Preclinical evaluation of [F]D3FSP, deuterated AV-45, for imaging of β-amyloid in the brain.

INTRODUCTION

Since the approval of three F labeled β-amyloid-targeting PET imaging agents, Amyvid (florbetapir f18, AV-45), Neuraceq (florbetaben f18, AV-1) and Vizamyl (flutemetamol f18, F-PIB), they have increasingly been employed to assist differential diagnosis of Alzheimer's disease in patients with dementia. Also, they are frequently used in selecting patients participating drug trials aiming to reduce β-amyloid (Aβ) plaques in the brain. The first approved tracer in this class was [F]AV-45, which is metabolized rapidly in blood and some of its N-demethylated metabolites cross the blood brain barrier and resulted in lowering the image contrast. To improve metabolic stability of [F]AV-45, we hypothesized that substituting N-CH with N-CD at the metabolically labile position, creating [F]D3FSP, may reduce in vivo N-demethylation. We report the preclinical evaluation of [F]D3FSP as an Aβ imaging agent.

METHODS

Preclinical pharmacology of [F]D3FSP was evaluated using in vitro autoradiography and competitive binding assay. Biodistribution of [F]D3FSP was evaluated in wild-type CD-1 mice. In vivo metabolism in mice and in vitro microsomal metabolism were analyzed by HPLC. Single dose acute toxicity of D3FSP was also performed in rats.

RESULTS

[F]D3FSP showed high binding affinity to β-amyloid plaques (Ki = 3.44 ± 1.22 nM, a value similar as AV-45 (Ki = 4.02 ± 0.22 nM)), and displayed excellent β-amyloid binding in AD brain sections consistent with known Aβ regional distribution. After an iv injection it exhibited good initial brain uptake and fast washout in wild-type CD-1 mice. In vitro microsomal metabolism and in vivo metabolism in mice did not result in any significant differences between [F]D3FSP and [F]AV-45. No treatment-related mortality or any adverse effects were observed in single dose acute toxicity studies administered at hundred-folds of maximum human dose.

CONCLUSION

A new small molecule, [F]D3FSP, was prepared and tested as an alternative to [F]AV-45 to reduce N-demethylation in vivo. This strategy did not lead to better in vivo stability. However, [F]D3FSP displayed very similar Aβ targeting property comparable to [F]AV-45. Preclinical studies suggest that [F]D3FSP is useful as a β-amyloid-targeting PET imaging agent.