M. Kalpio, et al.
ChemistryandPhysicsofLipids231(2020)104937
et al., 2014). For further details, please see Supplementary material.
2.1.2.18. 3-(5Z,8Z,11Z,14Z,17Z)-Eicosa-5,8,11,14,17-pentaenoyl)-1,2-
dihexadecanoyl-sn-glycerol, (R)-5a. (R)-5a was prepared using General
Procedure C, with (S)-3a (100 mg, 0.17 mmol), EPA (64.0 μL, 0.19
mmol, 1.1 equiv.), EDCI (39.0 mg, 0.20 mmol, 1.2 equiv.) and DMAP
(8.0 mg, 0.07 mmol, 0.4 equiv.) in CH2Cl2 (0.4 mL). Purification via
column chromatography (petroleum ether / EtOAc (5 %) gave (R)-4a
(145 mg, 0.17 mmol, 99 %) as a white waxy solid (with a tint of
yellow);
2.1.2.13. General procedure C: Steglich esterification procedure. To a
solution of diacylglycerol (1.0 equiv), N-(3-dimethylaminopropyl)-N’-
ethylcarbodiimide hydrochloride (EDCI) (1.1–1.2 equiv.) and 4-
(dimethylamino)pyridine (DMAP) (0.2−0.4 equiv.) in CH2Cl2 (2.5
mL / mmol substrate), was added the corresponding unsaturated fatty
acid (1.05–1.2 equiv.). After stirring overnight at room temperature,
the resulting crude mixture was directly purified via column
chromatography.
[α]D25 +0.17 (c 15.1, CHCl3); Rf 0.80 (petroleum ether / EtOAc (10
%)); IR (thin film, νmax / cm−1) 3014, 2917, 2849, 1735; 1H NMR (400
MHz, CDCl3) δH 5.46−5.22 (11H, m, 10 × =CH and CHOCO), 4.29
(2H, dd, J = 12.0 and 4.0 Hz, 2 × CH2OCO), 4.18−4.11 (2H, m, 2 ×
CH2’OCO), 2.90−2.75 (8H, m, 4 × =CCH2C=), 2.37−2.26 (6H, m, 3
× CH2COO), 2.16−2.03 (4H, m, =CCH2CH3 and = CCH2CH2), 1.70
(2H, quin, J =7.5 Hz, =CCH2CH2CH2COO), 1.65−1.55 (4H, m, 2 ×
CH2CH2COO), 1.36−1.18 (48H, m, 24 × CH2), 0.98 (3H, t, J =7.5 Hz,
=CCH2CH3), 0.88 (6H, t, J =7.0 Hz, 2 × CH3); 13C NMR (101 MHz,
CDCl3) δC 173.3, 173.0, 172.9, 132.0, 129.0, 128.8, 128.6, 128.3,
128.2, 128.1, 128.0, 127.9, 127.0, 68.9, 62.2, 62.1, 34.2, 34.1, 33.4,
31.9, 29.7, 29.6, 29.5, 29.4, 29.3, 29.2, 29.1, 29.0, 28.9, 26.5, 25.6,
25.5, 24.9, 24.8, 24.7, 22.7, 20.6, 14.3, 14.1; HRMS (ESI+) calc. for
2.1.2.14. 1,2-Dihexadecanoyl-3-(9Z,12Z,15Z)-octadeca-9,12,15-
trienoyl)-sn-glycerol, (R)-4a. (R)-4a was prepared using General
Procedure C, with (S)-3a (195 mg, 0.34 mmol), ALA (109 μL, 0.36
mmol, 1.05 equiv.), EDCI (80.0 mg, 0.42 mmol, 1.2 equiv.) and DMAP
(9.0 mg, 0.07 mmol, 0.2 equiv.) in CH2Cl2 (0.85 mL). Purification via
column chromatography (petroleum ether / EtOAc (5 %) gave (R)-4a
(273 mg, 0.33 mmol, 97 %) as a white waxy solid (with a tint of
yellow);
[α]D25 +0.02 (c 16.3, CHCl3); Rf 0.74 (petroleum ether / EtOAc (10
%)); IR (thin film, νmax / cm−1) 3012, 2917, 2849, 1732; 1H NMR (400
MHz, CDCl3) δH 5.50−5.25 (7H, m, 6 × =CH and CHOCO), 4.29 (2H,
dd, J = 12.0 and 4.0 Hz, 2 × CH2OCO), 4.14 (2H, dd, J = 12.0 and 6.0
Hz, 2 × CH2’OCO), 2.81 (4H, app. t, J =6.5 Hz, 2 × =CCH2C=),
2.37−2.26 (6H, m, 3 × CH2COO), 2.14−2.01 (4H, m, =CCH2CH3 and
= CCH2CH2), 1.67−1.55 (6H, m, 3 × CH2CH2COO), 1.39−1.18 (56H,
m, 28 × CH2), 0.98 (3H, t, J =7.5 Hz, =CCH2CH3), 0.88 (6H, t, J =6.5
Hz, 2 × CH3); 13C NMR (101 MHz, CDCl3) δC 173.4, 173.3, 173.0,
132.1, 130.4, 128.4, 128.3, 127.9, 127.3, 69.0, 62.3 (2), 34.4, 34.3,
34.2, 32.1, 30.1, 30.0, 29.9, 29.8, 29.7, 29.6, 29.5, 29.4, 29.3, 29.2,
29.1, 29.0, 27.4, 25.8, 25.7, 25.1, 25.0, 24.9, 22.8, 20.7, 14.4, 14.3;
HRMS (ESI+) calc. for C53H96O6Na [M + Na]+ 851.7099, found
851.7094.
C
55H96O6Na [M + Na]+ 875.7099, found 875.7101.
2.1.2.19. 1-(5Z,8Z,11Z,14Z,17Z)-Eicosa-5,8,11,14,17-pentaenoyl)-2,3-
dihexadecanoyl-sn-glycerol, (S)-5a. (S)-5a was prepared using General
Procedure C, and the spectroscopic data was identical to that obtained
for (R)-5a. For further details, please see Supplementary material.
2.1.2.20. 3-(5Z,8Z,11Z,14Z,17Z)-Eicosa-5,8,11,14,17-pentaenoyl)-1,2-
dioctadecanoyl-sn-glycerol, (R)-5b. (R)-5b was prepared using General
Procedure C, with (S)-3b (301 mg, 0.48 mmol), EPA (170 μL, 0.53
mmol, 1.1 equiv.), EDCI (111 mg, 0.58 mmol, 1.2 equiv.) and DMAP
(14.0 mg, 0.12 mmol, 0.2 equiv.) in CH2Cl2 (1.2 mL). Purification via
column chromatography (petroleum ether / EtOAc (5 %) gave (R)-5b
(413 mg, 0.45 mmol, 94 %) as a white waxy solid (with a tint of
yellow);
2.1.2.15. 2,3-Dihexadecanoyl-1-(9Z,12Z,15Z)-octadeca-9,12,15-
trienoyl)-sn-glycerol, (S)-4a. (S)-4a was prepared using General
Procedure C, and the spectroscopic data was identical to that
obtained for (R)-4a. For further details, please see Supplementary
material.
[α]D25 +0.18 (c 12.3, CHCl3); Rf 0.70 (petroleum ether / EtOAc (10
%)); IR (thin film, νmax / cm−1) 3013, 2917, 2849, 1735; 1H NMR (400
MHz, CDCl3) δH 5.45−5.23 (11H, m, 10 × =CH and CHOCO),
4.36−4.22 (2H, m, 2 × CH2OCO), 4.18−4.11 (2H, m, 2 × CH2’OCO),
2.90−2.75 (8H, m, 4 × =CCH2C=), 2.40−2.25 (6H, m, 3 ×
CH2COO), 2.17−2.02 (4H, m, =CCH2CH3 and = CCH2CH2), 1.70 (2H,
quin, J =7.5 Hz, =CCH2CH2CH2COO), 1.65−1.56 (4H, m, 2 ×
CH2CH2COO), 1.38−1.18 (56H, m, 28 × CH2), 0.97 (3H, t, J =7.5 Hz,
=CCH2CH3), 0.88 (6H, t, J =7.0 Hz, 2 × CH3); 13C NMR (101 MHz,
CDCl3) δC 173.4, 173.2, 173.0, 132.2, 129.1, 128.9, 128.7, 128.4,
128.3, 128.2, 128.0, 127.2, 69.0, 62.4, 62.3, 34.4, 34.2, 33.6, 32.1,
29.9, 29.8, 29.7, 29.6, 29.5, 29.4, 29.3, 29.2, 29.1, 29.0, 26.7, 25.8,
25.7, 25.1, 25.0, 24.9, 22.9, 20.7, 14.4, 14.3; HRMS (ESI+) calc. for
2.1.2.16. 1,2-Dioctadecanoyl-3-(9Z,12Z,15Z)-octadeca-9,12,15-trienoyl)-
sn-glycerol, (R)-4b. (R)-4b was prepared using General Procedure C,
with (S)-3b (302 mg, 0.48 mmol), ALA (168 μL, 0.56 mmol, 1.15
equiv.), EDCI (114 mg, 0.60 mmol, 1.2 equiv.) and DMAP (14.0 mg,
0.12 mmol, 0.2 equiv.) in CH2Cl2 (1.2 mL). Purification via column
chromatography (petroleum ether / EtOAc (5 %) gave (R)-4b (411 mg,
0.46 mmol, 96 %) as a white waxy solid (with a tint of yellow);
[α]D25 +0.02 (c 21.7, CHCl3); Rf 0.70 (petroleum ether / EtOAc (10
%)); IR (thin film, νmax / cm−1) 3012, 2917, 2849, 1733; 1H NMR (400
MHz, CDCl3) δH 5.45−5.23 (7H, m, 6 × =CH and CHOCO), 4.29 (2H,
dd, J = 12.0 and 4.0 Hz, 2 × CH2OCO), 4.14 (2H, dd, J = 12.0 and 6.0
Hz, 2 × CH2’OCO), 2.87−2.74 (4H, m, 2 × =CCH2C=), 2.35−2.26
(6H, m, 3 × CH2COO), 2.14−2.01 (4H, m, =CCH2CH3 and =
CCH2CH2), 1.67−1.56 (6H, m, 3 × CH2CH2COO), 1.38−1.18 (64H, m,
32 × CH2), 0.98 (3H, t, J =7.5 Hz, =CCH2CH3), 0.88 (6H, t, J =7.0
Hz, 2 × CH3); 13C NMR (101 MHz, CDCl3) δC 173.5, 173.4, 173.0,
132.1, 130.4, 128.5, 128.4, 127.9, 127.3, 69.0, 62.3 (2), 34.4, 34.3,
34.2, 32.1, 30.1(2), 30.0(2), 29.9, 29.8, 29.7, 29.6, 29.5, 29.4, 29.3,
29.2, 29.1, 29.0, 27.4, 25.8, 25.7, 25.1, 25.0, 24.9, 22.8, 20.7, 14.4,
14.3; HRMS (ESI+) calc. for C57H104O6Na [M + Na]+ 907.7725,
found 907.7717.
C
59H104O6Na [M + Na]+ 931.7725, found 931.7701.
2.1.2.21. 1-(5Z,8Z,11Z,14Z,17Z)-Eicosa-5,8,11,14,17-pentaenoyl)-1,2-
dioctadecanoyl-sn-glycerol, (S)-5b. (S)-5b was prepared using General
Procedure C, and the spectroscopic data was identical to that obtained
for (R)-5b. For further details, please see Supplementary material.
2.1.2.22. 3-(4Z,7Z,10Z,13Z,16Z,19Z)-Docosa-4,7,10,13,16,19-
hexaenoyl)-1,2-dihexadecan-oyl-sn-glycerol,
(R)-6a. (R)-6a
was
prepared using General Procedure C, with (S)-3a (331 mg, 0.58
mmol), DHA (213 μL, 0.61 mmol, 1.05 equiv.), EDCI (122 mg, 0.64
mmol, 1.1 equiv.) and DMAP (14.0 mg, 0.12 mmol, 0.2 equiv.) in
CH2Cl2 (1.5 mL). Purification via column chromatography (petroleum
ether / EtOAc (5 %) gave (R)-6a (512 mg, 0.58 mmol, 99 %) as a white
waxy solid (with a tint of yellow);
2.1.2.17. 2,3-Dioctadecanoyl-1-(9Z,12Z,15Z)-octadeca-9,12,15-trienoyl)-
sn-glycerol, (S)-4b. (S)-4b was prepared using General Procedure C, and
the spectroscopic data was identical to that obtained for (R)-4b. For
further details, please see Supplementary material.
[α]D25 +0.08 (c 13.5, CHCl3); Rf 0.70 (petroleum ether / EtOAc (10
%)); IR (thin film, νmax / cm−1) 3014, 2917, 2849, 1734; 1H NMR (400
4