Synthesis and biological activity of novel epothilone aziridines.

Article abstract of DOI:10.1021/ol016273w

[reaction: see text]. A series of 12alpha,13alpha-aziridinyl epothilone derivatives were synthesized in an efficient manner from epothilone A. The final semisynthetic route involves a formal double-inversion of stereochemistry at both the C12 and C13 posi

Full text of DOI:10.1021/ol016273w

Supporting Information
Synthesis and Biological Activity of Novel Epothilone Aziridines
Alicia Regueiro-Ren,* Robert M. Borzilleri, Xiaoping Zheng, Soong-Hoon Kim,
James A. Johnson, Craig R. Fairchild, Francis Y. F. Lee, Byron H. Long, Gregory D. Vite
Divisions of Discovery Chemistry and Oncology Drug Discovery, The Bristol-Myers Squibb
Pharmaceutical Research Institute, P.O. Box 4000, Princeton, NJ 08543-4000 (USA).
[4S-[4R*,7S*,8R*,9R*,13S*,14S*,16R*(E)]]-14-Bromo-4,8-bistriethylsilyloxy-13-hydroxy-5,5,7,9-
tetramethyl-16-[1-methyl-2-(2-methyl- 4-thiazolyl)ethenyl]-1-oxacyclohexadecane-2,6-dione (8).
OH
Br
S
N
OTES
O
O
O
OTES
Et3SiCl (25 mL, 149 mmol) was added to epothilone A (10.39 g, 21 mmol), N,N-diisopropylethylamine (55 mL,
315 mmol), and imidazole (7.15 g, 105 mmol) in DMF (75 mL) at 25 oC. The reaction mixture was heated at 55 °C for 6.5 h
and concentrated in vacuo. The residue was then diluted with CH2Cl2 (100 mL) and the organic extracts were washed with
aqueous NaHCO3 (30 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by flash chromatography
(SiO2, 5.0 x 30 cm, hexanes to 15% EtOAc/hexanes gradient elution) to afford bis-TES-epothilone A as a white solid (15.1
g, >95%). MS (ESI+): (M+H)+ 722. To a solution of bis-TES-epothilone A (2.0 g, 2.8 mmol) in CH2Cl2 (30 mL) at –20 oC
under Ar was added MgBr2•OEt2 (3 x 1.1 g, 12 mmol total) in three portions every two hours while maintaining an internal
temperature between –15 and –5 oC. After 7 h, the reaction mixture was quenched with pH 7 aqueous phosphate buffer (40
mL) and brine (40 mL), carefully extracted with EtOAc (3 x 100 mL), dried (Na2SO4), and concentrated in vacuo. The
residue was purified by flash chromatography (SiO2, 4.5 x 25 cm, 10-20 % EtOAc/hexanes gradient elution) to afford 8 as a
white solid [1.0 g, 45 % (67 % based on 0.6 g of recovered starting material; <2 % of the other C13-Br/C14-OH regioisomer
1
was detected]. H NMR (400 MHz, CDCl3) δ 6.97 (s, 1 H), 6.55 (s, 1 H), 5.48-5.46 (m, 1 H), 4.39-4.38 (m, 1 H), 3.69-3.66
(m, 1 H), 3.01-2.99 (m, 1 H), 2.76-2.74 (m, 1 H), 2.73 (s, 3 H), 2.71-2.56 (m, 3 H), 2.49-2.47 (m, 1 H), 2.13 (s, 3 H), 1.63-
1.61 (m, 4 H), 1.30-1.28 (m, 2 H), 1.23 (s, 3 H), 1.14 (s, 3 H), 0.99-0.93 (m, 26 H), 0.72-0.63 (m, 12 H). MS (ESI+): (M+H)+
802.
[4S-[4R*,7S*,8R*,9R*,13S*,14R*,16R*(E)]]-14-Azido-4,8-bistriethylsilyloxy-13-hydroxy-5,5,7,9-tetramethyl-
16-[1-methyl-2-(2-methyl- 4-thiazolyl)ethenyl]-1-oxacyclohexadecane-2,6-dione (9).
OH
N3
S
N
OTES
O
O
O
OTES
To a solution of 8 (0.17 g, 0.21 mmol) in DMF (2 mL) under Ar was added sodium azide (0.14 g, 2.1 mmol) and the
o
resulting suspension was warmed to 43 C. After 36 h, the solvent was removed in vacuo and the residue was directly
purified by flash chromatography (SiO2, 2.5 x 15 cm, 10-20 % EtOAc/hexanes gradient elution) to give 9 (0.14 g, 88 %) as a
1
white foam. H NMR (400 MHz, CDCl3) δ 6.91 (s, 1 H), 6.51 (s, 1 H), 5.26-5.23 (m, 1 H), 3.99-3.92 (m, 1 H), 3.90-3.86 (m,
1 H), 3.81-3.72 (m, 1 H), 3.41-3.37 (m, 1 H), 2.92-2.89 (m, 1 H), 2.82-2.74 (m, 1 H), 2.64 (s, 3 H), 2.57-2.52 (m, 1 H), 2.21-
2.18 (m, 1 H), 2.03 (s, 3 H), 1.81-1.42 (m, 5 H), 1.37-0.96 (m, 10 H), 0.92-0.75 (m, 24 H), 0.67-0.51 (m, 12 H). MS (ESI+):
(M+H)+ 765.
[4S-[4R*,7S*,8R*,9R*,13R*,14R*,16R*(E)]]-14-Azido-4,8-bistriethylsilyloxy-13-hydroxy-5,5,7,9-tetramethyl-16-[1-
methyl-2-(2-methyl- 4-thiazolyl)ethenyl]-1-oxacyclohexadecane-2,6-dione (10).

OH
N3
S
N
OTES
O
O
O
OTES
To a solution of 9 (0.10 g, 0.13 mmol) in THF under Ar was sequentially added 4-nitrobenzoic acid (55 mg, 0.33
mmol), triphenylphosphine (86 mg, 0.33 mmol), and diethyl azodicarboxylate (52 µL, 0.33 mmol). The reaction mixture
o
was stirred at 25 C for 1.5 h, concentrated in vacuo and the residue was purified by flash chromatography (SiO2, 2.5 x 10
cm, 10-20 % EtOAc/hexanes gradient elution) to afford the corresponding p-nitrobenzoate (0.10 g, 86 %) as a white foam.
MS (ESI+): 914.6 (M+H)+. This intermediate (0.10 g, 0.11 mmol) was then treated with 2.0 M ammonia in methanol (1 mL)
at 25 oC under Ar for 4 h. The solvent was removed in vacuo and the residue was directly purified by flash chromatography
(SiO2, 1.5 x 10 cm, 10-30% EtOAc/hexanes gradient elution) to afford 10 (71 mg, 85%) as a white foam. 1H NMR (400
MHz, CDCl3) δ 6.97 (s, 1 H), 6.59 (s, 1 H), 5.55-5.52 (m, 1 H), 4.42-4.39 (m, 1 H), 3.96-3.94 (m, 1 H), 3.61-3.56 (m, 1 H),
3.05- 3.01 (m, 1 H), 2.75-2.74 (m, 1 H), 2.70 (s, 3 H), 2.61-2.56 (m, 1 H), 2.13 (s, 3 H), 2.04-2.02 (m, 1 H), 1.84-1.50 (m, 3
H), 1.18 (s, 3 H), 1.14-1.10 (m, 6 H), 1.07-0.93 (m, 24 H), 0.68-0.57 (m, 12 H). MS (ESI+): 765.5 (M+H)+; MS (ESI-): 763.3
(M-H)-.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Bistriethylsilyloxy-8,8,10,12-tetramethyl-3-[1-methyl-2-(2-
methyl- 4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (11).
HN
S
N
OTES
O
O
O
OTES
To a solution of 10 (56.5 mg, 67 µmol) in THF- H2O (12:1, 1.2 mL) under Ar was added trimethylphosphine (134
µL, 1.0 M in THF). The reaction mixture was warmed to 45 oC for 10 h, and the solvent was removed under a constant flow
of nitrogen. The residue was purified by flash chromatography (SiO2, 1 x 5 cm, 0-5 % MeOH/CHCl3 gradient elution) to
1
afford 11 (41 mg, 85 %) as a film. H NMR (400 MHz, CDCl3) δ 6.94 (s, 1 H), 6.52 (s, 1 H), 5.20-5.18 (m, 1 H), 4.02-4.00
(m, 1 H), 3.91-3.89 (m, 1 H), 2.99-2.96 (m, 1 H), 2.72-2.70 (m, 2 H), 2.66 (s, 3 H), 2.17-2.14 (m, 1 H), 2.10-2.09 (m, 1 H),
2.06 (s, 3 H), 1.95-1.91 (m, 1 H), 1.69-1.27 (m, 5 H), 1.14 (s, 3 H), 1.11 (s, 3 H), 0.98-0.87 (m, 24 H), 0.66-0.54 (m, 12 H).
MS (ESI+): 537.3 (M+H)+.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-7,11-Dihydroxy-8,8,10,12-tetramethyl-3-[1-methyl-2-(2-methyl-
4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (7).
HN
S
N
OH
O
O
O
OH
Compound 11 (17.2 mg, 24 µmol) was treated with 20 % trifluoroacetic acid in methylene chloride (0.5 mL) at 0 oC
under Ar for 10 min. The reaction mixture was concentrated under a constant stream of nitrogen at 0 oC and the residue was
purified by radial chromatography (1 mm SiO2 GF rotor, 2-10 % MeOH-CHCl3 gradient elution) to afford 7 (10.6 mg, 86 %)
1
as a white solid; H NMR (400 MHz, CDCl3) δ 6.84 (s, 1 H), 6.52 (s, 1 H), 5.46-5.44 (m, 1 H), 4.08- 4.04 (m, 1 H), 3.69-
3.68 (m, 1 H), 3.20-3.17 (m, 1 H), 2.57 (s, 3 H), 2.44-2.38 (m, 1 H), 2.34-2.30 (m, 1 H), 1.95-1.93 (br s, 4 H), 1.82-1.78 (m,
2 H), 1.71-1.53 (m, 2 H), 1.46-1.31 (m, 4 H), 1.27 (s, 3 H), 1.23-1.01 (m, 2 H), 0.99 (d, 3 H, J = 6.9 Hz), 0.91 (s, 3 H), 0.81
(d, 3 H, J = 6.9 Hz); 13C NMR (300 MHz, CDCl3) δ 220.3, 171.2, 165.1, 152.4, 136.8, 119.2, 115.9, 76.5, 66.1, 75.4, 52.5,
45.0. 38.1, 34.6, 34.2, 30.1, 29.9, 26.1, 24.9, 22.9, 19.3, 17.6, 16.4, 15.4, 14.3. MS (ESI+): 493.2 (M+H)+.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-17-Methyl-7,11-dihydroxy-8,8,10,12-tetramethyl-3-[1-methyl-2-
(2-methyl- 4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (12a).

N
O
S
N
OH
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.97 (s, 1 H), 6.66 (s, 1 H), 5.72-5.61 (m, 1 H), 4.35-4.15 (m, 1 H), 3.84-3.83 (m, 1
H), 3.34-3.33 (m, 1 H), 2.72 (s, 3 H), 2.57-2.47 (m, 2 H), 2.39 (s, 3 H), 2.08 (s, 3 H), 1.96-1.85 (m, 2 H), 1.72-1.45 (m, 5 H),
1.42 (s, 3 H), 1.37-1.20 (m, 2 H), 1.18-1.16 (m, 2 H), 1.12 (d, 3 H, J = 6.9 Hz), 1.03 (s, 3 H), 0.93 (d, 3 H, J = 6.8 Hz). MS
(ESI+): 507.3 (M + H) +.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-Benzyl-7,11-dihydroxy-8,8,10,12-tetramethyl-3-[1-methyl-2-
(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (12b).
Ph
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 7.31 (s, 5 H), 6.99 (s, 1 H), 6.67 (s, 1 H), 5.50-5.47 (m, 1 H), 4.09-4.00 (m, 1 H),
3.78-3.73 (m, 2 H), 3.37-3.33 (m, 1 H), 3.20-3.17 (m, 1 H), 2.73 (s, 3 H), 2.54 (dd, 1 H, J = 13.8, 10.2 Hz), 2.46 (dd, 1 H, J
= 13.8, 3.0 Hz), 2.08 (s, 3 H), 1.98-1.84 (m, 4 H), 1.58-1.48 (m, 3 H), 1.42-1.37 (m, 4 H), 1.36 (s, 3 H), 1.28-1.22 (m, 2 H),
1.16 (d, 3 H, J = 6.8 Hz), 1.11 (s, 3 H), 0.95 (d, 3 H, J = 6.9 Hz); LRMS (ESI+): 583.4 (M + H)+.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-[2-Methylpropyl]amido-9-7,11-dihydroxy-8,8,10,12-
tetramethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (13a).
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.97 (s, 1 H), 6.57 (s, 1 H), 5.38-5.35 (m, 1 H), 4.15- 4.11 (m, 1 H), 3.84-3.80 (m, 2
H), 3.18-3.15 (m, 1 H), 2.70 (s, 3 H), 2.66-2.53 (m, 2 H), 2.52-2.49 (m, 2 H), 2.45-2.11 (m, 6 H), 2.09 (s, 3 H), 1.84-1.72 (m,
3 H), 1.55-1.40 (m, 4 H), 1.36 (s, 3 H), 1.19 (s, 3 H, J = 6.8 Hz), 1.14 (s, 3 H), 1.02 (d, 3 H, J = 6.9 Hz), 0.95-0.93 (m, 6 H).
MS (ESI+): 577.3 (M + H)+.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-[(N,N-dimethylamino)methyl]amido-7,11-dihydroxy-
8,8,10,12-tetramethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione
(13b).
N
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.97 (s, 1 H), 6.57 (s, 1 H), 5.37-5.35 (m, 1 H), 4.15-4.12 (m, 1 H), 3.83-3.81 (m, 1
H), 3.26 (br s, 2 H),3.19-3.16 (m, 1 H), 2.70 (s, 3 H), 2.58-2.41 (m, 3 H), 2.37 (s, 6 H), 2.32-2.28 (m, 1 H), 2.09 (s, 3 H),

2.07-1.80 (m, 4 H), 1.79-1.41 (m, 3 H), 1.19 (s, 3 H, J = 6.8 Hz), 1.14 (s, 3 H), 1.02 (d, 3 H, J = 7.0 Hz). MS (ESI+): 578.3
(M + H)+.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17- N-Methylamido -7,11-dihydroxy-8,8,10,12-tetramethyl-3-[1-
methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (13c).
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.99 (s, 1 H), 6.58 (s, 1 H), 5.39-5.37 (m, 1 H), 3.86-3.83 (m, 1 H), 3.75-3.73 (m, 1
H), 3.20-3.18 (m, 1 H), 2.71 (s, 3 H), 2.57-2.46 (m, 3 H), 2.42-2.38 (m, 1 H), 2.29-2.25 (m, 1 H), 2.14 (s, 3 H) 2.11 (s, 3 H),
1.87-1.74 (m, 4 H), 1.56-1.41 (m, 4 H), 1.37 (s, 3 H), 1.20 (d, 3 H, J = 6.8 Hz), 1.16 (s, 3 H), 1.03 (d, 3 H, J = 6.9 Hz). MS
(ESI+): 535.3 (M + H)+.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-Phenylamido-7,11-dihydroxy-8,8,10,12-tetramethyl-3-[1-
methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (13d).
Ph
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 7.96-7.93 (m, 2 H), 7.60-7.56 (m, 1 H), 7.49-7.45 (m, 2 H), 7.00 (s, 1 H), 6.59 (s, 1
H), 5.37 (d, 1 H, J = 8.1 Hz), 3.85-3.83 (m, 1 H), 3.72 (br s, 1 H), 3.21-3.18 (m, 1 H), 2.72 (s, 3 H), 2.69-2.43 (m, 6 H), 2.06
(s, 3 H), 2.06-1.93 (m, 4 H), 1.79-1.71 (m, 2 H), 1.64-1.45 (m, 3 H), 1.39 (s, 3 H), 1.21 (d, 3 H, J = 6.8 Hz), 1.17 (s, 3 H),
1.05 (d, 3 H, J = 6.9 Hz). MS (ESI+) 597.3 (M+ + H).
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-17- N-(2-Thiophenyl)carbamoyl -7,11-dihydroxy-8,8,10,12-
tetramethyl-3-[1-methyl-2-(2-methyl- 4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (13e).
S
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 7.76 (d, 1 H, J = 3.9 Hz), 7.58 (d, 1 H, J = 4.6 Hz), 7.14 (dd, 1 H, J = 4.3, 3.9 Hz),
7.00 (s, 1 H), 6.59 (s, 1 H), 5.40-5.37 (m, 1 H), 3.85-3.84 (m, 1 H), 3.73-3.70 (m, 1 H), 3.21-3.18 (m, 1 H), 2.77-2.76 (m, 1
H), 2.72 (s, 3 H), 2.60-2.41 (m, 4 H), 2.13 (s, 3 H), 2.03-1.99 (m, 1 H), 1.76-1.74 (m, 1 H), 1.57-1.43 (m, 6 H), 1.38 (s, 3 H),
1.21 (d, 3 H, J = 6.8 Hz), 1.17 (s, 3 H), 1.05 (d, 3 H, J = 6.9 Hz). MS (ESI+): 603.3 (M + H) +.
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-17-N-Ethylcarbamoyl-7,11-dihydroxy-8,8,10,12-tetramethyl-3-
[1-methyl-2-(2-methyl- 4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (14a).

OEt
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.98 (s, 1 H), 6.63 (s, 1 H), 5.44-5.32 (m, 1 H), 4.19-4.11 (m, 3 H), 3.91 (br s, 1 H),
3.86 (br s, 1 H), 3.22-3.18 (m, 1 H), 2.71 (s, 3 H), 2.58-2.39 (m, 4 H), 2.24-2.20 (m, 1 H), 2.11 (s, 3 H), 1.90-1.76 (m, 3 H),
1.55-1.41 (m, 3 H), 1.37 (s, 3 H), 1.30-1.27 (m, 3 H), 1.18 (d, 3 H, J = 6.8 Hz), 1.13 (s, 3 H), 1.01 (d, 3H, J = 6.9 Hz). MS
(ESI+): 565.3 (M + H) +.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-Phenylcarbamoyl-7,11-dihydroxy-8,8,10,12-tetramethyl-3-
[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (14b).
OPh
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 7.39-7.34 (m, 2 H), 7.24-7.20 (m, 1 H), 7.11-7.08 (m, 2 H), 6.93 (s, 1 H), 6.61 (s, 1
H), 5.44 (d, 1 H, J = 8.6 Hz), 4.19-4.12 (m, 1 H), 3.87-3.83 (m, 2 H), 3.21-3.18 (m, 1 H), 2.75-2.62 (m, 1 H), 2.67 (s, 3 H),
2.62-2.56 (m, 1 H), 2.55 (dd, 1 H, J = 14.4, 10.2 Hz), 2.47 (dd, 1 H, J = 14.4, 3.3 Hz), 2.32-2.27 (m, 1 H), 2.04 (s, 3 H),
1.92-1.84 (m, 4 H), 1.81-1.44 (m, 5 H), 1.37 (s, 3 H), 1.17 (d, 3 H, J = 6.8 Hz), 1.13 (s, 3 H), 1.01 (d, 3 H, J = 6.9 Hz);
HRMS (ESI+) m/z (M+ + H) calcd for C33H44N2O7S: 613.2948, found: 613.2958.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-Methylsulfonyl-7,11-dihydroxy-8,8,10,12-tetramethyl-3-[1-
methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (15a).
SO2
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.98 (s, 1 H), 6.57 (s, 1 H), 5.34 (d, 1 H, J = 8.0 Hz), 4.15-4.12 (m, 1 H), 3.84-3.81
(m, 1 H), 3.78-3.75 (m, 1 H), 3.18-3.10 (m, 1 H), 3.04 (s, 3 H), 2.87-2.83 (m, 1 H), 2.75-2.71 (m, 1 H), 2.70 (s, 3 H), 2.62-
2.55 (m, 1 H), 2.52 (dd, 1 H, J = 14.7, 10.2 Hz), 2.43 (dd, 1 H, J = 14.7, 3.0 Hz), 2.32-2.27 (m, 1 H), 2.05 (s, 3 H), 1.93-1.70
(m, 3 H), 1.58-1.42 (m, 5 H), 1.36 (s, 3 H), 1.18 (d, 3 H, J = 6.8 Hz), 1.15 (s, 3 H), 1.01 (d, 3 H, J = 6.9 Hz). HRMS (ESI+)
m/z (M+ + H) calcd for C27H42N2O7S2: 571.2512, found: 571.2525.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-n-Propylsulfonyl-7,11-dihydroxy-8,8,10,12-tetramethyl-3-
[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (15b).
SO2
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.99 (s, 1 H), 6.57 (s, 1 H), 5.34 (d, 1 H, J = 8.0 Hz), 3.83-3.82 (m, 1 H), 3.63-3.62
(m, 1 H), 3.18-3.15 (m, 1 H), 3.11- 3.07 (m, 2 H), 2.70 (s, 3 H), 2.67-2.51 (m, 1 H), 2.47- 2.43 (m, 1 H), 2.29- 2.25 (m, 1 H),
2.32-2.27 (m, 1 H), 2.10 (s, 3 H), 2.02-1.70 (m, 5 H), 1.55-1.46 (m, 3 H), 1.37 (s, 3 H), 1.21- 1.08 (m, 12 H), 1.03 (d, 3 H, J
= 6.9 Hz). MS (ESI+) 599.3 (M+ + H).

[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-17-(2-thiophenyl)sulfonyl-7,11-dihydroxy-8,8,10,12-
tetramethyl-3-[1-methyl-2-(2-methyl- 4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (15c).
S
SO2
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 7.70 (m, 2 H), 7.15 (dd, 1 H, J = 4.3, 3.9 Hz), 6.99 (s, 1 H), 6.58 (s, 1 H), 5.36-5.34
(m, 1 H), 3.83-3.80 (m, 1 H), 3.70-3.69 (m, 1 H), 3.18-3.16 (m, 1 H), 2.88-2.83 (m, 1 H), 2.76-2.73 (m, 1 H), 2.71 (s, 3 H),
2.59- 2.53 (m, 2 H), 2.46-2.41 (m, 1 H), 2.19-2.07 (m, 1 H), 2.06 (s, 3 H), 1.92-1.89 (m, 1 H), 1.87-1.70 (m, 3 H), 1.49-1.29
(m, 3 H), 1.27 (s, 3 H), 1.18 (d, 3 H, J = 6.8 Hz), 1.16 (s, 3 H), 1.00 (d, 3 H, J = 6.9 Hz). MS (ESI+): 639.2 (M + H) +.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-Sulfonylurea-7,11-dihydroxy-8,8,10,12-tetramethyl-3-[1-
methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (16a).
H2N
SO2
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.89 (s, 1 H), 6.64 (s, 1 H), 5.39 (dd, 1 H, J = 6.9, 2.3 Hz), 5.20 (br s, 2 H, NH2),
4.15-4.09 (m, 1 H), 3.82-3.77 (m, 2 H), 3.18-3.10 (m, 1 H), 2.65-2.62 (m, 1 H), 2.62 (s, 3 H), 2.51 (dd, 1 H, J = 14.6, 10.4
Hz), 2.48-2.43 (m, 1 H), 2.40 (dd, 1 H, J = 14.6, 3.1 Hz), 2.34-2.29 (m, 1 H), 2.06-2.03 (m, 1 H), 1.95 (s, 3 H), 1.93-1.86 (m,
2 H), 1.74-1.64 (m, 1 H), 1.63-1.34 (m, 5 H), 1.33 (s, 3 H), 1.08 (d, 3 H, J = 6.9 Hz), 1.03 (s, 3 H), 0.91 (d, 3 H, J = 7.0 Hz);
LRMS (ESI+): 572.4 (M + H)+.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-Methylsulfonylurea-7,11-dihydroxy-8,8,10,12-tetramethyl-
3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (16b).
H
N
SO2
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.99 (s, 1 H), 6.61 (s, 1 H), 5.37 (dd, 1 H, J = 9.5, 2.3 Hz), 4.65 (dd, 1 H, J = 10.3,
5.0 Hz), 4.16-4.11 (m, 1 H), 3.83 (dd, 1 H, J = 8.7, 4.4 Hz), 3.75 (d, 1 H, J = 6.5 Hz), 3.20-3.18 (m, 1 H), 2.89 (d, 3 H, J =
5.2 Hz), 2.74-2.70 (m, 1 H), 2.71 (s, 3 H), 2.64-2.55 (m, 1 H), 2.56 (dd, 1 H, J = 14.8, 10.2 Hz), 2.46 (dd, 1 H, J = 14.8, 3.2
Hz), 2.29-2.18 (m, 1 H), 2.08 (s, 3 H), 1.92-1.84 (m, 1 H), 1.83-1.75 (m, 2 H), 1.57-1.39 (m, 6 H), 1.38 (s, 3 H), 1.19 (d, 3 H,
J = 6.7 Hz), 1.15 (s, 3 H), 1.02 (d, 3 H, J = 7.0 Hz); HRMS (ESI+) m/z (M+ + H) calcd for C27H43N3O7S2: 586.2621, found:
586.2635.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N,N-Dimethylsulfonylurea-7,11-dihydroxy-8,8,10,12-
tetramethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (16c).

N
O
SO2
N
S
N
OH
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.99 (s, 1 H), 6.57 (s, 1 H), 5.32 (dd, 1 H, J = 10.2, 2.1 Hz), 4.15-4.12 (m, 1 H),
3.83-3.81 (m, 1 H), 3.65-3.62 (m, 1 H), 3.19-3.16 (m, 1 H), 2.88 (s, 6 H), 2.73-2.69 (m, 1 H), 2.71 (s, 3 H), 2.62-2.55 (m, 1
H), 2.54-2.50 (m, 1 H), 2.52 (dd, 1 H, J = 15.0, 10.0 Hz), 2.45 (dd, 1 H, J = 15.0, 3.0 Hz), 2.29-2.25 (m, 1 H), 2.07 (s, 3 H),
1.86-1.79 (m, 2 H), 1.79-1.75 (m, 1 H), 1.56-1.40 (m, 5 H), 1.37 (s, 3 H), 1.21 (d, 3 H, J = 6.9 Hz), 1.17 (s, 3 H), 1.03 (d, 3
H, J = 6.9 Hz); HRMS (ESI+) m/z (M+ + H) calcd for C28H45N3O7S2: 600.2777, found: 600.2761.
[1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-Morpholinosulfonylurea-7,11-dihydroxy-8,8,10,12-
tetramethyl-3-[1-methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (16d).
O
N
SO2
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.92 (s, 1 H), 6.50 (s, 1 H), 5.23 (d, 1 H, J = 9.3 Hz), 4.06-4.04 (m, 1 H), 3.73-3.72
(m, 1 H), 3.70-3.58 (m, 4 H), 3.20-3.17 (m, 4 H), 3.10-3.08 (m, 1 H), 2.66-2.63 (m, 1 H), 2.64 (s, 3 H), 2.64-2.62 (m, 1 H),
2.61-2.57 (m, 1 H), 2.56 (dd, 1 H, J = 14.8, 10.2 Hz), 2.46 (dd, 1 H, J = 14.8, 3.2 Hz), 2.19-2.14 (m, 1 H), 2.02 (s, 3 H),
1.80-1.72 (m, 1 H), 1.72-1.32 (m, 8 H), 1.29 (s, 3 H), 1.12 (d, 3 H, J = 6.7 Hz), 1.08 (s, 3 H), 0.95 (d, 3 H, J = 6.9 Hz);
LRMS (ESI+): 642.4 (M + H)+.
1S-[1R*,3R*(E),7R*,10S,11R*,12R*,16S*]]-17-N-Ethylurido-7,11-dihydroxy-8,8,10,12-tetramethyl-3-[1-
methyl-2-(2-methyl-4-thiazolyl)ethenyl]-4-oxa-17-azabicyclo[14.1.0]heptadecane-5,9-dione (17).
H
N
O
N
S
N
OH
O
O
O
OH
1H NMR (400 MHz, CDCl3) δ 6.97 (s, 1 H), 6.60 (s, 1 H), 5.62-5.59 (m, 2 H), 4.51 (br s, 1 H, OH), 4.23-4.18 (m, 1
H), 3.86-3.83 (m, 1 H), 3.23-3.20 (m, 3 H), 2.70 (s, 3 H), 2.53-2.42 (m, 2 H), 2.40-2.36 (m, 2 H), 2.09 (s, 3 H), 1.98-1.92 (m,
2 H), 1.82-1.77 (m, 1 H), 1.61-1.36 (m, 7 H), 1.35 (s, 3 H), 1.15 (d, 3 H, J = 7.0 Hz), 1.11 (t, 3 H, J = 7.4 Hz), 1.08 (s, 3 H),
0.97 (d, 3 H, J = 6.9 Hz); HRMS (ESI+) m/z (M+ + H) calcd for C29H45N3O6S: 564.3107, found: 564.3367.