title: Virtual screening for LPA2-specific agonists identifies a nonlipid compound with antiapoptotic actions.
abstract: Lysophosphatidic acid (LPA) is a highly potent endogenous lipid mediator that protects and rescues cells from programmed cell death. Earlier work identified the LPA₂ G protein-coupled receptor subtype as an important molecular target of LPA mediating antiapoptotic signaling. Here we describe the results of a virtual screen using single-reference similarity searching that yielded compounds 2-((9-oxo-9H-fluoren-2-yl)carbamoyl)benzoic acid (NSC12404), 2-((3-(1,3-dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)propyl)thio)benzoic acid (GRI977143), 4,5-dichloro-2-((9-oxo-9H-fluoren-2-yl)carbamoyl)benzoic acid (H2L5547924), and 2-((9,10-dioxo-9,10-dihydroanthracen-2-yl)carbamoyl) benzoic acid (H2L5828102), novel nonlipid and drug-like compounds that are specific for the LPA₂ receptor subtype. We characterized the antiapoptotic action of one of these compounds, GRI977143, which was effective in reducing activation of caspases 3, 7, 8, and 9 and inhibited poly(ADP-ribose)polymerase 1 cleavage and DNA fragmentation in different extrinsic and intrinsic models of apoptosis in vitro. Furthermore, GRI977143 promoted carcinoma cell invasion of human umbilical vein endothelial cell monolayers and fibroblast proliferation. The antiapoptotic cellular signaling responses were present selectively in mouse embryonic fibroblast cells derived from LPA(1&2) double-knockout mice reconstituted with the LPA₂ receptor and were absent in vector-transduced control cells. GRI977143 was an effective stimulator of extracellular signal-regulated kinase 1/2 activation and promoted the assembly of a macromolecular signaling complex consisting of LPA₂, Na⁺ - H⁺ exchange regulatory factor 2, and thyroid receptor interacting protein 6, which has been shown previously to be a required step in LPA-induced antiapoptotic signaling. The present findings indicate that nonlipid LPA₂-specific agonists represent an excellent starting point for development of lead compounds with potential therapeutic utility for preventing the programmed cell death involved in many types of degenerative and inflammatory diseases.
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
authors.forename: Gyöngyi N
authors.initials: GN
authors.lastname: Kiss
authors.forename: James I
authors.initials: JI
authors.lastname: Fells
authors.forename: Renuka
authors.initials: R
authors.lastname: Gupte
authors.forename: Sue-Chin
authors.initials: SC
authors.lastname: Lee
authors.forename: Jianxiong
authors.initials: J
authors.lastname: Liu
authors.forename: Nóra
authors.initials: N
authors.lastname: Nusser
authors.forename: Keng G
authors.initials: KG
authors.lastname: Lim
authors.forename: Ramesh M
authors.initials: RM
authors.lastname: Ray
authors.forename: Fang-Tsyr
authors.initials: FT
authors.lastname: Lin
authors.forename: Abby L
authors.initials: AL
authors.lastname: Parrill
authors.forename: Balázs
authors.initials: B
authors.lastname: Sümegi
authors.forename: Duane D
authors.initials: DD
authors.lastname: Miller
authors.forename: Gabor
authors.initials: G
authors.lastname: Tigyi
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D016466:CHO Cells; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D053938:DNA Fragmentation / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D060588:LIM Domain Proteins / Q000378:metabolism; D008246:Lysophospholipids / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D018345:Mice, Knockout; D009361:Neoplasm Invasiveness; D010750:Phosphoproteins / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D049368:Receptors, Lysophosphatidic Acid / Q000819:agonists* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents; D060588:LIM Domain Proteins; D008246:Lysophospholipids; C092282:PSMC5 protein, human; D010750:Phosphoproteins; D043562:Receptors, G-Protein-Coupled; D049368:Receptors, Lysophosphatidic Acid; D017923:Sodium-Hydrogen Exchangers; D014157:Transcription Factors; D051028:bcl-2-Associated X Protein; C093376:sodium-hydrogen exchanger regulatory factor; D011065:Poly(ADP-ribose) Polymerases; D020169:Caspases; D046988:Proteasome Endopeptidase Complex; D000074183:ATPases Associated with Diverse Cellular Activities; C032881:lysophosphatidic acid