Argos Therapeutics (United States)

<title>Abstract</title> This study investigates the effects of phytotoxins derived from <italic>Spathiphyllum walsii </italic> (peace lily) on the behavioral plasticity of <italic>Drosophila melanogaster</italic> larvae, focusing on their implications for sensory perception and motor function. Phytotoxins, known for their diverse bioactive properties, serve as both a defense mechanism for plants and potential therapeutic agents in medicine. The research involved the extraction of phytochemicals from <italic>Spathyiphyllum </italic>leaves, followed by a series of behavioral assays to assess larval responses to food stimuli, olfactory cues, and environmental conditions. The results demonstrated that larvae treated with <italic>SL </italic>extract exhibited significant delays in reaching food sources compared to control groups, indicating impaired sensory and motor functions. Notably, higher concentrations of the extract produced effects similar to those of colchicine, a well-known microtubule inhibitor. Behavioral assays revealed that both treatments disrupted normal larval behavior, suggesting that phytotoxins may influence neural circuits and microtubule dynamics. These results underscore the potential of plant-derived compounds as modulators of neural function and behavior, highlighting both their therapeutic prospects and risks associated with their use. This study highlights the potential of plant-derived compounds in understanding neurobiological processes and offers insights into their therapeutic applications. Further research is warranted to elucidate the mechanisms underlying these effects and to explore the broader implications of phytotoxins in pharmacotherapy and toxicology.

Neurofibromatosis type 1 (NF1) is a multi-system, autosomal dominant genetic disorder driven by the systemic loss of the NF1 protein neurofibromin. Loss of neurofibromin in Schwann cells is particularly detrimental, as the acquisition of a ‘second-hit’ (e.g., complete loss of NF1) can lead to the development of plexiform neurofibromas (pNF). pNFs are painful, disfiguring tumors with an approximately 1 in 5 chance of sarcoma transition. Selumetinib and mirdametinib are currently the only medicines approved by the U.S. Food and Drug Administration (FDA) for the treatment of pNFs. This motivates the need to develop new therapies, either derived to treat NF1 haploinsufficiency or complete loss of NF1 function. To identify new therapies, we need to understand the impact neurofibromin has on Schwann cells. Here, we aimed to characterize differences in high-content microscopy in neurofibromin-deficient Schwann cells. We applied a fluorescence microscopy assay (called Cell Painting) to an isogenic pair of Schwann cell lines (derived from ipn02.3 2λ), one of wildtype genotype (NF1+/+) and one of NF1 null genotype (NF1−/−). We modified the canonical Cell Painting assay to mark four organelles/subcellular compartments: nuclei, endoplasmic reticulum, mitochondria, and F-actin. We utilized CellProfiler to perform quality control, illumination correction, segmentation, and cell morphology feature extraction. We segmented 20,680 NF1 wildtype and null cells, measured 894 significant cell morphology features representing various organelle shapes and intensity patterns, and trained a logistic regression machine learning model to predict the NF1 genotype of single Schwann cells. The machine learning model had high performance, with training and testing data yielding a balanced accuracy of 0.85 and 0.80, respectively. However, when applied to a new pair of Schwann cells, the model’s balanced accuracy dropped to 0.5, which is no better than random chance. This performance decline appears to result from morphology differences introduced by non-biological factors (cloning procedures, origin of parental cell line, and CRISPR procedures) of the second cell line pair. We plan to improve upon this preliminary model by refining the NF1 morphology signature using a broader panel of Schwann cell lines. Our goal is to apply this enhanced signature in large-scale drug screens of NF1-deficient cells to identify candidate therapeutic agents that specifically reverse the disease-associated morphology. Ultimately, we aim to identify agents that restore NF1 patient-derived Schwann cells to a phenotype resembling the NF1 wild-type and healthier state.

BackgroundPD-1/PD-L1 antibody-based therapies have demonstrated tremendous success in the treatment of a variety of cancers. However, these antibody drugs are associated with several disadvantages, such as weak tumor penetration, immune-related adverse events (irAEs) due to their long half-life and development of anti-drug antibodies. Here, we report the discovery of ALG-093989, a highly potent and orally bioavailable PD-L1 small molecule inhibitor, that may overcome the limitation of PD-1/PD-L1 antibodies.MethodsThe biochemical interaction of PD-1/PD-L1 and PD-L1 dimerization was assessed by AlphaLISA®. Cellular activity was measured using a co-culture assay of PD-1 expressing Jurkat NFAT luciferase T cells with PD-L1 expressing CHO cells. Pharmacokinetic (PK) studies were performed in rat. In vivo PD-L1 target occupancy was assessed at 6 hours following a single oral dose in a humanized-PD-L1 MC38 subcutaneous mouse model.ResultsALG-093989 inhibits PD-1/PD-L1 interaction with an IC50 of 14 pM and induces PD-L1 dimerization with an EC50 of 13 nM. ALG-093989 has similar T-cell activation potency as durvalumab, and approximately 10-fold improved T-cell activation potency vs. INCB086550, a PD-L1 small molecule inhibitor that demonstrated clinical response in a phase I study. Oral bioavailability of ALG-093989 in rat following a single dose at 15 mg/kg dosing was 40%. In the in vivo humanized-PD-L1 MC38 mouse model, a single oral dose of ALG-093989 at 5 mg/kg demonstrated similar PD-L1 target occupancy to that of 150 mg/kg orally-administered INCB086550 and 5 mg/kg intravenously-administered durvalumab.ConclusionsWe have discovered ALG-093989, a highly potent PD-L1 small molecule inhibitor, which shows similar T-cell activation potency as durvalumab, and ~ 10-fold improved T-cell activation potency vs. INCB086550. ALG-093989 has the same target occupancy in mice following oral dosing at a 30-fold lower dose than INCB03989. The properties of ALG-093989 warrant further evaluation as a potential candidate for drug development.

Chronic hepatitis B is the most important cause of liver cancer worldwide and affects more than 290 million people. Current treatments are mostly suppressive and rarely lead to a cure. Therefore, there is a need for novel and curative drugs that target the host or the causative agent, hepatitis B virus itself. Capsid assembly modulators are an interesting class of antiviral molecules that may one day become part of curative treatment regimens for chronic hepatitis B. Here we explore the characteristics of a particularly interesting subclass of capsid assembly modulators. These so-called non-HAP CAM-As have intriguing properties in cell culture but also clear virus-infected cells from the mouse liver in a gradual and sustained way. We believe they represent a considerable improvement over previously reported molecules and may one day be part of curative treatment combinations for chronic hepatitis B.

pydicom v1.3.0

31 Background: AGS-003 is an immunotherapy consisting of autologous dendritic cells electroporated with amplified total tumor RNA plus synthetic CD40L RNA and is currently being tested in combination with standard of care to extend survival of newly diagnosed metastatic renal cell carcinoma (RCC) patients in the Phase 3 ADAPT clinical trial. We set out to establish a model system based on the Renca mouse model to more thoroughly study the AGS-003 mechanism of action and identify optimal combination therapies, including with a PD1 checkpoint inhibitor (aPD1 CPI). Methods: Mouse DC precursors were processed in a similar manner to how human monocytes are processed to manufacture AGS-003. Mature DCs were injected s.c. to treat BALB/c mice in an orthotopic syngeneic RCC model. This model system was utilized to test the efficacy and mechanism of action of the AGS-003-like homologous mouse DCs in combination with a murine monoclonal antibody PD1 CPI (anti-mPD1). Results: Murine DCs with similar properties to AGS-003 in combination with anti-mPD1 and sunitinib, resulted in recruitment and migration of lymphocytes into the tumor microenvironment and an increase in peripheral blood CD8+CD28+CD45RA- memory T cells in vivo. Multiple combination dosing strategies were tested and only one dosing regimen (DCs administered prophylactically followed by aPD1 mAb administered therapeutically) showed a substantial synergistic effect (67 days median OS) while all other DC/aPD1 dosing combinations did not exceed the efficacy of DC monotherapy treatment (44.5-48 days median OS). Control animals treated with PBS had a median OS of 29 days. Conclusions: These data demonstrate the importance of the administration sequence for active immunotherapy and aPD1 CPI combination therapy. Our data suggest that the cellular immune response must be initiated and established prior to administration of the aPD1 CPI/sunitinib combination therapy in order to observe synergy in this RCC mouse model. This model may be useful to explore additional combination therapies and effective treatment regimens with other therapeutic agents to guide future clinical development.

357 Background: During the past decade, VEGF targeted therapies have become standard treatment for advanced RCC. While targeted therapies have yielded improved efficacy, durable remissions are rare, particularly in unfavorable risk subjects. In a pivotal trial, treatment with sunitinib yielded a median OS of 5.3 months for poor risk and 20.7 months for intermediate risk subjects. Similarly, the validation dataset for the Heng risk model (ASCO 2011) revealed a median OS of 8 months for poor risk and 21 months for intermediate risk patients treated with VEGF targeted therapies. Durable responses have been elicited by immunotherapy in RCC. We report an update on patients with mRCC treated with combined VEGF TKI (sunitinib) plus autologous immunotherapy (AGS-003). Methods: In this phase II study, subjects with unfavorable prognosis mRCC (poor and intermediate risk) were treated with sunitinib plus autologous dendritic cell immunotherapy (AGS-003). Treatment consisted of standard 6-week cycles of sunitinib plus AGS-003 (once every 3 weeks x5 doses, then every 12 weeks until PD). Tumor response was assessed per RECIST and subjects were followed for PFS and OS. Immune monitoring samples were taken at screening, baseline and after the third and fifth doses of AGS-003 and analyzed by multiparametric flow cytometry. Results: 21 subjects were treated. As previously reported, the median overall PFS was 11.2 months and the final median OS was 30.2 months. When analyzed by baseline Heng risk status, the median PFS was 19.4 months for intermediate (n=11) and 5.8 months for subjects with poor risk features (n=10) at baseline. The median OS is 39.5+ months for intermediate and 9.1 months for subjects with poor risk. Conclusions: The results from this single-arm phase II study represent a near doubling of expected PFS and OS for unfavorable risk subjects treated with AGS-003 plus sunitinib. Updated survival and immunologic findings will be presented, as 8 of 21 subjects are still alive and continue to be followed. These results support the ongoing phase III ADAPT study, which is designed to validate these encouraging clinical and immunologic findings. Clinical trial information: NCT00678119.