本研究では市販脱法ハーブ製品に混入される薬物、特に合成カンナビノイドに着目 し、その単離、構造解析、分析方法及び市場調査を通じ、未規制薬物に関する理化学 データを収集した。
市販製品の分析はメタノール抽出液をLC/PDA/MS、GC/MSにより分析し、その際 に未だ国内にて規制がなされていない化合物を発見した際には、各種クロマトグラフ 法により単離精製を実施し、HRMSやNMRにより化学構造を決定した。
17-19 のように塩素原子の由来する位置異性体の混入が懸念される場合には、特に
フッ素化された逆相系カラムによる LC分析により判別する手法を開発した。
また 4及び7のように文献にそのカンナビノイド作用が未記載な化合物が発見・同 定された場合は、G-タンパク結合実験によりカンナビノイド作用の有無を調べ、その 危険性を明らかにした。
合成カンナビノイドの中には抗炎症作用を有するものが報告されていたので、単離 した 合成カンナビノイドのうち被検物質の量が確保できたインドール骨格を有する 23種類の化合物(図.4-2-1-1) について、in vivo における作用を検証する目的で、マウ ス耳殻 TPA法抗炎症活性試験を実施し、抗炎症作用に関する構造活性相関を検討した。
抗炎症試験の結果、naphthoylindoleに活性が多く確認され、そのうち特に JWH-018(1),
-122(5), -210(6) が強い活性を示した。また naphthoylindoleのうち、インドール骨格の
窒素原子からの側鎖及びナフチル基の状態による構造活性相関が確認された。一方、
adamantyl-indole、adamantyl-indazole、benzoylindole、phenylacetylindole及び naphtopyrole 類には殆ど活性が見られなかった。
抗炎症試験にて強い活性が認められた1, 5及び6の3種類の化合物について、マウ ス背部皮膚を用いたDMBA及びTPAによる二段階発癌抑制実験を行ったところ、い ずれも強い抗発癌作用が確認された。これらの知見はインドール類縁化合物の生体に 対する有益な作用であると考えられ、特に癌に対する新たな治療手段の検討・開発に 向け役立つ可能性がある知見が得られた。
［参考文献］
(1) Masako Aragane、Yohei Sasaki、Jun’ichi Nakajima、Nobutaka Fukumori, Masao Yo shizawa、Yukiko Suzuki、Shigemi Kitagawa、Ken’ichiro Mori、 Shuzo Ogino、Ichiro Yasuda and Seiji Nagumo. (2011). Peyote identification on the basis of differences in morphology, mescaline content, and trnL/trnF sequence between Lophophora williamsi i and L. diffusa. J Nat Med., 65:103-110.
(2) 浜野朋子、塩田寛子、中嶋順一、安田一郎. (2007). 市販サルビア葉及び関連製品 に含有される Salvinorin A. 生薬学雑誌61(1):1-5.
(3)荒金眞佐子、中嶋順一、福田達男、吉澤政夫、鈴木幸子、北川重美、安田一郎. (2 008).乱用薬物ハルマラの鑑定と Peganum harmala 中のharmaline 及びharmine 含量.
生薬学雑誌62(2):35-42.
(4) 中嶋順一、荒金眞佐子、浜野朋子、塩田寛子、吉澤政夫、鈴木幸子、北川重美，
安田一郎，森謙一郎，荻野周三(2008). サボテンを用いた植物系ドラッグに含まれる メスカリンの分析. Ann. Rep. Tokyo Metr. Inst. Pub. Health, 59, 91-95.
(5) 吉澤政夫、荒金眞佐子、鈴木幸子、北川重美、中嶋順一、森謙一郎、 荻野周三.
(2011).大麻種子の新たな発芽能力迅速確認法について. 生薬学雑誌65(1):1-9.
(6) 鈴木仁. (2013). 違法ドラッグ分析の現状と課題.ぶんせき①:13-17, 日本分析化学 会.
(7)http://www/yomiuri.co.jp/national/news/20120619-OYT1T00459.htm. Accessed January 2013.
(8)http://sankei.jp.msn.com/affairs/news/120713/crm12071310230009-nl.htm. Accessed January 2013.
(9)http://www/yomiuri.co.jp/entertainment/news2/20121101-OYT8T00929.htm. Accessed J anuary 2013.
(10)http://www/yomdr.yomiuri.co.jp/page.jsp?id=69654. Accessed January 2013.
(11)http://chubu.yomiuri.co.jp/tokushu/watch/watxh121121_1.htm.Accessed January 2013.
(12)http://sankei.jp.msn.com/affairs/news/12110/crm12111018010009-n2.htm. Accessed Jan uary 2013.
(13) 高橋美佐子、中嶋順一、瀬戸隆子、金井千恵子 (2011). 薬物乱用防止 最近の違 法ドラッグ製品に含有される薬物について. 都薬雑誌 33(6):8-12
(14) Ulrich S. Zimmermann, Patricia R. Winkelmann, Max Pilhatsch, Josef A. Nees, R ainer Spanagel, Katja Schulz (2009). Withdrawal Phenomena and Dependence Syndro me After the Consumption of "Spice Gold", Dtsch Arztebl Int., 106(27) 464-467.
(15) EMCDDA-Europol 2010 Annual Report on the implementation of Council
Decision 2005/387/JHA, In accordance with Article 10 of Council Decision 2005/387/J HA on the information exchange, risk-assessment and control of new psychoactive sub stances, European Monitoring Centre for Drugs and Drug Addiction (2010).
(16) Uchiyama N, Kikura-Hanajiri R, Kawahara N, Haishima Y, Goda Y (2009).
Identification of a cannabinoid analog as a new type of designer drug in a herbal pro duct. Chem Pharm Bull., 57: 439–441.
(17) Uchiyama N, Kikura-Hanajiri R, Kawahara N, Goda Y (2009). Identification of a cannabimimetic indole as a designer drug in a herbal product. Forensic Toxicol., 27:
61–66.
(18) Uchiyama N, Kawamura M, Kikura-Hanajiri R, Goda Y (2011). Identification and quantitation of two cannabimimetic phenylacetylindoles JWH-251 and JWH-250, and four cannabimimetic naphthoylindoles JWH-081, JWH-015, JWH-200, and JWH-073 as designer drugs in illegal products. Forensic Toxicol., 29:25-37.
(19) Nakajima J, Takahashi M, Seto T, Suzuki J (2011). Identification and quantitation of cannabimimetic compound JWH-250 as an adulterant in products obtained via the Internet. Forensic Toxicol., 29:51-55.
(20) Nakajima J, Takahashi M, Seto T, Kanai C, Suzuki J, Yoshida M, Hamano T (2011). Identification and quantitation of two benzoylindoles AM-694 and (4-methoxy-phenyl)(1-pentyl-1H-indol-3-yl)methanone, and three cannabimimetic naphthoylindoles JWH-210, JWH-122, and JWH-019 as adulterants in illegal products obtained via the Internet. Forensic Toxicol., 29:95-110.
(21) Nakajima J, Takahashi M, Nonaka R, Seto T, Suzuki J, Yoshida M, Kanai C, Hamano T (2011). Identification and quantitation of a benzoylindole (2-methoxyphenyl) (1-pentyl-1H-indol-3-yl)methanone and a naphthoylindole [1-(5-fluoropentyl-1H-indol-3-yl](naphthalene-1-yl)methanone (AM-2201) found in illegal products obtained via the Internet and their cannabimimetic effects evaluated by in vitro [35S]GTPγS binding assays. Forensic Toxicol., 29:132-141.
(22) Nakajima J, Takahashi M, Seto T, Yoshida M, Kanai C, Suzuki J, Hamano T (2012). Identification and quantitation of two new naphthoylindole drugs-of-abuse, [1-(5-hydroxypentyl)-1H-indol-3-yl](naphthalen-1-yl)methanone (AM-2202) and [1-(4-pentenyl)-1H-indol-3-yl](naphthalen-1-yl)methanone, with other synthetic cannabinoids in unregulated “herbal” products circulated in the Tokyo area. Forensic Toxicol., 30:
33-44
(23) Nakajima J, Takahashi M, Seto T, Yoshida M, Kanai C, Suzuki J, Uemura N, Hamano T (2013). Analysis of azepane isomers of AM-2233, AM-1220 and detection
of an inhibitor of fatty acid amide hydrolase [3’-(aminocarbonyl)(1,1’-biphenyl)-3-yl]-cyclohexylcarbamate (URB597) obtained as designer drugs in Tokyo area. Forensic Toxicol., 31:76-85.
(24) Nakajima J, Nakae D, Yasukawa K (2013). Structure-dependent inhibitory effects of synthetic cannabinoids against 12-O-tetradecanoylphorbol-13-acetate-induced inflam-mation and skin tumor promotion in mice. J. Pharm. Pharmcol. 65:8:1223-1230.
(25) Moosmann B, Kneisel S, Girreser U, Brecht V, Westphal F, Auwarter V.(2012).
Separation and structural characterization of the synthetic cannabinoids JWH-412 and 1-[(5-fluoropentyl)-1H-indol-3yl]-(4-methylnaphthalene-1-yl)methanone using GC-MS, NMR analysis and a flash chromatography system. Forensic Sci. Int., Jul 10;220(1-3):
e17-22. doi:10.1016/j.forsciint.2011.
(26) Uchiyama N, Kawamura M, Kikura-Hanajiri R, Goda Y (2012). Identification of two new-type synthetic cannabinoids, N-(1-adamantyl)-1-pentyl-1H-indole-3-carboxamide (APICA) and N-(1-adamantyl)-1-pentyl-1H-indazole-3-carboxamide (APINACA), and detection of five synthetic cannabinoids, AM-1220, AM-2233. AM-1241, CB-13 (CRA-13), and AM-1248, as designer drugs in illegal products. Forensic Toxicol., 30:114-125.
(27) Kneisel S, Bisel P, Brecht V, Broecker S, Müller M, Auwarter V (2012). Identi-fication of the cannabimimetic AM-1220 and its azepane isomer (N-methylazepan-3-yl) -3-(1-naphthoyl)indole in a research chemical and several herbal mixtures. Forensic Toxicol., 30:126-134.
(28) Huffman J, Szklennik P, Almond A, Bushell K, Selley D, He H, Cassidy M, Wiley J, Martin B. (2005). 1-pentyl-3-phenylacetylindoles, a new class of cannabimime tic indoles. Bioorg Med Chem Lett., 15:4110–4113.
(29) Makriyannis A, Deng H. (2007). Cannabimimetic indole derivatives. United States Patent, US 7,241,799 B2.
(30) Huffman JW, Zengin G, Wu MJ, Lu J, Hynd G, Bushell K, Thompson ALS, Bus hell S, Tartal C, Hurst DP, Reggio PH, Selley DE, Cassidy MP, Wiley JL, Martin BR.
(2005). Structure-activity relationships for 1-alkyl-3-(1-naphthoyl)indoles at the canna-binoid CB1 and CB2 receptors: steric and electronic effects of naphthoyl substituents.
New highly selective CB2 receptor agonists. Bioorg. Med. Chem., 13:89-112.
(31) Makriyannis A, Deng H. (2008). Cannabimimetic indole derivatives, United States Patent, US 0090871 A1.
(32) Wiley JL, Compton DR, Dai D, Lainton JA, Phillips M, Huffman JW, Martin B R. (1998). Structure-activity relationships of indole and pyrrole-derived cannabinoids.
J Pharmacol Exp Ther., 285:995-1004.
(33) Bononi M, Belgi P, Tateo F. (2011). Analytical data for identification of the cannabimimetic phenylacetylindole JWH-203. J Anal toxicol., 35:360-363.
(34) Simon Hudson, John Ramsey, Les King, Sarah Timbers, Steve Maynard, Paul I.
Dargan, David M. Wood (2010). Use of High-Resolution Accurate Mass Spectrometry to Detect Reported and Previousiy Unreported Cannabinomimetics in "Herbal High"
Products, J Analytical Toxicol., 34:252-260.
(35) Monde T, Kamisuki T, Kuroda T, Mikumo K, Ohkawa T, Fukube H.(1996) High-performance liquid chromatographic separation of phenols on a fluorocarbon-bonded sil ica gel column. J Chromatogr A 722:273-280.
(36) Piomelli D, Tarzia G, Duranti A, Tontini A, Mor M, Compton TR, Dasse O, Mo naghan EP, Parrott JA, Putman D. (2006). Pharmacological profile of the selective FAAH inhibitor KDS-4103 (URB597). CNS Drug Rev., 12(1):21-38.
(37) Clapper Jason JR, Mangieri RA, Piomelli D. (2009). The endocannabinoid system as a target for the treatment of cannabis dependence. Neuropharmacology 56:235-243.
(38) Manera C, anera, Tuccinardi T, Martinelli A. (2008). Indole and related compoun ds as cannabinoid Ligands. Med. Chem. 8:370–387.
(39) Willis PG, Katoch-Rouse R, Horti AG. (2003). Regioselective F-18 radiolabeling of AM694, a CB1 cannabinoid receptor ligand. J Label compd Radiopharm., 46:799-804.
(40) Aung MM, Griffin G, Huffman JW, Wu MJ, Keel C, Yang B, Showalter VM, Abood ME, Martin BR. (2000). Influence of the N-1 alkyl chain length of cannabimi metic indoles upon CB1 and CB2 receptor binding. Drug Alcohol Depend., 60:133-140.
(41) Brents Lisa K., Reichard Emily E., Zimmerman Sarah M., Moran Jeffery H., Fan tegrossi William E., Prather Paul L. (2011). Phase I Hydroxylated Metabolites of the K2 Synthetic Cannabinoid JWH-018 Retain In Vitro and In Vivo Cannabinoid 1 Rece-ptor Affinity and Activity PLoS ONE Vol.6 lssue7, e21917. doi:10.1371/journal.pone.0 021917.
(42) Makriyannis A, Deng H. Cannabimimetic indole derivatives. WO 01/28557 A1 (2 001), 1-25, PCT/US00/28832.
(43) Deng H, Gifford, A.M., Zvonok, A.M., et al. (2005). Potent cannabinergic indole analogues as radioiodinatable brain imaging agents for the CB1 cannabinoid receptor.
J. med. Chem. 48:6386-6392.
(44) Matsuda LA, Lolait SJ, Brownstein MJ, Young AC, Bonner TI. (1990). Structure of a cannabinoid receptor and functional expression of the cloned cDNA. Nature; 346:
561-4.
(45) Munro S, Thomas KL, Abu-Shaar M.(1993). Molecular characterization of a peri-pheral receptor for cannabinoids. Nature; 365:61-5.
(46) Fleckenstein, A.E., Metzger, R.R., Wilkins, D.G., et al. (1997). Rapid and Rever-sible Effects of Methamphetamine on Dopamine Transporters. J Pharmacol. Exp. Ther., 282:834-838.
(47) Nagai, F., Nonaka, R., Satoh, K., et al. (2007). The effects of non-medically used psychoactive drugs on monoamine neurotransmission in rat brain. Eur. J. Pharma col., 559:132-137.
(48) Nonaka, R., Nagai, F., Satoh, K., et al. (2007). In vitro screening of psychoact-ive drugs by [35S]GTPγS binding in rat brain membranes. Biol. Pharm. Bull., 30:2328-2333.
(49) 佐藤かな子、野中良一 (2008). 薬物乱用防止を目的とした向精神薬の In vitro ス
クリーニング法. 薬誌, 128, 1771-1782.
(50) Seema S., Jayesh M., Raman D., et al.(1999) Antitumor Activity of Thalidomide in Refractory Multiple Myeloma. N Engl J Med., 341:1565-1571.
(51) Berdyshev E, Boichot E, Corbel M, Germain N, Lagente V. (1998). Effects of cannabinoid receptor ligands on LPS-induced pulmonary inflammation in mice.
Life Sci.63(8):PL125-PL129.
(52) Hanus L, Breuer A, Tchilibon S, Shiloah S, Goldenberg D, Horowitz M, et al.
(1999). HU-308: a specific agonist for CB(2), a peripheral cannabinoid receptor.
Proc Natl Acad Sci USA. 96:14228-14233.
(53) Smith SR, Denhardt G, Terminelli C.(2001). The anti-inflammatory activities of c annabinoid receptor ligands in mouse peritoninitis models. Eur J Pharmacol. 432(1):
107-119.
(54) Massa F, Marsicano G, Hermann H, Cannich A, Monory K, Cravatt BF, et al. (2 004). The endogenous cannabinoid system protects against chronic inflammation.
J Clin Invest. 113:1202-1209.
(55) H. Fujiki, M. Mori, M.Nakayama, M.Tarada and T.Sugimura. (1979). A possible naturally occurring tumor promoter, teleocidin B from Streptomyces. Biochem, Biophys.
Res. Commum., 90;976-983.
(56) K. Yasukawa, M. Takido, M. Takeuchi, K. Nitta. (1988). Effect of flavonoids on tumor promoter̓s activity. Prog Clin Biol Res. 280:247-50.
(57) Toshihiro Akihisa, Ken Yasukawa. (2001). Antitumor-promoting and anti-inflam-matory activities of triterpenoids and sterols from plants and fungi. Atta-ur-Rahman (Ed.) Studies in Natural Products Chemistry. Vol. 25, 43-126.
(58) Ken Yasukawa, Michio Takido, Tetsuro Ikekawa, Fumitake Shimada, Mieko Take-uchi and Shigeki Nakagawa. (1991). Relative Inhibitory Activity of Berberine-Type Alkaloids against 12-O-Tetradecanoylphorbol-13-acetate-Induced Inflammation in Mice.
Chem. Pharm. Bull., 39(6);1462-1465.
(59) K. Yasukawa, T. Akihisa, Y. Kasahara, M. Ukiya, K. kumaki, T. Tamura, S. Ya-manouchi, M. Takido.(1998). Inhibitory effect of heliantriol C; a component of edible Chrysanthemum, on tumor promotion by 12-O-tetradecanoylphorbol-13-acetate in two-stage carcinogenesis in mouse skin. Phytomedicine, 5(3); 215-218.
(60) Hecker E.(1967). Phorbol esters from croton oil chemical nature and biological activities. Naturwissenschaften 54, Issue 11:282-284.
(61) Van Duuren, B.L. (1969). Tumor-promoting agents in two-stage carcinogenesis.
Prog. Exp. Tumor Res. 11:31-68.
(62) Yasukawa K et al.(2009). Relative inhibitory activity of bile aids against 12-O-tetradecanoylphorbol-13-acetate-induced inflammation, and chenodeoxycholic acid inhi-bitttion of tumor promotion in mouse skin two-stage carcinogenesis.
J Pharm Pharmacol. 61:1051-1056.
(63) Junichi Nakajima, Dai Nakae, Ken Yasukawa (2013). Structure-dependent inhibito-ry effects of synthetic cannabinoids against 12-O-tetradecanoylphorbol-13-acetate-induced inflammation and skin tumor promotion in mice.
Journal of Pharmacy and Pharmacolgy, 65(8);1223-1230.
(64) Preet A, Qamri Z, Nasser MW, Prasad A, Shilo K, Zou X, Groopman JE, Ganju RK. (2011). Cannabinoid receptors, CB1 and CB2, as novel targets for inhibitio n of non-small cell lung cancer growth and metastasis. Cancer Prev Res (Phila). Jan;
4(1):65-75.
(65) Qamri Z, Preet A, Naser MW, Bass CE, Leone G, Barsky SH, Ganju RK. (2009).
Synthetic cannabinoid receptor agonists inhibit tumor growth and metastasis of breast cancer. Mol Cancer Ther. Nov;8(11):3117-29.
本要約は以下の論文をまとめたものである。
1. Nakajima J, Takahashi M, Seto T, Suzuki J (2011) Identification and quantitation of cannabimimetic compound JWH-250 as an adulterant in products obtained via the Internet.
Forensic Toxicol.29:51-55.
2. Nakajima J, Takahashi M, Seto T, Kanai C, Suzuki J, Yoshida M, Hamano T (2011) Identification and quantitation of two benzoylindoles AM-694 and
(4-methoxyphenyl)(1-pentyl-1H-indol-3-yl)methanone, and three cannabimimetic naphthoylindoles JWH-210, JWH-122, and JWH-019 as adulterants in illegal products obtained via the Internet. Forensic Toxicol. 29:95-110.
3. Nakajima J, Takahashi M, Nonaka R, Seto T, Suzuki J, Yoshida M, Kanai C, Hamano T (2011) Identification and quantitation of a benzoylindole
(2-methoxyphenyl)(1-pentyl-1H-indol-3-yl)methanone and a naphthoylindole
1-(5-fluoropentyl-1H-indol-3-yl)-(naphthalene-1-yl)methanone (AM-2201) found in illegal products obtained via the Internet and their cannabimimetic effects evaluated by in vitro [35S]GTPγS binding assays. Forensic Toxicol. 29:132-141.
4. Nakajima J, Takahashi M, Seto T, Yoshida M, Kanai C, Suzuki J, Hamano T (2012) Identification and quantitation of two new naphthoylindole drugs-of-abuse,
(1-(5-hydroxypentyl)-1H-indol-3-yl)(naphthalen-1-yl)methanone (AM-2202) and (1-(4-pentenyl)-1H-indol-3-yl)(naphthalen-1-yl)methanone, with other synthetic
cannabinoids in unregulated “herbal” products circulated in the Tokyo area. Forensic Toxicol.
30: 33-44.
5 . Nakajima J, Takahashi M, Seto T, Yoshida M, Kanai C, Suzuki J, Uemura N, Hamano T (2013) Analysis of azepane isomers of AM-2233, AM-1220 and detection of an inhibitor of fatty acid amide hydrolase (3’-(aminocarbonyl)(1,1’-biphenyl)-3-yl)-cyclohexylcarbamate (URB597) obtained as designer drugs in Tokyo area. Forensic Toxicol. 31:76-85.
6. Nakajima J, Nakae D, Yasukawa K (2013) Structure-dependent inhibitory effects of synthetic cannabinoids against 12-O-tetradecanoylphorbol-13-acetate-induced inflammation and skin tumor promotion in mice. J. pharm. Pharmcol. 65:8, 1223-1230.