Notch信号通路小分子抑制剂DAPT的 11 C标记及在正常兔体内的初步动态显像研究

Abstract

Objective To label N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester(DAPT), an inhibitor of the Notch signaling pathway, with 11C and perform preliminarily dynamic imaging in normal rabbit. Methods Proliferation of human pancreatic cancer cell line MiaPaCa-2 were assessed by cell counting kit-8 method after treatment with various concentrations of DAPT and CH3-DAPT. Half-maximal inhibitory concentration(IC50) was calculated. DAPT was then used as a precursor to prepare 11C-N-methyl-DAPT(11C-DAPT) with a fully automatic synthesizer. The final product was purified through semipreparative high performance liquid chromatography(HPLC). After intravenous injection of 125.8 MBq(3.4 mCi) 11C-DAPT, a normal New Zealand rabbit was subjected to dynamic whole-body PET/CT scanning. The dynamic changes in radioactivity were measured by drawing regions of interest over different organs. Results DAPT and CH3-DAPT concentration-dependently inhibited the growth of the human pancreatic cancer cell line MiaPaCa-2. The IC50 values were 64.2 and 180.0 μmol/L at 72 h after administration, respectively. 11C-DAPT was synthesized for approximately 30 min. The uncorrected radiochemical yield was 25% –35%. Radiochemical purity was above 95%. 11C-DAPT was mainly excreted through the kidney, the highest uptake was in the kidneys, and the uptake in the liver, intestine, lung, and brain was relatively low. The tracer uptake in the kidneys and liver peaked at 7 min after injection and decreased to >50% at 28 min. Conclusions 11C-DAPT can be easily and rapidly synthesized with high radiochemical purity. Preliminary PET/CT imaging can lay a foundation for further investigating 11C-DAPT as a novel molecular probe. Key words: Isotope labeling; Positron emission tomography computed tomography; Notch signaling pathway; γ-secretase inhibitor; N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butylester; Molecular probes