Adaptor-associated kinase 1 (AAK1), a member of the Ark1/Prk1 family of serine/threonine kinases, plays a role in modulating clatherin coated endocytosis of specific surface receptors. We have demonstrated that AAK1 inhibitors are efficacious in rodent models of neuropathic pain (Kostich et al., 2016). Here we have characterized the binding properties and distribution pattern of the tritiated AAK1 radioligand, [3H]BMT-046091, in rodents and cynomolgus monkeys, and used the radioligand to measure the brain target occupancy following systemic administration of AAK1 inhibitors. We have found that [3H]BMT-046091 is potent and selective AAK1 inhibitor. It inhibits AAK1 phosphorylation of a peptide derived from a physiologic substrate, the μ2 subunit of the adaptor protein complex, with an IC50 value of 2.8 nM, and is inactive at >5 μM in a panel of functional or binding assays for receptors, transporters and enzymes. [3H]BMT-046091 binding in the brain is absent in the AAK1 knockout mouse, and is displaceable with a high concentration of AAK1 inhibitors in wild type mice. Specific [3H]BMT-046091 binding is widespread in the brain and spinal cord with the highest density in the cortex, hippocampus, amygdala, striatum and thalamus. In the spinal cord, [3H]BMT-046091 binding appears enriched in the dorsal horn superficial layers. Oral administration of LP-935509, an AAK1 inhibitor, results in a dose-dependent occupation of AAK1 binding sites in the brain and spinal cord. The increase in AAK1 binding site occupancy by LP-935509 correlates with the decrease in antinociceptive responses in the rat chronic constriction injury model of neuropathic pain.

中文翻译：

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[3H] BMT-046091一种有效的和选择性的放射性配体，可确定AAK1的分布和目标参与度。

衔接子相关激酶1（AAK1），丝氨酸/苏氨酸激酶Ark1 / Prk1家族的成员，在调节特定表面受体的鞘膜包被的内吞作用中发挥作用。我们已经证明AAK1抑制剂在神经性疼痛的啮齿动物模型中有效（Kostich et al。，2016）。在这里，我们表征了the化的AAK1放射性配体[3H] BMT-046091在啮齿动物和食蟹猴中的结合特性和分布模式，并使用了放射性配体来测量全身施用AAK1抑制剂后的脑靶标占有率。我们已经发现[3H] BMT-046091是有效的和选择性的AAK1抑制剂。它抑制源自生理底物（衔接蛋白复合物的μ2亚基）的肽的AAK1磷酸化，IC5​​0值为2.8 nM，在>时无活性。5μM，用于受体，转运蛋白和酶的功能或结合测定。在AAK1基因敲除小鼠中[3H] BMT-046091在大脑中不存在结合，并且在野生型小鼠中可以用高浓度的AAK1抑制剂置换。[3H] BMT-046091特异性结合在大脑和脊髓中广泛分布，在皮质，海马，杏仁核，纹状体和丘脑中密度最高。在脊髓中，[3H] BMT-046091结合似乎富集在背角浅层。口服AAK1抑制剂LP-935509导致大脑和脊髓中AAK1结合位点的剂量依赖性占领。LP-935509增加的AAK1结合位点占有率与神经性疼痛的大鼠慢性收缩性损伤模型中抗伤害感受反应的降低有关。