(Z)-Pugnac is an O-GlcNAc-β-N-acetylglucosaminidase (O-GlcNAcase) and β-hexosaminidase inhibitor, Ki values are 46 and 36 nM respectively[1]. (Z)-PUGNAc is a stereoisomer of PUGNAc and is a more potent O-GlcNAcase inhibitor than (E)-PUGNAc both in vitro and in cells [2].
In vitro, treatment of HeLa and HEK cells with (Z)-PUGNAc (50 μM) for 48 h increased the intracellular O-GlcNAc level[2]. Treatment of Jurkat cells with (Z)-PUGNAc (100 μM) induced an increase in O-GlcNAc levels, resulting in increased cell swelling and decreased water diffusion[3]. Treatment of Min6 cells with (Z)-PUGNAc (100 μM) for 8 h upregulated the intracellular O-GlcNAc level without cytotoxicity[4].
In vivo, (Z)-PUGNAc (200 μmol/kg; i.v.) treatment of traumatic hemorrhage SD rats increased cardiac O-GlcNAc levels by approximately 2-fold, improved cardiac function and organ perfusion, and reduced circulating inflammatory cytokines[5]. (Z)-PUGNAc (7 mg/kg; i.v.) treatment of traumatic hemorrhage SD rats maintained O-GlcNAc levels in the liver and lungs, attenuated NF-κB activation in the liver, and attenuated the increase in inducible nitric oxide synthase expression[6].
References:
[1] Macauley M S, Whitworth G E, Debowski A W, et al. O-GlcNAcase Uses Substrate-assisted Catalysis: Kineticanalysis And Development Of Highly Selective Mechanism-Inspiredinhibitors[J]. Journal of Biological Chemistry, 2005, 280(27): 25313-25322.
[2] Perreira M, Kim E J, Thomas C J, et al. Inhibition of O-GlcNAcase by PUGNAc is dependent upon the oxime stereochemistry[J]. Bioorganic & medicinal chemistry, 2006, 14(3): 837-846.
[3] Nagy T, Balasa A, Frank D, et al. O-GlcNAc modification of proteins affects volume regulation in Jurkat cells[J]. European Biophysics Journal, 2010, 39: 1207-1217.
[4] Gao Y, Parker G J, Hart G W. Streptozotocin-induced β-cell death is independent of its inhibition of O-GlcNAcase in pancreatic Min6 cells[J]. Archives of biochemistry and biophysics, 2000, 383(2): 296-302.
[5] Zou L, Yang S, Hu S, et al. The protective effects of PUGNAc on cardiac function after trauma-hemorrhage are mediated via increased protein O-GlcNAc levels[J]. Shock, 2007, 27(4): 402-408.
[6] Nöt L G, Brocks C A, Vámhidy L, et al. Increased O-linked β-N-acetylglucosamine levels on proteins improves survival, reduces inflammation and organ damage 24 hours after trauma-hemorrhage in rats[J]. Critical care medicine, 2010, 38(2): 562-571.
(Z)-PUGNAc是一种O-GlcNAc-β-N-乙酰氨基葡萄糖苷酶（O-GlcNAcase）和β-氨基己糖苷酶（β-Hexosaminidase）抑制剂，Ki值分别为46和36nM[1]。(Z)-PUGNAc是PUGNAc 的立体异构体，无论在体外还是在细胞内，它都是比(E)-PUGNAc更有效的O-GlcNAcase 抑制剂[2]。
在体外，(Z)-PUGNAc（50μM）处理HeLa和HEK细胞48 h，能够增强细胞内O-GlcNAc的水平[2]。(Z)-PUGNAc（100μM）处理Jurkat细胞，诱导了O-GlcNAc水平升高，引起了细胞肿胀增加和水扩散减少[3]。(Z)-PUGNAc（100μM）处理Min6细胞8 h，上调了细胞内O-GlcNAc 的水平，且没有细胞毒性[4]。
在体内，(Z)-PUGNAc（200 μmol/kg; i.v.）处理创伤出血SD大鼠，使心脏O-GlcNAc水平增加了约2倍，改善心脏功能和器官灌注，并减少循环炎症细胞因子[5]。(Z)-PUGNAc（7 mg/kg; i.v.）处理创伤出血SD大鼠，维持了肝脏和肺中的O-GlcNAc水平，减弱了肝脏中NF-κB的激活，减弱了诱导型一氧化氮合酶表达的增加[6]。