Supplementary Fig. S1 from Engineered anti-CD70 antibody-drug conjugate with increased therapeutic index

Supplementary Table S4 from Engineered anti-CD70 antibody-drug conjugate with increased therapeutic index

Plasma stability of quaternary ammonium linker in rodent plasma.

Antibody humanization, binding assays, conjugation, mass spec, additional in vivo details

Pharmacokinetic profile of clearance for three ADCs.

Legends of supplementary Figure S1, S2 and S3

Supplementary Fig. S2 from Anti-CD30 diabody-drug conjugates with potent antitumor activity

Experimental details for the chemical synthesis of the drug-linkers.

SGN-35C is an antibody drug conjugate composed of an anti-CD30 antibody conjugated to a camptothecin-derived topoisomerase 1 (TOP1) inhibitor payload. SGN-35C was designed to leverage the antibody backbone from brentuximab vedotin (BV) and the novel mechanism of action of camptothecin-derived ADCs. Here, we evaluated the mechanism of action, cytotoxicity, and safety in in vitro and in vivo models. To characterize the mechanism of action of SGN-35C in vitro and in vivo experiments were performed in CD30-positive anaplastic large cell lymphoma (ALCL), Hodgkin lymphoma (HL) cell lines, and versions of these cell lines that are resistant to BV. SGN-35C binding, internalization, induction of DNA damage, cell cycle analysis, and in vitro cytotoxicity were evaluated. To assess if payload released in CD30-positive cells could kill adjacent CD30-negative cells, an in vitro bystander assay of CD30-positive and CD30-negative co-cultures was developed. The in vivo antitumor activity of SGN-35C was evaluated in animals bearing subcutaneous lymphoma models. SGN-35C antitumor activity was evaluated in a xenograft model of heterogeneous CD30 expression consisting of a mixture of a BV-resistant Karpas-299 cell line mixed with parental Karpas-299 cells to assess bystander activity in vivo. The tolerability of SGN-35C was assessed in a non-human primate model. SGN-35C binding to CD30-positive cells was similar to the unconjugated anti-CD30 antibody and demonstrated internalization into CD30-expressing cells. Treatment of cells with SGN-35C for 24 hours led to an increase of the DNA damage marker phospho-γH2A.X, which induced an increase in the fraction of cells in S-phase, consistent with the mechanism of camptothecin-based TOP1 inhibitors. Cytotoxicity was observed in ALCL and HL cell lines treated with SGN-35C, including an ALCL cell line resistant to BV. In a mixture of CD30-positive and CD30-negative cells, SGN-35C treatment induced cytotoxicity of the CD30-negative cells, demonstrating that SGN-35C can elicit bystander activity. A single dose of SGN-35C led to significant tumor growth inhibition in animals bearing lymphoma xenografts, including a BV-resistant model and a model of heterogeneous CD30 expression, demonstrating that SGN-35C can elicit bystander activity in vivo. SGN-35C demonstrates a tolerability profile in non-human primates that is consistent with camptothecins. A Phase 1, first-in-human study is planned to evaluate the safety and anti-tumor activity of SGN-35C in lymphoid malignancies.

Supplementary Table S3 from Engineered anti-CD70 antibody-drug conjugate with increased therapeutic index

S1. Antitumor activity and intratumoral drug release by hIgG-vcMMAE. S2: Depleting TAMs attenuate the activity of hIgG-vcMMAE. S3. Impaired FcgR binding with G1V1 variant

In vitro dose response curves

Supplementary Fig. S2 from Engineered anti-CD70 antibody-drug conjugate with increased therapeutic index

Supplementary Material from Engineered anti-CD70 antibody-drug conjugate with increased therapeutic index

Gating strategy, primary AML cytotoxicity assay,additional cell liens for pH2AX assay and caspase assay, and h7G3ec antibody sequence

Supplementary Fig. S1 from Anti-CD30 diabody-drug conjugates with potent antitumor activity

Supplementary Material from Anti-CD30 diabody-drug conjugates with potent antitumor activity

The Front Cover shows the antimitotic tubulysin M bound to bovine brain tubulin. Highlighted in yellow is the labile C11 acetate, the loss of which leads to greatly reduced biochemical activity. The corresponding article interrogates the effect of drug-linker chemistry and conjugation site on the stability of this acetate and the in vivo activity of antibody–drug conjugates (ADCs) delivering tubulysin M. The results led the team from a protease cleavable 4-load ADC (top) to an optimized β-glucuronidase cleavable 2-load ADC (bottom). More information can be found in the Communication by Joseph Z. Hamilton et al.