Annals of Oncology Advance Access originally published online on August 9, 2007
Annals of Oncology 2007 18(10):1704-1707; doi:10.1093/annonc/mdm280
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© 2007 European Society for Medical Oncology
phase I and pharmacokinetics |
Pharmacokinetic and safety study of subcutaneously administered weekly ING-1, a human engineeredTM monoclonal antibody targeting human EpCAM, in patients with advanced solid tumors
1 Albert Einstein Cancer Center
2 Department of Oncology
3 Division of Gynecologic Oncology, Albert Einstein College of Medicine and Montefiore Medical Center, New York
4 XOMA (US) LLC, Berkeley CA, USA
* Correspondence to: Dr S. Goel and Dr S. Mani, Weiler Hospital, Department of Oncology, 1825 Eastchester Road, Bronx, New York, NY 10461, USA. Tel: +1 718-904-2488; Fax: +1 718-904-2830; E-mail: sgoel{at}montefiore.org
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Abstract |
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Background: ING-1 is a high-affinity, human engineeredTM monoclonal antibody that recognizes a 40 kilodalton epithelial cell adhesion molecule (EpCAM) glycoprotein that is expressed in high levels on most adenocarcinomas and is an attractive target for immunotherapy.
Methods: ING-1 was administered subcutaneously weekly at doses between 0.1 and 2 mg/kg/week. Pharmacokinetic samples were drawn during weeks 1 and 6.
Results: Fourteen patients with advanced refractory cancer received a median of 6 (range 1–9) doses of ING-1. At 1 mg/kg, a 62-year-old man with colon cancer developed reversible grade 3 pancreatitis after the third dose. His plasma ING-1 levels were similar to the other two patients dosed at 1 mg/kg. Two patients dosed at 0.6 mg/kg experienced stable disease at 6 weeks. Peak drug levels increased with dose and time, suggesting drug accumulation with repeated dosing. Low human anti-human antibody response was noted in three of the 13 patients assessed and was directed towards the variable region of ING-1.
Conclusions: Weekly ING-1 administered subcutaneously was well tolerated at 0.6 mg/kg/week and further experience at this dose is warranted to demonstrate safety. The risk of pancreatitis and the marginal anti-tumor effect may preclude further monotherapy studies; however, combination studies with chemotherapy are warranted.
Key words: pharmacokinetics, Phase I, ING-1, EpCAM, cancer
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introduction |
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TopAbstractintroductionpatients and methodsresultsdiscussionAcknowledgementsReferences |
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ING-1 is a high-affinity, human engineeredTM monoclonal antibody (MAb) that recognizes a 40 kilodalton glycoprotein that functions as an epithelial cell adhesion molecule (EpCAM). EpCAM is detected at the basolateral cell membrane of all simple, pseudostratified and transitional epithelia and its targeting in xenograft models of human breast and colon adenocarcinoma has resulted in prevention of tumor formation [1–3]. It may thus be an attractive target for immunotherapy [4, 5].
Monoclonal antibodies are thought to be cytotoxic to tumor cells by activating antibody-dependent cell-mediated cytotoxicity (ADCC) [6, 7], rather than complement-dependent cytotoxicity [4, 8, 9], since most tumors express increased amounts of complement-inhibiting regulators [6, 7]. ING-1 has been evaluated with several antigen-positive human tumor cell lines by two ADCC assays, and has elicited potent killing at concentrations 
1 µg/mL.
ING-1 has been studied as a single intravenous infusion every 3 weeks [10] and as weekly intravenous infusions, including 1 mg of 131I-labeled ING-1 to study biodistribution. Antibody localization to normal and tumor tissues was seen, confirming the hypothesis that an EpCAM sink contributed to a shorter than expected half-life.
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patients and methods |
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TopAbstractintroductionpatients and methodsresultsdiscussionAcknowledgementsReferences |
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Eligible patients had the following characteristics: a pathologic diagnosis of adenocarcinoma of the ovary, breast, lung, prostate, colon or rectum, refractory to standard therapy, Eastern Cooperative Oncology Group (ECOG) performance status 0–2, age
18 years and adequate organ function. All patients gave written informed consent approved by the Institutional Review Board at Montefiore Medical Center. ING-1 was supplied by XOMA (US) LLC as a single-use vial containing sterile pyrogen-free lyophilized drug. Injections were repeated weekly for 6 weeks (one cycle). All patients underwent a complete medical history and physical examination, electrocardiogram, hematology, coagulation, chemistry and urinalysis within 4 weeks of starting drug therapy and prior to each dose. Lipase and amylase were repeated at 6 and 24 hours after the first and the sixth dose. Samples for human anti-human antibody (HAHA) against ING-1 were drawn at baseline, day 22 and end of study. Imaging studies for tumor measurement were performed at baseline and at 6 weeks. Response evaluation was performed using the response evaluation criteria in solid tumors (RECIST) [11].
This was an open-label, dose-escalating study to evaluate the safety, tolerability, bioavailability, immunogenicity and pharmacokinetics of ING-1. The starting dose was 0.1 mg/kg, based on the prior experience that 0.1 mg/kg/week intravenously was safe and on mathematical simulation models that projected peak plasma levels to be 1 µg/ml at this dose.
Plasma samples for pharmacokinetic analyses were drawn pre-dose (0 hour) and at 6, 24, 48, 96 and 168 hours in the first and sixth week and prior to dosing on days 8, 15, 22 and 29. Noncompartmental analysis was performed on the plasma concentrations. The trough concentration before each dose and peak concentration (Cmax) after the first and last dose were obtained. Compartmental analysis was also performed. Data for individual subjects were fitted by unweighted nonlinear least squares analysis on the plasma concentrations of ING-1 using a bi-exponential disposition function. The curve fits yielded three primary pharmacokinetic parameters: rising (alpha) phase half-life, declining (beta) phase half-life and coefficient to the beta phase (B). The coefficient to the beta phase was set to –B. The secondary pharmacokinetic parameters, area under the curve (AUC) and clearance/F (Cl-F-subcutaneous clearance of drug), were calculated from the primary parameters [12]. Bioavailability (Bio) was calculated as Bio = 100 x (Cl-IV)/(Cl-F), where Cl-IV is the intravenous clearance obtained from prior studies [1, 10].
EpCAM expression on tumor specimens was analysed by immunohistochemical staining using an indirect peroxidase-conjugated streptavidin immunohistochemical technique. Staining was reported at the highest level of intensity observed as is standard practice, ranging from 0 to 3+.
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results |
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Fourteen patients (6 males, 8 females) with a median age of 60 (range 47–83) years, diagnosis of advanced refractory cancer (ovary—3 patients, colon—10 patients, and lung—1 patient), and ECOG performance status of 0–1 received a total of 69 weeks of therapy with a median of 6 (range 1–9) doses of ING-1. All had received prior chemotherapy and one had prior radiation therapy. The percentage of tumor cells staining 3+ for EpCAM expression by IHC analysis was between 10% and 100%.
At 1 mg/kg, a 63-year-old man with colon cancer developed a dose limiting toxicity (DLT) grade 3 pancreatitis with abdominal pain, lipase and amylase elevation after the third injection. His symptoms resolved with conservative management including fasting and use of analgesics and he was not treated again with the drug. The two remaining patients at this dose and one patient at 2 mg/kg were dose-reduced to 0.6 mg/kg; also three additional patients were enrolled at 0.6 mg/kg. Overall, non-hematologic toxicities were more common than hematological toxicities (Table 1), including grade 2 neutropenia. Grade 3 anemia occurred in a patient with ovarian cancer treated at 0.6 mg/kg who received a total of 9 doses over a period of 3 months with dose interruptions. ING-1 was generally safe and well tolerated at doses up to 0.6 mg/kg, determined to be the maximum tolerated dose.
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Table 2 shows the average trough and peak concentrations measured for the dose groups. The peak and AUC level of the patient who developed DLT-defining pancreatitis was similar to those of the other two patients at 1 mg/kg. Plasma peak and trough ING-1 levels after the first and last dose increased with increasing dose. Peak and trough ING-1 levels 1 week after the last dose were higher than 1 week after the first dose, suggesting drug accumulation.
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For some of the patients, not all of the data could be well fitted to a single disposition function throughout the entire dosing period, suggesting that the kinetics, particularly bioavailability, might have changed with repeated dosing. At the lowest doses, the apparent bioavailability changed considerably from first to last dose, but as the dose increased, the difference between the first and last dose diminished. Figure 1 shows the individual plasma concentrations plotted against time after 0.6 mg/kg/week dosing, along with the curve fits. After the first dose, drug level increased with average rising phase half-lives of 16–35 hours to peak levels by 2–4 days, and then declined with average half-lives of 60–131 hours.
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There was no objective tumor response observed; however, two (of 11 evaluable) patients had stable disease, lasting 9 (ovarian cancer) and 6 (colorectal cancer) weeks respectively. Three patients had HAHA responses (on day 57 (60.5 ng/mL), and on day 43 (85.8 ng/mL and 55.9 ng/mL)) of goat anti-human IgG standard. Results of the competition experiments suggest that these antibody responses were directed towards the variable region of ING-1.
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discussion |
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TopAbstractintroductionpatients and methodsresultsdiscussionAcknowledgementsReferences |
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Previously, ING-1 had been shown to be well tolerated at infusions up to 0.3 mg/kg every 3 weeks and in weekly infusions of 0.1 mg/kg. However, higher doses were associated with reversible acute pancreatitis that seemed peak-related [10]. A purportedly safer subcutaneous formulation was developed and tested; however, at 1 mg/kg we observed an acute reversible pancreatitis, suggesting that toxicity may be AUC- rather than peak-related. Pancreatitis has also previously been described with another high-affinity anti-EpCAM MAb, 3622W94 (Glaxo-Smith-Kline), in addition to transient and asymptomatic increases in pancreatic enzymes at lower doses [13, 14]. The reason for this toxicity is not clearly understood. It has been previously reported that EpCAM is expressed in normal pancreatic tissue [15] and has regulatory functions including morphogenesis of epithelial tissue, particularly the pancreas and the mammary gland [16]. Targeting and inhibition of this antigen on cell surfaces, especially those with a high expression such as those of the pancreatic acini, may lead to release of intracellular contents leading to auto-digestion, pancreatic epithelial damage and development of pancreatitis. Patients tolerated doses of 0.6 mg/kg (n = 3) without experiencing any major toxicity. However, further experience at this dose is necessary before one can be certain that pancreatitis will not occur at this dose level.
Three of 13 patients treated with ING-1 developed a low-level HAHA response, which was directed against the variable region of the antibody. For HAHA assays, a sensitivity of 250 ng/mL has been recommended as adequate by the American Association of Pharmaceutical Scientists, which is higher than the HAHA levels measured in the three patients [17]. There were too few subjects in each dose group to determine if the presence of HAHA in these subjects influenced their ING-1 pharmacokinetics.
With low weekly dosing, trough concentrations increased with repeated dosing at a rate that was greater than could be accounted for by the linear kinetic exponential disposition model. This was not observed with repeated intravenous dosing [10], suggesting that most probably an increase in the fraction of ING-1 absorbed from the injection site occurred with repeated dosing, perhaps due to saturation of local elimination sites with each subsequent dose. The declining phase half-lives should be interpreted with caution, as they could not be reliably determined because plasma levels were measured only up to 1 week after the last dose.
The lack of significant anti-tumor response in this Phase I study should not be taken as conclusive that this drug lacks therapeutic efficacy. Patients had been pre-treated with chemotherapy prior to study entry and were refractory to multiple regimens. While single-agent development of this drug in chemotherapy-refractory patients is not recommended, we would certainly advocate its further development in combination with cytotoxic drugs.
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Acknowledgements |
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This study was supported by a grant from XOMA (US) LLC.
This work was presented in part at the 41st annual meeting of the American Society of Clinical Oncology, Orlando, FL, 13–17 May 2005.
Received for publication March 5, 2007. Revision received March 27, 2007. Accepted for publication May 9, 2007.
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References |
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