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Article

Synthesis of 3-N-Sugar-substituted-2, 4(1H,3H)-quinazolinedionesas Anti-Angiogenesis Agents

State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China
*
Author to whom correspondence should be addressed.
Molecules 2009, 14(7), 2447-2457; https://doi.org/10.3390/molecules14072447
Submission received: 8 May 2009 / Revised: 27 May 2009 / Accepted: 2 June 2009 / Published: 8 July 2009

Abstract

:
A series of novel 3-N-sugar-substituted quinazolinediones were synthesized through the cyclization of the intermediate 2-aminobenzamides using triphosgene as the condensing reagent. Their anti-angiogenesis activities were investigated. The compound 3-(2'-aminoglucosyl)-2,4-(1H,3H)-quinazolinedione, (5d) showed good anti-angiogenesis activity.

Graphical Abstract

1. Introduction

The development of an effective anti-cancer drug is still a major challenge in the field of drug discovery. It was reported that aminopeptidase N (APN) plays a crucial role in the degradation and invasion of extracellular matrices by fibrosarcoma cells [1]. It is also important in the proliferation and the activation of pathogenic T-cells [2]. Several APN inhibitors were prepared to treat inflammatory disease, autoimmune disease, allogenic rejection reactions and allergies. In addition, APN antagonists were found to specifically inhibit angiogenesis in chorioallantoic membranes and in the retina, thus suppressing tumor growth. Therefore, APN was believed to be involved in angiogenesis and can serve as a target for the development of anti-cancer drugs [3,4,5].
The quinazolinediones have inhibitory activities towards some amino peptidases, such as puromycin-sensitive aminopeptidase (PSA) [5] and aminopeptidase N [1]. One representative compound, PAQ-22 (Figure 1), showed potent and specific PSA inhibiting activity with an IC50 of 0.09 μg/mL [5]. The inhibitory mechanism of these compounds was through non-competition, as revealed by the Lineweaver-Burk plot analysis. Structure-activity relationship studies indicated that tautomerism of the imidobenzoylketone group of the cyclic imide moiety of these kinds of inhibitors was important for the inhibitory activity [5].
Figure 1. Structures of PAQ-22 and the desired 3-N-sugar substituted quinazolinedione derivatives.
Figure 1. Structures of PAQ-22 and the desired 3-N-sugar substituted quinazolinedione derivatives.
Molecules 14 02447 g001
It is reported that glucosamine, a type of amino-sugar, possesses immunosuppressive activity and could be beneficial as an immunosuppressive agent [6,7]. Water-soluble conjugates of glucosamine and glucosamine 6-sulfate were reported showing immunomodulatory and anti-angiogenesis properties, These derivatives of glucosamine could function synergistically to prevent scar tissue formation [8,9,10].
To find novel anti-angiogenesis agents, we have synthesized a series of 3-N-sugar-substituted-2,4-(1H,3H)-quinazolinediones containing amino-sugar moieties.

2. Results and Discussion

Generally 3-N-substituted-2,4-(1H,3H)-quinazolinediones could be synthesized through different intermediates, including 2-amino benzamide [11], 2-ureayl benzoic ester [12,13,14], 2-isocyanato benzoate [15,16] and others [17,18]. Considering the known instability of quinazolinediones and the glucosamine moiety under acidic conditions, 2-amino-(N-sugar-substituted) benzamides were chosen as the key intermediates and a mild condensation condition was adopted in the synthetic route. During the synthesis of sugar-N3-substituted quinazolinediones, the unprotected aminosugar was used in the preparations of 2-amino-(N-sugar-substituted) benzamides. The aminosugars were prepared by the reported method [19]. The general synthesis of the key 2-nitro-(N-sugar-substituted) benzamide intermediates was readily performed by the condensation of 2-nitrobenzoic acid and unprotected aminosugars with EDC(DCC)/HOBt [20,21] (Scheme 1). After formation of amides, the sugar hydroxyl groups were fully acetyled by Ac2O/pyridine at room temperature, and then the nitro group was smoothly reduced by powdered Zn in acetic acid/THF. Finally, cyclization with triphosgene in CH2Cl2 (or ClCH2CH2Cl) was performed to produce the target molecules [20,21].
Scheme 1. The synthetic route to N-sugar-substituted quinazolinedione derivatives.
Scheme 1. The synthetic route to N-sugar-substituted quinazolinedione derivatives.
Molecules 14 02447 g003
To prepare the intermediate 2-nitro-(N-sugar-substituted) benzamides from aminosugars, condensations of protected amino-sugars with 2-nitrobenzoyl chloride or with 2-nitrobenzoic acid activated by DCC (or EDC/HOBt) were attempted, but all these efforts failed. It was reported that the unprotected amino-sugars could be used directly to synthesize related amides [20,21], and following this method, the key N-sugar o-nitrobenzamide intermediates were obtained with isolated yields of 40-47%.
The N-sugar-substituted-2,4-(1H,3H)-quinazolinediones 5a-d were obtained from the 2-amino-benzamides by carbonylation cyclization with carbonyldiimidazole (CDI), triphosgene and ethyl chlorocarbonate [20,21]. Several condensation conditions were tested. The results showed in Table 1 indicated that triphosgene had a higher activity than CDI and ethyl chlorocarbonate (Scheme 2). Therefore, triphosgene was used as the condensation reagent. After deacetylation with NaOMe/ MeOH, the target compounds 6a-d were obtained in about 20% total yields.
Table 1. Optimization of the conditions for the preparation of sugar-substituted quinazolinedine derivatives.
Table 1. Optimization of the conditions for the preparation of sugar-substituted quinazolinedine derivatives.
EntryReactantProductReagentSolventReflux Time (h)Isolated Yield (%)
14a5aCDI aTHF48N.R b
24a5aCDI aClCH2CH2Cl48N.R b
34a5atriphosgeneCH2Cl21289%
44a5atriphosgeneClCH2CH2Cl685%
54b5bCDI aTHF48N.R b
64b5bClCO2EtCH3CN48N.R b
74b5btriphosgeneCH2Cl21284%
84b5btriphosgeneClCH2CH2Cl888%
a CDI: carbonyldiimidazole; b N.R: no reaction.
The obtained N-sugar substituted-2,4-(1H,3H)-quinazolinediones 5a-d and 6a-d were primarily assayed for their angiogenesis inhibition activity using the chick chorioallantoic membrane (CAM) model [22]. Only compound 5d showed good inhibitory activity to the neovascularization of chick in vivo (Figure 2).
Figure 2. Inbibition of angiogenesis by compound 5d.
Figure 2. Inbibition of angiogenesis by compound 5d.
Molecules 14 02447 g002

3. Experimental

3.1. Instruments and apparatus

1H- and 13C-NMR spectra were recorded on a Varian VXR 300 MHz spectrometer with Me4Si as the internal standard and CDCl3 or Me2SO-d6 as solvent. Optical rotations were measured at 25 °C with an AA-10R polarimeter. The progress of reactions was monitored by silica-gel GF254 TLC plates. Detection was performed by examination under UV light and by 15% H2SO4 in EtOH. Preparative TLC was performed on silica-gel GF254 plates and column chromatography was on silica-gel H. The inhibitory activity of anti-angiogenesic was assayed under chick chorioallantoic membrances (CAM) model, monitored by biological dissection microscope of DM-1 and recorded with a NIKON S610 digital camera.

3.2. Preparation of 3-N-sugar-substituted-2,4-(1H,3H)-quinazolinediones

3.2.1. Synthesis and Spectral Data of 2a-c

Aminosugar 1a-c (2.9 g, 15 mmol), o-nitrobenzoic acid (3.2 g, 16.5 mmol) and 1-hydroxy benzotriazole (HOBt) (4.9 g, 36.3 mmol) were dissolved in DMF (80 mL). The mixture was cooled to 0 °C and stirred for 30 min. Then a solution of dicyclohexylcarbodiimide (DCC, 3.8 g, 18.2 mmol) in DMF (15 mL) was added dropwise. The mixture was stirred for 18 h at room temperature and filtered. The filtrate was evaporated to dryness under reduced pressure, and the residue was purified by column chromatography on silica gel to give 2a-c.
Methyl-6-(o-nitro)benzamidyl-6-deoxy-α-D-glucopyranoside (2a). White flocculant crystals; Yield: 47.0%; mp: 230-233 °C; [a]D +116o (c 1.01, MeOH); 1H-NMR δ (ppm) (DMSO-d6): 3.18 (m, 1H, H-6b), 3.34 (m, 1H, H-2), 3.38 (s, 3H, OCH3), 3.47 (m, 2H, H-4,5), 3.61 (m, 1H, H-3), 3.85 (m, 1H, H-5), 4.51 (d, 1H, J1, 2 1.2Hz, H-1), 4.67 (d, 1H, J 6.0 Hz, H-OH), 4.79 (d, 1H, J 5.1 Hz, H-OH), 4.94 (d, 1H, J 5.4 Hz, H-OH), 7.55-8.03 (m, 4H, Ph), 8.76 (t, 1H, J5.1Hz, H-NH); 13C-NMR δ (ppm) (DMSO-d6): 40.8 (C-6), 54.4 (OCH3), 70.3 (C-5), 71.9 (C-3), 72.3 (C-2), 73.0 (C-4), 99.7 (C-1), 124.0 (Ph), 129.1 (Ph), 130.6 (Ph), 132.6 (Ph), 133.5 (Ph) , 147.1 (Ph), 165.1 (C=O); ESI-TOF-MS: [M+1]+ m/z 343.0; [M+Na]+ m/z 365.0.
Methyl-6-(o-nitro)benzamidyl-6-deoxy-α-D-galactopyranoside (2b). White flocculant crystals; Yield: 42.3%; mp: 218- 220 °C; [a]D +52o (c 1.01, DMSO); 1H-NMR δ (ppm) (DMSO-d6): 3.28 (s, 3H, OCH3), 3.01-3.77 (m, 5H, sugar-H), 4.57 (m, 3H, sugar-H), 7.57-8.03 (m, 4H, Ph), 8.82 (d, 1H, H-NH); 13C-NMR δ (ppm) (DMSO-d6): 45.5 (C-6), 54.6 (OCH3), 68.3 (C-2), 68.4 (C-5), 69.3 (C-4), 69.4 (C-3), 100.2 (C-1), 124.0 (Ph), 129.0 (Ph), 130.7 (Ph), 132.4 (Ph), 133.5 (Ph), 147.1 (Ph), 165.7 (C=O); ESI-TOF-MS: [M+1]+ m/z 343.0; [M+Na]+ m/z 365.0.
Methyl-6-(o-nitro)benzamidyl-6-deoxy-α-D-mannopyranoside (2c). White flocculant crystals; Yield: 47.0%; mp: 188-189 °C; [a]D +56o (c 1.01, CHCl3); 1H-NMR δ (ppm) (DMSO-d6): 3.13-3.49 (m, 5H, H-sugar), 3.39 (s, 3H, OCH3), 4.60 (m, 1H, H-sugar), 4.50 (s, 1H, H-1), 4.77 (d, J 8.7 Hz, H-OH), 4.78(d, J 4.8 Hz, H-OH), 4.92 (d, J 5.4 Hz, H-OH), 7.54-8.03 (m, 4H, Ph), 8.76 (t, 1H, J 5.1 Hz, H-NH); 13C-NMR δ (ppm) (DMSO-d6): 41.0 (C-6), 54.1 (OCH3), 69.6 (C-4), 70.2 (C-5), 70.6 (C-3), 71.4 (C-2), 101.0 (C-1), 124.0 (Ph), 129.1 (Ph), 130.6 (Ph), 132.6 (Ph), 133.5 (Ph), 147.1 (Ph), 165.9 (C=O); ESI-TOF-MS: [M+1]+ m/z 343.0; [M+Na]+ m/z 365.0.

3.2.2. Synthesis and Spectral Data of 2-(o-nitro)benzamidyl-2-deoxy-β-D-glucopyranose (2d)

Glucosamine hydrochloride (7.8 g, 36 mmol) and sodium methoxide (2.25 g, 41.7 mmol) were added to methanol (100 mL). The mixture was stirred for 20 min and then evaporated to dryness under vacuum. The residue was dissolved in DMF (200 mL), followed by the addition of o-nitrobenzoic acid (5.1 g, 30 mmol) and 1-hydroxybenzotriazole (HOBt, 9.5 g, 72 mmol). The mixture was cooled to 0 °C and stirred for 30 min. Then the solution of dicyclohexylcarbodiimide (DCC, 6.9 g, 36 mmol) in DMF (25 mL) was added dropwise. The mixture was stirred for 20 h at room temperature and filtered. The filtrate was evaporated to dryness under reduced pressure, and the residue was purified through column chromatography on silica gel to yield 5.5 g of white flocculant crystals of 2d; yield: 47%; mp: 208-212 °C; [a]D +40o (c 1.01, MeOH); 1H-NMR δ (ppm) (DMSO-d6): 3.28 (m, 1 H, H-sugar), 3.43-3.83 (m, 5 H, H-sugar), 4.55 (t, 1H, J 5.7 Hz, H-OH), 4.80 (d, 1H, J5.4Hz, H-OH), 5.05 (d, 1H, J 5.4 Hz, H-OH),5.18 (s, 1H, H-1), 6.62 (d, 1H, J 4.2 Hz H-OH), 7.73-8.10 (m, 4H, Ph), 8.64(d, 1H, J 8.1 Hz, H-NH); 13C-NMR δ(ppm) (75MHz, DMSO-d6): 55.1, 61.1, 70.1, 71.1, 72.1 (C of sugar ring), 90.4 (C-1), 123.8 (Ph), 129.5 (Ph), 130.6 (Ph), 132.4 (Ph), 133.1 (Ph), 147.3(Ph), 165.5(C=O); ESI-TOF-MS: [M+1]+ m/z 329.0; [M+Na]+ m/z 351.0.

3.2.3. Synthesis and Spectral Data of 3a-d

The appropriate 2-nitro-(N-sugar-substituted) benzamide 2a-d (2.0 g) was dissolved in pyridine (50 mL), followed by the addition of acetic anhydride (25 mL). The solution was stirred at room temperature overnight and evaporated to dryness under reduced pressure. The residue was dissolved in ethyl acetate and washed sequentially with saturated sodium hydrogen carbonate solution, saturated brine and water. The organic layer was dried over anhydrous Na2SO4 and evaporated under reduced pressure to give 3a-d as yellow solids.
Methyl-6-(o-nitro-)benzamidyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-glucopyranoside (3a). Yield: 95.0 %; mp: 77-79 oC; [a]D +104 o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.02 (s, 3H, Ac), 2.08 (s, 3H, Ac), 2.10 (s, 3H, Ac), 3.42 (s, 3H, OCH3), 3.61 (m, 1H, H-6), 3.79 (m, 1H, H-5), 4.04 (m, 1H, H-5), 4.83 (dd, 1H, J1,2 3.6 Hz, J2,3 10.2 Hz, H-2), 4.91 (d, 1H, J1, 2 3.6 Hz, H-1), 5.02 (t, 1H, J 9.9 Hz, H-3), 5.48 (t, 1H, J 9.9 Hz, H-4), 6.26 (t, 1H, J 5.7 Hz, H-NH), 7.27-8.09 (m, 4H, Ph); 13C-NMR δ (ppm) (CDCl3): 20.7 (CH3CO), 39.4 (C-6), 55.0 (OCH3), 67.6 (C-5), 69.2 (C-3), 69.8 (C-2), 70.9 (C-4), 96.8 (C-1), 124.5 (Ph), 129.0 (Ph), 130.5 (Ph), 132.7 (Ph), 133.9 (Ph), 146.2 (Ph), 166.6 (C=O), 169.9 (CH3CO), 170.2 (CH3CO), 170.3 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 469.1; [M+Na]+ m/z 491.1.
Methyl-6-(o-nitro-)benzamidyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-galactopyranoside (3b). Yield: 98.0%; mp: 87-90 °C; [a]D+68o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 1.99 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.18 (s, 3H, Ac), 3.42(s, 3H, OCH3), 3.54 (m, 2H, H-6), 4.22 (t, 1H, J 6.6 Hz, H-5), 4.98 (d, 1H, J1, 2 3.0 Hz, H-1), 5.16 (dd, 1H, J1,2 3.0 Hz, J2,3 10.8 Hz, H-2), 5.37-5.46 (m, 2H, H-4, H-3), 6.42 (br, 1H, H-NH), 7.38-8.06 (m, 4H, Ph); 13C-NMR δ (ppm) (CDCl3): 20.6 (CH3CO), 20.7 (CH3CO), 20.8 (CH3CO), 39.3 (C-6), 55.7 (OCH3), 66.5 (C-5), 67.4 (C-3), 68.2 (C-2), 69.0 (C-4), 97.2 (C-1), 124.5 (Ph), 128.6 (Ph), 130.6 (Ph), 132.5 (Ph), 133.8 (Ph), 146.4 (Ph), 166.6 (C=O), 169.7 (CH3CO), 170.4 (CH3CO), 170.8 (CH3CO); ESI-TOF-MS: [M+Na]+ m/z 491.0.
Methyl-6-(o-nitro-)benzamidyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-mannopyranoside (3c). Yield: 91.3%; mp:162-164 °C; [a]D+28o (c 1.10, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.00 (s, 3H, Ac), 2.11 (s, 3H, Ac), 2.14 (s, 3H, Ac), 3.38 (s, 3H, OCH3), 3.60 (m, 1H, H-6e), 3.78 (m, 2H, H-6a, 5), 4.67 (s, 1H, H-1), 5.20-5.34 (m, 3H, H-2, 3, 4), 6.40 (t, 1H, J 6.6 Hz, H-NH), 7.58-8.09 (m, 4H, Ph); 13C-NMR δ (ppm) (CDCl3): 20.7(CH3CO), 20.8 (CH3CO), 39.8 (C-6), 55.3 (OCH3), 66.8 (C-4), 68.8 (C-5), 68.9 (C-3), 69.6 (C-2), 98.5 (C-1), 124.6 (Ph), 128.7 (Ph), 130.6 (Ph), 132.9 (Ph), 133.7 (Ph), 146.3(Ph), 166.4(C=O), 169.8 (CH3CO), 169.9 (CH3CO), 170.4 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 469.0; [M+Na]+ m/z 491.0.
2-(o-Nitro)benzamidyl-2-deoxy-1,3,4,6-tetra-O-acetyl-β-D-glucopyranose (3d). Yield: 93.0%; mp: 144-148 °C; [a]D +40o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.06 (s, 3H, Ac), 2.11 (s, 3H, Ac), 2.14 (s, 3H, Ac), 2.17 (s, 3H, Ac), 4.05-4.13 (m, 2H, H-6), 4.30 (dd, 1H, J 3.6 Hz, 12.6 Hz, H-4), 4.67 (m, 1H, H-5), 5.30 (m, 2H, H-2,3), 6.14 (d, 1H, J 8.4 Hz, H-1), 6.40 (d, 1H, J 3.6 Hz, H-NH), 7.27-8.09 (m, 4H, Ph);13C-NMR δ (ppm) (CDCl3): 20.5 (CH3CO), 20.7 (CH3CO), 20.8 (CH3CO), 20.9 (CH3CO), 51.9 (OCH3), 61.4 (C-sugar), 67.4 (C-sugar), 69.8 (C-sugar), 70.0 (C-sugar), 90.4(C-1), 124.6 (Ph), 128.7 (Ph), 130.8 (Ph), 132.0 (Ph), 134.0 (Ph), 145.9 (Ph), 166.5 (C=O), 168.7 (CH3CO), 169.9 (CH3CO), 170.7 (CH3CO), 172.4 (CH3CO); ESI-TOF-MS: [M+Na]+ m/z 519.1.

3.2.4. Synthesis and Spectral Data of 4a-c

The appropriate compound 3a-c (2.0 g, 7.4 mmol) was dissolved in THF (50 mL) and acetic acid (5 mL). Under stirring, zinc power (1.3 g, 20 mmol) was added slowly. The mixture was then refluxed for 2 h, cooled to room temperature, and filtered through a short column of silica gel. The eluent was evaporated to dryness under vacuum. The residue was dissolved in ethyl acetate and washed sequentially with saturated sodium hydrogen carbonate solution, saturated brine and water. The organic layer was dried over anhydrous Na2SO4 and evaporated under reduced pressure to afford the compounds 4a-c as yellow solids.
Methyl-6-(o-amino)benzamidyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-glucopyranoside (4a). Yield: 88.0%; mp: 108-110 °C; [a]D +98o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.01 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.09 (s, 3H, Ac), 3.43 (s, 3H, OCH3), 3.61 (m, 1H, H-6), 3.79 (m, 1H, H-5), 4.04 (m, 1H, H-6), 4.83 (dd, 1H, J1,2 3.6 Hz, J2,3 9.6 Hz, H-2), 4.93 (m, 2H, H-1, 3), 5.50 (t, 1H, J 9.6 Hz, H-3), 5.48 (t, 1H, J 9.9 Hz, H-4), 6.45 (t, 1H, H-NH), 6.45-7.36 (m, 4H, Ph); 13C-NMR δ (ppm) (CDCl3): 20.7 (CH3CO), 22.6 (CH3CO), 38.9 (C-6), 55.4 (OCH3), 67.7 (C-5), 69.7 (C-3), 69.9 (C-2), 70.9 (C-4), 96.6 (C-1), 112.2 (Ph), 114.1 (Ph), 127.4 (Ph), 132.9 (Ph), 148.8 (Ph), 169.7 (C=O), 170.0 (CH3CO), 170.1 (CH3CO), 170.2 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 439.1; [M+Na]+ m/z 461.1.
Methyl-6-(o-amino)benzamidyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-galactopyranoside (4b). Yield: 78.0%; mp: 78-80 °C; [a]D +28o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.00 (s, 3H, Ac), 2.10 (s, 3H, Ac), 2.20 (s, 3H, Ac), 3.39 (s, 3H, OCH3), 3.44 (t, 1H, J 6.6 Hz, H-5), 3.60 (dd, 1H, J5,6e 6.9 Hz, J6a,6e 13.5 Hz, H-6e), 4.15 (dd, 1H, J5,6a 6.9 Hz, J6a, 6e 13.5 Hz, H-6a), 5.00 (d, 1H, J1, 2 3.6 Hz, H-1), 5.18 (dd, 1H, J1,2 3.6 Hz, J2,3 10.8 Hz, H-2), 5.37 (dd, 1H, J3,4 3.3 Hz, J2,3 10.8 Hz, H-3), 5.45 (d, 1H, J 3.3 Hz, H-4), 6.49 (t, 1H, J 6.3 Hz, H-NH), 6.63-7.33 (m, 4H, Ph); 13C-NMR δ (ppm) (CDCl3): 20.6 (CH3CO),20.7 (CH3CO), 20.8 (CH3CO), 38.8 (C-6), 55.0 (OCH3), 66.7 (C-5), 67.5 (C-3), 68.3 (C-2), 69.4 (C-4), 97.2 (C-1), 115.2 (Ph), 116.7 (Ph), 117.4 (Ph), 127.0 (Ph), 132.5 (Ph), 148.9 (Ph), 169.2 (C=O), 169.8 (CH3CO) , 170.5 (CH3CO), 171.0 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 439.1; [M+Na]+ m/z 461.1.
Methyl-6-(o-amino)benzamidyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-mannopyranoside (4c). Yield: 77.6%; mp: 150-154 °C; [a]D +40o (c 1.10, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.00 (s, 3H, Ac), 2.10 (s, 3H, Ac), 2.12 (s, 3H, Ac), 3.37 (s, 3H, OCH3), 3.40 (m, 1H, H-6e), 3.85-3.98 (m, 2H, H-5, 6a), 4.70 (s, 1H, H-1), 5.19-5.25 (m, 2H, H-2, 4), 5.36 (dd, 1H, J3,2 3.3 Hz, J3,4 10.8 Hz, H-3), 6.56 (t, 1H, J 5.4 Hz, H-NH), 6.63-7.37 (m, 4H, Ph); 13C-NMR δ (ppm) (CDCl3): 20.7 (CH3CO), 20.8 (CH3CO), 39.3 (C-6), 55.3 (OCH3), 67.1 (C-4), 68.8 (C-5), 68.9 (C-3), 69.6 (C-2), 98.4 (C-1), 115.8 (Ph), 116.5 (Ph), 117.3 (Ph), 126.9 (Ph), 132.4 (Ph), 148.8 (Ph), 169.2(C=O), 169.9 (CH3CO), 170.0 (CH3CO), 170.3 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 439.1; [M+Na]+ m/z 461.1.

3.2.5. Synthesis and Spectral Data of 4d

Compound 3d (200 mg, 0.4 mmol) was dissolved in methanol (30 mL), and 40% Pd(OH)2 (20 mg) was added. Catalytic hydrogenation was carried out at 4.5 atm of pressure for 6 hours. The solid was filtered and the filtrate was evaporated to dryness to afford 180 mg of 4d, yield: 95.0%; mp: 158-160 °C; 1H-NMR δ (ppm) (CDCl3): 2.05 (s, 3H, Ac), 2.07 (s, 3H, Ac), 2.11 (s, 3H, Ac), 2.18 (s, 3H, Ac), 4.01-4.16 (m, 2H, H-5, 6), 4.30 (dd, 1H, J 3.6 Hz, 12.3Hz ), 4.67 (m, 1H, H-5), 5.26-5.42 (m, 2H), 6.25 (d, 1H, J 8.7 Hz, H-1), 6.31 (d, 1H, J 3.6 Hz, H-NH), 7.27-8.09 (m, 4H, Ph); 13C-NMR δ (ppm) (CDCl3) 20.6 (CH3CO), 20.7 (CH3CO), 20.8 (CH3CO), 20.9 (CH3CO), 51.3 (OCH3), 61.5, 67.4, 69.7, 70.6(C of sugar ring), 90.6 (C-1), 114.3 (Ph), 116.7 (Ph), 117.4 (Ph), 127.0 (Ph), 132.9 (Ph), 149.0 (Ph), 168.7 (C=O), 168.8 (CH3CO), 169.1 (CH3CO), 170.7 (CH3CO), 172.1 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 467.1; [M+Na]+ m/z 489.1.

3.2.6. Synthesis and Spectral Data of 5a-d

Compounds 4a-d (300 mg) were dissolved in ClCH2CH2Cl (50 Ll), then triphosgene (140 mg, 0.54 mmol) was added. The mixture was refluxed for 6h and cooled to room temperature. CH2Cl2 (50 mL) was added and the organic layer was washed with saturated sodium hydrogen carbonate solution, saturated brine and water. The organic layer was dried over anhydrous Na2SO4, evaporated under reduced pressure to dryness, and purified with column chromatography on silica gel to yield white solids of 5a-d.
Methyl-6-(N3-)quinazolinedionyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-glucopyranoside (5a). Yield: 89.0%; mp: 118-120 °C; [a]D +108o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.01 (s, 3H, Ac), 2.05 (s, 3H, Ac), 2.18 (s, 3H, Ac), 3.20 (s, 3H, OCH3), 4.14 (dd, 1H, J6e, 5 3.9Hz, J6a, 6e 13.5 Hz, H-6e), 4.30 (m, 1H, H-5), 4.51 (dd, 1H, J6a, 5 8.4 Hz, J6a, 6e 12.9 Hz, H-6a), 4.90 (d, 1H, J1, 2 3.6 Hz, H-1), 4.94 (dd, 1H, J1,2 8.4 Hz, J2,312.9 Hz, H-2),5.09 (t, 1H, J 9.3 Hz, H-4), 5.48 (t, 1H, J 9.3 Hz, H-3), 7.16-8.16 (m, 4H, Ph). 10.2 (s, 1H, H-NH); 13C-NMR δ (ppm) (CDCl3): 20.7 (CH3CO), 39.4 (C-6), 55.0 (OCH3), 67.6 (C-5), 69.2 (C-3), 69.8 (C-2), 70.9 (C-4), 96.8 (C-1), 124.5 (Ph), 129.0 (Ph), 30.5 (Ph), 132.7 (Ph), 133.9 (Ph), 146.2 (Ph), 166.6 (C=O), 169.9 (CH3CO), 170.2 (CH3CO), 170.3 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 465.0; [M+Na]+ m/z 487.0.
Methyl-6-(N3-)quinazolinedionyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-galactopyranoside (5b). Yield: 89.0%, mp: 216-219 °C; [a]D +220o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 1.96 (s, 3H, Ac), 2.08 (s, 3H, Ac), 2.26 (s, 3H, Ac), 3.34 (s, 3H, OCH3), 4.29 (m, 2H, H-5,6e), 4.52 (t, 1H, J 6.6 Hz, H-6a), 5.00 (d, 1H, J1, 2 3.3 Hz, H-1), 5.20 (dd, 1H, J1,2 3.3 Hz, J2,3 10.8 Hz, H-2), 5.33 (dd, 1H, J3,4 3.3 Hz, J2,3 10.8 Hz, H-3), 5.38 (d, 1H, J 3.0 Hz, H-4), 7.10-8.13 (m, 4H, Ph), 10.1 (s, 1H, H-NH); 13C-NMR δ (ppm) (CDCl3): 20.7 (s, 3H, Ac), 20.8 (s, 3H, Ac), 20.9 (CH3CO), 39.8 (C-6), 55.2 (OCH3), 65.2 (C-5), 67.9 (C-3), 68.1 (C-2), 68.2 (C-4), 97.0 (C-1), 114.2 (Ph), 115.0 (Ph), 123.6 (Ph), 128.5 (Ph), 135.3 (Ph), 138.4 (Ph), 151.6 (C=O), 162.2 (C=O), 170.1 (CH3CO), 170.4 (CH3CO), 170.8 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 465.1; [M+Na]+ m/z 487.1.
Methyl-6-(N3-)quinazolinedionyl-6-deoxy-2,3,4-tri-O-acetyl-α-D-mannopyranoside (5c). Yield: 81.8%; mp: 78-82 °C; [a]D +32o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.01 (s, 3H, Ac), 2.07, (s, 3H, Ac) 2.16 (s, 3H, Ac), 3.19 (s, 3H, OCH3), 4.14 (dd, 1H, J6e, 5 3.9 Hz, J6a, 6e 13.5 Hz, H-6e), 4.30 (m, 1H, H-5), 4.51 (dd, 1H, J6a, 5 8.4 Hz, J6a, 6e 13.5 Hz, H-6a), 4.60 (dd, 1H, J2,3 2.1 Hz, J1,2 4.8 Hz, H-2), 5.21 (d, 1H, J1, 2 2.1 Hz, H-1), 5.32-8.34 (m, 2H, H-4,3), 7.21-8.16 (m, 4H, Ph). 10.7 (s, 1H, H-NH); 13C-NMR δ (ppm) (CDCl3): 20.6 (CH3CO), 20.7 (CH3CO), 20.9 (CH3CO), 41.9 (C-6), 54.8 (OCH3), 67.3 (C-5), 69.0 (C-3), 69.1 (C-2), 69.5 (C-4), 98.1 (C-1), 114.1 (Ph), 115.2 (Ph), 123.5 (Ph), 128.4 (Ph), 135.2 (Ph), 138.5 (Ph), 152.0 (C=O), 162.2 (C=O), 169.9 (CH3CO), 170.1 (CH3CO), 170.2 (CH3CO); ESI-TOF-MS: [M+1]+ m/z 465.0; [M+Na]+ m/z 487.0.
2-(N3-)quinazolinedionyl-2-deoxy-1,3,4,6-tetra-O-acetyl-β-D-glucopyranoside (5d). Yield: 57.1%; mp: 193-195 °C; [a]D +88o (c 1.01, CHCl3); 1H-NMR δ (ppm) (CDCl3): 2.04 (s, 3H, Ac), 2.05 (s, 3H, Ac), 2.11 (s, 3H, Ac), 2.18 (s, 3H, Ac), 4.02-4.32 (m, 4H, H-sugar), 4.67 (m, 1H, H-5), 5.26-5.42 (m, 2H, H-sugar), 6.33 (d, 1H, J 3.6 Hz, H-sugar), 6.37 (d, 1H, J 8.1 Hz, H-1), 7.01-8.39 (m, 4H, Ph), 10.2 (ds, 1H, H-NH); 13C-NMR δ (ppm) (CDCl3): 20.6 (CH3CO), 20.7 (CH3CO), 20.8 (CH3CO), 20.9 (CH3CO), 51.8 (OCH3), 61.2, 61.5, 67.2, 69.7, 70.6 (C of sugar ring), 90.4 (C-1), 118.3 (Ph), 121.9 (Ph), 126.3 (Ph), 133.3 (Ph), 140.4 (Ph), 153.8 (Ph), 168.6 (C=O), 169.1 (CH3CO), 172.1 (CH3CO), 179.9 (C=O); ESI-TOF-MS: [M+NH4]+ m/z 510.0; [M+Na]+ m/z 515.0; [M+K]+ m/z 530.9.

3.2.7. Synthesis and Spectral Data of 6a-d

The appropriate intermediate 5a-d (130 mg) was dissolved in MeOH(20 mL) and sodium methoxide (10 mg, 0.18 mmol) was added and the mixture stirred for 30 min. The solution was then neutralized to pH 6-7 by with resin and filtered. The filtrate was evaporated to dryness to obtain light yellow solid of 6a-d.
Methyl-6-(N3-)quinazolinedionyl-6-deoxy-α-D-glucopyranoside (6a). Yield: 98%; mp: 137-142 °C; [a]D +60o (c 1.01, DMSO); 1H-NMR δ (ppm) (DMSO-d6): 2.97 (s, 3H, OCH3), 3.03 (m, 1H), 3.17 (m, 1H), 3.35 (m, 1H), 3.82 (m, 1H), 4.05 (m, 1H), 4.19 (m, 1H), 4.40 (d, 1H, J1, 2 3.0 Hz, H-1); 13C-NMR δ (ppm) (DMSO-d6): 41.8 (C-6), 53.7 (OCH3), 67.2 (C-5), 71.9 (C-3), 73.2 (C-2), 73.8 (C-4), 99.5 (C-1), 113.8 (Ph), 115.8 (Ph), 122.0 (Ph), 27.3 (Ph), 134.7 (Ph), 140.7(Ph), 150.9 (C=O), 162.3 (C=O); ESI-TOF-MS: [M+1]+ m/z 339.0; [M+Na]+ m/z 361.0.
Methyl-6-(N3-)quinazolinedionyl-6-deoxy-α-D-galactopyranoside (6b). Yield: 81.5%; mp: 235-237 °C; [a]D +220o (c 1.01, MeOH); 1H-NMR δ (ppm) (DMSO-d6): 3.34 (s, 3H, OCH3), 3.49-3.58 (m, 2H), 3.65-3.96 (m, 3H), 4.07 (dd, 1H, 1H, J2,3 3.6 Hz, J3,4 9.3 Hz, H-3), , 4.37 (s, 1H, H-1), 4.42-4.59 (m, 3H); 13C-NMR δ (ppm) (DMSO-d6): 40.9 (C-6), 55.2 (OCH3), 67.1, 68.2, 69.0, 70.2 (C of sugar ring), 99.9 (C-1), 104.2 (C-1), 113.7 (Ph), 115.1 (Ph), 122.4 (Ph), 127.4 (Ph), 135.0 (Ph), 139.4(Ph), 150.4 (C=O), 162.2 (C=O); ESI-TOF-MS: [M+1]+ m/z 339.1; [M+Na]+ m/z 361.0.
Methyl-6-(N3-)quinazolinedionyl-6-deoxy-α-D-mannopyranoside (6c). Yield: 81.0%; mp: 169-172oC; [a]D +60o (c 1.01, DMSO); 1H-NMR δ (ppm) (DMSO-d6): 2.93 (s, 3H, OCH3), 3.39-3.54 (m, 3H, H-5, 6a, 6e), 3.76 (m, 1H), 4.06 (dd, 1H, 1H, J2,3 3.6 Hz, J3,4 9.3 Hz, H-3), 4.27 (dd, 1H, J4,5 9.6 Hz, J3,4 13.2 Hz, H-4), 4.37 (s, 1H, H-1); 13C-NMR δ (ppm) (DMSO-d6): 41.8 (C-6), 53.4 (OCH3), 68.2, 70.1, 70.9 (C of sugar ring), 100.8 (C-1), 113.7 (Ph), 115.1 (Ph), 122.4 (Ph), 127.4 (Ph), 134.9 (Ph), 139.5 (Ph), 150.3 (C=O), 162.1 (C=O); ESI-TOF-MS: [M+1]+ m/z 339.0; [M+Na]+ m/z 361.0.
2-(N3-)quinazolinedionyl-2-deoxy-D-glucopyranoside (6d). Yield: 90.9%; mp:174-179 °C; [a]D +20o (c 1.10, DMSO); 1H-NMR δ (ppm) (DMSO-d6): 3.69-3.78 (m, 2H, H-5, H-6), 4.01 (dd, 1H, J6a, 5 3.0 Hz, J6a, 6e 5.4 Hz, H-6a), 4.60 (t, 1H, J 4.8 Hz), 4.78 (dd, 1H, J2,3 2.4 Hz, J2,1 5.1 Hz, H-2), 5.08 (dd, 1H, J2,3 2.4 Hz, J3,41 5.1 Hz, H-3), 5.54 (d, 1H, J2,1 5.1 Hz, H-1), 6.0 (s, 1H, H-OH), 7.18-7.95 (m, 4H, Ph); 13C-NMR δ (ppm) (DMSO-d6): 65.9, 69.3, 70.9, 80.6, 84.6 (C-sugar), 100.8 (C-1), 113.7 (Ph), 115.1 (Ph), 122.7 (Ph), 127.5 (Ph), 135.2 (Ph), 139.5 (Ph), 150.0 (C=O), 162.1 (C=O); ESI-TOF-MS: [M+NH4]+ m/z 338.0; [M+Na]+ m/z 347.0.

3.3. Anti-angiogenesic Inhibitory Activity of the Target Compounds

The eggs were cut and chicken embryos were incubated under 37.5 °C for 7 days. When the CAM‘s diameter had grown to 1-3cm, solutions of the compounds was added to each chicken embryo with PBS as control. The results were recorded by camera under a dissection microscope [22].

4. Conclusions

In summary, several novel 3-N-sugar-substituted quinazolinediones were synthesized and their anti-angiogenesis activities were tested. An efficient method, using triphosgene as the carbonylation condensation reagent, was developed for the synthesis of N-sugar-substituted quinazolinediones. This method might be useful in the future for the preparation of similar derivatives.

Acknowledgements

This work was supported by the National Science Foundation of China (30672525, 20732001).

References and Notes

  1. Fujii, H.; Nakajima, M.; Aoyagi, T. Inhibition of tumor cell invasion and matrix degradation by aminopeptidase inhibitors. Biol. Pharm. Bull. 1996, 19, 6–10. [Google Scholar] [CrossRef]
  2. Siegfried, A.; Janine, T.; Uwe, L. Use of alanyl-aminopeptidase inhibitors and pharmaceutical compositions containing said inhibitors. WO 2004004750.
  3. Renata, P.; Erkki, K.; Renate, K. Aminopeptidase N is a receptor for tumor-homing peptides and a target for inhibiting angiogenesis. Cancer Res. 2000, 60, 722–727. [Google Scholar]
  4. Ikeda, N.; Nakajima, Y.; Tokuhara, T. Clinical significance of amino peptidase N/CD13 expression in human pancreatic carcinoma. Clin. Cancer Res. 2003, 9, 1503–1508. [Google Scholar]
  5. Hashimoto, Y. Structural development of biological response modifiers based on thalidomide. Bioorg. Med. Chem. 2002, 10, 461–479. [Google Scholar] [CrossRef]
  6. Ma, L.; Rudert, W.A.; Harnaha, J.; Wright, M.; Machen, J. Immunosuppressive effects of glucosamine. J. Biol. Chem. 2002, 277, 39343–39349. [Google Scholar]
  7. Shaunak, S.; Thomas, S.; Gianasi, E. Polyvalent dendrimer glucosamine conjugates prevent scar tissue formation. Nat. Biotechnol. 2004, 22, 977–984. [Google Scholar] [CrossRef]
  8. Michael, J.; Gueguen, G.; Vercauteren, J.; Moreau, S. Triplex stability of oligodeox ynucleotides containing substituted quinazoline-2,4-(1H, 3H)-dione. Tetrahedron 1997, 53, 8457–8478. [Google Scholar] [CrossRef]
  9. Park, H.Y.; Choo, M.K; Lee, S.K.; Kim, S.W. Solid-phase combinatorial synthesis and cytotoxicity of 3-aryl-2, 4-quinazolindiones. Bioorg. Med. Chem. 2002, 10, 517–523. [Google Scholar] [CrossRef]
  10. Ryu, C.K.; Shin, K.H.; Seo, J.H.; Kim, H.J. 6-Arylamino-5, 8-quinazolinediones as potent inhibitors of endothelium–depend ent vasorelaxation. Eur. J. Med. Chem. 2002, 37, 77–82. [Google Scholar] [CrossRef]
  11. Kakuta, H.; Koiso, Y.; Takahashi, H.; Nagasawa, K.; Hashimoto, Y. Novel Specific puromycin-sensitive aminopeptidase inhibitors: 3-(2, 6-Diethylphenyl)-2, 4(1H, 3H) quinazolinedione and N-(2, 6-Diethylphenyl)-2-amino-4H-3, 1-benzoxazin-4-one. Heterocycles 2001, 55, 1433–1437. [Google Scholar] [CrossRef]
  12. Canonne, P.; Akssira, M.; Dahdouh, A.; Kasmi, H.; Boumzebra, M. Synthesis of chiral 3-Substituted 2, 4(1H, 3H)-quinazolinediones. Heterocycles 1993, 36, 1305–1314. [Google Scholar] [CrossRef]
  13. Aziane, D.; Soukri, M.; Hakmaoui, A.E. A Convenient Synthesis of 5-and 8-Nitroquinazoline-2, 4-dione derivatives. J. Heterocycl. Chem. 2002, 39, 271–276. [Google Scholar] [CrossRef]
  14. Larksarp, C.; Alper, H. Palladium-catalyzed cyclocarbonylation of o-iodoanilines with hetero-cumulenes: regioselective preparation of 4(3H)-quinazolinone derivatives. J. Org. Chem. 2000, 65, 2773–2777. [Google Scholar] [CrossRef]
  15. Azizian, J.; Mehrdad, M.; Jadidi, K.; Sarrafi, Y. Rearrangement of 4-imino-(1H, 4H)-3,1-benzoxazine-2-ones to 2,4-quinazolinediones via an isocyanate carboxamide intermediate. Tetrahedron Lett. 2000, 41, 5265–5268. [Google Scholar] [CrossRef]
  16. Willis, M.C.; Snell, R.H.; Fletcher, A; Woodward, R.L. Tandem Palladium-Catalyzed Urea Arylation−Intramolecular Ester Amidation: Regioselective Synthesis of 3-Alkylated 2,4-Quinazo linediones. Org. Lett. 2006, 8, 5089–5092. [Google Scholar] [CrossRef]
  17. Reina, J.J.; Maldonado, O.S.; Tabarani, G.; Fieschi, F.; Rojo, J. Mannose glycoconju gates functionalized at positions 1 and 6. binding analysis to DC-SIGN using biosensors. Bioconjug. Chem. 2007, 18, 963–969. [Google Scholar] [CrossRef]
  18. Cottaz, S.; Brimacombe, J.S.; Ferguson, M.A.J. An imino-linked carba-disac charide α--mannosidase inhibitor. Carbohydr. Res. 1993, 247, 341–345. [Google Scholar] [CrossRef]
  19. Shibata, Y.; Kosuge, Y.; Mizukoshi, T.; Ogawa, S. Chemical modification of the sugar part of methyl acarviosin: synthesis and inhibitory activities of nine analogues. Carbohydr. Res. 1992, 228, 377–398. [Google Scholar] [CrossRef]
  20. El-Barbary, A.A.; El-Brollosy, N.R.; Pedersen, E.B. Synthesis of 3'-Azido- and 3'-Amino-2', 3'-dideoxynucleosides from 2, 4-Quinazolinediones. Heterocycles 1994, 38, 2191–2198. [Google Scholar] [CrossRef]
  21. Gouilleux, L.; Fehrentz, J.A.; Winternitz, F.; Martinez, J. Solid phase synthesis of chiral 3-substituted quinazoline-2, 4-diones. Tetrahedron Lett. 1996, 37, 7031–7034. [Google Scholar] [CrossRef]
  22. Yang, X.R.; Zhao, B.Z.; Cui, J.R. Anti-angiogenesis of sodium aescinate in vivo. Zhongguo Xinyao Zazhi 2006, 15, 868–870. [Google Scholar]
  • Sample Availability: Samples of the compounds 6a-d are available from the authors.

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MDPI and ACS Style

Huang, C.; Meng, X.; Cui, J.; Li, Z. Synthesis of 3-N-Sugar-substituted-2, 4(1H,3H)-quinazolinedionesas Anti-Angiogenesis Agents. Molecules 2009, 14, 2447-2457. https://doi.org/10.3390/molecules14072447

AMA Style

Huang C, Meng X, Cui J, Li Z. Synthesis of 3-N-Sugar-substituted-2, 4(1H,3H)-quinazolinedionesas Anti-Angiogenesis Agents. Molecules. 2009; 14(7):2447-2457. https://doi.org/10.3390/molecules14072447

Chicago/Turabian Style

Huang, Conghai, Xiangbao Meng, Jingrong Cui, and Zhongjun Li. 2009. "Synthesis of 3-N-Sugar-substituted-2, 4(1H,3H)-quinazolinedionesas Anti-Angiogenesis Agents" Molecules 14, no. 7: 2447-2457. https://doi.org/10.3390/molecules14072447

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