Hit Identification

The entry point for chemistry program within drug discovery research is
generally the identification of molecules of high specificity with an
adequate activity in a suitable target assay. Such initial hits can be
generated in a number of ways. It is therefore important to employ
alternative hit-identification strategies that are able to tackle a variety
of biological macromolecular targets effectively, and to identify
proprietary, synthetically tractable and pharmacologically relevant
compounds rapidly. These methods can be subdivided into those that
require very detailed ligand and/or target information, and those that do
not. The former include techniques such as nuclear magnetic
resonance (NMR) and X-ray crystallography. Those strategies that do
not require any prior information on target or ligand are using
serendipity-based search strategies in either a given physical or virtual
compound subset. Examples of random hit-identification strategies can
be included by biophysical and biochemical testing that generally
employ the method of detecting a molecular-binding event, usually in
a high-throughput screening (HTS).

BOC Sciences provides varieties of methods including NMR, X-ray

crystallography and HTS for hit identification.

 NMR

NMR is a physical phenomenon, and by studying the peaks of NMR

spectra, the structure of compounds can be determined. It can be a very
selective technique to distinguish atoms in a molecule or molecules of
the same type according to their local chemical conditions. As a useful
tool in the hit identification, NMR spectroscopy techniques have been
widely applied to detect ligand binding and monitor enzyme kinetics and
inhibition. For example, methods for detecting binding by NMR fall into
two main categories: those monitoring NMR signals from the protein,
such as protein chemical-shift perturbation studies, and those
monitoring the ligand, which exploit the large differences in the rates of
 rotational and translational motions of a small molecule in the free state
relative to when it is bound to a macromolecule. A common protein-
based approach is to label the target receptor with stable isotopes such
as 15N and/or 13C. Binding of a ligand or a macromolecule alters the
chemical environment around the binding site, which will perturb the
chemical shift of magnetic nuclei at this site.

 X-ray crystallography

X-ray crystallography is a technique developed to analyze the atomic

and molecular structures of crystals by determining the X-rays caused
by the crystalline atoms. By measuring the angle and the intensity of
these beams, a three-dimensional picture of the electron density within
a crystal is given. For example, protein X-ray crystallography can be
applied to the rationalization of the fragment-binding mode in the ATP
binding site of Hsp90, which is an important member of a class of
proteins known as molecular chaperones. This structural information is
then used to evolve the fragments to hits, either by growing the
fragment or combining structural features from a lot of compounds.

 HTS

HTS is a scientific method widely used in drug discovery, which can

help identify hits, antibodies, or genes that regulate a specific biological
pathway. The results of HTS will provide identification of hit as a starting
point for drug design and an understanding of the interactions or roles
in specific biochemical processes in drug discovery.

BOC Sciences has a series of methods that can provide high-quality

and effective hit compounds for your drug discovery. We would like to
hear from you and look forward to cooperating with you.

Reference

1. Napolitano, C. (2009) “DESIGN AND SYNTHESIS OF NOVEL SCAFFOLDS

AND BUILDING BLOCKS (Doctoral dissertation, Università degli studi di
Ferrara)’.