CENTRO INTERNACIONAL DE BIOMEDICINA

NEUROCIENCIA

Y MEDICINA TRADUCCIONAL

Dr. R.B. Maccioni Professor of Neurology, F. Medicine, U Chile & Director INTERNATIONAL CENTER FOR BIOMEDICINE (ICC)

Ruta de Presentacin

Alcanzar una saludable longevidad: desafo de la medicina siglo XXI Innovacin: logros del ICC en la comprensin y tratamiento del Alzheimer El aporte y el impacto de las Neurociencias Bsqueda de soluciones mdicas Biomarcadores: solucin hacia la deteccin temprana y diagnstico diferencial. El tratamiento: la tecnologa Brain Up y la estimulacin cognitiva Ensayos clnicos

LOGROS RECIENTES DE ICC HACIA UNA SALUDABLE LONGEVIDAD

ICC se fund en 1989 y hoy est considerado como ...a world class center of excellence. ICC ha sido reconocido por el Programa Human Frontiers entre los 3 centros lderes en el mundo en desarrollo. Descubrimientos sobre el rol del estrs oxidativo en el envejecimiento y
en los trastornos neurodegenerativos. Mecanismos de transduccin de seales en la patogenia del Alzheimer (AD). La hipotesis revitalizada de la protena tau La teora de la neuroinmunomodulacin en el orgen de la enfermedad de Alzheimer Ingeniera de protenas y modelamiento molecular del complejo cdk5/p35. Avances en la comprensin gentica y la farmacologa de la AD. Desarrollo innovador de biomarcadores no invasivos para la AD Nutraceuticos en el tratamiento de la AD 564 publicaciones cientficas, mayoria ISI (7 papers destacados en portadas de revistas de alto impacto). Algunos papers reconocidos entre los ms citados en el tema a nivel mundial Cientificos de ICC obtienen importantes Premios a nivel Mundial por

Compromiso ICC
Triple hlice de la innovacin

Universidad

Industria

Gobierno

Investigaciones en ICC

Enfermedad de Alzheimer

Diagnstico
Frmacos Anti Alzheimer

Biomarcadores

Neuroimagenes

Neuropsicologa

PATENTES
- Molecular markers in the CSF for early diagnosis of Alzheimers disease. Patente U.S.A. (2007) Author: Dr. Ricardo B. Maccioni. Beneficiary: ICC. The solution proposed in this patent document is development of a Diagnosis procedure from CSF samples to determine al algorithm of hyperphosphorylated tau and Ab(1-42) for early diagnosis of Azheimers disease and MCI. Under use throughout the world to detect Alzheimer. - An innovative blood platelets biomarker for early diagnosis of Alzheimers disease Patent PCT/U.S.A.10/55623 and in the U.S.A. Authors: Drs. R.B. Maccioni and G.Faras. Beneficiary: Neuroinnovation. Patent being handled by by Gottielb, Rackmann and Reisman, Attorneys at Law, New York. Status: Published. Product: Diagnosis kit for early detection of AD. - Benzimidazoles as radiotracers for neuroimaging in the diagnosis of neurodegenerative disorders. Authors: Drs. Ricardo B. Maccioni and Leonel Rojo. Beneficiary: Neuroinnovation. PCT Number: PCT/IB2009/006405 and in the U.S.A. EAPI Project CORFO. Status: Published. Potencial Product: PET technology for neuroimaging of Alzheimerss brains - A family of quinolines and their potential usefullness for the treatement of Alzheimers disease. Beneficiary: ICC & Universidad de Chile. Number: Patent 8.198.300 EAPI CORFO Project. Status: APROVED with TITLE CERTIFICATE. Potental Technological Prototype: A compound as a tool to contribute to treatmente of patients with mild Alzheimers Disease. Under technology transfer process. - A novel nutraceutic formulae with neuroprotection and cognitive activities. Authors: Drs. R.B. Maccioni, L.Quiones V. Sandoval, R Sandoval and and I. Saavedra. Beneficiary: ICC- BrainUp Chile. Presented PCT/CL2010/000043 Oct. 2010 at INAPI. Patent in Chile Reg. 01956 /2009. Status: Published in Chile and at the Internacional Office of Patents and in USA, presented Australia, UK, Brazil and Canada . Technological prototype under technology transfer process. - Registration of Intellectual Property and the TradeMark (in Chile) for the software ACTIVAMENTE for psicostimulation and cognitive activity. Corresponds to a software de 6,000 exercises and 50 complex tasks for cognitive intervention. Two versions: 1) Therapeutic version for patents with Alzhemers disease to be used with the therapist. 2) Autonomous subject. Authors: Ps. Alejandra Sekler and Dr. R.B. Maccioni. Beneficiaries: Neuroinnovation Ltd. and Adexus S.A. Product: Software ACTIVAMENTE being

Patentes de Chile en el sistema PCT.

Productividad cientfica y patentes (1999 2009)* Chile Irlanda N. Zelandia Finlandia Pub/ao/millon hab. 117 670 1.097 2.300 Patente/ ao/millon hab. 0.8 67.0 195.0 253.0

Fuente: Base de datos de Conicyt www.conicyt.cl

Porqu el Alzheimer es un problema de salud pblica?

Su prevalencia aumenta drsticamente
2/3 de los casos de Alzheimer y otras demencias viven en el mundo en desarrollo.

Ferri et al., 2005 Kalaria et al., 2008

INTERNATIONAL CENTER FOR BIOMEDICINE

PROBLEMA A RESOLVER
Segn OMS 35.6 millones en el mundo sufren de la enfermedad de Alzheimer (EA) con un impacto en la economa de US$ 600 billones(1) . Para 2030 la poblacin con EA ser 65.6 millones(1). Cada ao 970.000 personas entre 30 - 50 aos desarrollan EA prematuro Otras 85 millones sufren de diferentes formas de deterioro cognitivo y una pobre calidad de vida. La incidencia de EA para mayores de 60 aos es alrededor del 12%. En Chile hay 280.000 casos de EA. Los tratamientos son paliativos, con frmacos que tienen serios efectos adversos. No existen productos para su prevencin. Ineficacia de los medicamentos ha forzado a invertir altas sumas de dinero en I&D con

Alzheimers Disease A public health emergency.

Facts: Most common cause of dementia. Every year, 4.6 million new cases of dementia are reported worldwide. One new case every 7 seconds. By 2050, will be 100 million people with dementia in the world.
ADI, 2008

Total worldwide societal cost of Alzheimers disease (2005) US$ 315,4 billion

Latin America Total cost US$ 13.8 billions Total cost per demented US$ 6905
Wimo A, 2007

Epidemiologa
La enfermedad de Alzheimers causa el 60-80% of las demencias en mayores de 65 aos. 12% de las personas >65 tienen AD. 50% de las personas >85 tienen AD. 39.5 millones en el mundo tienen AD. AD genera un costo anual a la economa mundial sobre los USD 620 billones

Prdida de Neuronas Piramidales CDR 0.5 and 3.0

SMI-32 immunohistochemistry in layer IIIc of a CDR 0.5 case (A) and a CDR 3 case (B).
.

La hiptesis del amiloide llev a un camino equivocado que fren el avance en la solucin teraputica del Alzheimer
NORMAL
EXTRACELLULAR SOLUBLE APP (sAPP) BETA SECRETASE
ALPHA SECRETASE

sAPP
BETA-AMYLOID

BETAAMYLOID
NEURONAL MEMBRANE

AD

GAMMA SECRETASE

sAPP
CARBOXY TERMINUS INTRACELLULAR BETA-AMYLOID

Dekosky, Kaufer & Lopez. In: Neurology in Clinical Practice, 2004:1901-19

PROBLEMA A RESOLVER

Aug.172010 - 9:07 am |179 views|

Eli Lilly Alzheimers Drug Made Patients Worse

By MATTHEW Abti-Beta amyloid Patients taking an experimental Eli Lilly Alzheimers treatment worsened faster than those on placebo. The treatment also apparently caused skin cancer. Lilly says it will stop developing the drug, but will keep working on another, different Alzheimers treatment. The result is another setback for Alzheimer's research and should probably make scientists and investors think again about how little we know about this disease. From Lillyspress release: In two pivotal Phase III trials, semagacestat was compared with placebo in more than 2,600 patients with mild-to-moderate Alzheimers disease. Lilly has now reviewed data from a pre-planned interim analysis of semagacestat studies. This interim analysis showed that, as expected, cognition and the ability to complete activities of daily living of placebotreated patients worsened.

NUESTRA HIPOTESIS DE TAU

A: Protena Tau se une a los microtbulos. Sitios de fosforilacin e hiperfosforilacin, causa de la AD. B: Diagrama de flujo desde los cambios en tau a la formacin de los ovillos neurofibrilares y la demencia

Endogenous Damage Signals

External Factors

AGES

HMBG1 Deleterious S100 Head injury High fat Intake

Microglia
and astrocytes

RAGE

A peptide oligomers

Deficiency in B vitamins

TLR4 Infections NFk- ox-LDL Oxyradicals TNF IL-1 IL-6 Protective Plasma membrane Mechanical Damage Statins NSAIDs Long-term exposure to cholinergic agonists (?) Iron Overload

Degenerating neuron Cell cycle activation without proliferation Neuron Damage

Hyperphosphorylated tau protein

Maccioni et al, Arch Med. Res. 2001 Fernndez et al., J. Alz. Dis. 2008

Autotoxic diseases are different from autoimmune diseases

Autoimmune diseases involve the adaptive immune system. Aggressive and strike the young Autotoxic disease are less aggressive and strike the elderly. Much more prevalent than autoimmune diseases We postulate that the major driving force in AD is neuroinflammation. Activation of the innate immune system

Treatment with Antiinflammatory drugs reduces the risk of Alzheimer Disease control NSAIDS > 6 mos (.65) NSAIDS chronic (.5) NSAIDS >2y (.42) NSAIDS > 2y (.4)
NSAIDS > 2y (.2) Rheumatoid Arthritis (.16) 0.0 0.2 0.4 0.6

Yip et al 2005 Landi et al 2004 Zandi et al 2002 Stewart et al 1997 Int Veld et al 2001 McGeer et al 1990 0.8 1.0

Aggregated tau induces the release of proinflammatory cytokines IL-6 and TNF- . (Upper) Extracellular IL-6 in the conditioned media with ELISA protocol: (lower) ELISA of extracellular TNF- in GCM (A) Negative control buffer (B) 10 mg/mL aggregated tau (C ) 50 mg/mL aggregated tau

Exposure to activated glia conditionated media (GCM) for 24 h alter hippocampal neurons. A. Positive control with 1 mg/mL LPS. B. Exposure to conditioned media from glia treated with 50 g/mL aggregated tau. C. Exposure to conditioned media from glia treated with 10 g/mL aggregated tau and D. Negative control with aggregation buffer with arachidonic acid.

SIGNALING CASCADE EXPLAINING HOW IL-6 INDUCES TAU PHOSPHORYLATION Oxidative Stress NMDA

BetaAmyloid GLIA
IL6

IL-6 R
JAKs/ STATs

Neuron Ca2+

p38
MAPK

cdk5/p35

Tau-p

Egr-1
Quintanilla et al., 2005; Orellana et al., 2006, Maccioni et al., 2010

p35

 Compuesto Andino (Shilajit Andino): Es un producto orgnico natural derivado de la descomposicin milenaria de plantas encontradas en Los Andes chilenos. Su principio activo es el cido flvico, contiene adems cidos hmicos selenio y minerales.
23

As que por qu no mezclamos Shilajit Andino con Vitaminas del complejo B?

Publication of the Oxford Study on vitamins B in AD

VISTA AEROFOTOGRAMTRICA DEL REA MINERA CONTENIENDO LOS DEPSITOS DE SHILAJIT ANDINO EN LA REGIN DE ATACAMA.

LA REGION DEL ANDEAN COMPOUND

26

ICC GENERA BRAIN UP-10R ANDEAN COMPOUND PLUS VIT B COMPLEX Studies indicate that BrainUp-10: - Brain Up-10 is NEUROPROTECTOR - Brain Up-10 has neuritogenic activity - Brain Up-10 dos not have neurotoxicity (acute and chronic toxicity study) - Brain Up-10 no adverse effects in patients (Phase I). - Brain 10 improves cognitive performance (Clinical trials, Pilote

SOLUCION MEDICA
Tau antiaggregating activity
(A) TAU CONTROL (B) TAU + BRAINUP

(A) CONTROL

(B) + BRAIN UP

Neuritogenic action

Analisis morfomtrico de clulas del hipocampo expuestas a Brain Up-10

BRAIN UP-10R
Cornejo et al., J. Alz. Dis. 2011
Inhibitory effect of fulvic acid over PHFs formation as monitored by Thioflavin assay (ThT) . A. Aggregation of tau fragment 4RMBD in the presence of Fulvic Acid at different concentrations. The IC50 value for inhibitory effect of Fulvic Acid was 37 M. B. Suggested model for the Fulvic Acid structure

BRAIN UP-10R
Study of the changes in the blood biomarker for platelets tau in relation to the treatment with formulation
+ Formulation +Placebo Ratio of HMW/LMW (for explanation please see ***) Mean SD Mean Group of the clinical trial AD Patients* CONTROL** Normal SD Mean SD

Time Zero (0 weeks) Time 24 week

2.967+/- 0.368 2.320 +/- 0.541

2.879 +/- 0.567 + 3.369 +/- 0.697+

1.076 +/- 0.421 (1st. analysis) 1.107 +/- 0.239 (2nd. analysis)

Formulation group: 10 / N Placebo Group: 6 / N Control group: 7 * Patients incorporated into the study with pre diagnosis of mild to moderate AD and with incorporation criteria and informed consent signed and approved. ** Control subjects of the same range of ages of those incorporated as patients, from whom blood simples were extracted for the study to evaluate platelet tau. ***Ratio between oligomerized tau (HMW) and normal monomeric tau (LMW) (ratio of HMW/ LMW). This marker indicates that a significant increase in this ratio correlates with the level of cognitive impairment in AD. **** The blind was open. + Significant differences at the level of p<0.05

BRAIN UP-10R

En la evaluacin de sntomas neuropsiquitricos (Figura 4), destaca una clara tendencia a presentar menores alteraciones en el test NPI-12 en el grupo tratado con Brain-Up10R (valor p: 0,153).
Grupo
Placebo Brain Up10 100

Grupo
Placebo Brain Up10

50
80

40
60

Media NPI

30
40

NPI media

20

20

10
0

0
-20 0 12 24

12

24

Semana
Error bars: 95,00% CI

Semana
Error bars: 95,00% CI

Figura: Puntajes de los grupos placebo (azul) y tratado con Brain-Up10R (verde) en la evaluacin neuropsiquitrica mediante test NPI-12. Se aprecia un marcado efecto de Brain Up-10 R en aliviar los desordenes neuropsiquitricos del paciente, mayor estabilidad en la intensidad de los sntomas en los sujetos tratados con Brain Up-10R tanto en la evaluacin de frecuencia e intensidad de los sntomas (A), como en el grado de sufrimiento referido por el cuidador. Las barras de error corresponden a IC 95%.

BIOMARCADORES UN PUENTE ENTRE LA CIENCIA BASICA Y LA CLINICA

TAU BIOMARKERS IN CSF

Maccioni et al. (2006) Neurobiol Aging.

tau
Tau-P

Anomalous brain tau.

APP Variants

Tau-P

PROTEOMIC DETECTION

El Biomarcador Ideal
It should detect fundamental CNS pathophysiology of AD It should detect the presence of the disease itself, not the risk (e.g., APOE-4) It should be efficacious in pre-clinical stages It should be able to track disease progression, even from pre-clinical stages It should provide indication of treatment effectiveness It should be supported by clinico-pathological studies Better if it is non-invasive and inexpensive

Klunk WE. Neurobiology of Aging 1998; 19: 145-147

Niveles de (tau) en el CSF de pacientes con tres patologas neurolgicas

AD: Alzheimers disease VaD: Vascular dementia ALS: Amyotrophic Lateral Sclerosis FTD: Frontotemporal Dementia

*
pg/ml

** ***
(1)

Range: *345-951 pg/ml, **204-702 pg/ml, ***174-368 pg/ml. This study did not report S.D. and (1) did not include normal controls.

ESTRATEGIA BIOMARCADORES PARA DETECCION TEMPRANA AD Preclinical Phase

Tau Oligomers Biomarkers (e.g. Platelets tau. Potential early diagnosis) Deposits: Neurofibrilllary Tangles Decline of Instrumental Activities of Daily Living

Clinical Phase

(Diagnosis AD) Time (in years)

WWW.NEUROINOVATION.CL

METHODOLOGY
BIOMARKER

DIAGNOSIS TOOLS

VALIDATION CHILE (University Hospital)

VALIDATION USA (U. Pittsburgh)

PROTOTYPE FOR THE DIAGNOSIS KIT

DEVELOPMENT OF TECHNOLOGY

BIOMARCADOR NO INVASIVO DETECTA AD A NIVEL PRECLINICO

Refs: Neumann, Farias and Maccioni, 2010; Farias, Slachevsky and Maccioni, 2012

CLINICAL TRIALS OF AD PATIENTS IN SANTIAGO (n= 346) Team: Drs. A. Slachevsky, C. Delgado, P. Prez, G. Farias, L. Guzmn & R.B. Maccioni

Correlacion del progreso de la AD con el biomarcador de tau plaquetario: covariables edad & educacin p<0.001.

Lansoprazole Thioflavina S
50 m

LNS

THS

C
ThS

D
LNS

Real time study of the interaction of LNS with tau filaments

Covalently inmobilized protein Carboby methyldextran chains Gold surface (50 nm) Glass

Polymer PHF Amyloid

Inmobilization conditions Buffer pH 2.95, 3hrs Buffer pH 4.46, 3hrs

Biosensor

SPR Lansoprazole with AD-PHFs (violet) and with fibrillary A (green)

RU 4000 3500 3000 2500 2000 1500 1000 500 0 -500 -10

AD-PHF

20

50

80 Time (s)

110

140

170

200

Thanks / Gracias

Nuestros esfuerzos estan orientados a mejorar la calidad de vida

PROPUESTAS PARA ACTIVAR EL DESARROLLO DE NUEVAS TECNOLOGIAS Y SU TRANSFERENCIA

1.- Capital Humano. En el mundo desarrollado son cientficos con experiencia en gestin. 2.- Polticas de estado en C&T. Los gobiernos han estado concentrados en emular un modelo de comercializacin clsico y desincentivan a los cientficos a participar y emprender 3.- Incentivo a la generacin de productos. Los fondos solo permiten comprobar hiptesis y no su proyeccin a la generacin de productos empaquetables. 4.- Necesidad de spin-offs. Incentivos en las universidades para transferir tecnologas. 5.- Redes para conectar con centros en el resto del mundo y provocar un spill over de buenas practicas internacionales al sistema. 6.- Atraer empresas y centros tecnolgicos del mundo hacia Chile (no es fcil porque stos no confan en la institucionalidad actual)