The particularity of the marine environment leads to the rich bromine in the marine natural products, so the syntheses of bromo-substituted indole compounds are the key step in the preparations of marine alkaloid Eudistomins Y₁–Y₇ 23–29. The target compounds were started from the commercially available indole. Substituted indole-3-carboxaldehyde 2a–c was synthesized from 1a–c by Vilsmeier-Hacck reaction with phosphorus oxychloride (POCl₃) and N,N-dimethylformamide (DMF) according to the reference [8]. Intermediates 2a–c was condensed with nitromethane (CH₃NO₂) based on the classical Henry reaction to form the corresponding vinyl nitro compounds 3a–c [9], reduction of which by NaBH₄ at room temperature gave compounds 4a–c in high yields. Subsequently, 4a was reduced by Pd/C in methanol at room temperature to give 5a. 4b–c were reduced by LiAlH₄ and refluxed in THF to give bromo-substituted tryptamines 5b–c [10]. The reaction routes were outlined in Scheme 1.

Methylation of bromophenol by dimethyl sulfate in acetone, 7 and 12 was converted to substituted methoxybenzene 8 and 13. Intermediate 14 was prepared by 1-bromo-2-methoxybenzene 13 using acetyl chloride by Friedel–Craft acylation. The substituted acetophenones 8, 10 and 14 were converted to substituted phenylglyoxals 9, 11 and 15 by oxidation with SeO₂ in dioxane–water mixture [11]. The reaction routes were outlined in Scheme 2.

The classical method to prepare β-carboline alkaloids through Pictet-Spengler reaction was a two-step method, and involved the acid-catalyzed condensation of an aliphatic amine attached to a sufficiently reactive aromatic nucleus with aldehydes. Specifically, in the first step, an imine was formed, which can be activated by acids, while in the second step, the endo cyclization occurred between a carbon nucleophile of a sufficiently reactive aromatic moiety and the activated iminium ion, resulting in tetrahydro-β-carboline by the formation of N-heterocyclic ring through a new C–C bond. After dehydrogenation, tetrahydro-β-carboline was converted to β-carboline [12,13,14,15]. In our experiments, the treatment of substituted tryptamines with substituted phenylglyoxal [16,17] under the acidic conditions did not produce the expected tetrahydro-β-carboline but rather directly generated a dehydrogenated β-carboline product 16–22 as reported by Zhang et al. [18]. It is possible that the acidity and polarity of glacial acetic acid promote the reaction of cyclization and dehydrogenation aromatizing to completion continuously. The reaction routes were outlined in Scheme 3. Pictet-Spengler cyclization of substituted tryptamine 5a–d with substituted phenylglyoxal 9, 11 or 15 in acetic acid afforded compounds 16–22, which was transformed into the target compounds Eudistomins Y₁–Y₇ 23–29 in the presence of BBr₃ in CH₂Cl₂ at −1278 °C with the yields of 62%–68%. The reaction routes were outlined in Scheme 3.

The modified one-pot oxidation reaction shown as the Scheme 3 was more efficient and convenient in preparing 1-substituted β-carbolines without the need of aromatization step or decarboxylation. Using this modified Pictet-Spengler reaction, the target compounds Eudistomins Y₁–Y₇ 23–29 were synthesized.

The in vitro anti-proliferative activity of select compounds 16–29 was evaluated against the breast carcinoma cell line MDA-231 using the 3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyltetrazolium bromide (MTT) metabolic assay. Briefly, exponentially growing cells (1 × 10³ cells) were seeded in 96-well plates. After 18 h, cells were continuously treated with compounds 16–29. Following this, after 96 h, cell survival was evaluated. The inhibitory activity (IC₅₀) of the various compounds on cell proliferation was determined (Table 1). Moderate anti-proliferative activity was observed with all the compounds tested. Surprisingly, Eudistomins Y₁–Y₃ and Y₅–Y₇ with a hydroxyl group were found to have a poor cytotoxic activity compared with their precursors 16–18 and 20–22. In summary, in vitro inhibitory activities of methylated products were preferable to the demethylated products except for compounds 19 and 26.

In order to determine the intracellular target of these compounds, the inhibitory activity of compound 26 was tested against various kinases in vitro. Briefly, human recombinant full-length kinases were incubated in kinase buffer containing ATP and substrate (Poly Glu:Tyr) for 4 h at room temperature with or without the presence of compounds 26 at 10 μM final concentration. Remaining ATP in solution was then quantified utilizing the Kinase-Glo-luminescence kit (Promega). The ability of compound 26 to inhibit the kinase activity of these kinases was evaluated (Table 2). As such, the anti-proliferative activity of compound 26 does not seem to be significantly modulated by any of the kinases tested.

The starting materials and reagents, purchased from commercial suppliers, were used without further purification. All reactions were monitored by thin-layer chromatography (TLC), on aluminium sheets (Silica gel 60-F₂₅₄, E. Merck). Compounds were visualized by UV light. Column chromatography was carried out using silica gel (200–300 mesh). All reaction solvents were dried prior to use according to standard procedures. All primary reagents were commercially available. Silica gel chromatography solvents were of analytical grade. Melting points were recorded on a micro melting point apparatus MP-500D and were uncorrected. NMR spectra were recorded on a Jeol JNM-ECP spectrometer at 600 MHz for ¹H NMR and 150 MHz for ¹³C NMR with TMS as the internal standard. Chemical shifts are expressed in δ (ppm) and coupling constants (J) in Hz. Multiplicity is indicated as follows: s (singlet), d (doublet), t (triplet), dd (doublet of doublets), brs (broad singlet), etc. Mass spectra were recorded using a Q-TOF Ultima™ Global by chemical ionization.

In a separate tear-shaped flask under a nitrogen atmosphere, 40 mL of DMF was cooled to 0 °C, and then POCl₃ (150 mmol) was added dropwise within 0.5 h under nitrogen. After the addition, stirring was continued for 1.5 h at 0 °C. It was followed by addition of a solution of indole 1a–c (100 mmol) in DMF (20 mL) below 0 °C within 1.5 h. Then the reaction solution was heated to 40 °C for another 2 h and cooled to 0 °C. Subsequently, crushed ice (50 g) was added, and then the pH was adjusted to 8–9 by adding NaOH solution (20%, w/w). The resulting reaction mixture was then heated at reflux for 12 h. And then, the resulting suspension solution was cooled naturally to the room temperature and filtered through celite to afford a white crude product. The crude product was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:3) as the mobile phase to provide the compounds 2a–c [8].

1H-Indole-3-carbaldehyde (2a): White solid; yield 96%; ¹H NMR (600 MHz, CDCl₃) δ 11.21 (brs, 1H), 10.04 (s, 1H), 8.24 (d, J = 7.8 Hz, 1H), 8.21 (s, 1H), 7.54 (d, J = 7.8 Hz, 1H), 7.30–7.23 (m, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 184.6, 137.3, 124.7, 123.7, 122.2, 121.4, 119.2, 112.2.

6-Bromo-1H-indole-3-carbaldehyde (2b): White solid; yield 95%; ¹H NMR (600 MHz, CDCl₃) δ 11.33 (brs, 1H), 10.03 (s, 1H), 8.24 (s, 1H), 8.15 (d, J = 8.3 Hz, 1H), 7.76 (d, J = 1.8 Hz, 1H), 7.39 (dd, J = 8.3, 1.8 Hz, 1H); ¹³C NMR (151 MHz, CDCl₃) δ 184.7, 137.9, 125.3, 123.6, 122.9, 119.0, 116.5, 115.2.

5-Bromo-1H-indole-3-carbaldehyde (2c): White solid; yield 95%; ¹H NMR (600 MHz, CDCl₃) δ 11.36 (brs, 1H), 10.02 (s, 1H), 8.39 (d, J = 1.8 Hz, 1H), 8.26 (s, 1H), 7.53 (d, J = 8.7 Hz, 1H), 7.40 (dd, J = 8.7 Hz, 1.8 Hz, 1H); ¹³C NMR (151 MHz, CDCl₃) δ 184.6, 138.2, 136.1, 126.5, 126.4, 123.8, 118.5, 115.3, 114.2.

To a solution of 1H-indole-3-carbaldehyde 2a–c (80 mmol) in CH₃NO₂ (80 mL), was added ammonium acetate (40 mmol) and benzene (1.1 mL, 12.4 mmol). The reaction mixture was heated at reflux for 12 h. After cooling to room temperature, some solvent was removed under reduced pressure and filtered through Celite to afford a yellow crude product. The crude product was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:2) as the mobile phase to provide the compounds 3a–c [9].

(E)-3-(2-Nitrovinyl)-1H-indole (3a): Deep yellow solid; yield 92%; ¹H NMR (600 MHz, CDCl₃) δ 11.33 (brs, 1H), 8.39 (d, J = 13.3 Hz, 1H), 8.16 (d, J = 3.2 Hz, 1H), 7.97 (dd, J = 5.9, 2.3 Hz, 1H), 7.92 (d, J = 13.3 Hz, 1H), 7.59 (dd, J = 5.9, 2.3 Hz, 1H), 7.32–7.29 (m, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 138.2, 135.1, 133.9, 131.9, 125.1, 123.6, 122.1, 120.5, 112.8, 108.8.

(E)-6-Bromo-3-(2-nitrovinyl)-1H-indole (3b): Deep yellow solid; yield 91%; ¹H NMR (600 MHz, CDCl₃) δ 11.39 (brs, 1H), 8.36 (d, J = 13.3 Hz, 1H), 8.18 (s, 1H), 7.95 (d, J = 8.3 Hz, 1H), 7.91 (d, J = 13.3 Hz, 1H), 7.78 (d, J = 1.8 Hz, 1H), 7.41 (dd, J = 8.3, 1.8 Hz, 1H); ¹³C NMR (151 MHz, CDCl₃) δ 138.9, 135.4, 133.2, 132.7, 124.9, 124.1, 122.0, 116.5, 115.7, 108.8.

(E)-5-Bromo-3-(2-nitrovinyl)-1H-indole (3c): Deep yellow solid; yield 91%; ¹H NMR (600 MHz, CDCl₃) δ 11.41 (brs, 1H), 8.35 (d, J = 13.7 Hz, 1H), 8.19 (s, 1H), 8.16 (d, J = 1.8 Hz, 1H), 7.97 (d, J = 13.7 Hz, 1H), 7.54 (d, J = 8.7 Hz, 1H), 7.42 (dd, J = 8.7, 1.8 Hz, 1H); ¹³C NMR (151 MHz, CDCl₃) δ 136.7, 135.5, 133.0, 132.7, 126.8, 126.3, 122.9, 115.0, 114.5, 108.3.

To a solution of (E)-3-(2-nitrovinyl)-1H-indole 3a–c (20mmol) in THF (60 mL) and CH₃OH (9 mL), was added NaBH₄ (40 mmol) in batch over 0.5 h. The above reaction solution was stirred at room temperature for about 1 h and the completion of the reaction was monitored by TLC. Then water (100 mL) and hydrochloric acid (100 mL, 10%, v/v) was added slowly. The resulting reaction mixture was extracted with CH₂Cl₂ (30 mL × 3). The combined organic phase was washed with H₂O (20 mL × 3) and brine (20 mL × 3), dried over anhydrous MgSO₄ and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:3) as the mobile phase to provide the compounds 4a–c [10].

3-(2-Nitroethyl)-1H-indole (4a): Brown solid; yield 96%; ¹H NMR (600 MHz, CDCl₃) δ 8.09 (brs, 1H), 7.59 (d, J = 7.8 Hz, 1H), 7.38 (d, J = 8.3 Hz, 1H), 7.25 (t, J = 7.8, 7.3 Hz, 1H), 7.18 (t, J = 8.3, 7.3 Hz, 1H), 7.04 (s, 1H), 4.67 (t, J = 6.9 Hz, 2H), 3.49 (t, J = 6.9 Hz, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 136.3, 126.7, 122.7, 122.6, 119.9, 118.2, 111.6, 110.0, 75.8, 23.7.

6-Bromo-3-(2-nitroethyl)-1H-indole (4b): Brown solid; yield 98%; ¹H NMR (600 MHz, CDCl₃) δ 8.13 (brs, 1H), 7.49 (d, J = 1.8 Hz, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.24 (dd, J = 8.2, 1.8 Hz, 1H), 7.00 (d, J = 1.8 Hz, 1H), 4.64 (t, J = 6.9 Hz, 2H), 3.49 (t, J = 6.9 Hz, 2H); ¹³C NMR (125 MHz, CDCl₃) δ 137.1, 125.7, 123.3, 123.2, 119.5, 116.1, 114.5, 110.3, 75.8, 23.5.

5-Bromo-3-(2-nitroethyl)-1H-indole (4c): Brown solid; yield 95%; ¹H NMR (600 MHz, CDCl₃) δ 8.18 (brs, 1H), 7.68 (d, J = 1.8 Hz, 1H), 7.29 (dd, J = 8.7, 1.8 Hz, 1H), 7.22 (d, J = 8.7 Hz, 1H), 7.03 (d, J = 1.8 Hz, 1H), 4.64 (t, J = 7.3 Hz, 2H), 3.41 (t, J = 7.3 Hz, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 134.9, 128.5, 125.4, 124.0, 120.8, 113.2, 113.1, 109.7, 75.7, 23.4.

2-(1H-indol-3-yl)ethanamine (5a): To a solution of 3-(2-nitroethyl)-1H-indole 4a (1.9 g, 10 mmol) in methanol (30 mL), was added 10% Pd/C (0.08 g). H₂ was passed in at room temperature, then hydrogenated under normal pressure for 24 h. The completion of the reaction was monitored by TLC. The reaction mixture was then filtered and evaporated under reduced pressure. The residue was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/methanol (10:1) as the mobile phase to provide 2-(1H-indol-3-yl)ethanamine 5a as an white solid in a yield of 98%. ¹H NMR (600 MHz, CDCl₃) δ 11.04 (brs, 1H), 7.57 (d, J = 7.8 Hz, 1H), 7.42 (d, J = 7.8 Hz, 1H), 7.17 (s, 1H), 7.11 (t, J = 7.8 Hz, 1H), 7.02 (t, J = 7.8 Hz, 1H), 2.89 (t, J = 6.8 Hz, 2H), 2.83 (t, J = 6.8 Hz, 2H), 2.70 (s, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 136.9, 127.9, 123.2, 121.4, 118.9, 113.0, 111.9, 43.1, 29.8.

To a solution of substituted 3-(2-nitroethyl)-1H-indole 4b–c (10 mmol) in THF (30 mL), was added LiAlH₄ (20 mmol) slowly. After the addition, it was heated at reflux for about 5 h. The completion of the reaction was monitored by TLC. The mixture was allowed to cool to room temperature and quenched by dropwise addition of saturated Na₂SO₄ solution. The resulting suspension mixture was filtered through Celite, and the filtrate was extracted with EtOAc (30 mL × 3). The combined organic phase was washed with H₂O (20 mL × 3) and brine (20 mL × 3), dried over anhydrous MgSO₄ and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/methanol (10:1) as the mobile phase to provide the compounds 5b–c as a gray solid [10].

2-(6-Bromo-1H-indol-3-yl)ethanamine (5b): White solid; yield 92%; ¹H NMR (600 MHz, CDCl₃) δ 11.08 (brs, 1H), 7.52 (d, J = 1.8 Hz, 1H), 7.47 (d, J = 8.7 Hz, 1H), 7.18 (s, 1H), 7.09 (dd, J = 8.7, 1.8 Hz, 1H), 3.53 (s, 2H), 2.83 (t, J = 6.4 Hz, 2H), 2.77 (t, J = 6.4 Hz, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 137.7, 126.9, 124.4, 121.5, 120.7, 114.5, 114.2, 113.1, 42.6, 28.7.

2-(5-Bromo-1H-indol-3-yl)ethanamine (5c): White solid; yield 88%; ¹H NMR (600 MHz, CDCl₃) δ 11.17 (brs, 1H), 7.75 (d, J = 1.8 Hz, 1H), 7.31–7.21 (m, 2H), 7.16 (s, 1H), 3.51 (s, 2H), 2.83 (t, J = 6.4 Hz, 2H), 2.74 (t, J = 6.4 Hz, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 135.4, 129.7, 124.9, 124.0, 123.7, 121.1, 113.9, 111.2, 42.4, 28.5.

1-(3,5-Dibromo-4-hydroxyphenyl)ethanone (7): To a suspended mixture of 1-(4-hydroxyphenyl)ethanone (4.08 g, 30 mmol) and deionized water (200 mL), was added N-Bromosuccinimide (8.01 g, 45 mmol). The reaction mixture was heated to 60 °C, then 40% (v/v) H₂SO₄ (20 mL) was added. After stirred for about 10 h and the completion of the reaction was monitored by TLC, the reaction mixture was filtered through Celite to afford a white crude product. The crude product was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:3) as the mobile phase to provide a compound 1-(3,5-dibromo-4-hydroxyphenyl)ethanone (7) as a white solid in the yield of 92%.

1-(3,5-Dibromo-4-methoxyphenyl)ethanone (8): To a solution of 7 (20 mmol) in anhydrous acetone (50 mL) were added potassium carbonate (30.0 mmol) and dimethyl sulfate (2.84 mL, 30.0 mmol), and refluxed for 2 h. After cooled to room temperature, the reaction mixture was filtered through Celite, and the filtrate was concentrated under vacuum. The residue was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:2) as the mobile phase to provide the desired compound 8 as a white solid in the yield of 95%. ¹H NMR (600 MHz, CDCl₃) δ 8.08 (s, 2H), 3.93 (s, 3H), 2.56 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 194.5, 158.1, 135.1, 133.0, 118.7, 60.9, 26.6.

To a solution of dioxane/deionized water (20:1) (30 mL) was added SeO₂ (12 mmol). The mixture was heated to 80 °C and stirred until the solid dissolved. It was followed by addition of substituted phenylethanone 8, 10, 14 (10 mmol), and was refluxed for about 12 h and the completion of the reaction was monitored by TLC. The hot solution was filtered through Celite, and the filtrate was concentrated under vacuum. The residue was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:3) as the mobile phase to provide the desired compound 9, 11, 15 [11].

2-(3,5-Dibromo-4-methoxyphenyl)-2-oxoacetaldehyde (9): Light brown solid; yield 71%; ¹H NMR (600 MHz, CDCl₃) δ 9.42 (s, 1H), 8.28 (s, 1H), 8.22 (s, 1H), 3.85 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 193.6, 191.9, 157.9, 134.5, 132.3, 118.3, 61.2.

2-(4-Methoxyphenyl)-2-oxoacetaldehyde (11): Light brown solid; yield 75%; ¹H NMR (600 MHz, CDCl₃) δ 9.53 (s, 1H), 8.06 (d, J = 8.7 Hz, 2H), 7.02 (d, J = 8.7 Hz, 2H), 3.83 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 195.2, 192.5, 164.0, 132.4, 126.7, 114.4, 56.1.

2-(3-Bromo-4-methoxyphenyl)-2-oxoacetaldehyde (15): Light brown solid; yield 74%; ¹H NMR (600 MHz, CDCl₃) δ 9.43 (s, 1H), 8.28 (s, 1H), 8.09 (d, J = 8.7 Hz, 1H), 7.21 (d, J = 8.7 Hz, 1H), 3.93 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 194.4, 192.6, 159.9, 134.8, 131.7, 127.8, 112.7, 110.9, 57.2.

1-(3-Bromo-4-methoxyphenyl)ethanone (14): To a suspension mixture of anhydrous AlCl₃ (3.0 g, 22.5 mmol) in CS₂ (50 mL) , was added 1-bromo-2-methoxybenzene (2.79 g, 15 mmol). The above mixture was maintained at room temperature and stirred for 0.5 h, and then acetyl chloride (1.6 mL, 22.5 mol) was added dropwise. After the addition, stirring was continued for another 0.5 h. And then, the reaction mixture was refluxed for 2 h. After cooled to room temperature, it was poured into 80 mL of ice-water containing 20 mL of concentrated hydrochloric acid and stirred to reach room temperature. The mixture was extracted with CH₂Cl₂ (20 mL × 3). The combined organic phase was washed with H₂O (20 mL × 3), 10% aqueous NaOH (10 mL × 2), H₂O (20 mL × 3) and brine (20 mL × 3), dried over anhydrous MgSO₄ and the solvent was removed under reduced pressure. The residue was purified by recrystallization from petroleum ether (20 mL) to afford 1-(3-bromo-4-methoxyphenyl)ethanone (14) as a brown solid in the yield of 76%.

To a solution of substituted phenylglyoxal 9, 11, 15 (3 mmol) in AcOH (30 mL), was added substituted tryptamine 5a–d (3 mmol). The above mixture was heated at 90 °C for 10 h, then cooled and adjusted pH to 5 by adding concentrated ammonium hydroxide. The resulting mixture was diluted with 100 mL of water and extracted with EtOAc (30 mL × 3). The combined organic phase was washed with H₂O (20 mL × 3) and brine (20 mL × 3), dried over anhydrous MgSO₄ and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:3) as the mobile phase to provide the desired compounds 16–22.

(4-Methoxyphenyl)(9H-pyrido[3,4-b]indol-1-yl)methanone (16): Yellow solid; yield 35%; mp 182–183 °C; ¹H NMR (600 MHz, CDCl₃) δ 10.46 (brs, 1H), 8.60 (d, J = 5.0 Hz, 1H), 8.45 (d, J = 8.7 Hz, 2H), 8.17 (d, J = 7.8 Hz, 1H), 8.15 (d, J = 5.0 Hz, 1H), 7.62–7.58 (m, 2H), 7.34 (t, J = 7.8 Hz, 1H), 7.03 (d, J = 8.7 Hz, 2H), 3.91 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 193.7, 163.4, 141.1, 138.0, 137.4, 137.1, 134.0, 131.7, 130.4, 129.4, 122.0, 121.0, 120.8, 118.4, 113.6, 112.1, 55.7; HRMS: m/z calcd. for C₁₉H₁₅N₂O₂⁺, 303.1133; found: 303.1130.

(6-Bromo-9H-pyrido[3,4-b]indol-1-yl)(4-methoxyphenyl)methanone (17): Yellow solid; yield 36%; mp 194–195 °C; ¹H NMR (600 MHz, CDCl₃) δ 10.48 (brs, 1H), 8.62 (d, J = 5.0 Hz, 1H), 8.45 (d, J = 8.7 Hz, 2H), 8.29 (d, J = 1.8 Hz, 1H), 8.10 (d, J = 5.0 Hz, 1H), 7.68 (dd, J = 8.7, 1.8 Hz, 1H), 7.48 (d, J = 8.7 Hz, 1H), 7.04 (d, J = 8.7 Hz, 2H), 3.92 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 193.4, 163.5, 139.7, 138.3, 137.6, 137.4, 134.0, 132.1, 130.5, 130.1, 124.7, 122.8, 120.8, 118.5, 113.6, 113.5, 55.7; HRMS: m/z calcd. for C₁₉H₁₄N₂O₂Br⁺, 381.0239; found: 381.0237.

(3-Bromo-4-methoxyphenyl)(9H-pyrido[3,4-b]indol-1-yl)methanone (18): Yellow solid; yield 36%; mp 192–193 °C; ¹H NMR (600 MHz, CDCl₃) δ 10.40 (brs, 1H), 8.69 (d, J = 1.8 Hz, 1H), 8.59 (d, J = 5.0 Hz, 1H), 8.49 (dd, J = 8.7, 2.2 Hz, 1H), 8.15 (m, 2H), 7.60 (m, 2H), 7.35–7.33 (m, 1H), 7.01 (d, J = 8.7 Hz, 1H), 3.99 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 192.2, 159.4, 141.1, 138.1, 137.5, 136.9, 136.5, 133.1, 131.8, 131.4, 129.5, 122.0, 120.9, 118.7, 112.2, 111.5, 110.9, 56.6; HRMS: m/z calcd. for C₁₉H₁₄N₂O₂Br⁺, 381.0239; found: 381.0220.

(3-Bromo-4-methoxyphenyl)(6-bromo-9H-pyrido[3,4-b]indol-1-yl)methanone (19): Yellow solid; yield 31%; mp 195–196 °C; ¹H NMR (600 MHz, CDCl₃) δ 10.50 (brs, 1H), 8.68 (d, J = 1.8 Hz, 1H), 8.59 (d, J = 5.0 Hz, 1H), 8.47 (dd, J = 8.7, 2.2 Hz, 1H), 8.26 (s, 1H), 8.08 (d, J = 5.0 Hz, 1H), 7.66 (dd, J = 8.7, 2.2 Hz, 1H), 7.46 (d, J = 8.7 Hz, 1H), 7.00 (d, J = 8.7 Hz, 1H), 3.99 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 191.9, 159.5, 139.6, 138.3, 137.5, 136.8, 133.2, 132.2, 131.1, 130.7, 124.7, 122.7, 118.7, 113.6, 111.5, 110.9, 56.6; HRMS: m/z calcd. for C₁₉H₁₃N₂O₂Br₂⁺, 458.9344; found: 458.9327.

(3,5-Dibromo-4-methoxyphenyl)(9H-pyrido[3,4-b]indol-1-yl)methanone (20): Yellow solid; yield 38%; mp 197–198 °C; ¹H NMR (600 MHz, CDCl₃) δ 10.38 (brs, 1H), 8.62 (d, J = 5.0 Hz, 1H), 8.59 (s, 1H), 8.20–8.18 (m, 1H), 7.65–7.62 (m, 2H), 7.38–7.36 (m, 1H), 3.98 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 191.5, 157.5, 141.2, 138.4, 137.5, 136.0, 135.6, 132.1, 129.7, 122.1, 121.2, 120.9, 119.2, 118.1, 112.2, 60.9; HRMS: m/z calcd. for C₁₉H₁₃N₂O₂Br₂⁺, 458.9344; found: 458.9333.

(6-Bromo-9H-pyrido[3,4-b]indol-1-yl)(3,5-dibromo-4-methoxyphenyl)methanone (21): Yellow solid; yield 35%; mp 234–235 °C; ¹H NMR (600 MHz, CDCl₃) δ 10.41 (brs, 1H), 8.64 (d, J = 5.0 Hz, 1H), 8.60 (s, 2H), 8.32 (d, J = 1.8 Hz, 1H), 8,16 (d, J = 5.0 Hz, 1H), 7.72 (dd, J = 8.7, 1.8 Hz, 1H), 7.52 (d, J = 8.7 Hz, 1H), 3.99 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 191.3, 157.6, 139.7, 138.7, 137.6, 136.1, 135.4, 134.1, 132.5, 131.0, 124.9, 122.7, 119.3, 118.1, 114.0, 113.8, 60.9; HRMS: m/z calcd. for C₁₉H₁₂N₂O₂Br₃⁺, 536.8449; found: 536.8442.

(7-Bromo-9H-pyrido[3,4-b]indol-1-yl)(3,5-dibromo-4-methoxyphenyl)methanone (22): Yellow solid; yield 36%; mp 216–217 °C; ¹H NMR (600 MHz, CDCl₃) δ 10.38 (brs, 1H), 8.63 (d, J = 5.0 Hz, 1H), 8.59 (s, 2H), 8,16 (d, J = 5.0 Hz, 1H), 8.03 (d, J = 8.2 Hz, 1H), 7.78 (d, J = 1.8 Hz, 1H), 7.48 (dd, J = 8.7, 1.8 Hz, 1H), 3.98 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 191.3, 157.7, 141.9, 138.9, 137.5, 136.1, 135.4, 131.5, 124.7, 123.5, 123.2, 119.9, 119.1, 118.1 115.4, 60.9; HRMS: m/z calcd. for C₁₉H₁₂N₂O₂Br₃⁺, 536.8449; found: 536.8464.

To a solution of the compounds 16–22 (0.5 mmol) in CH₂Cl₂ (10mL) at −78 °C under argon atmosphere, was slowly added dropwise BBr₃ (5 mmol). The reaction mixture was stirred and warmed to r.t. and stirred for 24 h. NaOH solution (5 mL, 2 mol/L) was then slowly added dropwise. After addition, a short period of stirring was continued, and then the solution was acidified with hydrochloric acid (20 mL, 2 mol/L), followed by extraction with CH₂Cl₂ (30 mL × 3). The combined organic phase was washed with H₂O (20 mL × 3) and brine (20 mL × 3), dried over anhydrous MgSO₄ and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography using silica gel as the stationary phase and using ethyl acetate/hexane (1:3) as the mobile phase to provide the desired compounds eudistomins Y₁–Y₇ 23–29.

(4-Hydroxyphenyl)(9H-pyrido[3,4-b]indol-1-yl)methanone (Eudistomins Y₁ 23): Yellow solid; yield 65%; mp 217–218 °C; ¹H NMR (600 MHz, acetone-d₆) δ 11.30(s, 1H), 9.26 (s, 1H), 8.56 (d, J = 5.0 Hz, 1H), 8.47 (d, J = 8.8 Hz, 2H), 8.38 (d, J = 5.0 Hz, 1H), 8.32 (d, J = 7.8 Hz, 1H), 7.88 (d, J = 7.8 Hz, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7. 34 (t, J = 7.8 Hz, 1H), 7.00 (d, J = 8.8 Hz, 2H); ¹³C NMR (151 MHz, acetone-d₆) δ 192.0, 163.1, 142.5, 139.0, 138.4, 137.6, 135.0, 131.0, 130.2, 129.4, 122.7, 122.1, 121.3, 119.7, 115.6, 113.5; HRMS: m/z calcd. for C₁₈H₁₁N₂O₂⁻, 287.0821; found: 287.0830.

(6-Bromo-9H-pyrido[3,4-b]indol-1-yl)(4-hydroxyphenyl)methanone (Eudistomins Y₂ 24): Yellow solid; yield 68%; mp 247–248 °C; ¹H NMR (600 MHz, acetone-d₆) δ 11.46 (s, 1H), 9.30 (s, 1H), 8.60 (d, J = 5.0 Hz, 1H), 8.53 (d, J = 1.8 Hz, 1H), 8.47 (d, J =8.8 Hz, 2H), 8.43 (d, J = 5.0 Hz, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.76 (dd, J = 8.8, 1.8 Hz, 1H), 7.08 (d, J = 8.8 Hz, 2H); ¹³C NMR (151 MHz, acetone-d₆) δ 192.2, 162.8, 141.1, 139.0, 138.5, 137.5, 135.1, 132.3, 131.1, 130.0, 125.4, 123.5, 119.1, 115.5, 113.5, 112.0; HRMS: m/z calcd. for C₁₈H₁₀N₂O₂Br⁻, 364.9926; found: 364.9925.

(3-Bromo-4-hydroxyphenyl)(9H-pyrido[3,4-b]indol-1-yl)methanone (Eudistomins Y₃ 25): Yellow solid; yield 63%; mp 231–232 °C; ¹H NMR (600 MHz, DMSO-d₆) δ 11.99 (s, 1H), 11.28 (s, 1H), 8.57 (d, J = 2.2 Hz, 1H), 8.54 (d, J = 4.4 Hz, 1H), 8.43 (d, J = 4.4 Hz, 1H), 8.32 (d, J = 7.7 Hz, 1H), 8.24 (dd, J = 8.8, 2.2 Hz, 1H), 7.79 (d, J = 8.8 Hz, 1H), 7.60 (t, J = 7.7 Hz, 1H), 7.31 (t, J = 7.7 Hz, 1H), 7.13 (d, J = 7.7 Hz, 1H); ¹³C NMR (151 MHz, DMSO-d₆) δ 190.1, 158.2, 141.6, 137.0, 136.7, 136.4, 135.8, 132.5, 131.0, 129.6, 128.9, 121.8, 120.1, 120.0, 118.7, 115.6, 112.9, 108.8; HRMS: m/z calcd. for C₁₈H₁₀N₂O₂Br⁻, 364.9926; found: 364.9932.

(3-Bromo-4-hydroxyphenyl)(6-bromo-9H-pyrido[3,4-b]indol-1-yl)methanone (Eudistomins Y₄ 26): Yellow solid; yield 66%; mp 265–266 °C; ¹H NMR (600 MHz, DMSO-d₆) δ 12.13 (s, 1H), 11.30 (s, 1H), 8.61 (s, 1H), 8.57 (d, J = 5.5 Hz, 1H), 8.56 (s, 1H), 8.50 (d, J = 5.5 Hz, 1H), 8.23 (dd, J = 8.8, 2.2 Hz, 1H), 7.75–7.72 (m, 1H), 7.13 (d, J = 8.8 Hz, 1H); ¹³C NMR (151 MHz, DMSO-d₆) δ 189.9, 158.3, 140.3, 137.3, 137.1, 136.4, 135.9, 132.5, 131.4, 129.9, 129.4, 124.5, 122.0, 119.1, 115.6, 114.9, 112.2, 108.8; HRMS: m/z calcd. for C₁₈H₉N₂O₂Br₂⁻, 442.9031; found: 442.9012.

(3,5-Dibromo-4-hydroxyphenyl)(9H-pyrido[3,4-b]indol-1-yl)methanone (Eudistomins Y₅ 27): Yellow solid; yield 66%; mp 267–268 °C; ¹H NMR (600 MHz, DMSO-d₆) δ 12.04 (s, 1H), 10.96 (s, 1H), 8.57 (d, J = 5.5 Hz, 1H), 8.54 (s, 2H), 8.47 (d, J = 5.5 Hz, 1H), 8.33 (d, J = 7.3 Hz, 1H), 7.81 (d, J = 8.2 Hz, 1H), 7.61 (t, J = 7.3 Hz, 1H), 7.32 (t, J = 7.3 Hz, 1H); ¹³C NMR (151 MHz, DMSO-d₆) δ 188.9, 154.5, 141.7, 137.1, 136.0, 135.9, 135.4, 131.2, 131.0, 129.0, 121.9, 120.3, 120.0, 119.1, 113.0, 110.9; HRMS: m/z calcd. for C₁₈H₉N₂O₂Br₂⁻, 442.9031; found: 442.9030.

(6-Bromo-9H-pyrido[3,4-b]indol-1-yl)(3,5-dibromo-4-hydroxyphenyl)methanone (Eudistomins Y₆ 28): Yellow solid; yield 62%; mp 277–278 °C; ¹H NMR (600 MHz, DMSO-d₆) δ 12.16 (s, 1H), 10.97 (brs, 1H), 8.61 (s, 1H), 8.59 (d, J = 4.4 Hz, 1H), 8.52 (s, 2H), 8.51 (d, J = 4.4 Hz, 1H), 7.76 (d, J = 8.8 Hz, 1H), 7.73 (dd, J = 8.8, 2.2 Hz, 1H); ¹³C NMR (151 MHz, DMSO-d₆) δ 188.8, 154.6, 140.4, 137.4, 136.4, 136.0, 135.4, 131.5, 130.8, 130.1, 124.5, 122.0, 119.6, 115.0, 112.3, 110.9; HRMS: m/z calcd. for C₁₈H₈N₂O₂Br₃⁻, 520.8136; found: 520.8134.

(7-Bromo-9H-pyrido[3,4-b]indol-1-yl)(3,5-dibromo-4-hydroxyphenyl)methanone (Eudistomins Y₇ 29): Yellow solid; yield 62%; mp 297–298 °C; ¹H NMR (600 MHz, DMSO-d₆) δ 12.11 (s, 1H), 10.96 (brs, 1H), 8.58 (d, J = 4.6 Hz, 1H), 8.53 (s, 2H), 8.46 (d, J = 4.6 Hz, 1H), 8.27 (d, J = 8.2 Hz, 1H), 7.97 (d, J = 1.8 Hz, 1H), 7.45 (dd, J = 8.2, 1.8 Hz, 1H); ¹³C NMR (151 MHz, DMSO-d₆) δ 188.8, 154.6, 142.5, 137.7, 136.3, 136.0, 135.4, 130.8, 130.6, 123.7, 123.2, 121.8, 119.2, 115.6, 110.8; HRMS: m/z calcd. for C₁₈H₈N₂O₂Br₃⁻, 520.8136; found: 520.8141.