selective par1 antagonist  (MedChemExpress)

Journal: PLoS ONE

Article Title: Mechanisms of thrombin-Induced myometrial contractions: Potential targets of progesterone

doi: 10.1371/journal.pone.0231944

Figure Lengend Snippet: (A-C) Immunofluorescence of PAR1 (green) in myometrium from a pregnant woman (A) , non-pregnant woman ( B ), and fetal membrane (C) . Nuclei were stained with DAPI (blue). Am, amnion, Cho, chorion, Deci, decidua. (D-G) Localization of hemorrhage, thrombin, and PAR1 in placental abruption at 25 weeks of gestation (D and E) , and 33 weeks gestation (F and G) resulting in disseminated intravascular coagulopathy and uterine bleeding requiring hysterectomy for hemostasis. (D and F) Hematoxylin and eosin staining of the myometrium adjacent to the placenta. Note that hemorrhage infiltrated the myometrium. Bars, 50 μm. (E and G) Immunofluorescence of PAR1 (green), thrombin (red), and DAPI (blue) at the same location of (A) . Bars, 50 μm.

Article Snippet: On the day of experiments, cells in collagen gels were pretreated with the following reagents for 1 h: 100 nM PAR1 selective antagonist SCH79797 (No.1592 Tocris), 1 μM ROCK inhibitor Y-27632 (10005583, Cayman Chemical), 10 μM MLCK inhibitor, ML-7 (11801, Cayman Chemical), 10 μM of indomethacin (I-7378, Sigma), or 1 μM of progesterone (28921–64, Nacalai-tesque).

Techniques: Immunofluorescence, Membrane, Staining

Journal: PLoS ONE

Article Title: Mechanisms of thrombin-Induced myometrial contractions: Potential targets of progesterone

doi: 10.1371/journal.pone.0231944

Figure Lengend Snippet: Thrombin increased contraction of primary human myometrial cells through PAR1. (A and B) (Left images) Representative images of collagen lattice assay of human myometrial cells at 30 min treated with PBS (Ctl) and thrombin (A) or PAR1 activating peptide, TFLLR (B) . (Right graphs) Quantification of myometrial contractions in collagen lattice assays (n = 3). **, p < 0.01 at each time point. (C) Collagen lattice assay of myometrial cells at 30 min with 2 U/mL of thrombin (Thr) pretreated with or without 100 nM PAR1 selective inhibitor (SCH79797, PAR1-i) for 1 h (n = 3). Representative image (upper panel) and quantification of gel areas (lower graph). The experiments were repeated three times. *, p < 0.05, and **, p < 0.01.

Article Snippet: On the day of experiments, cells in collagen gels were pretreated with the following reagents for 1 h: 100 nM PAR1 selective antagonist SCH79797 (No.1592 Tocris), 1 μM ROCK inhibitor Y-27632 (10005583, Cayman Chemical), 10 μM MLCK inhibitor, ML-7 (11801, Cayman Chemical), 10 μM of indomethacin (I-7378, Sigma), or 1 μM of progesterone (28921–64, Nacalai-tesque).

Techniques:

Journal: PLoS ONE

Article Title: Mechanisms of thrombin-Induced myometrial contractions: Potential targets of progesterone

doi: 10.1371/journal.pone.0231944

Figure Lengend Snippet: (A) Immunocytochemistry of PAR1 (red) and phosphorylated MLC2 (Ser19, green) in human myometrial cells. (B, C) Immunoblots of phosphorylated MLC2 (p-MLC2), total MLC2, and β-actin of myometrial cells. Myometrial cells were treated with thrombin (2 U/mL) as a function of time (B) , or pretreated with 100 nM PAR1 inhibitor (SCH79797) for 1 h, and then treated with 2 U/mL of thrombin for 30 min (C) . The experiments were repeated three times.

Article Snippet: On the day of experiments, cells in collagen gels were pretreated with the following reagents for 1 h: 100 nM PAR1 selective antagonist SCH79797 (No.1592 Tocris), 1 μM ROCK inhibitor Y-27632 (10005583, Cayman Chemical), 10 μM MLCK inhibitor, ML-7 (11801, Cayman Chemical), 10 μM of indomethacin (I-7378, Sigma), or 1 μM of progesterone (28921–64, Nacalai-tesque).

Techniques: Immunocytochemistry, Western Blot

Journal: PLoS ONE

Article Title: Mechanisms of thrombin-Induced myometrial contractions: Potential targets of progesterone

doi: 10.1371/journal.pone.0231944

Figure Lengend Snippet: (A) Collagen lattice assay of myometrial cells at 30 min with 2 U/mL of thrombin (Thr) pretreated with 1 μM progesterone (P4) for 1 h. Representative image (upper panel) and quantification of gel areas (lower graph). (B) Inhibition of thrombin-induced increases of PTGS2 , IL1B , and F2R mRNA by P4. Myometrial cells were pretreated with 1 μM of P4 for 1 h, and then treated with 2 U/mL of thrombin. (C) Gene expressions of progesterone receptor-A and–B ( PgR-A and PgR-B ) with 24 h treatment of 1 U/mL of thrombin, 10 nM of PGE2, and 10 nM of PGF2α (upper graphs) and PgR-A to PgR-B ratio (lower graphs). n = 3 in each group. *, p < 0.05, and **, p < 0.01. The experiments were repeated three times.

Article Snippet: On the day of experiments, cells in collagen gels were pretreated with the following reagents for 1 h: 100 nM PAR1 selective antagonist SCH79797 (No.1592 Tocris), 1 μM ROCK inhibitor Y-27632 (10005583, Cayman Chemical), 10 μM MLCK inhibitor, ML-7 (11801, Cayman Chemical), 10 μM of indomethacin (I-7378, Sigma), or 1 μM of progesterone (28921–64, Nacalai-tesque).

Techniques: Inhibition

Journal: Frontiers in Pharmacology

Article Title: TAK-442, a Direct Factor Xa Inhibitor, Inhibits Monocyte Chemoattractant Protein 1 Production in Endothelial Cells via Involvement of Protease-Activated Receptor 1

doi: 10.3389/fphar.2018.01431

Figure Lengend Snippet: FXa- and thrombin-induced MCP-1 production in HUVECs. MCP-1 concentration was determined in cell supernatant collected 20 h after the addition of the agonist, FXa (A) or thrombin (B) . In inhibition studies (C,D) , each inhibitor was added 1 h prior to the addition of the agonist and effects of TAK-442 (TAK), melagatran (Mel), and vorapaxar (Vor) on MCP-1 productions induced by 1 U/mL FXa (C) and 0.3 U/mL thrombin (D) were measured. Data are shown as mean ± SEM ( n = 3). ∗ P ≤ 0.025 compared with the control value of cells treated without FXa or thrombin (one-tailed Williams’ test) (A,B) . ∗ P ≤ 0.025 and † P ≤ 0.05 compared with the control value of cells treated with FXa or thrombin and no inhibitor (one-tailed Williams’ test and Student’s t -test, respectively, following ANOVA) (C,D) .

Article Snippet: The selective FXa inhibitor TAK-442, 1-(1-{(2S)-3-[(6-chloro–2-naphthyl) sulfonyl]-2-hydroxypropanoyl}piperidin-4-yl)tetra hydropyrimidin-2(1H)-one, the selective thrombin inhibitor melagatran, N-((1R)-2-{(2S)–2-[({4-[amino(imino)methyl]benzyl}amino)carbonyl]azetidin-1-yl}-1–cyclohexyl-2-oxoethyl) glycine, and the selective PAR1 antagonist vorapaxar, ethyl(9-{(E)–2-[5-(3-fluorophenyl)pyridin-2-yl]vinyl}-1-methyl-3-oxododecahydronaphtho[2,3-c]furan-6-yl)carbamate, were synthesized at Takeda Pharmaceutical Co., Ltd. (Osaka, Japan).

Techniques: Concentration Assay, Inhibition, Control, One-tailed Test

Journal: Frontiers in Pharmacology

Article Title: TAK-442, a Direct Factor Xa Inhibitor, Inhibits Monocyte Chemoattractant Protein 1 Production in Endothelial Cells via Involvement of Protease-Activated Receptor 1

doi: 10.3389/fphar.2018.01431

Figure Lengend Snippet: Effects of TAK-442, melagatran, and vorapaxar on the intracellular calcium ion concentration ([Ca 2+ ]i) induced by FXa, thrombin, and SFLLRN-NH2 in human PAR1-transfected Chinese hamster ovary (hPAR1/CHO-K1) cells. Calcium signal was recorded after the addition of FXa (0.03 U/mL) (A) , thrombin (0.003 U/mL) (B) , or PAR1 agonist peptide SFLLRN-NH2 (3 nM) (C) using FRIPR. Each inhibitor, TAK-442, melagatran, or vorapaxar, was pre-incubated with the cells for 10 min before the treatment with each agonist. Data are expressed as the percentage inhibition of calcium signal obtained after the addition of agonist in inhibitor-treated wells ( n = 4) compared with control wells (no inhibitor added). The drug concentration need to suppress the [Ca 2+ ]i by 50% (IC 50 ) was determined.

Article Snippet: The selective FXa inhibitor TAK-442, 1-(1-{(2S)-3-[(6-chloro–2-naphthyl) sulfonyl]-2-hydroxypropanoyl}piperidin-4-yl)tetra hydropyrimidin-2(1H)-one, the selective thrombin inhibitor melagatran, N-((1R)-2-{(2S)–2-[({4-[amino(imino)methyl]benzyl}amino)carbonyl]azetidin-1-yl}-1–cyclohexyl-2-oxoethyl) glycine, and the selective PAR1 antagonist vorapaxar, ethyl(9-{(E)–2-[5-(3-fluorophenyl)pyridin-2-yl]vinyl}-1-methyl-3-oxododecahydronaphtho[2,3-c]furan-6-yl)carbamate, were synthesized at Takeda Pharmaceutical Co., Ltd. (Osaka, Japan).

Techniques: Concentration Assay, Transfection, Incubation, Inhibition, Control