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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.48 [2011] >
IJBB Vol.48(4) [August 2011] >
| Title: | High-throughput screening assays for cyclooxygenase-2 and 5-lipoxygenase, the targets for inflammatory disorders |
| Authors: | Kumar, K Anil
Reddy, T Chandramohan
Reddy, Gorla V
Reddy, D Bharat Kumar
Mahipal, S V K
Sinha, Sudhir
Gaikwad, Anil N
Reddanna, P |
| Keywords: | High-throughput screening
Cyclooxygenase
Lipoxygenase
NSAIDs
COXIBs
CLOXIBs |
| Issue Date: | Aug-2011 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | High-throughput screening (HTS) involves testing of compound libraries
against validated drug targets using quantitative bioassays to identify ‘hit’
molecules that modulate the activity of target, which forms the starting point
of a drug discovery effort. Eicosanoids formed via cyclooxygenase (COX) and
lipoxygenase (LOX) pathways are major players in
various inflammatory disorders. As the conventional non-steroidal
anti-inflammatory drugs (NSAIDs) that inhibit both the constitutive (COX-1) and
the inducible (COX-2) isoforms have gastric and renal side effects and the
recently developed COX-2 selective anti-inflammatory drugs (COXIBs) have
cardiac side effects, efforts are being made to develop more potent and safer
anti-inflammatory drugs. Current assay
methods for these enzymes, such as oxygraphic, radioisotopic,
spectrophotometric etc. are not compatible for screening of large number of
compounds as in drug discovery programs. In the
present study, HTS-compatible assays for COX-1, COX-2 and 5-LOX were developed for screening
of compound libraries with the view to identify potential anti-inflammatory
drug candidates. A spectrophotometric assay involving co-oxidation of
tetramethyl-p-phenylene diamine (TMPD) during the reduction of prostaglandin G2
(PGG2) to PGH2 was adopted and standardized for screening
of compounds against COX-1 and COX-2. Similarly, the HTS-compatible FOX
(ferrous oxidation-xylenol orange) based spectrophotometric assay involving the
formation of Fe3+/xylenol orange complex showing absorption in the
visible range was developed for screening of compounds against 5-LOX. |
| Page(s): | 256-261 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0959 (Online); 0301-1208 (Print) |
| Source: | IJBB Vol.48(4) [August 2011]
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