Synthesis and biological activity of analogs of CPZEN-45, a novel antituberculosis drug
as an addition salt of triethylamine. This solid was used in
the next reaction without further purification.
temperature. The reaction solution was concentrated, and to
the resulting syrup was added diethyl ether. The precipitate
obtained was thoroughly washed with diethyl ether to give
5 bis-trifluoroacetate (209 mg) as a colorless solid. The solid
was dissolved in 1-BuOH and the solution was washed
successively with 5% aq sodium hydrogen carbonate and
water. Concentration gave a solid, which was chromato-
graphed (upper layer of 2:1:1 EtOAc−1-PrOH−H2O) to
give 5 trifluoroacetate (114 mg, 59%) as a colorless solid;
[α]D20 + 17° (c 0.5, MeOH); HRESI-MS: m/z calcd for
C32H47N6O12 (M+H)+ 707.3246, found 707.3249; ESI-MS:
m/z 819.3 (M+CF3COOH−H)−; 1H NMR (600 MHz,
DMSO-d6) δ 0.88 (3H, t, J = 7 Hz, CH3CH2CH2CH2−),
1.26 (2H, m, CH3CH2CH2CH2−), 1.46 (2H, m, CH3
CH2CH2CH2−), 2.38 (3H, s, CH3N-5‴), 2.43 (2H, m,
CH3CH2CH2CH2−), 3.00 (1H, slightly br dd, J = ~2 and
14 Hz, H-4‴a), 3.05 (1H, slightly br dd, J = ~2 and ~13 Hz,
H-5″a), 3.14 (1H, slightly br d, J = ~13 Hz, H-5″b), 3.15
(3H, s, CH3N-8‴), 3.22 (1H, slightly br d, J = 14 Hz, H-4‴
b), 3.58 (1H, d, J = 10 Hz, H-6‴), 3.66 (1H, ddd, J = 4.5,
6.5 and 9 Hz, H-3′), 3.73 (1H, t, J = 4.5 Hz, H-2′), 3.76
(1H, t, J = 4 Hz, H-2″), 3.98−4.04 (2H, m, H-3″ and H-4″),
4.09 (1H, d, J = 10 Hz, H-5′), 4.19 (1H, d, J = 9 Hz, H-4′),
4.37 (1H, m, H-3‴), 4.42 (1H, d, J = 5 Hz, H-2‴), 4.83 (1H,
s, H-1′), 4.96 (1H, s, H-1″), 5.15 (1H, d, J = 6.5 Hz, HO-
3′), 5.22 (1H, d, J = 5.5 Hz, HO-3″), 5.24 (2H, d, J = 4 Hz,
HO-2″ and HO-3‴), 5.44 (1H, d, J = 8 Hz, H-5), 5.52 (1H,
d, J = 4 Hz, HO-2′), 6.85 and 7.38 [each 2H, d, J = 8.5 Hz,
CH3(CH2)3C6H4NH], 7.42 (1H, d, J = 8 Hz, H-6), 8.02 (3H,
Compound 10 as an addition salt of triethylamine; ESI-
MS m/z calcd for C27H41N5NaO15 (M+Na)+ 698.3, found
1
698.3; H NMR (600 MHz, DMSO-d6) δ 1.01 [9H, t, J =
7 Hz, (CH3CH2)3N], 1.34 [9H, s, (CH3)3C–O], 2.31 (3H, s,
CH3N-5‴), 2.89 (1H, d, J = 15 Hz, H-4‴a), 2.98 (3H, s,
CH3N-8‴), 3.25 (1H, d, J = 15 Hz, H-4‴b), 3.66 (1H, d,
J = 10 Hz, H-6‴), 4.94 (1H, s, H-1″), 5.55 (1H, d, J = 8 Hz,
H-5), 5.59 (1H, d, J = 2.5 Hz, H-1′), 7.02 (1H, d, J =
8.5 Hz, HN-5″), 7.92 (1H, d, J = 8 Hz, H-6), 11.32 (1H, br
s, HN-3). Anal Calc for C27H41N5O15 Et3N 2H2O: C 48.76,
H 7.44, N 10.34. Found: C 48.55, H 7.58, N 10.05.
5″-N-t-Butoxycarbonylcaprazol 4-butylanilide (11)
To a solution of 10 (an addition salt of triethylamine, 2.28 g,
2.80 mmol) in 2-propanol−water (10:1, 44 mL) were added
4-butylaniline (480 mg, 3.22 mmol) and 4-(4,6-dimethoxy-
1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-
MM) (1.06 g, 3.83 mmol) and the mixture was stirred at
room temperature. The same amounts of 4-butylaniline and
DMT-MM were added after 5 and 15 h, and the mixture
was further stirred for 2 days. Concentration gave a syrup,
which was extracted with 5% aq potassium hydrogen sul-
fate (150 mL) and EtOAc (50 mL). The aqueous solution
was washed with EtOAc and neutralized with sodium
hydrogen carbonate (8.18 g). The precipitate obtained was
extracted with EtOAc and the organic solution was washed
with saturated aq sodium chloride, dried (Na2SO4), and
concentrated. The residue was purified by silica gel column
chromatography (6:1 CHCl3−MeOH) to give 11 (1.03 g,
44%) as a colorless solid; [α]D20 − 11° (c 0.5, MeOH); ESI-
MS: m/z calcd for C37H54N6NaO14 (M+Na)+ 829.4, found
br s, NH3+), 9.48 (1H, s, HN-1‴), 11.16 (1H, s, HN-3); 13
C
NMR (150.9 MHz, DMSO-d6) δ 13.7 (CH3CH2CH2CH2−),
21.7 (CH3CH2CH2CH2−), 32.7 (CH3CH2CH2CH2−), 34.0
(CH3CH2CH2CH2−), 36.3 (CH3N-5‴), 38.8 (CH3N-8‴),
39.6 (C-5″), 58.2 (C-4‴), 63.0 (C-6‴), 66.5 (C-2‴), 67.3 (C-
3‴), 68.4 (C-3′), 69.6 (C-3″), 74.2 (C-2″), 74.4 (C-2′), 74.8
(C-5′), 78.0 (C-4″), 81.4 (C-4′), 89.6 (C-1′), 99.9 (C-5),
109.6 (C-1″), 117.3 (CF3, q, JC, F = 302 Hz), 120.4, 127.2,
135.6, and 137.6 (aromatic), 138.9 (C-6), 149.5 (C-2),
157.6 (CF3COOH, q, JC, F = 30 Hz), 163.2 (C-4), 167.2 (C-
1‴), 169.8 (C-7‴). Anal Calc for C32H46N6O12 CF3COOH
3H2O: C 46.68, H 6.11, N 9.61. Found: C 46.56, H 6.07,
N 9.39.
1
829.3; H NMR (600 MHz, DMSO-d6) δ 0.88 [3H, t, J =
7 Hz, CH3(CH2)3C6H4NH], 1.27 (2H, m, CH3CH2CH2
CH2−), 1.37 [9H, s, (CH3)3C–O], 1.46 (2H, m,
CH3CH2CH2CH2−), 2.37 (3H, s, CH3N-5‴), 2.97 (1H, d,
J = 15 Hz, H-4‴a), 3.13 (3H, s, CH3N-8‴), 3.43 (1H, d,
J = 15 Hz, H-4‴b), 3.65 (1H, d, J = 10 Hz, H-6‴), 4.08
(1H, d, J = 10 Hz, H-5′), 4.18 (1H, d, J = 8 Hz, H-4′), 4.37
(2H, s, H-2‴ and H-3‴), 4.87 (1H, s, H-1′), 4.94 (1H, s,
H-1″), 5.35 (1H, d, J = 8 Hz, H-5), 6.85 and 7.35 [each 2H,
d, J = 8.5 Hz, CH3(CH2)3C6H4NH], 6.92 (1H, slightly br d,
J = 8.5 Hz, HN-5″), 7.58 (1H, d, J = 8 Hz, H-6), 9.38
(1H, s, HN-1‴), 11.14 (1H, s, HN-3). Anal Calc for
C37H54N6O14 2H2O: C 52.72, H 6.94, N 9.97. Found: C
52.95, H 7.24, N 9.99.
Construction of B. subtilis 168 derivatives
overexpressing tagO, mraY and murG
The Bacillus subtilis-Escherichia coli shuttle vector
pHYermA was constructed using pHY300PLK (Takara bio
Inc. Shiga, Japan) [17], as a source of replication origins,
and erythromycin-resistant ermA gene from a Staphylo-
coccus aureus strain. Fragments of tagO and mraY genes of
B. subtilis were cloned into pHYermA, and the resulting
plasmids were designated as pHYermA-tagO and
Caprazol 4-butylanilide (5)
A solution of 11 (187 mg, 0.222 mmol) in trifluoroacetic
acid−methanol (4:1, 5 mL) was kept for 2 h at room