458 Journal of Medicinal Chemistry, 2010, Vol. 53, No. 1
Ayral-Kaloustian et al.
(30 mL), then dried in vacuo at 50 ꢀC for 24 h. 8-((1S,9S,
9aS,11R,11bR)-11-Acetoxy-1-(methoxymethyl)-9a,11b-dime-
thyl-3,6-dioxo-3,6,6b,7,8,9,9a,10,11,11b-decahydro-1H-furo-
[4,3,2-de]indeno[4,5-h]isochromen-9-yloxy)-8-oxooctanoic
acid, 2d, was obtained as a white powder (5.0 g, 97%). MS (ESI)
11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetra-
cosahydro-3H-23,27-epoxypyrido[2,1-c][1,4]oxazacyclohentri-
acontin-3-yl]propyl}cyclohexyl Octanedioate (8a). Following
the procedure above for the synthesis of 5a, reaction of 2d
(0.92 g, 1.57 mmol) with 31-trimethylsilyl 41-desmethoxyra-
pamycin (4b, 1.20 g, 1.26 mmol, prepared according to the
procedure for 3b)23 provided the 31-trimethylsilyl intermedi-
ate. Hydrolysis of the 31-trimethylsilyl ether, followed by
workup and silica gel purification as before, provided the
desired intermediate 6 as a white foam (1.54 g, 84%). HRMS
(ESI) m/z calcd for C81H113NO22 ([M þ Na]þ) 1474.7646,
found 1474.7646. HPLC analysis showed 98% purity (B þ C
isomers).
Diallylamine (64 mg, 0.66 mmol) was added to an ice-cold
solution of 6 (552 mg, 0.38 mmol) in TBME (15 mL). The
mixture was then stirred at 0 ꢀC for 40 h. The resulting mixture
was concentrated to about 5 mL in vacuo and then triturated
with hexane (30 mL). The product 8a was collected on a Buchner
funnel as a yellow powder (540 mg, 90%). HRMS (ESI) m/z
calcd for C87H124N2O22 ([M þ Na]þ) 1571.8538, found
1571.8532; 1H NMR (CDCl3, 300 MHz, representative data) δ
8.15 (s, 1H), 6.82 (br, 1H), 6.44-5.83 (m, 6H), 5.56-5.10 (m,
9H), 4.86-4.16 (m, 5H), 3.97-3.11 (m, 23H), 2.80-2.24 (m,
13H); 13C NMR (CDCl3, 100 MHz, representative data) δ
216.0, 207.6, 192.5, 179.0, 173.5, 173.2, 169.8, 169.3, 165.8,
150.6, 140.7, 139.3, 137.6, 136.1, 135.8, 133.4, 130.0, 129.3,
127.1, 126.6, 119.4, 98.5, 89.2, 84.9, 84.4, 82.5, 80.2, 77.2, 75.8,
73.1, 73.0, 69.3, 67.2, 59.4, 59.3, 55.9, 51.3, 49.4, 46.6, 44.4, 44.2,
43.0, 42.8, 41.5, 41.4, 40.8, 40.2, 38.7, 38.5, 35.1, 34.6, 34.3, 33.8,
33.7, 33.2, 32.0, 31.6, 31.4, 31.3, 30.3, 28.8, 28.7, 27.8, 27.3, 27.0,
27.0, 25.3, 24.8, 24.3, 21.5, 21.2, 20.6, 16.3, 16.0, 15.8, 13.8, 13.4,
13.1, 12.8, 10.2.
1E,4S,4aR,5R,6aS,7S)-5-Acetyloxy-1-({[3-(dimethylamino)-
propyl](methyl)amino}methylene)-11-hydroxy-4-(methoxymethyl)-
4a,6a-dimethyl-2,10-dioxo-1,2,4,4a,5,6,6a,7,8,9,9a,10-dodecahydro-
indeno[4,5-h]isochromen-7-yl-trans-4-{(2R)-2-[(3S,6R,7E,9R,10R,
12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,27-dihydroxy-
10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-1,5,11,28,29-penta-
oxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,
34,34a-tetracosahydro-3H-23,27-epoxypyrido[2,1-c][1,4]oxaza-
cyclohentriacontin-3-yl]propyl}cyclohexyl Octanedioate (8b). A
solution of 41-desmethoxyrapamycin-suberate-wortmannin
conjugate 6 (552 mg, 0.38 mmol) in TBME (15 mL) was cooled
to -30 ꢀC. N,N,N0-Trimethyl-1,3-propanediamine (51 mg, 0.44
mmol) in TBME (3 mL) was added dropwise over 10 min. After
addition, the mixture was stirred at -30 ꢀC for 1 h, then warmed
to -20 ꢀC and stirred for another 1 h. Hexane (20 mL) was
introduced while maintaining the temperature at -15 to -20
ꢀC. The product 8b was collected on a Buchner funnel as a
yellow powder (550 mg, 92%). HRMS (ESI) m/z calcd for
C87H130N3O22 ([M þ H]þ) 1568.9140, found 1568.9128; 1H
NMR (CDCl3, 300 MHz, representative data) δ 8.15 (s, 1H),
6.44-5.85 (m, 5H), 5.56-5.11 (m, 4H), 4.86-4.16 (m, 5H),
3.88-3.10 (m, 23H), 2.80-2.22 (m, 28H); 13C NMR (CDCl3,
100 MHz, representative data) δ 215.3, 208.3, 192.6, 179.1,
173.6, 173.2, 169.9, 169.2, 166.8, 149.9, 140.1, 138.6, 137.7,
136.1, 135.6, 133.6, 130.2, 129.6, 126.6, 126.4, 98.5, 88.3, 84.9,
84.3, 82.6, 80.3, 77.2, 75.7, 73.2, 73.0, 69.2, 67.1, 59.4, 59.4, 59.2,
58.5, 55.9, 51.3, 46.6, 45.3, 44.4, 44.2, 43.0, 42.7, 41.6, 41.5, 40.8,
40.2, 38.9, 38.5, 35.1, 34.6, 34.3, 33.8, 33.7, 33.2, 32.0, 31.6, 31.4,
31.2, 30.3, 28.8, 28.7, 27.8, 27.3, 27.0, 25.7, 25.3, 24.8, 24.3, 21.5,
21.2, 20.6, 16.3, 16.0, 15.9, 15.8, 13.8, 13.4, 13.1, 10.2.
1
m/z 609 (M þ Na); H NMR (CDCl3, 300 MHz) δ 10.35 (br,
1H), 8.19 (s, 1H), 6.08-6.03 (m,1H), 4.82-4.74 (m, 2H),
3.45-3.41 (m, 1H), 3.15 (s, 3H), 2.98-2.94 (m, 1H), 2.8-2.77
(m, 1H), 2.61-2.56 (m, 1H), 2.50-2.43 (m, 1H), 2.34-2.26 (m,
4H), 2.10 (s, 3H), 2.07-2.05 (m, 2H), 1.74-1.33 (m, 14H), 0.85
(s, 3H); 13C NMR (CDCl3, 75 MHz) δ 179.72, 174.07, 173.19,
170.12, 158.10, 150.32, 148.96, 145.28, 143.03, 141.67, 114.51,
89.25, 79.99, 73.21, 70.80, 59.75, 45.04, 44.40, 41.02, 40.52,
34.61, 34.24, 29.06, 29.00, 27.85, 26.89, 25.10, 25.05, 24.82,
21.49, 13.22.
Reaction of 2d (5.86 g, 10 mmol), with 3b (7.89 g, 8 mmol),
followed by hydrolysis of the intermediate trimethylsilyl
ether, according to the procedure above for 5a, gave the desired
product 5c as a white foam (8.5 g, 72%). HRMS (ESI) m/z calcd
for C82H115NO23 ([M þ Na]þ) 1504.7752, found 1504.7755.
HPLC analysis showed >99% purity (B þ C isomers).
A solution of 5c (7.80 g, 5.26 mmol) in TBME (250 mL) was
treated with N,N,N0-trimethyl-1,3-propanediamine (700 mg,
6 mmol) as before for 7a, to yield 7c as a yellow powder (7.95
g, 94%). HRMS (ESI) m/z calcd for C88H132N3O23 ([M þ H]þ)
1598.9246, found 1598.9234; 1H NMR (CDCl3, 300 MHz,
representative data) δ 8.14 (s, 1H), 6.43-5.87 (m, 5H), 5.59-
5.11 (m, 4H), 4.86-4.18 (m, 5H), 3.90-3.11 (m, 27H); 2.75-
2.23 (m, 24H); 13C NMR (CDCl3, 100 MHz, representative
data) δ 215.0, 208.2, 192.8, 179.1, 173.6, 173.2, 169.9, 169.3,
166.7, 150.0, 140.0, 137.7, 136.1, 135.7, 133.5, 130.2, 129.4,
126.5, 98.5, 88.3, 84.8, 84.3, 82.6, 80.9, 80.3, 77.3, 76.0, 75.5,
73.2, 69.2, 67.2, 59.4, 59.2, 58.5, 57.5, 56.0, 51.3, 49.4, 46.6, 45.3,
44.4, 44.2, 43.0, 42.0, 41.6, 41.5, 40.6, 40.2, 39.0, 38.4, 36.0, 35.1,
35.0, 34.3, 33.8, 33.3, 32.8, 31.3, 31.2, 29.8, 28.8, 28.7, 27.8, 27.2,
27.0, 25.7, 25.3, 24.8, 24.8, 24.3, 21.5, 21.2, 20.7, 16.2, 16.0, 15.9,
13.7, 13.4, 13.2, 10.2.
(1E,4S,4aR,5R,6aS,7S)-5-Acetyloxy-1-[(diallylamino)methyl-
ene]-11-hydroxy-4-(methoxymethyl)-4a,6a-dimethyl-2,10-dioxo-
1,2,4,4a,5,6,6a,7,8,9,9a,10-dodecahydroindeno[4,5-h]isochromen-
7-yl-(1R,2R,4S)-4-{(2R)-2-[(3S,6R,7E,9R,10R,12R,14S,15E,17E,
19E,21S,23S,26R,27R,34aS)-9,27-dihydroxy-10,21-dimethoxy-6,8,
12,14,20,26-hexamethyl-1,5,11,28,29-pentaoxo-1,4,5,6,9,10,11,12,13,
14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetracosahydro-3H-
23,27-epoxypyrido[2,1-c][1,4]oxazacyclohentriacontin-3-yl]pro-
pyl}-2-methoxycyclohexyl Octanedioate (7d). A solution of 5c
(1.0 g, 0.67 mmol) in TBME (30 mL) was cooled in an ice bath
and treated with diallylamine (0.15 mL) for 48 h and then
concentrated to a volume of about 10 mL and triturated with
hexane (50 mL). The product (7d) was collected on a Buchner
funnel as a yellow powder (985 mg, 92%). HRMS (ESI) m/z
calcd for C88H126N2O23 ([M þ Na]þ) 1601.8644, found
1
1601.8637; H NMR (CDCl3, 300 MHz, representative data)
δ 8.14 (s, 1H), 6.82 (br, 1H), 6.43-5.83 (m, 6H), 5.58-5.10 (m,
9H), 4.86-4.18 (m, 5H), 3.96-3.11 (m, 26H), 2.75-2.28 (m,
13H), 2.10-0.81 (m, 65H); 13C NMR (CDCl3, 100 MHz,
representative data) δ 215.0, 208.2, 192.8, 179.0, 173.6, 173.2,
169.8, 169.3, 166.8, 150.6, 139.9, 139.4, 137.5, 136.1, 135.8,
133.5, 130.2, 129.4, 126.4, 119.4, 98.5, 89.1, 84.8, 84.2, 82.5,
80.9, 80.2, 77.1, 76.0, 75.4, 73.1, 69.3, 67.2, 59.4, 59.1, 57.5,
55.9, 55.9, 51.3, 49.4, 46.6, 44.4, 44.2, 43.0, 42.7, 41.5, 40.6,
40.2, 39.0, 38.4, 36.0, 35.1, 34.6, 34.3, 33.8, 33.3, 32.9, 31.3,
31.2, 29.8, 28.8, 28.7, 27.8, 27.2, 27.0, 25.3, 24.8, 24.3, 21.4,
21.2, 20.7, 16.2, 16.0, 15.9, 13.6, 13.4, 13.3, 10.2.
(1E,4S,4aR,5R,6aS,7S)-5-Acetyloxy-1-[(diallylamino)methyl-
ene]-11-hydroxy-4-(methoxymethyl)-4a,6a-dimethyl-2,10-dioxo-
1,2,4,4a,5,6,6a,7,8,9,9a,10-dodecahydroindeno[4,5-h]isochromen-
7-yl-trans-4-{(2R)-2-[(3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,
21S,23S,26R,27R,34aS)-9,27-dihydroxy-10,21-dimethoxy-6,8,
12,14,20,26-hexamethyl-1,5,11,28,29-pentaoxo-1,4,5,6,9,10,
Acknowledgment. We thank George Morton for NMR
data, Dr. Xidong Feng for HRMS data, Dr. Mark Tischler
for solubility analyses, and Jessica Lucas for biomarker studies.
Supporting Information Available:
Additional details of
experimental procedures, representative 1H, 13C NMR, MS,
and HPLC traces, and additional methods for stability studies.