bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase I experience with an anti-glycotope monoclonal antibody, RAV12, in recurrent adenocarcinoma
Authors: Burris H. A. , Lewis N. , Rosen L. S. , Jones S. F. , Cohen R. B. , Kunkel L. A. , Stewart S. J.
Published: March 2020

14017

Background: RAV12 is a high affinity, internalizing, chimeric IgG1 monoclonal antibody (MAb) that binds RAAG12, a novel primate-restricted N-linked carbohydrate epitope present on multiple cell-surface proteins. RAAG12 is variably expressed on normal non-keratinizing epithelia, particularly those of GI origin (cytoplasm and apical membrane), and is not expressed on human tissues from the cardiovascular, endocrine, hematolymphatic, neuromuscular, and central nervous systems. IHC studies demonstrated diffuse membrane staining on > 90% of human colorectal, gastric, and pancreatic adenocarcinoma samples. RAAG12 is distinct from previously reported mucin or Lewis blood group carcinoma-associated carbohydrates. In vitro, RAV12 kills tumor cells by oncosis and facilitates ADCC. Methods: In December 2004, a phase I dose-escalation safety and pharmacokinetics (PK) trial was begun in patients with recurrent (1 to 3 prior treatments) adenocarcinoma. RAV12 was administered weekly x 4, twenty-eight patients have received treatment in the following cohorts: 0.3 mg/kg qw (6), 1.0 mg/kg qw (8), 1.5 mg/kg qw (7), 0.5 mg/kg biw (3), 0.75 mg/kg biw (2), and 0.5 mg/kg tiw (2). Sixteen patients had colorectal, 5 gastroesophageal, 4 pancreatic, 2 lung cancer, and 1 breast cancer. Responses were evaluated on day 42. Results: Preliminary analysis demonstrated dose-dependent PK. Three clinical syndromes associated with drug administration have been observed to date: 1) abdominal cramping pain (21 patients) and diarrhea (18 patients) particularly at the higher doses, 2) asymptomatic, self-limited, generally rapidly reversible excursions of liver function tests (16 patients), and 3) asymptomatic, self-limited, rapidly reversible excursions of pancreatic enzymes (6 patients). One patient (0.3 mg/kg qw), with advanced pancreatic cancer, had a > 50% reduction in CA19–9, continued treatment, and had TTP > 5 months. One patient with colorectal cancer (1.5 mg/kg qw) experienced a durable partial remission, continued treatment, and had TTP > 8 months. Conclusions: RAV12 has activity in recurrent adenocarcinoma. Side effects uncommonly have been dose limiting. The trial continues to define dose/schedule to be recommended for phase 2 testing.

No significant financial relationships to disclose.
Inhibition of colon tumor metastasis in an orthotopic nude mouse model with the dual selective Src/Abl kinase inhibitor, AZD0530
Authors: Phillips K. , Parikh N. , Kuwai T. , Green T. , Gallick G. E.
Published: March 2020

14032

Background: Colorectal cancer is the leading cause of cancer deaths in the USA, largely due to metastatic disease in the liver. The specific activity of the non-receptor protein tyrosine kinase, Src, is increased during colon tumor progression and contributes to metastasis; thus, Src inhibitors may have efficacy in the treatment of advanced-stage colon cancer. AZD0530 is a potent, orally available Src/Abl kinase inhibitor currently in clinical trials. Methods: We examined the effects of AZD0530 alone and in combination with oxaliplatin on two colon tumor cell lines, KM12L4 and LS174T in vitro and in orthotopic nude mouse models. AZD0530 inhibits in vitro proliferation and cellular migration of KM12L4 and LS174T, and decreases phosphorylation of the Src substrates FAK and paxillin in a dose- dependent manner. To examine the ability of AZD0530 to affect colon cancer growth and metastasis in a nude mouse model, we inoculated 1x106 cells into the cecum of nude mice. After 2 weeks, mice were treated with vehicle (n=12), AZD0530 alone (n=13; 25mg/kg/day by oral gavage), oxaliplatin alone (n=13; 5mg/kg/day twice weekly), or AZD0530 and oxaliplatin (n=13). Mice were sacrificed after 4 weeks of treatment. Results: Tumor incidence in all groups of mice was greater than 80%. Mice treated with AZD0530 had an average cecal tumor weight of 0.26 g compared with 0.32 g for control (P=0.24). In contrast, the incidence of liver metastases was reduced by 50% in mice treated with AZD0530 alone or oxaliplatin alone, and no mice developed pathologically evident liver metastases when the combination of inhibitors was used. Furthermore, in mice that developed liver metastases, AZD0530 decreased the size of these metastases 9-fold compared with control. Immunohistochemical analysis of primary cecal tumors demonstrated a decrease in phosphorylated Src (Y418) and FAK (Y861) in AZD0530-treated mice compared with control. Conclusions: These results support further investigation of AZD0530, both alone and in combination, as a new therapeutic strategy for the treatment of advanced colon cancer.

No significant financial relationships to disclose.
Effect of bispecific antisense oligos on prostate cancer cell growth
Authors: Rubenstein M. , Tsui P. , Christensen K. A. , Guinan P.
Published: March 2020

14034

Background: Antisense oligos against TGF-a (MR1), EGFR (MR2), and bcl-2 (MR4) are efficacious against prostate tumors. Methods: To enhance activity “bispecifics” were constructed. A pair recognized TGF-a and EGFR mRNA binding sites (TGF-a/EGFR [MR12] and EGFR/TGF-a [MR21]), another pair EGFR and bcl-2 (EGFR/bcl-2 [MR24] and bcl-2/EGFR [MR42]). Pairs differ in 5’ to 3’ binding site orientation, and were tested in vitro against prostate tumor cells. Following cell attachment, incubations were for 2 days with the agents followed by 2 days in their absence. Results: Bispecifics (6.25uM) are at least as effective as monospecifics and EGFR contributed to most first pair activity. Against PC-3 and LNCaP cells, MR2, MR4, MR12, MR21, MR24 and MR42 significantly inhibited growth. PC-3 cells were then incubated ± a chemotherapeutic agent. With LD50 Cytoxan, MR2, MR4, MR24, MR42 significantly inhibited 47.3, 45.7, 68.3 and 64.9%; with LD50 Taxol MR2, MR4, MR24, MR42 significantly inhibited 49.8, 45.8, 64.1 and 59.2%; and with LD50 DES MR2, MR4, MR24, MR42 significantly inhibited 66.6, 67.6, 64.3 and 67.2% respectively. Each agent significantly increased the inhibition produced by either oligo alone. LNCaP cells were also incubated with mono- and bispecific oligos ± chemotherapeutics. MR2, MR4, MR24, MR42 produced significant inhibitions of 57.4, 58.4, 69.4 and 68.6% for Cytoxan; 70.4, 70.1, 73.6 and 74.0% for Taxol; and 49.8, 50.1, 59.6 and 53.9% respectively for DES. A complete PC-3 experiment compared MR1, MR2, MR4, MR12, MR21, MR24 and MR42 in the presence of LD50 Cytoxan. Each oligo combined with Cytoxan significantly inhibited: MR1 by 51.0, MR2 by 55.0, MR4 by 58.0; MR12 by 56.0; MR21 by 61.1, MR24 by 65.5 and MR42 by 66.0%. Bispecifics directed against two different pathways, MR24 and MR42 were the most effective. A complete LNCaP experiment compared the same series of oligos in the presence of LD50 Cytoxan. Each oligo combined with Cytoxan significantly inhibited: MR1 by 49.0, MR2 by 50.0, MR4 by 53.0; MR12 by 52.0; MR21 by 58.6, MR24 by 53.9 and MR42 by 58.0%. Conclusions: Bispecific oligos significantly advance antisense technology and could play a role treating prostate cancer, particularly when combined with chemotherapy. Bispecifics against proteins associated with different growth pathways may be best.

No significant financial relationships to disclose.
Preliminary results from a phase I/II study of perillyl alcohol intranasal administration in adults with recurrent malignant gliomas
Authors: Da Fonseca C. O. , Schwartsmann Gilberto
Published: March 2020

14037

Background: Deregulated p21-Ras function, as a result of mutation, overexpresion or growth factor-induced overactivation, contributes to growth of malignant gliomas. Perillyl alcohol (POH) is a monoterpene with preclinical antitumor activity in several types of tumor in rodent models and is currently under phase I and phase II clinical trials. Its proposed mechanism of action involves inhibition of post- translational isoprenylation of small G proteins, including p21-Ras, thereby blocking signal transduction Methods: The intranasal delivery is a practical and non-invasive approach that allows therapeutic agents, which do not cross the blood brain barrier (BBB) to enter the Central Nervous System (CNS), reducing unwanted systemic side effects. Applying this method we performed a phase I / II study with POH intranasal administration in patients with relapsed malignant gliomas after standard treatment: surgery, radiotherapy and chemotherapy. POH was administrated in concentration 0.3% volume/volume (55mg) 4 times daily. Results: Thirty-seven patients enrolled being 29 with glioblastoma multiform (GBM), 5 with grade III astrocytoma (AA) and 3 with anaplastic oligodendroglioma (AO). The results showed that POH is well tolerated and display at 6 months of treatment partial responses in 1 patient (3,4%) with GBM and 1 patient (33,3%) with AO; stable disease in 13 patients (44,8%) with GBM, 3 patients (60%) with AA and 1 patient (33,3%) with AO; progressive response in 15 patient (51,7%) with GBM, 2 patients (40%) with AA e 1 patient (33,3%) with AO. Conclusions: The POH intranasal delivery in a concentration of 0.3% volume/volume (55 mg) is well tolerated, and may prolong survival in patients with relapsed malignant gliomas patients.

No significant financial relationships to disclose.
Proteasome inhibition with bortezomib (BORT) in combination with topotecan : A phase I solid tumor trial
Authors: Chen G. Q. , Vikstrom B. , Beckett L. , Scudder S. , Lara P. N. , Lau D. , Gandara D. , Davies A.
Published: March 2020

14047

Background: Topoisomerase-I (Topo-I) down-regulation via the ubiquitin/26S proteasome pathway has been associated with drug resistance. We hypothesize that proteasome inhibition by BORT would enhance topotecan sensitivity by preventing down-regulation of topo-I. This phase I trial examined the safety and feasibility of combining weekly BORT with topotecan in patients (pts) with advanced solid tumors. Methods: Pts on 1 of 4 dose levels were treated with topotecan (3–4 mg/m2) IV and BORT (1–1.6 mg/m2) IV on days 1, 8, and 15. BORT was given 3 hours after topotecan based on preclinical studies (Flannery, 2003). Cycles were repeated q28 days. After 6 cycles, pts without progression could remain on BORT alone. Dose limiting toxicity (DLT) was defined as: grade (Gr) 4 platelets (plts) (<25K), Gr3 plts (<50K) with bleeding, febrile neutropenia, neutropenia with infection, Gr3 neuropathy, or any=Gr3 non-heme toxicity. Results: 15 pts were accrued. Pt characteristics: Median age 58 years (range 25–73); Male sex:7; Performance status =80%:13. Two DLTs were seen at dose level 4 (Topotecan 4 mg/m2 and BORT 1.6mg/m2): Gr 4 plts (1) and Gr 3 neutropenia with infection (1). Few other toxicities > Gr3 were seen past cycle 1. Of 11 evaluable pts, 1 had a partial response (small cell bladder). 4 had stable disease (lung, brain, adenoid cystic and prostate). Conclusions: Weekly BORT and topotecan is feasible and well tolerated. The recommended phase II dose is topotecan 3mg/m2 and BORT 1.6mg/m2 IV weekly (3 of 4). An expanded cohort at this dose will examine efficacy of this combination in pts with small cell cancers. Correlative studies evaluating serum markers for proteasome inhibition and topo-1 activity are ongoing.

No significant financial relationships to disclose.
Different responses of circulating endothelial progenitor cells and VEGF-plasma concentrations to low-dose metronomic and conventional chemotherapy
Authors: Trefzer T. , Stoelting S. , Lemke A. , Kisro J. , Steinke A. , Peters S. O. , Wagner T.
Published: March 2020

14053

Background: Endothelial progenitor cells (EPCs) may participate in tumor angiogenesis by providing cellular supply. This is a study that compares the effects of two conventional combination chemotherapy schedules to low-dose metronomic trofosfamide (an oral derivate of cyclophosphamide), using the number of circulating EPCs and vascular endothelial growth factor (VEGF) plasma levels in cancer patients. Methods: We measured circulating EPC and VEGF levels in 24 patients that received conventional chemotherapy for either breast cancer in an adjuvant setting or malignant lymphoma, and in 18 patients receiving metronomic chemotherapy with or without celecoxib for advanced cancer. Blood samples were obtained three times: before starting chemotherapy, 10 and 21 days after starting chemotherapy. Peripheral blood EPC levels were determined by fluorescence flow cytometry and defined by CD34- and VEGF-R2-positivity. VEGF plasma concentration was determined by ELISA. Results: The number of circulating EPCs showed a two-fold increase 21 days after conventional chemotherapy but a significant decrease under metronomic chemotherapy. VEGF-plasma-concentrations remained stable in patients under metronomic chemotherapy but significantly increased under conventional chemotherapy.This increase in VEGF plasma levels occurred even in patients that received chemotherapy in an adjuvant setting in supposed absence of tumor. Conclusions: Low-dose metronomic trofosfamide significantly decreased circulating EPCs while conventional chemotherapy increased both the number of circulating EPC and VEGF-concentrations. The increase of VEGF even in an adjuvant setting of chemotherapy without the presence of a tumor may be caused by chemotherapy-induced endothelial cell destruction. Metronomic scheduling of certain cytotoxic drugs may thus prevent tumor progression by inhibiting both tumor cell proliferation and angiogenesis.

No significant financial relationships to disclose.
Phase I study of pemetrexed (P) plus gemcitabine (G) in advanced solid tumors (ST)
Authors: Fury M. G. , Larkin J. , Gerst S. R. , Sabbatini P. , Konner J. , Orlando M. , Tai D. F. , Goss T. , Aghajanian C. , Hensley M. L.
Published: March 2020

14055

Background: P is active in multiple ST types, and preclinical data support P synergy with G. Methods: Eligible advanced ST patients (pts) with no prior P or G, no prior radiotherapy (RT) to ≡ 25% of the marrow, Karnofsky Performance Status ≡ 70%, and adequate organ function enrolled in cohorts (C) of 3, expanding to 6 if dose-limiting toxicity (DLT) occurred. P was given at 300 (C1), 400 (C2), 500 (C3) or 600 (C4) mg/m2 followed by G at 1500 mg/m2 q 14 days (d) without granulocyte-colony stimulating factor. Vitamin B12 and folate supplementation were given. Response was assessed by RECIST Results: 29 pts (median number prior regimens 2, range 1–5; 66% with prior RT) enrolled and are evaluable for safety; 23 are evaluable for response. There were no DLTs in C1. One pt in C2 was replaced after 1 cycle for progression of disease (PD). Among the next 6 pts, 2 had DLTs (1 G3 thrombocytopenia [TP] treatment delay; 1 neutropenic fever [NF]). C2-R (C2, Revised) re-opened after amendment permitting ≡ 2 prior cytotoxic regimens, no history of brain metastases/brain RT. C2-R enrolled 8 pts with 1 DLT (G3 TP with treatment delay) and 2 pts replaced (1 early PD, 1 no documented duration of neutropenia [NP]). C3 had 0/3 pts with DLT. C4 had 2/3 pts with DLTs (1 G4 hyponatremia; 1 herpes zoster-related treatment delay). C3 has been expanded to 5 of 6 planned patients, one with DLT (NF, G4 TP). Toxicities per cycle (n= 189 cycles, 29 patients): include NP-G3 (23%), G4 (14%); TP-G3 (2%); WBC-G3 (30%), G4 (4%); lymphopenia-G3 (11%), Hgb-G3 (4%); G3-NF (1%). 3/23 (13%) had objective partial responses (2 head and neck squamous cell cancer, HNSCC; 1 nasopharyngeal cancer, NPC), 4 stable disease (SD), 16 PD. (1 pt, no measurable disease at baseline; 5 pts, too early for response assesment). Conclusions: G + P is well-tolerated, and yields objective responses in HNSCC and NPC. C3 (P 500 mg/m2 + G 1500 mg/m2 q 14 d) was the phase II recommended dose in another phase I study of this regimen (Melemed ASCO 2005). Our final results will be available for ASCO 2007.

No significant financial relationships to disclose.
Phase 1, pharmacokinetic (PK) and pharmacodynamic (PD) study of oral alvespimycin (A; KOS-1022; 17-DMAG): Two different schedules in patients with advanced malignancies
Authors: Flaherty K. T. , Gore L. , Avadhani A. , Leong S. , Harlacker K. , Zhong Z. , Johnson R. G. , Hannah A. L. , O’Dwyer P. , Eckhardt S. G.
Published: March 2020

14059

Background: A is ∼3–5 fold more potent compared to 17-AAG (the first Hsp90 inhibitor to enter clinical testing), based on in vitro cytotoxicity or the MTD in toxicology studies; it is water-soluble; and oral bioavailability in dogs is estimated at 40%. Toxicity in the dog included kidney, intestinal and liver findings. This study was conducted to determine the toxicity, MTD, recommended phase 2 dose (RP2D), PK and PD of A in pts with solid tumors. Methods: Escalating doses of A were given PO on 2 different schedules: every other day or daily for 4 out of 6 weeks. An initial IV dose was given to calculate absolute bioavailability. PK was evaluated after the IV dose, Day 1 and 21 of oral dosing. PBLs were collected to investigate changes in intracellular signaling proteins by immunoblot (Days 1 and 21 at 1, 3, 24 and 48 hours post-dose). Results: 28 pts were enrolled: 24 on the QOD schedule at doses of 5 (n=4), 10 (n=4), 20 (n=8), 30 (n=5) and 40 mg (n=3); 4 pts received 10mg on the QD schedule. 50% were male, median age/ECOG PS 55 and 0; median prior regimen 3. DLT has not yet been observed. Common drug-related toxicities (n=23): fatigue 43%, nausea 24%, anorexia 19%, proteinuria 19%, and peripheral edema 14%. Of these, fatigue and peripheral edema appear to be possibly dose-related. Drug-related Grade 3–4 toxicity (one patient each) included anemia, neutropenia, peripheral edema, hypokalemia, pain in extremity and hypoxia. For pts with full PK data (n=14), bioavailability equaled 51% and 49% on Day 1 and 21, and was not dose-dependent. Mean Day 21 AUCinf for the 5 to 30 mg/m2 levels equaled 91, 166, 542 and 1889 ng*h/mL. One pt with 3-fold increase in AUCinf comparing Day 1 and 21 dose had been started on dronabinol, a CYP2C9 inhibitor. One pt with fibrosarcoma (4 prior regimens) had necrotic changes in the tumor in the axilla with improved symptoms (active at 5+ months). Additional pts with SD include hemangioendothelioma (7 months), melanoma (6+ months), and renal cell (5 months). Induction in Hsp70 at the 30 mg dose level was seen pre-dose on Day 21 with maximal induction at 24 hours post-dose. Conclusions: Dose escalation continues on both schedules in order to define a RP2D. Toxicity is acceptable. Early signs of activity have been observed.

[Table: see text]
Sensitivity of primary hematopoietic tumors and tumor lines to the novel HDAC inhibitor PCI-24781
Authors: Balasubramanian S. , Ramos J. , Sirisawad M. , Buggy J. J. , Miller R. A. , Phan S.
Published: March 2020

14062

PCI-24781 (formerly CRA-024781) is a novel HDAC inhibitor that is in phase I clinical trials in patients with solid and hematopoietic malignancies. PCI-24781 has favorable pharmacokinetic and pharmacodynamic profiles in animal models and in humans. In the present study we show that PCI-24781 potently induces cell death in a variety of hematopoietic cell lines derived from B-cell, T-cell and myeloid malignancies, and in primary acute leukemic blasts from bone marrow aspirates. In tumor cell lines, growth inhibition and apoptosis were noted at drug concentrations ≡ 0.125 μM and were accompanied by known biochemical markers of HDAC inhibition including histone and tubulin hyperacetylation. To demonstrate the potential clinical utility of PCI-24781 in hematologic tumors, primary leukemia samples were isolated from patients and screened for resistance to PCI-24781-induced growth arrest in vitro. Of these 25 primary samples (10 acute myelogenous leukemia (AML), 6 multiple myeloma (MM) and 9 acute lymphocytic leukemia (ALL)), some of which were derived from patients who had failed standard therapy, none was resistant to PCI-24781 at 0.5 μM and only 4 (1 AML, 2 MM, and 1 ALL) were considered resistant at 50 nM. Gene expression analysis using DNA microarrays on these primary tumor samples revealed alterations of gene expression consistent with HDAC inhibition and defined potential pathways of activity for this compound in these tumors. These results demonstrate that hematopoietic tumors and tumor-derived cell lines are highly sensitive in vitro to the novel HDAC inhibitor PCI-24781. The high sensitivity of primary tumor cells to treatment with PCI-24781 in vitro coupled with the favorable pharmacokinetics of this compound in humans suggests that patients with hematopoietic malignancies would be responsive to treatment with PCI-24781 in clinical trials.

No significant financial relationships to disclose.
Early prediction of anti-epidermal growth factor receptor (EGFR) therapy with 18[F]FDG-PET: A preclinical and clinical correlation
Authors: Ma W. , Jimeno A. , Kulesza P. , Chan A. , Zhang X. , Messersmith W. A. , Gillison M. L. , Pomper M. G. , Forastiere A. A. , Hidalgo M.
Published: March 2020

14064

Background: Predictive biomarkers for response to anti-EGFR therapy are of significant clinical importance. We hypothesized that early changes in 18[F]FDG tumor uptake predict response to anti-EGFR therapy. We investigated this in a mice model and correlated these findings with human patients undergoing anti-EGFR therapy. Methods: Mice bearing two head and neck squamous cell cancer (HnNSCC) xenografts (Hep2, Cal27) received vehicle or erlotinib for 3 weeks. 18[F]FDG uptake were imaged with small animal micro- PET at baseline and after 1 week of therapy, and reported in SUVmax. For clinical correlation, human patients with HnNSCC receiving erlotinib were identified from an ongoing clinical trial. 18[F]FDG-PET images were obtained at baseline and after 2 weeks of erlotinib monotherapy. Tumor specimens were obtained by fine-needle aspiration biopsy at the same time as PET. A panel of pharmacodynamic markers (including Ki-67 and pMAPK) were assessed . Results: Hep2 was resistant to erlotinib therapy (Tumor/Control [T/C]: 0.8) while Cal27 was sensitive (T/C: 0.2). SUVmax in the resistant Hep2 xenografts was not significantly different from the control (86% ± 70% of control) while SUVmax in the treated sensitive Cal27 xenografts showed a significant decrease than control (-2% ± 7%). Ki-67 in the treated Hep2 was not significantly different from control while that in Cal27 was <50% of control. For clinical correlation, ki-67 score was higher after 2 weeks in the patient resistant to erlotinib monotherapy by 18[F]FDG-PET (SUVmax increased by 14%, ± 11%) and was lower in the patient who was 18[F]FDG-PET sensitive (SUVmax decreased by 56%, ± 13%). pMAPK decreased in all cases and had a poor correlation with efficacy. Conclusions: Early dynamic changes in 18[F]FDG tumor uptake is predictive of response to anti-EGFR therapy and correlates with changes in ki-67 expression, both in a preclinical and a clinical scenario.

No significant financial relationships to disclose.
Technical criteria of “clinical readiness” to qualify validated pharmacodynamic (PD) assays for clinical trial use
Authors: Parchment R. E. , Kinders R. J. , Hollingshead M. , Dancey J. E. , Tomaszewski J. E. , Doroshow J. H.
Published: March 2020

14068

Background: Pharmacodynamics may inform drug development decisions by providing evidence of drug action on molecular target and additional data supporting dose/schedule. While a bad decision when selecting the most appropriate PD assay to use is not fatal to completing Phase 1–2 trials, assays that cannot produce meaningful scientific results create an ethical dilemma if patients are being subjected to invasive procedures, and weaken decision making. In some exploratory IND trials, the primary endpoint is demonstrating a PD effect of investigational drug on molecular target, so selection of a PD assay becomes a critical decision. Methods: SAIC-Frederick’s applied research program developed, validated and implemented a PD assay to quantify drug-induced PARP inhibition as the primary endpoint of a Phase 0 trial of ABT-888 at the NIH Clinical Center (see Kinders et al, Kummar et al). This project has been analyzed to identify technical requirements critical for establishing the clinical readiness of a PD assay. Results: Many more technical requirements arose than were originally expected, but the requirements fell into three conceptual categories: assay validation, clinical feasibility and therapeutic relevance. Assay validation requires formal assessment of analytical performance on a set of standards and specimens relevant to the trial, including storage stability of the analyte. Clinical feasibility requires the PD assay to yield analytically valid results in preclinical models when using clinically relevant tissue acquisition, handling procedures and biopsy scheduling, including significant drug effect in needle biopsies when confounded by stress, previous biopsy, biopsy method, and co-medications. Therapeutic relevance requires a relationship between dose/exposure, target modulation quantified with the PD assay, and a disease outcome of interest. Conclusions: “Clinical readiness” sets a very high standard for accepting PD assays into clinical trials, and a highly integrated team is required to meet it. However, the use of such rigorous assay development criteria holds much promise for the clinical development of molecularly targeted agents. Funded by NCI Contract N01-CO-12400.

No significant financial relationships to disclose.
Activity of the aurora kinase inhibitor MK-0457 (VX-680) in combination with taxotere
Authors: Hoover R. R. , Harding M. W.
Published: March 2020

14069

Background: MK-0457 (VX-680) is a reversible kinase inhibitor that targets Aurora A, B and C (Ki’s of 0.7, 18 and 4.6 nM, respectively). Clinical trials are on-going in patients with solid tumors and hematologic malignancies. This study evaluated the in vitro combination of MK-0457 and Taxotere (TXT) to determine if inhibition of Aurora kinases, which are critical for cytokinesis and spindle assembly may synergize with a microtubule targeted agent. Methods: A panel of cancer cell lines (that were either wild-type, mutant, or null for p53) and a matched pair of A549 cell lines either expressing p53 shRNA or vector control (A549p53- and A549p53+) were evaluated for sensitivity to MK-0457 and/or TXT. Viability was measured by Cell Titer Glo (Promega). Cell cycle profiles were determined by FACS analysis, and colony formation (CFU) assays were performed in soft agar and on plastic. Combination effects were evaluated by the Bliss Independence method. Results: Single agent IC50’s ranged from 50 nM to > 5 μM for MK-0457 and 0.4 to 10 nM for TXT. FACS analysis of DNA content showed MK-0457 induced G2/M arrest and polyploidy, characteristic of Aurora kinase inhibition. The effects of MK-0457 and TXT combination ranged from antagonism to synergy and were concentration, cell context, and regimen dependent. In 3–4 day viability assays simultaneous combination of MK-0457 + TXT did not show synergy, however sequential exposure showed synergy at low concentrations of both agents. In contrast to the short-term viability assays, simultaneous exposure to MK-0457 + TXT in long term survival assays (i.e. CFU on plastic or soft agar) resulted in enhanced cell death compared to the single agents. Conclusions: The synergy observed for MK-0457 + TXT, especially in long term survival assays, supported the evaluation of this combination in xenograft models (described in B. Furey et al., ASCO 2006).

No significant financial relationships to disclose.
A phase I dose finding study of E7080 in patients (pts) with advanced malignancies
Authors: Glen H. , Boss D. , Evans T. R. , Roelvink M. , Saro J. M. , Bezodis P. , Copalu W. , Das A. , Crosswell G. , Schellens J. H.
Published: March 2020

14073

Background: E7080 is a potent inhibitor of the split-kinase family of transmembrane growth factor receptors including Flt-1 and KDR. In addition, E7080 also potently inhibits FGFR1 and PDGFRβ tyrosine kinase activities. In an in vitro angiogenic model, E7080 inhibited VEGF-driven umbilical vein endothelial cell (HUVEC) proliferation and tube formation. Using H460 and Colo205 mouse xenograft models, E7080 significantly inhibited tumor growth at doses from 1 to 100 mg/kg. Methods: A Phase I and pharmacologic study was conducted to determine the Maximum Tolerated Dose (MTD) and Dose Limiting Toxicity (DLT) of E7080 in pts with advanced malignancies. Eligible pts with adequate hematologic, renal and hepatic function, and with tumors refractory to standard therapies, were treated with E7080 administered orally on a once daily continuous dose schedule in cycles of 4 weeks. Dose escalation was performed according to an accelerated design of 100% dose increases in subsequent cohorts until the first patient experienced ≥ CTC grade 2 toxicity. Results: As of November 17th, 2006, 37 pts with documented progressive disease (PD) have been treated at the following dose levels: 0.2 mg/d, 0.4 mg/d, 0.8 mg/d, 1.6 mg/d, 3.2 mg/d, 6.4 mg/d, 12.5 mg/d and 16 mg/d. Median number of cycles was 1 (0–11). Median age = 60 years (25–84), 20 pts were male, 17 were female. 37 pts were evaluable for toxicity. 1 patient experienced grade 3 hypertension (DLT) in the 16 mg/d cohort, other related toxicities ≥ grade 2 include hemorrhage and thrombosis (n=1), tachycardia (n=1), febrile neutropenia (n=1), proteinuria (n=1), thrombocytopenia (n=1) and hypertension (n=1). MTD has not been reached and this study is continuing to recruit. 24 pts were evaluable for efficacy, 3 are too early to evaluate. 1 patient had a partial response (high grade sarcoma) after 2 cycles, and 15 pts experienced stable disease from 2+ to 11 months, and 11 pts continue on therapy. Pharmacokinetic (PK) studies: preliminary data suggest that the PK is linear over the range 0.2 mg/d to 12.5 mg/d. Conclusions: E7080 is safe and well tolerated at doses acheived so far and has shown promising first signs of anti-cancer activity associated with potent activity in pre-clinical models. Clinical and PK data from subsequent doses and follow- up will be presented.

No significant financial relationships to disclose.
Relation between lactate dehydrogenase (LDH) during two first cycles in patients with metastatic renal cell carcinoma (mRCC) treated with sunitinib (SU)
Authors: Medioni J. , Arakelian N. , Fournier L. , Helley D. , Thiam R. , Banu E. , Cuenod C. , Oudard S.
Published: March 2020

14079

Background: LDH is a biomarker of cellular turn over. We evaluated LDH as a surrogate marker of tumor response and tumor necrosis during first cycles of SU. Methods: This is a single center study of immunorefractory patients with mRCC treated in second line with SU (50 mg, 4 weeks on, 2 weeks off). All patients performed CT scan and had serially measured tumor necrosis. Tumor response using the RECIST criteria was evaluated at the end of cycle 2 (C2). Tumor necrosis index was calculated at baseline, at the end of cycle 1 (C1) and C2 by measuring the percent area of the tumor with no enhancement after injection using an electronic calliper. LDH was measured at baseline (D1) and at the end (D28) of C1 and C2 using LD 200 kit on Synchron LX20 (Beckman Coulter). Relations between LDH and tumor response were studied using multivariate analysis of variance (MANOVA) with repeated measures and between LDH and tumor necrosis using Pearson correlation test. Results: Between Feb 2005 and Aug 2006, 56 patients were analyzed (75% men,), 91% Clear Cell Carcinoma. Mean age was 59 years (range: 30–81) and 62% has a performance status ECOG of 0. Median number of metastatic sites was 2 and mean hemoglobin level was 12.7 gr/dl. After C2, 12 (21%) partial responses, 35 (63%) stable diseases and 9 (16%) progressive diseases were noted. Results of LDH are summarized in the table . Mean LDH was statistically different between along time (p < 0.0001). LDH levels were statistically different between responders-stables and progressors patients (p = 0.004). There is a trend for a statistical relation between the index of necrosis and the tumor response measured by RECIST criteria (p = 0.08). No correlation was found between LDH and tumor necrosis. Conclusions: LDH level is higher in progressors compared to responders-stables patients. LDH level is not correlated to tumor necrosis measured by CT scan.

[Table: see text]

No significant financial relationships to disclose.
CD40 ligand gene therapy directly induces apoptosis in carcinomas
Authors: Palmer D. H. , El-Metwali T. , Milner A. E. , Hodgkin E. , Young L. S.
Published: March 2020

14082

Background: CD40 is a member of the TNFReceptor superfamily expressed on a variety of immune cells. Interaction of CD40 with its ligand (CD40L) plays a key role in orchestrating immune responses. We have shown that CD40 is also expressed on a variety of carcinomas. CD40 ligation in this context can induce cancer cell death. Thus CD40 represents a rational target with direct cytotoxicity and induction of anti-tumour immunity. Methods: We generated an adenovirus encoding CD40L (AdCD40L) and investigated its effect in CD40-expressing carcinoma cells (bladder, ovary, hepatocellular) as follows:(i)comparison of Ad-delivered CD40L with recombinant soluble ligand (rsCD40L);(ii)inhibition of cleavage of CD40L from the cell membrane using metalloproteinase inhibitors (MMPI);(iii)generation of a mutant CD40L resistant to MMP cleavage. Results: (i)CD40 ligation in carcinomas stimulates survival and apoptotic signaling. rsCD40L is only cytotoxic in the presence of protein synthesis inhibition, which blocks survival pathways and allows apoptosis. In contrast, AdCD40L is cytotoxic even in the absence of protein synthesis inhibitor;(ii)AdCD40L results in expression at the cell membrane but this is cleaved from the membrane and detected as soluble ligand in supernatant of infected cells. Cleavage is inhibited by MMPI and this increases apoptosis, suggesting that membrane-bound CD40L is a more potent apoptotic stimulus than soluble ligand;(iii)we identified the cleavage site of CD40L and generated a mutant by deletion of this region and cloned this mCD40L into an adenovirus. AdmCD40L results in membrane expression of CD40L, with no detectable soluble CD40L, confirming its resistance to cleavage. AdmCD40L is significantly more cytotoxic than wild-type AdCD40L. Conclusions: The effect of CD40 ligation depends on the cellular context and the method of CD40L delivery: adenovirus delivery results in membrane-bound expression and this provides a stronger apoptotic stimulus than rsCD40L. Apoptosis is further enhanced by a mutant CD40L that is resistant to cleavage from the cell membrane. Thus AdmCD40L is a novel therapeutic that is a potent inducer of apoptosis. Studies are underway to assess these effects in vivo and to elucidate the contribution of immunostimulatory effects in syngeneic models.

No significant financial relationships to disclose.
Measurement of gene amplification in FFPE tumor tissues by quantitative real-time PCR in comparison with FISH
Authors: Boyer S. , Yang J.
Published: March 2020

14086

Background: Amplified oncogenes such as HER2 and EGFR have proven to be good targets for molecular therapy. Recent retrospective analyses suggest that EGFR amplification predicate better responses to EGFR kinase inhibitors. However, different methods were used to measure EGFR gene amplification and polysomy in these studies, including FISH and various real-time quantitative PCR platforms, which may have led to the different conclusions reported by these groups. In this study, we compared Taqman- QPCR with FISH in scoring EGFR and HER2 amplification and EGFR polysomy on 42 FFPE tumor samples. Methods: DNA was prepared from 42 FFPE tumor tissues of breast, colon, lung, and stomach origin. Taqman QPCR was carried out according to manufacturer’s protocol on an ABI-7900HT instrument. FISH analysis for EGFR and HER2 amplification was carried out according to Vysis standard protocols. Results: QPCR reference probes for measuring EGFR and HER2 amplification were designed against DNA fragments near the centromeric regions of chromosome 7 and chromosome 17, respectively. A QPCR probe for RNase P on chromosome 6 was designed and used as a reference for measuring EGFR polysomy. HER2 amplification was detected in ∼25% FFPE breast tumor samples by both FISH and QPCR methods, although amplification scores were consistently higher by QPCR than by FISH. EGFR amplification was rare in all samples screened but high polysomy (6 copy and above) was detected in 40% FFPE gastric tumor samples. Again there was high concordance between FISH and Taqman QPCR results. Conclusions: Our data showed high concordance between Taqman QPCR and FISH in scoring EGFR and HER2 amplification. Taqman QPCR is a highly sensitive, accurate, convenient, and economical technology for measuring EGFR and HER2 amplification and EGFR polysomy in FFPE tissues. With well validated target and reference primers/probes, Taqman QPCR provides an attractive alternative to FISH for measuring oncogene amplification for retrospective studies on archived FFPE samples and for screening patients for clinical trials.

No significant financial relationships to disclose.
Hepatic expression of UGT enzymes and implications in pharmacokinetic variations in Asians during chemotherapy
Authors: Chowbay B. , Lal S. , Shu X. , Jada S. , Ooi L. , Liu Y. , Ong Y.
Published: March 2020

14089

Background: The objective of this study was to screen the promoter and relevant coding regions in the UGT1A1 and UGT1A9 genes and genotypically quantitate expression of these genes in hepatic tissues obtained from Asian cancer patients undergoing hepatectomy. Methods: Non-cancerous Asian hepatic tissues (Chinese: N=36; Malay: N=1; Indian: N=3) were obtained from the institution’s central tissue repository bank. Messenger RNA was isolated from each tissue sample and reverse transcribed to cDNA followed by real time PCR experiments. Results from these gene expression experiments were normalized against GAPDH transcripts. DNA was also extracted from the hepatocytes, screened for variations in the promoter regions (UGT1A1*60, g.3140G>A and UGT1A1*28) and exon 1 (UGT1A1*6) of the UGT1A1 and UGT1A9 (UGT1A9*22, I399C>T) genes. Results: Large interindividual variations were observed in the hepatic expression pattern of UGT1A1 and UGT1A9 mRNA with respect to the different polymorphic variants. There was no significant association between hepatic expression of UGT1A1 mRNA and UGT1A1*60, g-3140G>A and *6 genotypes (P>0.05 in each case). The median expression level of UGT1A1 mRNA was approximately 2-fold higher in liver tissues harboring UGT1A1 6/6 (reference) genotype (N=27, median = 0.478; range: 0.065 to 3.125) compared to the heterozygous 6/7 genotype (N=9, median = 0.254; range: 0.042 to 1.894; P=0.77). The median expression levels of UGT1A9 mRNA were approximately 3-fold higher in liver tissues harboring I399CC genotype (N=3, median = 2.735; range: 0.702 to 3.375) when compared with the homozygous variant TT genotype (N=8, median = 0.973; range: 0.652 to 4.155; P=0.88). Conclusions: The results showed that polymorphisms in the UGT1A genes may influence the expression pattern of the respective proteins and could contribute to the high degree of interpatient variability in the glucuronidation dependent metabolism and elimination of chemotherapeutic agents in Asian cancer patients.

No significant financial relationships to disclose.
Pediatric phase I trial and pharmacokinetic (PK) study of dasatinib: A report from the Children’s Oncology Group
Authors: Aplenc R. , Strauss L. C. , Shusterman S. , Ingle A. M. , Luo R. , Wright J. , Blaney S. , Adamson P. C.
Published: March 2020

14094

Background: Dasatinib is an orally-available tyrosine kinase inhibitor with low-nanomolar activity against SRC-family kinases, BCR-ABL, c-KIT, EPHA2, and the PDGFβ receptor. Methods: A phase I study of dasatinib administered as a single agent in pediatric patients with refractory solid tumors or imatinib refractory Ph+ leukemias is being performed. Dasatinib dose levels of 50, 65, 85 and 110 mg/m2/dose, administered orally twice daily for 28 consecutive days, are being studied. Courses repeat without interruption. Results: 8 pts with solid tumor and 3 pts with Ph+ leukemia (median age 11 yrs, range 7–17) have been enrolled, of whom 7 are fully evaluable for toxicity. At the 50 and 65 mg/m2 dose levels, 0/3 and 0/1 pts experienced DLT. 1 of 3 pts with CML treated at the 50 mg/m2 dose level had Gr 4 hypokalemia. In 4 pts studied at the 50 mg/m2 dose level, the median (range) apparent dasatinib clearance was 238.5 L/hr (220.7 - 332.7), terminal half-life was 2.3 hrs (2.2 - 3.1), and the median Cmax was 55.5 ng/ml (36.0 - 161.1). Conclusions: Dasatinib is well tolerated at a dose of 50 mg/m2 BID on a continuous 28 day dose schedule in pediatric patients. Preliminary PK analysis suggests that drug disposition is similar to that observed in adults. Dose escalation and PK studies are continuing.

No significant financial relationships to disclose.
Dual AKT inhibition with perifosine and GCP enhances apoptosis in prostate cancer: Clinical implications
Authors: Vinall R. L. , Hwa K. , Pan C. , Ghosh P. , Lara P. N. , deVere White R. W.
Published: March 2020

14096

Background: Genistein combined polysaccharide (GCP) has been shown to exhibit anti-cancer properties in both experimental and clinical models of prostate cancer (CaP). Perifosine is an alkylphospholipid with clinical anti-neoplastic activity. Both agents inhibit the AKT signaling pathway. Methods: Prostate cancer cell lines (LNCaP, LNCaP stably transfected with R273H or P151S p53 mutant allele, cds1, and PC3) were treated with GCP, perifosine, or both. IC50 values were established using the MTT assay. Apoptosis was assessed by flow cytometry, Western blot of PARP cleavage, and caspase activity. Clonogenic potential was assessed by colony assay. Status of AKT, p53, p21, AR and PSA was determined by Western blot. All experiments were performed in triplicate. Results: Perifosine inhibited AKT activity in all the cell lines. GCP had little or no effect on AKT activity but reduced AR and PSA levels. The combination of GCP and perifosine further increased the level of AKT inhibition and maintained inhibition for longer when compared to treatment with perifosine alone. Flow cytometric analysis of LNCaP revealed that combination treatment dramatically increased SubG1 levels (23-fold increase versus a 4.4-fold increase for GCP alone and a 6.5% increase for perifosine alone). Apoptosis was confirmed by PARP and caspase analysis. As single agents, the main effect of GCP or perifosine was to induce growth arrest as shown by a decrease in S-phase and increased p21. The cds1 cell line responded similarly to LNCaP, however, cell lines that expressed mutant p53 or were p53 null were not susceptible to GCP/perifosine-induced apoptosis. Combination treatment further decreased the clonogenic potential in all of the cell lines assessed when compared to treatment with the single agents alone. Conclusions: Treatment with a combination of the AKT inhibitors GCP and perifosine dramatically increases apoptosis and/or inhibits clonogenic potential in several CaP cell lines. The effect of the combination treatment on apoptosis appears to be dependent on p53 status. Clinical validation of these findings is warranted. A clinical trial of hormone therapy with or without GCP/perifosine is presently in development.

No significant financial relationships to disclose.
Recruitment experience in a phase 0 trial of ABT-888, an inhibitor of poly (ADP-ribose) polymerase (PARP), in patients (pts) with advanced malignancies
Authors: Gutierrez M. , Kummar S. , Horneffer Y. , Juwara L. , Chen A. , Melillo G. , Pickeral O. , Tomaszewski J. E. , Murgo A. J. , Doroshow J. H.
Published: March 2020

14111

Background: Phase 0 trials conducted under the FDA Exploratory IND Guidance involve limited exposures to study drug with no expectation of clinical benefit. We present our experience in recruiting pts to a Phase 0 study of ABT-888, the first to be conducted at the NCI. Methods: Objective: To determine the ABT-888 dose that inhibits PARP in PBMCs and tumor after a single oral administration. Tumor biopsies at baseline and 3–6 hrs post drug were planned once PARP activity inhibition in PBMCs or target plasma Cmax was achieved. Protocol was approved by the NCI scientific committee, IRB, and CTEP, and was reviewed by the NIH Ethics Committee. Informed consent included an explicit statement acknowledging the non-therapeutic, non-personal benefit nature of the study. Results: Since 6/06, 17 pts screened; 6 enrolled. Reasons for declining participation: Non-therapeutic study (2); biopsy requirement (2); recommendation by oncologist (2) or family member (2); condition requiring treatment (3). Reasons for participation: Altruism (4); altruism + waiting for another study (2). Of 6 pts enrolled, colon cancer (2), lymphoma (2), carcinoid (1), lung cancer (1). Median age: Participants (P) 57 (49–64) and non-participants (NP): 62 (42–78). Median prior therapies: P 3.6 (0–8) and NP: 3 (0–7). Referral origin for P: Prior participation on NCI studies (2), prior pt-NCI physician relationship (4). Ethnicity of P: White (5) and Hispanic (1). 3 of 6 pts underwent serial biopsies as required by protocol. Conclusion: Phase 0 recruitment is feasible and the accrual can be completed in a timely fashion, but depends on pt altruism and prior physician-pt relationship. These studies can be ethically conducted provided that interventions are of minimal risk, the study experience is commensurate with pt expectations, and that results are invaluable to subsequent pts.

No significant financial relationships to disclose.
Predicting and monitoring tumor response to epidermal growth factor receptor inhibitor gefitinib in patients with locally advanced esophageal adenocarcinoma
Authors: Gibson M. K. , Mezzadra H. , Kleinberg L. , Jagannath S. , Brock M. , Abdallah N. , Rudek M. , Berman D. , Forastiere A. , Altiok S.
Published: March 2020

14112

Background: This study aimed to validate an ex vivo chemosensitivity assay to measure the pharmacodynamic effect of gefitinib on esophageal adenocarcinoma (EAC) prior to treatment with pre-operative concomitant chemoradiotherapy (CRT). Methods: A 14 day run-in period with 250 mg/day of gefitinib preceded CRT. Endoscopic biopsies (D 0 and 14) in 4 patients with T2–3N0/1M0/1a EAC were analyzed by ex vivo chemosensitivity assay. Day 0 tissue was exposed to gefitinib ex vivo, then tumor was exposed to gefitinib for 14 days in vivo (ie in the patient). Phosphorylation of the EGFR, raf/MEK/ERK and PI3/AKT pathways was measured by Western blot. Profiles were compared for correlation between ex vivo and in vivo exposure, and patterns were correlated with response to CRT. The effects were also characterized by immunohistochemistry (IHC). EGFR, K-Ras, and PI3K mutations, serum concentrations of gefitinib and PTEN status were measured as potential confounders. Results: One patient with stage T3N1 died of unexplained hemorrhage during surgery. Three had clinical and path stages of: T3N1/T0N0, T3N0/T3N0, T3N1/T2N1. Gefitinib levels were constant, confirming exposure of target tissue to the drug. Ex vivo exposure yielded four distinct pathway patterns. The exact same patterns were seen after in vivo exposure. No mutations were identified in exons 18–21 of the EGFR, exons 2/3 of K-ras or exons 9/22 of PI3K. PTEN levels were similar in all tumors. PCNA expression correlated with raf/MEK/ERK pathway inhibition, but not with inhibition of EGFR activity. IHC correlated with Western blot for expression of EGFR, and phospho- and total ERK levels. No correlation was observed between gefitinib effect and pathologic response to CRT. Conclusions: This study used a novel ex vivo chemosensitivity assay to demonstrate the activity of gefitinib to inhibit target in tumor tissue obtained from patients with EAC. The exact correlation of pre- and post-treatment profiles suggests potential use in the pre-treatment setting to predict in vivo effects of targeted therapies. This approach may facilitate the further refinement of patient selection to maximize potential benefit while sparing patients unlikely to respond to a given agent.

No significant financial relationships to disclose.