A Pyrazole Derivative, YM-58483, Potently Inhibits Store-Operated Sustained Ca2+ Influx and IL-2 Production in T Lymphocytes

Ishikawa, Jun; Ohga, Keiko; Yoshino, Taiji; Takezawa, Ryuichi; Ichikawa, Atsushi; Kubota, Hirokazu; Yamada, Toshimitsu

doi:10.4049/jimmunol.170.9.4441

Cited by 237 publications

(198 citation statements)

References 30 publications

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“…This contradicts with the report that Pyr2 failed to alter membrane potential in Jurkat cells (25). In addition, when external Na ϩ ions were completely replaced by the nonpermeant organic cation N-methyl-Dglucamine, Sr 2ϩ influx activated by CCh still showed an intact sensitivity to Pyr2, leading to an idea that the action of Pyr2 to block TRPC3 channels is independent of the membranedepolarizing action of TRPM4 (27).…”

Section: Discussioncontrasting

confidence: 53%

“…Several reports have suggested that BTP2 (Pyr2) is a potent inhibitor for both Ca 2ϩ release-activated Ca 2ϩ (CRAC) channels and TRPC channels and for NFAT-driven IL-2 production (25)(26)(27). Structure-function relationships in BTPs proposed that 4Ј-[3,5-bis(trifluoromethyl)pyrazol-1-yl]-carboxanilide moiety is useful for discovering potent inhibitors for CRAC channels (37).…”

Section: Discussionmentioning

confidence: 99%

“…The BTP derivative 4-methyl-4Ј-[3,5-bis(trif luoromethyl)-1H-pyrazol-1-yl]-1,2,3-thiadiazole-5-carboxanilide, BTP2 (YM-58483), was shown to block SOCs in T lymphocytes and TRPC channels in HEK293 cells (25)(26)(27). Importantly, unlike other TRP inhibitors, SK&F 96365 and 2-aminoethyldiphenylborate, BTP2 is selective to TRP channels and does not affect Ca 2ϩ handling by mitochondria or ER, or K ϩ channels or voltage-dependent Ca 2ϩ channels (25)(26)(27). Pharmacological profiles of BTP2 have been investigated in vitro and in vivo to evaluate its potential as a therapeutic anti-asthma drug (28).…”

Section: Canonical Transient Receptor Potential (Trpc) Channels Contrmentioning

confidence: 99%

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Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound

Kiyonaka

Kato

Nishida

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

336

318

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Canonical transient receptor potential (TRPC) channels control influxes of Ca 2؉ and other cations that induce diverse cellular processes upon stimulation of plasma membrane receptors coupled to phospholipase C (PLC). Invention of subtype-specific inhibitors for TRPCs is crucial for distinction of respective TRPC channels that play particular physiological roles in native systems. Here, we identify a pyrazole compound (Pyr3), which selectively inhibits TRPC3 channels. Structure-function relationship studies of pyrazole compounds showed that the trichloroacrylic amide group is important for the TRPC3 selectivity of Pyr3. Electrophysiological and photoaffinity labeling experiments reveal a direct action of Pyr3 on the TRPC3 protein. In DT40 B lymphocytes, Pyr3 potently eliminated the Ca 2؉ influx-dependent PLC translocation to the plasma membrane and late oscillatory phase of B cell receptorinduced Ca 2؉ response. Moreover, Pyr3 attenuated activation of nuclear factor of activated T cells, a Ca 2؉ -dependent transcription factor, and hypertrophic growth in rat neonatal cardiomyocytes, and in vivo pressure overload-induced cardiac hypertrophy in mice. These findings on important roles of native TRPC3 channels are strikingly consistent with previous genetic studies. Thus, the TRPC3-selective inhibitor Pyr3 is a powerful tool to study in vivo function of TRPC3, suggesting a pharmaceutical potential of Pyr3 in treatments of TRPC3-related diseases such as cardiac hypertrophy.Ca 2ϩ signaling ͉ pyrazole compounds ͉ TRPC channels ͉ TRPC3

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Section: Discussioncontrasting

confidence: 53%

Section: Discussionmentioning

confidence: 99%

Section: Canonical Transient Receptor Potential (Trpc) Channels Contrmentioning

confidence: 99%

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Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound

Kiyonaka

Kato

Nishida

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

336

318

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“…Consequently, CRAC channels have been proposed as potential target in the therapeutic management of autoimmune and inflammatory disorders. Recent in vitro studies showed that pyrazole derivatives potently inhibit Ca 2ϩ influx through CRAC, thus counteracting T cell activation and NFAT-driven IL-2 production by activated T cells (21,22).…”

Section: D4mentioning

confidence: 99%

Targeting Gut T Cell Ca2+ Release-Activated Ca2+ Channels Inhibits T Cell Cytokine Production and T-Box Transcription Factor T-Bet in Inflammatory Bowel Disease

Sabatino

Rovedatti

Kaur

et al. 2009

The Journal of Immunology

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Prolonged Ca2+ entry through Ca2+ release-activated Ca2+ (CRAC) channels is crucial in activating the Ca2+-sensitive transcription factor NFAT, which is responsible for directing T cell proliferation and cytokine gene expression. To establish whether targeting CRAC might counteract intestinal inflammation, we evaluated the in vitro effect of a selective CRAC inhibitor on T cell cytokine production and T-bet expression by lamina propria mononuclear cells (LPMC) and biopsy specimens from inflammatory bowel disease (IBD) patients. The inhibitory activity of the CRAC blocker was investigated through patch-clamp experiments on rat basophilic leukemia cells and fluorometric imaging plate reader intracellular Ca2+ assays using thapsigargin-stimulated Jurkat T cells and its detailed selectivity profile defined using a range of in vitro radioligand binding and functional assays. Anti-CD3/CD28-stimulated LPMC and biopsy specimens from 51 patients with IBD were cultured with a range of CRAC inhibitor concentrations (0.01–10 μM). IFN-γ, IL-2, IL-8, and IL-17 were analyzed by ELISA. T-bet was determined by immunoblotting. We found that the CRAC blocker concentration-dependently inhibited CRAC current in rat basophilic leukemia cells and thapsigargin-induced Ca2+ influx in Jurkat T cells. A concentration-dependent reduction in T-bet expression and production of IFN-γ, IL-2, IL-17, but not IL-8, was observed in IBD LPMC and biopsy specimens treated with the CRAC inhibitor. In conclusion, we provide evidence that the suppression of CRAC channel function may dampen the increased T cell response in the inflamed gut, thus suggesting a promising role for CRAC inhibitor drugs in the therapeutic management of patients with IBD.

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“…Specific www.nature.com/aps Zhang HZ et al Acta Pharmacologica Sinica npg CRAC channel inhibitors may be developed into new safe and potent immune suppressors, which would benefit patients with these diseases. Thus far, a variety of small molecules blocking the CRAC channel have been identified, such as the imidazole derivative, SKF96365 (IC 50 of approximately 4 μmol/L) [15,16] , 2-APB [at low concentrations of 1-5 μmol/L, 2-APB potentiates CRAC currents, whereas high concentrations (>10 μmol/L) cause a transient enhancement of CRAC currents followed by a complete block in S2 for mammalian cells] [17,18] , YM58483 (IC 50 of approximately 150 nmol/L after 24 h of preincubation; IC 50 of approximately 10 μmol/L immediately after addi tion) [19,20] , and Synta 66 (IC 50 of approximately 3 μmol/L) [21] ( Figure 1); however, these compounds are not specific because they interfere with a variety of other transport processes. The only specific CRAC channel inhibitor tested in human is CM2489 [22] , the structure of which has not yet been disclosed.…”

Section: Introductionmentioning

confidence: 99%

Discovery and structural optimization of 1-phenyl-3-(1-phenylethyl)urea derivatives as novel inhibitors of CRAC channel

Zhang

Chen

et al. 2015

Acta Pharmacol Sin

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Aim: Ca 2+ -release-activated Ca 2+ (CRAC) channel, a subfamily of store-operated channels, is formed by calcium release-activated calcium modulator 1 (ORAI1), and gated by stromal interaction molecule 1 (STIM1). CRAC channel may be a novel target for the treatment of immune disorders and allergy. The aim of this study was to identify novel small molecule CRAC channel inhibitors. Methods: HEK293 cells stably co-expressing both ORAI1 and STIM1 were used for high-throughput screening. A hit, 1-phenyl-3-(1-phenylethyl)urea, was identified that inhibited CRAC channels by targeting ORAI1. Five series of its derivatives were designed and synthesized, and their primary structure-activity relationships (SARs) were analyzed. All derivatives were assessed for their effects on Ca 2+ influx through CRAC channels on HEK293 cells, cytotoxicity in Jurkat cells, and IL-2 production in Jurkat cells expressing ORAI1-SS-eGFP. Results: A total of 19 hits were discovered in libraries containing 32 000 compounds using the high-throughput screening. 1-Phenyl-3-(1-phenylethyl)urea inhibited Ca 2+ influx with IC 50 of 3.25±0.17 μmol/L. SAR study on its derivatives showed that the alkyl substituent on the α-position of the left-side benzylic amine (R1) was essential for Ca 2+ influx inhibition and that the S-configuration was better than the R-configuration. The derivatives in which the right-side R3 was substituted by an electron-donating group showed more potent inhibitory activity than those that were substituted by electron-withdrawing groups. Furthermore, the free N-H of urea was not necessary to maintain the high potency of Ca 2+ influx inhibition. The N,N′-disubstituted or N′-substituted derivatives showed relatively low cytotoxicity but maintained the ability to inhibit IL-2 production. Among them, compound 5b showed an improved inhibition of IL-2 production and low cytotoxicity. Conclusion: 1-Phenyl-3-(1-phenylethyl)urea is a novel CRAC channel inhibitor that specifically targets ORAI1. This study provides a new chemical scaffold for design and development of CRAC channel inhibitors with improved Ca 2+ influx inhibition, immune inhibition and low cytotoxicity.

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A Pyrazole Derivative, YM-58483, Potently Inhibits Store-Operated Sustained Ca2+ Influx and IL-2 Production in T Lymphocytes

Cited by 237 publications

References 30 publications

Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound

Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound

Targeting Gut T Cell Ca2+ Release-Activated Ca2+ Channels Inhibits T Cell Cytokine Production and T-Box Transcription Factor T-Bet in Inflammatory Bowel Disease

Discovery and structural optimization of 1-phenyl-3-(1-phenylethyl)urea derivatives as novel inhibitors of CRAC channel

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