三环 1,2-噻嗪衍生物对脂多糖预孵育或小胶质细胞样细胞共培养诱导的神经炎症的影响。

Effect of tricyclic 1,2-thiazine derivatives in neuroinflammation induced by preincubation with lipopolysaccharide or coculturing with microglia-like cells.

机构信息

Department of Pharmacology, Faculty of Medicine, Wroclaw Medical University, ul. Mikulicza-Radeckiego 2, 50-345, Wroclaw, Poland.

Department of Inorganic Chemistry, Wroclaw Medical University, Wroclaw, Poland.

出版信息

Pharmacol Rep. 2022 Oct;74(5):890-908. doi: 10.1007/s43440-022-00414-8. Epub 2022 Sep 21.

DOI:10.1007/s43440-022-00414-8

PMID:36129673

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9584986/

Abstract

BACKGROUND

Alzheimer's disease (AD) is considered the most common cause of dementia among the elderly. One of the modifiable causes of AD is neuroinflammation. The current study aimed to investigate the influence of new tricyclic 1,2-thiazine derivatives on in vitro model of neuroinflammation and their ability to cross the blood-brain barrier (BBB).

METHODS

The potential anti-inflammatory effect of new tricyclic 1,2-thiazine derivatives (TP1, TP4, TP5, TP6, TP7, TP8, TP9, TP10) was assessed in SH-SY5Y cells differentiated to the neuron-like phenotype incubated with bacterial lipopolysaccharide (5 or 50 μg/ml) or THP-1 microglial cell culture supernatant using MTT, DCF-DA, Griess, and fast halo (FHA) assays. Additionally, for cultures preincubated with 50 µg/ml lipopolysaccharide (LPS), a cyclooxygenase (COX) activity assay was performed. Finally, the potential ability of tested compounds to cross the BBB was evaluated by computational studies. Molecular docking was performed with the TLR4/MD-2 complex to assess the possibility of binding the tested compounds in the LPS binding pocket. Prediction of ADMET parameters (absorption, distribution, metabolism, excretion and toxicity) was also conducted.

RESULTS

The unfavorable effect of LPS and co-culture with THP-1 cells on neuronal cell viability was counteracted with TP1 and TP4 in all tested concentrations. Tested compounds reduced the oxidative and nitrosative stress induced by both LPS and microglia activation and also reduced DNA damage. Furthermore, new derivatives inhibited total COX activity. Additionally, new compounds would cross the BBB with high probability and reach concentrations in the brain not lower than in the serum. The binding affinity at the TLR4/MD-2 complex binding site of TP4 and TP8 compounds is similar to that of the drug donepezil used in Alzheimer's disease. The ADMET analysis showed that the tested compounds should not be toxic and should show high intestinal absorption.

CONCLUSIONS

New tricyclic 1,2-thiazine derivatives exert a neuroregenerative effect in the neuroinflammation model, presumably via their inhibitory influence on COX activity and reduction of oxidative and nitrosative stress.

摘要

背景

阿尔茨海默病（AD）被认为是老年人中最常见的痴呆症病因。AD 的一种可改变的病因是神经炎症。本研究旨在研究新型三环 1,2-噻嗪衍生物对神经炎症体外模型的影响及其穿过血脑屏障（BBB）的能力。

方法

在用细菌脂多糖（5 或 50μg/ml）或 THP-1 小胶质细胞培养上清液孵育分化为神经元样表型的 SH-SY5Y 细胞中，使用 MTT、DCF-DA、Griess 和快速 halo（FHA）测定法评估新型三环 1,2-噻嗪衍生物（TP1、TP4、TP5、TP6、TP7、TP8、TP9、TP10）的潜在抗炎作用。此外，对于用 50μg/ml 脂多糖（LPS）预孵育的培养物，进行环氧化酶（COX）活性测定。最后，通过计算研究评估了测试化合物穿过 BBB 的潜在能力。与 TLR4/MD-2 复合物进行分子对接，以评估测试化合物在 LPS 结合口袋中结合的可能性。还进行了 ADMET 参数（吸收、分布、代谢、排泄和毒性）的预测。

结果

LPS 的不利影响和与 THP-1 细胞的共培养对神经元细胞活力的影响，用所有测试浓度的 TP1 和 TP4 得到了逆转。测试化合物降低了由 LPS 和小胶质细胞激活引起的氧化和硝化应激，并减少了 DNA 损伤。此外，新衍生物抑制了总 COX 活性。此外，新化合物很有可能穿过 BBB，并在大脑中达到的浓度不低于血清中的浓度。TP4 和 TP8 化合物在 TLR4/MD-2 复合物结合位点的结合亲和力与用于阿尔茨海默病的药物多奈哌齐相似。ADMET 分析表明，测试化合物不应有毒，并应表现出高肠道吸收。