title: Disulfide-linked antibody-maytansinoid conjugates: optimization of in vivo activity by varying the steric hindrance at carbon atoms adjacent to the disulfide linkage.
abstract: In this report, we describe the synthesis of a panel of disulfide-linked huC242 (anti-CanAg) antibody maytansinoid conjugates (AMCs), which have varying levels of steric hindrance around the disulfide bond, in order to investigate the relationship between stability to reduction of the disulfide linker and antitumor activity of the conjugate in vivo. The conjugates were first tested for stability to reduction by dithiothreitol in vitro and for plasma stability in CD1 mice. It was found that the conjugates having the more sterically hindered disulfide linkages were more stable to reductive cleavage of the maytansinoid in both settings. When the panel of conjugates was tested for in vivo efficacy in two human colon cancer xenograft models in SCID mice, it was found that the conjugate with intermediate disulfide bond stability having two methyl groups on the maytansinoid side of the disulfide bond and no methyl groups on the linker side of the disulfide bond (huC242-SPDB-DM4) displayed the best efficacy. The ranking of in vivo efficacies of the conjugates was not predicted by their in vitro potencies, since all conjugates were highly active in vitro, including a huC242-SMCC-DM1 conjugate with a noncleavable linkage which showed only marginal activity in vivo. These data suggest that factors in addition to intrinsic conjugate potency and conjugate half-life in plasma influence the magnitude of antitumor activity observed for an AMC in vivo. We provide evidence that bystander killing of neighboring nontargeted tumor cells by diffusible cytotoxic metabolites produced from target cell processing of disulfide-linked antibody-maytansinoid conjugates may be one additional factor contributing to the activity of these conjugates in vivo.
authors.affiliation: ImmunoGen, Inc. , 830 Winter Street, Waltham, Massachusetts 02451, United States.
authors.forename: Brenda A
authors.initials: BA
authors.lastname: Kellogg
authors.forename: Lisa
authors.initials: L
authors.lastname: Garrett
authors.forename: Yelena
authors.initials: Y
authors.lastname: Kovtun
authors.forename: Katharine C
authors.initials: KC
authors.lastname: Lai
authors.forename: Barbara
authors.initials: B
authors.lastname: Leece
authors.forename: Michael
authors.initials: M
authors.lastname: Miller
authors.forename: Gillian
authors.initials: G
authors.lastname: Payne
authors.forename: Rita
authors.initials: R
authors.lastname: Steeves
authors.forename: Kathleen R
authors.initials: KR
authors.lastname: Whiteman
authors.forename: Wayne
authors.initials: W
authors.lastname: Widdison
authors.forename: Hongsheng
authors.initials: H
authors.lastname: Xie
authors.forename: Rajeeva
authors.initials: R
authors.lastname: Singh
authors.forename: Ravi V J
authors.initials: RV
authors.lastname: Chari
authors.forename: John M
authors.initials: JM
authors.lastname: Lambert
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Lutz
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood / Q000737:chemistry* / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000097:blood / Q000737:chemistry* / Q000494:pharmacology; D002244:Carbon / Q000737:chemistry*; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D004220:Disulfides / Q000097:blood / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D008453:Maytansine / Q000097:blood / Q000737:chemistry* / Q000494:pharmacology; D051379:Mice; D008815:Mice, Inbred Strains; D016513:Mice, SCID; D008968:Molecular Conformation; D023041:Xenograft Model Antitumor Assays
chemical_list: D000906:Antibodies; D000970:Antineoplastic Agents; D004220:Disulfides; D008453:Maytansine; D002244:Carbon