1262
H. Singh, R. Warmuth / Tetrahedron 58 -2002) 1257±1264
yield). Mp 130.58C; dH 3CDCl3) 7.62 31H, d, J9.6 Hz),
7.37 31H, d, J8.4 Hz), 6.84 31H, dd, J8.4, 2.4 Hz),
6.77 31H, d, J2.4 Hz), 6.24 31H, d, J9.6 Hz), 4.33 32H,
t, J6.0 Hz), 4.66 32H, t, J6.0 Hz); dC 3CDCl3) 161.39,
161.17, 143.50, 129.19, 113.73, 113.27, 113.106, 113.00,
101.91, 68.43, 28.72.
162.08, 161.94, 156.05, 148.84, 148.18, 147.60, 147.29,
147.07, 146.89, 145.22, 144.89, 144.07, 143.62, 140.02,
139.67, 139.50, 139.39, 139.21, 139.37, 139.27, 139.17,
139.01, 129.43, 115.20, 114.84, 114.74, 114.66, 113.43,
113.31, 113.24, 101.75, 99.88, 99.44, 99.12, 74.67, 73.41,
72.53, 72.26, 72.13, 72.02, 67.70, 37.32, 37.21, 32.35,
32.29, 30.54, 30.05, 29.59, 27.90, 27.83, 27.79, 22.97,
22.95, 22.91, 14.39. MALDI HRMS m/z 2418.045
3M1Na1) 3calcd for C138H162NaO36, 2418.074).
4.1.3. Compound 5. Host 3 3100 mg, 0.047 mmol)
was dissolved under argon in degassed DMF 315 mL).
Anhydrous K2CO3 32 g) was added and the suspension
was stirred for 1 h. Linker 2 338 mg, 0.14 mmol) was
added to the suspension and stirring was continued at
room temperature for 3 days. The suspension was ®ltered
and the ®ltrate poured into phosphate buffer 30.2 M, pH 6,
40 mL). The precipitated crude product was ®ltered off and
washed with water 32£10 mL) and CH3OH 32£10 mL). It
was redissolved in the minimum amount of CHCl3 and
precipitated again by adding 10 volume equivalents of
CH3OH. After ®ltering off the crude product it was puri®ed
by column chromatography on silica gel using CH2Cl2 as
the mobile phase. Concentration of the product fraction
gave 5 as a white solid 357 mg, 50% yield). Mp 180±
1818C; dH 3CDCl3) 7.63 32H, d, J9.6 Hz), 7.43 32H, d,
J8.8 Hz), 7.00 32H, dd, J8.8, 1.6 Hz), 6.88 32H, d, J
1.6 Hz), 6.80 32H, s), 6.79 34H, s), 6.77 32H, s), 6.27 32H, d,
J7.6 Hz), 6.24 32H, d, J9.6 Hz), 5.82 32H, d, J8.0 Hz),
5.80 32H, d, J8.0 Hz), 5.55 32H, d, J7.2 Hz), 4.82±4.72
32H, m), 4.72±4.54 312H, m), 4.52±4.46 32H, m), 4.40±
4.31 32H, m), 4.30±4.07 312H, m), 4.05±3.96 36H, m),
3.52±3.40 36H, m), 2.22±2.08 316H, m), 1.48±1.26 366H,
m), 0.95±0.88 324H, m). dC 3CDCl3) 162.20, 161.45,
156.23, 149.13, 149.05, 148.79, 148.68, 148.61, 148.42,
148.27, 148.14, 144.40, 144.35, 144.25, 143.67, 143.54,
139.42, 139.39, 139.25, 139.21, 139.13, 139.05, 138.99,
129.33, 115.19, 115.03, 114.94, 114.85, 113.48, 113.12,
113.01, 110.45, 110.35, 101.83, 100.69, 100.02, 99.77,
99.58, 78.81, 78.55, 78.37, 76.39, 75.97, 75.41, 72.16,
67.38, 37.34, 37.28, 32.48, 32.42, 30.61, 30.25, 29.88,
28.20, 28.12, 27.98, 22.99, 22.94, 14.43. MALDI HRMS
m/z 2458.113 320%) 3M22£H2CCCH21Na1) 3calcd for
C141H166NaO36, 2458.106); 2418.116 3100%) 3M23£
H2CCCH21Na1) 3calcd for C138H162NaO36, 2418.074).
Anal. calcd for C147H174O36: C, 70.15; H, 6.96. Found: C,
70.09; H, 6.67.
4.1.5. Compound 7. Host 6 350 mg, 0.021 mol) was
dissolved in dry THF 315 mL). Subsequently, N-BOC-
glycine 363 mg, 0.624 mmol), p-dimethylaminopyridine
33 mg, 0.024 mmol) and 1-[3-3dimethylamino)propyl]-3-
ethylcarbodiimide methiodide 3191 mg, 0.643 mmol) were
added. The reaction mixture was stirred under argon at room
temperature for 5 days. The formed precipitate was ®ltered
off. The ®ltrate was concentrated under reduced pressure.
The remaining crude product was washed with water
32£20 mL) and was puri®ed via semi-preparative HPLC
3Luna-SiO2 column; 10m, 10£250 mm2 3Phenomenex);
CH2Cl2/THF 395:5); ¯ow rate 10 mL/min; detection at
254 nm; retention time of 735 min). The product fractions
were concentrated down to 1 mL. The product was precipi-
tated by the addition of hexanes 310 mL) and was ®ltered
and dried at high vacuum to yield 7 as a white solid 345 mg,
65% yield). Mp 183±1848C 3decomp.); dH 3CDCl3; 333 K)
7.65 32H, d, J9.6 Hz, coumarin H3), 7.42 32H, d, J
8.4 Hz, coumarin H5), 6.88 32H, dd, J8.4, 2.0 Hz,
coumarin H6), 6.80 34H, s, ArH), 6.79 32H, s, ArH), 6.78
32H, s, ArH), 6.76 32H, d, J2.0 Hz, coumarin H8), 6.22
32H, d, J9.6 Hz, coumarin H4), 6.24 32H, d, J9.6 Hz),
6.13±5.97 34H, m, OCHHouterO), 5.94 32H, d, J7.2 Hz,
OCHHouterO), 5.75 32H, d, J6.8 Hz, OCHHouterO), 5.66±
5.42 36H, m, NH), 4.76±4.67 36H, m, CH,CH2,CH2), 4.56
32H, t, J7.8 Hz, CH,CH2,CH2), 4.46±4.16 324H, m,
OCH2CH2O, OCHHinnerO, OCH2CH), 4.14 32H, d, J
7.2 Hz, OCHHinnerO), 4.13±4.02 32H, m, OCH2CH),
4.0±3.6 312H, m, NHCH2), 3.82, 3.73 36H, AB-
system, J18.2 Hz, OCH2CH), 2.25±2.08 316H, m,
CH2CH2CH2CH2CH3), 1.48 318H, s, C3CH3)3) 1.46 336H,
s C3CH3)3), 1.48±1.26 348H, m, CH2CH2CH2CH2CH3),
0.95±0.88 324H, m, CH2CH2CH2CH2CH3). dC 3CDCl3,
335 K) 169.97, 169.83, 169.75, 162.29, 161.17, 156.36,
156.16, 156.11, 155.97, 148.80, 148.60, 148.31, 148.20,
148.13, 148.04, 144.08, 143.55, 139.65, 139.39, 139.35,
139.23, 139.21, 139.15, 129.33, 115.14, 115.06, 114.95,
114.87, 113.63, 113.29, 113.15, 102.14, 99.68, 99.64,
99.54 3sb), 99.38, 80.48, 80.34, 80.24, 72.72, 71.74, 70.19,
70.11 3sb), 67.92, 43.11, 43.03, 37.37, 32.41, 32.37, 32.34,
30.49, 30.39, 30.32, 29.97, 28.73, 28.02, 27.98, 27.96,
27.93, 22.94, 22.95, 22.92, 14.27. MALDI HRMS m/z
3360.486 3M1Na1) 3calcd for C180H228N6NaO54,
3360.517). Anal. calcd for C180H228N6O54: C, 64.73; H,
6.88; N, 2.52. Found: C, 65.05; H, 6.97; N, 2.86.
4.1.4. Compound 6. Host 5 330 mg, 0.0123 mmol) was
dissolved in
a solvent mixture containing CD3OD
3300 mL), CDCl3 3100 mL), D2O 350 mL), and CF3COOH
320 mL) in a NMR tube. The tube was sealed with a Te¯on
screw-cap and the solution was heated to 608C in an oil bath.
Periodically, the progress of the reaction was checked by
recording a 1H NMR spectrum. The reaction was complete
after 10 days. The solvent was removed by evaporation to
give 6 329.5 mg, quantitative) as a colorless solid. Mp 2688C
3decomp.); dH 3CDCl3) 7.64 32H, d, J9.6 Hz), 7.43 32H, d,
J8.8 Hz), 7.00 32H, dd, J8.4, 2.0 Hz), 6.89 32H, d,
J2.0 Hz), 6.80 32H, s), 6.79 34H, s), 6.77 32H, s), 6.27
32H, d, J7.6 Hz), 6.24 32H, d, J9.6 Hz), 5.82 32H, d,
J8.0 Hz), 5.80 32H, d, J8.0 Hz), 5.55 32H, d, J
7.2 Hz), 4.82±4.72 32H, m), 4.72±4.54 312H, m), 4.52±
4.46 32H, m), 4.40±4.31 32H, m), 4.30±4.07 312H, m),
4.05±3.96 36H, m), 3.52±3.40 36H, m), 2.22±2.08 316H,
m), 1.48±1.26 366H, m), 0.95±0.88 324H, m). dC 3CDCl3)
4.1.6. Compound 1. Host 7 35 mg) was placed into a 5 mL
round bottom ¯ask. Ice-cold CF3COOH 30.2 mL) was added
at 08C. The suspension was stirred until all of 7 had
dissolved and was left at 08C for 30 min. The solvent was
removed under vacuum. CHCl3 31 mL) was added and was
evaporated to leave 1 as a white powder, which was dried at
high vacuum. Mp 3208C 3decomp.); dH 3D2O/CD3OD/