Design, Synthesis and Evaluation of the First 2-Alkynyl(aza)indole 18F Probe Targeting α-Synuclein Aggregates

<b>Background/Objectives</b>: The role of α-synuclein (α-syn) in the pathogenesis of Parkinson's disease (PD) or neurodegenerative diseases such as Lewy body dementia (LBD) and multiple system atrophy (MSA) is commonly accepted. Through different physiological dysfunctions, abnormal forms of α-syn are generated. These abnormal aggregates accumulate and alter pre- and postsynaptic transmission, in particular that of dopamine. Thus, the development of a diagnostic biomarker of synucleinopathies remains crucial and challenging. The development of an α-syn positron emission tomography (PET) radiopharmaceutical may be suitable to early diagnose and stratify patients, follow up disease progression, and evaluate future therapies. <b>Methods</b>: To develop a selective α-syn PET tracer, we synthesized an original series based on alkynyl(aza)indoles. Fifteen final ligands were synthesized bearing indoles or azaindoles from one side of the alkyne and a substituted phenyl ring for the opposite side of the alkyne. The final ligands were tested to determine Ki and/or Kd toward α-syn, tau, and Aβ. <b>Results</b>: The SAR showed that the indole series exhibited moderate to low affinity for α-syn and, moreover, lower Ki toward Aβ and tau (i.e., compound <b>39</b>, Ki_(αsyn) 21.7 nM, Ki_(Aβ) 64.4 nM, Ki_(Tau) 27.6 nM), highlighting the low potency of these series to afford an α-syn tracer. The introduction of a nitrogen on the different positions of the phenyl to obtain the corresponding azaindoles resulted for most of the compounds in better affinity for α-syn and selectivity towards Aβ compared to the indole analogs (i.e., compound <b>43</b>, Ki_(αsyn) 4.7 nM, Ki_(Aβ) 24.4 nM, and Ki_(Tau) 4.61 nM). A fluorinated azaindole derivative was prepared with a view to obtaining a ¹⁸F tracer and exhibited the highest affinity for α-syn but without selectivity against tau and Aβ. The radiosynthesis of [¹⁸F]<b>45</b> was performed in a two-step procedure starting from the tosylated and protected precursor. [¹⁸F]<b>45</b> was obtained in 85 ± 5 min with a radiochemical yield of 32 ± 3%. Molar activity, determined from a calibration with stable <b>45</b>, was around 130 GBq/µmole. The dynamic PET imaging showed that [¹⁸F]<b>45</b> was able to cross the blood-brain barrier, but non-specific uptake was observed, confirming the in vitro results. <b>Conclusions</b>: Although promising nanomolar affinity for the target, the new tracer showed mainly non-specific in vivo uptake in the rat brain, indicating that further pharmacomodulations on the azaindole series are required.