bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1698: Identification of drugs with eribulin combinatorial activity that kill both eribulin-sensitive and eribulin-insensitive tumor cells
Authors: Uenaka Toshimitsu , Rickles Richard , Funahashi Yasuhiro , Zhu Ping , Grenier Jill M. , Steiger Janine , Zhao Nanding , Littlefield Bruce A. , Matsui Junji , Nomoto Kenichi
Published: May 2015

Eribulin mesylate, a nontaxane microtubule dynamics inhibitor with a mechanism distinct from most other antitubulin therapeutics, is clinically used in the United States for treatment of certain patients with locally advanced or metastatic breast cancer. Eribulin has broad anticancer activity in a wide variety of preclinical cancer models; as a result, numerous clinical trials are ongoing to investigate efficacy in other non-breast tumor types. We have examined eribulin sensitivity in a panel of 25 human cancer cell lines representing a variety of tumor types albeit with a preponderance of breast and lung cancer cell lines. As expected, the cell lines vary in sensitivity to eribulin at clinically relevant concentrations. To identify combination drugs capable of increasing anticancer effects in patients already responsive to eribulin, as well as inducing de novo anticancer effects in non-responders, we performed a combinatorial high throughput screen to identify drugs that combine with eribulin to selectively kill tumor cells. Among other observations, we found that inhibitors of ErbB1/ErbB2 (lapatinib, BIBW-2992, erlotinib), MEK (E6201, trametinib), PI3K (BKM-120), mTOR (AZD 8055, everolimus), and PI3K/mTOR (BEZ 235), and a BCL2 family antagonist (ABT-263) show combinatorial activity with eribulin. In addition, antagonistic pairings with other agents, such as topoisomerase I inhibitors (topotecan hydrochloride), an HSP-90 inhibitor (17-DMAG), and gemcitabine and cytarabine, were also identified. In summary, the preclinical studies described here have identified several combination drugs that have the potential to either augment or antagonize eribulin's anticancer activity. Further elucidation of the mechanisms responsible for such interactions may be important for identifying valuable therapeutic partners for eribulin.

Citation Format: Toshimitsu Uenaka, Richard Rickles, Yasuhiro Funahashi, Ping Zhu, Jill M. Grenier, Janine Steiger, Nanding Zhao, Bruce A. Littlefield, Junji Matsui, Kenichi Nomoto. Identification of drugs with eribulin combinatorial activity that kill both eribulin-sensitive and eribulin-insensitive tumor cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1698. doi:10.1158/1538-7445.AM2014-1698
Abstract 17: Hepatitis C virus (HCV) core protein potentiates proangiogenic activity of hepatocellular carcinoma (HCC) cells
Authors: Shao Yu-Yun , Huang Chung-Yi , Chen Jun-Wei , Hsu Chih-Hung , Cheng Ann-Lii
Published: May 2015

Background: Recent clinical trials found that sorafenib may have better efficacy for HCV-related HCC than HCC with other etiologies. HCV core protein has been found to influence the mitogen-activated protein kinase pathway, which is one of the targets of sorafenib. We thus planned this study to examine the effect of HCV core protein overexpression on HCC cells.

Methods: We stably expressed HCV core protein in Huh7, a HCC cell line, using a lentivirus based vector S2. Two stable clones were selected for experiments. One clone (Huh7-core-high) expressed a higher amount of HCV core protein than the other (Huh7-core-low). Huh7 expressing the empty vector (Huh7-S2) served as a control. We compared the cytotoxicity of sorafenib in these cells using the MTT assay. Tube formation analysis and Matrigel plug assay were performed to evaluate the angiogenic activity.

Results: Huh7-core-high and Huh7-core-low cells did not show increased cell proliferation compared to Huh7-S2 cells. Sorafenib showed similar in vitro cytotoxicity for Huh7-core-high, Huh7-core-low, and Huh7-S2 cells. Overexpression of HCV core protein potentiates proangiogenic activity of Huh7 cells dose-dependently. In tube formation analysis, Huh7-core-high induced more potent angiogenesis than Huh7-S2 cells with more total tube lengths (p = 0.02), mean tube lengths (p = 0.04), mean tube areas (p = 0.04), branch points (p = 0.03), and nodes (p < 0.01). Huh7-core-low cells induced stronger angiogenesis than Huh7-S2 cells with more total tube lengths (p = 0.05) and branch points (p = 0.01). By Matrigel plug assay, Huh7-core-high and Huh7-core-low cells dose-dependently induced stronger angiogenesis in mice compared to Huh7-S2 cells with significantly higher hemoglobin levels in the Matrigel plugs.

Conclusions: HCV core protein expression potentiates the proangiogenic activity of HCC, both in vitro and in vivo. (This work was supported by grants NSC98-3112-B-002-038, NSC101-2314-B-002-141, 100CAP1020-2 & 102-2314-B-002-120.)

Citation Format: Yu-Yun Shao, Chung-Yi Huang, Jun-Wei Chen, Chih-Hung Hsu, Ann-Lii Cheng. Hepatitis C virus (HCV) core protein potentiates proangiogenic activity of hepatocellular carcinoma (HCC) cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 17. doi:10.1158/1538-7445.AM2014-17
Abstract 1700: A combination of two tropical spice compounds potentiates chemotherapy response in castration-resistant prostate cancer cells
Authors: Mendha Taaha A. , Nagarajarao Shamaladevi , Lokeshwar Balakrisha L.
Published: May 2015

Introduction: Patients with castration resistant advanced prostate cancer (CRPC) respond poorly to potent chemotherapeutic drugs (e.g., Docetaxel or Mitoxantrone) with survival advantages that last less than 4 months. Since chemotherapy itself has morbidity in the elderly patients, adding another such drug is unlikely to improve the outcome. We investigated the potential therapeutic benefits of treating CRPC with a combination of two natural dietary anti-proliferative compounds: Curcumin and Piperine. They are derived from the tropical spices, turmeric and black pepper, respectively. Both are orally bioavailable and can be consumed over a long period of time.

Materials and methods: The cytotoxicity of Curcumin, Piperine, and Docetaxel were individually tested on a CRPC cell line PC-3, which was incubated for 48 hours or 72 hours. Cytotoxicity was evaluated by cell counting, a colorimetric assay and by clonogenic survival, colony-formation assay. Enhancement of Docetaxel cytotoxicity by natural compounds was evaluated by treating cells with these compounds 24 hours before adding Docetaxel or with co-incubation. The potential mechanism of cytotoxicity was examined by an apoptosis assay and changes in cell cycle phase-fractions. The efficacy of the combination was evaluated by Isobologram analysis and ANOVA.

Results: All three drugs tested on PC-3 showed cytotoxicity, Docetaxel was the most cytotoxic (IC50∼ 25 nM) and Piperine was the least cytotoxic (IC50∼ 100 μM). Curcumin alone was significantly cytotoxic to PC-3 cells at 10 μM but it enhanced the cytotoxicity of Docetaxel. Piperine, when combined with Curcumin or Docetaxel, significantly increased the cytotoxicity by 50%. Prior exposure to Piperine resulted in enhanced cytotoxicity of Docetaxel and Curcumin more than when in co-incubation. The increased cytotoxicity in combination therapy was mainly due to increased apoptotic activity.

Conclusion: This study demonstrated that combining diet-derived antiproliferatinve compounds with a chemotherapy drug can potentially enhance tumoricidal activity and therefore, potentially reduce morbidity and mortality of patients with CRPC. Pretreatment with dietary compounds could potentially enhance subsequent treatment with chemotherapy agents.

Citation Format: Taaha A. Mendha, Shamaladevi Nagarajarao, Balakrisha L. Lokeshwar. A combination of two tropical spice compounds potentiates chemotherapy response in castration-resistant prostate cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1700. doi:10.1158/1538-7445.AM2014-1700
Abstract 1701: A synthetic flavonoid abrogates doxorubicin resistance through inhibition of focal adhesion kinase and P-glycoprotein activity in breast cancer cells
Authors: Datta Amrita , Rider Barbara J. , Elmageed Zakaria Y. Abd , Kim Hogyoung , Mondal Debasis , Mageed Asim B. Abdel
Published: May 2015

Chemotherapy is a mainstay in the treatment of breast cancer. FDA approved, Doxorubicin (Dox) is considered to be the most effective chemotherapeutic agent for the treatment of solid tumors like breast cancer. However, resistance to Dox impairs successful treatment. The increased expression of multidrug-resistance P-glycoprotein (P-gp) pump that constantly efflux drugs, such as Dox, is a well documented mechanism of resistance.

Flavonoids as a class have been implicated in the inhibition of ABC transporters such as BCRP, P-gp and MRP-1, have potent anxiolytic activity, protect against anthracycline induced cardiotoxicity, and improve general health.

Our current study proposes to examine the efficacy of a synthetic flavonoid, SF as a chemotherapeutic agent alone and in combination with Dox in a xenograft nude mouse model and to delineate the specific molecular targets involved. Treatment with a combination of Dox and SF lead to significant cell death in the Dox resistant, MCF-7/Dox cells as compared to treatment with Dox alone. In vivo studies using a xenograft nude mouse model corroborated these findings. Also, mice treated with SF alone and in combination with Dox showed improved overall health. SF also inhibited P-gp pump activity and increased Dox sequestration in cancer cells, leading to chemosensitization. Moreover, SF alone or in combination with Dox caused remarkable de-phosphorylation of constitutively active FAK(Focal Adhesion Kinase) in MCF-7/Dox cells. In conclusion, our study beings forth an excellent prototype candidate, SF- as an anti-tumor agent with little to no toxicity and provides new insights into previously uncharacterized mechanisms for the anti-cancer activity of SF alone and in combination with Dox in resistant breast cancer cells. The desirable traits of endowed pleiotropic activities, safety, synergistic action with therapeutic agents make SF attractive for clinical use.

Citation Format: Amrita Datta, Barbara J. Rider, Zakaria Y. Abd Elmageed, Hogyoung Kim, Debasis Mondal, Asim B. Abdel Mageed. A synthetic flavonoid abrogates doxorubicin resistance through inhibition of focal adhesion kinase and P-glycoprotein activity in breast cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1701. doi:10.1158/1538-7445.AM2014-1701
Abstract 1703: Nicotine-induced gemcitabine resistance is reversed by gamma-aminobutyric acid but enhanced by baclofen in pancreatic cancer xenografts and in pancreatic cancer cells in vitro
Authors: Banerjee Jheelam , Al-Wadei Hussein A. N , Al-Wadei Mohammed H. , Dagnon Koami , Schuller Hildegard M.
Published: May 2015

Pancreatic cancer is frequently resistant to cancer therapeutics. Smoking and alcoholism are risk factors and pancreatic cancer patients often undergo nicotine replacement therapy (NRT) and treatment for alcohol dependence. Based on our report that low dose nicotine within the range of NRT causes gemcitabine resistance in pancreatic cancer, our current study has tested the hypothesis that GABA or the selective GABA-B-R agonist baclofen used to treat alcohol dependence reverse nicotine-induced gemcitabine resistance in pancreatic cancer. Using pancreatic cancer cell lines BXPC-3 and PANC-1, our data show that GABA significantly reversed gemcitabine resistance induced by low dose nicotine in xenografts whereas baclofen did not. This effect of GABA was accompanied by decreases in cAMP, p-CREB, p-AKT, p-Src, p-ERK metalloproteinases-9 and -2 and EGR-1 and increases in cleaved caspase-3 in xenografts whereas baclofen had the opposite effects. In vitro exposure of cells to single doses or seven days of nicotine induced the protein expression of MMP-2, MMP-9 and EGR-1 and these responses were blocked by GABA. Baclofen downregulated the protein expression of GABA-B-Rs in xenograft tissues and in cells exposed to baclofen for seven days in vitro. This response was accompanied by inversed baclofen effects from inhibition of cAMP formation after single dose exposures to stimulation of cAMP formation in cells pretreated for seven days. These findings suggest GABA as a promising agent to overcome nicotine-induced gemcitabine resistance in pancreatic cancer whereas treatment of alcoholism by baclofen may increase gemcitabine resistance.

Supported by grants RO1CA130888 and RO1CA042829 with the National Cancer Institute.

Citation Format: Jheelam Banerjee, Hussein A. N Al-Wadei, Mohammed H. Al-Wadei, Koami Dagnon, Hildegard M. Schuller. Nicotine-induced gemcitabine resistance is reversed by gamma-aminobutyric acid but enhanced by baclofen in pancreatic cancer xenografts and in pancreatic cancer cells in vitro. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1703. doi:10.1158/1538-7445.AM2014-1703
Abstract 1704: Combination of Pirfenidone and Cisplatin as a potential therapeutic strategy in NSCLC
Authors: Mediavilla-Varela Melanie , Noyes David , Boateng Kingsley , Antonia Scott
Published: May 2015

Lung cancer is the leading cause of cancer death in the United States. Due to limited efficacy of current conventional chemotherapy, the majority of patients face poor prognosis and eventually succumb the disease. Therefore, the study of combination therapy using traditional anticancer drugs with novel therapeutic approaches is needed. Pirfenidone is a small molecule that is being used in Phase II and III clinical trials for idiopathic pulmonary fibrosis (IPF). Although it has already been approved for IPF in Europe, there is no known mechanism of action known for the drug. Recently we discovered that Pirfenidone has a detrimental effect on cancer associated fibroblasts (CAFs) as well as in non-small cell lung cancer (NSCLC) cells. Our hypothesis is that Pirfenidone, in combination with standard chemotherapy, will act synergistically and proffer a more potent therapeutic strategy for NSCLC. Our studies show that in a co-culture model there is a decrease in viability on CAFs and NSCLC cells when the combination of Pirfenidone and Cisplatin is used. Furthermore, Pirfenidone and Cisplatin were able to have an anti-tumor effect in a xenograft model of NSCLC and CAFs. In addition, insights in the mechanism by which Pirfenidone and Cisplatin induce synergy in NSCLC cells are being elucidated. Our studies show that the combination of drugs leads to an increase in phosphorylated ERK as well as a decrease in phosphorylated Akt. Thus, involving the PI3K and MAPK/ERK pathways in Pirfenidone/Cisplatin induced cell death. Our lab is the first to recognize the therapeutic potential of combining Pirfenidone and Cisplatin, as well as elucidating the mechanism of action for both drugs in NSCLC.

Citation Format: Melanie Mediavilla-Varela, David Noyes, Kingsley Boateng, Scott Antonia. Combination of Pirfenidone and Cisplatin as a potential therapeutic strategy in NSCLC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1704. doi:10.1158/1538-7445.AM2014-1704
Abstract 1708: Heparanase regulates response to chemotherapy in myeloma
Authors: Ramani Vishnu Prakash C. , Zhan Fenghuang , Tricot Guido , Sanderson Ralph D.
Published: May 2015

Following therapy, essentially all multiple myeloma patients relapse due to a population of chemoresistant cells leading to eventual patient death. Our lab has identified heparanase, an endoglycosidase, as a master regulator of aggressive myeloma disease. In the present study that was designed to probe the role of heparanase in chemoresistance, we made the striking finding that heparanase promotes survival of tumor cells post-therapy. Using an array of cellular viability assays, we tested for differences in viability and apoptosis between myeloma cells engineered to express high levels of heparanase (HPSE-high) and cells expressing low levels of heparanase (HPSE-low) following their exposure to frontline anti-myeloma drugs (bortezomib, carfilzomib). Results revealed a much smaller apoptotic fraction and higher cell viability following treatment of the HPSE-high cells compared to HPSE-low cells. In addition to promoting tumor survival, we also discovered that myeloma cell expression and secretion of heparanase is dramatically enhanced in response to chemotherapy. Using small molecule inhibitors, we were able to pinpoint that activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) was driving the heparanase expression upon chemotherapy. The secreted heparanase generated by chemotherapy was readily taken up by tumor cells growing in culture, consistent with previous studies demonstrating that heparanase is taken up by myeloma tumor cells and that this activates aggressive tumor behavior. Extending our findings to the clinic revealed that in tumor cells from seven out of nine myeloma patients examined, there was a dramatic increase in heparanase gene expression following chemotherapy. Together, these results demonstrate a pivotal role for heparanase in regulating myeloma response to therapy and indicate that the use of heparanase inhibitors may inhibit or delay relapse in myeloma.

Citation Format: Vishnu Prakash C. Ramani, Fenghuang Zhan, Guido Tricot, Ralph D. Sanderson. Heparanase regulates response to chemotherapy in myeloma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1708. doi:10.1158/1538-7445.AM2014-1708
Abstract 1727: Development of novel TAK1 inhibitors for pancreatic cancer
Authors: Chiao Paul J. , Zhuang Zhuonan , Xia Qianghua , Schuber Paul T. , Sun Duoli , Peng Zhenghong , Maxwell David S. , Bornmann William G.
Published: May 2015

Transforming growth factor- (TGF-) activation kinase (TAK1, MAP3K7) is a member of MAPKKK family [1]. It was well established that TAK1 plays a key role in both TNFα and IL-1 pathway. Knock down of TAK1 impaired the activation of IKK and JNK by TNFα and IL-1 [2, 3]. NF-B activation induced by TNFα and IL-1 are severely impaired in TAK1 knock out MEF cells [4]. TAK1 acts as the kinase of IKK and therefore regulates NF-κB activation. Our previous work has demonstrated that TAK1 is upregulated and plays an important role in chemotherapeutic drug resistance in pancreatic cancer cells. Block TAK1 activity can reverse pancreatic cancer cell resistance to chemotherapeutic drug and reduce tumor size in Orthotopic Xenograft models [5]. These findings underscore the significance of TAK1 in mediating drug resistance. Strategies to target TAK1 may play a critical role in the treatment of this deadly malignancy.

Utilizing computational-based structural modeling and a recently published crystal structure of TAK1, compounds were designed to inhibit the ATP-binding pocket of the TAK1. Synthetic templates were proposed for building a urea based library of potential inhibitors. A multiple stage docking involving both pre- and post-filtering and progressively more stringent docking criteria led to approximately 1,000 structures. The resulting scores from two docking programs were fed into workflow software that combined the results and calculated ADME properties. To provide better consistency in scoring and less bias in setting the filtering cutoffs, we also included docking decoys and utilized a more statistically relevant Z-score. After application of a final binding mode filter, there were 22 structures that were selected and ranked. A seven-step synthesis was developed and a focused library of compounds was synthesized from those top-ranked compounds.

Note: This abstract was not presented at the meeting.

Citation Format: Paul J. Chiao, Zhuonan Zhuang, Qianghua Xia, Paul T. Schuber, Duoli Sun, Zhenghong Peng, David S. Maxwell, William G. Bornmann. Development of novel TAK1 inhibitors for pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1727. doi:10.1158/1538-7445.AM2014-1727
Abstract 1739: Preclinical profile of BAY 1163877 - a selective pan-FGFR inhibitor in phase 1 clinical trial
Authors: Heroult Melanie , Ellinghaus Peter , Sieg Christian , Brohm Dirk , Gruenewald Sylvia , Collin Marie-Pierre , Boemer Ulf , Lobell Mario , Huebsch Walter , Ocker Matthias , Ince Stuart , Haegebarth Andrea , Jautelat Rolf , Hess-Stumpp Holger , Brands Michael , Ziegelbauer Karl
Published: May 2015

Fibroblast growth factors (FGFs) orchestrate a variety of cellular functions by binding to their transmembrane tyrosine-kinase receptors (FGFR1-4) and activating downstream signaling pathways. FGF signaling has been demonstrated to be altered in a high proportion of cancers, with activating mutations and/or overexpression of FGFRs frequently observed in lung, gastric, breast and urothelial tumors. Therefore, targeting FGFRs using selective FGFR inhibitors is an attractive therapeutic approach to treat cancer patients.

BAY 1163877 is as an orally available, selective and potent inhibitor of FGFR-1, -2 and -3 kinase activity. BAY 1163877 has been advanced through preclinical development and we disclose here the first details of its preclinical profile.

BAY 1163877 inhibited FGFR-1, -2, -3 kinase activity in the nanomolar range and demonstrated a kinase selectivity profile for FGFR-1, -2 and -3 over 222 kinases tested. BAY 1163877 inhibited proliferation of various cancer cell lines in vitro and phosphorylation of downstream signaling molecules. BAY 1163877 was also tested in vivo in monotherapy and combination therapy on various human xenografts and syngeneic tumors and inhibited growth of tumors presenting at least one FGFR alteration.Overall, the in vitro and in vivo studies confirm that the FGFR inhibitor BAY 1163877 is a potent and selective inhibitor of altered FGFRs pathways in cancer models. A Phase 1 clinical trial (NCT01976741) has been initiated.

Citation Format: Melanie Heroult, Peter Ellinghaus, Christian Sieg, Dirk Brohm, Sylvia Gruenewald, Marie-Pierre Collin, Ulf Boemer, Mario Lobell, Walter Huebsch, Matthias Ocker, Stuart Ince, Andrea Haegebarth, Rolf Jautelat, Holger Hess-Stumpp, Michael Brands, Karl Ziegelbauer. Preclinical profile of BAY 1163877 - a selective pan-FGFR inhibitor in phase 1 clinical trial. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1739. doi:10.1158/1538-7445.AM2014-1739
Abstract 1746: Novel and selective Axl inhibitors
Authors: Zhang Zaihui , Li Rick , Lee Erica , Hu Yuxiang , Yan Jun , Sanghera Jasbinder
Published: May 2015

Axl is a member of the TAM tyrosine kinase family and is activated by products of Gas6 and Protein S genes. Axl is overexpressed or overactive in breast, renal, endometrial, ovarian, thyroid, non-small cell lung carcinoma, uveal melanoma as well as in myeloid leukemias. It has also been demonstrated that Axl expression is upregulated in drug-resistant cancer cells. Knock-down of Axl by RNAi repressed tumor formation in MDA-231 breast carcinoma model. Downregulation of Axl expression by RNAi led to a decrease of K562/ADR and MCF-7/ADR cells invasion, proliferation, and increased cell chemosensitivity in vitro. In a xenograft model of MDR cells, downregulation of Axl enhanced the anticancer activity of chemotherapeutic drugs. Therefore, Axl is a potential target for the development of chemotherapeutic agents for cancer therapy. During our endeavor of discovering and developing Axl kinase inhibitors for cancer therapy, we have discovered a series of potent and selective Axl inhibitors. These new compounds have demonstrated nM potency against Axl and good selectivity against other kinases in a selected panel of kinases. Many of these new compounds can reduce the pAkt levels in several cancer cell lines in a dose dependent manner and demonstrated inhibition of cell proliferation at sub-micromolar concentration in thymidine incorporation assay. Several selected compounds have demonstrated efficacy in colony formation assays. The most promising compounds are now being selected for in vivo proof-of-concept studies. The pharmaceutical properties of these compounds together with ADME profile were also evaluated and will be reported. These new Axl inhibitors represent a new approach for cancer therapy.

Citation Format: Zaihui Zhang, Rick Li, Erica Lee, Yuxiang Hu, Jun Yan, Jasbinder Sanghera. Novel and selective Axl inhibitors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1746. doi:10.1158/1538-7445.AM2014-1746
Abstract 1750: Combined blockade of Aurora A and JAK2 kinase is highly effective at inhibiting malignant transformation
Authors: Sebti Said M. , Yang Hua , Lawrence Harshani , Kazi Aslamuzzaman , Gervaria Harsukh , Patel Ronil , Luo Yunting , Rix Uwe , Schonbrunn Ernst , Lawrence Nicholas
Published: May 2015

Our ongoing efforts to validate kinases for cancer therapy led to the novel finding that ectopic expression of JAK2 and Aurora A together is more effective than each alone at inducing normal cells to grow in an anchorage-independent manner and to invade. In addition, siRNA silencing or pharmacological inhibition of JAK2 and Aurora A with Ruxolitinib and Alisertib, respectively, is more effective than blocking each kinase alone at suppressing anchorage-dependent and -independent growth and invasion as well as at inducing apoptosis. This led us to develop dual Aurora and JAK inhibitors, AJI-214 and AJI-100, which potently inhibit the activities of Aurora A, Aurora B and JAK2 in vitro. In human cancer cells, these dual inhibitors block the auto-phosphorylation of Aurora A (Thr-288) and the phosphorylation of the Aurora B substrate histone H3 (Ser-10) and the JAK2 substrate STAT3 (Tyr-705). Furthermore, AJI-214 and AJI-100 inhibit anchorage dependent and independent cell growth and invasion and induce G2/M cell cycle accumulation and apoptosis. Finally, the more soluble AJI-100 was effective at inducing tumor regression of human xenografts in mice. Taken together, our genetic and pharmacological studies indicate that targeting Aurora A and JAK2 together is a more effective approach than each alone at inhibiting malignant transformation and warrant further advanced pre clinical investigations of dual Aurora A/JAK2 inhibitors as potential anti tumor agents.

Citation Format: Said M. Sebti, Hua Yang, Harshani Lawrence, Aslamuzzaman Kazi, Harsukh Gervaria, Ronil Patel, Yunting Luo, Uwe Rix, Ernst Schonbrunn, Nicholas Lawrence. Combined blockade of Aurora A and JAK2 kinase is highly effective at inhibiting malignant transformation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1750. doi:10.1158/1538-7445.AM2014-1750
Abstract 1755: CT053PTSA, a novel c-MET and VEGFR2 inhibitor, potently suppresses angiogenesis and tumor growth
Authors: Xi Ning , Zhang Yingjun , Wang Zhaohe , Wu Yanjun , Wang Tingjin
Published: May 2015

Background: In many human cancers, c-Met is activated via receptor overexpression, amplification, mutation and/or a ligand-dependent autocrine/paracrine loop. c-MET-driven tumor growth and metastasis may be further stimulated by hypoxic conditions in the tumor microenvironment, which are often exacerbated by the VEGF-pathway inhibition. Therefore, blockades of c-Met and VEGFR2 simultaneously may yield synergistic inhibition on tumor growth.

Methods and Results: CT053PTSA is a potent, orally bioavailable small molecule tyrosine kinase inhibitor (TKI) against c-Met, VEGFR2 as well as Axl, Mer, and FLT3, all of which have been implicated in tumor pathogenesis. In cell-based assays, CT053PTSA showed IC50s of 6.7, 1.9 and < 1.0 nM against c-Met, VEGFR-2 and Axl. In cell-based functional assays, CT053PTSA inhibited HGF and VEGF-stimulated HUVEC proliferation and microvascular angiogenesis in rat aortic rings with IC50 values of 8.6 and 6.3 nM, respectively. When single dosed orally (3mg/kg) to U87MG tumor-bearing nude mice, CT053PTSA potently inhibited the phosphorylation of c-Met and its downstream signaling kinases AKT and ERK1/2 for up to 6 hours in tumor tissues.

The antitumor efficacy of CT053PTSA was evaluated in human tumor subcutaneous xenograft models in athymic mice, which included Caki-1 (kidney carcinoma), U-87MG (glioblastoma), NCI-H441 (lung adenocarcinoma), MDA-MB-231 (breast adenocarcinoma), SMMC-7721 (hepatoma), 5637 (bladder carcinoma) and SK-OV-3 (ovarian carcinoma). The ED50s of CT053PTSA to inhibit tumor growth in these studies ranged from < 1mg/kg (NCI-H441) to ∼ 6 mg/kg (SMMC-7721). In orthotopic U87MG human glioblastoma xenograft model, CT053PTSA prolonged the median survival time (MST) and yielded significant increase in life-span value (ILS = 32%, p = 0.003) at an oral dose of 20 mg/kg/day (dosed 21 days) versus the vehicle-treated group.

Conclusions: CT053PTSA displayed excellent oral bioavailability (> 60% across animal species) and desirable drug exposures in mice, rats, dogs and monkeys. Based on its potent antitumor activity, favorable PK profile and toxicological data, CT053PTSA was granted to enter clinical trials in China and is currently under phase I evaluation as a potential treatment for human malignancies.

Citation Format: Ning Xi, Yingjun Zhang, Zhaohe Wang, Yanjun Wu, Tingjin Wang. CT053PTSA, a novel c-MET and VEGFR2 inhibitor, potently suppresses angiogenesis and tumor growth. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1755. doi:10.1158/1538-7445.AM2014-1755
Abstract 1764: Inhibition of R-spondin (RSPO) signaling reduces the growth of multiple human tumors
Authors: Gurney Austin , Axelrod Fumiko , Bond Chris , Cain Jennifer , Chartier Cecile , Fischer Marcus , Ji May , Murriel Chris , Raval Janak , Shah Jalpa , Wang Min , Yen Wan-Ching , Kapoun Ann , Lewicki John , Hoey Timothy
Published: May 2015

Deregulation of the canonical Wnt/beta-catenin signaling pathway has long been associated with cancer. Intracellular components of this pathway, including Axin, APC, and beta-catenin are frequently mutated in a range of human tumors. The identity of specific extracellular ligands that contribute to human cancer development through this signaling axis has remained unclear. Molecular characterization of the secreted beta-catenin signaling activities produced by minimally passaged human tumor xenograft models identified RSPO family members produced by multiple tumor types including ovarian, pancreatic, colon, breast and non-small cell lung cancer. In human tumor xenograft models that had RSPO overexpression, in some instances due to genomic translocation, anti-RSPO treatment markedly inhibited tumor growth. In addition, striking combination activity with standard of care chemotherapy agents resulted in regression of established tumors. These results highlight the potential for therapeutic intervention with this newly appreciated signaling axis.

Citation Format: Austin Gurney, Fumiko Axelrod, Chris Bond, Jennifer Cain, Cecile Chartier, Marcus Fischer, May Ji, Chris Murriel, Janak Raval, Jalpa Shah, Min Wang, Wan-Ching Yen, Ann Kapoun, John Lewicki, Timothy Hoey. Inhibition of R-spondin (RSPO) signaling reduces the growth of multiple human tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1764. doi:10.1158/1538-7445.AM2014-1764
Abstract 1780: Identification and characterization of L524-0366 as a small molecule inhibitor that disrupt TWEAK-Fn14 signaling in glioblastoma
Authors: Dhruv Harshil D. , Loftus Joseph C. , Narang Pooja , Petit Joachim L. , Chow Donald , Yin Holly , Berens Michael , Meurice Nathalie , Tran Nhan L.
Published: May 2015

Deregulation of the TWEAK-Fn14 signaling pathway is observed in many diseases including inflammation, autoimmune diseases, and cancer, notably glioblastoma. Activation of Fn14 signaling by TWEAK binding triggers glioma cell invasion and survival and therefore represents an attractive pathway for therapeutic intervention. Based on structural studies of the TWEAK-binding cysteine rich domain of Fn14, several homology models of TWEAK were built to investigate plausible modes of TWEAK-Fn14 interaction. Two promising models, centered on different anchoring residues of TWEAK [Tyrosine 176 (Y176) and Tryptophan 231 (W231)], were prioritized using a data-driven strategy. Site-directed mutagenesis of TWEAK at Y176, but not W231, resulted in the loss of TWEAK binding to Fn14 substantiating Y176 as the anchoring residue. Importantly, mutation of TWEAK at Y176 did not disrupt TWEAK trimerization, but failed to induce Fn14 mediated nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling. The validated structural models were utilized in a virtual screen to design a targeted library of small molecules predicted to disrupt the TWEAK-Fn14 interaction. 129 small molecules were screened iteratively, with identification of molecules producing up to 37% inhibition of TWEAK-Fn14 binding. One specific compound, L524-0366 binds specifically to the Fn14 receptor and disrupt TWEAK-Fn14 interaction. Treatment of L524-0366 suppresses TWEAK-induced cell migration and survival in glioblastoma cells. In summary, we present a data-driven in silico study revealing key structural elements of the TWEAK-Fn14 interaction, followed by experimental validation, serving as a guide for the design of small molecule inhibitors of the TWEAK-Fn14 ligand-receptor interaction. Our results provide the foundation for further exploration utilizing L524-0366 as a small molecule inhibitor to the TWEAK-Fn14 signaling axis.

Citation Format: Harshil D. Dhruv, Joseph C. Loftus, Pooja Narang, Joachim L. Petit, Donald Chow, Holly Yin, Michael Berens, Nathalie Meurice, Nhan L. Tran. Identification and characterization of L524-0366 as a small molecule inhibitor that disrupt TWEAK-Fn14 signaling in glioblastoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1780. doi:10.1158/1538-7445.AM2014-1780
Abstract 1783: Puupehenol natural product inhibits STAT3 signaling and induces antitumor cell effects in vitro against human glioblastoma cells
Authors: Hilliard Tyvette S. , Chock Christopher , Williams Philip , Turkson James
Published: May 2015

Glioblastoma multiforme (GBM) is a malignant brain tumor that lacks effective therapeutics and has a high mortality rate. Constitutively-activated signal transducer and activator of transcription 3 (STAT3) protein is prevalent and implicated in GBM phenotype and represents a valid anti-GBM target. To discover novel STAT3 inhibitors suitable for GBM therapy, natural product extracts were investigated. Whole extracts from marine organisms were first screened for effects against cell viability using the human glioma model line, U251MG that harbors aberrantly-active STAT3 and normal fibroblasts that do not. The screening campaign identified the puupehenol, CC-1-2 from the whole extracts of leiodermatium sp as a bioactive compound that suppressed U251MG cell viability, with an IC50 4.1±0.1 µM, compared to an IC50 value of 7.3±0.3 µM against normal NIH3T3 mouse fibroblasts. Western blot analysis showed pSTAT3 levels in U251MG cells substantially decreased in a time-dependent manner in response to CC-1-2. Decreased pSTAT3 in treated cells occurred in parallel with partial suppression of both phospho- Janus kinase (Jak) 2 and pSrc levels and no induction of protein tyrosine phosphatase, PTP1B. Phosphorylated extracellular signal-regulated kinase (Erk) 1/2 levels also decreased in time-dependent manner, while pAkt remained unchanged in U251MG cells under the same treatment conditions. Furthermore, treatment of U251MG cells with CC-1-2 led to dramatic morphological changes that occurred in a dose- and time-dependent manner, compared to minimal changes observed for similarly-treated MCF-7 breast cancer cells that do not harbor aberrantly-active STAT3. Colony survival assay of U251MG cells revealed single treatment with 1-10 µM CC-1-2 decreased colony sizes and numbers in a dose-dependent manner, compared to moderate effects on MCF-7. In vitro wound-healing assay further showed reduced U251MG cell migration into the denuded area in response to CC-1-2 treatment. CC-1-2 further induced apoptosis in U251MG cells in a dose-dependent manner and suppressed the expression of c-Myc, Cyclin D1, and Bcl-2 levels, which are known STAT3-regulated genes. Studies together identify CC-1-2 as a natural product inhibitor of STAT3 that preferentially induces antitumor cell effects against human glioblastoma cells.

Citation Format: Tyvette S. Hilliard, Christopher Chock, Philip Williams, James Turkson. Puupehenol natural product inhibits STAT3 signaling and induces antitumor cell effects in vitro against human glioblastoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1783. doi:10.1158/1538-7445.AM2014-1783
Abstract 1786: Development and pharmacological properties of PEGylated glucuronide-auristatin linkers
Authors: Burke Patrick J. , Hamilton Joseph Z. , Hunter Joshua H. , Jeffrey Scott C. , Doronina Svetlana O. , Okeley Nicole M. , Meyer David W. , Senter Peter D. , Lyon Robert P.
Published: May 2015

Antibody-drug conjugates (ADCs) are an emerging therapeutic modality for the treatment of cancer, exemplified by the recent clinical success of brentuximab vedotin and ado-trastuzumab emtansine. The majority of ADCs currently in development include functionalities that are hydrophobic, resulting in increased plasma clearance. Consequently, there has been significant interest in generating new linkers that compensate for this potential liability. In an attempt to mitigate the pharmacokinetic impact of monomethylauristatin E (MMAE) when conjugated to an antibody, we prepared drug-linkers incorporating a polyethylene glycol (PEG) polymer. Initial work demonstrated that incorporation of a discrete PEG24 polymer into the cleavable β-glucuronide-MMAE linker system could either increase or decrease the plasma clearance of the resulting ADCs, depending upon the configuration of the PEG. When inserted into the drug-linker as a stretcher unit between the maleimide and the cleavage site, PEG24 elicited increased ADC plasma clearance. Conversely, incorporation of the PEG24 as a side chain from a modified lysine residue adjacent to the maleimide resulted in ADCs with slower clearance. These differences in ADC pharmacokinetics translated into corresponding differences in antitumor activity in two xenograft models. Thus, incorporation of PEG24 into ADC linkers impacts both pharmacokinetics and activity in a structure-dependent manner.

Citation Format: Patrick J. Burke, Joseph Z. Hamilton, Joshua H. Hunter, Scott C. Jeffrey, Svetlana O. Doronina, Nicole M. Okeley, David W. Meyer, Peter D. Senter, Robert P. Lyon. Development and pharmacological properties of PEGylated glucuronide-auristatin linkers. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1786. doi:10.1158/1538-7445.AM2014-1786
Abstract 179: Presence of primary cilia in cancer cells correlates with prognosis of pancreatic ductal adenocarcinoma
Authors: Emoto Katsura , Masugi Yohei , Yamazaki Ken , Effendi Kathryn , Tsujikawa Hanako , Kitago Minoru , Itano Osamu , Kitagawa Yuko , Sakamoto Michiie
Published: May 2015

Primary cilia are microtubule-based organelles that protrude from basal bodies and are involved in cell differentiation, sensory functions, and planar cell polarity. Especially, primary cilia have become popular as the place for signal transduction pathways including sonic hedgehog, Wnt, and platelet-derived growth factor pathways. Although there are many studies examining the roles of primary cilia in the fields of embryology and physiology, few such studies have been carried out in the field of oncology, and the role of primary cilia in cancer cells is poorly understood especially in human samples. In this study, we identified primary cilia by immunofluorescence analysis in which primary cilia were visualized as green rods labeled with anti-acetylated α-tubulin adjacent to basal bodies detected as red dots labeled with anti-γ-tubulin. Primary cilia were found in human pancreatic cancer cell lines (PANC-1 and CFPAC-1) and the number of them increased in the condition of serum starvation. As for clinical samples, primary cilia were found in cancer cells in 25 of 100 pancreatic ductal carcinoma patients [1-109 (median 8) primary cilia / 3 field pictures]. Most primary cilia in cancer tissue were observed in areas showing well differentiated glandular structures in clinical samples. Patients whose cancers were primary cilia positive had a higher frequency of lymph node metastasis than those whose cancers were primary cilia negative (p = 0.016). Univariate analysis demonstrated that tumor size (p = 0.009), tumor grade (p = 0.001), lymph node metastasis (p = 0.008), and the presence of primary cilia (p = 0.002) correlated with overall survival. Multivariate analysis found that tumor grade (p < 0.001) and the presence of primary cilia (p = 0.001) were independent prognostic indicators. In summary, we showed that pancreatic cancer cells can form primary cilia and that the presence of primary cilia is significantly associated with the prognosis of pancreatic ductal adenocarcinoma. The presence of primary cilia has the potential to become a new diagnostic tool and therapeutic target.

Citation Format: Katsura Emoto, Yohei Masugi, Ken Yamazaki, Kathryn Effendi, Hanako Tsujikawa, Minoru Kitago, Osamu Itano, Yuko Kitagawa, Michiie Sakamoto. Presence of primary cilia in cancer cells correlates with prognosis of pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 179. doi:10.1158/1538-7445.AM2014-179
Abstract 1798: Mechanistic study of NVP-CGM097: a potent, selective and species specific inhibitor of p53-Mdm2
Authors: Valat Thérèse , Masuya Keiichi , Baysang Frédéric , Albrecht Geneviève , Buschmann Nicole , Erdmann Dirk , Furet Pascal , Gabriel Tobias , Gessier François , Hofmann Francesco , Holzer Philipp , Kallen Joerg , Pissot-Solderman Carole , Stutz Stefan , Chène Patrick , Jeay Sébastien
Published: May 2015

An effective strategy to restore p53 activity in cancer cells containing wild type p53 is to inhibit the Mdm2-p53 protein-protein interaction (PPI). NVP-CGM097 is a novel PPI inhibitor under evaluation in a Phase I clinical trial. It binds to the p53 binding-site of the Mdm2 protein, disrupting the interaction between both proteins, leading to an activation of the p53 pathway.

The main biophysical and biochemical inhibitory characteristics of NVP-CGM097 are presented here. These include an affinity constant for Mdm2 in the nanomolar range and a selectivity of 3 orders of magnitude vs. Mdm4. The binding kinetics of NVP-CGM097 to Mdm2 are characterized by a high association rate constant (Kon =37 x 106 M-1.s-1) and a moderate dissociation rate constant (Koff =0.071 s-1). Additionally, NVP-CGM097 exhibits an 8-fold greater affinity for Mdm2 over Nutlin-3 due to a longer residence time of the Mdm2-inhibitor complex.

Moreover, biochemical studies have revealed the species specificity of NVP-CGM097 with human Mdm2 being inhibited more strongly than the dog, mouse or rat forms of the protein. This was confirmed in cellular assays where NVP-CGM097 treatment resulted in induction of p53 target gene expression (p21, PUMA and Mdm2) only in human, but not in dog, mouse or rat cell lines.

Citation Format: Thérèse Valat, Keiichi Masuya, Frédéric Baysang, Geneviève Albrecht, Nicole Buschmann, Dirk Erdmann, Pascal Furet, Tobias Gabriel, François Gessier, Francesco Hofmann, Philipp Holzer, Joerg Kallen, Carole Pissot-Solderman, Stefan Stutz, Patrick Chène, Sébastien Jeay. Mechanistic study of NVP-CGM097: a potent, selective and species specific inhibitor of p53-Mdm2. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1798. doi:10.1158/1538-7445.AM2014-1798