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Trastuzumab emtansine (T-DM1) is an antibody-drug conjugate consisting of the anti-HER2 antibody trastuzumab linked via a nonreducible thioether linker to the maytansinoid antitubulin agent DM1. T-DM1 has shown favorable safety and efficacy in patients with HER2-positive metastatic breast cancer. In previous animal studies, T-DM1 exhibited better pharmacokinetics (PK) and slightly more efficacy than several disulfide-linked versions. The efficacy findings are unique, as other disulfide-linked antibody-drug conjugates (ADC) have shown greater efficacy than thioether-linked designs. To explore this further, the in vitro and in vivo activity, PK, and target cell activation of T-DM1 and the disulfide-linked T-SPP-DM1 were examined. Both ADCs showed high in vitro potency, with T-DM1 displaying greater potency in two of four breast cancer cell lines. In vitro target cell processing of T-DM1 and T-SPP-DM1 produced lysine-N e -MCC-DM1, and lysine-N e -SPP-DM1 and DM1, respectively; in vivo studies confirmed these results. The in vitro processing rates for the two conjugate to their respective catabolites were similar. In vivo, the potencies of the conjugates were similar, and T-SPP-DM1 had a faster plasma clearance than T-DM1. Slower T-DM1 clearance translated to higher overall tumor concentrations (conjugate plus catabolites), but unexpectedly, similar levels of tumor catabolite. These results indicate that, although the ADC linker can have clear impact on the PK and the chemical nature of the catabolites formed, both linkers seem to offer the same payload delivery to the tumor.

Conjugation of cytotoxic compounds to antibodies that bind to cancer-specific antigens makes these drugs selective in killing cancer cells. However, many of the compounds used in such antibody-drug conjugates (ADC) are substrates for the multidrug transporter MDR1. To evade the MDR1-mediated resistance, we conjugated the highly cytotoxic maytansinoid DM1 to antibodies via the maleimidyl-based hydrophilic linker PEG 4 Mal. Following uptake into target cells, conjugates made with the PEG 4 Mal linker were processed to a cytotoxic metabolite that was retained by MDR1-expressing cells better than a metabolite of similar conjugates prepared with the nonpolar linker N-succinimidyl-4-(maleimidomethyl)cyclohexane-1-carboxylate (SMCC). In accord, PEG 4 Mal-linked conjugates were more potent in killing MDR1-expressing cells in culture. In addition, PEG 4 Mal-linked conjugates were markedly more effective in eradicating MDR1-expressing human xenograft tumors than SMCC-linked conjugates while being tolerated similarly, thus showing an improved therapeutic index. This study points the way to the development of ADCs that bypass multidrug resistance.

Grasslands have an underground biomass component that serves as a carbon (C) storage sink. Switchgrass (Panicum virgatum L.) has potential as a biofuel crop. Our objectives were to determine biomass and C partitioning in aboveground and belowground plant components and changes in soil organic C in switchgrass. Cultivars Sunburst and Dacotah were field grown over 3 yr at Mandan, ND. Aboveground biomass was sampled and separated into leaves, stems, senesced, and litter biomass. Root biomass to 1.1‐m depth and soil organic C to 0.9‐m depth was determined. Soil C loss from respiratory processes was determined by measuring CO2 flux from early May to late October. At seed ripe harvest, stem biomass accounted for 46% of total aboveground biomass, leaves 7%, senesced plant parts 43%, and litter 4%. Excluding crowns, root biomass averaged 27% of the total plant biomass and 84% when crown tissue was included with root biomass. Carbon partitioning among aboveground, crown, and root biomass showed that crown tissue contained approximately 50% of the total biomass C. Regression analysis indicated that soil organic C to 0.9‐m depth increased at the rate of 1.01 kg C m−2 yr−1 Carbon lost through soil respiration processes was equal to 44% of the C content of the total plant biomass. Although an amount equal to nearly half of the C captured in plant biomass during a year is lost through soil respiration, these results suggest that northern Great Plains switchgrass plantings have potential for storing a significant quantity of soil C.

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