bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase I dose escalation study of continuous oral dosing of OSI-906, an insulin like growth factor-1 receptor (IGF-1R) tyrosine kinase inhibitor, in patients with advanced solid tumors
Authors: Lindsay C. R. , Chan E. , Evans T. R. , Campbell S. , Bell P. , Stephens A. W. , Franke A. , Poondru S. , Rothenberg M. L. , Puzanov I.
Published: August 2020

2559

Background: OSI-906 is a potent small molecule inhibitor of IGF-1R, a receptor tyrosine kinase activated by insulin like growth factor, which is overexpressed in numerous malignancies and implicated in resistance to chemotherapy. Methods: Patients with advanced cancer entered escalating dose cohorts of OSI-906. Study objectives included assessment of: safety; determination of MTD; pharmacokinetics (PK), pharmacodynamics (PD) including IGF-1R levels in peripheral blood cells; and tumor response (RECIST). Results: To date, 32 pts have been treated (22M/10F, median age 62y) at 10, 20, 40, 75, 150, and 300 mg QD and at 20, 40 mg and 75 mg BID. Median number of weeks on trial was 6 (range 0–44). No DLTs have been observed. Hyperglycemia (5/20 pts) related to OSI-906 was transient and mild (grade 1 only). In addition to hyperglycemia, grade 1–2 nausea and vomiting (5/20 pts) were the most frequent related adverse events (AEs). There was grade 3 elevated lipase in 1 pt. At doses of 10–150 mg, OSI-906 exhibited linear PK, median terminal t1/2 ranged from 2.18–4.30 hr, AUC0-inf from 284–10200 ng.hr/mL, and Cmax 76.6–1440 ng/mL. There was no relationship between glucose or insulin levels and OSI-906 plasma concentrations. Stable disease > 12 weeks was seen in 7/20 pts (range 12–34 weeks), including 1 pt each with thymic (27 w), adrenocortical (28w), and colorectal (34w) cancer. PD data on IGF-1R phosphorylation will be presented. Conclusions: Plasma concentrations of OSI-906 achieved in this trial exceed concentrations required for antitumor efficacy in preclinical models. PD target modulation and preliminary anti-tumor activity have been observed. It is interesting to note that clinically meaningful hyperglycemia has yet to occur. Minimal toxicity was observed and further dose escalation is in progress.

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Pharmacodynamic effects of MLN4924, a novel NAE inhibitor, on blood and skin
Authors: Kauh J. S. , Cohen R. , Clarke J. W. , Harvey R. , Mulligan G. , Venkatakrishnan K. , Berger A. , Smith P. G. , Petruzelli L. , Sarantopoulos J.
Published: August 2020

2562

Background: MLN4924 is a first-in-class, small molecule inhibitor of NEDD8 Activating Enzyme (NAE) with potent antitumor activity in animal models. Inhibition of NAE by MLN4924 leads to decreased neddylation and inhibition of cullin-RING ubiquitin ligase (CRL) activity. CRLs are enzyme complexes that control the ubiquitination and degradation of proteins with important roles in cell cycle and survival. Potent antitumor activity in animal models correlates with accumulation of substrates of the CRLs associated with DNA replication (e.g.Cdt-1), stress response (e.g.Nrf-2), and signal transduction (e.g. pIκB). Methods: A primary objective of this phase I trial in patients (pts) with advanced non-hematologic malignancies was to evaluate the pharmacodynamic (PD) effects of MLN4924 as measured by accumulation of pIκB in peripheral blood mononuclear cells (PBMCs) and Cdt-1 and Nrf-2 in skin. Pts were treated with a daily infusion of MLN4924 for 5 days of a 21-day cycle. PBMCs were isolated for analysis of pIκB levels at screening, baseline, and at selected time points following its administration. Skin biopsies for assay of Cdt-1 and Nrf-2 were performed at baseline and following the second dose of MLN4924. Results: 10 pts have been treated at 3 dose levels. Treatment emergent adverse events include elevations in transaminases, bilirubin and alkaline phosphatase. PK analysis indicated approximately dose-proportional increases in MLN4924 exposure with an estimated half life of 5 to 15 hours and no readily apparent drug accumulation. Analysis of pIκB levels in PBMCs demonstrated accumulation from baseline levels following administration of MLN4924. Cdt-1 and Nrf-2 levels in skin rose above baseline following the second dose of MLN4924. Conclusions: Treatment of pts with MLN4924 results in accumulation of substrates associated with inhibition of NAE in both peripheral blood and skin. Inhibition occurs at exposure levels that are comparable to those resulting in pharmacodynamic activity and in vivo tumor activity in xenograft models. Evidence of PD modulation of NAE activity through the accumulation of CRL substrates has been established in this early analysis, and dose escalation is ongoing to establish the MTD of this agent.

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Phase I pharmacokinetic (PK) and pharmacodynamic (PD) study of MLN8237, a novel selective aurora A kinase (AAK) inhibitor, in patients (pts) with advanced solid tumors
Authors: Cervantes-Ruiperez A. , Elez M. E. , Roselló S. , Macarulla T. , Rodríguez-Braun E. , Lee Y. , Ecsedy J. , Liu H. , Fingert H. , Tabernero J.
Published: August 2020

2565

Background: MLN8237 is a second-generation, selective AAK inhibitor, designed to avoid benzodiazepine-like effects (somnolence) observed with another agent, MLN8054. This phase I trial examines the safety, PK, and PD of MLN8237 administered orally (PO) in pts with advanced solid tumors. Methods: Pts received MLN8237 PO once daily (QD) or twice daily (BID) for 7 days (d) followed by a 14d recovery period. Tumor and skin biopsies were done serially to evaluate bioactivity measured by mitotic index and chromosome and spindle abnormalities in mitotic cells. Doses were increased in cohorts of 3–6 pts until dose-limiting toxicity (DLT) evaluated in the first cycle was seen in ≥2 pts in a cohort. Results: As of 12-Dec-08, 27 pts received MLN8237 (median 2 cycles, range 1–10+); median age was 57 yr (range 31–78). The first cohort received 5mg QD. Higher QD doses were safe in another Phase 1 study (Infante et al, EJC Suppl 2008;6[12]:90), so subsequent cohorts in this study started at 80mg QD (Table). DLTs at 60–100mg PO BID included neutropenia, pancytopenia, stomatitis, and somnolence. Somnolence (of any grade) was seen with QD dosing, but was reduced with divided doses each <100 mg. Antitumor activity was seen in a pt in cohort 4 with a large treatment-resistant pleomorphic liposarcoma. Preliminary PK analyses showed dose-dependent increases in AUC0–24 and Cmax. Consistent with Aurora A inhibition, increased mitotic index in skin and tumor biopsies after MLN8237 dosing was coupled with decreased bipolar spindles and aligned chromosomes in mitotic cells from tumor biopsies. Conclusions: At the doses and schedule evaluated, MLN8237 is tolerable with BID dosing x7d and exhibits favorable PK, PD, and clinical antitumor activity. Additional planned cohorts include extended dosing for 14–21d in 28–35d cycles. The results support future phase 2 development.

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A phase I study of AT9283, an aurora kinase inhibitor, in patients with refractory solid tumors
Authors: Kristeleit R. , Calvert H. , Arkenau H. , Olmos D. , Adam J. , Plummer E. R. , Lock V. , Squires M. , Fazal L. , Judson I.
Published: August 2020

2566

Background: AT9283, a multitargeted kinase inhibitor, inhibits several closely related tyrosine and serine/threonine kinases with an IC50 of <10 nM including Aurora A and B, JAK and ABL. Exposure of solid tumour cell lines to AT9283 in vitro induces an “aurora inhibitory” phenotype. Cell survival decreases with increased duration of exposure. Methods: A phase I dose escalation study was performed using a 72 hour continuous intravenous (iv) infusion schedule repeated three weekly according to a stand ard “3+3” design. Results: Thirty-three patients have been treated with a median age of 61 (range 33 to 76 years). The maximum tolerated dose (MTD) was 9 mg/m2/day. Treatment was well tolerated with febrile neutropenia the only dose limiting toxicity. Other adverse events considered possibly related to AT9283 were reversible and included gastrointestinal disturbance and fatigue. Biological evidence of aurora B inhibition manifest as a reduction in histone H3 phosphorylation in skin biopsies during the infusion was observed at all dose levels. A plateau steady state plasma concentration of AT9283 was achieved within 24 hours of initiating drug infusion at all dose levels and exposure increased linearly with dose. Seven patients received an initial oral dose of AT9283 as an aqueous solution in a fasting state at a dose of 0.9 mg mg/m2 one week prior to starting iv treatment. Interim pharmacokinetic analysis indicated that the median oral bioavailability was 27% (range 17 to 45%) The best response to treatment was a partial response in one patient with NSCLC (ongoing). An additional 4 patients received at least six cycles of therapy (squamous cell carcinoma of the lung, adenocarcinoma of the esophagus and colorectal carcinoma [2]) with a best response of stable disease. Conclusions: The MTD of AT9283 when administered as a 72 hour continuous iv infusion was 9mg/m2/day. Febrile neutropenia is the dose limiting toxicity and evidence of anticancer activity was seen in heavily pre-treated patients.

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Phase I study of the novel anticancer drug PM00104 as a 24-hour IV infusion every 3 weeks (q3w) in patients (pts) with advanced solid tumors or lymphoma
Authors: Capdevila J. , Clive S. , Tabernero J. , Lardelli P. , Soto-Matos A. , Baselga J. , Piera A. , Pardos I. , Rye R. , Smyth J. F.
Published: August 2020

2568

Background: PM00104 is a novel synthetic alkaloid related to the marine compounds jorumycin and renieramycins. Preliminary preclinical studies suggest changes in cell cycle and DNA binding properties and transcriptional inhibition as main mechanisms of action. PM00104 has shown broad in vitro and in vivo anti-tumor activity (IC50 ≤ 10-8 M) with an acceptable toxicology profile. Methods: The aim of this phase I study was to assess the safety profile, dose-limiting toxicities (DLT), maximum tolerated dose (MTD), recommended dose (RD), pharmacokinetics (PK), relationship between PK and pharmacodynamics (PD) and anti-tumor activity of PM00104 administered as a 24-hour i.v. infusion q3w. Sequential cohorts of 3–6 pts were treated at 133, 266, 400, 800, 900, 1600, 3200, 4000 and 5000 μg/m2. Results: Twenty nine pts have been treated (18 male, 11 female; median age: 59, range: 44–78; ECOG PS ≤2). Five pts developed DLTs: 2 pts at 5000 μg/m2 (grade 4 thrombocytopenia/neutropenia and grade 3 nausea/vomiting in 1 pt; and grade 3 nausea in 1 pt); 1 at 4000 μg/m2 (grade 4 neutropenia/thrombocytopenia and grade 3 asthenia); 1 at 3200 μg/m2 (grade 3 tumor pain) and 1 at 266 μg/m2 (grade 3 transaminase increase). The MTD was reached at 5000 μg/m2 and the RD at 4000 μg/m2. At the RD 6 more pts have been included in order to further evaluate the safety profile and anti-tumor activity. Other adverse events included nausea and vomiting (more frequent at doses ≥800 μg/m2), fatigue, anorexia and diarrhea; most of them being of ≤grade 2 severity. No objective responses were seen but 3 pts with pancreatic adenocarcinoma, hepatocarcinoma and lower esophagus adenocarcinoma presented stable disease lasting >3 months. PM00104 shows a dose-proportional PK profile, the half-life being 20–30 hours and the volume of distribution around 1000 L. Conclusions: PM00104 has shown an acceptable safety profile with signs of anti-tumor activity in pts with advanced malignancies when administered as a 24-hour i.v. infusion q3w. PM00104 is also being evaluated with other administration schedules as monotherapy and in combination with other anti-tumor agents.

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A phase I study of flavopiridol using an alternative schedule in patients (pts) with advanced solid tumors
Authors: Ramaswamy B. , Phelps M. , Baiocchi R. , Bekaii-Saab T. , Wilkins D. , Arbogast D. , Campbell A. , Doyle A. L. , Grever M. , Shah M.
Published: August 2020

2580

Background: A phase I study of flavopiridol, a cyclin-dependent kinase inhibitor, using an alternative schedule was conducted in pts with solid tumors given its promising activity in pts with chronic lymphocytic leukemia (CLL). Methods: Using standard 3x3 ph I dose escalation design, NCI-sponsored trial was performed to determine the safety and dose-limiting toxicity (DLT) of flavopiridol given as a 30-min IV loading dose followed by a 4-hr infusion weekly for 4 wks repeated every 6 wks. DLT was defined as Gr 4 hematologic toxicity (HT) for > 7 days, > Gr 3 non-HT except Gr 3 fatigue or diarrhea resolving <4 days and cytokine release syndrome (CRS) > Gr 3 despite steroids. Blood samples were obtained at pre-dose and 0.5, 1, 3, 4.5, 6, 8, 24, and 48-hr after start of first bolus dose for pharmacokinetics (PK). Results: 26 pts with advanced solid tumors with a median age of 63 (44–75) yrs were enrolled. Median no. of doses was 7.5 (1–24). Table 1 outlines the PK parameters, DLTs and CRS. Due to a grade 5 CRS/death in cohort 3, the protocol was amended to include 20 mg IV dexamethasone prior to flavopiridol to prevent CRS (cohorts 2B, 1B). Of the 20 evaluable pts, 35% had stable- and 65% had progressive-disease. Results of serum cytokines (IL-2, IL-4, IL-6, TNF-a, IFN-g, IL-10) levels will be presented. Conclusions: There was a higher frequency of CRS, despite prophylactic steroids seen our pts with solid tumors compared to previous studies with CLL and this correlated with AUC. PK and toxicity profile in our pt population differs from pts with hematologic malignancies administered flavopiridol on the same schedule. Protein binding and serum albumin levels are under evaluation as potential contributors. This work is supported by NCI U01-CA76576.

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A phase I study of nelfinavir, an FDA approved HIV protease inhibitor, in adults with refractory solid tumors
Authors: Dennis P. A. , Blumenthal G. , Ballas M. , Gardner E. , Kawabata S. , LoPiccolo J. , Helsabeck C. , Root H. , Figg W. D. , Bernstein W.
Published: August 2020

2583

Background: Preclinical studies show that HIV protease inhibitors such as nelfinavir (N) have dose- dependent, pleiotropic anti-cancer activities. However, the maximum tolerated dose (MTD) of N has not been established in humans. Methods: Pts were treated on a modified Fibonacci dose-escalation scheme with a twice daily oral dose of N starting at the FDA approved dose of 1250 mg bid on a 21-day cycle. Patients with refractory solid tumors, ECOG PS < 2, and adequate organ function were eligible. Therapy continued until MTD or disease progression. PBMCs as well as optional tumor biopsies were collected for Akt inhibition and expression of markers of ER stress (ERS). Results: 14 patients have been enrolled. Of the 11 evaluable for toxicity, there were 10 men, 10 Caucasians, and 1 African American (median age 63 years (range 24 - 77)). Tumor types included NSCLC (3), SCLC (2), thyroid (3), pancreatic (1), colorectal (1), and renal cell (1). Median number of prior systemic therapies was 2 (range, 1 - 6). There have been no grade 4 or 5 toxicities. We observed 12 grade 3 toxicities that were asymptomatic laboratory abnormalities. The most prevalent toxicities were ALT transaminitis (5 events in 8 subjects in dose level ((DL) 2, 3, 4), AST transaminitis (4 events in 8 subjects in DL 2, 4), diarrhea (8 events in 5 subjects in DL 2, 3, 4), and hyperglycemia (5 events in 5 subjects in DL 1, 2, 3). Enrollment continues at DL 4 (3125 mg twice daily). There have been no responses. Two subjects with lung cancer had stable disease for 9 weeks. Pharmacokinetic data revealed median Cmax of 2,461.5 ng/ml (DL1), 11,809 ng/ml (DL2), 11,576 ng/ml (DL3), and 11,986 ng/ml (DL4), occurring 4 hours after an oral dose. There was no relationship between the drug levels and albumin levels. Akt inhibition and increased expression of markers of ERS and apoptosis have been observed in PBMCs from week 1 in the majority of patients at every dose level, but there was no correlation with clinical response. Conclusions: N appears to be well tolerated in subjects with advanced solid tumors at 2.5 times the FDA approved dose. AUC data suggest that there may be only minimal increases in plasma drug concentrations with doses above 1875 mg twice daily.

No significant financial relationships to disclose.
Phase II study of bavituximab plus docetaxel in patients with locally advanced or metastatic breast cancer
Authors: Tabagari D. , Nemsadze G. , Jincharadze M. , Janjalia M. , Shan J. S.
Published: August 2020

3005

Background: Phosphatidylserine (PS) is an anionic phospholipid normally present on the inside surface of cell membranes. PS becomes exposed on the outside surface of tumor endolethial cells. Bavituximab (B) is a novel anti-PS monoclonal antibody and has demonstrated preclinical synergistic antitumor activity when used in combination with docetaxel. Methods: This Simon 2-stage trial is designed to determine the overall response rate (CR+PR) in patients with locally advanced or metastatic breast cancer to a combination of weekly bavituximab (3 mg/kg) plus docetaxel (35 mg/m2) on days 1, 8, 15 of a 28-day cycle for up to 6 cycles. Following completion of docetaxel, weekly bavituximab is continued until progression or toxicity. If ≥ 6 responses are observed in the 15 pt enrolled to Stage A, an additional 31 pts will be enrolled to Stage B. Secondary objectives include time to tumor progression, duration of response, overall survival, and safety. Results: Data are available for the 15 female pts enrolled in Stage A of the trial. Median age was 49 years;13 had stage IV and 2 had recurrent/progressive stage III breast cancer at study entry. All patients received prior chemotherapy, 13 had surgery, 7 had hormone therapy and 5 had prior radiotherapy. Patients received a median of 6 cycles of treatment. All patients reported at least one AE, to date. The most common AEs (regardless of severity or causality) were alopecia, epistaxis, fatigue, back pain, diarrhea, nasal dryness, infusion related reaction, arthralgia, pyrexia, stomatitis and nail bed bleeding. Grade 3 drug-related AEs were back pain, headache, DVT (discontinued study treatment at week 3) and chest pain (SAE) starting during a docetaxel infusion, which recurred after completion of bavituximab, reported in one patient each. Using RECIST, the overall response rate in these 15 pt is 67% (95% confidence interval, 38–88%) for the intent-to-treat group and 71% (95% CI, 42–92%) for the 14 evaluable patients. All responses were confirmed at least 4 weeks later. Conclusions: Bavituximab plus docetaxel achieved an encouraging response rate in advanced/metastatic breast cancer in Stage A of this study. Enrollment in Stage B of the trial is expected to be completed with preliminary results available by the annual meeting.

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Results from the first phase I clinical study of the novel Ii-Key/HER2/neu(776–790) hybrid peptide vaccine in patients with prostate cancer
Authors: Perez S. A. , Bisias S. , Kallinteris N. L. , Ardavanis A. , Georgakopoulou K. G. , Apostolikas N. , Thanos A. , Papamichail M. , von Hofe E. , Baxevanis C. N.
Published: August 2020

3011

Background: HER-2/neu(776–790) represents an immunogenic epitope from the HER-2/neu oncoprotein whose immunogenicity is highly potentiated upon linkage with the Ii-Key moiety (LRMK) from the MHC class II invariant chain. Herein, we present the results of the first phase I clinical trial of the Ii-Key/HER-2/neu(776–790) (AE37) vaccine in patients (pts) with prostate cancer. Methods: Androgen-dependent (AD) and androgen-independent (AI) pts with HER-2/neu+ (IHC: 1–3) prostate adenocarcinomas were eligible. Concomitant medication with bicatulamide and LHRH or docitaxel was allowed. All pts received 6 monthly vaccinations with the AE37 vaccine (500 μg of AE37 plus 125 μg of GM-CSF) administered in two doses intradermally 5cm apart in the same extremity for each vaccination cycle. Immunologic responses were measured monthly in vitro by the IFNγ-based ELISPOT assay using pts’ PBMC and in vivo at the beginning and end of immunizations using DTH. Local dermal reactions were also measured after each vaccination. Results: Eligible pts [AD (n=18), AI (n=10)] were at stages T1–3N0M0, GS: 3–7 (n=18); T1–3N+M0, GS: 6–7 (n=2); T1–3N0M+, GS: 6–9 (n=4) and T1–3N+M+, GS: 6–9 (n=4). All pts had standard treatment prior to vaccinations, including surgery (S) (n=9); hormonal treatment (HT) (n=4); S+HT (n=6); S+HT+radiotherapy (RT); (n=2); S+chemotherapy (CH) (n=2); HT+RT (n=2); CH (n=3) and S+HT+CH (n=2). During vaccinations, 11 pts were free of any treatment, while 5 pts who had progressive disease received additional chemotherapy; the remainder received HT alone or combined with RT. 25 pts completed all vaccinations. Toxicity and side effects beyond grade-2 were not observed. GM-CSF was reduced by 50% for subsequent vaccinations when a local reaction of 100mm or greater was observed. DTH reactions to the parent HER-2/neu(776–790) peptide were increased (compared to pre-vaccination) for all pts, while approximately half the pts responded with significantly increased IFNγ responses to AE37 and/or parent HER-2/neu(776–790) peptide in at least 2 sequential vaccination cycles. Conclusions: The AE37 vaccine is safe and well tolerated. AE37 is also capable of eliciting potent and specific immunologic responses in prostate cancer pts.

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Correlation of immune responses and survival in a phase II study of belagenpumatucel-L in non-small cell lung cancer
Authors: Fakhrai H. , Tong A. , Nemunaitis J. , Shawler D. L.
Published: August 2020

3013

Background: Belagenpumatucel-L (Lucanix), a therapeutic vaccine comprised of 4 TGF-β2 antisense gene-modified allogeneic non-small cell lung cancer (NSCLC) cell lines, was tested in a phase II trial in which subjects with stage IIIB/IV NSCLC received intradermal injections of the vaccine every 4–8 weeks. Methods: We performed 2 cellular immunity assays and 2 humoral immunity assays on subjects in the trial. To measure the overall immune status, pretreatment peripheral blood mononuclear cells (PBMC) were stimulated by PMA/ionomycin and assessed by formation of IFN-γ ELISPOT. We also measured in vitro IFN-γ ELISPOT formation by subjects’ PBMC following vaccination. The two humoral assays looked at the induction of anti-belagenpumatucel-L antibodies and anti-HLA antibodies. Results: Subjects were classified as immune response positive (IR+) if they were positive for 3 or 4 of the assays described above. The remaining subjects were classified as immune response negative (IR-). There were 11 subjects who were IR+ and 24 who were IR-negative. The IR+ subjects had a median survival of 32.5 months (95% CI, 25.2 to 39.8) compared to 11.6 months (95% CI, 5.6 to 17.6) for IR- subjects (p=0.011, log-rank test). Subjects were also classified as clinical responders or nonresponders based on their tumor assessment after 4 months on trial. 10/19 subjects classified as clinical responders were IR+ compared to only 1/16 nonresponders (p=0.004, Fisher's exact test). Conclusions: These data demonstrate a strong correlation between the achievement of a combination of cellular and humoral immune responses and a significant increase in overall survival in the subjects treated with belagenpumatucel-L and suggest that both components of the immune system are important for the induction of clinically significant antitumor immunity by the vaccine.

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A phase I study of AS1409, a novel antibody-cytokine fusion protein, in patients with malignant melanoma (MM) or renal cell carcinoma (RCC)
Authors: Spicer J. F. , Jameson M. B. , Savage P. , Jodrell D. , Rudman S. M. , Erlandsson F. , Acton G. , McKeage M.
Published: August 2020

3024

Background: AS1409 (huBC1-huIL-12) is a fusion protein combining a humanized antibody specific for the ED-B splice variant of fibronectin with human IL-12. ED-B-fibronectin is expressed in tumor vasculature. IL-12 stimulates T and NK cell activity. Responses to IL12 occur in MM and RCC, but with high systemic toxicity. AS1409 is designed to target IL-12 to tumor vasculature. Methods: Patients with MM or RCC were treated in a dose-escalating trial of weekly i.v. AS1409 for 6 weeks with a starting dose of 15mcg/kg. Patients without unacceptable toxicity or disease progression could continue therapy. IFN-gamma and IP-10 were measured as biomarkers of activation of cellular immunity. Results: 13 patients (9 males; median age 53 years; 11 MM, 2 RCC) were treated (7 at 15mcg/kg, 6 at 25mcg/kg). DLTs observed at 25 mcg/kg were transaminase elevation and fatigue. Other toxicities included flu-like syndrome, fever, myalgia, and mucositis. Three patients continued to receive AS1409 beyond 6 weeks and 1 patient remained on treatment beyond 30 weeks. All patients showed elevation of IFN-gamma and IP10 following the first dose, although subsequently attenuated; prominent anti-drug antibody (ADA) responses were seen. A partial response was seen in a patient with MM metastatic to lymph nodes treated at 15mcg/kg, and a best response of stable disease was seen in 4 patients. Mean AS1409 half-life was 19.3 ±5.3h, mean distribution volume was 0.25 ± 0.098L/kg and clearance was 9.8 ± 6.5mL/hr/kg. Conclusions: At 15mcg/kg AS1409 was well tolerated. Biomarker activation and objective radiological evidence of anticancer activity was observed at this dose. Further study of AS1409 is focussed on optimizing dosing and scheduling, characterizing the ADA response, and antibody biodistribution.

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Novel monoclonal antibody that enhances natural killer (NK) cell cytotoxicity against multiple myeloma (MM): Preclinical data and interim phase I clinical trial results
Authors: Benson D. M. , Romagne F. , Squiban P. , Wagtmann N. , Farag S. , Mita A. , Hofmeister C. , Smith M. , Bakan C. , Caligiuri M.
Published: August 2020

3032

Background: MM is increasing in incidence and remains incurable. NK cells have modest killing activity against MM cells in part because of inhibitory signals from HLA class 1 antigens which act via the KIR receptors on NK cells. A novel anti-KIR blocking antibody (1–7F9 named IPH 2101) enhances patient NK cell cytotoxicity against autologous MM tumor cells in vitro and appears safe in an ongoing phase 1 clinical trial. Methods: NK cells from healthy controls or patients were pre-treated with IPH 2101 or IgG4 isotype control and co-cultured with MM cell lines or autologous MM tumor targets. NK cell production of interferon-gamma (IFN-γ) or granzyme B (GrB) were measured by ELISPOT. An open-label, single-agent, phase 1 dose escalation study of IPH 2101 is being conducted in patients with relapsed/refractory MM. KIR binding, pharmacokinetics, pharmacodynamics, effects on NK cell maturation, and biological effects of IPH 2101 are being monitored in all patients. Results: At an effector to target (E:T) ratio of 1:1, IPH 2101 significantly enhances NK cell IFN-γ release against MM targets (mean 33 spots/well ± 12, SEM vs. 11 ± 0.3, p = 0.005). At an E:T ratio of 10:1, IPH 2101 enhances NK cell cytotoxicity, by GrB release, of patient NK cells against autologous MM tumor cells (mean 111 spots/well ± 14, SEM vs 56 ± 10, p = 0.002). By Western blot, IPH 2101 may reduce levels of src, a kinase known to be involved in inhibitory KIR signaling. Dose escalation in the phase 1 study has been completed from 0.0003 mg/kg to 0.075 mg/kg in 14 evaluable patients. At the highest dose tested, KIR occupancy has been detected at a mean 95% ± 1.4 at 2 hours post dose, lasting up to 56% ± 18 during 2 weeks post dose. At this dose level, PK data show good correspondence with previous modeling activity. No deleterious effect on NK cell maturation has been seen. IPH 2101 has been well tolerated to date. Conclusions: IPH 2101 improves autologous NK cell killing of MM tumor cells by blocking inhibitory KIR. In the ongoing clinical trial, the antibody appears safe and well tolerated at the doses tested. This immunotherapeutic approach may hold promise as treatment for MM and further study is warranted.

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Tremelimumab in combination with exemestane as novel immunotherapy for patients with advanced breast cancer
Authors: Vonderheide R. H. , LoRusso P. M. , Khalil M. , Heath E. , Khaira D. , Soulieres D. , Dorazio P. , Mariani G. L. , Usari T. , Domchek S. M.
Published: August 2020

3034

Background: Tremelimumab (CP-675, 206) is a fully human IgG2 anti-CTLA4 monoclonal antibody (mAb) with single-agent antitumor activity presumed to be related to immune stimulation following blockade of CTLA4. Here, we conducted a phase I, open-label, dose-escalation trial of the combination of tremelimumab and exemestane in patients (pts) with advanced hormone receptor (or unknown receptor status if prior response with hormone therapy) breast cancer. Methods: Twenty-five pts with ER+ and/or PR+ breast cancer and 1 pt with unknown receptor status who had relapsed after treatment for advanced disease were enrolled. Pts received escalating doses of tremelimumab (3 to 10 mg/kg) delivered either every 28 days (Q28D) or every 90 days (Q90D) in combination with continuous exemestane (25 mg/day). Primary endpoints were assessment of safety and determination of recommended phase II dose. Tumor response evaluation per RECIST was a secondary endpoint. Results: Overall, 65 cycles of tremelimumab were administered in combination with exemestane. Pts received 3 mg/kg tremelimumab Q28D (n=6), 6 mg/kg tremelimumab Q28D (n=1), 6 mg/kg tremelimumab Q90D (n=13), or 10 mg/kg Q90D (n=6). No pharmacokinetic interaction between tremelimumab and exemestane was observed. Dose-limiting toxicities included transient serum transaminase elevation in cycle 1 (grade 3 in 1 pt) and diarrhea in cycle 1 (grade 3 in 3 pts, 1 of whom was hospitalized with steroid-refractory diarrhea and given anti-tumor necrosis factor-α mAb, infliximab) and cycle 2 (grade 3 in 1 pt). One pt developed grade 2 autoimmune thyroiditis requiring chronic hormone replacement. Other toxicities included constipation (n=6), fatigue (n=8), and rash (n=9). The MTD was 6 mg/kg Q90D. For pts receiving ≥2 cycles of therapy, best response was stable disease in 8 pts lasting 3 to 14 months (median 8 months); no pts had objective responses. Conclusions: MTD of tremelimumab in combination with exemestane is estimated at 6 mg/kg Q90D, which is lower than the single-agent dose of 15 mg/kg Q90D used in phase II and III trials in pts with melanoma or colorectal cancer. This may suggest that there is a biological interaction between tremelimumab and exemestane.

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Safety of catumaxomab: Cytokine-release-related symptoms as a possible predictive factor for efficacy in a pivotal phase II/III trial in malignant ascites
Authors: Bokemeyer C. , Heiss M. , Gamperl H. , Linke R. , Schulze E. , Friccius-Quecke H. , Lindhofer H. , Parsons S.
Published: August 2020

3036

Background: As a targeted immunotherapy, the trifunctional antibody catumaxomab specifically binds to epithelial cell-adhesion molecule (EpCAM) on tumor cells and attracts T-cells via CD3 and accessory cells via its FcγRI/III+ region. The clinical relevance of this mode of action was demonstrated by Parsons et al. (ASCO 2008) in a pivotal phase II/III trial. Catumaxomab treatment resulted in a clinically relevant prolongation of puncture-free survival (defined as time to next puncture or time to death, which ever occurred first) in patients with malignant ascites. One of the most common adverse events associated with catumaxomab is cytokine-release-related-symptoms (CRRSs), which might be of predictive value for efficacy. Methods: The safety profile of catumaxomab was identified in the pivotal study. CRRSs (pyrexia, nausea, and vomiting) were among the most common adverse events in the catumaxomab group. A correlation between CRRSs and puncture-free survival, the primary endpoint, was performed to investigate whether patients who had CRRSs benefited from catumaxomab treatment more than those who had no CRRSs. Results: Among 157 patients, CRRSs were limited to the duration of catumaxomab treatment, with a median onset of 1 day after catumaxomab administration and a median duration of 1–2 days. Most of these symptoms were mild to moderate and were manageable by standard symptomatic treatment. Only 6 of 101 patients with CRRSs who were treated with concomitant medication required systemic corticoid therapy. Puncture-free survival was longer (48 days) in patients with (81%) CRRSs during catumaxomab treatment than in patients without (19%) CRRSs (27 days), although the difference was not statistically significant (p=0.1546, log rank test). Conclusions: CRRSs are a common occurrence with catumaxomab, due to its mode of action, during the treatment of patients with malignant ascites. Since CRRSs appear to be correlated with efficacy, they may therefore be a predictive factor for catumaxomab efficacy.

[Table: see text]
Targeting of glioblastoma with activated T cells armed with anti-CD3xanti-HER2/neu (HER2Bi) and anti-CD3xanti-EGFR (EGFRBi) bispecific antibodies
Authors: Zitron I. M. , Norkina O. , Al-Kadhimi Z. , Barger G. R. , Lum L. G. , Mittal S.
Published: August 2020

3038

Background: Malignant gliomas are the most common primary brain tumors in adults. The prognosis for patients with glioblastoma remains poor despite aggressive multimodality treatment including surgery and chemoradiotherapy. The receptor tyrosine kinases EGFR, mutant EGFR (EGFRvIII), and HER2/neu are expressed on the majority of glioblastomas and are potential targets for activated T cells (ATCs) armed with bispecific antibodies (BiAbs). Methods: ATCs were generated from human peripheral blood mononuclear cells (PBMC) by culture for 14 days with monoclonal anti-CD3 and interleukin-2 and armed with HER2Bi and/or EGFRBi. HER2Bi- and/or EGFRBi-armed ATCs were examined for in vitro cytotoxicity (MTT and 51Cr release assays) against long-term malignant glioma lines (U87MG, U118MG, and U251MG) as well as primary glioblastoma lines derived from surgical specimens. Expression of EGFR and HER2/neu were evaluated by FACS. Anti-CD133 coated magnetic microbeads were used to separate CD133-positive and CD133-negative cell populations. Results: EGFRBi-armed ATCs killed up to 85% of U87, U118, and U251 targets. HER2Bi-armed ATCs exhibited comparable cytotoxicity against U118 and U251, but did not kill HER2-negative glioma U87. Cytotoxicity exhibited by either HER2Bi- or EGFRBi-armed ATCs against four primary glioblastoma cell lines was 50–80%. We found that both CD133-negative and CD133-positive cell populations were susceptible to killing by armed ATCs. When we armed ATCs simultaneously with HER2Bi and EGFRBi, killing by doubly armed ATCs was equal to or greater than that by EGFRBi-armed ATCs against the tested cell lines. Conclusions: BiAbs efficiently target ATCs to kill EGFR and/or HER2/neu expressing glioblastomas. Long-term malignant glioma cell lines and primary lines derived from surgical specimens are equally susceptible. Both CD133-negative and CD133-positive (the putative glioma stem cells) are killed. ATCs armed with BiAbs represent a potentially valuable adjuvant to current treatment.

No significant financial relationships to disclose.
Investigation of constitutive and cytokine-induced expression of pSTAT 1, 3, and 5 in PBL of breast cancer patients
Authors: Konjevic G. , Stankovic S. , Srdic T. , Jelic S. , Inic M.
Published: August 2020

3042

Background: Although cytokines mediate their effects by latent transcription factors, STATs, their dysregulation has not been much investigated in peripheral blood lymphocytes (PBL) of breast cancer patients. As alterations in the JAK-STAT pathway contribute to immunosuppression in cancer patients in this study, aside from baseline-level, we investigated the level and cytokine-mediated activation of STAT1, 3, and 5 in healthy controls and breast cancer patients. Methods: PBL (5x106/ml) of breast cancer patients in all four clinical stages, as well as healthy controls, were treated in vitro with culture medium (CM) alone, IFNα (250 IU/ml CM) and IL-2 (500 IU/ml) for 24h at 37°C. Induction of phosphorylated STAT proteins was assessed in cellular lysates by Western blotting using monoclonal anti-STAT1, 3, and 5 antibodies. Treated PBL were also analyzed for IFNγ using an intracellular flow cytometry assay. Results: Our results indicate that untreated PBL of breast cancer patients, compared to healthy controls, express a significantly lower baseline level of pSTAT1, 3, and 5. The induction of pSTAT1 expression by IFNα, similar to the induction of pSTAT3 and pSTAT5 by IL-2, is significant for healthy controls and breast cancer patients, although the effect in patients is significantly below that for healthy controls. Moreover, the induction of pSTAT5 by IL-2 is of a higher magnitude than pSTAT3. PBL of breast cancer patients, compared to healthy controls, have a lower level of IFNγ. The production of IFNγ in PBL is enhanced by IFNα, as well as IL-2, although in breast cancer patients to a much lower level than in healthy controls. Conclusions: These results provide evidence that evaluation of STAT expression in PBL of breast cancer patients could be of diagnostic and therapeutic significance for cytokine-based therapy.

No significant financial relationships to disclose.
Association of immune-regulatory (FoxP3+)-T-cell tumor infiltration status with benefit from chemoimmunotherapy with gemcitabine, oxaliplatin, 5-FU/FA plus GM-CSF and aldesleukine (GOLFIG) in metastatic colon cancer patients
Authors: Correale P. , Tagliaferri P. , Del Vecchio M. T. , Remondo C. , Migali C. , Tsang K. Y. , Rotundo M. S. , Fulfaro F. , Tassone P. , Francini G.
Published: August 2020

3045

Background: GOLFIG is a novel chemoimmunotherapy regimen, combining gemcitabine, oxaliplatin, 5-FU/FA with immunoadjuvant GM-CSF and aldesleukine, which resulted safe and very active in colon cancer patients. Antitumor activity and immunity feedback to the treatment resulted strictly correlated. The best outcome was observed in patients showing autoimmunity signs, rise in central-memory-T cells, and decline in peripheral and tumor infiltrating immuno-regulatory T (Treg) cells. On these bases, we investigated a possible correlation between Treg tumor infiltration at diagnosis and clinical outcome of these patients. Methods: An immunohistochemistry study was carried out to quantify the infiltration of Treg (FoxP3+) lymphocytes in tumor samples of 41 colon cancer patients who received FOLFOX-4 chemotherapy or GOLFIG chemo-immunotherapy as enrolled in the ongoing phase III GOLFIG-2 trial. Treg tumor infiltration score (range 0 to 5) was then correlated with survival (OS) and time to progression (TTP). Results: A higher Treg tumor infiltration score (score 3–5) was associated to a longer OS and TTP in the whole patient population (high vs low score; TTP=18 vs 9.4 months; p=0.002; OS=55.7 vs 28.9 months; p=0.001); however, those patients with high tumor infiltration of FoxP3+-T cells who received GOLFIG regimen showed the most favorable outcome (high vs low score; TTP=20.8 vs 11.6 months; p=0.04; OS=68.1 vs 41 months; p=0.04). A Cox regression model demonstrated in these patients that a high Treg tumor infiltration score is an independent variable of long survival and prolonged TTP. Conclusions: Our results suggest that GOLFIG chemoimmunotherapy is highly effective in colon carcinoma patients with high FoxP3+ infiltration score and that Treg-tumor infiltration score may be a favorable prognostic marker in colon cancer patients.

No significant financial relationships to disclose.
MMP-2: A novel target for antitumor immune therapy
Authors: Godefroy E. , Guilloux Y. , Jotereau F. , Bhardwaj N.
Published: August 2020

3053

Background: Over the last two decades, a large array of antigens expressed by melanoma cells (MAA) has been identified and used in various immunization strategies to treat cancer patients. However, the therapeutic efficacy of these approaches remains limited. Because MMP-2 (matrix metalloproteinase-2) activity is critical for melanoma progression, this protein represents a unique antigen for vaccine therapy. Methods: Antigen-specific T cell clones were obtained from healthy donors. CD4+ cells were stimulated with overlapping MMP-2 peptides pulsed with irradiated autologous CD4- fraction. After 12 days, MMP-2-specific responses were determined by intracellular staining for cytokines. Specific T cells from positive samples were enriched, using the IFNγ-capture assay (Miltenyi), followed by cloning using limiting dilution. Cytokine production was assessed by ELISA, intracellular staining, or cytometric bead array method. Results: We identified 11 novel CD4 MMP-2-derived epitopes, and showed that the corresponding T cells displayed an inflammatory TH2 profile, i.e., mainly secreting TNF, IL-4 and IL-13 and expressing the TH2-specific transcription factor GATA-3. In contrast, CD4+ T cells that recognize other associated MAAs were typically TH1. MMP-2-specific responses were detected in the CD45RO+/CD62L- T cell memory fraction. Interestingly, naïve T cells primed with active MMP-2 gave rise to TH2 specific cells, while naïve T cells primed with peptides generated IFNγ-secreting specific cells, consistent with a TH1 profile. Accordingly, we showed that the enzymatic activity of MMP-2 inhibited IL-12 secretion by mature DCs, possibly explaining the TH2 skewing of MMP-2-specific T cells. Furthermore, MMP-2 specific TH2 cells were found in at least 18 tumor-infiltrating lymphocyte populations out of the 31 melanoma patients tested. Conclusions: The protease activity of MMP-2 blocks DC maturation, more precisely IL-12 production, which could be responsible for the observed TH2 lineage commitment of MMP-2-specific T cells. These findings open the way to therapeutic strategies skewing these already frequent and diverse MMP-2-specific responses towards a more efficient antitumor TH1 phenotype to treat melanoma patients.

No significant financial relationships to disclose.
T-lymphocyte responses to the human telomerase reverse transcriptase (hTERT) in patients (pts) with advanced non-small cell lung cancer (NSCLC)
Authors: Fabre E. , Medioni J. , Dosset M. , Dosset M. , Tartour E. , Oudard S. , Riquet M. , Adotevi O.
Published: August 2020

3061

Background: hTERT is a potential target for cancer immunotherapy because it is highly expressed in tumor cells. To assess the applicability of hTERT-based immunotherapy in NSCLC, we aimed to analyse the natural anti-hTERT T-lymphocyte responses in advanced NSCLC pts. Methods: We included in a prospective, monocentric study chemonaive NSCLC pts. Pts were required to present stage III or IV tumor, without any immune deficiency or immunosuppressive drug. Before start of chemotherapy, the anti-hTERT T-lymphocyte responses were assessed in whole blood by ELISPOT and thymidine proliferation test. We evaluated the presence or absence of hTERT-specific T lymphocytes. Thereafter, we analyzed its link with age (< 60 vs. ≥ 60 yrs), ECOG performance status PS (0–1 vs. 2–3), tumoral stage (III vs. IV), histological subtype (adenocarcinoma vs. other), and smoking status (never smoker vs. smoker). Statistical analysis was performed using chi-square or Fischer test. Results: Between February and September 2008, 31 NSCLC pts were included. They presented a stage III (n = 6) or IV (n = 27) disease. Median age was 65 yrs (33–89). Eighteen pts presented anti-hTERT T lymphocytes (54%); among them we found 12 pts ≥ 60 yrs (70%), 14 adenocarcinoma (77%), 13 stage IV (72%), 7 pts with PS 2/3 (41%), and 13 smokers (72%). So far, univariate analysis showed no relationship between presence of anti-hTERT T-cell responses and pts’ characteristics. Conclusions: These preliminary results demonstrate that natural anti-hTERT immune response exists in NSCLC pts, even in case of poor prognosis. Thus, clinical trial using telomerase-based therapeutic vaccine may include NSCLC pts with advanced disease. The study is ongoing to evaluate if baseline anti-hTERT immune response could be used as selection criteria for telomerase vaccination in NSCLC.

No significant financial relationships to disclose.
Evaluation of recombinant bacillus Calmette-Guerin expressing pertussis toxin in bladder cancer
Authors: Chade D. C. , Borra R. , Leite K. R. , Villanova F. E. , Nascimento I. , Liete L. C. , Srougi M. , Andrade P. M.
Published: August 2020

3064

Background: Pertussis toxin (S1PT) was combined to BCG, creating a recombinant variant (rBCG-S1PT) with the capacity to promote an immune response targeted to the T helper type 1 (Th1), which may increase the effectiveness of its antitumor effect. Objective: Compare the antitumor effects of rBCG-S1PT and BCG in an experimental model of bladder cancer. Methods: The development of the animal model of bladder cancer was conducted by transurethral instillation of bladder tumor cell line MB49 of the mouse strain C57BL/6. The animal models were divided into three groups, which received 4 weekly intravesical applications of rBCG-S1PT, BCG, or saline. After 7 days of the last instillation, bladders were extracted and weighed in order to infer the tumor weight. Then, the bladders were divided for molecular analysis to assess the pattern of immune response and histopathological analysis. In another experiment, animals were monitored for 60 days for analysis of survival. Also, a test was carried out as previously described, but with no tumor cells instillation. Results: The rate of tumor implantation was 90% of the animals submitted to tumor inoculation. We obtained reduction of the average weights of bladder in groups BCG and rBCG-S1PT ((p<0.001). In both groups treated with immunotherapy, there was an increase of expression of interleukins related to Th1, which was more intense in the group treated with rBCG-S1PT (p<0.05). The analysis of survival showed a significant increase in the group of animals treated with rBCG-S1PT. The instillation of immunotherapeutic agents in animals without tumor did not demonstrate histological differences when compared to the control group. Conclusions: The establishment of the syngeneic orthotopic animal model was successful. The immunotherapy with rBCG-S1PT demonstrated more benefits than BCG in the treatment of bladder cancer in mice, reducing the bladder weight and increasing survival. The immune response obtained with the rBCG-S1PT expressed higher cytokines related to Th1. This new recombinant agent may promote better bladder tumor control than BCG imunotherapy in a clinical setting.

No significant financial relationships to disclose.