Biomarkers in drug discovery and development · aptuit.com aptuit.com. Biomarkers are used in pre-...
Transcript of Biomarkers in drug discovery and development · aptuit.com aptuit.com. Biomarkers are used in pre-...
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Margit Wissenbach,PhD
Dir Business Development Scandinavia
Biomarkers in drug discoveryand development
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Integrated Discovery Integrated Development
INDCandidateTo specification
Aptuit: A Truly Integrated R&D Offering
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Biomarkers are used in pre-clinical and clinical drug development as indicators of likely response to drug: therefore, they enjoy utility both as predictors of possible EFFICACY and
also as warning signs of potential TOXICITY
Efficacy biomarkers
rapid confirmation that the drug has hit its
targetIndicator of
pharmacological activity
Safety biomarkers
measurable indicators of toxicological and adverse side effects
of drug activitycells
proteins
genes
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Quality assessment of biological drug activity Translational approach to clinic development
Reduced attrition
Biomarkers
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Applications for biomarkers
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Single biomarker approach (ELISA, EIA, RIA, IRMA);
Disease-driven biomarker panels (Luminex® and Meso Scale Discovery® platforms):
acute stress markers: corticosterone and ACTH - potential indicators of specific compound activity or side effects;
metabolic markers/adipokines: prolactin, insulin, amylin, glucagon, GLP-1, glucose, leptin, resistin, adiponectin, MCP-1, ghrelin - to assess metabolic liability displayed by some compounds;
inflammatory markers: Cytokines/Chemokines (e.g. IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, GRO-KC, MIP-1α, GM-CSF, IFN-γ & TNF-α) to test for pro- or anti-inflammatory activity;
hypothalamic neuroendocrine markers: prolactin, TSH, LH, GH, FSH; neuroplasticity panel: serum BDNF, VEGF, cAMP; sex hormones: testosterone, progesterone, estradiol.
Quantification in complex biological samples (e.g. serum, plasma, urine, saliva, brain tissue) from preclinical species and in clinical samples
Use of biomarkers in support of disease hypothesis
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Regulatory requirements for biomarker validation
Little regulatory guidance on biomarker assay validation is available: In 2003 the American Association of Pharmaceutical Scientists participated in
a Biomarker Method Validation Workshop to address the validation challenges of biomarker assays in support of drug development Biomarker method development and validation is based on a “fit-for-
purpose” approach.
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Cardiac Troponin I (cTnI):
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represents a biomarker of cardiac toxicity in the clinic
can be used as reverse translational biomarker since animal studies have demonstrated that it’s a marker of the same diseases that occur in humans
has been successfully applied as marker of cardiac injury in different preclinical animal species
is measured in preclinical by automated chemiluminescent immunoassay on the Siemens ADVIA Centaur CP
courtesy by jolisfukyu.tokai-sc.jaea.go.jp
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Aminoterminal pro-brain natriuretic peptide (NT-proBNP):
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courtesy by Roche Diagnostics
is primarily secreted from the cardiac ventricular myocytes in response to cardiac stress
used in the clinic for the diagnosis of left ventricular systolic and diastolic dysfunction
has been evaluated in preclinical studies in the dog
is measured in preclinical by ELISA
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Biomarkers in a 26-week dog study
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CTnI was essential in the characterization of the onset of the lesions and its slow evolution;
excellent correlation between cTnI and developing pathology was observed with cTnIelevations detectable long before light microscopy changes;
transmission electron microscopy analysis was capable to detect ultrastructural changes in the heart which were concomitant to cTnI elevations
NT-proBNP determination in conjunction with echocardiography and morphometry acted as an early marker of heart hypertrophy anticipating echocardiography changes as well morphometry
cTnI and NT-proBNP are reliable, sensitive and non-invasive markers of drug-induced cardiac changes in the dog
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CARDIAC MARKERS NT-proBrain Natriuretic Peptide Serum ELISA Atrial Natriuretic Peptide Serum ELISA Cardiac Troponine I Serum MSD or Centaur CP
Immunoassay Cardiac Troponin T Serum MSD Fatty acid binding protein 3 Serum MSD Myosin light chain 3 Serum MSD CK-MB Serum ELISA AST, myoglobin, aldolase, CK and LDH isoemzymes Serum Clinical Chemistry
Analyzer HORMONES Prolactin Serum Immulite 2000
Immunoassay Growth hormone Serum Immulite 2000
Immunoassay Insulin Serum Immulite 2000
Immunoassay IGF-1 (Insulin-like growth factor 1) Serum ELISA INFLAMMATION/IMMUNITY IL-1beta, TNF-alpha, IFN-gamma, IL-2, IL-6, IL-12/23 (p40)
Plasma Luminex
IFN-gamma Splenocytes ELISpot Intracellular cytokines Whole
blood/tissues Flow Cytometry
Histamine Serum ELISA T cell activation markers (CD25,CD69, HLA-DR) Whole blood,
PBMC Flow cytometry
T/B/NK cells Immunophenotyping Whole blood Flow cytometry LIVER INJURY MARKERS Arginase 1 Arginase 1 Arginase 1 αGST αGST αGST
CARDIAC MARKERS
NT-proBrain Natriuretic Peptide
Serum
ELISA
Atrial Natriuretic Peptide
Serum
ELISA
Cardiac Troponine I
Serum
MSD or Centaur CP Immunoassay
Cardiac Troponin T
Serum
MSD
Fatty acid binding protein 3
Serum
MSD
Myosin light chain 3
Serum
MSD
CK-MB
Serum
ELISA
AST, myoglobin, aldolase, CK and LDH isoemzymes
Serum
Clinical Chemistry Analyzer
HORMONES
Prolactin
Serum
Immulite 2000 Immunoassay
Growth hormone
Serum
Immulite 2000 Immunoassay
Insulin
Serum
Immulite 2000 Immunoassay
IGF-1 (Insulin-like growth factor 1)
Serum
ELISA
INFLAMMATION/IMMUNITY
IL-1beta, TNF-alpha, IFN-gamma, IL-2, IL-6, IL-12/23 (p40)
Plasma
Luminex
IFN-gamma
Splenocytes
ELISpot
Intracellular cytokines
Whole blood/tissues
Flow Cytometry
Histamine
Serum
ELISA
T cell activation markers (CD25,CD69, HLA-DR)
Whole blood, PBMC
Flow cytometry
T/B/NK cells Immunophenotyping
Whole blood
Flow cytometry
LIVER INJURY MARKERS
Arginase 1
Arginase 1
Arginase 1
αGST
αGST
αGST
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ONCOLOGY AlPhafetoprotein (AFP) Serum ELISA Caspase3 Tumor section Immunohistochemistry Carbohydrate antigen-19-9 (CA-19-9) Serum ELISA Carbohydrate antigen-15-3 (CA-15-3) Serum ELISA Carbohydrate antigen-125 (CA-125) Serum ELISA Carcinogenic Embryonic Antigen (CEA) Serum Chemiluminescent
Immunoassay CD31 Tumor section Immunohistochemistry CYFRA 21-1 (Cytokeratin) Serum Monoclonal based assay Galectin-3 Serum ELISA HER2 Expression Serum ELISA Ki67 Tumor section Immunohistochemistry PCNA Tumor section Immunohistochemistry p-histone-3 Tumor section Immunohistochemistry Prostate Specific Antigen (PSA) Blood/plasma Chemiluminescent
Immunoassay Putresceine TPA (Cytokeratin) Serum Monoclonal based assay TPS (Cytokeratin) Serum Monoclonal based assay TRAIL/TRAIL Rs Blood/plasma TUNEL Tumor section Immunohistochemistry VEGF, VEGF-C, VEGF-D, Tie-2, Flt-1, PIGF and bFGF Serum MSD P53 Serum MSD OXIDATIVE STRESS MARKERS GSH-GSSG ratio Muscle and liver Fluorimetric Assay Nitrotyrosine Muscleand liver ELISA Reactive Oxygen Species (ROS) Plasma, muscle
and liver Fluorimetric Assay
ONCOLOGY
AlPhafetoprotein (AFP)
Serum
ELISA
Caspase3
Tumor section
Immunohistochemistry
Carbohydrate antigen-19-9 (CA-19-9)
Serum
ELISA
Carbohydrate antigen-15-3 (CA-15-3)
Serum
ELISA
Carbohydrate antigen-125 (CA-125)
Serum
ELISA
Carcinogenic Embryonic Antigen (CEA)
Serum
Chemiluminescent Immunoassay
CD31
Tumor section
Immunohistochemistry
CYFRA 21-1 (Cytokeratin)
Serum
Monoclonal based assay
Galectin-3
Serum
ELISA
HER2 Expression
Serum
ELISA
Ki67
Tumor section
Immunohistochemistry
PCNA
Tumor section
Immunohistochemistry
p-histone-3
Tumor section
Immunohistochemistry
Prostate Specific Antigen (PSA)
Blood/plasma
Chemiluminescent Immunoassay
Putresceine
TPA (Cytokeratin)
Serum
Monoclonal based assay
TPS (Cytokeratin)
Serum
Monoclonal based assay
TRAIL/TRAIL Rs
Blood/plasma
TUNEL
Tumor section
Immunohistochemistry
VEGF, VEGF-C, VEGF-D, Tie-2, Flt-1, PIGF and bFGF
Serum
MSD
P53
Serum
MSD
OXIDATIVE STRESS MARKERS
GSH-GSSG ratio
Muscle and liver
Fluorimetric Assay
Nitrotyrosine
Muscleand liver
ELISA
Reactive Oxygen Species (ROS)
Plasma, muscle and liver
Fluorimetric Assay
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RENAL MARKERS Albumine Serum and urine MSD Lipocalin -2 Serum and urine MSD KIM-! Serum and urine MSD Osteopontin Serum and urine MSD SEX HORMONES Progesterone Serum ELISA Testosterone Serum ELISA Dihydrotestosterone Serum ELISA STRESS MARKERS Corticosterone Serum/plasma RIA THYROID MARKERS Triiodothyronine (T3) Serum Centaur CP Immunoassay
or ELISA Thyroxine (T4) Serum Centaur CP Immunoassay
or ELISA Thyroid stimulating hormone (TSH) Serum Centaur CP Immunoassay
or ELISA or Immulite 2000 Immunoassay
RENAL MARKERS
Albumine
Serum and urine
MSD
Lipocalin -2
Serum and urine
MSD
KIM-!
Serum and urine
MSD
Osteopontin
Serum and urine
MSD
SEX HORMONES
Progesterone
Serum
ELISA
Testosterone
Serum
ELISA
Dihydrotestosterone
Serum
ELISA
STRESS MARKERS
Corticosterone
Serum/plasma
RIA
THYROID MARKERS
Triiodothyronine (T3)
Serum
Centaur CP Immunoassay or ELISA
Thyroxine (T4)
Serum
Centaur CP Immunoassay or ELISA
Thyroid stimulating hormone (TSH)
Serum
Centaur CP Immunoassay or ELISA or Immulite 2000 Immunoassay
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InstrumentationTECHNOLOGIES
LuminexECL
ELISA
Flow Cytometry
ELIspot
MATRIX
Rodents blood and derivativesPrimate blood and derivativesHuman blood and derivatives
Primary cells
Gene expression
Biomarkers in drug discovery�and development Aptuit: A Truly Integrated R&D Offering BiomarkersApplications for biomarkersUse of biomarkers in support of disease hypothesis�Regulatory requirements for biomarker validationCardiac Troponin I (cTnI):Aminoterminal pro-brain natriuretic peptide (NT-proBNP):Biomarkers in a 26-week dog study Dias nummer 10Dias nummer 11Dias nummer 12Instrumentation