13C NMR (150 MHz, CDCl3) 202.4, 167.3, 152.9, 147.2, 134.5, 124.1, 122.0, 110.8, 61.5, TDZD-8 60.7, 55.8, 49.5, 43.8, 24.5, 14.2. Ethyl 5-(2,3-dimethoxyphenyl)-3-hydroxypentanoate (53) To a solution of ketone 52 (1.45 g, 5.17 mmol) in CH3OH (20 mL) at 0 C was added NaBH4 (110 mg, 2.91 mmol), and the reaction mixture was stirred for 1 h at 0 C, then stirred for 3 h at ambient temperature. demonstrated using dynamic bioluminescence imaging in a human prostate tumor xenograft growing in a rat. studies in a mouse model of prostate cancer to evaluate the efficacy of this compound as a VDA, as evidenced by bioluminescence imaging (BLI) (Scheme 6).8,25,57,68,69 Open in a separate window Scheme 6 Synthesis of benzosuberdiene 68 and phosphate salt 69. Table 1. Inhibition of tubulin polymerization, percent inhibition of colchicine binding, and cytotoxicity of the benzosuberene and dihydronaphthalene analogues. prostate tumor xenograft in the thigh. Top) baseline (no prior drug), center) 4 h after 40 mg/kg 69, and bottom) 24 h after 69. B) Corresponding light emission dynamic curve at baseline (blue), 4 h after 69 (red) and 24 h after 69 (green). C) Normalized BLI signal at various times for the rat in Fig 4 receiving 69 sequentially at 10 mg/kg (black), 40 mg /kg (red) and 30 mg/kg CA4P (green), together with the treatment naive rat in Fig 5 A, B receiving 40 mg/kg 69 (orange). Conclusions: These studies have expanded our SAR knowledge regarding the impact of structural modifications to lead benzosuberene and dihydronaphthalene analogues on inhibition of tubulin polymerization and cytotoxicity against human cancer cell lines. Amongst this group of seventeen new molecules [along with the Maderna compound (68), accessed (in this study) through separate synthesis], emerged several promising analogues (compounds 6, 13, 18, 19, 28, 68) that elicited inhibition (IC50) of tubulin assembly (cell free assay) greater than or comparable to that of the lead natural product CA4 and our lead benzosuberene analogues KGP18 and KGP156. These compounds demonstrated potent cytotoxicity (GI50) against SK-OV-3 (ovarian), NCI-H460 (lung), and DU-145 (prostate) cells typically in the low to mid nM range. Preliminary investigation of water-soluble benzosuberene phosphate prodrug salt 69 at 40 mg/kg revealed vascular disruption in a PC3-DAB2IP-human prostate tumor xenograft based on BLI (Figs. 4 and ?and5),5), which was similar to that obtained with CA4P. Experimental Section Chemistry General Materials and Methods Tetrahydrofuran (THF), carbon tetrachloride, dichloromethane, methanol, dimethylformamide (DMF), and acetonitrile were used TDZD-8 in their anhydrous forms. Reactions were performed under nitrogen gas. Thin-layer chromatography (TLC) plates (precoated glass plates with silica gel 60 F254, 0.25 mm thickness) were used to monitor reactions. Purification of intermediates and products Mouse monoclonal to CD3.4AT3 reacts with CD3, a 20-26 kDa molecule, which is expressed on all mature T lymphocytes (approximately 60-80% of normal human peripheral blood lymphocytes), NK-T cells and some thymocytes. CD3 associated with the T-cell receptor a/b or g/d dimer also plays a role in T-cell activation and signal transduction during antigen recognition was carried out with a Biotage Isolera flash purification system using silica gel (200C400 mesh, 60 ?) or RP-18 pre-packed columns or manually in glass columns. Intermediates and products synthesized were characterized on the basis of their 1H NMR (500 or 600 MHz), 13C NMR (125 or 150 MHz) spectroscopic data using a Varian VNMRS 500 MHz or Bruker DPX 600 MHz instrument. Spectra were recorded in CDCl3, D2O, (CD3)2CO, or CD3OD. All chemicals shifts are expressed in ppm (), and peak patterns are reported as broad (br), singlet (s), doublet (d), triplet (t), quartet (q), pentet (p), sextet (sext), septet (sept), double doublet (dd), double double doublet (ddd), and multiplet (m). Purity of the final compounds was further TDZD-8 analyzed at 25 C using an Agilent 1200 HPLC system with a diode-array detector ( = 190?400 nm), a Zorbax XDB-C18 HPLC column (4.6 mm ?~ 150 mm, 5 m), and a Zorbax reliance cartridge guard-column; Method: solvent A, acetonitrile, solvent B, H2O; gradient, 10% A/ 90% B to 100% A/ 0% B over 0 to 40 min; post-time 10 min; flow rate 1.0mL/min; injection volume 20 L; monitored at wavelengths of 210, 230, 254, 280, and 320 nm. Purity of target molecules (with.