bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Sodium Alginate on Hypoglycemic Activity of Metformin Hydrochloride in the Microspheres Obtained by the Spray Drying
Authors: Szekalska Marta , Wróblewska Magdalena , Sosnowska Katarzyna , Winnicka Katarzyna
Published: May 2016

Alginate microspheres with metformin hydrochloride were prepared by the spray drying method in order to improve residence time of drug in the stomach. Nine formulations (F1–F9) with various drug : polymer ratio (1 : 2, 1 : 1, and 2 : 1) and different sodium alginate concentration (1%, 2%, and 3%) were evaluated for size, morphology, drug loading, Zeta potential, and swelling degree.In vitrodrug release, mathematical release profile, and physical state of microspheres were also evaluated. Optimal formulation characterized by the highest drug loading was formulation F6 (drug : polymer ratio 2 : 1 and 2% alginate solution). Based on glucose uptake inSaccharomyces cerevisiaecells andα-amylase inhibition tests, it could be concluded that alginate microspheres enhance hypoglycemic activity of metformin hydrochloride evaluatedin vitro. Designed microspheres are promising as alternative, multicompartment dosage form for metformin hydrochloride delivery.
Molecular Analysis of Vancomycin-Resistant Enterococci Isolated from Regional Hospitals in Trinidad and Tobago
Authors: Akpaka Patrick E. , Kissoon Shivnarine , Jayaratne Padman
Published: May 2016

Geographic spread of vancomycin-resistant enterococci (VRE) clones in cities, countries, or even continents has been identified by molecular techniques. This study aimed at characterizing virulent genes and determining genetic relatedness of 45 VRE isolates from Trinidad and Tobago using molecular tools, including polymerase chain reaction, pulsed-field gel electrophoresis (PFGE), and Random Amplification Polymorphic DNA (RAPD). The majority (84%) of the isolates wereEnterococcus faeciumpossessingvanA gene while the rest (16%) wereEnterococcus faecalispossessingvanB. Theespgene was found in all 45 VRE isolates whilehylgenes were found only inE. faeciumspecies. TheE. faeciumspecies expressed five distinct PFGE patterns. The predominant clones with similar or common patterns belonged to clones one and three, and each had 11 (29%) of the VRE isolates. Plasmid content was identified in representative isolates from each clonal group. By contrast, theE. faecalisspecies had one PFGE pattern suggesting the presence of an occult and limited clonal spread. The emergence of VRE in the country seems to be related to intra/interhospital dissemination of an epidemic clone carrying thevanA element. Therefore, infection control measures will be warranted to prevent any potential outbreak and spread of VRE in the country.
A Simulation and Validation of CLM during Freeze-Thaw on the Tibetan Plateau
Authors: Fang Xuewei , Luo Siqiong , Lyu Shihua , Chen Boli , Zhang Yu , Ma Di , Chang Yan
Published: May 2016

The applicability of a new soil hydraulic property of frozen soil scheme applied in Community Land Model 4.5 (CLM4.5), in conjunction with an impedance factor for the presence of soil ice, was validated through two offline numerical simulations conducted at Madoi (GS) and Zoige (ZS) on the Tibetan Plateau (TP). Sensitivity analysis was conducted via replacing the new soil hydraulic property scheme in CLM4.5 by the old one, using default CLM4.5 runs as reference. Results indicated that the new parameterization scheme ameliorated the surface dry biases at ZS but enlarged the wet biases which existed at GS site due to ignoring the gravel effect. The wetter surface condition in CLM4.5 also leads to a warmer surface soil temperature because of the greater heat capacity of liquid water. In addition, the combined impact of new soil hydraulic property schemes and the ice impedance function on the simulated soil moisture lead to the more reasonable simulation of the starting dates of freeze-thaw cycle, especially at the thawing stage. The improvements also lead to the more reasonable turbulent fluxes simulations. Meanwhile, the decreased snow cover fraction in CLM4.5 resulted in a lower albedo, which tended to increase net surface radiation compared to previous versions. Further optimizing is needed to take the gravel into account in the numerical description of thermal-hydrological interactions.
Lower Pectoralis Muscle Area Is Associated with a Worse Overall Survival in Non–Small Cell Lung Cancer
Authors: Kinsey C. Matthew , San José Estépar Raul , van der Velden Jos , Cole Bernard F. , Christiani David C. , Washko George R.
Published: May 2016

Background: Muscle wasting is a component of the diagnosis of cancer cachexia and has been associated with poor prognosis. However, recommended tools to measure sarcopenia are limited by poor sensitivity or the need to perform additional scans. We hypothesized that pectoralis muscle area (PMA) measured objectively on chest CT scan may be associated with overall survival (OS) in non–small cell lung cancer (NSCLC).

Methods: We evaluated 252 cases from a prospectively enrolling lung cancer cohort. Eligible cases had CT scans performed prior to the initiation of surgery, radiation, or chemotherapy. PMA was measured in a semi-automated fashion while blinded to characteristics of the tumor, lung, and patient outcomes.

Results: Men had a significantly greater PMA than women (37.59 vs. 26.19 cm2, P < 0.0001). In univariate analysis, PMA was associated with age and body mass index (BMI). A Cox proportional hazards model was constructed to account for confounders associated with survival. Lower pectoralis area (per cm2) at diagnosis was associated with an increased hazard of death of 2% (HRadj, 0.98; confidence interval, 0.96–0.99; P = 0.044) while adjusting for age, sex, smoking, chronic bronchitis, emphysema, histology, stage, chemotherapy, radiation, surgery, BMI, and ECOG performance status.

Conclusions: Lower PMA measured from chest CT scans obtained at the time of diagnosis of NSCLC is associated with a worse OS.

Impact: PMA may be a valuable CT biomarker for sarcopenia-associated lung cancer survival. Cancer Epidemiol Biomarkers Prev; 26(1); 38–43. ©2016 AACR.

See all the articles in this CEBP Focus section, “The Obesity Paradox in Cancer: Evidence and New Directions.”
Alcohol Use and Breast Cancer Survival among Participants in the Women's Health Initiative
Authors: Lowry Sarah J. , Kapphahn Kris , Chlebowski Rowan , Li Christopher I.
Published: May 2016

Background: Alcohol increases the risk of breast cancer even at moderate levels of intake. However, the relationship between alcohol consumption and mortality among breast cancer patients is less clear.

Methods: This study included women from the Women's Health Initiative observational study and randomized trial diagnosed with breast cancer (n = 7,835). Cox proportional hazards regression was used to estimate adjusted HRs and 95% confidence intervals (CI) for overall and breast cancer–specific (BCS) mortality associated with drinking alcohol before or after a breast cancer diagnosis. We also assessed whether changes in drinking habits after diagnosis are related to mortality.

Results: Women who were consuming alcohol prior to their breast cancer diagnosis had a nonstatistically significant 24% (95% CI, 0.56–1.04) reduced risk of BCS mortality and a 26% (95% CI, 0.61–0.89) reduced risk of all-cause mortality. Some variation was observed by estrogen receptor (ER) status as alcohol consumption was associated with a 49% (95% CI, 0.31–0.83) reduced risk of BCS mortality among ER− patients with no change in risk observed among ER+ patients (HR = 0.97; 95% CI, 0.31–1.54), though the difference between these risks was not statistically significant (P for interaction = 0.39). Postdiagnosis alcohol consumption, and change in consumption patterns after diagnosis, did not appear to be associated with all-cause or BCS mortality.

Conclusion: In this large study, consumption of alcohol before or after breast cancer diagnosis did not increase risks of overall or cause-specific mortality.

Impact: Coupled with existing evidence, alcohol consumption is unlikely to have a substantial impact on mortality among breast cancer patients. Cancer Epidemiol Biomarkers Prev; 25(8); 1268–73. ©2016 AACR.
Ultrastructural Insights into the Biological Significance of Persisting DNA Damage Foci after Low Doses of Ionizing Radiation
Authors: Lorat Yvonne , Schanz Stefanie , Rübe Claudia E.
Published: May 2016

Purpose: Intensity-modulated radiotherapy (IMRT) enables the delivery of high doses to target volume while sparing surrounding nontargeted tissues. IMRT treatment, however, substantially increases the normal tissue volume receiving low-dose irradiation, but the biologic consequences are unclear.

Experimental Design: Using mouse strains that varied in genetic DNA repair capacity, we investigated the DNA damage response of cortical neurons during daily low-dose irradiation (0.1 Gy). Using light and electron microscopic approaches, we enumerated and characterized DNA damage foci as marker for double-strand breaks (DSBs).

Results: During repeated low-dose irradiation, cortical neurons in brain tissues of all mouse strains had a significant increase of persisting foci with cumulative doses, with the most pronounced accumulation of large-sized foci in repair-deficient mice. Electron microscopic analysis revealed that persisting foci in repair-proficient neurons reflect chromatin alterations in heterochromatin, but not persistently unrepaired DSBs. Repair-deficient SCID neurons, by contrast, showed high numbers of unrepaired DSBs in eu- and heterochromatin, emphasizing the fundamental role of DNA-PKcs in DSB rejoining, independent of chromatin status. In repair-deficient ATM−/− neurons, large persisting damage foci reflect multiple unrepaired DSBs concentrated at the boundary of heterochromatin due to disturbed KAP1 phosphorylation.

Conclusion: Repeated low-dose irradiation leads to the accumulation of persisting DNA damage foci in cortical neurons and thus may adversely affect brain tissue and increase the risk of carcinogenesis. Multiple unrepaired DSBs account for large-sized foci in repair-deficient neurons, thus quantifying foci alone may underestimate extent and complexity of persistent DNA damage. Clin Cancer Res; 22(21); 5300–11. ©2016 AACR.
The Obama Administration's Cancer Moonshot: A Call for Proteomics
Authors: Conrads Thomas P. , Petricoin Emanuel F.
Published: May 2016

The Cancer Moonshot Program has been launched and represents a potentially paradigm-shifting initiative with the goal to implement a focused national effort to double the rate of progress against cancer. The placement of precision medicine, immunotherapy, genomics, and combination therapies was placed at the central nexus of this initiative. Although we are extremely enthusiastic about the goals of the program, it is time we meet this revolutionary project with equally bold and cutting-edge ideas: it is time we move firmly into the postgenome era and provide the necessary resources to propel and seize on innovative recent gains in the field of proteomics required for it to stand on equal footing in this narrative as a combined, synergistic engine for molecular profiling. After all, although the genome is the information archive, it is the proteins that actually do the work of the cell and represent the structural cellular machinery. It is the proteins that comprise most of the biomarkers that are measured to detect cancers, constitute the antigens that drive immune response and inter- and intracellular communications, and it is the proteins that are the drug targets for nearly every targeted therapy that is being evaluated in cancer trials today. We believe that a combined systems biology view of the tumor microenvironment that orients cancer studies back to the functional proteome, phosphoproteome, and biochemistry of the cell will be essential to deliver on the promise of the Cancer Moonshot Program. Clin Cancer Res; 22(18); 4556–8. ©2016 AACR.
Abstract A107: Cancer invadopodia controlled down-regulated miRNAs in esophageal squamous cell carcinoma
Authors: Kano Masayuki , Matsumoto Yasunori , Matsubara Hisahiro
Published: May 2016

Esophageal squamous cell carcinoma (ESCC) is one of the most malignant cancers of the gastrointestinal tract. However, molecular indicators of the origin of cellular deregulation in ESCC have not been identiﬁed. MicroRNAs (miRNAs), noncoding RNAs 21-25 nucleotides in length, regulate gene expression primarily at the posttranscriptional level. Growing evidence suggests that miRNAs are aberrantly expressed in many human cancers, and that they play significant roles in carcinogenesis and cancer progression. A search for miRNAs with a tumor-suppressive function in ESCC was performed using the miRNA expression signatures. miRNA signatures from ESCC identified miRNAs that are downregulated in common, and these miRNAs are candidate tumor suppressors. Gain-of-function analysis revealed that 3 transfectants (miR-145, miR-133a and miR-133b) inhibit cell proliferation and cell invasion. These miRNAs, which have conserved sequences in the 3'UTR of FSCN1, inhibited FSCN1 expression. The signal from a luciferase reporter assay was significantly decreased at 2 miR-145 target sites and 1 miR-133a/b site, suggesting both miRNAs directly regulate FSCN1. An FSCN1 loss-of-function assay found significant cell growth and invasion inhibition, implying an FSCN1 is associated with ESCC carcinogenesis. The identification of tumor-suppressive miRNAs, miR-145, miR-133a and miR-133b, directly control oncogenic FSCN1 gene. These signal pathways of ESCC could provide new insights into potential mechanisms of ESCC carcinogenesis.

FSCN1 is actin bundling protein, which is concerned with cancer invadopodia. Cancer invadopodia is one of the most important phenomenon regarding cancer cell invasion into surrounding tissue. Many kinds of molecules participate invadopodia in the cancer cell. We focused on MMP14 and examined the gene expression of MMP14 in ESCC tissues and patient survival. FSCN1 and MMP14 were evaluated by immunohistochemistry and quantitative PCR, and microRNA (miR)-133a was also evaluated by PCR in surgical ESCC specimens. The roles of FSCN1, MMP14 and miR-133a were established in ESCC cells. The relationship between the expressions of miR-133 and MMP14 is strongly negative correlation. miR-145 and miR-133 play a chief role in cancer invasion and metastasis, which control invadopodia associated proteins.

The combined expression of FSCN1 and MMP14 is associated with a poor prognosis, and miR-133a, which regulates their mRNAs, can serve as a strong tumour suppressor of ESCC.

Citation Format: Masayuki Kano, Yasunori Matsumoto, Hisahiro Matsubara. Cancer invadopodia controlled down-regulated miRNAs in esophageal squamous cell 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 A107.
Abstract A130: Targeting BMI-1 by the novel BMI-1 inhibitor PTC596 in acute leukemia
Authors: Nishida Yuki , Maeda Aya , Cao Liangxian , Dumble Melissa , Kimura Shinya , Davis Thomas W. , Kojima Kensuke
Published: May 2016

Leukemia stem cells are resistant to chemotherapeutic agents and contribute to disease relapse. BMI-1, a part of polycomb repressive complex 1 (PRC1) is essential for the self-renewal of normal hematopoietic and leukemia stem cells. We have reported that acute myeloid leukemia (AML) patients with higher levels of BMI-1 have worse overall survival and that the translational BMI-1 inhibitor PTC-209 induces apoptosis in patient-derived CD34+CD38low/- AML cells. PTC596 is a second-generation BMI-1 inhibitor that accelerates BMI-1 degradation. Based on preclinical data showing excellent anti-tumor activities against solid cancers, PTC596 is currently in PH1 clinical development. We here investigated preclinical activities of PTC596 against acute leukemias in vitro and in vivo. A total of 6 AML (MOLM-13, OCI-AML3, U-937, MV4-11, MOLM-14 and HL60) and 3 acute lymphoblastic leukemia (ALL) (Reh, NALM6 and MOLT-4) cell lines were exposed to PTC596 for 48 hours. PTC596 exhibited dose- and time-dependent anti-proliferative and cytotoxic (apoptotic) activities. The IC50 values (concentration at which cell growth is inhibited by 50% at 48 hours of exposure) were 30.7 ± 4.1 nM (mean ± SEM) for AML and 37.7 ± 8.0 nM for ALL. The ED50 values (effective concentration inducing 50% killing as measured by Annexin V/PI induction) were 60.3 ± 6.7 nM for AML and 77.8 ± 11.5 nM for ALL. PTC596 induced apoptosis in AML cells irrespective of their p53 status. 72-hour treatment of MOLM-13 and OCI-AML3 cells with 100nM PTC596 reduced total BMI-1 protein levels by 87% and 61%, respectively. Forced overexpression of BMI-1 in K562 cells abrogated PTC596-induced apoptosis (∼ 80% at 50 nM PTC596; P < 0.05). 16-hour treatment of MOLM-13 cells with 200nM PTC596 resulted in reduced levels of non-phosphorylated BMI-1 by 85%, along with increased levels of phosphorylated BMI-1. PTC596 reduced the levels of ubiquitylated histone H2A, which indicates a deactivation of the PRC1 complex function. The anti-leukemia activity of PTC596 was tested in xenograft (K562) and syngeneic (L210) mouse models. PTC596 at a daily dose of 20 mg/kg significantly delayed an enlargement of the implanted tumor of K562 by ∼55%. PTC596 significantly prolonged survival of mice injected with syngeneic L210 leukemia cells (27 versus 37 days; 76% extension of lifespan with a median survival time; P < 0.05). Collectively, BMI-1 inhibition by PTC596 offers a novel therapeutic strategy for acute leukemias.

Citation Format: Yuki Nishida, Aya Maeda, Liangxian Cao, Melissa Dumble, Shinya Kimura, Thomas W. Davis, Kensuke Kojima. Targeting BMI-1 by the novel BMI-1 inhibitor PTC596 in acute leukemia. [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 A130.
Abstract A144: Restoration of tumor suppressor function by mitochondrially targeting p14 peptide in tumor cells
Authors: Saito Ken , Kondo Eisaku
Published: May 2016

Introduction

Loss of CDKN2A (encoding p16INK4a and p14ARF) gene mainly by epigenetic alternations is associated with an increased susceptibility for development of malignant tumors. We previously reported the loss of the p14ARF function especially on the gefitinib-resistant lung adenocarcinomas (NSCLCs). In the present study, we identified the p14MIS (p14 minimal inhibitory sequence) peptide covering the functional core sequence of p14ARF, which is targeted to the mitochondria in the tumor cells and report its utility in growth suppression of biologically aggressive cancer cells of diverse origins.

Experimental procedures

Human tumor lines of various origins were treated with 10 μM of the p14MIS for 24 h or 48 h. The intracellular localization of p14MIS and growth suppression which is correlated to mitochondrial membrane potential by the p14MIS were examined by MTT assay and inverted fluorescence microscopy which is available to the peptide-treated living tumor cells. For in vivo study, the tumor xenograft mouse models were made by intraperitoneal injection of the human pancreatic adenocarcinoma cell, GFP-expressing BxPC3, and examined the antitumor effect of the peptide. The mice were daily treated with the 300ug/mouse of the p14MIS or with the polyarginine peptide (R9) as a control for a week, respectively. After the sequential treatment of the peptide, the mice of each group were autopsied and evaluated intraperitoneal dissemination (metastasis) of the GFP-expressing tumors.

Results

The p14MIS successfully targeted to the mitochondria and triggered an apoptosis to the various lineage of tumor cells by reduction of their mitochondrial membrane potential. The sensitivity and efficacy of the p14MIS was dependent on the expression level of endogenous mitochondrial ATPAF1 (F1-ATPase assembly protein) and also the magnitude of downregulation of the mitochondrial membrane potential in each tumor line. Moreover, in vivo delivery of the p14MIS to the pancreatic tumor model mice resulted in a 75% reduction in tumor volume compared with those in the control mice.

Conclusions

These results support the biological significance of the tumor suppressor molecule, p14ARF, especially localized at mitochondrias on tumor cells, and the utility of p14MIS peptide as an antitumor biotool against highly aggressive cancers. Although molecular details in tumor cells still remain to be clarified with respect to mitochondrial action of the p14MIS, it a potential of the peptide-based molecular therapy in intractable human malignancies of diverse origins.

Citation Format: Ken Saito, Eisaku Kondo. Restoration of tumor suppressor function by mitochondrially targeting p14 peptide in tumor cells. [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 A144.
Abstract A151: Quantitative systems pharmacology approaches accelerate lead generation and optimization of a PD-1 x TIM-3 therapeutic in immuno-oncology
Authors: Apgar Joshua F. , Wong Jamie , Ryan Phennicie Ryan , Briskin Mike , Burke John M.
Published: May 2016

The goal of this collaboration was to provide early quantitative decision making guidance for the project team by developing and interrogating a quantitative systems pharmacology (QSP) model of the co-modulation inhibitory receptors PD-1 and TIM-3 in immuno-oncology. The QSP model was to: (1) provide predictions of the best-in-class profile for a PD-1 and TIM-3 dual antagonist, (2) accelerate project timelines, (3) provide biological insights, and (4) reduce experimental costs.

The QSP model was based on first principles as a system of elementary mass-action, mechanistic PKPD, ordinary differential equations. The model parameters and reactions were based on biophysics, and are interpretable. The model reactions include protein synthesis and elimination, ligand-receptor and drug-target formation and turnover, and drug administration and first order clearance. There were four versions of the model: PD-1 monospecific, TIM-3 monospecific, PD-1 x TIM-3 bispecific and fixed dose combination (FDC) targeting PD-1 and TIM-3. The monospecific models were then benchmarked against published data such that model parameter values were set to known values and unknown parameters were estimated. Once benchmarked, the FDC and bispecific models were analyzed by systematically investigating how tuning the model parameters (e.g., affinity, avidity, dose, half-life, target expression, etc.) impacted target inhibition, and to simulate patient variability.

The model was in good agreement with published clinical data from nivolumab and pembrolizumab, and data from RMT3-23 in the TIM-3 driven mouse model. QSP model analysis predicted: (1) there would be diminishing returns on very tight binding biologics due to Target Mediated Drug Disposition (TMDD) that offsets potency, and (2) there is no advantage between FDC, 2-2 bispecific, and 2-1 bispecific formats, which are predicted to be roughly equivalent. As a result of these analyses, there was a significant reduction in the number of experiments, and acceleration of project timelines by (1) eliminating rounds of affinity maturation, as drug leads were in predicted optimal drug parameter ranges, and (2) eliminating the need to construct and evaluate bi-specific constructs and proceed with FDCs.

Citation Format: Joshua F. Apgar, Jamie Wong, Ryan Ryan Phennicie, Mike Briskin, John M. Burke. Quantitative systems pharmacology approaches accelerate lead generation and optimization of a PD-1 x TIM-3 therapeutic in immuno-oncology. [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 A151.
Abstract A167: A phase I study of MET TKI SAR125844 in Asian patients (pts) with advanced solid tumors
Authors: Shitara Kohei , Oh Do-Youn , Yokota Tomoya , Kim Tae Won , Doi Toshihiko , Hamauchi Satoshi , Ahn Jin-Hee , Gomez Corinne , Harnois Marzia , Jue Bang Yung
Published: May 2016

Background: SAR125844 (SAR) is a potent and small molecule inhibitor of the MET kinase (IC50 = 4.2nM; Ki = 2.8nM). This phase I study (NCT01657214) was initiated to address the maximum tolerated dose (MTD), safety, pharmacokinetics (PK), pharmacodynamics (PD) and preliminary antitumor activity in adult Asian pts with advanced solid tumors.

Methods: Eligible pts received SAR by IV infusion every week. Dose escalation proceeded with an adaptive Bayesian method. At the MTD, pts with advanced c-Met amplified solid tumors including gastric cancer (GC) pts were enrolled in an expansion cohort.

Results: As of 30 June 2015, 32 pts have received SAR, 19 pts in the dose escalation phase over 4 dose levels (DL) ranging from 260 to 570 mg/m2 and 13 pts in the MTD expansion cohort including 8 pts with advanced GC. Of the 32 treated pts, median age was 64 years [28-78], 18M/14F, ECOG-PS 0/1/2: 14/16/2 with various solid tumors including 16 GC, 6 colorectal and 4 lung adenocarcinomas. Pts received a median of 4 infusions [2-20]. No dose-limiting toxicity (DLT) was reported nor MTD was reached. Of the 19 pts treated in dose escalation, blood exposure PK parameters (AUC and Cmax) increased in proportion with the dose with a mean clearance of 34 L/h associated with a large volume of distribution (239 L). Dose expansion was opened at 570 mg/m2 for pts with solid tumor and c-Met-gene amplification determined on local FISH assay. The most commonly reported all-causality AE (≥15% of pts) were: nausea, vomiting, decreased appetite, asthenia/fatigue, constipation, pyrexia, abdominal pains, injection site reaction and back pain. 31.3% of pts had grade ≥3 AEs, none were related to study drug. No relevant severe laboratory abnormality was observed. In the dose escalation part in unselected pts, stable disease (SD) was reported in 8 pts at the 4 week assessment. Of the 13 pts treated so far in dose expansion, 2 pts with GC achieved confirmed partial response and 5 pts had SD (4GC and 1sarcoma).

Conclusions: SAR administered as single agent at the dose of 570mg/m2 has a tolerable and manageable safety profile and encouraging anti-tumor activity is seen in pts with c-Met -amplified GC. Accrual of pts with c-Met-gene amplified GC is still ongoing.

Sponsored by Sanofi; NCT01657214

Citation Format: Kohei Shitara, Do-Youn Oh, Tomoya Yokota, Tae Won Kim, Toshihiko Doi, Satoshi Hamauchi, Jin-Hee Ahn, Corinne Gomez, Marzia Harnois, Yung Jue Bang. A phase I study of MET TKI SAR125844 in Asian patients (pts) with advanced solid tumors. [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 A167.
Abstract A168: Discovery of exquisitely selective MerTK inhibitors
Authors: Yoon Taeyoung , Sheen Joon-Ho , Park Seong Jin , Kim Jin Kwan , Kim Hadong , Kim Eun-Jung , Kwon Munjin , Lee Hyung Ki , Lee Okyung , Jeong Youngjee , Ryu Ki Moon , Jang Miyeon , Hwang Jae-Sung
Published: May 2016

MerTK is a receptor tyrosine kinase most prominently expressed in myeloid cells and known for its role in phagocytosis of early apoptotic cells and concomitant establishment of tolerogenic microenvironment. Hence, MerTK is thought to play a significant role in cancer immune evasion as, for example, a defining component of M2c-type tumor-associated macrophages (TAMs). Moreover, it has also been increasingly recognized that MerTK is overexpressed in a variety of malignant tumor cells, promoting the cancer cell survival, invasion, and treatment resistance. As such, it is anticipated that the cancer cell intrinsic and extrinsic effects might conspire to make MerTK a highly promising target for anticancer therapy. We describe herein an exquisitely selective series of MerTK inhibitors. A lead compound, XL1547, exhibited an IC50 of 1 nM against MerTK in an enzyme assay, with a greater than 300-fold selectivity over closely related AXL kinase (IC50 = 350 nM). In fact, MerTK was the only enzyme that was inhibited by XL1547 (> 99% at 100 nM) in the panel of 386 kinases (Reaction Biology Corp.; all others < 35%). X-ray crystal structure revealed a unique binding mode where the compound exploits a large back pocket next to the gatekeeper residue (known as a Type 1.5 binder). XL1547 potently inhibited autophosphorylation as well as the viability of engineered BaF3(CD8:MerTK) cells (GI50 = 230 nM) with little toxicity. Extensive medicinal chemistry efforts yielded a more advanced compound SA3686 with at least 10-fold improvement in potency. More importantly, it displayed a pharmacokinetic profile suitable for oral dosing. Indeed, SA3686 was highly efficacious in vivo in an allograft tumor model of subcutaneously implanted Ba/F3(CD8:MerTK) cells. [SA3686 is being tested in multiple other in vivo models at the time of abstract submission]. In summary, we have identified an essentially mono-selective MerTK inhibitor series that will help delineate the biology of MerTK-specific inhibition and hopefully validate the translational potential of MerTK as a therapeutic target for human diseases including cancer.

Citation Format: Taeyoung Yoon, Joon-Ho Sheen, Seong Jin Park, Jin Kwan Kim, Hadong Kim, Eun-Jung Kim, Munjin Kwon, Hyung Ki Lee, Okyung Lee, Youngjee Jeong, Ki Moon Ryu, Miyeon Jang, Jae-Sung Hwang. Discovery of exquisitely selective MerTK inhibitors. [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 A168.
Abstract A172: Preclinical antitumor activity of ASP5878, a novel inhibitor of FGFR1, 2, 3 and 4, in FGF19-expressing hepatocellular carcinoma
Authors: Futami Takashi , Okada Hidetsugu , Kihara Rumi , Kawase Tatsuya , Nakayama Ayako , Suzuki Tomoyuki , Kameda Minoru , Shindoh Nobuaki , Terasaka Tadashi , Hirano Masaaki , Kuromitsu Sadao
Published: May 2016

Background: Fibroblast growth factor (FGF) / FGF receptor (FGFR) gene aberrations such as amplification, mutation and fusion are associated with many types of human cancers. Recently, FGF19 overexpression was observed in approximately 50% of hepatocellular carcinoma (HCC) patients. The FGF19-FGFR4 signaling has been implicated in the development of HCCs in mice. FGFR4 kinase inhibitors are expected to be a targeted therapy for FGF19-expressing HCC. A phase I clinical trial of ASP5878, a novel inhibitor of FGFR1, 2, 3 and 4, is ongoing (NCT02038673). Method: We tested selectivity of ASP5878 among 128 kinases and sensitivity of ASP5878 on cell proliferation of HCC cell lines. Activation of FRS2 and ERK, downstream molecules of FGFR signaling, and PARP cleavage in FGF19 expressing HCC cell lines were evaluated with Western blotting. In vivo antitumor effects of ASP5878 were examined in HCC subcutaneous xenograft and orthotopic inoculation mouse models. Finally, plasma levels of FGF19 were measured after dosing ASP5878. Results: Among 128 kinases, only 9 kinases including FGFR1-4 and FGFR3/4 mutations were inhibited more than 50% by ASP5878 (200 nmol/L). The IC50 values of ASP5878 against FGFR1, 2, 3 and 4 kinases were 0.47, 0.60, 0.74 and 3.5 nmol/L, respectively. ASP5878 inhibited cell proliferation of HCC cell lines with FGF19 overexpression. IC50 values were 8.5, 27, and 21 nmol/L in Hep3B2.1-7, HuH-7 and JHH-7, respectively. ASP5878 inhibited activation of downstream signaling molecules, FRS2 and ERK, and induced apoptosis in Hep3B2.1-7 cells. Oral dosing of ASP5878 at 3 mg/kg induced sustained tumor regression in the Hep3B2.1-7 subcutaneous xenograft model, which was poorly responsive to sorafenib. In an HuH-7 orthotopic inoculation mouse model, ASP5878 induced complete tumor regression and dramatically extended the survival. In addition, oral dosing of ASP5878 reduced plasma levels of FGF19 in the HuH-7 subcutaneous xenograft model.Conclusion: These results suggest that ASP5878 is a potentially effective therapeutic agent for FGF19-expressing HCC.

Citation Format: Takashi Futami, Hidetsugu Okada, Rumi Kihara, Tatsuya Kawase, Ayako Nakayama, Tomoyuki Suzuki, Minoru Kameda, Nobuaki Shindoh, Tadashi Terasaka, Masaaki Hirano, Sadao Kuromitsu. Preclinical antitumor activity of ASP5878, a novel inhibitor of FGFR1, 2, 3 and 4, in FGF19-expressing 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 A172.
Abstract A25: Predictive biomarkers for bevacizumab response in recurrent glioblastoma patients
Authors: Urup Thomas , Michaelsen Signe Regner , Olsen Lars Rønn , Toft Anders , Christensen Ib Jarle , Grunnet Kirsten , Broholm Helle , Kosteljanetz Michael , Issazadeh-Navikas Shohreh , Poulsen Hans Skovgaard , Lassen Ulriik
Published: May 2016

BACKGROUND: Bevacizumab (BEV) plus chemotherapy has shown high response rates in recurrent glioblastoma (GBM) and patients who achieve response have an improved overall survival as well as quality of life. Recent retrospective analysis of the randomized phase III trial, AVAglio, indicate that patients with the proneural GBM subtype have a survival benefit when treated with BEV in combination with standard treatment. However, no validated biomarkers able to predict BEV response have been identified and the biology reflecting a clinical BEV response is poorly understood. The primary objective of this study was to evaluate the predictive and prognostic value of GBM subtypes in recurrent GBM patients treated with BEV therapy. The secondary objective was to identify biomarkers able to predict response to BEV therapy in recurrent GBM patients. METHODS: A total of 90 recurrent GBM patients treated with BEV combination treatment according to a previously published treatment protocol were included. Inclusion criteria: BEV plus irinotecan treatment in the period between May 2005-2011; available GBM tissue (according to WHO); response evaluable (RANO). RNA from tumor tissue was analyzed by the NanoString platform covering 800 genes. Raw data was assigned to molecular subtypes for each of the samples using the PAMR classifier model, previously trained on the AVAglio dataset. In order to identify novel candidate biomarkers able to predict response, differentially expressed genes (fold-change difference > 1.5) between patients responding versus progressing on BEV were identified using a t-test. Biomarkers significantly (P<0.05) associated with response in multivariate logistic regression analysis adjusted for recently validated clinical prognostic factors were selected for the final predictive model. RESULTS: Molecular subtypes were not associated with response or overall survival. However, two novel independent predictive biomarkers (gene1 down-regulated and gene2 up-regulated in responders) of BEV response and overall survival were identified. Results will be presented.

Citation Format: Thomas Urup, Signe Regner Michaelsen, Lars Rønn Olsen, Anders Toft, Ib Jarle Christensen, Kirsten Grunnet, Helle Broholm, Michael Kosteljanetz, Shohreh Issazadeh-Navikas, Hans Skovgaard Poulsen, Ulriik Lassen. Predictive biomarkers for bevacizumab response in recurrent glioblastoma patients. [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 A25.
Abstract B109: AZD5363, a catalytic pan-Akt inhibitor, in Akt1 E17K mutation positive advanced solid tumors
Authors: Hyman David M. , Smyth Lillian , Bedard Philippe L. , Oza Amit , Dean Emma , Armstrong Anne , Lima Joao , Bando Hideaki , Kabos Peter , Perez-Fidalgo J. Alejandro , Moore Kathleen , Westin Shannon N. , You Benoit , Chandarlapaty Sarat , Alland Leila , Ambrose Helen , Foxley Andrew , Lindemann Justin , Pass Martin , Rugman Paul , Salim Shaista , Schiavon Gaia , Tamura Kenji , Baselga Jose , Banerji Udai
Published: May 2016

This abstract has been withheld from publication due to its inclusion in the AACR-NCI-EORTC Molecular Targets Conference 2015 Official Press Program. It will be posted online at the time of its presentation in a press conference or in a session: 10:00 AM ET Saturday, November 7.

Citation Format: David M. Hyman, Lillian Smyth, Philippe L. Bedard, Amit Oza, Emma Dean, Anne Armstrong, Joao Lima, Hideaki Bando, Peter Kabos, J. Alejandro Perez-Fidalgo, Kathleen Moore, Shannon N. Westin, Benoit You, Sarat Chandarlapaty, Leila Alland, Helen Ambrose, Andrew Foxley, Justin Lindemann, Martin Pass, Paul Rugman, Shaista Salim, Gaia Schiavon, Kenji Tamura, Jose Baselga, Udai Banerji. AZD5363, a catalytic pan-Akt inhibitor, in Akt1 E17K mutation positive advanced solid tumors. [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 B109.
Abstract B139: Determination of the plasma inhibitory activity of drugs using cellular kinase assays
Authors: Boehmer Kira E. , Weber Holger , Feger Daniel , Birkle Marianne , Siedentopf Oliver , Mueller Melanie , Umber Sarah , Ehlert Jan E.
Published: May 2016

Anticipation of the in vivo activity of new investigational drugs is challenging. In the context of the living body, compounds are subjected to degradation, modification or binding to plasma proteins - mechanisms that may compromise the activity predicted from target-specific in vitro assays. As such assays are performed in artificial cell-culture media it was our aim to measure the activity directly in human plasma, as initially described by Levis et al.1. We hence established a method to analyze the activity of kinase inhibitors in human or rodent plasma (plasma inhibitory activity, PIA) in high throughput compatible ELISA-based cellular kinase assays for e.g. FLT-3 or BCR-Abl. Testing reference compounds such as Sunitinib or Sorafenib, we could confirm the expected correlation between the plasma binding of a compound and the loss of its activity in this assay setup compared to the activity in serum free medium. Furthermore we observed a donor dependent interference of the plasma with assay performance. This issue was solved by appropriate plasma heat-inactivation which made confirmation of compound stability to heat treatment mandatory before generating pharmacokinetic data using this methodology. Our aim is to establish the PIA assay for our panel of more than 30 kinases and analyze the activity of important reference kinase inhibitors spiked in human plasma. In the next step samples from mice treated with reference compounds shall be tested. This way, we want to compile reliable in vitro PIA assays to better predict the in vivo effect of kinase-directed drugs and to determine the amount of freely available active test substance in the blood of treated patients.

1 = Levis et al.; Blood, 2006; 108:3477

Citation Format: Kira E. Boehmer, Holger Weber, Daniel Feger, Marianne Birkle, Oliver Siedentopf, Melanie Mueller, Sarah Umber, Jan E. Ehlert. Determination of the plasma inhibitory activity of drugs using cellular kinase assays. [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 B139.
Abstract B14: ALT specific C-circle detection in a whole blood of cancer patients using qPCR assay
Authors: Malykh Andrei , Doyle Jonathan
Published: May 2016

The goal of this study was to evaluate a real-time qPCR assay, which can be used in clinical research environments for detection of telomeric C-circle DNA in tumors and peripheral blood of patients with cancers that utilize Alternative Lengthening of Telomeres (ALT) as their Telomere length Maintenance Mechanism (TMM). Telomeric C-circle DNA has been demonstrated to be specific for ALT, thus enabling the first quantitative test for ALT activity. In 85% of cancers, cells maintain telomere length due to increased activity of telomerase, while 5 - 15% of cancers overall rely on the ALT mechanism for continued growth and survival. Initial retrospective analyses showed that ALT ranges from 25% to 60% in sarcomas and 5% to 15% in carcinomas. ALT is also a potential ‘escape route’ for tumor cells to survive under pressure from treatment with anti-telomerase drugs. Results of in vitro and animal model experiments confirm the possibility of such ALT induction.

The existing detection method using a radioactive probe was shown to detect C-circle DNA in osteosarcoma patients' blood and plasma. However, this method requires highly experienced technicians and dedicated laboratory facilities for handling radioisotopes. Here we present data that the qPCR assay is at least 20 times more sensitive than its radioisotope predecessor and detects C-circle DNA in blood of sarcoma patients.

DNA from 20 frozen whole blood and serum samples from patients with adenocarcinoma (10), sarcoma (4), desmoid fibrosarcoma (1) and melanoma (5) was isolated using QIAmp Kit. Rolling circle amplification of telomere C-circle DNA, which is the core of the assay, was done with and without Phi29 polymerase. Half of the reaction was subjected to radioisotope detection and half for qPCR. Positive results in both isotope and qPCR assay have been confirmed for 3 sarcoma samples. Two serum samples (one from adenocarcinoma and one from melanoma patient) were positive in qPCR, while negative in isotope assay.

A clinically applicable qPCR C-circle assay, which can also be used as a noninvasive blood-based test, will be suitable for ALT activity detection in cancer, as a companion diagnostic tool for anti-telomerase therapy and prognosis, and for anti-ALT and anti-telomerase drug development.

This study was partially supported by the NIH SBIR Grant 1R43CA168124-01A1.

Citation Format: Andrei Malykh, Jonathan Doyle. ALT specific C-circle detection in a whole blood of cancer patients using qPCR assay. [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 B14.
Abstract B191: Preclinical assessment of targeting the PI3K/AKT pathway in combination with other signal transduction pathway inhibitors
Authors: Lee Sang Hyun , Koblish Holly , Liu Xiangdong , Ye Haifen , Li Jun , Liu Xuesong , Diamond Melody , Zhang Yue , Hall Leslie , Dostalik Valerie , Favata Margaret , Behshad Elham , Wynn Richard , Li Yun-long , Combs Andrew , Xue Chu-Bio , Yao Wenqing , Hollis Gregory , Huber Reid , Scherle Peggy
Published: May 2016

Inhibitors targeting the phosphoinositide 3-kinase (PI3K)/serine-threonine protein kinase B (AKT) pathway have been developed and have shown efficacy in inhibiting tumor growth both in preclinical models and in the clinic. The effectiveness of PI3K/AKT inhibitors, however, is attenuated by the activation of alternative signaling pathways. Conversely, the PI3K/AKT pathway is considered to be a resistance mechanism for other therapeutic or cytotoxic agents. Therefore, understanding these mechanisms may help to inform the most optimal combinations for advancement into clinical studies. Here, we explored the potential of targeting the JAK/STAT or PIM signaling pathways in combination with a novel, potent and selective AKT inhibitor, INCB047775. Consistent with published data, pharmacological inhibition of AKT by INCB047775 caused the upregulation of PIM kinases. The combined therapies of PIM inhibition and AKT inhibition exhibited significant synergistic anti-tumor effects. Combination of PIM inhibition with AKT inhibition resulted in synergistic induction of apoptosis in cancer cells. Similarly, the combination of JAK tyrosine kinase inhibition with INCB047775 exerted synergistic inhibition of the proliferation of multiple myeloma cell lines. Similar data were observed with an inhibitor of PI3Kδ in combination with both PIM and JAK inhibition, demonstrating significant crosstalk between the PI3K/AKT pathway and other critical signaling pathways. These data provide the rationale for combinatorial therapies with inhibitors targeting JAK or PIM kinases with PI3K/AKT pathway inhibitors in the clinic.

Citation Format: Sang Hyun Lee, Holly Koblish, Xiangdong Liu, Haifen Ye, Jun Li, Xuesong Liu, Melody Diamond, Yue Zhang, Leslie Hall, Valerie Dostalik, Margaret Favata, Elham Behshad, Richard Wynn, Yun-long Li, Andrew Combs, Chu-Bio Xue, Wenqing Yao, Gregory Hollis, Reid Huber, Peggy Scherle. Preclinical assessment of targeting the PI3K/AKT pathway in combination with other signal transduction pathway inhibitors. [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 B191.
Abstract B44: Druggability assessment of protein tyrosine phosphatase binding site
Authors: Ghattas Mohammad A. , Raslan Noor , Sadeq Asil , Atatreh Noor
Published: May 2016

Protein tyrosine phosphatases play important roles in the signalling system in our bodies and were observed to be overexpressed in many diseases. For instance, PTP1B, the most famous member of this family, has a well-established role in diabetes mellitus, obesity and cancer. However, no PTP inhibitors have reached the market so far. This raised a question about the druggability of their catalytic site; which appears to be hydrophilic in nature and is believed to favour mainly polar ligands to bind with. In this study, we did an extensive data minding in the protein data bank to obtain all available crystal structures of all members in the PTP family. We analysed these crystal structures using SiteMap/Schrodinger. This software finds cavities in the examined proteins structure and evaluates these pockets according to their suitability for binding with drug-like ligands. Consequently, we classified PTPs according to their druggability scores; a representative of each category is shown and discussed accordingly. The influence of active site flexibility (open or closed) on PTPs druggability was studied. The effect of tight-bound water molecules on active site druggability was also assessed. This analysis can prove useful in the discovery of new PTP inhibitors and can help researchers to save unnecessary cost, money and efforts by prioritizing PTP targets according to which members have the most druggable catalytic pocket.

Citation Format: Mohammad A. Ghattas, Noor Raslan, Asil Sadeq, Noor Atatreh. Druggability assessment of protein tyrosine phosphatase binding site. [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 B44.