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Synthetic and Immunological Studies on Clustered Modes of Mucin-Related Tn and TF O-Linked Antigens: The Preparation of a Glycopeptide-Based Vaccine for Clinical Trials against Prostate Cancer^†

Supporting Info

Scott D. Kuduk,^‡ Jacob B. Schwarz,^‡ Xiao-Tao Chen,^§ Peter W. Glunz,^‡ Dalibor Sames,^‡^§ Govindaswami Ragupathi, Philip O. Livingston, and Samuel J. Danishefsky*^‡^§

Contribution from the Laboratory for Bioorganic Chemistry and Laboratory for Tumor Vaccinology, Sloan-Kettering Institute For Cancer Research, 1275 York Avenue, New York, New York 10021, and Department of Chemistry, Columbia University, New York, New York 10027

J. Am. Chem. Soc., 1998, 120 (48), pp 12474–12485

DOI: 10.1021/ja9825128

Publication Date (Web): November 20, 1998

†

Abbreviations: BSA, bovine serum albumin; DAST, diethylaminosulfurtrifluoride; DIEA, diisopropylethylamine; ELISA, enzyme-linked immunosorbent assay; FMOC, fluorenylmethoxycarbonyl; GalNAc, N-acetylgalactosamine; IIDQ, 2-isobutoxy-1-isobutoxycarbonyl-1,2-dihryoquinoline; HATU, O-(7-azabenzotriazol-1-yl)-N,N,N‘,N‘-tetramethyluronium hexafluorophosphate; KLH, keyhole limpet hemacyanin; pam, palmitoylcysteine; MBS, m-maleimimidobenzoyl-N-hydroxysuccinimide ester; NHS, N-hydroxysuccinimide; SAMA−OPfp, S-acetylthioglycolic pentafluorophenyl ester; TFA, trifluoroacetic acid; Tn(c), Tn cluster.

‡

Laboratory for Bioorganic Chemistry, Sloan-Kettering Institute For Cancer Research.

Department of Chemistry, Columbia University.

⊥

Laboratory for Tumor Vaccinology, Sloan-Kettering Institute For Cancer Research.

In papers with more than one author, the asterisk indicates the name of the author to whom inquiries about the paper should be addressed.

Abstract

The syntheses of two tumor-associated carbohydrate antigens, Tn and TF, have been achieved using glycal assembly and cassette methodologies. These synthetic antigens were subsequently clustered (c) and immunoconjugated to a carrier protein (KLH or BSA) or a synthetic lipopeptide (pam) for immunological study. Three Tn conjugates were used to vaccinate groups of mice, and all preparations proved to be immunogenic. The Tn(c) covalently linked to KLH (27−KLH) plus the adjuvant QS-21 was the optimal vaccine, inducing high median IgM and IgG titers against Tn(c) by ELISA. These antibodies were strongly reactive with the Tn(c) positive human colon cancer cell line LS-C but not the Tn(c) negative colon cancer cell line LS-B by FACS. The antibodies' reactivities with natural antigens were inhibited with synthetic Tn(c) but not with structurally unrelated compounds. On the basis of these results, vaccines containing 27−KLH and 30−pam plus QS-21 are being tested in patients with prostate cancer.

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