When tested in mice bearing established subcutaneous KB xenografts, dose-dependent antitumor activity was observed (Fig. IGNs represent a new chemical class of cytotoxic molecules with high in vitro potency (IC50 values in the low pmol/L range) toward cancer cells. If it was not, then the actus reus of arson was not present and no conviction for arson would be possible. Maloney, A. Wilhelm, R. Singh, Study supervision: M.L. Section 1734 solely to indicate this fact. Offences Against the Person Act 1861 s. 47, Criminal Appeal Act 1968 (c.19) s.33(2), Criminal Damage Act 1971 (c.48) s.1, Criminal Damage Act 1971 (c.48) s.1(1), Criminal Damage Act 1971 (c.48) s.1(2), Criminal Damage Act 1971 (c.48) s.4, Cruelty to Animals Act 1849 s.2, This page was last edited on 16 August 2020, at 10:40. This antibody–drug conjugate (ADC) technology has been validated following the FDA approval of: (i) brentuximab vedotin, a conjugate of an anti-CD30 antibody with the dolastatin derivative, monomethyl auristatin E (12) and (ii) ado-trastuzumab emtansine (T-DM1), a conjugate of trastuzumab with the potent maytansinoid DM1 (13, 14). These ADCs incorporate derivatives of the PBD dimer SJG-136, a highly toxic small molecule with a maximally tolerated dose (MTD) of ∼1.2 μg/kg in humans (19). To address these factors, we developed a new class of highly cytotoxic molecules, the IGNs, consisting of indolinobenzodiazepine pseudodimers for use as effector molecules for ADCs. In the treatment arm, deaths occurred in 2 of 4 animals. Lambert. 5A). A, scheme for the preparation of an IGN ADC (anti-FRα ADC 4a) with a disulfide linker. 5th Jul 2019 Bystander killing may play an important role clinically in the treatment of solid tumors that often express the antigen in a heterogeneous manner or where all tumor cells may not be accessible to the ADC. Nevertheless, the defendant was convicted for recklessly causing damage by omission. ADC 4a was considerably more active in vivo than 3c. 3B) was estimated to be about 1 mg/kg (20 μg/kg linked IGN) giving an in vivo therapeutic index (TI), defined as the ratio of MTD to MED, of about 10. In addition, incorporation of a phenyl group between the two monomer units provides a site for linker attachment. 1C), body weights in non–tumor-bearing CD-1 mice treated with monoimine IGN ADC, using the same antibody and schedule as 1c but at a two-fold higher dose (7.5 mg/kg, qw × 3), tracked with that of the vehicle control, with no evidence of prolonged or delayed toxicity (Fig. Mice received a single intravenous administration of vehicle (0.2 ml/mouse) or IGN ADC on day 1 (i.e., day 8 after tumor cell inoculation). For highly potent molecules, such as dolastatin-10, a tubulin-interacting agent, or adozelesin, a DNA alkylator, the MTD (3, 5) in humans is about 33% of the mouse LD10. Whiteman, E.E. Audette has ownership interest (including patents) in shares of stock. Reid, K.E. Cells were then pelleted, washed with PBS, and lysed in TRIS buffer (10 mmol/L, pH 8.5 containing 5 mmol/L EDTA, 0.2% SDS, 200 mmol/L NaCl, 0.1 mg/mL proteinase K) for 5 hours at 37°C. For example, it has been estimated that more than 99% of the cells within a tumor have to be killed to achieve a complete remission in the patient and significantly more to accomplish tumor eradication (1). In an in vivo model established using the human AML cell line HL-60 (Fig. Miller's defence was that there was no actus reus coinciding with mens rea. The anti-CD33-IGN ADC 1b killed the leukemia cell line NB4, with an IC50 value of 3 pmol/L, despite the low antigen expression on these cells (∼7,000 CD33 molecules/cell). This modification, coupled with linker optimization, generated ADCs that were well tolerated in mice and demonstrated robust antitumor activity in multiple tumor models at doses 1.5% to 3.5% of maximally tolerated levels. Do you have a 2:1 degree or higher? He fell asleep with a lit cigarette in his hand, which started a fire. https://en.wikipedia.org/w/index.php?title=R_v_Miller&oldid=973281187, Creative Commons Attribution-ShareAlike License. In contrast, ADC 3c with a noncleavable linker was incapable of killing bystander cells even when a high number of antigen-positive cells were added to the mixed culture. The anti-EGFR antibody used for ADC conjugation has no inherent antitumor activity. Lambert, R.V.J. While the two compounds co-purify with cellular DNA to a comparable final extent after 20 hours, the monoimine-containing IGN achieves only half the level of DNA modification as that achieved with the diimine at the earlier 5-hour time point (39 ± 2 vs. 75 ± 3 pmol adduct/μg DNA, respectively). Chari, Other (head of the R & D group guiding the research and Chief Science Officer during the time the research was conducted): J.M. Upon receipt, each line was expanded by passaging 2 to 3 times, aliquoted, and frozen. In this assay, a fixed number of antigen-negative cells are cocultured with varying number of antigen-positive cells and treated with the ADC. Mayo, A. Wilhelm, H.A. Overall, the findings of this study hold considerable promise for improving the success of the ADC technology and providing better therapeutic outcomes for personalized cancer treatments. ADCs with a cleavable (disulfide) linkage: The IGN dimer bearing a thiol or an imine-sulfonated thiol was treated in situ with SPDB or sulfo-SPDB in DMA, in the presence of a base (N-N-diisopropylethylamine) to provide the IGN with an activated ester. The Courtscheduled the four days between 5 and 8 December 2016 for the hearing. ADCs were prepared with three monoclonal antibodies that target cell surface antigens EpCAM, CD33, and FRα (Fig. injection, 0.5–2.5 mg/kg) in a subcutaneous xenograft model established with the human epidermoid carcinoma cell line KB in SCID mice. Modafferi, Analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis): M.L. Miller, N.E. C, in vivo antitumor activity of the anti-CD33 ADC 5b in comparison to a mixture of unconjugated IGN (4) plus the anti-CD33 antibody at equivalent doses to the ADC in a HL60 xenograft model. Her plea of diminished responsibility was accepted. The addition of an excess (1 μmol/L) of unconjugated antibody abrogated the cytotoxic effect demonstrating antigen-specific killing (Supplementary Fig. Mayo, J. Pinkas, R.V.J. All in vivo procedures were performed in strict accordance with the NIH Guide for the Care and Use of Laboratory Animals.