Hypaphorine

Hypaphorine, an indole alkaloid extracted from leguminous plants, exerts anti-inflammatory, antioxidant, and neuroprotective effects by modulating multiple signaling pathways, including PI3K/Akt/mTOR and p38/JNK. Hypaphorine is frequently investigated in studies related to neurodegenerative diseases and metabolic disorders^[1-2]. Additionally, Hypaphorine has been shown to effectively prevent osteoporosis^[3], and exhibits antiviral activity against Dengue virus^[4].

In vitro, pretreatment of human microvascular endothelial cells (HMEC-1) with Hypaphorine (0–100μM) for 6 hours, followed by lipopolysaccharide (LPS, 500ng/mL) stimulation for 48 hours, significantly suppressed LPS-induced expression of pro-inflammatory cytokines (TNF-α, IL-1β, MCP-1, VCAM-1). Hypaphorine also reversed LPS-induced upregulation of TLR4 protein and mRNA levels and downregulation of PPAR-γ. Furthermore, it blocked LPS-activated PI3K/Akt/mTOR signaling^[5]. In RAW 264.7 macrophages, Hypaphorine (12.5–50μM) pretreatment for 1 hour, followed by LPS (1μg/mL) stimulation for 24 hours, markedly inhibited LPS-induced pro-inflammatory cytokine (TNF-α, IL-1β, IL-6, IL-10, MCP-1) expression and NO/PGE₂ release, while downregulating COX-2 and iNOS protein expression. Hypaphorine also suppressed LPS-induced phosphorylation of ERK, IκBα, IKKβ, and NF-κB, and inhibited NF-κB nuclear translocation^[6].

In vivo, Hypaphorine (10mg/kg; intraperitoneal injection) administered 2 hours after LPS (20mg/kg) significantly attenuated LPS-induced acute lung injury (ALI) in male Sprague-Dawley (SD) rats. Hypaphorine reduced lung histopathological damage, decreased total cell counts and neutrophil proportions in bronchoalveolar lavage fluid (BALF), and lowered lung wet/dry weight ratio, microvascular permeability, and lactate dehydrogenase (LDH)/myeloperoxidase (MPO) activity. Mechanistically, Hypaphorine upregulated DUSP1 expression and inhibited p38/JNK activation, thereby suppressing pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-18) ^[7]. Hypaphorine (10mg/kg; intraperitoneal injection) was found to selectively inhibit acetylcholinesterase (AChE) activity across all brain regions (cortex, cerebellum, striatum, hippocampus) in normal rat after 24 hours, with the D-enantiomer demonstrating potent inhibition in key brain areas^[8].

References:
[1] Qin P, Li Y, Su Y, et al. Bifidobacterium adolescentis-derived hypaphorine alleviates acetaminophen hepatotoxicity by promoting hepatic Cry1 expression. J Transl Med. 2024 May 31;22(1):525.
[2] Chen H, Guo T, Wang D, et al. Vaccaria hypaphorine impairs RANKL-induced osteoclastogenesis by inhibition of ERK, p38, JNK and NF-κB pathway and prevents inflammatory bone loss in mice. Biomed Pharmacother. 2018 Jan;97:1155-1163.
[3] Zhang T, Yan Y, Xue Y, et al. Pharmacokinetic Study of Hypaphorine, a Potential Agent for Treating Osteoclast-based Bone Loss, on Rats Using LC-MS/MS. Comb Chem High Throughput Screen. 2022;25(11):1889-1896.
[4] Bai S, Liang H, Jiang W, et al. Identifying hypaphorine as a novel antiviral compound against dengue virus. Antiviral Res. 2025 Jun 24;240:106220.
[5] Sun H, Zhu X, Cai W, et al. Hypaphorine Attenuates Lipopolysaccharide-Induced Endothelial Inflammation via Regulation of TLR4 and PPAR-γ Dependent on PI3K/Akt/mTOR Signal Pathway. Int J Mol Sci. 2017 Apr 17;18(4):844.
[6] Sun H, Cai W, Wang X, et al. Vaccaria hypaphorine alleviates lipopolysaccharide-induced inflammation via inactivation of NFκB and ERK pathways in Raw 264.7 cells. BMC Complement Altern Med. 2017 Feb 20;17(1):120.
[7] Ding YH, Miao RX, Zhang Q. Hypaphorine exerts anti-inflammatory effects in sepsis induced acute lung injury via modulating DUSP1/p38/JNK pathway. Kaohsiung J Med Sci. 2021 Oct;37(10):883-893.
[8] Yonekawa MKA, Penteado BB, Dal'Ongaro Rodrigues A, et al. l-Hypaphorine and d-hypaphorine: Specific antiacetylcholinesterase activity in rat brain tissue. Bioorg Med Chem Lett. 2021 Sep 1;47:128206.

Hypaphorine是一种从豆科植物中提取的吲哚类生物碱，通过调控多个信号通路，PI3K/Akt/mTOR、p38/JNK等，发挥抗炎、抗氧化和神经保护的作用，常被用于神经退行性疾病和代谢紊乱相关的研究^[1-2]。同时，Hypaphorine被认为可有效预防骨质疏松症^[3]，Hypaphorine也被证明具有针对登革热病毒的抗病毒活性^[4]。

在体外，Hypaphorine（0–100μM）预处理人微血管内皮细胞（HMEC-1）6小时后，再用脂多糖（LPS，500ng/mL）刺激48小时，显著抑制了LPS诱导的促炎因子（TNF-α、IL-1β、MCP-1、VCAM-1）表达，同时逆转了LPS导致的TLR4蛋白和mRNA水平升高及PPAR-γ水平降低。此外，Hypaphorine阻断了LPS激活的PI3K/Akt/mTOR信号通路 ^[5]。Hypaphorine（12.5–50μM）预处理RAW 264.7巨噬细胞1小时后，再用脂多糖（LPS，1μg/ml）刺激24小时，显著抑制了LPS诱导的促炎因子（TNF-α、IL-1β、IL-6、IL-10、MCP-1）表达和NO、PGE2释放，同时下调COX-2和iNOS蛋白表达。此外，Hypaphorine阻断了LPS激活的ERK、IκBα、IKKβ和NF-κB磷酸化，并抑制了NF-κB核转位 ^[6]。

在体内，Hypaphorine（10mg/kg）通过腹腔注射给药，用于治疗脂多糖（LPS，20mg/kg）诱导的脓毒症急性肺损伤（ALI）雄性Sprague-Dawley（SD）大鼠模型，给药时间为LPS注射后2小时。Hypaphorine显著减轻LPS诱导的肺组织病理损伤，减少支气管肺泡灌洗液（BALF）中总细胞数和中性粒细胞比例，降低肺湿/干重比、肺微血管通透性及乳酸脱氢酶（LDH）和髓过氧化物酶（MPO）活性。此外，Hypaphorine通过上调DUSP1表达并抑制p38/JNK信号通路激活，显著降低促炎细胞因子（TNF-α、IL-6、IL-1β、IL-18）水平^[7]。Hypaphorine（10mg/kg）通过腹腔注射于正常大鼠中，24小时后检测发现，Hypaphorine在所有脑区（皮层、小脑、纹状体、海马）均表现出选择性抑制AChE活性，尤其D-对映体在关键脑区表现出强效抑制作用^[8]。