1,729,931 research outputs found
Different routes of administering EW-7197 versus EW-7197·HBr for preventing peritoneal adhesion in a rat model
No full textBackground: The relatively low aqueous solubility of EW-7197 that was administered orally may have affected the desired concentration in the systemic circulation for treating peritoneal adhesion. This experimental study aimed to compare the efficacy of different routes of administering EW-7197 (2-fluoro-N-[(5-[6-methylpyridin-2-yl]-4-[(1,2,4)triazolo(1,5-a)pyridin-6-yl]-1H-imidazol-2-yl)methyl]aniline) and EW-7197·hydrobromide (HBr), with improved aqueous solubility, for inhibiting peritoneal adhesion in a rat model.
Methods: After peritoneal adhesion induction, 30 male Sprague-Dawley rats were randomly divided into 5 groups with 6 rats in each: group A, sham control; group B, orally administered 25 mg/kg of EW-7197·HBr for 7 days; group C, locally administered 25 mg/kg of EW-7197·HBr; group D, orally administered 20 mg/kg of EW-7197 for 7 days; and group E, locally administered 20 mg/kg of EW-7197. Gross examination, histologic staining (hematoxylin and eosin and Masson's trichrome), and immunohistochemical analyses (Ki-67 and α-smooth muscle actin marker [α-SMA]) were performed to evaluate the efficacy of both drugs.
Results: All procedures were technically successful. All treatment groups, except for group C, showed significantly reduced incidence, quality, tenacity, fibrosis, and collagen deposition scores and lowered expressions of Ki-67- and α-SMA-positive cells compared with group A. When comparing between groups, all scores were significantly lower in group B than in group C (all P .05).
Conclusion: Orally administering EW-7197·HBr and both orally and locally administering EW-7197 significantly prevented peritoneal adhesion formation, and orally administering EW-7197·HBr was the most effective overall
EW‐7197 prevents ulcerative colitis‐associated fibrosis and inflammation
No full textEW-7197 is a transforming growth factor-ß type I receptor kinase inhibitor with potential anti-inflammatory and antifibrotic properties. Here, we investigate the potential therapeutic effects of EW-7197 in a murine model of ulcerative colitis. EW-7197 attenuated the colitis disease activity index by improving rectal bleeding, body weight, and degree of stool consistency. EW-7197 also reduced colorectal tissue damage and the colon histopathological score by reducing crypt loss, mucosal damage, and tissue inflammation. Moreover, EW-7197 appeared to ameliorate the inflammatory and fibrotic responses by reducing oxidative stress, reducing submucosal edema and inflammatory cell infiltration, downregulating proinflammatory and pro-fibrotic genes, and inhibiting excessive collagen deposition in inflamed and fibrotic ulcerative colitis tissues. These results suggest that EW-7197 has potentially useful therapeutic properties against colitis, with clinically translational potential of inhibiting key pathological responses of inflammation and fibrosis in patients with colitis
EW-7197 prevents ulcerative colitis-associated fibrosis and inflammation
No full textEW-7197 is a transforming growth factor-ß type I receptor kinase inhibitor with potential anti-inflammatory and antifibrotic properties. Here, we investigate the potential therapeutic effects of EW-7197 in a murine model of ulcerative colitis. EW-7197 attenuated the colitis disease activity index by improving rectal bleeding, body weight, and degree of stool consistency. EW-7197 also reduced colorectal tissue damage and the colon histopathological score by reducing crypt loss, mucosal damage, and tissue inflammation. Moreover, EW-7197 appeared to ameliorate the inflammatory and fibrotic responses by reducing oxidative stress, reducing submucosal edema and inflammatory cell infiltration, downregulating proinflammatory and pro-fibrotic genes, and inhibiting excessive collagen deposition in inflamed and fibrotic ulcerative colitis tissues. These results suggest that EW-7197 has potentially useful therapeutic properties against colitis, with clinically translational potential of inhibiting key pathological responses of inflammation and fibrosis in patients with colitis
TGF-β type i receptor kinase inhibitor EW-7197 suppresses cholestatic liver fibrosis by inhibiting HIF1α-induced epithelial mesenchymal transition
No full textBackground/Aims: Hypoxia is an environmental factor that aggravates liver fibrosis. HIFla activates hepatic stellate cells (HSCs) and increases transforming growth factor-ß (TGF-β) signaling and the epithelial mesenchymal transition (EMT), accelerating the progression of fibrosis. We evaluated the anti-fibrotic therapeutic potential of a small-molecule inhibitor of TGF-β type I receptor kinase, EW-7197, on HIF1α-derived TGF-β signaling in cholestatic liver fibrosis. Methods: We used a bile duct ligation (BDL)-operated rat model to characterize the role of HIF1α-derived TGF-β signaling in liver fibrosis. Cellular assays were performed in LX-2 cells (human immortalized HSCs). The anti-fibrotic effects of EW-7197 in livertissues and HSCs were investigated via biochemical assays, immunohistochemistry (IHC), immunofiuorescence (IF), chromatin immunoprecipitation (ChIP) assays, real-time PCR, and western blotting. Results: In our BDL rat model, orally administered EW-7197 inhibited fibrosis and attenuated HIF1α-induced activation of HSCs and EMT in vivo. In addition, EW-7197 inhibited HIF1α-derived HSC activation and expression of EMT markers in LX-2 cells in vitro. Conclusion: This study suggests that EW-7197 exhibits potential as a treatment for liver fibrosis because it inhibits HIF1α-induced TGF-β signaling. © 2016 The Author(s)
EW-7197, a novel ALK-5 kinase inhibitor, potently inhibits breast to lung metastasis.
Get PDFAdvanced tumors produce an excessive amount of transforming growth factor β (TGFβ), which promotes tumor progression at late stages of malignancy. The purpose of this study was to develop anti-TGFβ therapeutics for cancer. We synthesized a novel small-molecule TGFβ receptor I kinase (activin receptor-like kinase 5) inhibitor termed N-[[4-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl]methyl]-2-fluoroaniline (EW-7197), and we investigated its potential antimetastatic efficacy in mouse mammary tumor virus (MMTV)/c-Neu mice and 4T1 orthotopic-grafted mice. EW-7197 inhibited Smad/TGFβ signaling, cell migration, invasion, and lung metastasis in MMTV/c-Neu mice and 4T1 orthotopic-grafted mice. EW-7197 also inhibited the epithelial-to-mesenchymal transition (EMT) in both TGFβ-treated breast cancer cells and 4T1 orthotopic-grafted mice. Furthermore, EW-7197 enhanced cytotoxic T lymphocyte activity in 4T1 orthotopic-grafted mice and increased the survival time of 4T1-Luc and 4T1 breast tumor-bearing mice. In summary, EW-7197 showed potent in vivo antimetastatic activity, indicating its potential for use as an anticancer therapy.ope
TGF-beta Type I Receptor Kinase Inhibitor EW-7197 Suppresses Cholestatic Liver Fibrosis by Inhibiting HIF1 alpha-Induced Epithelial Mesenchymal Transition
No full textBackground/Aims: Hypoxia is an environmental factor that aggravates liver fibrosis. HIF1 alpha activates hepatic stellate cells (HSCs) and increases transforming growth factor-beta (TGF-beta) signaling and the epithelial mesenchymal transition (EMT), accelerating the progression of fibrosis. We evaluated the anti-fibrotic therapeutic potential of a small-molecule inhibitor of TGF-beta type I receptor kinase, EW-7197, on HIF1 alpha-derived TGF-beta signaling in cholestatic liver fibrosis. Methods: We used a bile duct ligation (BDL)-operated rat model to characterize the role of HIF1 alpha-derived TGF-beta signaling in liver fibrosis. Cellular assays were performed in LX-2 cells (human immortalized HSCs). The anti-fibrotic effects of EW-7197 in liver tissues and HSCs were investigated via biochemical assays, immunohistochemistry (IHC), immunofluorescence (IF), chromatin immunoprecipitation (ChIP) assays, real-time PCR, and western blotting. Results: In our BDL rat model, orally administered EW-7197 inhibited fibrosis and attenuated HIF1 alpha-induced activation of HSCs and EMT in vivo. In addition, EW-7197 inhibited HIF1 alpha-derived HS[activation and expression of EMT markers in LX-2 cells in vitro. Conclusion: This study suggests that FW-7197 exhibits potential as a treatment for liver fibrosis because it inhibits HIF1 alpha-induced TGF-beta signaling. (C) 2016 The Author(s) Published by S. Karger AG, Base
Application of HPLC methods for determination of EW-7197 and its salt in rat plasma and comparison of their pharmacokinetics
No full text본 연구에서는 쥐의 혈장에서의 EW-7197을 검출하기 위하여 HPLC-UV 분석법을 확립 및 검증하였으며, EW-7197의 free form과 salt form을 정맥 및 경구로 투여하였을 때 EW-7197의 체내 동태에 어떠한 영향을 미치는지를 알아보고자 하였다. 또한, 생명공학연구원에서 얻은 같은 쥐의 혈장에서의 HPLC-MS/MS 결과를 토대로 본 연구에서 확립한 HPLC-UV 결과와 비교하여 좀더 간편하고 사용하기 쉬운 분석법을 개발하고자 하였다.
쥐의 혈장에서의 EW-7197을 정량하기 위하여 혈장 중 EW-7197과 내부표준물질인 EW-7214를 추출액인 메탄올을 사용하여 한번에 추출하였다. EW-7197과 내부표준 물질의 유지시간은 각각 19.4분 및 30.7분이었으며, 이 분석조건에서 EW-7197과 내부표준 물질은 기타 혈장성분들과 잘 분리되었다. EW-7197의 검량선은 0.05-10 μg/mL의 농도 범위에서 양호한 직선성을 나타내었으며, 정량한계를 포함한 농도 (0.05 μg/mL)에서 일내 및 일간 정밀성은 11% 및 9.9% 이하이었고, 정확성은 90.2-100.4%를 나타내어 본 분석법이 혈장 중 EW-7197을 검출하는데 충분한 감도, 정확성 및 정밀성을 갖고 있음을 보여주었다.
EW-7197과 EW-7197•HCl을 10 mg/kg로 경구 투여한 결과, EW-7197의 경우 최고 혈장 중 농도(Cmax)가 EW-7197•HCl에 비해 현저히 감소하였고 최고 혈장 중 농도에 도달하는 시간 (Tmax)은 현저히 증가하였으나, 그 외 모든 약동학적 파라미터들과 절대 생체이용률에서 있어서 EW-7197와 EW-7197•HCl 간에 유의한 차이는 나타나지 않았다.
EW-7197을 HPLC-UV로 정량하였을 때의 정량한계는 0.05 μg/mL이었고, HPLC-MS/MS로 정량하였을 때의 정량한계는 0.0195 μg/mL로 HPLC-MS/MS로 더 낮은 농도까지 정량이 가능하였지만, 두 분석법에서 얻은 EW-7197의 혈장 중 약물농도의 경시적 변화 및 약동학적 파라미터들에서 통계적으로 유의한 차이가 나타나지 않았다. 이를 통해서 HPLC-UV로도 충분히 정확하고 정밀한 EW-7197 분석이 가능함이 증명되었다.
결론적으로 본 연구에서 확립한 EW-7197의 HPLC-UV 분석법은 HPLC-MS/MS 방법보다 감도는 약간 떨어지나 특이성, 직선성, 정밀성 및 정확성이 검증되었고, 약동학 실험 후 얻어진 혈장 샘플에서 EW-7197 농도를 검출하는데 있어 HPLC-MS/MS 분석법과 큰 차이 없이 효과적이라는 것을 입증하였다. 또한 EW-7197은 물에 난용성이나 경구로 투여 시 위산에서 충분히 녹아 위장관 흡수가 가능하고 Cmax 및 Tmax를 제외한 다른 약동학적 파라미터들에 영향을 미치지 않으므로, 유기 용매 하에서 EW-7197를 salt form으로 만들기 위한 노력이 더 이상 필요하지 않다는 결론을 얻게 되었다.;The purpose of this study was to evaluate pharmacokinetic properties of EW-7197 with poor water solubility and EW-7197•HCl with improved water solubility. Despite improved solubility profiles in EW-7197•HCl, we investigated whether the oral administration of EW-7197 has advantages over the use of EW-7197•HCl in terms of its purity and its use in dosage forms. When EW-7197 was given orally to rats, its Tmax (0.44 ± 0.22 h) was significantly longer than that of EW-7197•HCl (0.18 ± 0.07 h; p < 0.05) and the Cmax of EW-7197 (0.81 ± 0.65 μg/mL) was significantly lower than that of EW-7197•HCl (1.79 ± 0.96 μg/mL; p < 0.05). Because EW-7197 needed dissolving time in gastrointestinal (GI) fluid, its Tmax was longer and the Cmax was lower than its salt form. However, there were no statistically significant differences between the two groups on the other pharmacokinetic parameters thereby significant difference was not observed in absolute bioavailability (%) between EW-7197 (34.8%) and EW-7197•HCl (43.0%).
In the present study, HPLC-UV method was used and validated for the determination of EW-7197 in rat plasma. In the sensitivity test, the LLOQ of HPLC-UV method was determined to be 0.05 μg/mL, whereas the LLOQ of HPLC–MS/MS method was determined to be 0.0195 μg/mL. HPLC-MS/MS method could determine lower LLOQ than HPLC-UV method but there were no statistically significant differences between two methods on the mean plasma concentration-time profiles and the all pharmacokinetic parameters. The close agreement of results obtained by the HPLC-UV and HPLC-MS/MS methods indicated that HPLC-UV method could provide excellent sensitivity and selectivity for the analysis of EW-7197 in rat plasma.
In conclusion, a simple HPLC-UV method was developed and validated successfully for the analysis of EW-7197 compared to HPLC-MS/MS method. Through the present study, we could develop validated HPLC-UV method and select EW-7197 to conduct the preclinical trial for developing a novel ALK5 inhibitor as a solid dosage form.I. INTRODUCTION 1
II. MATERIALS AND METHODS 8
1. Materials 8
2. Instruments 9
3. Animal Experiments 10
3.1 Preparation of rats 10
3.2 Sample formulation 11
3.3 Sample administration 11
4. HPLC Analysis 13
4.1 Preparation of stock and standard solutions 13
4.2 HPLC conditions 13
4.3 Sample preparation procedure 14
5. HPLC Validation 15
5.1 Specificity 15
5.2 Linearity 15
5.3 Precision and Accuracy 16
5.4 Sensitivity 16
5.5 Stability 16
6. Pharmacokinetic Analysis 18
7. Statistical Analysis 19
III. RESULTS 20
1. HPLC-UV Method Validation 20
1.1 Specificity 20
1.2 Linearity 22
1.3 Precision and Accuracy 23
1.4 Sensitivity 25
1.5 Stability 25
2. Pharmacokinetics of EW-7197 and EW-7197HCl 27
3. Comparison of Pharmacokinetic Results Analyzed by HPLC-UV and HPLC-MS/MS Methods 30
IV. DISCUSSION 37
REFERENCES 40
국문초록 4
Novel oral transforming growth factor- signaling inhibitor EW-7197 eradicates CML-initiating cells
No full textRecent strategies for treating CML patients have focused on investigating new combinations of tyrosine kinase inhibitors (TKIs) as well as identifying novel translational research agents that can eradicate CML leukemia-initiating cells (CML-LICs). However, little is known about the therapeutic benefits such CML-LIC targeting therapies might bring to CML patients. In this study, we investigated the therapeutic potential of EW-7197, an orally bioavailable transforming growth factor- signaling inhibitor which has recently been approved as an Investigational New Drug (NIH, USA), to suppress CML-LICs in vivo. Compared to TKI treatment alone, administration of TKI plus EW-7197 to CML-affected mice significantly delayed disease relapse and prolonged survival. Notably, combined treatment with EW-7197 plus TKI was effective in eliminating CML-LICs even if they expressed the TKI-resistant T315I mutant BCR-ABL1 oncogene. Collectively, these results indicate that EW-7197 may be a promising candidate for a new therapeutic that can greatly benefit CML patients by working in combination with TKIs to eradicate CML-LICs
EW-7197, an oral transforming growth factor beta type I receptor kinase inhibitor, for preventing peritoneal adhesion formation in a rat model
No full textBackground: EW-7197 is an oral transforming growth factor beta type I receptor kinase inhibitor currently undergoing phase I clinical trials for cancer treatment in the United States. This study evaluates whether EW-7197 prevents peritoneal adhesion formation in a rat model. Methods: Forty-eight female Wistar rats underwent peritoneal adhesion induction by the creation of peritoneal ischemic buttons and were randomly divided into 4 groups of 12 each. The control group received 0.3 mL vehicle by oral gavage once daily for 7 days after adhesion induction. The 10 mg and 20 mg groups received 10 or 20 mg/kg EW-7197 phosphate dissolved in 0.3 mL vehicle by oral gavage once daily for 7 days after adhesion induction. The rebound group received 20 mg/kg EW-7197 phosphate dissolved in 0.3 mL vehicle by oral gavage once daily for 7 days after adhesion induction followed by 0.3 mL vehicle only by gavage once daily for an additional 21 days. After the respective treatments were completed, the animals were euthanized. Results: All rats survived until the end of the study without complications. EW-7197 reduced the incidence, quality, and tenacity of peritoneal adhesions in a dose-dependent manner. Fibrosis and collagen production were reduced in EW-7197-treated peritoneal ischemic buttons. Transforming growth factor beta i/Smad2/3 signaling and mesothelial-to-mesenchymal transition were inhibited in EW-7197-treated peritoneal ischemic buttons. Discontinuation of EW-7197 was not associated with rebound effects. Conclusion: EW-7197 prevented peritoneal adhesion formation potentially via inhibition of transforming growth factor beta 1/Smad2/3-induced mesothelial-to-mesenchymal transition in a rat model. (C) 2018 Elsevier Inc. All rights reserved
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