Abstract
In this study, we found that exposure of 20 μM α-pyrrolidinooctanophenone (α-POP), a new synthetic cathinone, time- and dose-dependently reduced viability of human aortic endothelial (HAE) cells, and the 50% lethal concentration (LC50) for α-POP in its 48-h treatment was 15.6 μM. In addition, the LC50 comparison of α-POP and α-pyrrolidinoheptanophenone (α-PHPP) toxicity against ten human cells exhibited that vascular (HAE and human aortic smooth muscle) and bronchial epithelial BEAS-2B cells were more susceptible to the cytotoxicity than neuronal (SK-N-SH and A172), gastrointestinal (DLD1 and MKN45), hepatic HepG2, renal HEK293 and pulmonary A549 cells. The results suggest that abuse of the lipophilic α-pyrrolidinophenones (PPs), such as α-POP and α-PHPP, is more likely to cause damage to the vascular, respiratory and central nervous systems. Structure-activity relationship study of 18 PPs with different alkyl chain lengths and substituents revealed that the endothelial cell toxicity depends on the alkyl chain length (α-POP > α-PHPP > PPs with shorter chains), and the presence of 4′-fluoro or 3′,4′-methylenedioxy group on α-POP and α-PHPP increased the cytotoxicity. In order to understand the cytotoxic mechanism of α-POP and F-α-POP that showed the most potent toxicity, the contribution of reactive oxygen species (ROS) production, caspase-3 activation and DNA fragmentation were investigated. The treatment of HAE cells with α-POP or F-α-POP resulted in remarkable ROS production, and the ROS production and apoptotic events were significantly prevented by pretreating the cells with an antioxidant N-acetyl-l-cysteine, suggesting that ROS-dependent signaling is primarily responsible for endothelial cell apoptosis elicited by the lipophilic synthetic cathinones.
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References
Zawilska JB, Wojcieszak J (2013) Designer cathinones—an emerging class of novel recreational drugs. Forensic Sci Int 231:42–53
Kelly JP (2011) Cathinone derivatives: a review of their chemistry, pharmacology and toxicology. Drug Test Anal 3:439–453
Nagai F, Nonaka R, Satoh H, Kamimura K (2007) The effects of non-medically used psychoactive drugs on monoamine neurotransmission in rat brain. Eur J Pharmacol 559:132–137
Cozzi NV, Foley KF (2003) Methcathinone is a substrate for the serotonin uptake transporter. Pharmacol Toxicol 93:219–225
Nencini P, Amiconi G, Befani O, Abdullahi MA, Anania MC (1984) Possible involvement of amine oxidase inhibition in the sympathetic activation induced by khat (Catha edulis) chewing in humans. J Ethnopharmacol 11:79–86
Karch SB (2015) Cathinone neurotoxicity (“The “3Ms”). Curr Neuropharmacol 13:21–25
Wood DM, Davies S, Greene SL, Button J, Holt DW, Ramsey J, Dargan PI (2010) Case series of individuals with analytically confirmed acute mephedrone toxicity. Clin Toxicol 48:924–927
Nicholson PJ, Quinn MJ, Dodd JD (2010) Headshop heartache: acute mephedrone ‘meow’ myocarditis. Heart 96:2051–2052
Maan ZN, D’Souza AR (2012) Spontaneous subcutaneous emphysema associated with mephedrone usage. Ann R Coll Surg Engl 94:e38–e40
Usui S, Matsunaga T, Ukai S, Kiho T (1997) Growth suppressing activity for endothelial cells induced from macrophages by carboxymethylated curdlan. Biosci Biotechnol Biochem 61:1924–1925
Morikawa Y, Shibata A, Okumura N, Ikari A, Sasajima Y, Suenami K, Sato K, Takekoshi Y, El-Kabbani O, Matsunaga T (2016) Sibutramine provokes apoptosis of aortic endothelial cells through altered production of reactive oxygen and nitrogen species. Toxicol Appl Pharmacol 314:1–11
Matsunaga T, Kotamraju S, Kalivendi SV, Dhanasekaran A, Joseph J, Kalyanaraman B (2004) Ceramide-induced intracellular oxidant formation, iron signaling, and apoptosis in endothelial cells: protective role of endogenous nitric oxide. J Biol Chem 279:28614–28624
Hasegawa K, Wurita A, Minakata K, Gonmori K, Nozawa H, Yamagishi I, Suzuki O, Watanabe K (2014) Identification and quantitation of a new cathinone designer drug PV9 in an “aroma liquid” product, antemortem whole blood and urine specimens, and a postmortem whole blood specimen in a fatal poisoning case. Forensic Toxicol 32:243–250
Kudo K, Usumoto Y, Kikura-Hanajiri R, Sameshima N, Tsuji A, Ikeda N (2015) A fatal case of poisoning related to new cathinone designer drugs, 4-methoxy PV8, PV9, and 4-methoxy PV9, and a dissociative agent, diphenidine. Leg Med 17:421–426
Wojcieszak J, Andrzejczak D, Woldan-Tambor A, Zawilska JB (2016) Cytotoxic activity of pyrovalerone derivatives, an emerging group of psychostimulant designer cathinones. Neurotox Res 30:239–250
den Hollander B, Sundström M, Pelander A, Ojanperä I, Mervaala E, Korpi ER, Kankuri E (2014) Keto amphetamine toxicity—focus on the redox reactivity of the cathinone designer drug mephedrone. Toxicol Sci 141:120–131
den Hollander B, Sundström M, Pelander A, Siltanen A, Ojanperä I, Mervaala E, Korpi ER, Kankuri E (2015) Mitochondrial respiratory dysfunction due to the conversion of substituted cathinones to methylbenzamides in SH-SY5Y cells. Sci Rep 5:14924. doi:10.1038/srep14924
Valente MJ, Lourdes Bastos M, Fernandes E, Carvalho F, Guedes de Pinho P, Carvalho M (2017) Neurotoxicity of β-keto amphetamines: deathly mechanisms elicited by methylone and MDPV in human dopaminergic SH-SY5Y cells. ACS Chem Neurosci. doi:10.1021/acschemneuro.6b00421
Silva B, Fernandes C, Tiritan ME, Pinto MMM, Valente MJ, Carvalho M, Gusdes de Pinho P, Remião F (2016) Chiral enantioresolution of cathinone derivatives present in “legal highs”, and enantioselectivity evaluation on cytotoxicity of 3,4-methylenedioxypyrovalerone (MDPV). Forensic Toxicol 34:372–385
Valente MJ, Araújo AM, Silva R, de Lourdes Bastos M, Carvalho F, Guedes de Pinho P, Carvalho M (2016) 3,4-Methylenedioxypyrovalerone (MDPV): in vitro mechanisms of hepatotoxicity under normothermic and hyperthermic conditions. Arch Toxicol 90:1959–1973
Paillet-Loilier M, Cesbron A, Le Boisselier R, Bourgine J, Debruyne D (2014) Emerging drugs of abuse: current perspectives on substituted cathinones. Subst Abus Rehabil 5:37–52
Springer D, Fritschi G, Maurer HH (2003) Metabolism of the new designer drug α-pyrrolidinopropiophenone (PPP) and the toxicological detection of PPP and 4′-methyl-α-pyrrolidinopropiophenone (MPPP) studied in rat urine using gas chromatography-mass spectrometry. J Chromatogr B 796:253–266
Springer D, Fritschi G, Maurer HH (2003) Metabolism and toxicological detection of the new designer drug 4′-methoxy-α-pyrrolidinopropiophenone studied in rat urine using gas chromatography–mass spectrometry. J Chromatogr B 793:331–342
Springer D, Fritschi G, Maurer HH (2003) Metabolism and toxicological detection of the new designer drug 3′,4′-methylenedioxy-α-pyrrolidinopropiophenone studied in urine using gas chromatography–mass spectrometry. J Chromatogr B 793:377–388
Westphal F, Rösner P, Junge T (2010) Differentiation of regioisomeric ring-substituted fluorophenethylamines with product ion spectrometry. Forensic Sci Int 194:53–59
Lukandu OM, Costea DE, Neppelberg E, Johannessen AC, Vintermyr OK (2008) Khat (Catha edulis) induces reactive oxygen species and apoptosis in normal human oral keratinocytes and fibroblasts. Toxicol Sci 103:311–324
Dhar-Mascareño M, Cárcamo JM, Golde DW (2005) Hypoxia–reoxygenation-induced mitochondrial damage and apoptosis in human endothelial cells are inhibited by vitamin C. Free Radic Biol Med 38:1311–1322
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We thank Dr. Akira Hara for insightful discussion and critical reading of the manuscript.
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Matsunaga, T., Morikawa, Y., Tanigawa, M. et al. Structure-activity relationship for toxicity of α-pyrrolidinophenones in human aortic endothelial cells. Forensic Toxicol 35, 309–316 (2017). https://doi.org/10.1007/s11419-017-0359-8
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DOI: https://doi.org/10.1007/s11419-017-0359-8