Human Anti-human Antibody Research Articles

Array-in-well serodiagnostic assay utilizing upconverting phosphor label technology

In this study, a multiplex serological array-in-well assay was constructed for simultaneous detection of serum IgG antibodies against parvovirus B19 and human adenovirus. The array was prepared in streptavidin-coated 96-well microtiter plates by spotting biotinylated parvovirus B19 virus-like-particles, adenovirus type 2 and 5 hexon antigens, negative control of human serum albumin and positive controls of human IgG and anti-human IgG antibodies on the bottom of each well in an array format with a printable area of 2mm×2mm. The array-in-well assay was evaluated with serum samples (n=89) of different antibody status as determined by commercial enzyme immunoassay for parvovirus IgG, and by in-house enzyme immunoassay for adenovirus IgG. The bound serum anti-parvovirus IgG, anti-adenovirus IgG, and total IgG antibodies were detected with anti-human IgG antibody coated photon upconverting nanoparticles and the assay was measured with an anti-Stokes photoluminescence imager. Detection of specific antibodies by the multiplex array-in-well assay was in good agreement (100% for parvovirus B19 and 96% for adenovirus) with the reference results. In conclusion, the array-in-well with upconverting phosphor reporter technology was able to detect antiviral antibodies in human sera, and represents an efficient serodiagnostic concept that is a promising new tool for multiplex serology.

The history of the research and development of human anti-human PD-1 antibody nivolumab as a new immune checkpoint inhibitor

The history of the research and development of human anti-human PD-1 antibody nivolumab as a new immune checkpoint inhibitor

Pharmacological profile and clinical efficacy of human anti-human PD-1 antibody nivolumab (OPDIVO®) as a new immune checkpoint inhibitor

ニボルマブ（遺伝子組換え）［商品名：オプジーボ®点滴静注20 mg，100 mg，以下ニボルマブ］は，ヒト型抗ヒトPD-1モノクローナル抗体であり，「根治切除不能な悪性黒色腫」を効能・効果として，2014年9月より世界に先駆けて日本で発売された新たな免疫チェックポイント阻害薬である．非臨床試験において，ニボルマブは，ヒトPD-1の細胞外領域に特異的に結合し，PD-1とPD-1リガンド（PD-L1およびPD-L2）との結合を阻害した．また，ニボルマブは抗原刺激によるヒトT細胞の増殖およびIFN-γ産生を増強し，悪性黒色腫患者T細胞を用いた悪性黒色腫抗原ペプチド再刺激系においては，腫瘍抗原特異的CD8陽性T細胞およびIFN-γ産生細胞を増加させ，ヒト悪性黒色腫細胞に対するCD8陽性T細胞の細胞傷害活性を増強した．さらに，ニボルマブは各種抗原を接種したサルの細胞性および液性免疫応答を増強し，抗マウスPD-1抗体4H2は，マウス同系担がんモデルにおいて腫瘍組織中の免疫関連遺伝子の発現量を増加させ，抗腫瘍効果を示した．このようにニボルマブは，PD-1とPD-1リガンドとの結合を阻害し，抗原特異的なT細胞の増殖，活性化およびがん細胞に対する細胞傷害活性を増強することで抗腫瘍効果を示すことから，悪性腫瘍に対する新たな治療薬になると考えた．臨床試験において，悪性黒色腫患者を対象とした国内第Ⅱ相試験にて有効性，安全性および忍容性が確認されたことから，ニボルマブは悪性黒色腫の有用な治療薬となりえることが示された．本試験結果を踏まえ，2013年12月に製造販売承認申請を行い，2014年7月にニボルマブは世界初の抗PD-1抗体として製造販売承認を取得した．現在，様々ながん腫に対するニボルマブの臨床試験が実施されており，今後ニボルマブが，がん治療の選択肢を広げるものと期待される．

458P - Phase I Study of Ontuxizumab, a Humanized Monoclonal Antibody Recognizing Endosialin/Tumor Endothelial Marker1 (Tem1), in Japanese Patients with Solid Tumor

ABSTRACT Aim: Endosialin is a cell surface glycoprotein that is expressed on cells involved in the development of tumor vasculature, primarily pericytes and stromal fibroblasts. Ontuxizumab is humanized immunoglobulin G-1-kappa (IgG1/k) monoclonal antibody that is the first clinical-stage agent to target endosialin. The antibody has shown antitumor activity in a variety of models and its pharmacologic activity have shown distinct non-disease activities as compared to other anti-vascular therapies, such as no observed impact on wound healing. Methods: Patients (pts) with solid tumor who have no other appropriate treatment were eligible in the study. Each cycle consisted of four weeks. Ontuxizumab was administered weekly until disease progression or occurrence of a DLT and administered to 4 cohorts at 2, 4, 8 and 12 mg/kg weekly schedule in a stepwise dose escalation manner. Results: Fifteen pts were enrolled including three gastric cancer, two gastrointestinal stromal tumor, two breast cancer and a patient with lung cancer, malignant pheochromocytoma, adrenocortical cancer, hypopharyngeal cancer, extraskeletal chondrosarcoma, thymic carcinoma, renal pelvis cancer and intrahepatic cholangiocarcinoma. No DLT was observed. Treatment related adverse events were observed in 5 of 15 pts and the most common events were grade 1 constipation (two pts), grade 2 hyperkalaemia (two pts), grade 2 infusion related reaction (two pts). Cmax and AUC values of ontuxizumab after single administration increased in a dose-related manner. Mean t1/2 was 85.2 - 219 hours after multiple administrations. Human anti-human antibody was not detected. Of eight pts with stable disease, one pt with GIST after failure of imatinib and sunitinib showed tumor shrinkage by CT evaluation based on Choi's criteria at 12 mg/kg. The longest duration of ontuxizumab treatment was 32 weeks for one gastric cancer pt in 4 mg/kg. Conclusions: Ontuxizumab was well tolerated in pts with advanced solid tumors at up to 12 mg/kg. Ontuxizumab is currently being investigated in Ph2 studies for its potential treatment of melanoma, metastatic colorectal carcinoma and soft tissue sarcoma. Disclosure: T. Doi: Other substantive relationships:Participant in Eisai sponsored speaker's bureau; Y. Naito: Other substantive relationships:Participant in Eisai sponsored speaker‘s bureau; M. Tahara: Advisory Board: Eisai; K. Nakai: Employee of Eisai. Own stock of Eisai, A. Ohtsu: Advisory Board: Eisai. All other authors have declared no conflicts of interest.

Development of a complete human anti-human transferrin receptor C antibody as a novel marker of oral dysplasia and oral cancer.

Oral squamous cell carcinoma (OSCC) is the sixth most common cancer worldwide. Up to 20% of oral dysplasia cases have been suggested to undergo malignant transformation to OSCC; however, there are no methods to predict OSCC development. In this study, to identify the genes associated with oral dysplasia progression, we performed genomic copy number analyses of genomic DNA samples isolated from primary oral dysplasia and OSCC via the microdissection method and found elevated expression of transferrin receptor C (TfR1/TFRC) with genomic amplification in oral dysplasia and OSCC. The expression rate of TFRC in OSCC was significantly higher than that in dysplasia, suggesting that OSCC disease progression might be related to TFRC expression. Additionally, we investigated the in vitro and in vivo impacts of a newly established anti-human TFRC monoclonal antibody, which was isolated from a human cDNA library using the phage-display method, on cell proliferation and survival. The anti-TFRC antibody blocked the interaction between transferrin and TFRC and consequently inhibited iron uptake, leading to the iron deprivation-mediated suppression of cell growth and induction of apoptosis. Moreover, we demonstrated that the anti-TFRC antibody efficiently inhibited tumor growth in a murine xenograft OSCC model. Therefore, we suggest our developed complete human anti-human TFRC antibody as a useful, novel treatment for oral dysplasia and OSCC.

FRI0383 Safety, Pharmacokinetics, and Pharmacodynamics of Epratuzumab in Japanese Patients with Moderate-To-Severe Systemic Lupus Erythematosus: Results from A Phase 1/2 Study

BackgroundTreatment with epratuzumab, a humanized monoclonal antibody targeting CD22 on B cells, has been associated with improvements in disease activity in patients (pts) with moderate-to-severe systemic lupus erythematosus (SLE) with...

Safety with Ocrelizumab in Rheumatoid Arthritis: Results from the Ocrelizumab Phase III Program

ObjectiveThe objective was to determine the safety of ocrelizumab (OCR) in patients with rheumatoid arthritis (RA).MethodsThis was an analysis of the double-blind, placebo-controlled periods and long-term follow-up of 4 OCR phase III trials in RA (SCRIPT, STAGE, FILM and FEATURE). Safety data per study and the results of a meta-analysis of serious infectious events (SIEs) are presented.ResultsOverall, 868 patients received placebo, 1064 patients OCR 200 mg×2 (or 400 mg×1) (OCR200), and 827 patients OCR 500 mg×2 (OCR500) plus background methotrexate (MTX) at baseline and 24 weeks. During the double-blind, placebo-controlled periods, the incidence of adverse events and serious adverse events was comparable between the OCR+MTX and placebo +MTX groups. Infusion-related reactions were more common with OCR+MTX and decreased in frequency with subsequent infusions. Serious infusion-related reactions were rare (0.1%). Serious infections occurred more frequently with OCR500+MTX. In the meta-analysis, a statistically significant difference from placebo +MTX in incidence of SIEs per 100 patient-years of 2.4 (95% CI, 0.3–4.5) was observed with OCR500+MTX, but not with OCR200+MTX (0.6; 95% CI, −1.3 to 2.4). Patients recruited in Asia exhibited a higher risk of serious infections (hazard ratio, 1.78; 95% CI, 1.03–3.06). The incidence of human anti-human antibodies was <5%. Long-term follow-up indicated no differences in malignancy rates between the treatment groups. There was no apparent difference in time to B-cell repletion between the OCR dose groups.ConclusionsIn placebo-controlled clinical trials of RA, OCR500+MTX was associated with a higher risk of serious infections compared with placebo +MTX. The safety profile of OCR 200+MTX was comparable with placebo+MTX.Trial RegistrationSTAGE Clinical Trials.gov NCT00406419 SCRIPT Clinical Trials.gov NCT00476996 FILM Clinical Trials.gov NCT00485589 FEATURE Clinical Trials.gov NCT00673920

Safety and efficacy of ofatumumab in relapsing-remitting multiple sclerosis

We present the first study to explore safety and efficacy of the human CD20 monoclonal antibody ofatumumab in relapsing-remitting multiple sclerosis (RRMS). In this randomized, double-blind, placebo-controlled study, patients received 2 ofatumumab infusions (100 mg, 300 mg, or 700 mg) or placebo 2 weeks apart. At week 24, patients received alternate treatment. Safety and efficacy were assessed. Thirty-eight patients were randomized (ofatumumab/placebo, n = 26; placebo/ofatumumab, n = 12) and analyzed; 36 completed the study. Two patients in the 300-mg group withdrew from the study because of adverse events. No unexpected safety signals emerged. Infusion-related reactions were common on the first infusion day but not observed on the second infusion day. None of the patients developed human anti-human antibodies. Ofatumumab was associated with profound selective reduction of B cells as measured by CD19(+) expression. New brain MRI lesion activity was suppressed (>99%) in the first 24 weeks after ofatumumab administration (all doses), with statistically significant reductions (p < 0.001) favoring ofatumumab found in new T1 gadolinium-enhancing lesions, total enhancing T1 lesions, and new and/or enlarging T2 lesions. Ofatumumab (up to 700 mg) given 2 weeks apart was not associated with any unexpected safety concerns and was well tolerated in patients with RRMS. MRI data suggest a clinically meaningful effect of ofatumumab for all doses studied. Results warrant further exploration of ofatumumab in RRMS. This study provides Class II evidence that in patients with RRMS, ofatumumab compared with placebo does not increase the number of serious adverse events and decreases the number of new MRI lesions.

Abstract P4-16-09: Phase 1b/2 trial of the HER3 inhibitor patritumab (U3-1287) in combination with trastuzumab plus paclitaxel in newly-diagnosed patients with HER2+ metastatic breast cancer (MBC)

Abstract Background: Patritumab is a fully human anti-HER3 monoclonal antibody that has shown potent antitumor activity in vivo. HER3 is a key dimerization partner for other HER family members, and studies suggest that the HER3:HER2 heterodimer is the most potent signaling pair. Therefore, combined inhibition of HER3 and HER2 may synergistically inhibit breast cancer tumor growth. In the CLEOPATRA trial, pertuzumab combined with trastuzumab and docetaxel prolonged progression-free survival compared to trastuzumab and docetaxel alone, demonstrating the benefit of comprehensive blockade of HER2 dimer signaling. Trastuzumab is approved in combination with paclitaxel for first-line treatment of HER2+ MBC. This phase 1b/2 study is investigating patritumab in combination with trastuzumab and paclitaxel in patients (pts) with newly-diagnosed MBC. Results of the phase 1b portion are reported here. Methods: Eligible pts had HER2+ newly-diagnosed MBC. In the open-label, phase 1b portion of this trial, pts received intravenous (IV) patritumab 18 mg/kg in combination with trastuzumab (8 mg/kg IV loading dose; 6 mg/kg IV maintenance dose) and paclitaxel (175 mg/m2 IV) every 3 weeks (Q3W). In the event that 18 mg/kg was not tolerated based on dose-limiting toxicity (DLT) assessment, sequential cohorts were to receive de-escalating doses of patritumab. Phase 1b study end points included adverse event (AE) incidence, human antihuman antibody (HAHA) formation, pharmacokinetics (PK), and tumor response. Results: Six pts were enrolled in the phase 1b portion of the trial, with a median age of 61 years (range, 51-78). There were no reported DLTs. Grade ≥3 treatment-related AEs occurred in 3 pts: 1 pt had a serious AE of grade 3 pneumonia; 1 pt had grade 3 worsening of arm pain; 1 pt had grade 3 oral mucositis, prolonged QTc, flu-like syndrome, and increased transaminases. In this limited pt population, most ECG changes were within or slightly above normal limits of the QTc interval. Only 2 pts had increased values close to 50 msec compared with baseline. It should be noted that baseline QTc values for 3 pts (including the pt that experienced grade 3 prolonged QTc) were close to the upper limit of normal. There were no grade 4 AEs, and no other serious AEs. All other treatment-related AEs were grades 1 or 2. All 6 pts tested negative for HAHA formation after drug administration. PK data are consistent with previous studies with patritumab. Two pts had complete response (CR) as their best overall response, 2 pts had partial response (PR), 1 pt had stable disease, and 1 pt was not evaluated for tumor response. All 6 pts have discontinued treatment; 2 due to progressive disease, 3 due to pt decision (2 with CR and 1 with PR), and 1 at the investigator's discretion. Conclusions: Results to date indicate that the combination of patritumab with trastuzumab and paclitaxel is generally well tolerated, with a promising response rate. As no DLTs were reported, the recommended phase 2 dose is patritumab 18 mg/kg with trastuzumab (8 mg/kg loading; 6 mg/kg maintenance) and paclitaxel 175 mg/m2 Q3W. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-16-09.

Impact of interleukin-1β antibody (canakinumab) on glycaemic indicators in patients with type 2 diabetes mellitus: Results of secondary endpoints from a randomized, placebo-controlled trial

Aims/hypothesisThis study was conducted to determine the optimal monthly subcutaneous dose of canakinumab (a human monoclonal anti-human IL-1β antibody) needed to improve glucose control in metformin-treated patients with type 2 diabetes mellitus (T2DM). MethodsThis was a parallel-group, randomized, double-blind, multicentre, placebo-controlled study designed to assess the effect on HbA1c and the safety/tolerability of four monthly doses of canakinumab (5, 15, 50, or 150mg) as an add-on to metformin over 4 months. ResultsPatients (n=551; mean age 54.1 years; mean baseline HbA1c 7.4%) were randomized and treated in a double-blind fashion to canakinumab 5mg (n=93), 15mg (n=95), 50mg (n=92), 150mg (n=92) or placebo (n=179) monthly. There was no dose response detected between active canakinumab doses, but all doses numerically lowered HbA1c (primary endpoint) from baseline between 0.19% and 0.31% (placebo-unadjusted), with maximal effect noted in the 50mg dose of canakinumab (−0.18% difference vs placebo; multiplicity-adjusted, P=0.13902) as reported earlier (Ridker et al., 2012). No other glycaemic control parameters (FPG, fasting insulin, plasma glucose AUC0–4h, 2-h PPG, peak glucose, C-peptide AUC0–4h, peak C-peptide, insulin AUC0–4h, peak insulin, ISR0–2h, HOMA-β and HOMA-IR) showed any meaningful changes by canakinumab therapy. Canakinumab treatment was safe and well tolerated. There were no relevant differences in adverse events between the canakinumab and placebo groups. Conclusions/interpretationA 4-month course of monthly canakinumab (50mg) produced a numerical reduction of HbA1c in T2DM patients on metformin, potentially by improving beta-cell function. The safety and tolerability profile of canakinumab was consistent with prior trials. Trial registrationRegistry: http://www.ClinicalTrials.gov, Registration No.: NCT00900146

A first-in-human trial of RG7116, a glycoengineered monoclonal antibody targeting HER3, in patients with advanced/metastatic tumors of epithelial cell origin expressing HER3 protein.

2522 Background: The Human Epidermal Growth Factor Receptor 3 (HER3) is a key heterodimerization partner for other HER family members thereby acting as a downstream signal amplifier. This is a first in human study evaluating the safety of RG7116, a humanized anti-HER3 monoclonal antibody with potent HER3 signal inhibition. Due to a glycoengineered Fc-part this antibody displays enhanced antibody-dependent cellular cytotoxicity as compared to conventional antibodies. Methods: Patients (pts) with advanced or metastatic carcinomas with centrally confirmed HER3 protein expression were included. A “3+3” dose escalation design was performed starting at 100 mg flat dosing in a q2w regimen. In addition to single agent RG7116 (Part A), RG7116 plus cetuximab (Part B) and RG7116 plus erlotinib (Part C) combinations are evaluated. The results of Part A are presented. Results: Twenty-five pts have been enrolled in 6 cohorts (100 to 2000 mg). No DLTs were observed. Pts had a median (range) of 3 (2 to 6) prior chemotherapy lines. Nine infusion-related reactions (IRRs) Gr 1 to 3 occurred in 7 pts. Three drug-related AEs Gr 3 were reported, 1 IRR, 1 GGT increase, and 1 neutropenia. Only 1 patient was tested positive for human anti human antibodies (HAHA). The PK of RG7116 was non-linear from 100 mg up to 400 mg, possibly as a result of target-mediated drug disposition. Both Cmax and AUC showed a greater than doseEproportional increase over the same dose range, accompanied by a decline in total clearance. Dose proportionality was observed from 800 mg onwards. PD effects were observed from the first dose level onwards with HER3 membranous protein down-regulation in skin and from 200 mg onwards in ontreatment tumor samples. Five pts (1 NSCLC, 2 CRC, 1 SCCHN, 1 BC) had a best response of SD. Confirmed SD (&gt;16 weeks) was observed in 2 pts. One BC patient achieved a PR in 18 FDG-PET and significant shrinkage of non-target tumor lesions on CT scan. Conclusions: RG7116 is the first glycoengineered monoclonal anti-HER3 antibody in clinical development. RG7116 monotherapy was well tolerated, and demonstrated preliminary signs of clinical activity. Clinical trial information: NCT01482377.

Phase I study of tremelimumab (CP-675 206) plus PF-3512676 (CPG 7909) in patients with melanoma or advanced solid tumours

Background:Tremelimumab, a fully human cytotoxic T-lymphocyte antigen 4 monoclonal antibody, and PF-3512676, a Toll-like receptor-9 agonist, are targeted immune modulators that elicit durable single-agent antitumour activity in advanced cancer.Methods:To determine the maximum tolerated dose (MTD) of these agents combined during this phase I study, patients received intravenous tremelimumab (6.0, 10.0, or 15.0 mg kg−1) every 12 weeks plus subcutaneous PF-3512676 (0.05, 0.10, or 0.15 mg kg−1) weekly. Primary end points were safety and tolerability; secondary end points included pharmacokinetics and antitumour activity.Results:Twenty-one patients with stage IV melanoma (n=17) or advanced solid tumours (n=4) were enrolled. Injection-site reactions (n=21; 100%), influenza-like illness (n=18; 86%), and diarrhoea (n=13; 62%) were the most common treatment-related adverse events (TAEs). Grade ⩾3 TAEs were reported (n=7; 33%). Dose-limiting toxicities (prespecified 6-week observation) occurred in one of the six patients in the 10 mg kg−1 tremelimumab plus 0.05 mg kg−1 PF-3512676 cohort (grade 3 hypothalamopituitary disorder) and two of the six patients in the 15 mg kg−1 tremelimumab plus 0.05 mg kg−1 PF-3512676 cohort (grade 3 diarrhoea). Consequently, 15 mg kg−1 tremelimumab plus 0.05 mg kg−1 PF-3512676 exceeded the MTD. Two melanoma patients achieved durable (⩾170 days) partial response. No human antihuman antibody responses to tremelimumab were observed.Conclusion:Weekly PF-3512676 (⩽0.15 mg kg−1) plus tremelimumab (⩽10 mg kg−1 every 12 weeks) was tolerable.

Abstract LB-159: A Phase I study of IMMU-130 (labetuzumab-SN38) anti-CEACAM5 antibody-drug conjugate (ADC) in patients with metastatic colorectal cancer (mCRC).

Abstract Background: IMMU-130 is an ADC comprising the humanized monoclonal IgG antibody (mAb), hMN-14 (labetuzumab), bound to the active metabolite of irinotecan, SN-38 (6 SN-38/IgG). hMN-14 a slowly-internalizing mAb, recognizes the CEA (CEACAM5; CD66e) antigen, which is expressed in many solid cancers, including &amp;gt;80% of CRC. hMN-14 was safe in prior clinical trials, when administered unconjugated or bound to 131-I for radioimmunotherapy. In vitro studies showed that the antibody-drug linkage was susceptible to cleavage in serum, with 50% of SN-38 released in ~1.0 day. This means that after the antibody binds to its tumor target, SN-38 is released, leading to a locally enhanced concentration within the tumor site. Irinotecan is a standard therapy of mCRC, but has major gastrointestinal and hematologic toxicity. By targeting SN-38 directly to CEA-expressing tumors, delivery of SN-38 may be increased while avoiding systemic toxicity. In animal CRC xenograft models, IMMU-130 exhibited high anti-tumor activity. A safe starting dose for humans was determined from preclinical toxicology studies that showed no gastrointestinal toxicity, and only transient mild myelosuppression. This single-arm, open-label, dose-escalation, phase I study was undertaken to define the maximum-tolerated dose (MTD) of IMMU-130 in patients with mCRC. Methods: Eligible patients were previously treated with at least one prior irinotecan-containing regimen, had serum CEA &amp;gt;5 ng/mL, and measurable disease by CT/MRI. IMMU-130 was administered every-other-week, with a goal of achieving 12 cycles (24 weeks) in the absence of unacceptable toxicity or disease progression. Pts were entered in cohorts treated at increasing dose levels, with DLT defined as Grade 4 neutropenia ≥ 5 days, ≥ Grade 3 thrombocytopenia, Grade 3 anemia, ≥Grade 3 nausea, vomiting or diarrhea persisting &amp;gt;48 hr, and other ≥ Grade 3 non-hematologic toxicity during the first two cycles. Results: So far 9 pts have been treated at the 2, 4, 8, and 16 mg/kg dose levels. Pts had received a median of 2 prior regimens. The median number of cycles given is 3.9, with 6/9 patients receiving 3 or more doses. Of 5 pts having &amp;gt;2 doses of 16 mg/kg, one has currently received 14 doses and has a continuing partial response after 8. One DLT was observed at 16 mg/kg: Grade 3 thrombocytopenia. The same pt experienced Grade 4 neutropenia &amp;lt;5 days. A second pt at the same dose also had Grade 4 neutropenia &amp;lt;5 days. No grade 2 or higher drug-related GI toxicities were observed. A single case of hypersensitivity occurred. No human anti-human antibodies (HAHA) have been detected to date. Analysis of serum samples shows the intact conjugate clears more quickly than the Ig, consistent with SN-38 being gradually released from the ADC. Conclusion: IMMU-130 appears to be safe and reasonably well tolerated within a clinically effective dosage range. Dose escalations are continuing. Citation Format: Neil H. Segal, Jaclyn Verghis, Serengulam Govindan, Pius Maliakal, Robert M. Sharkey, William A. Wegener, David M. Goldenberg, Leonard B. Saltz. A Phase I study of IMMU-130 (labetuzumab-SN38) anti-CEACAM5 antibody-drug conjugate (ADC) in patients with metastatic colorectal cancer (mCRC). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-159. doi:10.1158/1538-7445.AM2013-LB-159

Safety, pharmacokinetics and pharmacodynamics of the anti-A33 fully-human monoclonal antibody, KRN330, in patients with advanced colorectal cancer

PurposeThe objective of this first-in-human trial included the safety, maximum tolerated dose (MTD), pharmacokinetics, immunogenicity and antitumour effects of KRN330, a novel fully-human monoclonal antibody directed against A33, a membrane bound glycoprotein uniformly expressed in 95% of colorectal cancers. MethodsPatients with advanced or metastatic colorectal cancer (CRC) refractory to standard therapy were eligible. Twenty-nine patients received weekly intravenous KRN330 (0.1–10mg/kg) for a minimum of 4 weeks in a standard 3+3 design, and nine patients received q2week doses at 3mg/kg with pre- and post-biopsies to evaluate tumour binding and safety on this schedule. ResultsThe most common KRN330 related adverse events (all grades) were nausea (66%), diarrhoea (61%) and vomiting (47%). The MTD was 3mg/kg weekly, with dose-limiting grade 3 gastrointestinal toxicities at 10mg/kg and the intermediate dose level of 6mg/kg. Pharmacokinetics of KRN330 was linear. Stable disease was reported in 12/38 patients (32%), with a median duration of 155 days. There was no evidence of human anti-human antibodies, and immunohistochemistry on biopsy samples demonstrated that KRN330 remained bound to tumour 2 weeks after dosing. ConclusionsKRN330 is safe and tolerable at the MTD of 3mg/kg once weekly in patients with advanced CRC. Dosing on alternate weeks is supported by tumour binding. The long treatment durations and lack of immunogenicity warrant further investigation of KRN330 in combination.

Pharmacokinetic and Pharmacodynamic Analysis of Ocaratuzumab in 50 Patients with Relapsed Follicular Lymphoma and Low-Affinity FcγRIIIa (CD16a) Polymorphisms.

Abstract 2750Ocaratuzumab, previously known as AME-133v, is a Fc- and Fab-engineered humanized anti-CD20 monoclonal antibody designed for optimized affinity to the CD20 antigen as well as to the FcγRIIIa (CD16a) receptor on effector cells. It demonstrates 13–20 fold higher affinity for CD20 and 6-fold higher antibody-dependent cellular cytotoxicity compared to rituximab.In a Phase II study, 50 patients received IV ocaratuzumab at 375 mg/m2 weekly for 4 doses, the standard dosing schedule of rituximab. The median age (range) was 61 (39–83) years and the median number of prior systemic therapies (range) was 2 (1–9). Two patients were rituximab naïve.This abstract describes the pharmacokinetics (PK) and pharmacodynamics of ocaratuzumab. The steady state concentration of ocaratuzumab was reached by the 2nd dose and started to decline 1 week after the last dose. The PK of ocaratuzumab is best characterized by a 2-compartment model with a typical clearance estimate of 0.26 L/day. The typical volume estimates for the central and peripheral compartments were 3.15 L and 3.31 L respectively, which corresponds to a terminal half-life of approximately 19 days.As expected, there was significant inter-patient variability in the clearance of ocaratuzumab. Analysis of the serum concentration of ocaratuzumab revealed no correlation between serum ocaratuzumab concentration and progression free survival (PFS) as some patients with low concentrations had prolonged PFS and some patients with high concentrations had short PFS (Figure 1).Only 2 patients developed high titers of human anti-human antibody. These 2 patients maintained very low concentrations of ocaratuzumab and were amongst 3 total patients who demonstrated progressive disease in the study.All patients had rapid and sustained B-cell depletion for at least 90 days after the last infusion. A retrospective analysis of B-cell recovery and response showed a direct correlation between time to B-cell recovery and PFS duration. Patients with prolonged B-cell depletion stayed in remission longer than patients who recovered their circulating B-cells more quickly.In conclusion, clinical efficacy was seen across a wide range of serum concentrations of ocaratuzumab, suggesting significant potency of the drug even at low serum concentration levels. Despite all patients having low-affinity FcγRIIIa polymorphisms, the overall response rate was 32% with a median PFS of 38 weeks. Although a weekly dose of 375 mg/m2 for 4 doses was used in this trial, lower doses of ocaratuzumab, such as 100 mg/m2 given once weekly for 4 doses, may potentially be effective. [Display omitted] Disclosures:Du:Mentrik Biotech, LLC: Employment. O’Reilly:Mentrik Biotech, LLC: Employment. Le:Mentrik Biotech, LLC: Employment. Jain:Mentrik Biotech, LLC: CEO Other, Employment, Equity Ownership, Patents & Royalties.

Guidance on the use of canakinumab in patients with cryopyrin-associated periodic syndrome in Japan

Cryopyrin-associated periodic syndrome (CAPS) is an orphan disease with incidence of about one in 1,000,000 persons. This autoinflammatory disease develops in the neonatal period or early childhood, with various inflammatory symptoms occurring repeatedly throughout the patient's lifetime. It is caused by abnormality of the NLRP3 protein which mediates the intracellular signal transduction mechanism of inflammatory processes, resulting in continuous overproduction of interleukin (IL)-1β, which induces chronic inflammation and progressive tissue damage. Definitive diagnosis of CAPS is difficult, and treatment has also been difficult because of a lack of effective medications in Japan. Clinical studies of human anti-human IL-1β monoclonal antibody (canakinumab) treatment were conducted in Japan, and approval was granted for therapeutic use of canakinumab for CAPS in September 2011. Similar to other biological drugs, canakinumab is clinically highly effective. However, sufficient attention to the method of use and adverse drug reactions is necessary. This guidance describes the use of canakinumab in Japan for CAPS in relation to exclusion criteria, method of use, evaluation criteria, and adverse drug reactions.

IS6-3 - Phase I Study of Anti PD-1 Antibody Ono-4538 in Japanese Patients with Advanced Solid Tumors

ABSTRACT Background Programmed cell death-1 (PD-1) is a co-inhibitory receptor expressed on antigen-activated T cell. PD-L1, ligand of PD-1, expressed on various human tumor cells inhibits T-cell activation and proliferation by binding to PD-1. Expression of PD-L1 correlates with poor outcome in renal cell carcinoma, melanoma, non-small-cell lung cancer (NSCLC), and colorectal cancer (CRC). ONO-4538 (BMS-936558) is a fully human monoclonal IgG4 antibody binding to PD-1, and enhances antitumor activity by blocking the PD-1/PD-L1-induced down-regulation of the T-cell co-stimulatory signal. This phase I study investigated the tolerability, pharmacokinetics (PK), antitumor activity and immune response in Japanese patients with advanced solid tumors. Method Patients with advanced solid tumors refractory to standard therapy, adequate organ functions and performance status of 0 or 1 were enrolled. ONO-4538 was administered intravenously on days 1, 22 and 36 in the first 7 weeks, and then repeated every 2 weeks. Result Seventeen patients (male/female: 10/7, PS 0/1: 4/13, NSCLC: 5, melanoma: 4, CRC: 4, thymic cancer: 2, esophageal cancer: 1, thyroid cancer: 1, aged 34–74 (median: 61) years) were enrolled at a dose of 1 (n = 3), 3 (n = 5), 10 (n = 6) and 20 (n = 3) mg/kg. The main adverse reactions included lymphopenia (59%), eosinophilia (47%), pyrexia (29%), hypoalbuminemia (29%) and ventricular extrasystole (24%). Grade 3 lymphopenia was observed in two patients, which was resolved without any treatment. No dose-limiting toxic effects were observed up to the dose of 20 mg/kg. The serum concentration of ONO-4538 declined slowly with a terminal half-life of 13–21 days, and the AUC increased proportionally within the dose range investigated. Two out of 17 patients had human anti-human antibodies to ONO-4538 during treatment; however, no remarkable clinical findings were observed. Partial responses were achieved in three patients (CRC at 1 mg/kg, melanoma at 3 mg/kg and thyroid cancer at 10 mg/kg), and the latter two patients continued the treatment >1 years. Three patients (NSCLC at 3 mg/kg, thymic cancer and NSCLC at 10 mg/kg) had stable disease, and the former two patients continued the treatment >6 months. Conclusion ONO-4538 was well tolerated up to the 20 mg/kg and antitumor activity was observed in several types of advanced solid tumors.

Population pharmacokinetics of farletuzumab, a humanized monoclonal antibody against folate receptor alpha, in epithelial ovarian cancer

PurposeThe purpose of this analysis was to develop a population pharmacokinetic model for farletuzumab, a humanized immunoglobulin (Ig)G1 monoclonal antibody (mAb) to the folate receptor alpha, which is a receptor over-expressed in ovarian cancer, but largely absent from normal tissue.MethodsIn total, 2,472 samples were included in the building of the pharmacokinetic model. Farletuzumab 12.5–400 mg/m2 had been administered via intravenous infusion to 79 patients with advanced ovarian cancer enrolled in one of the two clinical studies. Data were analyzed by a nonlinear mixed-effects modeling approach.ResultsFarletuzumab pharmacokinetics was best described by a two-compartment model with first-order (linear) elimination. In the final model, estimated values of clearance and volume of distribution of the central compartment were 0.00784 l/h and 3.00 l, respectively. Body weight was the only covariate investigated that explained inter-patient variability in clearance and the central volume of distribution. There was no effect of age, human anti-human antibodies, or concomitant chemotherapy on the pharmacokinetics of farletuzumab. Simulations showed that, when the mg/kg/week dose was maintained, steady-state exposure to farletuzumab was similar with dosing every week or every 3 weeks.ConclusionsThe pharmacokinetic parameters of farletuzumab are similar to those of other IgG mAbs. The results support weight-based dosing of farletuzumab on a weekly or 3-weekly schedule.

Abstract 3519: Appropriate crosslinking is required for co-stimulatory activity of human CD27 antibody in a transgenic mouse model

Abstract CD27 is a member of TNFR superfamily. It constitutively expresses on the majority of T cell and a subset of NK cells, playing key roles in T cell activation and survival and in NK cell proliferation and cytotoxicity upon interaction with ligand CD70. Some antibodies to mouse CD27 have been reported that display agonistic and anti-tumor activities while other mAbs had less anti-tumor activity and were depleting. We hypothesized that differences in these antibodies may be due to Fc receptor engagement, as has recently been shown for the adjuvant and anti-tumor activities of agonistic mouse CD40 mAbs, which is also member of TNFR superfamily. We have developed and previously described a human anti-human CD27 antibody (1F5) and a human CD27 transgenic mouse model (hCD27-Tg) to explore the therapeutic potential of targeting CD27. In this study, we examined the effect of modifying the constant regions of the 1F5 mAb on its ability to enhance antigen specific T cell responses. With the original 1F5 hIgG1 as template, a panel of 1F5 variants was made including 1F5 mIgG1, 1F5 mIgG2a, 1F5 mIgG1D265A and 1F5 hIgG1N297S using molecular cloning techniques. All of the variants retained equal binding to hCD27 as shown by ELISA and flow cytometry studies. In addition, Biacore analysis confirmed the expected pattern of binding to human and mouse FcαRs. Co-injection of 1F5 or its variants with ovalbumin enhanced antigen-specific CD8 T cell response to different extents, as detected by SIINFEKL-specific IFNα-ELISPOT and ICS. The 1F5 mIgG1 induced the highest number of IFNα-producing CD8+ cells, whereas 1F5 mIgG2a was very weak at enhancing the CD8 T cell response. The hIgG1 version of 1F5 was intermediate in activity. Introduction of the D265A mutation that disrupts FcαRs binding into the mIgG1 eliminated the co-stimulatory function of 1F5. Similarly, the 1F5 hIgG1N297S also showed reduced activity compared to the original 1F5 hIgG1. The isotype-specific effects on our anti-hCD27 mAb are surprisingly consistent with the findings described for the agonistic anti-mCD40 mAbs, and imply that engagement of the inhibitory Fcα receptors (FcαRIIb) is driving the co-stimulatory activity in this model. Interestingly, the 1F5 hIgG1 triggered a significant T cell response, despite the lack of FcαRIIb binding by Biacore analysis. The effect of these variants on anti-tumor activity in hCD27 transgenic mice is currently being investigated. The 1F5 hIgG1 mAb (CDX-1127) is currently undergoing clinical evaluation in a phase 1 trial of patients with advanced cancers. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3519. doi:1538-7445.AM2012-3519

Abstract PR6: Phase 1b/2 trial of HER3 inhibitor U3-1287 in combination with erlotinib in advanced NSCLC patients (pts): HERALD study

Abstract Background: EGFR is frequently overexpressed in NSCLC, and while many advanced NSCLC tumors initially respond to EGFR tyrosine kinase inhibitors (TKIs), development of therapeutic resistance often follows. HER3 is a key dimerization partner of HER family members, including EGFR, and activates oncogenic signaling pathways. HER3 overexpression occurs in many solid tumors and is associated with poor prognosis in lung cancer pts. Data indicate that HER3 expression may play a role in EGFR TKI resistance, suggesting that simultaneous inhibition of HER3 and EGFR may be beneficial. U3-1287 is a fully human anti-HER3 monoclonal antibody with synergistic anticancer activity in combination with anti-EGFR inhibitors in preclinical models. The phase 1b/2 HERALD trial was initiated to investigate the combination of U3-1287 with erlotinib in the treatment of advanced NSCLC pts after failure of at least 1 prior chemotherapy. Results as of August 12, 2011 are reported here. Methods: Eligible pts had stage IIIB/IV NSCLC that progressed on ≥1 prior chemotherapy treatments and were EGFR treatment-naive. In the open-label, phase 1b portion, pts received erlotinib 150 mg/day orally and U3-1287 18 mg/kg intravenously every 3 weeks (Q3W). In the event that 18 mg/kg was not tolerated based on DLT assessment, sequential cohorts were to receive de-escalating doses of U3-1287. As no DLTs were reported, the recommended phase 2 dose is 18 mg/kg Q3W for U3-1287 in combination with 150 mg/day erlotinib. The phase 2 portion of the study is a randomized, placebo-controlled, double-blind study assessing the efficacy and safety of U3-1287 combined with erlotinib relative to erlotinib alone. It is a 3-arm study of 150 mg/day erlotinib with U3-1287 high-dose (18 mg/kg Q3W), U3-1287 low-dose (18 mg/kg loading dose followed by 9 mg/kg Q3W), or placebo. Study end points include adverse event (AE) incidence, pharmacokinetics, human antihuman antibody (HAHA) formation, tumor response, and progression-free survival (PFS). Results: The phase 1b portion of the trial enrolled 7 pts (4 male), with a median age of 68 years (range, 48–78). There were no reported DLTs. Erlotinib-related AEs reported in ≥2 pts were rash (6 pts), diarrhea (4), dry skin (3), decreased appetite (3), stomatitis (3), dehydration, dermatitis acneiform, dysgeusia, mucosal inflammation, nausea, and skin exfoliation (2 each). The only U3-1287–related AE reported in ≥2 pts was decreased appetite (2 pts). AEs grade ≥3 occurred in 2 pts: one grade 3 case each of pain, fatigue, headache, dehydration, diarrhea, and blood creatinine increase; none were related to U3-1287. Three pts had 3 serious AEs: grade 3 pain (unrelated to study treatment), grade 3 dehydration (erlotinib-related), and grade 1 decreased appetite (erlotinib- and U3-1287-related). All seven pts tested negative for HAHA formation after drug administration. As of Aug 12, 3 pts have ended study treatment due to disease progression. Four pts had best responses of stable disease lasting 86, 87, 90, and 117 days. As of Aug 12, in the phase 2 portion, 11 pts (4 male) have been screened; first study treatment was June 21. Pts had a median age of 70 years (range, 49–83). There have been 2 serious AEs in 1 pt, both unrelated to study treatment: grade 2 cardiac disorder requiring hospitalization and death due to multi-organ failure. Treatment-related AEs reported were diarrhea (3 pts), rash (3), cardiac disorder, decreased appetite, nausea, and vomiting (1 each); all were grade 1 or 2. Conclusions: Results across the phase 1b and 2 portions indicate that U3-1287 in combination with erlotinib is generally well tolerated. This abstract is also presented as Poster B38.