Fluorescent HTS Assays for Methyltransferases in Neurodegenerative Diseases

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$822,148.00
Award Year:
2008
Program:
SBIR
Phase:
Phase II
Contract:
2R44GM073290-02
Agency Tracking Number:
GM073290
Solicitation Year:
2008
Solicitation Topic Code:
n/a
Solicitation Number:
PHS2007-2
Small Business Information
BELLBROOK LABS, LLC
5500 Nobel Drive, Suite 250, MADISON, WI, 53711
Hubzone Owned:
Y
Socially and Economically Disadvantaged:
Y
Woman Owned:
Y
Duns:
119165251
Principal Investigator:
ROBERT LOWERY
(608) 227-4501
BOB.LOWERY@BELLBROOK-LABS.COM
Business Contact:
(608) 227-4501
bob.lowery@bellbrooklabs.com
Research Institution:
n/a
Abstract
DESCRIPTION (provided by applicant): Methylation is a ubiquitous and reversible covalent modification used to control the function of diverse biomolecules including their activity, stability and localization. Like phosphorylation, methylation is used to mo dulate protein function, but in addition many small molecules are regulated by methylation including hormones, neurotransmitters, xenobiotics, and lipids. More than 50 distinct methyltransferase (MT's) enzymes are present in humans, and not surprisingly th ey are emerging as therapeutic targets for a broad range of diseases. Their involvement in neurodegenerative disease pathways is of special relevance because of the central role that methylation plays in regulating neurotransmitter function. The developmen t of highly selective MT modulators is clearly a compelling medical priority. However, efforts to achieve this are being hampered by a lack of robust, flexible enzyme assay methods adaptable to high throughput screening (HTS). To address this need, in Phas e I BellBrook Labs developed a novel fluorescence polarization immunoassay for S-adenosylhomocysteine (SAH), the invariant product of all MT reactions. This required development of a highly selective antibody that is able to differentiate between SAH and t he substrate S-adenosylmethionine on the basis of a single methyl group. The Transcreener MT assay enables detection of any MT with any acceptor substrate in a homogenous format - and without the use of coupling enzymes - thus it eliminates all of the shor tcomings associated with alternative methods. In Phase II we propose to 1) rapidly commercialize a first generation, fluorescence polarization-based Transcreener MT assay based of the polyclonal antibodies developed in Phase I, 2) increase flexibility and ease-of-use for the MT assay by using targeted mutagenesis of recombinant IgG clones to increase antibody selectivity for SAH, 3) increase HTS market penetration by formatting the assay for a second commonly used fluorescent detection mode, TR-FRET, and 4) use the novel MT assay combined with in silico screening to develop isoform selective inhibitors for five small molecule methyltransferases. The availability of robust, generic fluorescent HTS assay methods and selective chemical probes for MT's should gr eatly accelerate their validation as drug targets and movement into the pharma drug discovery pipeline.

* information listed above is at the time of submission.

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