- Email: [email protected]
Involvement of the regulation of X-linked inhibitor of apoptosis protein by GSK3
e26 P4-324
Poster Presentations HUMAN PHOTOSYNTHESIS AND ITS IMPACT IN ALZHEIMER’S AND OTHER NEURODEGENERATIVE DISEASES
Arturo Solis-Herrera, Maria del Carmen Arias-Esparza, Ruth Isabel SolisArias, Paola E. Solis-Arias, Martha Patricia Solis-Arias, Human Photosynthesis Study Center. S.C. Life’s Sciences Division., Aguascalientes, Mexico. Contact e-mail: [email protected] Background: The main source of energy of the eukaryotic cell is the water. Until today, that human tissues have the capability to take hydrogen (that is an energy carrier) from water, as vegetables are, is unknown. However, our studies about the three main causes of blindness, allow us to detect that melanin is the human chlorophyll. Objective: Demonstrate that energy from water is a very important source of hydrogen in the eukaryotic cell. Methods: Systematic review and meta analysis. Conclusion: This amazing compound absorbs photonic energy and transforms it in chemical energy. The reaction in chlorophyll is: 2H2O / 2H2 D O2, and into melanin the reaction is: 2H2O 4 2H2 D O2, and the slight difference between them is the hallmark of the life, because it is not only to dissociate and dissociate, because the hydrogen (and the energy that it carrier) is the really important, and the dangerous is the oxygen, recall its toxic at any level. Furthermore, melanin could increase the oxygen concentration until 97 %, but no more, because at his point, the reaction turn to another side and begin to form water and produce simultaneously a flux of electrons (electricity). In many diseases without an anatomical substrate, like Alzheimer’s and other neurodegenerative process of unknown etiology, it’s very kindly that the ground alteration is an acute or chronic shortage of hydrogen from water. The main consistent finding should be mitochondrial alterations, because hydrogen is the precise compound that drives ATP synthase, and when the hydrogen supply is not adequate, then the main function of ATP synthase (synthesis of ATP) would be damage.
P4-325
ELND005 (SCYLLO-INOSITOL), THE b-AMYLOID ANTI-AGGREGATION THERAPEUTIC, DEMONSTRATES ROBUST BRAIN UPTAKE IN RATS FOLLOWING ORAL ADMINISTRATION
Kevin Quinn1, Beth Brigham1, Ferdie Soriano1, Bruce Connop2, John Michael Sauer1, 1Elan Pharmaceuticals, South San Francisco, CA, USA; 2 Transition Therapeutics, Toronto, ON, Canada Background: ELND005 (scyllo-inositol) is being investigated in the clinic as a potential b-Amyloid anti-aggregation therapy for Alzheimer’s Disease. Preclinical studies (McLaurin, et al., Nature Medicine, July 2006) in TgCRND8 transgenic mice demonstrated both prophylactic and therapeutic properties. Pathological and cognitive abnormalities, and animal survivability were significantly improved with ELND005 treatment. In chronic toxicology studies,
ELND005 was well tolerated in rats and dogs. Pharmacokinetic studies in these species support that ELND005 is completely orally bioavailable. The current study investigated the pharmacokinetics of ELND005 in the CNS. Methods: Male Sprague-Dawley rats (N ¼ 3/dose group/time-point) were orally gavaged with ELND005 at 5, 15 and 30mg/kg twice daily for five days. Plasma, cisternal CSF and frontal cortices were collected at 1, 3, 6, 12 and 24-hours following the last dose. Concentrations were determined by GC/MS. Results: At steadystate, plasma exposure (Cmax and AUC0-24h) to ELND005 increased in a less than proportional manner to dose. Similar proportionality was observed in the CSF and brain. CSF exposure was about half the exposure of plasma, while brain exposure was ten-fold greater than plasma. Absorption was rapid with Tmax ranging 1 to 3hours in all three tissues. Furthermore, CSF concentrations appeared to decay in parallel with plasma, while brain concentrations were constant over the sampling period. Conclusions: ELND005 was shown to readily cross the blood-brain and blood-CSF barriers in rats. Exposure of ELND005 to brain was ten-fold greater than plasma exposure and remained constant. Furthermore, brain concentrations were increased four, eight and twelve-fold over endogenous scyllo-inositol following dosing with ELND005. These observations suggest that the CNS is a deep tissue reservoir. When steady-state systemic exposures in the rat and dog were compared to human, they scaled allometrically and suggest that dispositional mechanisms (i.e., active transport) are conserved across species. This observation is further supported by Garzone et al. (this meeting), where the ratio of CSF to plasma and brain to plasma in humans was similar to those in the rat. P4-326
INVOLVEMENT OF THE REGULATION OF X-LINKED INHIBITOR OF APOPTOSIS PROTEIN BY GSK3
Kiyoko Kato, Student, Toshihisa Tanaka, Golam Sadik, Kentaro Yanagi, Masatoshi Takeda, Osaka University, Suita, Japan Background: Glycogen Synthase Kinase 3 (GSK3) is known as one of kinases involved in the hyper-phosphorylation of tau, memory impairment, and the increased production of Ab. Moreover active GSK3 might be potential for execution of apoptosis by phosphorylation of Bax and MCL-1. Therefore inhibition of GSK3 might be cytoprotective, and further mechanisms should be analyzed because GSK3 is a multi-functional player in signal transduction pathways. X-linked inhibitor of apoptosis (XIAP) has been shown to bind to caspase-3, -7 and -9 resulting in inhibition of caspase activities, and to promote their ubiquitination resulting in the degradation. In the present study, we studied the involvement of GSK3 in the regulation of expression levels of XIAP. Methods: To examine the effects of inhibition of GSK3 on the expression levels of XIAP, SH-SY5Y human neuroblastoma cells were treated with lithium or Kenpaullone, inhibitors to GSK3, and cell lysates were analyzed by western blots. To explore the mechanism of the increased protein level of XIAP, RT-PCR was performed to determine the transcription level. Further the degradation process of XIAP was examined by western blots analysis of SH-SY5Y cells that were pretreated with cycloheximide, a protein synthesis inhibitor, followed by treatment of cells with inhibitors to GSK3. Results: GSK3 inhibition increased the protein level of XIAP in SH-SY5Y cells. Transcription was not changed, however in the presence of cycloheximide protein levels of XIAP was increased suggesting that degradation of XIAP was attenuated by inhibition of GSK3. Conclusions: GSK3 activity might affect protein levels of XIAP presumably by regulating the degradation, but not the transcription. P4-327
FOUNDATION FOR THE NATIONAL INSTITUTES OF HEALTH: FACILITATING THE DEVELOPMENT AND QUALIFICATION OF ALZHEIMER’S DISEASE BIOMARKERS THROUGH NOVEL PUBLICPRIVATE PARTNERSHIPS
David B. Lee, MPA., Foundation for NIH, Bethesda, MD, USA Background: The Foundation for the National Institutese of Health (FNIH) is the sole entity authorized by the U.S. Congress to raise private funds in support of NIH’s mission of improving health through scientific discovery and translational research. Since 1996, FNIH has raised over $410 million towards more
Poster Presentations HUMAN PHOTOSYNTHESIS AND ITS IMPACT IN ALZHEIMER’S AND OTHER NEURODEGENERATIVE DISEASES
Arturo Solis-Herrera, Maria del Carmen Arias-Esparza, Ruth Isabel SolisArias, Paola E. Solis-Arias, Martha Patricia Solis-Arias, Human Photosynthesis Study Center. S.C. Life’s Sciences Division., Aguascalientes, Mexico. Contact e-mail: [email protected] Background: The main source of energy of the eukaryotic cell is the water. Until today, that human tissues have the capability to take hydrogen (that is an energy carrier) from water, as vegetables are, is unknown. However, our studies about the three main causes of blindness, allow us to detect that melanin is the human chlorophyll. Objective: Demonstrate that energy from water is a very important source of hydrogen in the eukaryotic cell. Methods: Systematic review and meta analysis. Conclusion: This amazing compound absorbs photonic energy and transforms it in chemical energy. The reaction in chlorophyll is: 2H2O / 2H2 D O2, and into melanin the reaction is: 2H2O 4 2H2 D O2, and the slight difference between them is the hallmark of the life, because it is not only to dissociate and dissociate, because the hydrogen (and the energy that it carrier) is the really important, and the dangerous is the oxygen, recall its toxic at any level. Furthermore, melanin could increase the oxygen concentration until 97 %, but no more, because at his point, the reaction turn to another side and begin to form water and produce simultaneously a flux of electrons (electricity). In many diseases without an anatomical substrate, like Alzheimer’s and other neurodegenerative process of unknown etiology, it’s very kindly that the ground alteration is an acute or chronic shortage of hydrogen from water. The main consistent finding should be mitochondrial alterations, because hydrogen is the precise compound that drives ATP synthase, and when the hydrogen supply is not adequate, then the main function of ATP synthase (synthesis of ATP) would be damage.
P4-325
ELND005 (SCYLLO-INOSITOL), THE b-AMYLOID ANTI-AGGREGATION THERAPEUTIC, DEMONSTRATES ROBUST BRAIN UPTAKE IN RATS FOLLOWING ORAL ADMINISTRATION
Kevin Quinn1, Beth Brigham1, Ferdie Soriano1, Bruce Connop2, John Michael Sauer1, 1Elan Pharmaceuticals, South San Francisco, CA, USA; 2 Transition Therapeutics, Toronto, ON, Canada Background: ELND005 (scyllo-inositol) is being investigated in the clinic as a potential b-Amyloid anti-aggregation therapy for Alzheimer’s Disease. Preclinical studies (McLaurin, et al., Nature Medicine, July 2006) in TgCRND8 transgenic mice demonstrated both prophylactic and therapeutic properties. Pathological and cognitive abnormalities, and animal survivability were significantly improved with ELND005 treatment. In chronic toxicology studies,
ELND005 was well tolerated in rats and dogs. Pharmacokinetic studies in these species support that ELND005 is completely orally bioavailable. The current study investigated the pharmacokinetics of ELND005 in the CNS. Methods: Male Sprague-Dawley rats (N ¼ 3/dose group/time-point) were orally gavaged with ELND005 at 5, 15 and 30mg/kg twice daily for five days. Plasma, cisternal CSF and frontal cortices were collected at 1, 3, 6, 12 and 24-hours following the last dose. Concentrations were determined by GC/MS. Results: At steadystate, plasma exposure (Cmax and AUC0-24h) to ELND005 increased in a less than proportional manner to dose. Similar proportionality was observed in the CSF and brain. CSF exposure was about half the exposure of plasma, while brain exposure was ten-fold greater than plasma. Absorption was rapid with Tmax ranging 1 to 3hours in all three tissues. Furthermore, CSF concentrations appeared to decay in parallel with plasma, while brain concentrations were constant over the sampling period. Conclusions: ELND005 was shown to readily cross the blood-brain and blood-CSF barriers in rats. Exposure of ELND005 to brain was ten-fold greater than plasma exposure and remained constant. Furthermore, brain concentrations were increased four, eight and twelve-fold over endogenous scyllo-inositol following dosing with ELND005. These observations suggest that the CNS is a deep tissue reservoir. When steady-state systemic exposures in the rat and dog were compared to human, they scaled allometrically and suggest that dispositional mechanisms (i.e., active transport) are conserved across species. This observation is further supported by Garzone et al. (this meeting), where the ratio of CSF to plasma and brain to plasma in humans was similar to those in the rat. P4-326
INVOLVEMENT OF THE REGULATION OF X-LINKED INHIBITOR OF APOPTOSIS PROTEIN BY GSK3
Kiyoko Kato, Student, Toshihisa Tanaka, Golam Sadik, Kentaro Yanagi, Masatoshi Takeda, Osaka University, Suita, Japan Background: Glycogen Synthase Kinase 3 (GSK3) is known as one of kinases involved in the hyper-phosphorylation of tau, memory impairment, and the increased production of Ab. Moreover active GSK3 might be potential for execution of apoptosis by phosphorylation of Bax and MCL-1. Therefore inhibition of GSK3 might be cytoprotective, and further mechanisms should be analyzed because GSK3 is a multi-functional player in signal transduction pathways. X-linked inhibitor of apoptosis (XIAP) has been shown to bind to caspase-3, -7 and -9 resulting in inhibition of caspase activities, and to promote their ubiquitination resulting in the degradation. In the present study, we studied the involvement of GSK3 in the regulation of expression levels of XIAP. Methods: To examine the effects of inhibition of GSK3 on the expression levels of XIAP, SH-SY5Y human neuroblastoma cells were treated with lithium or Kenpaullone, inhibitors to GSK3, and cell lysates were analyzed by western blots. To explore the mechanism of the increased protein level of XIAP, RT-PCR was performed to determine the transcription level. Further the degradation process of XIAP was examined by western blots analysis of SH-SY5Y cells that were pretreated with cycloheximide, a protein synthesis inhibitor, followed by treatment of cells with inhibitors to GSK3. Results: GSK3 inhibition increased the protein level of XIAP in SH-SY5Y cells. Transcription was not changed, however in the presence of cycloheximide protein levels of XIAP was increased suggesting that degradation of XIAP was attenuated by inhibition of GSK3. Conclusions: GSK3 activity might affect protein levels of XIAP presumably by regulating the degradation, but not the transcription. P4-327
FOUNDATION FOR THE NATIONAL INSTITUTES OF HEALTH: FACILITATING THE DEVELOPMENT AND QUALIFICATION OF ALZHEIMER’S DISEASE BIOMARKERS THROUGH NOVEL PUBLICPRIVATE PARTNERSHIPS
David B. Lee, MPA., Foundation for NIH, Bethesda, MD, USA Background: The Foundation for the National Institutese of Health (FNIH) is the sole entity authorized by the U.S. Congress to raise private funds in support of NIH’s mission of improving health through scientific discovery and translational research. Since 1996, FNIH has raised over $410 million towards more