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Preclinical Pharmacokinetics Evaluation of Anti-heparin-binding EGF-like Growth Factor (HB-EGF) Monoclonal Antibody Using Cynomolgus Monkeys via 89Zr-immuno-PET Study and the Determination of Drug Concentrations in Serum and Cerebrospinal Fluid

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Abstract

Purpose

Heparin-binding EGF-like growth factor (HB-EGF) is a member of the EGF family and is an important therapeutic target in some types of human cancers. KHK2866 is a humanized anti-HB-EGF monoclonal antibody IgG that neutralizes HB-EGF activity by inhibiting the binding of HB-EGF to its receptors. The phase I study of KHK2866 was discontinued because of neuropsychiatric toxicity. In this study, the pharmacokinetics of KHK2866 was evaluated by 89Zr-immuno-PET study and the determination of drug concentrations in serum and cerebrospinal fluid using cynomolgus monkeys was performed in order to predict neurotoxicity in a reverse-translational manner.

Methods

KHK2866 was radiolabeled with 89Zr for preclinical evaluations in normal cynomolgus monkeys and its distribution was analyzed. Furthermore, as a separate study, KHK2866 concentrations in serum and cerebrospinal fluid were determined after administration of a single dose.

Results

PET studies with monkeys revealed 89Zr-KHK2866 accumulation in the liver, spleen and joints of multiple parts, but not in brain. In addition, the pharmacokinetic analyses in serum and CSF demonstrated a low penetration of KHK2866 into the brain.

Conclusions

These studies indicate the difficulty of prediction for neuropsychiatric toxicity of monoclonal antibodies in human by means of pharmacokinetic evaluations using cynomolgus monkeys.

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Abbreviations

BBB:

Blood–brain barrier

Bq:

Becquerel

CSF:

Cerebrospinal fluid

CT:

Computed tomography

CTAC:

CT-based attenuation correction

EGF:

Epidermal growth factor

ELISA:

Enzyme-linked immunosorbent assay

FWHM:

Full width half maximum

HB-EGF:

Heparin-binding EGF-like growth factor

HPLC:

High performance liquid chromatography

IACUC:

Institutional animal care and use committee

LLOQ:

Lower limit of quantification

MIP:

Maximum intensity projections

MTD:

Maximum tolerated dose

PET:

Positron emission tomography

PK:

Pharmacokinetics

QC:

Quality control

SUV:

Standard uptake value

TLC:

Thin-layer chromatography

VOI:

Volume of interest

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ACKNOWLEDGMENTS AND DISCLOSURES

We thank Drs. T. Tawara, N. Mimura, A. Takami, Y Yoda, J Hosogi, K Fujita, H. Kaito, and TW. Poh for excellent support and helpful suggestion. This research was funded by Kyowa Hakko Kirin, Co. Ltd.

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Authors and Affiliations

  1. Singapore Translational Research Laboratory, Kyowa Hakko Kirin (Singapore) Pte. Ltd., 11, Biopolis Way, #05-08, Helios, Singapore, 138667

    Noriyuki Kasai & Kazuya Yamano

  2. R&D Division, Kyowa Hakko Kirin Co. Ltd., 1188, Shimotogari, Nagaizumi-cho, Sunto-gun, Shizuoka, 411-8731, Japan

    Maiko Adachi

Authors
  1. Noriyuki Kasai
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Corresponding author

Correspondence to Noriyuki Kasai.

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Fig. S1

Comparison of antigen-binding activities between cold Zr-KHK2866 and unlabeled KHK2866 by antigen-coated ELISA. The antigen-binding activity was compared by antigen-coated ELISA as described below briefly. Coated antigen; recombinant human HB-EGF (R&D Systems), analytes; cold Zr-KHK2866 or unlabeled KHK2866 (0.137-100 ng/ml), detection antibody; anti-human IgG (whole molecule)-peroxidase antibody produced in rabbit (Sigma), substrate; TMB (3,3′,5,5;-tetramethylbenzidine) chromogen solution (Life Technologies), microplate reader; Varioskan Flash (Thermo Scientific). OD450 nm; optical density at 450 nm. (JPEG 49 kb)

High resolution image (TIFF 20894 kb)

Fig. S2

Calibration curves of the electrochemiluminescence assay for the determination of KHK2866 and KM8047 concentrations in cynomolgus monkey serum (A) and CSF (B). Calibration curves were generated by 4-parameter regression of the natural logarithmic transformed nominal concentrations of the calibration standards (X axis), except for the 0 μg/ml sample, and the natural logarithmic transformed mean values of duplicate signal intensity (Y axis). (JPEG 175 kb)

High resolution image (TIFF 77199 kb)

Table S1

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Table S2

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Kasai, N., Adachi, M. & Yamano, K. Preclinical Pharmacokinetics Evaluation of Anti-heparin-binding EGF-like Growth Factor (HB-EGF) Monoclonal Antibody Using Cynomolgus Monkeys via 89Zr-immuno-PET Study and the Determination of Drug Concentrations in Serum and Cerebrospinal Fluid. Pharm Res 33, 476–486 (2016). https://doi.org/10.1007/s11095-015-1803-2

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  • DOI: https://doi.org/10.1007/s11095-015-1803-2

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