Synthesis of a "memory tripeptide"
Russ.Chem.Bull., Int.Ed., Vol. 59, No. 1, January, 2010
207
1190 (P—O—Alk); 1250 (P=О); 1690 (C=O, amide); 1745
(C=O, acidic); 3280 (NH).
(8 mL), and 2,3ꢀdihydroꢀ1Hꢀindenꢀ1ꢀone (0.4 g, 3 mmol), moꢀ
lecular sieves 4 Å (500 mg), PcAlCl (0.08 g, 0.1 mmol), and
t
Nꢀ[(Diethoxyphosphoryl)phenyl)methyl]ꢀLꢀγꢀglutamylꢀLꢀcysꢀ
teinylglycine (15). Glutathione (13) (0.68 g, 2.2 mmol), molecuꢀ
lar sieves 4 Å (500 mg), tPcAlCl (0.08 g, 0.1 mmol), and triethylꢀ
amine (0.46 mL, 3.3 mmol) were added to a solution of benzalꢀ
dehyde (0.2 mL, 2 mmol) in 2,2,2ꢀtrifluoroethanol (4 mL). The
reaction mixture was heated to 60 °C and stirred for 3 h, and
diethyl phosphite (0.39 mL, 3 mmol) was added. The mixture
was refluxed with stirring for 120 h. Molecular sieves were
filtered off and washed with methanol (3×2 mL), and the filtrate
was concentrated in vacuo. After column chromatography
(CHCl3—MeOH, 15 : 1), an orange oily substance was obꢀ
tained in a yield of 0.46 g (40%), Rf 0.37 (CHCl3 : MeOH, 10 : 1).
Found (%): C, 47.65; H, 6.27; N, 7.74. C21H32N3O9PS. Calcuꢀ
lated (%): C, 47.27; H, 6.05; N, 7.88. 1Н NMR, δ: 1.20, 1.25
(both t, 6 H, 2 CH3 (Et), J = 7.07 Hz); 1.81—2.04, 2.23—2.50
DIEA (0.35 mL, 2 mmol) were added. The reaction mixture was
stirred for 3 h at 30 °C, diethyl phosphite (0.2 mL, 1.5 mmol) was
added, and the mixture was stirred for 1 day at 30 °C. Then more
diethyl phosphite (0.2 mL, 1.5 mmol) was added. The reaction
was monitored by TLC, this lasted for 120 h. The molecular
sieves were filtered off and washed with methanol (3×2 mL), and
the filtrate was concentrated in vacuo. After column chromatogꢀ
raphy (CHCl3—MeOH, 20 : 1), an orange oily substance was
obtained in a yield of 0.59 g (60%). 1Н NMR, δ: 1.18—1.35 (m, 6 H,
2 CH3); 1.58 (br.m, 4 H, CHCH2CH2CH2, 2 Arg); 1.73, 1.91
(both br.m, 4 H, CHCH2CH2CH2, 2 Arg); 1.98, 2.10 (both m,
2 H, CHCH2CH2, Glu); 2.30—2.60 (m, 4 H, CHCH2CH2, Glu,
C(2)H2, cycl.); 2.87—3.06 (m, 2 H, 3CH2, cycl.); 3.12—3.33
(br.m, 4 H, CHCH2CH2CH2, 2 Arg); 3.57, 3.81 (both m, H,
CH, Arg); 4.01, 4.62 (both m, 6 H, CH, Arg, CH, Glu, 2 OCH2);
5.09 (m, 4 H, 2 CH2Ph); 7.18—7.76 (m, 14 H, arom.). 31P NMR,
δ: 25.17, 25.62. 13C NMR, δ: 15.97—16.21 (Me, POEt); 24.37,
24.48 (C(4), 2 Arg); 26.58 (C(3), Glu); 28.83 (C(4), Glu); 30.08
(C(3), 2 Arg); 32.01 (3CH2, cycl.); 33.28 (2CH2, cycl.); 40.67
(C(5), 2 Arg); 51.86, 52.53 (C(2), Glu); 52.96, 54.49, 55.50,
55.84 (C(2), 2 Arg); 63.21, 64.04 (OCH2); 66.81, 67.62 (CH2Ph);
68.06, 68.40 (both d, C(1), 1JC,P = 158.8 Hz, 1JC,P = 159.5 Hz);
125.14, 126.48, 126.69, 128.19, 128.38, 128.63, 129.12, 134.93,
135.46, 139.33, 144.81 (C arom.); 159.17 (br.s, C=NH, 2 Arg);
171.40, 173.24 (NHC=O, Arg, Glu); 173.50, 176.34 (C(O)OBzl,
Arg, Glu). IR, ν/cm–1: 970, 1190 (P—O—Alk); 1250 (br, N=O,
NO2, P=О); 1600, 1630 (br, N=O, NO2, C=N, amide); 1730
(C=O); 3300 (NH). MS (MALDI—TOF), m/z: 981 [М]+, 844
[M – P(O)(OEt)2]+.
3
(both m, 4 H, CH2CH2, Glu); 3.45—3.87 (m, 3 H, CH2, Cys,
2CH, Glu); 3.93—4.25 (m, 6 H, 2 OCH2, CH2, Gly); 4.31—4.37
(m, H, 2CH, Cys); 5.01 (d, αꢀCH, 2JHP = 11.1 Hz); 7.28—7.39,
7.49 (m, 5 H, arom.). 31P NMR, δ: 21.54.
Benzyl Nꢀ(1ꢀ(diethoxyphosphoryl)ꢀ2,3ꢀdihydroꢀ1Hꢀindenꢀ1ꢀ
yl)ꢀ(γꢀbenzyl)ꢀαꢀLꢀglutamylꢀNωꢀnitroꢀLꢀargininate (16). 2,3ꢀDihyꢀ
droꢀ1Hꢀindenꢀ1ꢀone (0.4 g, 3 mmol), molecular sieves 4 Å
(500 mg), PctAlCl (0.08 g, 0.1 mmol), and diisopropylethylamine
(0.35 mL, 2 mmol) were added to a solution of dipeptide 3b
(1.3 g, 2 mmol) in dichloromethane (7 mL). The reaction mixꢀ
ture was stirred for 3 h at 30 °C, diethyl phosphite (0.39 mL,
3 mmol) was added, and the mixture was stirred for 1 day at 30 °C.
Then more diethyl phosphite (2 mmol) was added. The duration
of the reaction was 72 h (TLC). Molecular sieves were filtered
off and washed with methanol (3×2 mL), and the filtrate
was concentrated in vacuo. After column chromatography
(CHCl3—MeOH, 20 : 1), an orange oily substance was obtained
in a yield of 1.09 g (60%). 1Н NMR, δ: 1.30 (t, 6 H, 2 CH3,
J = 7.07 Hz); 1.57 (br.m, 2 H, CHCH2CH2CH2, Arg); 1.67, 1.83
(both br.m, 2 H, CHCH2CH2CH2, Arg); 1.86, 2.01 (both m,
2 H, CHCH2CH2, Glu); 2.37—2.66 (m, 4 H, CHCH2CH2,
Glu, CH2 cycl.); 3.24 (m, 2 H, CH2 cycl.); 3.41 (br.m, 2 H,
CHCH2CH2CH2, Arg); 3.58 (m, 1 H, CH, Arg); 4.06—4.14
(m, 4 H, 2 OCH2); 4.56 (m, 1 H, CH, Glu); 5.13 (m, 4 H,
CH2Ph); 7.32 (m, 14 H, arom.). 31P NMR, δ: 30.35. 13C NMR,
δ: 16.56 (CH3); 24.39 (C(4), Arg); 29.63 (C(3), Glu); 30.23 (C(4),
Glu); 30.76 (C(3), Arg); 31.41 (C(3)H2, cycl.); 33.18 (C(2)H2,
cycl.); 40.32 (C(5), Arg); 50.71 (C(2), Glu); 53.18 (d, C(1), cycl.,
1JC,P =149.3 Hz); 56.12 (C(2), Arg); 62.40, 62.62 (both d, OCH2,
2JC,P = 7.3 Hz); 66.54, 67.45 (CH2Ph); 125.6, 126.71, 127.05,
127.33, 128.24, 128.51, 128.55, 128.66, 135.00, 135.74, 138.46,
142.10 (C arom.); 159.45 (C=NH, Arg); 171.54 (NHC=O,
Glu); 173.11, 176.18 (C(O)OBzl, Arg, Glu). IR, ν/cm–1: 970,
1190 (P—O—Alk); 1250 br (N=O, NO2, P=О); 1600, 1630 br
(N=O, NO2, C=N, amide); 1730 (C=O); 3300 (NH). MS
(MALDI—TOF), m/z: 780 [М]+, 643 [M – P(O)(OEt)2]+.
Benzyl [Nαꢀ(1ꢀ(diethoxyphosphoryl)ꢀ2,3ꢀdihydroꢀ1Hꢀindenꢀ
1ꢀyl)ꢀNωꢀnitroꢀLꢀarginyl]ꢀ(γꢀbenzyl) αꢀLꢀglutamylꢀNωꢀnitroꢀLꢀ
argininate (17). Tripeptide 4a (0.83 g, 1 mmol) was dissolved in
a mixture of trifluoroacetic acid (15 mL) and dichloromethane
(15 mL). The solution was stirred for ~4 h until the starting
compound disappeared from the reaction mixture (TLC). The
solvents were evaporated in vacuo to a constant weight. The
obtained glassy substance was dissolved in dichloromethane
Benzyl [Nαꢀ(5ꢀbenzyloxycarbonylꢀ1ꢀ(diethoxyphosphoryl)ꢀ
2,3ꢀdihydroꢀ1Hꢀindenꢀ1ꢀyl)ꢀNωꢀnitroꢀLꢀarginyl]ꢀ(γꢀbenzyl) αꢀLꢀ
glutamylꢀNωꢀnitroꢀLꢀargininate (18) was synthesized using benꢀ
zylꢀ1ꢀoxoꢀ2,3ꢀdihydroꢀ1Hꢀindeneꢀ5ꢀcarboxylate as the carbonꢀ
yl component using the method of synthesis of αꢀaminophosꢀ
phonate 17. An orange oily substance was obtained in a yield
of 0.68 g (60%). 1Н NMR, δ: 1.09—1.35 (m, 6 H, 2 CH3);
1.46—1.82 (br.m, 8 H, CHCH2CH2CH2); 1.86—2.12 (m, 2 H,
CHCH2CH2, Glu); 2.34—2.57 (m, 4 H, CHCH2CH2, Glu,
C(2)H2, cycl.); 2.87—3.00 (m, 2 H, C(3)H2, cycl.); 3.10—3.37
(br.m, 4 H, CHCH2CH2CH2, 2 Arg); 3.56, 3.73 (both m, 1 H,
CH, Arg); 4.00—4.65 (m, 6 H, CH, Arg, CH, Glu, 2 OCH2);
5.09—5.17 (m, 6 H, 2 CH2Ph); 7.10—7.78 (m, 19 H, arom.).
31P NMR, δ: 27.66, 27.77. 13C NMR, δ: 15.96—16.20 (Me,
POEt); 24.26, 24.49 (C(4), 2 Arg); 26.62 (C(3), Glu); 28.79 (C(4),
Glu); 29.66 (C(3)H2, cycl.); 30.06 (C(3), 2 Arg); 32.03 (C(2)H2,
cycl.); 40.67 (C(5), 2 Arg); 51.66, 51.77 (C(2), Glu); 52.95, 54.49,
55.50, 55.85 (C(2), 2 Arg); 63.20, 64.00 (OCH2); 66.52, 66.81,
1
67.62 (CH2Ph); 68.70, 69.10 (both d, C(1), JC,P = 157.4 Hz,
1JC,P = 158.5 Hz); 124.23, 125.64, 128.18, 128.37, 128.63, 129.07,
129.73, 134.91, 135.47, 136.41, 144.57, 150.17 (C arom.); 159.18,
(br.s, C=NH, 2 Arg); 171.41, 171.84 (NHC=O, Arg, Glu);
173.22, 173.49, 176.32 (C(O)OBzl, Arg, Glu). IR, ν/cm–1: 970,
1190 (P—O—Alk); 1250 (br, N=O, NO2, P=О); 1600, 1630
(br, N=O, NO2, C=N, amide); 1730 (C=O); 3300 (NH). MS
(MALDI—TOF), m/z: 1115 [М]+, 978 [M – P(O)(OEt)2]+.
{Nαꢀ[1ꢀ(Diethoxyphosphoryl)ꢀ2,3ꢀdihydroꢀ1Hꢀindenꢀ1ꢀyl]ꢀ
Lꢀarginyl}ꢀαꢀLꢀglutamylꢀLꢀarginine (19). A flow of hydrogen was
passed through a solution of αꢀaminophosphonate 17 (0.2 g) in
a mixture of acetic acid (8 mL) and water (2 mL) (300 mL h–1
)