CD205 is a type I transmembrane glycoprotein and is a member of the C-type lectin receptor family. Analysis by mass spectrometry revealed that CD205 was robustly expressed and highly prevalent in a variety of solid malignancies from different histotypes. IHC confirmed the increased expression of CD205 in pancreatic, bladder, and triple-negative breast cancer (TNBC) compared with that in the corresponding normal tissues. Using immunofluorescence microscopy, rapid internalization of the CD205 antigen was observed. These results supported the development of MEN1309/OBT076, a fully humanized CD205-targeting mAb conjugated to DM4, a potent maytansinoid derivate, via a cleavable N-succinimidyl-4-(2-pyridyldithio) butanoate linker. MEN1309/OBT076 was characterized in vitro for target binding affinity, mechanism of action, and cytotoxic activity against a panel of cancer cell lines. MEN1309/OBT076 displayed selective and potent cyto-toxic effects against tumor cells exhibiting strong and low to moderate CD205 expression. In vivo, MEN1309/OBT076 showed potent antitumor activity resulting in durable responses and complete tumor regressions in many TNBC, pancreatic, and bladder cancer cell line-derived and patientderived xenograft models, independent of antigen expression levels. Finally, the pharmacokinetics and pharmacodynamic profile of MEN1309/OBT076 was characterized in pancreatic tumor-bearing mice, demonstrating that the serum level of antibody-drug conjugate (ADC) achieved through dosing was consistent with the kinetics of its antitumor activity. Overall, our data demonstrate that MEN1309/OBT076 is a novel and selective ADC with potent activity against CD205-positive tumors. These data supported the clinical development of MEN1309/OBT076, and further evaluation of this ADC is currently ongoing in the first-inhuman SHUTTLE clinical trial.

Membrane technology without a phase-change process has demonstrated its low-cost, high-energy-efficiency advantages. High fractional free volume polymers of intrinsic microporosity (PIMs), such as poly(1-trimethylsilyl-1-propyne) (PTMSP), are known for high permeability but disappointing gas selectivity. Porous aromatic frameworks and hyper-cross-linked polymers (HCPs) are effective membrane additives; however, prior works have examined their performance through chemical interactions and other material characteristics. Using a recently developed facile and scalable controlled HCP synthesis method, the size effect of these additives can now be investigated. In this work, HCPs with gradually increasing molecular weight/particle size were combined to investigate the performance and mixing/dispersion behavior in PTMSP mixed-matrix membranes. Unexpectedly, the largest dispersible additive (V125E) resulted in a membrane with drastically higher selectivity (H 2 /CH 4 = +690%) and similar H 2 permeability, which is a performance above the 2015 upper bound for H 2 gas pairs. With identical additive affinity/compatibility, we demonstrate the benefit of size-controlled HCP additives for enhancing membrane performance.

Antibody-drug conjugates (ADCs) are a recent and exciting development for targeted therapy of cancer. Their efficacy is governed by ADC-intrinsic characteristics such as avidity, drug load and linker chemistry, and mechanisms of activation and action, which can be controlled or clarified in the early stages of ADC development. In contrast, the properties that define a promising ADC target are still somewhat unclear. OGAP is a unique proteomic database that integrates information at the tissue, disease and protein isoform level across diseases, indications, and normal tissues to clarify protein expression levels and profiles. Specifically, it currently holds information on ∼2,000,000 human protein peptide sequences, ∼16,000 human proteins sequenced, ∼7,000 cancer membrane proteins, ∼50 tissues/organs, and ∼60 diseases. Building on OGAP and a proprietary sample preparation and processing workflow that relies on state-of-the-art high-throughput mass spectrometry and data processing to provide quantitative information on over 4,000 membrane-enriched proteins from ∼ 15,000 unique peptide sequences per analysis, we have established a novel predictive tool to establish each protein's potential to serve as a target for ADC development. The tool considers proteomic and target-specific information on antigenicity, structure, function, expression level, regulation, and tissue distribution in order to highlight the most suitable candidates for ADC development. We will demonstrate the utility of this process for the protein family of G-protein coupled receptors (GPCRs), which according to a recent bioinformatics prediction encompasses 899 distinct members in the human genome. These cell surface receptors are the target of more than one third of conventional drugs, yet their potential for ADCs is largely unexplored. Here we show that proteomics in the context of the OGAP database can highlight which of this large family of receptors have the potential to become true ADC targets. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3869. doi:1538-7445.AM2012-3869

<p>Supplementary Table S5: Microscopic IHC staining for CD205 in different cynomolgus tissues after treatment with MEN1309/OBT076. Staining: 1, minimal (+, rare cells with low staining intensity); 2, slight/moderate (++, many cells with medium/high staining intensity); 3, marked (+++, significant numbers of cells with high staining intensity); 0, negative (no stained cells); -, no background present; tm, tissue missing; lp, lesion present; p, background staining present; N/A, not applicable.</p>

<p>Supplementary Table S4: Antitumor activity of MEN1309/OBT076 on xenograft and PDX models with different CD205 antigen expression. a) PDX models.b) Score indicate the staining intensity. HE = heterogeneous. HO = homogeneous. c) TVI%, tumor volume inhibition, expressed as a percentage, indicates the ratio % between the mean of the tumor volume of the test article-treated group and the control treated with drug vehicle calculated at the Nadir of tumor growth.</p>

<p>Supplementary Table S3: Cross reactivity data obtained from whole blood of different species. Values are expressed as the concentration at which MBH1309/OBT076 has half-maximum binding [EC50].</p>

<p>Supplementary Figure S1. Representative images of IHC staining for CD205 in different human and cynomolgus tissues, obtained with 2 μg/ml of MEN1309/OBT076 at 20X and 10X magnification. 1) Human donor spleen; moderate to strong membranous/granular cytoplasmic staining of several mononuclear cells present in the white pulp of the spleen (circle); 2) Cynomolgus donor spleen; the staining intensity and distribution pattern was very similar to that described in human samples; 3) Human donor lymph node; a moderate to marked membranous/cytoplasmic staining of mononuclear cells in the cortex, para-cortex and germinal centers of lymphoid follicles was observed; 4) Cynomolgus donor lymph node; positive mononuclear cells presented a moderate to strong membranous/granular cytoplasmic staining; 5) Human donor breast; a concentration-dependent cytoplasmic staining of epithelial cells in glandular alveoli (rectangle) and ducts (circle); 6) Cynomolgus donor breast; the staining was weaker in intensity when compared to human donors, but the pattern was comparable; 7) Human donor colon; membranous/cytoplasmic signal of a variable number of mononuclear cells was observed in the lamina propria and GALT (circle). Minimal to slight cytoplasmic staining of the superficial mucosal epithelium (rectangle); 8) Cynomolgus donor colon; a variable number of mononuclear cells located in lamina propria and GALT (circle) are positive with membranous/cytoplasmic signal; 9) Human donor ureter; a minimal but concentration-dependent staining was observed in mucosal epithelium (rectangle). Rare mononuclear cells with positive signal in lamina propria (circles); 10) Cynomolgus donor ureter; occasional mononuclear cells with membranous/granular cytoplasmic staining was observed in the lamina propria; 11) Human donor cervix; a concentration-dependent cytoplasmic staining of cervical gland epithelium was observed (circle). Cytoplasmic staining of the basal cell layer of mucosal epithelium was also observed (rectangle); 12) Cynomolgus donor cervix; positive mononuclear cells with a membranous/granular cytoplasmic staining were present in the mucosa-associated lymphoid nodules (circle).</p>

<p>Supplementary Table S1. Commercial tumor TMAs representing different solid tumor types. Supplementary Table S2. Summary of CD205 expression in human solid tumor types.</p>