-mail: [email protected]
Background: Several drugs have been proposed to modulate the cleavage site of gamma-secretase within amyloid precursor protein giving rise to production of different ratios and amounts of amyloid-beta (Abeta) isoforms with various C-termini. However, studies to date have focused on static measurements of Abeta isoform concentrations in biological fluids such as cell culture media or brain homogenates. Methods: We have developed a stable isotope labeling kinetic (SILK-Abeta isoform) methodology for measuring production of the C-terminally differentiated Abeta isoforms and have applied this methodology to the measurement of production rates of these isoforms from neuroblastoma cells over-expressing APP. Cells are grown in unlabeled media for 24 hours after which they are switched to media that contains 50%
Background: Aggregation and fibril formation of amyloid-ß42 (Aß42) peptides is crucial events in the phathogenesis of Alzheimer’s disease. Recently, we have established conformation dependent human antibody, B6, which binds to Aß42 fibril, but not to soluble form of Aß42, inhibiting Aß42 fibril formation. We have identified a mimotope of B6, B6-C15, using the random peptide displaying phage library (PhD.C7C). We synthesized B6-C15 peptide conjugated with biotinylated TAT at the N-terminus (TAT-B6-C15). It was demonstrated that this peptide inhibited Aß42 fibrillation by binding to prefibrillar oligomer, but not to freshly prepared monomer Aß42 nor its fibril form. In this study, we investigated the molecular mechanism of Aß42 fibril formation by using of TAT-B6-C15 peptide. Methods: Aß42