title: Weak affinity chromatography as a new approach for fragment screening in drug discovery.
abstract: Fragment-based drug design (FBDD) is currently being implemented in drug discovery, creating a demand for developing efficient techniques for fragment screening. Due to the intrinsic weak or transient binding of fragments (mM-μM in dissociation constant (K(D))) to targets, methods must be sensitive enough to accurately detect and quantify an interaction. This study presents weak affinity chromatography (WAC) as an alternative tool for screening of small fragments. The technology was demonstrated by screening of a selected 23-compound fragment collection of documented binders, mostly amidines, using trypsin and thrombin as model target protease proteins. WAC was proven to be a sensitive, robust, and reproducible technique that also provides information about affinity of a fragment in the range of 1 mM-10 μM. Furthermore, it has potential for high throughput as was evidenced by analyzing mixtures in the range of 10 substances by WAC-MS. The accessibility and flexibility of the technology were shown as fragment screening can be performed on standard HPLC equipment. The technology can further be miniaturized and adapted to the requirements of affinity ranges of the fragment library. All these features of WAC make it a potential method in drug discovery for fragment screening.
authors.affiliation: School of Natural Sciences, Linnaeus University, Kalmar, Sweden.
authors.forename: Minh-Dao
authors.initials: MD
authors.lastname: Duong-Thi
authors.forename: Elinor
authors.initials: E
authors.lastname: Meiby
authors.forename: Maria
authors.initials: M
authors.lastname: Bergström
authors.forename: Tomas
authors.initials: T
authors.lastname: Fex
authors.forename: Roland
authors.initials: R
authors.lastname: Isaksson
authors.forename: Sten
authors.initials: S
authors.lastname: Ohlson
mesh_terms: D000578:Amidines / Q000737:chemistry* / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D055808:Drug Discovery / Q000379:methods*; D006801:Humans; D054852:Small Molecule Libraries; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000737:chemistry* / Q000494:pharmacology*
chemical_list: D000578:Amidines; D054852:Small Molecule Libraries; D014361:Trypsin Inhibitors; D014357:Trypsin; D013917:Thrombin