Notes
J . Org. Chem., Vol. 65, No. 15, 2000 4765
(m, 1H), 4.51 (m, 1H), 4.60-4.63 (m, 1H), 4.86-4.94 (m, 2H),
5.09-5.16 (m, 2H), 5.40-5.42 and 7.41-7.48 (m, 1H), 6.48-6.50
(m, 1H), 7.02-7.40 (m, 13H); 13C NMR δ 15.9, 21.3, 21.7, 23.1,
24.9, 28.2 (overlap), 32.0, 37.3, 43.5, 46.6, 52.7, 55.7, 56.7, 66.5,
69.2, 79.8, 116.2, 121.3, 123.3 and 123.5, 127.9, 128.4, 128.8 and
135.7, 151.1, 168.7 (overlap), 171.0, 171.5, 172.0; IR (KBr) 3298,
(0.0534 g, 99%) as a white solid: Rf 0.05 (10:90 MeOH/CH2Cl2);
1H NMR δ 0.82-0.98 (m, 6H), 1.26-1.41 (m, 1H), 1.44-1.45 (d,
J ) 6.68 Hz, 3H), 1.61-1.77 (m, 2H), 1.96-2.18 (m, 8H), 2.19-
2.27 (m, 2H), 2.31-2.45 (m, 2H), 2.94 (s, 3H), 3.550-3.71 (m,
2H), 4.21-4.27 (m, 1H), 4.37-4.43 (m, 1H), 4.86-4.88 (m, 1H),
5.07-5.10 (m, 1H), 5.14-5.22 (m, 1H), 6.68(s, 1H) and 7.29 (s,
1H), 7.53 (s, 1H), 8.02 (s, 1H); 13C NMR δ 15.7, 21.1, 23.2, 25.0,
25.1, 27.0, 28.4, 29.4, 31.0, 31.3, 37.1, 46.9, 52.1, 54.6, 56.8, 57.6,
69.3, 169.1, 171.3, 172.4, 173.0, 174.0, 176.9, 179.8; IR (KBr)
3300, 1734, 1654 cm-1; HRMS (EI) m/z calcd for C25H39N5O9Na
(M + Na+) 576.2645, found 576.2645.
1741, 1713, 1651 cm-1; HRMS (EI) m/z calcd for C45H56N4O10
-
Na (M + Na+) 835.3894, found 835.3921; [R]20 -23.3 (c 0.69,
D
CHCl3). Anal. Calcd for C45H56N4O10: C, 66.47; H, 6.95; N, 6.89.
Found: C, 66.13; H, 7.06; N, 6.44.
L-pGlu -L-Glu -O-L-lactyl-L-pr olyl-N-m eth yl-D-leu cin e Ben -
zyl Ester (13). A solution of compound 12 (0.4065 g, 0.5 mmol)
in EtOAc/MeOPh (20:1, 5 mL) was cooled to -20 °C, and gaseous
HCl was introduced at such a rate that the temperature of the
mixture was maintained between -10 and -20 °C at saturation.
After being stirred for 30 min at this temperature, the reaction
mixture was stirred at 0 °C for 1 h. The solution was then purged
with N2 for about 30 min, maintaining the temperature at 0 °C.
After the solution was concentrated, the residue was triturated
and washed by decantation with three 5.0 mL portions of tert-
butyl methyl ether/hexane (1:4). The product was collected by
filtration and dried in vacuo to provide the hydrochloride salt
(0.285 g, quantitative yield) as a white solid, which was used
directly in the next step. The crude hydrochloride salt was
dissolved in fresh CH2Cl2 (2 mL) at 0 °C, followed by the addition
of DIEA (0.35 mL, 2 mmol) and Z-pGluOPFP (0.215 g, 0.5 mmol).
The reaction was stirred at 0 °C for 1 h, then at room
temperature for another 1 h. The reaction mixture was quenched
with saturated NaCl solution (3 mL), and diluted with CH2Cl2
(5 mL). The mixture was separated and the aqueous layer was
extracted with CH2Cl2, the combined organic layers were washed
with 10% HCl, 5% NaHCO3 and brine, then dried (Na2SO4),
filtered and concentrated. The crude residue was purified by
flash column chromatography eluting with MeOH/CH2Cl2 (5:95)
to provide a white solid (13, 0.2614 g, 67% yield, two steps
overall): Rf 0.47 (10:90 MeOH/CH2Cl2); 1H NMR δ 0.71-0.96
(m, 6H), 1.17-1.23 (m, 1H), 1.46-1.48 (d, J ) 6.84 Hz, 3H),
1.68-1.75 (m, 2H), 1.92-2.25 (m, 10H), 2.35-2.41 (m, 2H),
2.60-2.70 (m, 1H) and 2.95 (s, 2H), 3.55-3.69 (m, 2H), 4.05-
4.10 (dd, J 1)14.27 Hz, J 2)7.13 Hz, 1H), 4.41-4.50 (m, 1H),
4.54-4.59 (m, 1H), 4.82-4.92 (m, 1H), 5.04-5.13 (m, 1H), 5.15-
5.24 (m, 4H), 5.54 (s, 1H) and 6.90 (s, 1H), 7.25-7.34 (m, 10H),
7.58-7.60 (d, J ) 6.4 Hz, 1H); 13C NMR δ 15.8, 21.1, 22.4, 23.1,
25.1, 27.2, 28.2, 31.1, 31.7, 37.4, 46.7, 52.0, 55.1, 57.0, 59.5, 66.7,
68.2, 69.2, 127.9, 128.0, 128.2, 128.4, 128.5, 128.6 and 135.1,
135.5, 151.3, 168.6, 170.7, 171.2, 171.5, 172.0, 173.5, 175.7; IR
(KBr) 3333, 3211, 1791, 1741, 1661 cm-1; HRMS (EI) m/z calcd
for C40H51N5O11Na (M + Na+) 800.3483, found 800.3513.
L-p Glu -L-Glu -O-L-la ctyl-L-p r olyl-N-m eth yl-D-leu cin e (5).
To the suspension of 10% Pd/C (0.023 g) in EtOAc/MeOH (1:1,
2 mL) was added a solution of compound 13 (0.0755 g, 0.097
mmol) in EtOAc/MeOH (1:1, 2 mL). The reaction mixture was
shaken in a Parr hydrogenator under an atmosphere of H2 for
4 h and then was filtered through a fine funnel to collect Pd/C.
The residue was washed with MeOH, and the filtrate was
concentrated in vacuo. The resulting product (5) was obtained
[(2S)-Hiv2]Did em n in M (1). To a mixture of the macrocycle
amine salt (6, 84 mg, 0.11 mmol) and side chain (5) (53.4 mg,
0.096 mmol) in CH2Cl2 (0.30 mL) at 0 °C was added BOP (51
mg, 0.12 mmol) and NMM (0.042 mL, 0.39 mmol). After 30 min
at 0 °C, the reaction mixture was allowed to warm to room
temperature and stir overnight. The solution was then treated
with saturated aqueous NaCl (2 mL) and extracted with EtOAc.
The organic layer was washed with 10% aqueous HCl, 5%
aqueous NaHCO3, and saturated aqueous NaCl, dried (Na2SO4),
filtered, and concentrated. The crude residue was purified by
flash column chromatography (the silica gel was pretreated with
ammonia washed hexane to neutral) eluting with MeOH/CHCl3
(2:98 to 10:90) to obtain 1 (0.0331 g, 32% yield) as a pale yellow
solid: Rf 0.27 (10:90 MeOH/CHCl3); 1H NMR δ 0.78-1.04 (m,
24H), 1.11-1.74 (m, 15H), 1.91-2.30 (m, 17H), 2.34-2.45 (m,
2H), 2.54 (s, 3H), 2.99 (s, 3H), 3.06-3.15 (m, 2H), 3.54-3.61 (m,
4H), 3.64-3.67 (m, 1H), 3.76 (s, 3H), 3.82-3.84 (m, 1H), 3.85-
3.91 (m, 1H), 4.13-4.16 (m, 1H), 4.22-4.24 (m, 1H), 4.46-4.50
(m, 1H), 4.56-4.60 (m, 1H), 4.70-4.74 (m, 1H), 4.84-4.88 (m,
1H), 4.93-4.94 (d, J ) 5.29 Hz, 1H), 4.98-5.03 (m, 1H), 5.11-
5.17 (m, 1H), 5.24-5.27 (m, 1H), 6.81-6.82 (d, J ) 8.57 Hz, 2H),
7.05-7.06 (d, J ) 8.51 Hz, 2H), 7.29-7.31 (m, 1H), 7.42-7.45
(m, 1H), 7.50-7.55 (m, 1H), 7.62-7.66 (m, 1H), 7.79-7.81 (d, J
) 9.83 Hz, 1H), 8.33-8.34 (d, J ) 6.66 Hz, 1H); 13C NMR δ 11.8,
14.1, 16.0, 17.9, 18.8, 20.8, 21.3, 23.5, 23.7, 24.6, 24.8 and 24.9
(overlap), 25.5, 25.9, 27.2 (overlap), 27.9 (overlap), 28.8, 29.3,
30.1 (overlap), 31.1, 33.6, 34.1, 36.0, 37.3, 38.6, 39.4, 46.8, 47.1,
48.3, 51.8, 54.5, 55.3, 56.5, 56.6, 56.7, 57.0, 66.0, 67.8, 68.7, 69.4,
71.2, 79.4, 114.1, 130.3, 132.1, 158.6, 168.5, 169.5, 170.4, 170.6,
170.7, 171.1, 171.3, 172.5 (overlap), 173.1, 174.0, 176.1, 178.6;
IR (KBr, CHCl3) 3335, 1742, 1641 cm-1; HRMS (EI) m/z calcd
for C64H98N10O18Na (M + Na+) 1317.6958, found 1317.6896;
[R]20 -47.4 (c 3.96, CHCl3).
D
Ack n ow led gm en t. We gratefully acknowledge the
financial support of the National Institutes of Health
(CA-40081), the National Science Foundation (CHE 99-
01449), and the University of Pennsylvania.
Su p p or tin g In for m a tion Ava ila ble: Copies of 1H and 13C
NMR and IR spectra for all new compounds. This material is
J O000253A