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Synlett 2003(11): 1623-1626
DOI: 10.1055/s-2003-41009
LETTER
© Georg ThiemeVerlag Stuttgart ˙ New York

Synthesis of a Tumor-Associated2,3-Sialyl-T Glycododecapeptide Antigen from the Tandem Repeat Regionof the Mucin MUC1

Sebastian Dziadek, Horst Kunz*
Institut für Organische Chemie, Johannes Gutenberg-Universität Mainz, Duesbergweg 10-14, 55128 Mainz, Germany
Fax: +49(6131)3924786; e-Mail: hokunz@uni-mainz.de;
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Publication History

Received 18 June 2003
Publication Date:
11 August 2003 (online)

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Abstract

As a cell surface antigen for the development of selective antitumorvaccines, a tumor-associated glycododecapeptide from epithelialMUC1 carrying the 2,3-sialyl-T antigen was synthesized. The 2,3-STbuilding block was assembled from a TN-threonine conjugateby stepwise saccharide chain extension and utilized for the solid-phaseglycopeptide synthesis on the fluoride-labile PTMSEL linker.

Key words

2,3-sialyl-T antigen - glycopeptides - MUC1 - regio­selective sialylation - solid-phasesynthesis

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9: [α]D 27 = 32.01(c = 1, CHCl3); Rt = 30.07min [Phenomenex Luna C18 (2), grad.: CH3CN/H2O(60:40) → (90:10), 40min]; 1HNMR (400 MHz, CDCl3, COSY, HMQC): δ (ppm) = 7.74(d, 2 H, H4-, H5-Fmoc, J H4,H3 = J H5,H6 = 7.84 Hz),7.58 (dd, 2 H, H1-, H8-Fmoc, J H1,H2 = J H8,H7 = 7.80 Hz),7.49 (d, 2 H, Har-Bzn, J Ha,Hb = 5.84),7.42-7.35 (m, 2 H, H3-, H6-Fmoc), 7.35-7.24 (m,15 H, H2-, H7-Fmoc, Har-Bzn (3 H), Har-Bn(10 H)), 6.59 (d, 1 H, NH-GalNAc, J NH,H2 = 9.40Hz), 6.12 (d, 1 H, NH-T, J NH,T α = 9.76 Hz),5.45 (s, 1 H, CH-Bzn), 5.40 (t, 1 H, H8′′, J H8 ′′ ,H9 ′′ = 6.64 Hz),5.28-5.13 (m, 2 H, H7′′, NH-NeuNAc),5.16 (s, 2 H, CH2-Bn), 4.98 (d, 1 H, H1, J H1,H2 = 3.52 Hz),4.92-4.81 (m, 1 H, H4′′), 4.74-4.63(m, 1 H, H2), 4.55 (s, 2 H, CH2-Bn), 4.52-4.03(m, 10 H, H9a′′{4.46},CH2-Fmoc {4.39, 4.34}, Tβ {4.33},H4 {4.26}, Tα {4.25},H9-Fmoc {4.19}, H1′ {4.15},H6a {4.13}, H5′′ {4.06}),3.99-3.84 (m, 4H, H6′′ {3.93},H9b′′ {3.92}, H3′ {3.90},H6b {3.87}), 3.72-3.55 (m,4 H, H3 {3.67}, H2′ {3.61},H6a′ {3.59}, H5 {3.57}), 3.54-3.39(m, 2 H, H6b′{3.87}, H5′{3.41}),3.24 (s, 1 H, H4′), 2,78 (sb, 1 H, OH), 2.71(dd, 1 H, H3eq′′, JH3eq ′′ ,H3ax ′′ = 12.72Hz, JH3eq ′′ ,H4 ′′ = 4.28Hz), 2.36 (s, 1 H, OH), 2.03-2.00 (m, 1 H, H3ax′′),2.09, 2.07, 2.02, 2.00, 1.92, 1.82 (5*s, 18H, 6* CH3-Ac),1.42 (s, 9 H, CH3-t-Bu), 1.26(d, 3 H, Tγ, J T γ ,T β = 5.92Hz); 13C NMR (100.6 MHz, CDCl3,BB, HMQC): δ = 171.53, 170.91, 170.82, 170.24,170.09, 169.76, 167.77 (C=O), 156.83 (C=O-urethane),143.71 (C1a-, C8a-Fmoc), 141.29, 141.15 (C4a-, C5a-Fmoc), 138.07,137.51 (Cq-Bn), 134.21 (Cq-Bn), 128.97, 128.84, 128.78,128.43, 127.51 (Car-Bn), 128.78, 128.04, 126.44 (Car-Bzn),127.79 (C3-, C6-Fmoc), 127.09, 126.97 (C2-, C7-Fmoc), 125.17, 124.94(C1-, C8-Fmoc), 120.03 (C4-, C5-Fmoc), 106.55 (C1′), 100.96(CH-Bzn), 100.45 (C1), 97.47 (C2′′), 83.13 (Cq-t-Bu), 78.15 (C3), 75.94, 75.15 (Tβ,C4, C3′), 73.60 (CH2-Bn), 73.43, 72.92 (C6′′,C5′),69.72 (C8′′, C6′), 69.07 (C6), 68.61(C4′′), 68.22, 68.11 (CH2-Bn, C2′,C4′,C7′′),67.25 (CH2-Fmoc), 63.64 (C9′′), 63.52(C5), 59.24 (Tα), 49.08 (C5′′),47.79 (C2), 47.09 (C9-Fmoc), 37.42 (C3′′), 28.06 (CH3-t-Bu), 23.16, 23.07(CH3-NHAc), 21.33, 21.21, 21.01, 20.78, 20.64 (CH3-OAc),19.47 (Tγ), HR-ESI-TOF-MS (positive ion mode):calcd. for C77H91N3O27Na: 1512.5737,found: 1512.5729 [M + Na]+.

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12: [α]D 27 = 24.75(c = 1, CHCl3), HR-ESI-TOF-MS (positive ionmode): calcd. for C74H86N3O31Na2:1558.5040, found: 1558.5024 [M - H + 2 × Na]+.

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Perkin Elmer ABI 433A (Applied Biosystems).

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16: [α]D 26 = -40.68(c = 0.86, H2O), Rt = 14.52min [Phenomenex Jupiter C18, grad.: CH3CN/H2O + 0.1% TFA (5:95) → (30:70),40min]; 1H NMR (400 MHz, D2O,COSY, TOCSY, HMQC): δ (ppm) = 4.92 (d, 1 H, H1, J H1,H2 = 3.72 Hz),4.69 (t, 1 H, Dα, JD α ,D β = 6.36Hz), 4.65-4.58 (m, 2 H, TST α {4.63},Rα {4.62}), 4.57-4.49(m, 1 H, A1 α), 4.49-4.41 (m,3 H, H1′ {4.47}, Sα {4.45},A2 α {4.44}), 4.40-4.34(m, 3 H, 3 × Pα), 4.33-4.26(m, 3 H, TST β {4.31},Vα {4.29}, T1 α {4.28}),4.22-4.14 (m, 3 H, H4 {4.21}, H2 {4.19},T1 β {4.18}), 4.08-3.95(m, 3 H, H3′ {4.04}, H7′′ {4.03},H3 {4.00}), 3.94-3.87 (m, 3 H, Gα {3.91},H4′{3.90}), 3.86-3.52 (m, 20H, H5′, H8′′ {3.83},H6a {3.80}, Sβ {3.76},3 × Pa δ {3.73},H5′′ {3.71}, H9a′′,H9b′′, H4′′ {3.67},3 × Pa α {3.63},H6b {3.62}, H6′′,H5, H6a′, H6b′), 3.48 (dd,1 H, H2′, J H1 ′ ,H2 ′ = 7.92Hz), 3.21-3.14 (m, 2 H, Rδ), 2.99-2.83(m, 2 H, Dβ), 2.71 (dd, 1 H, H3eq′′, J H3eq ′′ ,H3ax ′′ = 12.20Hz, J H3eq ′′ ,H4 ′′ = 4.40Hz), 2.35-2.19 (m, 3 H, 3 × Pa β),2.10-1.94 (m, 8 H, Vβ {2.03},3 × Pγ {2.01}, Ra β),2.02, 1.99, 1.97 (3 × s, 9 H, 3 × CH3-NHAc),1.93-1.77 (m, 4 H, 3 × Pb β {1.88}, H3ax′′ {1.80}),1.77-1.59 (m, 3 H, Rb β{1.70},Rγ {1.63}), 1.38-1.31(m, 6 H, 2 × Aβ), 1.24 (d, 3 H, TST γ, J T γ ,T β = 6.12 Hz),1.15 (d, 3 H, T1 γ, J T γ ,T β = 6.36Hz), 0.94 (d, 6 H, Vγ); MALDI-TOF-MS (dhb, positiveion mode): calcd. for C76H124N17O37:1867.9, found: 1868.8 [M + H]+,1890.9 ([M + Na]+,calcd.: 1889.9), 1907.1 ([M + K]+,calcd.: 1905.9), 1912.8 ([M - H + 2 × Na]+,calcd.: 1911.9); ESI-MS (positive ion mode): calcd. for [C76H121N17O37Na4]2+: 977.88,found: 977.87 [M - 2 H + 4 Na]2+.