著 者 は ，LXR 選 択 的 ア ゴ ニ ス ト の 活 性 化 作 用 の 鍵 と な る 光 学 活 性 な ヒ ダ ン ト イ ン (+)-11 の 絶 対 立 体 配 置 を 化 合 物 (+)-11·HBr の Ｘ 線 結 晶 構 造 解 析 の 結 果 か ら S と 決 定 し た ． ま た ，D-(−)-マ ン デ ル 酸 を 用 い た ア ミ ノ 酸 メ チ ル エ ス テ ル 13 の 光 学 分 割 を 経 由 し た 光 学 活 性 な ヒ ダ ン ト イ ン (S)-(+)-11 の 大 量 合 成 法 を 確 立 し た ．さ ら に ，化 合 物 (S)-(−)-10 の 合 成 法 を 改 善 し ，数 百 グ ラ ム ス ケ ー ル の (S)-(−)-10 を 製 造 可 能 な 効 率 的 な 合 成 法 を 開 発 し た ．
105
結 語
核 内 受 容 体 LXR 選 択 的 ア ゴ ニ ス ト に よ る LXR 活 性 化 作 用 に 伴 い ， 血 中 ABCA1 mRNA の 発 現 を 上 昇 さ せ ，HDL-C を 増 加 さ せ ，RCT を 促 進 す る こ と に よ っ て 脂 質 沈 着 抑 制 作 用 を 示 す 新 規 動 脈 硬 化 治 療 薬 の 創 出 を 目 的 に 研 究 を お こ な っ た ．
第 一 章 で は ，ハ イ ス ル ー プ ッ ト ス ク リ ー ニ ン グ (HTS) に よ っ て ，得 ら れ た 化 合 物 1 か ら リ ー ド 化 合 物 4 を 見 出 し た 経 緯 ，そ の 構 造 活 性 相 関 お よ び in vivo 薬 理 評 価 に つ い て 述 べ た ．HTS に よ り 得 た ヒ ッ ト 化 合 物 1 と 他 社 化 合 物 情 報 を 活 用 し た 独 自 の
‘head-to-tail’の ド ラ ッ グ デ ザ イ ン に よ り ，head 部 分 に 2-オ キ ソ ク ロ メ ン 構 造 を ，tail 部 分 に ヒ ダ ン ト イ ン 構 造 を も つ LXR 選 択 的 ア ゴ ニ ス ト 2 を 見 出 し た ．化 合 物 2 は ， 動 脈 硬 化 モ デ ル で の in vivo 薬 理 評 価 に お い て ，300 mg/kg の 経 口 投 与 に て 脂 質 沈 着 抑 制 作 用 を 示 し ，HDL-C の 上 昇 作 用 が 確 認 さ れ て い る こ と か ら 末 梢 血 中 で の コ レ ス テ ロ ー ル 逆 転 送 系 に よ る 直 接 的 な 作 用 と 推 察 さ れ た ． 一 方 で ， 代 謝 安 定 性 お よ び 血 中 濃 度 推 移 に 改 善 の 必 要 性 が あ っ た た め ， こ れ ら の 課 題 の 改 善 を 目 指 し ，head 部 分 を 2-オ キ ソ ク ロ メ ン 構 造 (2) か ら 1,3-ジ ヒ ド ロ イ ソ ベ ン ゾ フ ラ ン 構 造 (3)， さ ら に ， 1,1-ビ ス(ト リ フ ル オ ロ メ チ ル)カ ル ビ ノ ー ル 構 造 (4) へ と 換 え た と こ ろ ， 代 謝 安 定 性 お よ び 血 中 濃 度 の 改 善 だ け で な く ，LXR 活 性 化 作 用 の 向 上 に 成 功 し た ．こ こ で ，LXR 活 性 化 作 用 の 鍵 と な る ‘His435-Trp457 activation switch’ の 相 互 作 用 が ド ッ キ ン グ モ デ ル か ら 推 察 さ れ た ．LXR 選 択 的 ア ゴ ニ ス ト 4 は ，動 脈 硬 化 モ デ ル で の in vivo 薬 理 評 価 に お い て ，100 mg/kg 経 口 投 与 に て 脂 質 沈 着 抑 制 作 用 を 示 し ，か つ ，LXR ア ゴ ニ
Figure 57. Summary of structural modification in hit to lead.
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ス ト の 副 作 用 と し て 懸 念 さ れ る TG の 増 加 を 抑 制 で き る こ と が 判 明 し ，リ ー ド 化 合 物 と し て 位 置 付 け た (Figure 57)．
第 二 章 で は ， リ ー ド 化 合 物 4 の 各 鏡 像 異 性 体 の 活 性 化 作 用 の 確 認 ， 絶 対 立 体 配 置 の 決 定 お よ び 光 学 活 性 な ヒ ダ ン ト イ ン (+)-5 の 製 造 法 を 確 立 し た 経 緯 に つ い て 述 べ た ． さ ら に ， 化 合 物 4 の 代 謝 物 の 同 定 に つ い て 述 べ た ． 第 一 に ， ヒ ダ ン ト イ ン 5 位 に 不 斉 炭 素 を 有 す る こ と か ら 各 鏡 像 異 性 体 の 活 性 化 作 用 を 評 価 し ，(+)-4 に 所 望 の 活 性 化 作 用 を 有 す る こ と を 明 ら か に し た ．化 合 物 4 の 絶 対 立 体 配 置 に つ い て は ，ヒ ダ ン ト イ ン 体 (+)-Br-6 お よ び (+)-Cl-6 の X 線 結 晶 構 造 解 析 の 結 果 を も と に ，S と 決 定 し た ． 光 学 活 性 な ヒ ダ ン ト イ ン (+)-5 は ， 中 間 体 で あ る ア ミ ノ 酸 エ ス テ ル に 対 し て L-(+)-マ ン デ ル 酸 を 用 い た 光 学 分 割 を 経 由 し ，安 定 に 大 量 供 給 で き る 合 成 法 を 確 立 し た ． 第 二 に ， 化 合 物 4 の 血 中 濃 度 推 移 を 確 認 し た と こ ろ ， 化 合 物 2 と 比 べ て 改 善 は し た も の の リ ン カ ー 部 位 で 酸 化 的 代 謝 を 受 け ，カ ル ボ ン 酸 9 を 生 成 す る こ と を 明 ら か に し た ． 化 合 物 4 の 血 中 濃 度 は ， 化 合 物 2 と 比 べ て 改 善 さ れ て は い る も の の 決 し て 十 分 で は な い こ と の 原 因 が ， こ の 代 謝 に あ る こ と が 明 ら か に な っ た ．
第 三 章 で は ，‘head-to-tail’の ド ラ ッ グ デ ザ イ ン を 基 に ，カ ル ビ ノ ー ル 構 造 (head 部 分) と ヒ ダ ン ト イ ン 構 造 (tail 部 分) を 結 ぶ リ ン カ ー 部 分 に 有 益 な 2-ヒ ド ロ キ シ ア セ ト フ ェ ノ ン 構 造 を 有 す る 化 合 物 10 を 見 出 し た 経 緯 ， お よ び そ の 構 造 活 性 相 関 に つ い て 述 べ た ．す な わ ち ，構 造 活 性 相 関 の 結 果 ，化 合 物 4 と 比 べ て 化 合 物 (−)-10 は ，LXR
活 性 化 作 用 (EC5 0) お よ び 選 択 性 ( selectivity for EC5 0) の さ ら な る 改 善 だ け で な く ，代 謝 安 定 性 お よ び 血 中 濃 度 推 移 も 改 善 さ れ る こ と が 明 ら か に な っ た ．化 合 物 10 の 鏡 像 異 性 体 の う ち 良 好 な (−)-10 は ， 動 脈 硬 化 モ デ ル と し て 知 ら れ る LDL 受 容 体 欠 損 マ ウ ス で の 薬 効 試 験 の 結 果 に お い て ， 脂 質 沈 着 抑 制 作 用 を 示 し た ． 血 中 ABCA1 mRNA の 発 現 が 上 昇 し ，HDL-C が 増 加 し て い る こ と か ら ，RCT の 亢 進 に よ る 抗 動 脈 硬 化 作 用 に 繋 が る 可 能 性 が 示 唆 さ れ た ． こ れ ら の 結 果 か ら ， 化 合 物 (−)-10 は 新 規 な
Figure 58. Summary of structural modification in lead optimization.
107
動 脈 硬 化 治 療 薬 と し て 期 待 さ れ ， さ ら な る 薬 理 ， 薬 物 動 態 お よ び 安 全 性 評 価 の 候 補 化 合 物 と し て 位 置 付 け た (Figure 58)．
第 四 章 で は ， 候 補 化 合 物 (−)-10 の 絶 対 立 体 配 置 の 決 定 ，(−)-10 を 合 成 す る た め の 中 間 体 と な る 光 学 活 性 な ヒ ダ ン ト イ ン (S)-(+)-11 の 大 量 合 成 法 の 確 立 お よ び 化 合 物 (S)-(−)-10 の 効 率 的 な 合 成 法 の 確 立 に つ い て 述 べ た ．LXR 選 択 的 ア ゴ ニ ス ト 活 性 化 作 用 の 鍵 と な る 合 成 中 間 体 で あ る 光 学 活 性 な ヒ ダ ン ト イ ン (+)-11 の 絶 対 立 体 配 置 は ， 化 合 物 (+)-11·HBr を 用 い て ，X 線 結 晶 構 造 解 析 を し ，S と 決 定 し た ． ま た ，D-(−)-マ ン デ ル 酸 を 用 い た ア ミ ノ 酸 エ ス テ ル 13 の 光 学 分 割 を 経 由 し た 光 学 活 性 な ヒ ダ ン ト イ ン (S)-(+)-11 の 大 量 合 成 法 ， お よ び (S)-(−)-10 の 安 定 供 給 可 能 な 合 成 法 を 確 立 し た ．
す な わ ち ，D-(−)-マ ン デ ル 酸 を 用 い た 光 学 分 割 を 経 て ア ミ ノ 酸 エ ス テ ル (+)-13 を 高 い 鏡 像 体 過 剰 率 で 大 量 に 供 給 す る 方 法 を 確 立 し ，そ れ に よ り (S)-(+)-11 の 大 量 合 成 が 可 能 に な っ た ．さ ら に ，こ の 光 学 活 性 ヒ ダ ン ト イ ン と 合 成 容 易 な ア ニ リ ン 誘 導 体 14 お よ び ア セ ト フ ェ ノ ン 誘 導 体 17 を 利 用 し て ，(S)-(−)-10 を 安 定 に 供 給 で き る 合 成 経 路 を 確 立 し た ．
本 論 文 は ，動 脈 硬 化 治 療 薬 の 開 発 を 目 的 と し た LXR ア ゴ ニ ス ト の 創 薬 研 究 で あ り ， 見 出 さ れ た 高 活 性 か つ 高 い 選 択 性 を 示 す LXR ア ゴ ニ ス ト (S)-(−)-10 は 現 在 に お い て 最 も 有 効 な 化 合 物 の 一 つ で あ る ． ま た ，in vitro レ ポ ー タ ー 遺 伝 子 ア ッ セ イ で の LXR 選 択 性 が ，in vivo 薬 理 評 価 に お い て TG の 増 加 の 副 作 用 を 抑 制 で き る こ と を 証 明 し た 初 の 報 告 例 で あ る ．候 補 化 合 物 (S)-(−)-10 に 至 る ま で の 構 造 活 性 相 関 は ，今 後 の LXR 選 択 的 ア ゴ ニ ス ト の 創 薬 研 究 に 有 益 な 情 報 を 寄 与 す る も の と 考 え る ．ま た ， 化 合 物 (S)-(−)-10 の 改 良 製 造 法 は ，今 後 の さ ら な る 薬 理 ，薬 物 動 態 や 安 全 性 評 価 の た め の 原 薬 の 安 定 供 給 を 可 能 と し ， さ ら な る 化 合 物 解 析 に 寄 与 す る も の と 考 え る ． さ ら に ， 光 学 活 性 な ヒ ダ ン ト イ ン の 絶 対 立 体 配 置 の 決 定 ， お よ び ア ミ ノ 酸 エ ス テ ル の マ ン デ ル 酸 を 用 い た 光 学 分 割 法 は ， ア ミ ノ 酸 誘 導 体 の 構 造 決 定 や 大 量 合 成 に お い て も 有 用 で あ る ．
な お ，化 合 物 (S)-(−)-10 は ，安 全 性 評 価 に お い て 予 期 せ ぬ 副 作 用 が 確 認 さ れ た た め ， そ の 後 の 開 発 を 断 念 し た (詳 細 は 契 約 お よ び 権 利 上 不 記 載)．
108
実験の部
General procedure
Commercially available reagents and solvents were used without further purification. Thin layer chromatography (TLC) analyses were performed on silica gel 60 F254 plates (Merck). 1H NMR and 13C NMR spectra were obtained on a JEOL JNM-LA or ECS 400 MHz spectrometer using CDCl3, CD3OD or d6-DMSO as the solvent with tetramethylsilane as the internal standard. Infrared (IR) spectra were recorded on a Thermo Nicolet 370 FT-IR (KBr or ATR) spectrometer. Mass spectra were obtained on a JEOL MS-BU20 mass spectrometer.
Elemental analyses (C, H, N) were performed using a Yanaco MT-5 instrument. Melting points were determined in open glass capillaries on a Buchi B-545 melting point apparatus. Optical rotations were measured on a JASCO P2200 polarimeter operating at the sodium D line at room temperature. The chiral HPLC analyses were performed on a Shimazu LC-2010A HT liquid chromatograph.
Chapter-1
Section-1
Chemical Experimental Details
Scheme 7
1,3-Bis(allyloxy)benzene (31) 77):
To a stirred suspension of 1,3-dihydroxybenzene (30) (5.0 g, 45 mmol) and K2CO3 (19 g, 136 mmol) in DMF (50 mL), allyl chloride (12 g, 154 mmol) was slowly added at room temperature. The reaction mixture was stirred at 70 °C for 24 h, and then diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4, and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/ EtOAc = 2/1) to give the title compound (7.3 g, 85%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 4.59 (4H, d, J = 7.4 Hz), 5.36 (2H, dd, J = 2.9, 11.9 Hz), 5.49 (2H, dd, J = 2.9, 17.3 Hz), 6.08–6.18 (2H, m), 6.59 (1H, s), 6.60 (2H, d, J = 7.1 Hz), 7.24 (1H, t, J = 7.1 Hz); MS (EI) m/z 190 [M]+.
2,4-Diallylbenzene-1,3-diol (32) 77):
A stirred solution of 31 (7.3 g, 39 mmol) in N,N-dimethylaniline (60 mL) was heated at 200 °C for 16 h. The reaction mixture was allowed to cool to room temperature. To this mixture, hexane (200 mL) and 2 N NaOH aq.
109
(150 mL) was diluted. The organic layer was separated and washed with 2 N NaOH aq.. The aqueous layer was neutralized with 2 N HCl until pH 7 and extracted with Et2O. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc
= 12/1) to give the title compound (4.2 g, 57%) as a yellow oil; 1H NMR (400 MHz, CDCl3) δ 3.34 (2H, d, J = 6.4 Hz), 3.48 (2H, d, J = 6.4 Hz), 5.06 (1H, s), 5.11–5.26 (4H, m), 5.26 (1H, s), 5.95–6.05 (2H, m), 6.39 (1H, d, J = 8.3 Hz), 6.85 (1H, d, J = 8.3 Hz); MS (EI) m/z 190 [M]+.
2,4-Dipropylbenzene-1,3-diol (33) 77):
A suspension of 32 (4.2 g, 22 mmol) and 10% Pd/C (420 mg) in MeOH (30 mL) was stirred under a hydrogen atmosphere at room temperature for 18 h. The reaction mixture was filtered through a pad of Celite. The filtrate was concentrated in vacuo to give the title compound (4.2 g, 98%) as pale yellow powders; 1H NMR (400 MHz, CDCl3) δ 0.97 (3H, t, J = 7.3 Hz), 0.99 (3H, t, J = 7.3 Hz), 1.55–1.65 (4H, m), 2.49 (2H, t, J = 7.6 Hz), 2.61 (2H, t, J = 7.6 Hz), 4.55 (1H, s), 4.69 (1H, s), 6.33 (1H, d, J = 8.0 Hz), 6.81 (1H, d, J = 8.0 Hz); MS (EI) m/z 194 [M]+.
7-Hydroxy-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-2-one (34):
A stirred mixture of 33 (4.2 g, 22 mmol), zinc chloride (3.5 g, 26 mmol) and ethyl 4,4,4-trifluoro-3-oxobutanoate (12 g, 65 mmol) was heated at 110 C for 18 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4, and concentrated in vacuo. The residue was solidified from acetone and hexane to give the title compound (4.0 g, 59%) as pale yellow powders; 1H NMR (400 MHz, CDCl3) δ 1.00 (3H, t, J = 7.2 Hz), 1.01 (3H, t, J = 7.2 Hz), 1.61–1.70 (4H, m), 2.64 (2H, t, J = 7.6 Hz), 2.83 (2H, t, J = 7.6 Hz), 5.36 (1H, s), 6.61 (1H, s), 7.33 (1H, s); MS (EI) m/z 314 [M]+.
Scheme 8
1,3-Dimethoxy-2-propylbenzene (36) 78):
To a solution of 1,3-dimethoxybenzene (35) (10 g, 72 mmol) in THF (100 mL), a solution of n-BuLi in hexane (50 mL, 1.6 M solution) was added dropwise at 0 °C. After stirring at 0 °C for 2 h, to this solution was added a solution of 1-iodopropane (12 g, 73 mmol) in THF (20 mL). The reaction mixture was allowed to warm to room temperature for 22 h and then water was added to this mixture. The organic layer was separated and the aqueous
110
layer was extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc = 60/1) to give the title compound (5.9 g, 45%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.2 Hz), 1.50 (2H, qt, J = 7.2, 7.6 Hz), 2.62 (2H, t, J = 7.6 Hz), 3.80 (6H, s), 6.52 (2H, d, J = 8.0 Hz), 7.10 (1H, t, J = 8.0 Hz); MS (EI) m/z 180 [M]+.
2-Propylbenzene-1,3-diol (37) 78):
To a solution of 36 (5.9 g, 33 mmol) in CH2Cl2 (50 mL), a solution of boron tribromide in CH2Cl2 (90 mL, 1.0 M solution) was added over a period of 45 min at −70 °C. After stirring at −70 °C for 1 h, the reaction mixture was allowed to warm to room temperature for 2 h and then poured into ice water. The organic layer was separated and the aqueous layer was extracted with CHCl3. The combined organic layers were washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc
= 8/1) to give the title compound (3.8 g, 76%) as colorless powders; 1H NMR (400 MHz, CDCl3) δ 0.99 (3H, t, J
= 7.2 Hz), 1.60 (2H, qt, J = 7.2, 7.6 Hz), 2.61 (2H, t, J = 7.6 Hz), 4.68 (2H, brs), 6.38 (2H, d, J = 8.0 Hz), 6.92 (1H, t, J = 8.0 Hz); MS (EI) m/z 152 [M]+.
7-Hydroxy-8-propyl-4-(trifluoromethyl)-2H-chromen-2-one (38):
A stirred mixture of 37 (3.7 g, 24 mmol), zinc chloride (3.6 g, 26 mmol) and ethyl 4,4,4-trifluoro-3-oxobutanoate (4.9 g, 26 mmol) was heated at 110 C for 18 h and then diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (CHCl3). Thus obtained product was crystallized from CHCl3 to give the title compound (5.0 g, 77%) as pale brown powders; 1H NMR (400 MHz, CDCl3) δ 1.00 (3H, t, J = 7.4 Hz), 1.65 (2H, qt, J = 7.4, 7.8 Hz), 2.84 (2H, t, J = 7.8 Hz), 5.95 (1H, s), 6.63 (1H, s), 6.85 (1H, d, J = 8.6 Hz), 7.48 (1H, dd, J = 8.6 Hz); 13C NMR (100 MHz, CDCl3) δ 14.7, 22.8, 25.4, 108.1, 112.6 (q, J = 5.9 Hz), 113.8, 118.1, 122.6 (q, J = 276 Hz), 124.5, 142.8 (q, J = 32.4 Hz), 154.7, 158.6, 160.6; MS (EI) m/z 272 [M]+.
111 Scheme 10
Synthetic procedure of the hydantoin derivatives
5-(4-Methoxyphenyl)-5-methylimidazolidine-2,4-dione (42a) 79):
To a stirred solution of 1-(4-methoxyphenyl)ethanone (41a) (4.0 g, 33 mmol) in EtOH (6.0 mL), NaCN (2.0 g, 50 mmol), (NH4)2CO3 (9.0 g, 117 mmol), and water (6.0 mL) were added at room temperature. The reaction mixture was irradiated in a microwave (Initiator; Biotage AB) at 100 C for 1 h and then concentrated in vacuo. The residue was filtered off and washed with water. The resultant solid was then crystallized from a mixture solvent (hexane/EtOAc = 4/1) to give the title compound (3.5 g, 47%) as colorless crystals; mp 207.4–211.7 °C; 1H NMR (400 MHz, CD3OD) δ 1.73 (3H, s), 3.79 (3H, s), 6.93 (2H, d, J = 8.8 Hz), 7.41 (2H, d, J = 8.8 Hz);MS (EI) m/z 220 [M]+.
Compounds 42b~k were prepared in the same manner as the synthesis of 42a.
Characterization data
The NMR and MS data of the compounds 42b~k are described below.
5-Methyl-5-(p-tolyl)imidazolidine-2,4-dione (42b):
Compoud 42b was prepared from 1-(p-tolyl)ethan-1-one in a manner similar to that described for compound 42a.
The title compound was obtained as a white solid; 1H NMR (400 MHz, d6-DMSO) δ 1.74 (3H, s), 2.32 (3H, s), 7.20 (2H, d, J = 8.9 Hz), 7.38 (2H, d, J = 8.9 Hz); MS (EI) m/z 204 [M]+.
5-(2-Methoxyphenyl)-5-methylimidazolidine-2,4-dione (42c):
Compound 42c was prepared from 1-(2-methoxyphenyl)ethan-1-one in a manner similar to that described for compound 42a. The title compound was obtained as a white solid; 1H NMR (400 MHz, d6-DMSO) δ 1.63 (3H, s), 3.71 (3H, s), 6.90 (1H, dd, J = 1.6, 7.8 Hz), 6.99 (1H, d, J = 1.6 Hz), 7.04 (1H, d, J = 7.8 Hz), 7.32 (1H, dd, J = 7.8, 7.8 Hz), 8.59 (1H, s), 10.7 (1H, s); MS (EI) m/z 220 [M]+.
112 5-(3-Methoxyphenyl)-5-methylimidazolidine-2,4-dione (42d):
Compound 42d was prepared from 1-(3-methoxyphenyl)ethan-1-one in a manner similar to that described for compound 42a. The title compound was obtained as a pale yellow oil; 1H NMR (400 MHz, d6-DMSO) δ 1.63 (3H, s), 3.76 (3H, s), 6.94–7.04 (2H, m), 7.32–7.40 (2H, m), 7.93 (1H, s), 10.6 (1H, s); MS (EI) m/z 220 [M]+.
5-(3,4-Dimethoxyphenyl)-5-methylimidazolidine-2,4-dione (42e):
Compound 42e was prepared from 1-(3,4-dimethoxyphenyl)ethan-1-one in a manner similar to that described for compound 42a. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, d6-DMSO) δ 1.62 (3H, s), 3.74 (3H, s), 3.76 (3H, s), 6.93–7.01 (3H, m), 8.58 (1H, s), 10.7 (1H, s);MS (EI) m/z 250 [M]+.
5-(Benzo[d][1,3]dioxol-5-yl)-5-methylimidazolidine-2,4-dione (42f):
Compound 42f was prepared from 1-(benzo[d][1,3]dioxol-5-yl)ethan-1-one in a manner similar to that described for compound 42a. The title compound was obtained as a white solid; 1H NMR (400 MHz, d6-DMSO) δ 1.60 (3H, s), 6.02 (2H, s), 6.90 (1H, d, J = 8.4 Hz), 6.94 (1H, dd, J = 1.4, 8.4 Hz), 6.99 (1H, d, J = 1.4 Hz), 8.56 (1H, s), 10.7 (1H, s);MS (EI) m/z 234 [M]+.
5-(4-(Dimethylamino)phenyl)-5-methylimidazolidine-2,4-dione (42g):
Compound 42g was prepared from 1-(4-(dimethylamino)phenyl)ethan-1-one in a manner similar to that described for compound 42a. The title compound was obtained as a white solid; 1H NMR (400 MHz, d6-DMSO) δ 1.59 (3H, s), 2.87 (6H, s), 6.71 (2H, d, J = 8.6 Hz), 7.23 (2H, d, J = 8.6 Hz), 8.45 (1H, s), 10.6 (1H, s);MS (EI) m/z 233 [M]+.
5-Methyl-5-(4-(trifluoromethyl)phenyl)imidazolidine-2,4-dione (42h):
Compound 42h was prepared from 1-(4-(trifluoromethyl)phenyl)ethan-1-one in a manner similar to that described for compound 42a. The title compound was obtained as a white solid; 1H NMR (400 MHz, d6-DMSO) δ 1.70 (3H,
113
s), 7.72 (2H, d, J = 8.6 Hz), 7.79 (2H, d, J = 8.6 Hz), 8.73 (1H, s), 10.7 (1H, s);MS (EI) m/z 258 [M]+.
5-Methyl-5-(4-nitrophenyl)imidazolidine-2,4-dione (42i):
Compound 42i was prepared from 1-(4-nitrophenyl)ethan-1-one in a manner similar to that described for compound 42a. The title compound was obtained as a brown solid; 1H NMR (400 MHz, d6-DMSO) δ 1.71 (3H, s), 7.78 (2H, d, J = 8.6 Hz), 8.27 (2H, d, J = 8.6 Hz), 8.82 (1H, s), 11.0 (1H, s);MS (EI) m/z 235 [M]+.
tert-Butyl 4-(4-methyl-2,5-dioxoimidazolidin-4-yl)benzoate (42j):
Compound 42j was prepared from tert-butyl 4-acetylbenzoate in a manner similar to that described for compound 42a. The title compound was obtained as a white solid; 1H NMR (400 MHz, d6-DMSO) δ 1.54 (9H, s), 1.67 (3H, s), 7.61 (2H, t, J = 8.6 Hz), 7.92 (2H, d, J = 8.6 Hz), 8.72 (1H, s), 10.9 (1H, s);MS (EI) m/z 290 [M]+.
1-(4-((Tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethan-1-one (41k):
A solution of 1-(4-hydroxyphenyl)ethan-1-one (1.0 g, 7.3 mmol), DHP (741 mg, 8.8 mmol) and p-TsOH·H2O (279 mg, 1.5 mmol) was stirred at room temperature for 22 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo.
The residue was purified by silica gel column chromatography (hexane/EtOAc = 5/1) to give the title compound (554 mg, 34%) as a white solid; 1H NMR (400 MHz, CDCl3) δ 1.53–1.75 (3H, m), 1.84–2.05 (3H, m), 2.52 (3H, s), 3.58–3.62 (1H, m), 3.78–3.87 (1H, m), 5.49 (1H, s), 7.06 (2H, t, J = 8.6 Hz), 7.90 (2H, d, J = 8.6 Hz); MS (EI):
m/z 220 [M]+.
5-Methyl-5-(4-((tetrahydro-2H-pyran-2-yl)oxy)phenyl)imidazolidine-2,4-dione (42k):
Compound 42k was prepared from compound 41k in a manner similar to that described for compound 42a. The title compound was obtained as a white solid; 1H NMR (400 MHz, CDCl3) δ 1.48–1.72 (6H, m), 1.84–1.98 (3H, m), 3.54–3.59 (1H, m), 3.80–3.87 (1H, m), 5.40 (1H, s), 7.00 (2H, t, J = 8.6 Hz), 7.36 (2H, d, J = 8.6 Hz), 7.51 (1H, s) , 9.53 (1H, s);MS (EI) m/z 290 [M]+.
114 Scheme 9
7-(4-Bromobutoxy)-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-2-one (39):
To a suspension of 34 (500 mg, 1.6 mmol) and K2CO3 (330 mg, 2.4 mmol) in DMF (5.0 mL), 1,4-dibromobutane (2.8 mL, 13 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 18 h and diluted with water. The reaction mixture was extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc = 10/1) to give the title compound (689 mg, 96%) as a yellow oil; 1H NMR (400 MHz, CDCl3) δ 0.99 (3H, t, J = 7.3 Hz), 1.02 (3H, t, J = 7.3 Hz), 1.62–1.72 (4H, m), 1.96–2.06 (2H, m), 2.12–2.22 (4H, m), 2.64 (2H, t, J = 7.6 Hz), 2.80 (2H, t, J = 7.6 Hz), 3.54 (2H, t, J = 6.5 Hz), 3.87 (2H, t, J = 5.4 Hz), 6.69 (1H, s), 7.38 (1H, s); MS (EI) m/z 448 [M]+.
7-(4-Bromobutoxy)-8-propyl-4-(trifluoromethyl)-2H-chromen-2-one (40):
To a stirred suspension of 38 (16 g, 58 mmol) and K2CO3 (12 g, 88 mmol) in DMF (80 mL), 1,4-dibromobutane (126 g, 584 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 21 h and then diluted with water. The reaction mixture was extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc = 10/1) to give the title compound (19 g, 80%) as a yellow oil; 1H NMR (400 MHz, CDCl3) δ 0.98 (3H, t, J = 7.4 Hz), 1.59 (2H, qt, J = 7.4, 7.6 Hz), 1.98–2.16 (4H, m), 2.83 (2H, t, J = 7.6 Hz), 3.53 (2H, t, J = 6.2 Hz), 4.14 (2H, t, J = 5.7 Hz), 6.59 (1H, s), 6.90 (1H, d, J = 9.5 Hz), 7.53–7.55 (1H, m); 13C NMR (100 MHz, CDCl3) δ 14.1, 22.2, 24.8, 27.8, 29.4, 33.1, 67.7, 107.3, 108.4, 112.2 (q, J = 5.7 Hz), 119.6, 121.8 (q, J = 276 Hz), 123.8, 141.8 (q, J = 32.4 Hz), 153.4, 159.7, 160.4; MS (EI) m/z 406 [M]+.
5-(4-Methoxyphenyl)-5-methyl-3-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (2):
To a stirred suspension of 5-(4-methoxyphenyl)-5-methylimidazolidine-2,4-dione (42a) (15 g, 70 mmol) and K2CO3 (13 g, 93 mmol) in DMF (200 mL), a solution of 40 (19 g, 46 mmol) in DMF (100 mL) was added dropwise at 0 °C. The reaction mixture was stirred at room temperature for 16 h and then diluted with water at
115
0 °C and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc = 5/2) to give the title compound (18 g, 72%) as colorless crystals; mp 93.2–94.3 °C; IR (KBr) 3269, 2957, 2937, 2869, 1718, 1607, 1279, 1120 cm-1; 1H NMR (400 MHz, CDCl3) δ 0.92 (3H, t, J = 7.6 Hz), 1.55 (2H, qt, J = 7.6, 7.6 Hz), 1.81–1.86 (7H, m), 2.80 (2H, t, J = 7.6 Hz), 3.61 (2H, t, J = 6.0 Hz), 3.79 (3H, s), 4.07 (2H, t, J = 5.6 Hz), 6.12 (1H, brs), 6.61 (1H, s), 6.84 (1H, d, J = 8.8 Hz), 6.90 (2H, d, J = 8.8 Hz), 7.40 (2H, d, J = 8.8 Hz), 7.52 (1H, d, J = 8.8 Hz);
13C NMR (100 MHz, CDCl3) δ 14.0, 22.1, 24.6, 25.4, 26.2, 38.1, 55.3, 63.2, 67.7, 107.2, 108.4, 112.0 (q, J = 5.8 Hz), 114.2 (2C), 119.5, 121.7 (q, J = 275 Hz), 123.6, 123.7, 126.4 (2C), 130.6, 141.8 (q, J = 32.5 Hz), 153.3, 156.8, 159.6, 159.7, 160.4, 175.6; MS m/z 546 [M]+; Anal. Calcd for C28H29F3N2O6: C, 61.53; H, 5.35; N, 5.13.
Found: C, 61.36; H, 5.34; N, 5.20.
Synthetic procedure of the 2-oxochromene derivatives having the dipropyl group
Compounds 62, 63, 43~61 were prepared in the same manner as the synthesis of 2.
Characterization data
The NMR and MS data of the compounds 62, 63, 43~61 are described below.
3-(4-(2-Oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)thiazolidine-2,4-dione (43):
Compound 43 was prepared from compound 39 and thiazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.98 (3H, t, J
= 7.6 Hz), 1.00 (3H, t, J = 7.2 Hz), 1.60–1.70 (4H, m), 1.84–1.91 (4H, m), 2.63 (2H, t, J = 7.6 Hz), 2.79 (2H, t, J
= 7.6 Hz), 3.76 (2H, t, J = 6.8 Hz), 3.84 (2H, t, J = 6.0 Hz), 3.98 (2H, s), 6.69 (1H, s), 7.34 (1H, s); MS (EI) m/z 485 [M]+.
1-Methyl-3-(4-((2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yl)oxy)butyl)imidazolidine-2,4- dione (44):
Compound 44 was prepared from compound 39 and 1-methylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ
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0.98 (3H, t, J = 7.6 Hz), 1.00 (3H, t, J = 7.2 Hz), 1.62–1.68 (4H, m), 1.87–1.89 (4H, m), 2.63 (2H, t, J = 7.6 Hz), 2.79 (2H, t, J = 7.6 Hz), 3.07 (3H, s), 3.63 (2H, t, J = 6.6 Hz), 3.84 (2H, t, J = 5.6 Hz), 3.89 (2H, s), 6.69 (1H, s), 7.38 (1H, s); MS (EI) m/z 482 [M]+.
1-(4-(2-Oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)pyrrolidine-2,5-dione (45):
Compound 45 was prepared from compound 39 and pyrrolidine-2,5-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.98 (3H, t, J
= 7.2 Hz), 1.00 (3H, t, J = 7.6 Hz), 1.60–1.70 (4H, m), 1.83–1.87 (4H, m), 2.63 (2H, t, J = 7.6 Hz), 2.73 (4H, s), 2.79 (2H, t, J = 8.0 Hz), 3.63 (2H, t, J = 6.0 Hz), 3.83 (2H, t, J = 5.6 Hz), 6.69 (1H, s), 7.38 (1H, s); MS (EI) m/z 467 [M]+.
3-(4-(2-Oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)oxazolidin-2-one (46):
Compound 46 was prepared from compound 39 and oxazolidin-2-one in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.98 (3H, t, J = 7.3 Hz), 1.01 (3H, t, J = 7.3 Hz), 1.63–1.68 (4H, m), 1.82–1.89 (4H, m), 2.64 (2H, t, J = 7.6 Hz), 2.80 (2H, t, J = 7.8 Hz), 3.61 (2H, t, J = 6.6 Hz), 3.84 (2H, t, J = 5.9 Hz), 4.93–5.01 (4H, m), 6.69 (1H, s), 7.38 (1H, s); MS (EI) m/z 455 [M]+.
1-(4-(2-Oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)pyrrolidin-2-one (47):
Compound 47 was prepared from compound 39 and pyrrolidin-2-one in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.99 (3H, t, J = 7.3 Hz), 1.01 (3H, t, J = 7.3 Hz), 1.63–1.71 (6H, m), 1.80–1.87 (2H, m), 1.92–2.18 (4H, m), 2.65 (2H, t, J = 7.6 Hz), 2.81 (2H, t, J = 7.6 Hz), 3.54 (2H, t, J = 6.4 Hz), 3.78 (2H, t, J = 6.8 Hz), 3.88 (2H, t, J = 6.4 Hz), 6.69 (1H, s), 7.39 (1H, s); MS (EI) m/z 453 [M]+.
117
1,5-Dimethyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)imidazolidine-2,4-dio ne (48):
Compound 48 was prepared from compound 39 and 1,5-dimethylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.98 (3H, t, J = 7.3 Hz), 1.00 (3H, t, J = 7.3 Hz), 1.45 (3H, d, J = 6.8 Hz), 1.58–1.72 (4H, m), 1.82–1.93 (4H, m), 2.63 (2H, t, J = 7.6 Hz), 2.79 (2H, t, J = 7.6 Hz), 2.97 (3H, s), 3.62 (2H, t, J = 6.5 Hz), 3.84 (2H, t, J = 5.1 Hz), 3.88 (1H, q, J = 6.8 Hz), 6.68 (1H, s), 7.38 (1H, s); MS (EI) m/z 496 [M]+.
1,5,5-Trimethyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)imidazolidine- 2,4-dione (49):
Compound 49 was prepared from compound 39 and 1,5,5-trimethylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.97 (3H, t, J = 7.3 Hz), 0.99 (3H, t, J = 7.3 Hz), 1.39 (6H, s), 1.60–1.67 (4H, m), 1.82–1.92 (4H, m), 2.63 (2H, t, J = 7.6 Hz), 2.78 (2H, t, J = 7.6 Hz), 2.91 (3H, s), 3.62 (2H, t, J = 6.6 Hz), 3.84 (2H, t, J = 6.1 Hz), 6.69 (1H, s), 7.37 (1H, s); MS (EI) m/z 510 [M]+.
5-Methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)imidazolidine-2,4-dione (50):
Compound 50 was prepared from compound 39 and 5-methylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.98 (3H, t, J = 7.3 Hz), 1.00 (3H, t, J = 7.3 Hz), 1.48 (3H, d, J = 6.8 Hz), 1.55–1.75 (4H, m), 1.80–2.00 (4H, m), 2.63 (2H, t, J = 7.8 Hz), 2.79 (2H, t, J = 7.8 Hz), 3.63 (2H, t, J = 6.3 Hz), 3.84 (2H, t, J = 5.9 Hz), 4.12 (1H, q, J = 6.8 Hz), 5.66 (1H, s), 6.69 (1H, s), 7.38 (1H, s); MS (EI) m/z 482 [M]+.
118
1,5-Dimethyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)-5-phenyl- imidazolidine-2,4-dione (51):
Compound 51 was prepared from compound 39 and 1,5-dimethyl-5-phenylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.96 (3H, t, J = 7.3 Hz), 0.98 (3H, t, J = 7.3 Hz), 1.56–1.67 (4H, m), 1.80–1.91 (4H, m), 1.83 (3H, s), 2.61 (2H, t, J = 7.6 Hz), 2.77 (2H, t, J = 7.6 Hz), 2.85 (3H, s), 3.66 (2H, t, J = 6.8 Hz), 3.82 (2H, t, J = 5.9 Hz), 6.68 (1H, s), 7.29–7.44 (6H, m); MS (EI) m/z 572 [M]+.
5-Methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)-5-phenylimidazolidine- 2,4-dione (52):
Compound 52 was prepared from compound 39 and 5-methyl-5-phenylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.3 Hz), 0.97 (3H, t, J = 7.3 Hz), 1.58–1.65 (4H, m), 1.80–1.94 (7H, m), 2.60 (2H, t, J = 7.6 Hz), 2.76 (2H, t, J = 7.8 Hz), 6.23 (1H, brs), 7.32–7.42 (3H, m), 7.50–7.52 (2H, m); MS (EI) m/z 558 [M]+.
5-Methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)-5-p-tolylimidazolidine- 2,4-dione (53):
Compound 53 was prepared from compounds 39 and 42b in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.3 Hz), 0.97 (3H, t, J = 7.3 Hz), 1.58–1.69 (4H, m), 1.75–1.90 (4H, m), 1.83 (3H, s), 2.33 (3H, s), 2.60 (2H, t, J = 7.6 Hz), 2.76 (2H, t, J = 7.6 Hz), 3.62 (2H, t, J = 6.5 Hz), 3.80 (2H, t, J = 7.6 Hz), 6.47 (1H, brs), 6.68 (1H, s), 7.19 (2H, d, J = 8.1 Hz), 7.37 (1H, s), 7.39 (2H, d, J = 8.1 Hz); MS (EI) m/z 572 [M]+.
119
5-(2-Methoxyphenyl)-5-methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (54):
54 was prepared from 39 and 42c in a manner similar to that described for 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.97 (3H, t, J = 7.3 Hz), 0.99 (3H, t, J = 7.6 Hz), 1.59–1.69 (4H, m), 1.78 (3H, s), 1.85–1.98 (4H, m), 2.63 (2H, t, J = 7.6 Hz), 2.79 (2H, t, J = 7.8 Hz), 3.69 (2H, t, J = 6.8 Hz), 3.85 (2H, t, J = 6.1 Hz), 3.88 (3H, s), 6.36 (1H, s), 6.69 (1H, s), 6.94 (1H, d, J = 7.1 Hz), 6.95 (1H, dd, J = 7.6, 8.0 Hz), 7.32 (1H, dd, J = 7.1, 7.6 Hz), 7.38 (1H, s), 7.52 (1H, d, J = 8.0 Hz); MS (EI) m/z 588 [M]+.
5-(3-Methoxyphenyl)-5-methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (55):
Compound 55 was prepared from compounds 39 and 42d in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.3 Hz), 0.97 (3H, t, J = 7.3 Hz), 1.57–1.67 (4H, m), 1.81–1.92 (4H, m), 1.84 (3H, s), 2.60 (2H, t, J = 7.6 Hz), 2.76 (2H, t, J = 7.8 Hz), 3.63 (2H, t, J = 7.1 Hz), 3.81 (2H, t, J = 6.8 Hz), 3.81 (3H, s), 6.21 (1H, brs), 6.68 (1H, s), 6.87 (1H, d, J
= 8.0 Hz), 7.06 (1H, s), 7.08 (1H, d, J = 7.8 Hz), 7.31 (1H, dd, J = 7.8, 8.0 Hz), 7.37 (1H, s); MS (EI) m/z 588 [M]+.
5-(4-Methoxyphenyl)-5-methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (56):
Compound 56 was prepared from compounds 39 and 42a in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.6 Hz), 0.97 (3H, t, J = 7.3 Hz), 1.56–1.69 (4H, m), 1.80–1.89 (4H, m), 1.82 (3H, s), 2.60 (2H, t, J = 7.6 Hz), 2.76 (2H, t, J = 7.6 Hz), 3.63 (2H, t, J = 6.8 Hz), 3.80 (3H, s), 3.81 (2H, t, J = 5.7 Hz), 5.84 (1H, s), 6.68 (1H, s), 6.90 (2H, d, J = 8.9 Hz), 7.36 (1H, s), 7.40 (2H, d, J = 8.9 Hz); MS (EI) m/z 588 [M]+.
120
5-(3,4-Dimethoxyphenyl)-5-methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)- butyl)imidazolidine-2,4-dione (57):
Compound 57 was prepared from compounds 39 and 42e in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.6 Hz), 0.97 (3H, t, J = 7.3 Hz), 1.57–1.68 (4H, m), 1.76–1.90 (4H, m), 1.83 (3H, s), 2.60 (2H, t, J = 7.6 Hz), 2.76 (2H, t, J = 7.8 Hz), 3.64 (2H, t, J = 6.1 Hz), 3.82 (2H, t, J = 6.1 Hz), 3.84 (3H, s), 3.86 (3H, s), 6.27 (1H, brs), 6.68 (1H, s), 6.85 (1H, d, J = 8.8 Hz), 7.03 (1H, s), 7.04 (1H, d, J = 8.8 Hz), 7.37 (1H, s); MS (EI) m/z 618 [M]+.
5-(Benzo[d][1,3]dioxol-5-yl)-5-methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)- butyl)imidazolidine-2,4-dione (58):
Compound 58 was prepared from compounds 39 and 42f in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.96–0.99 (6H, m), 1.57–1.68 (4H, m), 1.81–1.89 (7H, m), 2.61 (2H, t, J = 7.8 Hz), 2.76 (2H, t, J = 7.8 Hz), 3.63 (2H, t, J = 6.7 Hz), 3.82 (2H, t, J = 5.9 Hz), 5.96 (2H, s), 6.51 (1H, brs), 6.68 (1H, s), 6.78 (1H, d, J = 8.1 Hz), 6.95–6.98 (2H, m), 7.37 (1H, s);
MS (EI) m/z 602 [M]+.
5-(4-(Dimethylamino)phenyl)-5-methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)- butyl)imidazolidine-2,4-dione (59):
Compound 59 was prepared from compounds 39 and 42g in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.96 (3H, t, J = 7.1 Hz), 0.97 (3H, t, J = 7.1 Hz), 1.57–1.68 (4H, m), 1.80 (3H, s), 1.80–1.89 (4H, m), 2.61 (2H, t, J = 7.6 Hz), 2.76 (2H, t, J = 7.8 Hz), 2.93 (6H, s), 3.62 (2H, t, J = 6.6 Hz), 3.81 (2H, t, J = 6.1 Hz), 6.21 (1H, s), 6.68 (1H, s), 6.69 (2H, d, J = 9.0 Hz), 7.32 (2H, d, J = 9.0 Hz), 7.37 (1H, s); MS (EI) m/z 601 [M]+.
121
5-Methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)-5-(4-(trifluoromethyl)- phenyl)imidazolidine-2,4-dione (60):
Compound 60 was prepared from compounds 39 and 42h in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.92–0.98 (6H, m), 1.57–1.67 (4H, m), 1.80–1.90 (7H, m), 2.59 (2H, t, J = 7.7 Hz), 2.76 (2H, t, J = 7.8 Hz), 3.64 (2H, t, J = 7.0 Hz), 3.81 (2H, t, J = 5.9 Hz), 6.63 (1H, s), 6.69 (1H, s), 7.37 (1H, s), 7.64–7.71 (4H, m); MS (EI) m/z 626 [M]+.
5-Methyl-5-(4-nitrophenyl)-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (61):
Compound 61 was prepared from compounds 39 and 42i in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.94 (3H, t, J = 7.3 Hz), 0.96 (3H, t, J = 7.1 Hz), 1.57–1.67 (4H, m), 1.81–1.93 (4H, m), 1.90 (3H, s), 2.60 (2H, t, J = 7.8 Hz), 2.76 (2H, t, J = 7.8 Hz), 3.66 (2H, t, J = 6.8 Hz), 3.82 (2H, t, J = 5.6 Hz), 6.69 (1H, s), 6.88 (1H, s), 7.37 (1H, s), 7.78 (2H, d, J = 8.8 Hz), 8.24 (2H, d, J = 8.8 Hz); MS (EI) m/z 603 [M]+.
tert-Butyl 4-(4-methyl-2,5-dioxo-1-(4-((2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yl)oxy)butyl)- imidazolidin-4-yl)benzoate:
The title compound was prepared from compounds 39 and 42j in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.94 (3H, t, J = 7.3 Hz), 0.96 (3H, t, J = 7.3 Hz), 1.51–1.70 (13H, m), 1.79–1.92 (7H, m), 2.60 (2H, t, J = 7.8 Hz), 2.76 (2H, t, J = 7.8 Hz), 3.64 (2H, t, J = 6.8 Hz), 3.81 (2H, t, J = 6.1 Hz), 6.68 (1H, s), 6.79 (1H, brs), 7.37 (1H, s), 7.60 (2H, d, J = 8.8 Hz), 8.00 (2H, d, J = 8.8 Hz); MS (EI) m/z 658 [M]+.
122
4-(4-Methyl-2,5-dioxo-1-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)imidazolidin -4-yl)benzoic acid (62):
To a solution of the precucor (30 mg, 46 mol) in CH2Cl2 (1 mL), TFA (1 mL) was added dropwise at 0 °C. The reaction mixture was stirred at room temperature for 30 min. The reaction mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc = 1/1) to give the title compound (25.9 mg, 94%) as a white solid; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.3 Hz), 0.96 (3H, t, J = 7.3 Hz), 1.56–1.67 (4H, m), 1.81–1.90 (4H, m), 1.88 (3H, s), 2.60 (2H, t, J = 7.6 Hz), 2.75 (2H, t, J = 7.8 Hz), 3.69 (2H, t, J = 7.3 Hz), 3.82 (2H, t, J = 7.1 Hz), 6.71 (1H, s), 7.39 (1H, s), 7.45 (1H, s), 7.65 (2H, d, J = 8.5 Hz), 8.12 (2H, d, J = 8.5 Hz), 9.79 (1H, brs); MS (EI) m/z 602 [M]+.
5-Methyl-3-(4-((2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yl)oxy)butyl)-5-(4-((tetrahydro-2H- pyran-2-yl)oxy)phenyl)imidazolidine-2,4-dione:
The title compound was prepared from compounds 39 and 42k in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.3 Hz), 0.97 (3H, t, J = 7.3 Hz), 1.57–1.73 (7H, m), 1.82–2.01 (10H, m), 2.61 (2H, t, J = 7.8 Hz), 2.77 (2H, t, J = 7.8 Hz), 3.56–3.64 (3H, m), 3.80–3.88 (3H, m), 5.40 (2H, s), 6.07 (1H, s), 6.68 (1H, s), 7.05 (2H, d, J = 8.8 Hz), 7.37 (1H, s), 7.39 (2H, d, J = 8.8 Hz); MS (EI) m/z 658 [M]+.
5-(4-Hydroxyphenyl)-5-methyl-3-(4-(2-oxo-6,8-dipropyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (63):
To a solution of the precucor (30 mg, 46 mol) in THF (570 L) and water (285 L), AcOH (1.1 mL) was added at 0 °C. The reaction mixture was stirred at room temperature for 6 h. The reaction mixture was diluted with saturated NaHCO3 aq. at 0 °C and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/EtOAc
= 1/1) to give the title compound (23.5 mg, 90%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.87–0.97 (6H, m), 1.53–1.67 (4H, m), 1.81–1.88 (7H, m), 2.60 (2H, t, J = 7.7 Hz), 2.71 (2H, t, J = 7.8 Hz), 3.63 (2H, t, J = 6.8 Hz), 3.80–3.86 (2H, m), 4.91 (1H, s), 6.41 (1H, s), 6.69 (1H, s), 6.78–6.80 (2H, m), 7.28–7.31 (2H, m), 7.37 (1H, s); MS (EI) m/z 574 [M]+.
123 Synthesis of the 2-oxochromen derivative with propyl group
Compounds 69, 70 and 64~68 were prepared in the same manner as the synthesis of 2.
Characterization data
The NMR and MS data of compounds 69, 70 and 64~68 are described below.
1-Methyl-3-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)imidazolidine-2,4-dione (64):
Compound 64 was prepared from compound 40 and 1-methylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.95 (3H, t, J = 7.3 Hz), 1.58 (2H, qt, J = 7.3, 7.6 Hz), 1.86–1.87 (4H, m), 2.82 (2H, t, J = 7.6 Hz), 3.01 (3H, s), 3.61 (2H, t, J = 3.9 Hz), 3.88 (2H, s), 4.11 (2H, t, J = 4.3 Hz), 6.60 (1H, s), 6.88 (1H, d, J = 9.0 Hz), 7.54 (1H, d, J
= 9.0 Hz); MS (EI) m/z 440 [M]+.
1,5,5-Trimethyl-3-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)imidazolidine-2,4- dione (65):
Compound 65 was prepared from compound 40 and 1,5,5-trimethylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.96 (3H, t, J = 7.3 Hz), 1.39 (6H, s), 1.55–1.61 (2H, m), 1.86 (4H, brs), 2.83 (2H, t, J = 7.6 Hz), 2.90 (3H, s), 3.57–3.63 (2H, m), 4.10–4.13 (2H, m), 6.61 (1H, s), 6.88 (1H, d, J = 9.0 Hz), 7.54 (1H, d, J = 8.9 Hz); MS (EI) m/z 468 [M]+.
5-Methyl-3-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)-5-phenylimidazolidine- 2,4-dione (66):
Compound 66 was prepared from compound 40 and 5-methyl-5-phenylimidazolidine-2,4-dione in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.92 (3H, t, J = 7.3 Hz), 1.55 (2H, qt, J = 7.3, 7.3 Hz), 1.76–1.86 (7H, m), 2.79 (2H, t, J = 7.3 Hz), 3.61
124
(2H, t, J = 6.2 Hz), 4.07 (2H, t, J = 5.1 Hz), 6.60 (1H, s), 6.83 (1H, d, J = 9.2 Hz), 7.30–7.42 (3H, m), 7.49–7.53 (3H, m); MS (EI) m/z 516 [M]+.
5-(3,4-Dimethoxyphenyl)-5-methyl-3-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (67):
Compound 67 was prepared from compounds 40 and 42e in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.93 (3H, t, J = 7.3 Hz), 1.51–1.60 (2H, m), 1.82–1.98 (7H, m), 2.80 (2H, t, J = 7.6 Hz), 3.61 (2H, t, J = 6.2 Hz), 3.86 (3H, s), 3.89 (3H, s), 4.04–4.09 (2H, m), 6.24 (1H, s), 6.61 (1H, s), 6.83–6.86 (2H, m), 7.02–7.04 (2H, m), 7.50–7.53 (1H, m); MS (EI) m/z 576 [M]+.
5-(Benzo[d][1,3]dioxol-5-yl)-5-methyl-3-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (68):
Compound 68 was prepared from compounds 40 and 42f in a manner similar to that described for compound 2.
The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.93 (3H, t, J = 7.3 Hz), 1.51–1.63 (2H, m), 1.79–1.85 (7H, m), 2.80 (2H, t, J = 7.6 Hz), 3.61 (2H, t, J = 6.1 Hz), 4.06–4.09 (2H, m), 5.96 (2H, s), 6.34 (1H, brs), 6.61 (1H, s), 6.78 (1H, d, J = 8.3 Hz), 6.85 (1H, d, J = 9.0 Hz), 6.95 (1H, dd, J = 2.0, 8.3 Hz), 6.99 (1H, d, J = 2.0 Hz), 7.53 (1H, d, J = 9.0 Hz); MS (EI) m/z 560 [M]+.
tert-Butyl 4-(4-methyl-2,5-dioxo-1-(4-((2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yl)oxy)butyl)- imidazolidin-4-yl)benzoate:
The title compound was prepared from compounds 40 and 42j in a manner similar to that described for compound 2. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.92 (3H, t, J = 7.3 Hz), 1.50–1.62 (11H, m), 1.82–1.89 (7H, m), 2.80 (2H, t, J = 7.8 Hz), 3.62 (2H, t, J = 6.6 Hz), 4.07 (2H, t, J = 5.6 Hz), 6.60 (1H, s), 6.64 (1H, brs), 6.85 (1H, d, J = 9.0 Hz), 7.53 (1H, d, J = 9.0 Hz), 7.58 (2H, d, J = 8.3 Hz), 8.00 (2H, d, J = 8.3 Hz); MS (EI) m/z 616 [M]+.
125
4-(4-Methyl-2,5-dioxo-1-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)imidazolidin- 4-yl)benzoic acid (69):
Compound 69 was prepared from the precucor in a manner similar to that described for compound 62. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.91 (3H, t, J = 7.3 Hz), 1.54 (2H, qt, J
= 7.3, 7.6 Hz), 1.84–1.93 (7H, m), 2.78 (2H, t, J = 7.6 Hz), 3.66 (2H, t, J = 6.5 Hz), 4.08 (2H, t, J = 5.6 Hz), 6.64 (1H, s), 6.86 (1H, d, J = 9.0 Hz), 7.54–7.56 (2H, m), 7.63 (2H, d, J = 8.6 Hz), 8.11 (2H, d, J = 8.6 Hz), 10.91 (1H, brs); MS (EI) m/z 560 [M]+.
5-(4-Hydroxyphenyl)-5-methyl-3-(4-(2-oxo-8-propyl-4-(trifluoromethyl)-2H-chromen-7-yloxy)butyl)- imidazolidine-2,4-dione (70):
Compound 70 was prepared in a manner similar to that described for compound 63. The title compound was obtained as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 0.89 (3H, t, J = 7.3 Hz), 1.48–1.57 (2H, m), 1.79 (3H, s), 1.79–1.89 (4H, m), 2.76 (2H, t, J = 7.3 Hz), 3.60 (2H, t, J = 5.8 Hz), 4.07 (2H, t, J = 5.1 Hz), 6.54 (1H, s), 6.60 (1H, s), 6.78 (2H, d, J = 7.6 Hz), 6.84 (1H, d, J = 9.0 Hz), 7.28 (2H, d, J = 7.6 Hz), 7.52 (1H, d, J = 9.0 Hz); MS (EI) m/z 532 [M]+.
Pharmacological Experimental Details
Material and Method
In vitro study
Measurement of ABCA1 and SREBP-1c mRNA levels
The THP-1 and HepG1 cells were stably seeded at semi-confluent growth on a 24-well plate in CSC medium supplemented with 10% bovine fetal serum and incubated under a wet atmosphere with 5% CO2 at 37 C. After 24 h of incubation the medium with 5 M concentration of the test compound was added, and the cells were further incubated for 24 h. Next, RNA was extracted using Isogen (Japan Gene Inc.) to produce cDNA with Randam Primer (ABI Inc.) as a substrate, and the relative expression levels of ABCA1 and SREBP-1c mRNA were measured with real-time quantitative PCR to determine the effect of the test compound.
126 GAL4-h-LXRs reporter gene assay
The CHO K-1 cells stably transfected with LXRs/GAL4-fused protein expression vectors and the GAL4-responsive reporter vector (pG5luc; Promega, WI) were seeded at 20,000 cells/well on a 96-well plate in HAM-F12 medium supplemented with 10% bovine fetal serum, 100 units/mL of penicillin G, and 100 g/mL of streptomycin sulfate, followed by incubation under a wet atmosphere with 5% CO2 at 37 C. After 24 h, media with different concentrations of the test compound (0.01, 0.1, 1, and 10 M) were added, and the cells were further incubated for 24 h. Bright-Glo (Promega) was used as a substrate, and the luminescence intensity was measured by using the LB960 Luminometer (Berthold Technologies) to dertermine the effect of the test compound on the activation of luciferase transcription via LXR- or LXR-LBD.
In vivo study
Bio F1B hamsters (male, age: 8 weeks old, n = 42) (Charles River Japan, Inc., Kanagawa, Japan) were used for animal experiments. The animal room was controlled at 23 ± 3 °C and relative humidity of 50 ± 20%. Animals were fed a CE-2 chow diet (CLEA Japan Inc., Tokyo, Japan) and then supplemented with CE-2 containing 0.3%
cholesterol and 10% coconut oil for 10 weeks.
During fat loading, compound 2 was orally administered at 30, 100, or 300 mg/kg/day (n = 6 for each dose group).
The comparative agent, T0901317 was orally administered at 1, 3, or 10 mg/kg/day (n = 6 for each dose group).
The control group (n = 12) received an aqueous solution of 0.5% methylcellulose instead of 2. Blood was sampled for determination of plasma lipid levels by using a commercial kit (total cholesterol: Cholesterol E-Test Wako;
Wako Pure Chemical Industries, Osaka, Japan; triglyceride: Triglyceride E-Test Wako; Wako Pure Chemical Industries). The plasma lipoprotein profiles were analyzed by using the CLiP method 80) on the LC-20A HPLC System (Shimadzu) and Superose 6 Column (10 mm×300 mm, GE Healthcare, UK). Briefly, 15 L of hamster plasma was 10-fold diluted with PBS containing 1 mM EDTA. Diluted plasma (20 L) was separated by the column at 0.5 mL/min with the same buffer maintained at 40 °C for the simultaneous determination of cholesterol contents in the eluents.
For the analyses of atherosclerotic lesions, the animals were anesthetized via intraperitoneal injection of pentobarbital sodium (50 mg/kg), followed by vascular perfusion for 5 min with saline containing 4%
paraformaldehyde with a perfusion pressure of 120 mm H2O.
The aorta was separated from the heart at the 15 mm upper portion from the aortic origin. The isolated thoracic aorta was cut open, mounted on a rubber plate, and fixed with 4% paraformaldehyde followed by staining with Oil-Red O. Fatty streak areas were measured with an image analysis system (SP500F; Olympus, Tokyo, Japan).
127 Pharmacokinetic Experimental Details
Material and Method Pharmacokinetic studies
A solution of compound 2 in PEG400 was orally administered to Golden Syrian hamsters at a dose of 300 mg/kg, and their blood samples were collected from the jugular vein at 0.5, 1, 2, and 6 h after dosage and immediately centrifuged to obtain the corresponding plasma fractions. The Oasis HLB-cartridge (1 cc, 30 mg) was preconditioned by eluting MeOH (1 mL) and then rinsed with deionized water (1 mL). The plasma samples (100
L) and the corresponding internal standards (50 L) were passed through the HLB-cartridge. The cartridge was washed with 5% aqueous MeOH (1 mL), and subsequently the drug residues were eluted from the cartridge with MeOH (2 mL). The extracts were completely evaporated to dryness by nitrogen stream. The residues were then dissolved in MeCN (75 L) and 1% aqueous AcOH (75 L). The drug concentration in the solution was measured by the LC-10 HPLC System (Shimadzu). Compound 2 showed a low concentration in the plasma for up to 2 h after administration and disappeared 6 h after administration, as shown in the drug concentration curve obtained for a dose of 300 mg/kg (Figure 24).
Molecular Modeling
Methods
1) Receptor structure preparation.
The X-ray crystal structure of human LXRβ/GW3965 complex (Protein Data Bank [PDB] ID: 1PQ6) was downloaded from the PDB. The raw PDB and our X-ray crystal structures (data not shown) were converted into an all-atom, fully prepared receptor model structure by using the Protein Preparation Wizard in the Maestro 42b). These structures were superimposed with the Molecular Operating Environment (MOE) 42c). The docking receptor grids were created by the Glide’s Receptor Grid Generation module 42b).
2) Ligand preparation.
All ligands were processed with the LigPrep 42b) applying the following conditions: generation of tautomers, generation of stereoisomers and ionization at pH = 7 with a tolerance of ± 2.
3) Ligand-protein docking.
Standard precision mode of the Glide 42b) program was used for all docking calculations. The van der Waals radii of ligands were scaled by 0.8. During the initial phase of docking calculation, the maximum poses generated from the variables were fixed to 5,000 and the best variable that set the number of poses per ligand that enters the energy minimization was set to 1,000. The final docking model was selected by comparing with our X-ray crystal structure.