Journal of Medicinal Chemistry pubs.acs.org/jmc Article 13 (CO-Coum), 156.8 (NHCO ), 152.6, 138.7 (Cq-Ar), 129.7 (2 × (2m, 2H, H-β), 1.46−1.34 (m, 4H, H-γ,H-δ); CNMR(126MHz, 2 CH-Ar), 129.1 (CH-Ar), 128.9 (2 × CH-Ar), 127.9, 114.4 (CH-Ar), (CD ) SO): δ 173.0 (CO H), 165.3 (CONH-Ar), 161.4 (CO-Ar) 3 2 2 113.2 (CH-Ar, Cq-Ar), 103.8 (CH-Ar), 67.3 (OCH -Ph), 66.9 (CH - 156.0 (NHCO ), 146.6 (Cq-Ar), 139.2 (Cq-Ar), 137.3 (Cq-Ar), 2 2 2 Coum), 54.0 (C-α), 41.6 (C-ε), 32.2 (C-β), 30.6 (C-δ), 24.3 (C-γ); 130.6 (CH-Ar), 128.3 (2 × CH-Ar), 127.6 (3 × CH-Ar), 126.2 (Cq- HPLC (method A-2): tR = 13.2 min; HPLC-MS (method B): tR = Ar), 121.9 (CH-Ar), 119.5 (CH-Ar), 116.4 (CH-Ar), 115.5 (CH-Ar), + + 11.8 min; MS: (ESI ), m/z (%): [M + H] = 483.1 (100%), [M − 65.1 (OCH -Ph), 53.6 (C-α), 40.2 (C-ε), 31.1 (C-β), 29.2 (C-δ), + − − 2 CO +H] =439.4(20%);MS:(ESI ),m/z(%):[M−H] =481.3 23.0 (C-γ); HPLC (method A-2): t = 12.8 min; HPLC-MS (method 2 − − R (100%), [M − C H O − H] = 373.2 (50%); HRMS (ESI ) calcd + + 7 8 − max B): tR = 11.8 min; MS: (ESI ), m/z (%): [M + H] = 452.3 (100%), for [C H N O − H] : 481.1616; found: 481.1619; λ = 324 nm, + − − max 25 26 2 8 −1 −1 abs [M−CO2+H]=408.3(20%);MS:(ESI ),m/z(%):[M−H] = λ = 399 nm, ε (λ ) = 9880 M cm , Φ = 0.0904. 450.6 (20%); [2M − H]− = 902.0 (100%); HRMS (ESI−) calcd for em 6 max 2 N -((Benzyloxy)carbonyl-N -(2-(7-(dimethylamino)-2-oxo-2H- − max [C H NO − H] : 450.1670; found: 450.1669; λ = 332 nm, 24 25 3 6 abs chromen-4-yl)acetyl)-L-Lysine (37). From 7-dimethylamino-4-car- λmax= 434 nm, ε (λ ) = 16,700 M−1 cm−1, Φ = 0.0101. boxymethyl-coumarin 29 (0.125 g, 0.5 mmol, 1 equiv), HCl· em 6 max 2 N -((Benzyloxy)carbonyl-N -(3-(3,6-dihydroxy-9H-xanthen-9- Lys(Cbz)-OMe (0.165 g, 0.6 mmol, 1.1 equiv), HBTU (0.220 g, ylidene)propanoyl)-L-lysine (40). From compound 32 (0.384 g, 1.0 0.6 mmol, 1.1 equiv), and DIEA (346 μL, 2.0 mmol, 4 equiv), mmol, 1 equiv), HBTU (0.417 g, 1.1 mmol, 1.1 equiv), and HOBt compound37(0.128g) was obtained by filtration as a yellow powder (0.148 g, 1.1 mmol, 1.1 equiv) were diluted with anhydrous DMF (3 1 in a 50% yield according to general procedure B. H NMR (500 MHz, mL). DIEA (380 μL, 2.2 mmol, 2.2 equiv) was added dropwise, and (CD ) SO): δ 8.55 (d, J = 7.0 Hz, 1H, NH), 7.56 (d, J = 9.0 Hz, 1H, 3 2 this solution was stirred for a few minutes then a solution of HCl· H-5), 7.30 (m, 6H, H-Ph, NH), 6.66 (d, J = 9.0 Hz, 1H, H-6), 6.54 (s, Lys(Cbz)-OMe (0.331 g, 1.0 mmol, 1 equiv) in DMF (1 mL) was 1H, H-8), 6.04 (s, 1H, H-3), 5.00 (s, 2H, CH -Ph), 4.16 (m, 1H, H- 2 added. The solution was stirred overnight at room temperature then α), 3.67 (s, 2H, CH -Coum), 3.00 (m, 8H, H-ε,CH), 1.71, 1.61 2 13 3 the solvent was evaporated, and the residue was diluted in EtOAc (50 (2m, 2H, H-β), 1.39−1.30 (m, 4H, H-γ,H-δ); CNMR(126MHz, mL). The organic layer was washed with brine (3 × 40 mL), dried (CD ) SO): δ 173.4 (CO H), 168.0 (CONH) 160.7, 156.0 (CO- 3 2 2 (Na SO ), filtered, and evaporated. The mixture was diluted with a Coum), 155.3 (NHCO ), 152.8 (Cq-Ar), 151.3 (Cq-N(Me) ), 137.7 2 4 2 2 solution of aqueous LiOH (0.5 M)/THF (40 mL, 1/1) and stirred 2 (Cq-Ar), 128.3 (2 × CH-Ar), 127.7 (3 × CH-Ar), 126.0 (CH-Ar), h at room temperature. After evaporation of THF, the pH was 109.3 (CH-Ar), 108.9 (CH-Ar), 108.2 (Cq-Ar), 97.5 (CH-Ar), 65.1 adjusted to 2 with aqueous HCl solution (1 M) and extracted with (OCH-Ph), 52.00 (C-α), 40.1 (C-ε, 2xNCH ), 38.4 (CH -Coum), 2 3 2 EtOAc (2 × 40 mL). The organic layer was dried (Na SO ), filtered, 30.6 (C-β), 29.0 (C-δ), 22.7 (C-γ); HPLC (method A-2): t = 14.4 2 4 + R and evaporated. The residue was purified by column chromatography min; HPLC-MS (method B): tR = 13.7 min; MS: (ESI ), m/z (%): on silica gel (CH Cl /MeOH/AcOH: 85/15/1). Compound 40 + + 2 2 [M + H] = 510.1 (100%), [M − CO +H] = 466.3 (20%); MS: − − 2 − (0.045 g) was obtained as a red powder in an 8% yield. mp: 106−107 (ESI ), m/z (%): [M − H] = 508.3 (100%): HRMS (ESI ) calcd °C; 1H NMR (126 MHz, (CD ) SO): δ 7.51 (d, J = 8.5 Hz, 1H, H- − max 3 2 for [C H N O − H] : 508.2089; found: 508.2089; λ = 373 nm, 27 31 3 7 abs Ar), 7.41 (d, J = 8.5 Hz, 1H, H-Ar), 7.15 (m, 5H, H-Ph), 6.59−6.45 max −1 −1 λ = 448 nm, Φ = 0.714, ε (λ ) = 16,800 M cm . em max (m, 4H, H-Ar), 5.78 (t, J = 7.0 Hz, 1H, CCH), 4.69 (s, 2H, CH - N6-((Benzyloxy)carbonyl-N2-(2-(7-(diethylamino)-2-oxo-2H- 2 Ph), 4.25 (m, 1H, H-α), 3.31 (d, J = 7.0 Hz, 2H, CH -CH), 2.88 (m, chromen-4-yl)acetyl)-L-lysine (38). From 7-diethylamino-4-carbox- 2 13 ymethyl-coumarin 30 (0.275 g, 1.0 mmol, 1 equiv), HCl·Lys(Cbz)- 2H, H-ε), 1.70, 1.55 (m, 2H, H-β), 1.29−1.18 (m, 4H, H-γ,H-δ); C NMR (126 MHz, (CD ) SO): δ 172.9 (CO H), 164.8 (CONH), OMe(0.360 g, 1.2 mmol, 1.2 equiv), HBTU (0.440 g, 1.2 mmol, 1.2 3 2 2 equiv), and DIEA (865 μL, 5.0 mmol, 5 equiv), compound 38 (0.128 158.2, 157.9 (Cq-3, Cq-6), 156.0 (NHCO), 153.0 (Cq-Ar), 151.3 g) was obtained by filtration as a yellow powder in a 19% yield (Cq-Ar), 138.7 (Cq-Ar), 128.3 (CH-Ar), 128.1 (2 × CH-Ar), 127.2 according to general procedure B. 1H NMR (500 MHz, (CD ) SO): (3 × CH-Ar), 125.7 (Cq-Ar), 123.3, 115.7 (CH-Ar), 114.0, 113.1 3 2 (Cq-Ar), 112.0 (CH-Ar), 110.8 (CH), 103.0 (CH-Ar), 102.4 (CH- δ 8.54 (d, J = 7.5 Hz, 1H, NH), 7.52 (d, J = 9.0 Hz, 1H, H-5), 7.30 Ar), 66.1 (OCH Ph), 52.6 (C-α), 40.6 (C-ε), 36.3 (CH CO), 31.0 (m, 6H, H-Ph, NH), 6.66 (dd, J = 9.0 Hz, J = 2.5 Hz, 1H, H-6), 6.54 2 2 (d, J = 2.5 Hz, 1H, H-8), 5.99 (s, 1H, H-3), 5.00 (s, 2H, CH2-Ph), (C-β), 29.1 (C-δ), 22.6 (C-γ); HPLC (method A-2): tR = 13.7 and 15.1 min; HPLC-MS (method B): t = 12.1 and 12.9 min (2 4.15 (m, 1H, H-α), 3.65 (s, 2H, CH -Coum), 3.41 (q, J = 7.0 Hz, 4H, R 2 + + N(CH CH ) ), 2.96 (m, 2H, H-ε), 1.72, 1.60 (2m, 2H, H-β), 1.45− tautomers); MS: (ESI ), m/z (%): [M + H] = 547.2 (100%); MS: 2 3 2 13 − − − (ESI ), m/z (%): [M − H] = 545.3 (100%), [M − C H O − H] = 1.26 (m, 4H, H-γ,H-δ), 1.10 (t, J = 7.0 Hz, 6H, N(CH CH ) ); C 7 7 2 3 2 − − 438.2 (30%); HRMS (ESI ) calcd for [C H N O − H] : NMR (126 MHz, (CD ) SO): δ 173.4 (CO H), 168.0 (CONH), 30 30 2 8 3 2 2 max max 545.1929; found: 545.1956; λ = 455 nm, λ = 516 nm, ε (λ ) 160.7, 156.0 (CO-Coum), 155.7 (NHCO2), 151.2 (Cq-Ar), 150.3 −1 −1 abs em max (Cq-N(Me) ), 137.3 (Cq-Ar), 128.3 (2 × CH-Ar), 127.7 (3 × CH- = 5140 M cm , Φ = 0.243. 2 N6-((Benzyloxy)carbonyl-N2-(2-(11-oxo-2,3,6,7-tetrahydro- Ar), 126.3 (CH-Ar), 108.8 (CH-Ar), 108.5 (CH-Ar), 107.7 (Cq-Ar), 1H,5H,11H-pyrano[2,3-f]pyrido[3,2,1-ij]quinolin-9-yl))acetyl)-L-ly- 96.8 (CH-Ar), 65.1 (OCH -Ph), 52.0 (C-α), 44.0 (2 × NCH CH ), 2 2 3 sine (41). From compound 31 (0.150 g, 0.50 mmol, 1 equiv), HCl· 40.1 (C-ε), 38.4 (CH -Coum), 30.6 (C-β), 29.0 (C-δ), 22.7 (C-γ), 2 Lys(Cbz)-OMe (0.182 g, 0.55 mmol, 1.1 equiv), HBTU (0.209 g, 12.3 (2 × NCH CH ); HPLC (method A-2): t = 15.9 min; HPLC- 2 3 + R + 0.55 mmol, 1.1 equiv), and DIEA (346 μL, 2.0 mmol, 4 equiv), MS(method B): tR = 14.7 min; MS: (ESI ), m/z (%): [M + H] = + + compound 41 (0.033 g) was obtained by filtration as a yellow powder 538.1 (100%), [2M + H] = 1074.9 (90%), [M − CO +H] = 494.4 − 2 − in a 12% yield according to general procedure B. 1H NMR (500 MHz, (20%); HRMS (ESI ) calcd for [C H N O − H] : 536.2402; max max 29 35 3 7 −1 CDOD): δ 7.40 (m, 5H, H-Ph) 7.15 (s, 1H, H-8), 6.01 (s, 1H, H- found: 536.2402; λ =379nm,λ =446nm,ε(λ )=18,600M 3 −1 abs em max 10), 5.06 (s, 2H, CH -Ph), 4.40 (m, 1H, H-α), 3.72 (s, 2H, CH - cm , Φ = 0.749. 2 2 6 2 Coum), 3.27 (m, 4H, CH N), 2.84−2.71 (m, 6H, 2 × CH -Ar, H-ε), N -((Benzyloxy)carbonyl-N -(2-oxo-1,2-dihydroquinoline-4-car- 2 2 bonyl)-L-lysine (39). From 4-carboxy-quinolin-2-one (0.189 g, 1.0 1.98−1.91 (m, 5H, 2 × CH -CH -CH ,H-β), 1.75 (m, 1H, H-β), 2 132 2 mmol, 1 equiv), HCl·Lys(Cbz)-OMe (0.364 g, 1.1 mmol, 1.1 equiv), 1.62−1.41 (m, 4H, H-γ,H-δ); C NMR (126 MHz, CD OD): δ 3 HBTU(0.420g,1.1mmol,1.1equiv), and DIEA (692 μL, 4 mmol, 4 175.6 (CO H), 171.6 (CONH), 165.3, 161.4 (CO-Coum), 159.1 2 equiv), compound 39 (0.168 g) was obtained after purification by (NHCO2), 152.7, 152.3, 147.7 (Cq-Ar), 138.3 (Cq-Ar), 129.6 (2 × column chromatography on silica gel (CH Cl /MeOH/AcOH: 85/ CH-Ar), 129.1(CH-Ar), 128.8 (3 × CH-Ar), 123.3 (CH-Ar), 118.7 2 2 (Cq-Ar), 109.2 (CH-Ar), 108.8 (Cq-Ar), 67.5 (OCH -Ph), 51.0 (C- 15/1) as a white powder in a 43% yield, according to general 2 procedure B. mp: 214−215 °C; 1H NMR (500 MHz, (CD ) SO): δ α), 50.5, 50.4 (CH2-N), 41.5 (CH2-Coum), 40.1 (C-ε), 32.0 (C-β), 3 2 30.8 (C-δ), 28.8 (CH ), 24.1 (C-γ), 22.5, 21.6, 21.4 (CH ); HPLC 11.94 (brs, 1H, CO H), 8.51 (d, J = 7.5 Hz, 1H, H-Ar), 7.80 (d, J = 2 2 2 8.5 Hz, 1H, NH), 7.50 (t, J = 7.5 Hz, 1H, H-Ar), 7.35−7.21 (m, 7H, (method A-2): tR = 17.0 min; HPLC-MS (method B): tR = 15.0 min; − − H-Ar, H-Ph), 7.17 (t, J = 7.5 Hz, 1H, H-Ar), 6.53 (s, 1H, H-Ar), 4.98 MS: (ESI ), m/z (%): [M − H] = 560.5 (100%), [M − C H O − − − 7 8 − (s, 2H, CH -Ph), 4.27 (m, 1H, H-α), 2.99 (m, 2H, H-ε), 1.85, 1.64 H] = 452.4 (30%); HRMS: (ESI ) calcd for [C H N O − H] : 2 31 35 3 7 8243 https://dx.doi.org/10.1021/acs.jmedchem.9b02119 J. Med. Chem. 2020, 63, 8231−8249