Brivanib inhibits liver fibrosis induced by thioacetamide.

<p>(A) Histological analysis of livers from placebo and brivanib-treated (25 and 50 mg/kg) groups at 4 weeks after initiation of thioacetamide (TAA). Pictures of Sirius red and Masson's trichrome staining are taken at 100×, magnification. (B) The number of bands of bridging fibrosis per high power field were counted in images obtained at 100× magnification after Sirius red staining. There was a significant decrease in the number of bands in both the 25 mg/kg and 50 mg/kg brivanib groups, compared to placebo. (C) The hepatic level of <i>collagen Iα1</i> mRNA was measured by real time PCR in placebo, 25 mg/kg and 50 mg/kg brivanib groups (n = 6 per group) at 4 weeks after the initiation of TAA. There was a substantial reduction in <i>collagen Iα1</i> mRNA at both brivanib dose levels. (D) Western immunoblotting for α-SMA showing decreased whole liver α-SMA after treatment with brivanib.</p

Brivanib decreases the transcription of growth factors and their receptors in carbon tetrachloride.

<p>Hepatic levels of growth factors and growth factor receptor mRNA were measured by real time PCR in sham and CCl₄ mice treated with no brivanib or with brivanib 25 mg/kg, 50 mg/kg, or 100 mg/kg. In all of the sham experiments, higher concentrations of brivanib decreased the mRNA levels of the growth factors and their receptors. (A) mRNA levels of <i>PDGFB</i> are not affected by brivanib treatment in CCl₄. (B) mRNA levels of <i>PDGFRB</i> decrease as the concentration of brivanib increases in CCl₄. (C) mRNA levels of <i>TGFB1</i> decrease as the concentration of brivanib increases in CCl₄. (D) mRNA levels of <i>TGFBR2</i> are lower in brivanib-treated CCl₄ mice compared to no brivanib. (E) mRNA levels of <i>FGF2</i> are not affected by brivanib treatment in CCl₄. (F) mRNA levels of <i>FGFR2</i> are lower in brivanib-treated CCl₄ mice compared to no brivanib. (G, H) mRNA levels of <i>VEGFA</i> and <i>VEGFR2</i> decrease as the concentration of brivanib increases in CCl₄ mice.</p

Brivanib decreases viability of PDGF-BB treated LX-2 cells.

<p>(A) The effect of increasing concentrations of brivanib on the viability of LX-2 HSCs cultured in DMEM/10% FBS was assessed using Cell Counting Kit-8 (CCK-8). Cells showed a brivanib dose-dependent decrease in viability, with half maximal inhibitory concentration (IC₅₀) of 16.98 µM (95% CI 13.95 µM–20.67 µM). (B) Cells stimulated with 5 ng/ml PDGF-BB in serum-free medium showed a similar trend albeit more pronounced decrease in viability compared to cells cultured in 10% FBS. The IC₅₀ of LX-2 cells cultured in serum-free media supplemented with PDGF-BB was 8.79 µM (95% CI 7.80 µM–9.99 µM).</p

PDGF, VEGF and FGF2 all induce proliferation of human LX-2 hepatic stellate cells.

<p>Relative proliferation of LX-2 cells as assessed by BrdU incorporation after addition of TGF-β1 (A); PDGF (B); VEGF (C); or FGF (D). The LX-2 HSCs were starved for 24 hours and then treated with the respective cytokine or growth factor. BrdU incorporation was measured 72 hours later. Data shown are representative of four samples per group and are presented as mean ± SEM. *, P<0.05 (normalized to BrdU incorporation in the absence of the growth factor).</p

Brivanib does not inhibit TGF-β1-induced α-SMA expression in human LX-2 hepatic stellate cells.

<p>(A) The expression of α-SMA in LX-2 HSCs after TGF-β1 treatment was assessed by Western immunoblotting. HSCs were incubated with 10% or 1% FBS for 24 hours, and TGF-β1 was added at different concentrations. The lysate was extracted 24 hours after addition of TGF-β1. Peak α-SMA expression was seen after treatment with 2 ng/ml of TGF-β1. (B) To determine the effect of brivanib on TGF-β1-induced activation of HSCs as assessed by α-SMA expression, LX-2 HSCs were partially serum-starved by incubation with 1% FBS for 24 hours. Brivanib was then added at increasing concentrations 2 hours before addition of 2 ng/ml TGF-β1. Cell lysates were prepared 24 hours after adding TGF-β1 and analyzed by Western immunoblotting.</p

Brivanib inhibits unstimulated and PDGF, VEGF, or FGF2-stimulated LX-2 cell proliferation.

<p>The effect of brivanib on cell proliferation of LX-2 HSCs without growth factor (A). The effect of brivanib on LX-2 cell proliferation induced by 50 ng/ml PDGF (B); 1 ng/ml VEGF (C); or 10 ng/ml FGF (D). LX-2 HSCs were starved for 24 hours, brivanib was added at the indicated concentrations, and 2 hours later, the respective growth factor was added. BrdU incorporation was measured at 72 hours after the administration of growth factor. Data shown are representative of four samples per treatment group and are presented as mean ± SEM. ∞,P<0.05 (vs. without growth factor and without brivanib) and *, P<0.05 (vs. with growth factor and without brivanib).</p

Brivanib inhibits liver fibrosis induced by bile duct ligation.

<p>(A) Histological findings at 14 days after BDL. The Sirius red and trichrome images are taken at 100×. (B) The number of bands of bridging fibrosis per high power field were counted in images obtained at 100× magnification after Sirius red staining. (C) Hepatic levels of <i>collagen Iα1</i> mRNA were measured by real time PCR in placebo, 25 mg/kg and 50 mg/kg brivanib groups at 14 days after BDL (n = 6 per group). (D) Western immunoblotting for α-SMA showing decreased whole liver α-SMA after treatment with brivanib. The expression of α-SMA in lysates extracted from liver tissue following sham ligation of the bile duct was measured by Western immunoblotting. β-actin is shown to control for loading. The sham controls confirm that brivanib does not induce liver fibrosis.</p

Brivanib inhibits PDGF-BB induced phosphorylation of PDGFRβ in human LX-2 hepatic stellate cells.

<p>(A) The expression of phosphorylated (P-PDGFRβ) and total PDGFRβ (T-PDGFRβ) in HSCs after PDGF-BB treatment was assessed by Western immunoblotting. HSCs were incubated in 10% FBS-supplemented DMEM for 24 hours, followed by starvation in serum-free medium for 24 hours. PDGF-BB was added at 5 or 10 ng/ml, followed by extraction of protein lysate 1 or 5 minutes after induction with PDGF-BB. Both time- and dose-dependent increase in phosphorylated PDGFRβ were seen, with minimal change in total PDGFRβ. β-actin was used as a loading control. (B) To determine the effect of brivanib on PDGF-BB induced phosphorylation of PDGFRβ, LX-2 cells were serum starved, followed by treatment with 5, 10 or 20 µM of brivanib. Two hours after brivanib treatment, 5 ng/ml PDGF-BB was added, and protein lysates prepared after 5 minutes of PDGF-BB exposure. All doses of brivanib tested inhibited PDGF-BB induced phosphorylation of PDGFRβ. (C) Ratio of phosphorylated PDGFRβ relative to total PDGFRβ. Prior to calculation of the P-PDGFRβ/T-PDGFRβ ratio, protein levels were first normalized to β-actin, and then to the P-PDGFRβ or T-PDGFRβ level in their respective control studies. Data represents the mean of four replicate studies ± SEM.</p

Brivanib stimulates the transcription of growth factors and their receptors in bile duct ligated mice.

<p>Hepatic levels of growth factors and growth factor receptor mRNA were measured by real time PCR in sham and BDL mice treated with no brivanib or with brivanib 25/kg or 50 mg/kg. In all of the sham experiments, higher concentrations of brivanib decreased the mRNA levels of the growth factors and their receptors. (A, B) BDL mice treated with brivanib show a dose-dependent increase in mRNA levels of <i>PDGFB</i> and <i>PDGFRB</i>. (C, D) mRNA levels of <i>TGFB1</i> increases as the concentration of brivanib increases in BDL mice; mRNA levels of <i>TGFBR2</i> are higher in BDL mice treated with 25 mg/kg and 50 mg/kg of brivanib, compared to the no brivanib group. (E, F) mRNA levels of <i>FGF2</i> and <i>FGFR2</i> are slightly higher with brivanib compared to no brivanib in BDL mice. (G, H) the mRNA levels of <i>VEGFA</i> and <i>VEGFR2</i> are not affected by brivanib treatment in BDL mice.</p

Efficacy and safety of tamsulosin 0.4 mg single pills for treatment of Asian patients with symptomatic benign prostatic hyperplasia with lower urinary tract symptoms: a randomized, double-blind, phase 3 trial

<p><b>Objective:</b> To verify the efficacy and safety of tamsulosin 0.4 mg and tamsulosin 0.2 mg compared with those of placebo in patients with lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH).</p> <p><b>Methods:</b> A total of 494 patients from multiple centers participated in this double-blind, randomized, phase 3 trial. Eligible patients were randomly assigned to the tamsulosin 0.4 mg group, tamsulosin 0.2 mg group or placebo group. The International Prostate Symptom Score (IPSS), maximum flow rate (<i>Qmax</i>), post-void residual (PVR) urine volume, blood pressure, heart rate and adverse events were compared among the three groups at 4, 8 and 12 weeks.</p> <p><b>Results:</b> A total of 494 BPH patients were analyzed. There were no differences in the baseline characteristics among the three groups. After 12 weeks of treatment, total IPSS was improved in the 0.2 mg and 0.4 mg tamsulosin groups; however, the extent of improvement was greater in the 0.4 mg group than in the 0.2 mg group (0.4 mg: −9.59 vs. 0.2 mg: −5.61; least-squares mean difference [95% confidence interval]: −3.95 [−5.01, −2.89], <i>p</i> < .0001). In addition, in the patients with severe symptoms (IPSS ≥20), total IPSS was improved the most in the 0.4 mg group (−11.27 ± 5.00, <i>p</i> < .0001). <i>Qmax</i> and PVR were improved in the 0.4 mg and 0.2 mg groups; however, the differences were not statistically significant between treatment groups. No patients experienced any serious adverse effects in any of the three groups.</p> <p><b>Conclusions:</b> Tamsulosin 0.4 mg and 0.2 mg appear to be superior to placebo treatment, and tamsulosin 0.4 mg is more effective than 0.2 mg in terms of total IPSS improvement. Tamsulosin 0.4 mg has favorable efficacy and tolerability in Asian men with symptomatic BPH.</p> <p><b>ClinicalTrials.gov Identifier:</b> NCT02390882.</p