Académique Documents
Professionnel Documents
Culture Documents
Drug screening
Sequence of experimentation and characterisation.
Objectives
Identify hits
Lead identification
Lead optimisation
Define the pharmacological and pharmacokinetic profile of the drug,
Methods
1. Non biological methods
2. Computational methods: Virtual screening, NMR
3. Biological methods
In vitro assays
Molecular assays
Cellular/Tissue/Organ assays
In-vivo assays
Systems or disease models
Biological methods
In- vitro assays
Molecular assays
Cellular/Tissue/Organ assays
In-vivo assays
Systems or disease models
1. Molecular assays
In-vitro assays
Detect the affinity and selectivity of the drug at the level of receptors.
May or may not detect the efficacy or the functional changes
Identification of the hit molecules(should beeffective at 50 mm conc).
E.g.
Cell membrane fractions from organs or cultured cells---source of receptors
----for affinity and selectivity
E.g.Alpha adrenoceptors to evaluate binding of alpha agonists
Enzymes e.g.Tyrosine hydroxylase from sympathetic nerve endings---enzyme
inhibition by –Inhibitors
Advantages
Large number of molecules can be screened as high throughput screening
Limitations
Functionalaspect cannot be evaluated
Other aspects like PK safety cannot be evaluated
2. Cellular assays
In vitro assays
Detect the cellular function-Affinity, selectivity and potency of the drug.
Effect on post-receptor mechanisms
Pharmacokinetics e.g.absorption across CaCo-2 cell lines
In-vitro metabolism Hep-G2 CELL lines
Hepatocytocytes and Kidney cell line-- toxicity
Drug- interaction studies
Lead identification
E.g.Islet cell culture—Beta cells for release of Insulin.
Hepatocyte culture for hepatotoxic effect
Advantages
More realistic models than the molecular assays.
Post target activation/or inhibition can be studied
Limitations
Cannot assess-
Dose determination
Effect on other organ systems
Tolerability
High throughput screening
Large libraries of chemicals are tested for their ability to modify the target
Most common target is the GPCR
Can screen upto 10,000 molecules per day
Ultra high throughput screening 1,00,000
Detects the agonists and antagonists
Also detects the selectivity of the drug using cross screening techniques.
Functional benefits
Automated sample generation and Preperation
Target finding and validation
Genomics
Proteomics
Parallel synthesis stations.etc.
HTS Advantages
Time saving
Large number can be screened
Techniques
Radioligand binding assays
Fluorescent assays
E.g.
Mouse or Rat models of anxiety for Benzodiazepine like drugs.