SPD304 is a selective inhibitor of tumor necrosis factor α (TNF-α), with an IC50 value of 22μM[1]. SPD304 promotes dissociation of TNF trimers and therefore blocks the interaction of TNF and the related receptor [2]. SPD304 has been widely used to block the TNF-α/NF-κB/CUL4B pathway to induce cell cycle arrest in S phase [3].
In vitro, SPD304 treatment at 30µM for 24h significantly induced HEK293T cell death[4]. Treatment with 25µM SPD304 for 24h significantly inhibited TNFα activity and reduced L929 cell viability[5]. Treatment of GH3 cells with 8µM SPD304 for 48h inhibited TNFα-induced upregulation of MAPK pathway proteins and inhibited cell proliferation, migration, and invasion[6]. Preincubation with 1µM SPD304 for 24 hours significantly inhibited IL-8 and IL-6 levels in human dermal fibroblasts (HDFs)[7].
In vivo, SPD304 treatment (5µg; dissolved in 10µl artificial cerebrospinal fluid containing 5% DMSO) via intracerebroventricular injection for 5min inhibited the increase in blood pressure, heart rate, and renal sympathetic activity induced by TACE (TNF-α–converting enzyme) in heart failure rats[8].
References:
[1] He M M, Smith A S, Oslob J D, et al. Small-molecule inhibition of TNF-α[J]. Science, 2005, 310(5750): 1022-1025.
[2] Alexiou P, Papakyriakou A, Ntougkos E, et al. Rationally designed less toxic SPD‐304 analogs and preliminary evaluation of their TNF inhibitory effects[J]. Archiv der Pharmazie, 2014, 347(11): 798-805.
[3] Zhang C, Chen B, Jiang K, et al. Activation of TNF‐α/NF‐κB axis enhances CRL 4 BDCAF 11 E3 ligase activity and regulates cell cycle progression in human osteosarcoma cells[J]. Molecular oncology, 2018, 12(4): 476-494.
[4] Shen Q, Zhang C, Liu H, et al. De novo design of helical peptides to inhibit tumor necrosis factor-α by disrupting its trimer formation[J]. MedChemComm, 2016, 7(4): 725-729.
[5] Mascret A, Mouhsine H, Attia G, et al. New contributions to the drug profile of TNFα inhibitor SPD304: Affinity, selectivity and ADMET considerations[J]. European Journal of Pharmacology, 2021, 907: 174285.
[6] Wu X, Gong L, Li B, et al. TNF‐α can promote membrane invasion by activating the MAPK/MMP9 signaling pathway through autocrine in bone‐invasive pituitary adenoma[J]. CNS Neuroscience & Therapeutics, 2024, 30(5): e14749.
[7] Javaid N, Patra M C, Cho D E, et al. An orally active, small-molecule TNF inhibitor that disrupts the homotrimerization interface improves inflammatory arthritis in mice[J]. Science Signaling, 2022, 15(759): eabi8713.
[8] Yu Y, Cao Y, Bell B, et al. Brain TACE (tumor necrosis factor-α–converting enzyme) contributes to sympathetic excitation in heart failure rats[J]. Hypertension, 2019, 74(1): 63-72.
SPD304是一种选择性的肿瘤坏死因子α（TNF-α）抑制剂，IC50值为22μM[1]。SPD304可促进TNF三聚体的解离，从而阻断TNF与相关受体的相互作用[2]。SPD304已被广泛用于阻断TNF-α/NF-κB/CUL4B通路，以诱导细胞周期阻滞于S期[3]。
在体外，使用30μM的SPD304处理HEK293T细胞24小时，能显著诱导细胞死亡[4]。使用25μM的SPD304处理24小时，能显著抑制TNFα活性并降低L929细胞活力[5]。用8μM的SPD304处理GH3细胞48小时，可抑制TNFα诱导的MAPK通路蛋白上调，并抑制细胞增殖、迁移和侵袭[6]。用1μM的SPD304预孵育人真皮成纤维细胞（HDFs）24小时，能显著抑制IL-8和IL-6的水平[7]。
在体内，通过脑室内注射SPD304（5μg；溶于含5%DMSO的10μl人工脑脊液）5分钟，可抑制心力衰竭大鼠中TACE(TNF-α-converting enzyme)诱导的血压、心率和肾交感神经活性的升高[6]。