Hsiang-Wen Tseng

Synthesis and structure-activity relationship of 6-arylureido-3-pyrrol-2-ylmethylideneindolin-2-one derivatives as potent receptor tyrosine kinase inhibitors.

A series of new ureidoindolin-2-one derivatives were synthesized and evaluated as inhibitors of receptor tyrosine kinases. Investigation of structure-activity relationships at positions 5, 6, and 7 of the oxindole skeleton led to the identification of 6-ureido-substituted 3-pyrrolemethylidene-2-oxindole derivatives that potently inhibited both the vascular endothelial growth factor receptor (VEGFR) and platelet-derived growth factor receptor (PDGFR) families of receptor tyrosine kinases. Several derivatives showed potency against the PDGFR inhibiting both its enzymatic and cellular functions in the single-digit nanomolar range. Among them, compound 35 was a potent inhibitor against tyrosine kinases, including VEGFR and PDGFR families, as well as Aurora kinases. Inhibitor 36 (non-substituted on the pyrrole or phenyl ring) had a moderate pharmacokinetic profile and completely inhibited tumor growth initiated with the myeloid leukemia cell line, MV4-11, in a subcutaneous xenograft model in BALB/c nude mice.

Development of novel 3D-QSAR combination approach for screening and optimizing B-Raf inhibitors in silico.

B-Raf is a member of the RAF family of serine/threonine kinases: it mediates cell division, differentiation, and apoptosis signals through the RAS-RAF-MAPK pathway. Thus, B-Raf is of keen interest in cancer therapy, such as melanoma. In this study, we propose the first combination approach to integrate the pharmacophore (PhModel), CoMFA, and CoMSIA models for B-Raf, and this approach could be used for screening and optimizing potential B-Raf inhibitors in silico. Ten PhModels were generated based on the HypoGen BEST algorithm with the flexible fit method and diverse inhibitor structures. Each PhModel was designated to the alignment rule and screening interface for CoMFA and CoMSIA models. Therefore, CoMFA and CoMSIA models could align and recognize diverse inhibitor structures. We used two quality validation methods to test the predication accuracy of these combination models. In the previously proposed combination approaches, they have a common factor in that the number of training set inhibitors is greater than that of testing set inhibitors. In our study, the 189 known diverse series B-Raf inhibitors, which are 7-fold the number of training set inhibitors, were used as a testing set in the partial least-squares validation. The best validation results were made by the CoMFA09 and CoMSIA09 models based on the Hypo09 alignment model. The predictive r(2)(pred) values of 0.56 and 0.56 were derived from the CoMFA09 and CoMSIA09 models, respectively. The CoMFA09 and CoMSIA09 models also had a satisfied predication accuracy of 77.78% and 80%, and the goodness of hit test score of 0.675 and 0.699, respectively. These results indicate that our combination approach could effectively identify diverse B-Raf inhibitors and predict the activity.

Antitumor Effect of Periplocin in TRAIL-Resistant Human Hepatocellular Carcinoma Cells through Downregulation of IAPs

Cortex periplocae is the dried root bark of Periploca sepium Bge., a traditional Chinese herb medicine. It contains high amounts of cardiac glycosides. Several cardiac glycosides have been reported to inhibit tumor growth or induce tumor cell apoptosis. We extracted and purified cortex periplocae and identified periplocin as the active ingredient that inhibited the growth of TNF-related apoptosis-inducing ligand-(TRAIL-) resistant hepatocellular carcinoma cells. The antitumor activity of periplocin was further increased by TRAIL cotreatment. Periplocin sensitized TRAIL-resistant HCC through the following two mechanisms. First, periplocin induced the expression of DR4 and FADD. Second, the cotreatment of TRAIL and periplocin suppressed several inhibitors of apoptosis (IAPs). Both mechanisms resulted in the activation of caspase 3, 8, and 9 and led to cell apoptosis. In addition, intraperitoneal injection (IP) of periplocin repressed the growth of hepatocellular carcinoma (HCC) in xenograft tumor model in mice. In summary, periplocin sensitized TRAIL-resistant HCC cells to TRAIL treatment and resulted in tumor cell apoptosis and the repression of tumor growth in vivo.

Bioisosteric replacement of an acylureido moiety attached to an indolin-2-one scaffold with a malonamido or a 2/4-pyridinoylamido moiety produces a selectively potent Aurora-B inhibitor.

Bioisosteric replacement of acylureido moiety in 6-acylureido-3-pyrrolylmethylidene-2-oxoindoline derivatives resulted in a series of malonamido derivatives with indolin-2-one scaffold (11-14). Further conformational restrictions of the malonamido moiety led to 2-oxo-1,2-dihydropyridine (21-25) or a 4-oxo-1,4-dihydropyridine derivatives (31-36). 4-Oxo-1,4-dihydropyridine derivatives were more potent Aurora B inhibitors than their 2-oxo-1,2-dihydropyridine counterparts and demonstrated cytotoxicities against A549 and HepG2 cells in the submicromolar range. In A549 cells, 31h decreased phosphorylation of histone H3, triggered polyploidy, induced expression of pro-apoptotic Fas and FasL with subsequent activation of caspase 8, resulting into apoptosis. In a Huh7-xenograft mouse model, 31h demonstrated potent in vivo efficacy with a daily dose of 5 mg/kg.

Novel azulene-based derivatives as potent multi-receptor tyrosine kinase inhibitors.

A series of azulene-based derivatives were synthesized as potent inhibitors for receptor tyrosine kinases such as FMS-like tyrosine kinase 3 (FLT-3). Systematic side chain modification of prototype 1a was carried out through SAR studies. Analogue 22 was identified from this series and found to be one of the most potent FLT-3 inhibitors, with good pharmaceutical properties, superior efficacy, and tolerability in a tumor xenograft model.

Abstract B186: ITRI-2531 - A novel kinase inhibitor for treating hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common cancers and is also a leading cause of cancer-related deaths worldwide. Until now, treatments for HCC remain limited, especially for the shortage of promising systemic therapies to replace systemic chemotherapy. Sorafenib, the first drug applied in targeted therapy for HCC, was approved and drew considerable attention. However, the efficacy and side effects of sorafenib are still unsatisfied, which make sorafenib far from ideal for treatment of HCC. Thus, developing novel therapeutics for HCC is necessary. A novel compound, ITRI-2531, is developed to improve the efficacy from sorafenib. ITRI-2531 shows cytotoxicity in HCC cell lines (Huh-7 and PLC/PRF/5) and patient-derived HCC tissue cell lines. In studies of kinase activities, ITRI-2531 suppresses the phosphorylation of MEK and ERK in PLC/PRF/5 cells and shows inhibition of multiple kinases (FLT3, FLT4, PDGFRA, PGDFRB, and VEGFR2) in KINOMEscanTM study. In in vivo studies, oral treatment of ITRI-2531 repressed subcutaneous Huh-7 and PLC/PRF/5 tumor growth in severe combined immunodeficient (SCID) mice and shows better efficacy than sorafinib. Moreover, ITRI-2531 prolongs the survival of SCID mice with orthotopic patient-derived xenograft tumor and suppresses alpha-fetoprotein in serum. On the other hand, ITRI-2531 also has good pharmacokinetic profiles. According to these results, we believe that ITRI-2531 warrants further evaluation as a new anti-HCC drug. Citation Format: Tsung-Keng Kuo, On Lee, Mai-Wei Lin, Li-Zong Lin, Chun-Min Liu, Tai-ju Hsieh, Chun-Chung Wang, Shyh-Horng Lin, Chia-Ni Chang, Hui-Chun Hsu, Nien-Tzu Chou, Chin-Pen Lai, Chih-Hung Chen, Chia-Mu Tu, Shih-Ta Chen, Yuan-Jang Tsai, Chih-Peng Liu, Jenn-Tsang Hwang, Jui-Wen Huang, Yen-Chun Chen, Chrong-Shiong Hwang, Hsiang-Wen Tseng. ITRI-2531 - A novel kinase inhibitor for treating hepatocellular carcinoma. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B186.

Abstract 5781: ITRI-260, a new azaazulenone class kinase inhibitor, induces complete tumor regression and prolongs survival in AML mice

The FMS-like tyrosine kinase-3 (FLT3) is a receptor tyrosine kinase with important roles in hematopoietic stem/progenitor cell survival and proliferation. FLT3 is over-expressed in the majority of acute myeloid leukemia (AML) and the presence of FLT3 active mutations is associated with poor prognosis, implicating FLT3 as a potential target for AML treatment. ITRI-260 is a new azaazulenone class kinase inhibitor that inhibited FLT3-ITD-dependent MV4-11 cell growth with IC50 of 21 nM and cellular phosphorylation with IC50 of 17 nM. In in vivo PK study, ITRI-260 showed favorable oral PK profiles. The maximum concentration of ITRI-260 was 700 nM after a single 100 mg/kg oral dosing in mice, with dose proportional exposure from 10 to 150 mg/kg. ITRI-260 showed therapeutic efficacy in a human AML orthotopic SCID mouse model, following a 100 mg/kg/qd oral dosing regimen. The treatment group showed 100% survival of animals at day 90 after tumor cell inoculation, while the vehicle control group showed 100% mortality. In the other study, ITRI-260 also increased survival in primary tumor cell engraftment model inoculated with cells isolated from sunitinib- and sorafenib-resistant AML patient. Through hERG assay with patch clamp technology, we found higher IC50 of ITIR-260 compared with sunitinib and MLN-518, which indicates that ITRI-260 might arise less cardiotoxic concern. In a 14-day subchronic toxicology study in mice, ITRI-260 was generally tolerated at 300 mg/kg/qd. The most prominent adverse effect appeared at 600 mg/kg/qd group is starry sky appearance in spleen, which is moderate and reversible. No specific clinical signs or body weight loss were attributed to the test article. In conclusion, the in vitro/in vivo PK/PD/toxicology studies have provided the basis for the clinical developmental potential of ITRI-260 in AML. The bridge studies from discovery to development such as CMC, GLP toxicology/safety pharmacology and pre-formulation are now in progress. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5781.

Abstract 3356: A controversial study on well reported liver cancer stem cell marker

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Recent studies suggest that the presence of cancer stem cells (CSC) could be linked with patients’ survival. The ability of cancers to grow indefinitely has fueled the idea that cancer and stem cells may have common underlying mechanisms. It has been suggested that tumors are initiated from cancer stem cells (CSCs) with proliferation potential drives the growth of cancer. CSCs are resistant to chemotherapy and radiation. However, the suggested cancer stem cell markers in human hepatocellular carcinoma cells are variable and confused. In this study, we profiled some of the most reported CSC markers, including CD133, epithelial cell adhesion molecule (EpCAM), aldehyde dehydrogenase (ALDH), CD90, CD24, c-kit, global-H and stemness genes in eight human hepatocellular carcinoma cell lines. Through specific marker expressed cell sorting by FACs aria or magnetic beads, the CSC associated drug resistance and tumorigenicity were further evaluated. However, there is no obvious difference among parental group, marker-positive group and marker-negative group in these CSC characteristics evaluated. It seems no good correlation between reported markers in liver cancer stem cells. Therefore, presence of markers alone should be taken with caution as single prognostic parameters. Through harsh culture condition, spheroid cell grew and had been isolated, which perform CSC-like properties. Moreover, forced activation of an ESC-like gene expression program can reprogram HCC cells into CSC-like cells and achieve pathologic self-renewal. The ability to create induced cancer stem cells (iCSC) may provide opportunities to better define the biology of cancer stem cells in order to trace or eliminate them in human patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3356.