Bruton's tyrosine kinase (Btk) is an attractive target for the treatment of a wide array of B-cell malignancies and autoimmune diseases. Small-molecule covalent irreversible Btk inhibitors targeting Cys481 have been developed for the treatment of such diseases. In clinical trials, probe molecules are required in occupancy studies to measure the level of engagement of the protein by these covalent irreversible inhibitors. The result of this pharmacodynamic (PD) activity provides guidance for appropriate dosage selection to optimize inhibition of the drug target and correlation of target inhibition with disease treatment efficacy. This information is crucial for successful evaluation of drug candidates in clinical trials. Based on the pyridine carboxamide scaffold of a novel solvent-accessible pocket (SAP) series of covalent irreversible Btk inhibitors, we successfully developed a potent and selective affinity-based biotinylated probe 12 (2-[(4-{4-[5-(1-{5-[(3aS,4S,6aR)-2-oxo-hexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamido}-3,6,9,12-tetraoxapentadecan-15-amido)pentanoyl]piperazine-1-carbonyl}phenyl)amino]-6-[1-(prop-2-enoyl)piperidin-4-yl]pyridine-3-carboxamide). Compound 12 has been used in Btk occupancy assays for preclinical studies to determine the therapeutic efficacy of Btk inhibition in two mouse lupus models driven by TLR7 activation and type I interferon.

中文翻译：

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发现基于亲和力的Btk占位检测探针。

布鲁顿酪氨酸激酶（Btk）是治疗广泛的B细胞恶性肿瘤和自身免疫性疾病的诱人靶标。已经开发出靶向Cys481的小分子共价不可逆Btk抑制剂来治疗此类疾病。在临床试验中，占用研究需要探针分子来测量这些共价不可逆抑制剂对蛋白质的结合水平。这种药效学（PD）活性的结果为适当剂量选择提供了指导，以优化药物靶标的抑制作用以及靶标抑制作用与疾病治疗功效的相关性。该信息对于在临床试验中成功评估候选药物至关重要。基于新型共价不可逆Btk抑制剂的溶剂可及口袋（SAP）系列的吡啶羧酰胺支架，我们成功开发了一种有效的基于选择性亲和力的生物素化探针12（2-[（4- {4- [5-（ 1- {5-[（3aS，4S，6aR）-2-氧代六氢-1H-噻吩并[3,4-d]咪唑-4-基]戊酰胺} -3,6,9,12-四氧杂十五烷-15 -酰胺基）戊酰基]哌嗪-1-羰基}苯基）氨基] -6- [1-（丙-2-烯酰基）哌啶-4-基]吡啶-3-甲酰胺）。化合物12已用于临床前研究的Btk占用测定中，以确定Btk抑制在两种由TLR7激活和I型干扰素驱动的小鼠狼疮模型中的治疗功效。12-四氧杂十五烷-15-氨基）戊酰基]哌嗪-1-羰基}苯基）氨基] -6- [1-（丙-2-烯酰基）哌啶-4-基]吡啶-3-羧酰胺）。化合物12已用于临床前研究的Btk占用测定中，以确定Btk抑制在两种由TLR7激活和I型干扰素驱动的小鼠狼疮模型中的治疗功效。12-四氧杂十五烷-15-氨基）戊酰基]哌嗪-1-羰基}苯基）氨基] -6- [1-（丙-2-烯酰基）哌啶-4-基]吡啶-3-羧酰胺）。化合物12已用于临床前研究的Btk占用测定中，以确定Btk抑制在两种由TLR7激活和I型干扰素驱动的小鼠狼疮模型中的治疗功效。