Thoracic Diseases Research Unit, Departments of Medicine and Biochemistry, Mayo Clinic, 8-23 Stabile, Rochester, MN, 55905, USA.
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, 55905, USA.
Drugs R D. 2022 Jun;22(2):165-173. doi: 10.1007/s40268-022-00389-0. Epub 2022 Apr 29.
The caspase recruitment domain-containing protein 9 (CARD9) inhibitor BRD5529 has been shown to be an effective in vitro inhibitor of Pneumocystis β-glucan-induced proinflammatory signaling, suggesting its viability as a candidate for preliminary anti-Pneumocystis drug testing in the rodent Pneumocystis pneumonia (PCP) model.
Mice were injected intraperitoneally (IP) daily with either vehicle or BRD5529 at 0.1 or 1.0 mg/kg for 2 weeks. Mouse weights were taken daily. At day 14, mice were euthanized, weighed, and analyzed by flexiVent™ for lung stiffness. Lungs, liver, and kidney were then harvested for hematoxylin and eosin (H&E) staining and pathology scoring. Lung samples were further analyzed for proinflammatory cytokines via enzyme-linked immunosorbent assay (ELISA) and extracellular matrix generation via quantitative polymerase chain reaction (qPCR). Blood collection postmortem was performed for blood chemistry analysis. Furthermore, administration of BRD5529 prior to the intratracheal inoculation of fungal β-glucans, which are known proinflammatory mediators via the Dectin-1-CARD9 pathway, resulted in significant reductions in lung tissue interleukin-6 and tumor necrosis factor-α, suggesting the exciting possibility of the use of this CARD9 inhibitor as an additional therapeutic tool in fungal infections.
BRD5529 at both IP doses resulted in no significant changes in daily or final weight gain, and analysis of lung stiffness by flexiVent™ showed no significant differences between the groups. Furthermore, ELISA results of proinflammatory cytokines showed no major differences in the respective groups. qPCR analysis of extracellular matrix transcripts were statistically similar. Examination and pathology scoring of H&E slides from lung, liver, and kidney in all groups, as well as subsequent pathology scoring, showed no significant change. Blood chemistry analysis revealed similar, non-significant patterns.
In our initial general safety and toxicology assessments, BRD5529 displayed no inherent safety concerns in the analyzed parameters. These data support broader in vivo testing of the inhibitor as a timed adjunct therapy to the deleterious proinflammatory host immune response often associated with anti-Pneumocystis therapy.
半胱天冬酶募集结构域蛋白 9（CARD9）抑制剂 BRD5529 已被证明可有效抑制卡氏肺孢子菌β-葡聚糖诱导的促炎信号，这表明它有作为候选药物进行初步抗卡氏肺孢子菌药物测试的潜力，可用于啮齿动物卡氏肺孢子菌肺炎（PCP）模型。
将小鼠每天通过腹腔内（IP）注射给予载体或 BRD5529，剂量为 0.1 或 1.0mg/kg，持续 2 周。每天记录小鼠体重。第 14 天，处死小鼠，称重，并通过 flexiVent 进行肺硬度分析。然后采集肺、肝和肾组织进行苏木精和伊红（H&E）染色和病理评分。进一步通过酶联免疫吸附试验（ELISA）分析肺组织中的促炎细胞因子，通过定量聚合酶链反应（qPCR）分析细胞外基质生成。死后采集血液进行血液化学分析。此外，在气管内接种真菌β-葡聚糖（已知通过 Dectin-1-CARD9 途径是促炎介质）之前给予 BRD5529，可显著降低肺组织中白细胞介素-6 和肿瘤坏死因子-α，这表明该 CARD9 抑制剂作为真菌感染的附加治疗工具具有令人兴奋的可能性。
BRD5529 在 IP 两种剂量下均未导致每日或最终体重增加的显著变化，flexiVent 进行的肺硬度分析显示各组间无显著差异。此外，各组间促炎细胞因子的 ELISA 结果无明显差异。细胞外基质转录物的 qPCR 分析结果也具有统计学意义。对各组肺、肝和肾的 H&E 幻灯片进行检查和病理评分，以及随后的病理评分，均未发现明显变化。血液化学分析显示相似的非显著模式。
在我们对初始安全性和毒理学的评估中，BRD5529 在分析参数中未显示出固有安全性问题。这些数据支持更广泛的体内抑制剂测试，作为抗卡氏肺孢子菌治疗中常与宿主有害炎症反应相关的定时辅助治疗。
