Yulia Rozen

Potential 18F-labeled biomarkers for epidermal growth factor receptor tyrosine kinase

Abstract As PET candidate tracers for EGFr-TK, five 4-(anilino)quinazoline derivatives, each fluorinated in the aniline moiety, were prepared. Each was tested in vitro for inhibition of EGFr autophosphorylation in A431 cell line. The leading compounds were then radiolabeled with 18 F and cell binding experiments, biodistribution and PET studies in A431 tumor-bearing mice were performed. Metabolic studies were carried out in a mice control group. From our results, we concluded that while in vitro experiments indicates efficacy of 4-(anilino)quinazoline compounds, kinetic factors and rapid blood clearance make them unsuitable as tracers for nuclear medicine imaging of EGFr-TK.

Labeled EGFr-TK irreversible inhibitor (ML03): In vitro and in vivo properties, potential as PET biomarker for cancer and feasibility as anticancer drug

Radiosynthesis of ML03 (N‐{4‐[(4,5‐dichloro‐2‐fluorophenyl)amino]quinazolin‐6‐yl}acrylamide), an irreversible EGFr‐TK inhibitor, was developed. Its in vitro and in vivo properties, its potential as PET biomarker in cancer and the feasibility of this type of compounds to be used as anticancer drug agents were evaluated. The compound was labeled with carbon‐11 at the acryloyl amide group, via automated method with high yield, chemical and radiochemical purities. ELISA carried out with A431 lysate showed high potency of ML03 with an apparent IC50 of 0.037 nM. The irreversible binding nature of ML03 was studied and 97.5% EGFr‐TK autophosphorylation inhibition was observed in intact A431 cells 8 hr post incubation with the inhibitor. Specific binding (67%) of [11C]ML03 was obtained in cells. An A431 tumor‐bearing rat model was developed and the validity of the model was tested. In biodistribution studies carried out with tumor‐bearing rats, moderate uptake was observed in tumor and high uptake in liver, kidney and intestine. In metabolic studies, fast degradation of [11C]ML03 was observed in liver and blood indicating a short half‐life of the compound in the body. PET scan with tumor‐bearing rats confirmed the results obtained in the ex vivo biodistribution studies. Although in vitro experiments may indicate efficacy of ML03, non‐specific binding, ligand delivery and degradation in vivo make ML03 ineffective as PET bioprobe. Derivatives of ML03 with lower metabolic clearance rate and higher bioavailability should be synthesized and their potential as anticancer drugs and PET bioprobes evaluated.

Radiosynthesis of ML03, a novel positron emission tomography biomarker for targeting epidermal growth factor receptor via the labeling synthon: [11C]acryloyl chloride

An automated procedure for the radiosynthesis of the labeling synthon [11C]acryloyl chloride was developed and applied for labeling several N-acryl amides with carbon-11. [11C]-6-acrylamido-4-(3,4-dichloro-6-fluoroanilino)quinazoline (ML03), a novel PET biomarker targeting the epidermal growth factor receptor tyrosine kinase (EGFr-TK) in cancer, was successfully prepared using this labeled synthon in a fully automated manner. Two other potential anticancer drugs were also labeled using the developed methodology. The potency of ML03 to inhibit autophosphorylation of EGFr-TK was evaluated by an ELISA assay indicating a low IC(50) of 0.037nM.

Improved method for the quality assurance of (C-11)choline

[C-11]choline is a very promising radiomarker for the diagnosis of various human tumors using Positron Emission Tomography (PET). The existing quality control process for [C-11]choline is complicated and combines two HPLC methods with limited separation and sensitivity which prevent the accurate determination of the specific activity. We have developed a new efficient single HPLC method for the detection of choline chloride and dimethylaminoethanol with high resolution and sensitivity using cation-exchange chromatography.