and 120 mg (0.18 mmol) of saquinavir gave after purification by
chromatography (3 : 7 to 0 : 7 hexane/AcOEt) 160 mg (0.12
mmol, 65%) of Saq-C(O)C2C(O)GlyOleoyl: Rf (3 : 7 hexane/
AcOEt) = 0.4; 1H NMR (CDCl3): 0.84 (t, J 6.4, 6H, H1Ј), 1.25
(s, 40H, H2Ј–7Ј, H12Ј–15Ј), 1.30 (s, 9H, H41), 1.45–1.85 (m,
12H, H31–34, H16Ј), 1.98 (m, 8H, H8Ј, H11Ј), 2.28 (t, J 7.5,
4H, H17Ј), 2.20–3.00 (m, 17H, H13, H19, H27, H29, H30,
H35–37, H22Ј, H23Ј), 4.15, 4.25 (2m, 4H, H19Ј), 4.39 (m, 1H,
H18), 4.82 (m, 1H, H12), 5.15–5.40 (m, 6H, H26, H9Ј, H10Ј,
H20Ј), 5.97 (bs, 1H, H39), 6.39 (bs, 1H, H15), 6.53 (bs, 1H,
H15), 6.91 (m, 1H, H23), 7.00–7.15 (m, 4H, H21, H22, H24,
H25), 7.58 (m, 2H, H3, H17), 7.71 (td, J 1.6, 8.0, 1H, H2), 7.80
(d, J 8.0, 1H, H4), 8.09 (d, J 8.0, 1H, H1), 8.13 (d, J 8.3, 1H,
H6), 8.22 (d, J 8.3, 1H, H7), 9.12 (d, J 7.4, 1H, H11); 13C NMR
(CDCl3): 14.2 (C1Ј), 20.7, 25.8, 26.2, 30.7 (C31–34), 22.7 (C2Ј),
24.9 (C16Ј), 27.3 (C8Ј, C11Ј), 28.8 (C41), 29.1, 29.2, 29.4, 29.6,
29.8 (C4Ј–7Ј, C12Ј–15Ј, C22Ј, C23Ј), 30.9 (C36), 31.9 (C3Ј),
33.2 (C35), 34.1 (C17Ј), 35.0 (C19), 35.8 (C30), 37.5 (C13), 49.7
(C18), 50.9 (C40), 51.7 (C12), 56.6 (C27), 59.5 (C29), 61.9
(C19Ј), 69.8 (C20Ј), 70.6 (C37), 73.8 (C26), 118.7 (C7), 126.5
(C23), 127.6 (C3), 128.1 (C4), 128.5 (C22, C24), 129.1 (C21,
C25), 129.3 (C5), 129.8, 130.1 (C9Ј, C10Ј), 130.2 (C1, C2), 137.2
(C20), 137.3 (C6), 146.6 (C9), 149.1 (C8), 164.5 (C10), 170.3
(C38), 171.8, 171.9 (C21Ј, C24Ј), 173.3, 173.4, 173.6 (C14, C16,
C18Ј); ESI(ϩ) MS: m/z = 1374.09 (M ϩ H)ϩ in agreement with
the calculated mass for [M] = C81H124N6O12.
(C20), 171.1, 171.6, 172.0, 172.1 (C16Ј), 173.0, 173.2 (C19Ј,
C22Ј), 174.4 (C11); ESI(ϩ) MS: m/z = 1915.17 (M)ϩ and
1916.24 (M ϩ H)ϩ in agreement with the calculated mass for
[M] = C114H187N5O18.
Ind(8)-C(O)C2C(O)GlyOleoyl and Ind-[C(O)C2C(O)-
GlyOleoyl]2. The ester procedure when applied to 252 mg (0.35
mmol) of HOC(O)C2C(O)GlyOleoyl and 180 mg (0.29 mmol)
of indinavir gave after purification by chromatography (1 : 0 to
0 : 1 hexane/AcOEt) 155 mg (0.12 mmol, 40%) of Ind(8)-
C(O)C2C(O)GlyOleoyl and 77 mg (0.04 mmol, 13%) of
Ind-[C(O)C2C(O)GlyOleoyl]2.
Ind(8)-C(O)C2C(O)GlyOleoyl. Rf (10 : 1 AcOEt/MeOH) =
1
0.5; H NMR (CDCl3): 0.85 (t, J 6.0, 6H, H1Ј), 1.20 (s, 9H,
H23), 1.25 (s, 24H, H2Ј–7Ј), 1.30 (s, 16H, H12Ј–15Ј), 1.50–1.65
(m, 6H, H13, H16Ј), 1.99 (m, 8H, H8Ј, H11Ј), 2.20–3.20 (m,
22H, H7, H12, H15–19, H30, H17Ј, H22Ј, H23Ј), 3.47 (s, 2H,
H24), 3.80 (m, 1H, H14), 3.95–4.21 (m, 4H, H19Ј), 4.97 (m, 1H,
H20Ј), 5.31 (m, 5H, H8, H9Ј, H10Ј), 5.63 (dd, J 5.1, 9.1, 1H,
H9), 6.45 (d, J 9.1, 1H, H10), 7.12–7.29 (m, 10H, H2–5, H27,
H32–36), 7.58 (d, J 7.8, 1H, H26), 7.65 (bs, 1H, H21), 8.50 (m,
2H, H28, H29); 13C NMR (CDCl3): 14.2 (C1Ј), 22.7 (C2Ј), 24.9
(C16Ј), 27.2 (C8Ј, C11Ј), 29.1, 29.2, 29.4, 29.6, 29.7, 29.8 (C4Ј–
7Ј, C12Ј–15Ј, C22Ј, C23Ј), 29.0 (C23), 31.9 (C3Ј), 34.0 (C17Ј),
37.6 (C7), 38.2 (C13), 39.4 (C30), 46.1 (C12), 48.1 C18), 51.2
(C22), 52.8, 54.8 (C16, C17), 55.2 (C9), 60.3 (C24), 61.6 (C15),
61.7 (C19Ј), 64.4 (C19), 65.9 (C14), 69.7 (C20Ј), 77.4 (C8), 123.5
(C27), 123.7 (C5), 125.1 (C34), 126.3, 127.0 (C3, C4), 128.1
(C2), 128.4 (C33, C35), 129.0 (C32, C36), 129.8, 130.1 (C9Ј,
C10Ј), 132.6 (C25), 136.8 (C26), 139.3, 139.9 (C1, C6), 141.0
(C31), 149.1 (C28), 150.6 (C29), 169.4 (C20), 170.8, 171.7
(C21Ј, C24Ј), 173.2, 173.3 (C18Ј), 175.1 (C11); ESI(ϩ) MS: m/z
= 1316.91 (M ϩ H)ϩ and 1338.91 (M ϩ Na)ϩ in agreement with
the calculated mass for [M] = C79H121N5O11.
Ind(8)-C(O)C2C(O)GlyPalm and Ind-[C(O)C2C(O)-
GlyPalm]2. The ester procedure when applied to 234 mg (0.35
mmol) of HOC(O)C2C(O)GlyPalm and 195 mg (0.32 mmol) of
indinavir gave after purification by chromatography (1 : 0 to
0 : 1 hexane/AcOEt) 157 mg (0.12 mmol, 39%) of Ind(8)-
C(O)C2C(O)GlyPalm and 73 mg (0.04 mmol, 12%) of
Ind-[C(O)C2C(O)GlyPalm]2.
Ind(8)-C(O)C2C(O)GlyPalm. Rf (10 : 1 AcOEt/MeOH) =
0.5; H NMR (CDCl3): 0.85 (t, J 6.4, 6H, H1Ј), 1.23 (s, 48H,
Ind-[C(O)C2C(O)GlyOleoyl]2. Rf (AcOEt) = 0.4; 1H NMR
(CDCl3): 0.84 (t, J 6.4, 12H, H1Ј), 1.20 (s, 9H, H23), 1.25 (s,
48H, H2Ј–7Ј), 1.30 (s, 32H, H12Ј–15Ј), 1.45–1.70 (m, 10H, H13,
H16Ј), 1.90–2.15 (m, 16H, H8Ј, H11Ј), 2.20–3.15 (m, 30H, H7,
H12, H15–19, H30, H17Ј, H22Ј, H23Ј), 3.43 (s, 2H, H24), 3.60–
4.25 (m, 8H, H19Ј), 4.71 (m, 1H, H20Јa), 4.93 (m, 1H, H20Јb),
5.30 (m, 9H, H8, H9Ј, H10Ј), 5.68 (dd, J 5.0, 9.3, 1H, H9), 6.55
(d, J 9.3, 1H, H10), 6.86 (bs, 1H, H21), 7.11–7.33 (m, 10H, H2–
5, H27, H32–36), 7.60 (d, J 7.8, 1H, H26), 8.46 (m, 2H, H28,
H29); 13C NMR (CDCl3): 14.2 (C1Ј), 22.7 (C2Ј), 24.9 (C16Ј),
27.3 (C8Ј, C11Ј), 28.9 (C23), 29.2, 29.3, 29.4, 29.6, 29.8 (C4Ј–7Ј,
C12Ј–15Ј, C22Ј, C23Ј), 31.9 (C3Ј), 34.0 (C17Ј), 35.3 (C13), 37.5
(C7), 39.9 (C30), 45.1 (C12), 50.6 (C18), 50.9 (C22), 52.2, 55.9
(C16, C17), 55.2 (C9), 59.1 (C15), 59.9 (C24), 61.2, 61.4, 61.6
(C19Ј), 67.3 (C19), 69.6 (C20Ј), 70.3 (C14), 76.8 (C8), 123.4
(C27), 124.1 (C5), 124.9 (C34), 126.4, 127.2 (C3, C4), 128.1
(C2), 128.5 (C33, C35), 129.0 (C32, C36), 129.7, 130.1 (C9Ј,
C10Ј), 133.1 (C25), 136.7 (C26), 139.2, 139.3 (C1, C6), 140.5
(C31), 148.8 (C28), 150.4 (C29), 170.5 (C20), 171.1, 171.6,
172.0 (C18Ј), 173.0, 173.2 (C21Ј, C24Ј), 174.4 (C11); ESI(ϩ)
MS: m/z = 2019.45 (M ϩ H)ϩ in agreement with the calculated
mass for [M] = C122H195N5O18.
1
H2Ј–13Ј), 1.30 (s, 9H, H23), 1.45–1.70 (m, 6H, H13, H14Ј),
2.20–3.15 (m, 22H, H7, H12, H15–19, H30, H15Ј, H20Ј, H21Ј),
3.45 (s, 2H, H24), 3.85 (m, 1H, H14), 3.90–4.20 (m, 4H, H17Ј),
4.98 (m, 1H, H18Ј), 5.30 (m, 1H, H8), 5.64 (dd, J 5.0, 9.1, 1H,
H9), 6.43 (d, J 9.1, 1H, H10), 7.12–7.30 (m, 10H, H2–5, H27,
H32–36), 7.60 (d, J 7.8, 1H, H26), 7.68 (bs, 1H, H21), 8.50 (m,
2H, H28, H29); 13C NMR (CDCl3): 14.2 (C1Ј), 22.7 (C2Ј), 24.9
(C14Ј), 29.1, 29.2, 29.3, 29.4, 29.5, 29.7 (C4Ј–13Ј, C20Ј, C21Ј),
28.9 (C23), 32.0 (C3Ј), 34.1 (C15Ј), 37.6 (C7), 38.3 (C13), 39.4
(C30), 46.1 (C12), 48.0 (C18), 51.2 (C22), 52.8, 54.8 (C16, C17),
55.2 (C9), 60.3 (C24), 61.5 (C15), 61.7 (C17Ј), 64.4 (C19), 65.9
(C14), 69.7 (C18Ј), 77.2 (C8), 123.5 (C27), 123.7 (C5), 125.1
(C34), 126.3, 127.0 (C3, C4), 128.1 (C2), 128.4 (C33, C35),
129.1 (C32, C36), 132.6 (C25), 136.8 (C26), 139.4, 139.9 (C1,
C6), 141.0 (C31), 149.2 (C28), 150.6 (C29), 169.4 (C20), 170.8,
171.7 (C19Ј, C22Ј), 173.3, 173.4 (C16Ј), 175.1 (C11); ESI(ϩ)
MS: m/z = 1265.03 (M ϩ H)ϩ and 1287.89 (M ϩ Na)ϩ in
agreement with the calculated mass for [M] = C75H117N5O11.
1
Ind-[C(O)C2C(O)GlyPalm]2. Rf (AcOEt) = 0.4; H NMR
(CDCl3): 0.85 (t, J 6.4, 12H, H1Ј), 1.20 (s, 96H, H2Ј–13Ј), 1.30
(s, 9H, H23), 1.45–1.70 (m, 10H, H13, H14Ј), 2.25–3.10 (m,
30H, H7, H12, H15–19, H30, H15Ј, H20Ј, H21Ј), 3.44 (s, 2H,
H24), 3.65–4.25 (m, 8H, H17Ј), 4.73 (m, 1H, H18Јa), 4.95 (m,
1H, H18Јb), 5.16 (m, 1H, H14), 5.30 (m, 1H, H8), 5.71 (dd,
J 4.8, 9.5, 1H, H9), 6.53 (d, J 9.5, 1H, H10), 6.86 (bs, 1H, H21),
7.13–7.34 (m, 10H, H2–5, H27, H32–36), 7.61 (d, J 7.8, 1H,
H26), 8.47 (m, 2H, H28, H29); 13C NMR (CDCl3): 14.2 (C1Ј),
22.9 (C2Ј), 24.9 (C14Ј), 29.2, 29.4, 29.6, 29.8 (C4Ј–13Ј, C20Ј,
C21Ј), 28.9 (C23), 32.0 (C3Ј), 34.0 (C15Ј), 35.3 (C13), 37.5 (C7),
40.0 (C30), 45.2 (C12), 50.5 (C18), 50.9 (C22), 52.2, 55.9 (C16,
C17), 55.3 (C9), 59.1 (C15), 60.0 (C24), 61.2, 61.4, 61.7 (C17Ј),
67.4 (C19), 69.6 (C18Ј), 70.3 (C14), 76.8 (C8), 123.4 (C27),
124.1 (C5), 124.9 (C34), 126.4, 127.2 (C3, C4), 128.1 (C2), 128.5
(C33, C35), 129.0 (C32, C36), 133.1 (C25), 136.7 (C26), 139.2,
139.3 (C1, C6), 140.5 (C31), 148.9 (C28), 150.4 (C29), 170.5
Hydrolysis kinetics
The hydrolysis experiments were performed by incubating
20 mL of a DMEM/MeOH solution (pH 7.3) of the prodrug
(250 µg mLϪ1) at 37 ЊC under stirring. The MeOH amount
(v/v) of these solutions was 6% for Saq-Phe(2TFA), 6%
for Saq-Leu(2TFA), 5.1% for Saq-C(O)C4Tyr(1TFA) and
Saq-C(O)NC4Tyr(1TFA), 5% for Ind(8)-Val(4TFA), Ind(14)-
Val(4TFA), Ind(8)-Phe(2TFA), Ind(8)-Leu(2TFA), Ind-
[C(O)C4Tyr]2 (1TFA), and Nelf(1)-C(O)C4Tyr(1TFA), 4% for
Nelf(18)-C(O)NC4Tyr(1TFA), 3% for Ind(8)-C(O)C4Tyr-
(1TFA), and 2% for Ind(14)-C(O)NC4Tyr(1TFA). Hydrolysis
was followed by HPLC monitoring of the disappearance of the
prodrug and appearance of the parent drug [this was the case
O r g . B i o m o l . C h e m . , 2 0 0 4 , 2, 3 4 5 – 3 5 7
355