Key Laboratory of Preclinical Study for New Drugs of Gansu Province, Research Unit of Peptide Science, Chinese Academy of Medical Sciences, 2019RU066, Institute of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, 730000, PR China.
Key Laboratory of Preclinical Study for New Drugs of Gansu Province, Research Unit of Peptide Science, Chinese Academy of Medical Sciences, 2019RU066, Institute of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, 730000, PR China.
Eur J Med Chem. 2023 Aug 5;256:115412. doi: 10.1016/j.ejmech.2023.115412. Epub 2023 Apr 28.
Isoquinoline alkaloids are a rich source of multimodal agents with distinctive structural specificity and various pharmacological activities. In the present report, we propose a combination of design, synthesis, computational study, primary in-vitro screening using the lipopolysaccharide (LPS)-induced RAW 264.7 cell line, and in-vivo evaluation in mice models as a novel approach to speed up anti-inflammatory drugs discovery. The nitric oxide (NO) inhibitory effect of new compounds revealed that all of them displayed the potent NO inhibitory ability in a dose-dependent manner with no obvious cytotoxicity. A series of the model compounds 7a, 7b, 7d, 7f, and 7g have been identified as the most promising, with IC values of 47.76 μM, 33.8 μM, 20.76 μM, 26.74 μM, and 47.8 μM respectively in LPS-induced RAW 264.7 cell line. Structure-activity relationship (SAR) studies on a range of derivatives aided in identifying key pharmacophores in the lead compound. Western blotting data of 7d identified that our synthesized compounds can down-regulate and suppress the expression of the key inflammatory enzyme, inducible nitric oxide synthase (iNOS). These results suggested that synthesized compounds may be potent anti-inflammatory agents, inhibiting the NO-release, in turn, iNOS inflammatory pathways. Furthermore, in-vivo anti-inflammatory detection via xylene-induced ear edema in mice revealed that these compounds could also inhibit swelling in mice, with model compound 7h showing an inhibition activity (64.4%) at a concentration of 10 mg/kg comparable to the reference drug celecoxib. Molecular docking results showed that shortlisted compounds (7b, 7c, 7d, 7e, and 7h) had a potential binding affinity for iNOS with low energies, with S-Score to be -7.57, -8.22, -7.35, -8.95, -9.94 kcal/mol, respectively. All results demonstrated that the newly synthesized chiral pyrazolo isoquinoline derivatives are highly potential anti-inflammatory agents.
异喹啉生物碱是一种丰富的多模态药物来源，具有独特的结构特异性和各种药理学活性。在本报告中，我们提出了一种将设计、合成、计算研究、使用脂多糖 (LPS) 诱导的 RAW 264.7 细胞系进行初步体外筛选以及在小鼠模型中进行体内评估相结合的新方法，以加速抗炎药物的发现。新化合物的一氧化氮 (NO) 抑制作用表明，所有化合物均表现出剂量依赖性的强 NO 抑制能力，且无明显细胞毒性。一系列模型化合物 7a、7b、7d、7f 和 7g 已被确定为最有前途的化合物，它们在 LPS 诱导的 RAW 264.7 细胞系中的 IC 值分别为 47.76 μM、33.8 μM、20.76 μM、26.74 μM 和 47.8 μM。对一系列衍生物的构效关系 (SAR) 研究有助于确定先导化合物中的关键药效团。7d 的 Western blot 数据表明，我们合成的化合物可以下调并抑制关键炎症酶诱导型一氧化氮合酶 (iNOS) 的表达。这些结果表明，合成的化合物可能是有效的抗炎药物，可抑制 NO 释放，从而抑制 iNOS 炎症途径。此外，通过二甲苯诱导的小鼠耳肿胀进行的体内抗炎检测表明，这些化合物还可以抑制小鼠肿胀，模型化合物 7h 在 10 mg/kg 浓度下的抑制活性（64.4%）可与参考药物塞来昔布相媲美。分子对接结果表明，筛选出的化合物（7b、7c、7d、7e 和 7h）对 iNOS 具有潜在的结合亲和力，能量较低，S-Score 分别为-7.57、-8.22、-7.35、-8.95 和-9.94 kcal/mol。所有结果均表明，新合成的手性吡唑并异喹啉衍生物具有很高的抗炎潜力。
