Preliminary Safety Evaluation Research Articles

Protocol of investigations for the preliminary assessment of the structural safety of highway overpasses

This paper presents the results of a protocol of investigations that have been carried out on 15 overpasses, located along the highway A18 in Sicily (south of Italy) and built about fifty years ago, with the aim of achieving a preliminary evaluation of the structural safety of these overpasses. The investigated overpasses are characterized by a three-span deck consisting of a reinforced concrete slab and prestressed concrete girders. The girders of the central span are simply supported by two Gerber saddles located on cantilever girders at the two ends of the same span. The safety level of the overpasses is investigated by means of different activities that include visual inspection and computation of the class of attention of the overpasses according to the Italian Bridge Guidelines for the classification and the risk management, safety evaluation and monitoring of existing bridges, laboratory and in situ tests on materials, pachometric, georadar and video-endoscopic tests on structural details, and static load tests of the overpasses. The results of the static load tests have also been used to assess the accuracy of two numerical models of the overpasses. These models are elastic but characterised by different levels of accuracy and computational burden: while the first model is the well-established and simple Courbon-Albenga method, the second is a finite element model characterised by a more refined simulation of the response of slabs, longitudinal girders and crossbeams. The results of the investigations have been critically analysed and discussed to underline the ability of the protocol to reveal possible deficiencies of the overpasses and thus identify overpasses that are liable to structural interventions or more accurate and expensive investigations.

The first-in-class bispecific antibody IBI318 (LY3434172) targeting PD-1 and PD-L1 in patients with advanced tumors: a phase Ia/Ib study

BackgroundThere is an unmet clinical need to enhance the response rate and safety of anti-PD-1/PD-L1-based cancer immunotherapy (IO). Herein, we presented the clinical study of IBI318 (LY3434172), a first-in-class bispecific antibody (bsAb) targeting PD-1 and PD-L1, in patients with advanced tumors.MethodsIn this open-label, multicenter Phase Ia/Ib study of IBI318, the Phase Ia involved dose escalation and a safety dose expansion, while the Phase Ib focused on preliminary safety and efficacy evaluation in non-small cell lung cancer (NSCLC) and nasopharyngeal carcinoma (NPC). In Phase Ia, patients with advanced tumors received IBI318 doses ranging from 0.3 to 1200 mg every two weeks (Q2W) to determine the recommended Phase 2 dose (RP2D). In Phase Ib, NSCLC or NPC patients from five cohorts with varying treatment histories received IBI318 at the RP2D. The primary endpoint was safety and the secondary endpoints included efficacy assessed by investigators according to RECIST v1.1, pharmacokinetics, immunogenicity, and pharmacodynamics.ResultsFrom February 11, 2019, to January 25, 2022, a total of 103 eligible patients were enrolled (Phase Ia, n = 55; Phase Ib, n = 48). The median follow-up was 10.1 months (range 0.7–28.6). The RP2D was determined to be 300 mg Q2W. Treatment-related adverse events (TRAEs) of any grades occurred in 88 patients (85.4%), while 10 patients (9.7%) experienced grade ≥ 3 TRAEs. The objective response rate (ORR) was 15.5% and the disease control rate (DCR) was 49.5% in all patients. In Phase Ib, the confirmed ORR was 45.5% in treatment-naïve NSCLC patients and 30.0% in IO-naïve NPC patients who had failed or were intolerant to platinum-based treatments.ConclusionsIBI318 demonstrated a favorable safety profile and preliminary efficacy in treatment-naïve NSCLC and IO-naïve NPC patients. Further clinical studies are needed to assess the full therapeutic potential of PD-1/PD-L1 dual inhibition with bsAbs.

Liquid chromatography‐tandem mass spectrometry (LC‐MS/MS)‐based quantification of hydroxyanthracene derivatives in Aloe vera (L.) Burm. f. gel commercial beverages and preliminary safety evaluation through in vitro genotoxicity studies

Liquid chromatography‐tandem mass spectrometry (LC‐MS/MS)‐based quantification of hydroxyanthracene derivatives in <i>Aloe vera</i> (L.) Burm. f. gel commercial beverages and preliminary safety evaluation through in vitro genotoxicity studies

Synergistic Enhancement of Diagnostic Imaging: Synthesis and Preliminary Safety Evaluation of Gadolinium-Doped Carbon Quantum Dots as Dual-Contrast Agent.

The present study explores the synthesis and bio-safety evaluation of gadolinium-doped carbon quantum dots (GCQDs) as a potential dual-contrast agent for diagnostic imaging. GCQDs exhibit both fluorescent and magnetic properties, making them suitable for UV-Vis and magnetic resonance imaging (MRI). The synthesis of GCQDs was achieved via hydrothermal treatment, incorporating gadolinium into the carbon quantum dot matrix. The magnetic properties of GCQDs were analyzed, showing significantly enhanced values compared to gadobutrol, a common MRI contrast agent. However, synthesis constraints limit the gadolinium content achievable in nanodots. To assess the safety of GCQDs, their effects on the embryonic development of zebrafish (Danio rerio) were examined. Various concentrations of GCQDs were tested, observing mortality rates, hatchability, malformations, heartbeats, spontaneous movement, and GCQDs uptake. Dialysis studies indicated that gadolinium ions are incorporated into the internal structure of the carbon nanodots. Zebrafish toxicity tests revealed that while survival rates were comparable to control groups, hatchability decreased significantly with higher gadolinium concentrations in GCQDs. Fluorescence microscopy showed no statistical differences in the fluorescence intensity between groups. These findings suggest that GCQDs could serve as an effective dual-contrast agent, combining the optical imaging capabilities of CQDs with the enhanced MRI contrast provided by gadolinium. This study underscores the need for further research on the synthesis methods and biological interactions of GCQDs to ensure their safety and efficacy in medical applications.

Reactive oxygen species responsive double-locked liposome collaborative photodynamic therapy for reducing electrical conduction recurrence after radiofrequency catheter ablation

Radiofrequency catheter ablation (RFCA) is the preferred technique for the treatment of atrial fibrillation, but the recovery of electrical conduction after ablation seriously endangers the health of patients. This study aimed to develop reactive oxygen species (ROS) responsive double-locked liposome collaborative photodynamic therapy (PDT) to target the ablation area and reduce the recovery of electrical conduction after ablation. The successful synthesis of β-cyclodextrin modified with phenylboronic acid pinacol ester (OCD) was confirmed by 1H NMR and FT-IR. Furthermore, the successful synthesis of octadecylamine-modified indocyanine green (ICG-ODA) was confirmed by 1H NMR and mass spectrometry. The ICG-ODA was encapsulated in liposomes to generate a double-locked hybrid liposome (ICG-ODA@rNP), which was subsequently characterized. Several properties of ICG-ODA@rNP were evaluated, including the drug release, targeting ability and ability to inhibit electrical conduction recurrence. Moreover, a model was constructed for the blockage of electrical conduction after RFCA in rabbits to further evaluate ICG-ODA@rNP. The preliminary safety evaluation of ICG-ODA@rNP was also performed. The ICG-ODA@rNP with a uniform particle size showed excellent storage stability. The nanoparticle can sensitively release drugs under ROS environment, and exhibits excellent photothermal effects. Furthermore, ICG-ODA@rNP can circulate for a long time in vivo and accumulate significantly in the ablation area. In a pacing test with a left atrial appendage (LAA), these nanoparticles, combined with PDT, reduced the ratio of electrical conduction recovery, which was confirmed by a hematoxylin and eosin (H&E) test. Further molecular analysis revealed that ICG-ODA@rNP could increase RFCA-induced apoptosis and ROS levels. Specifically, ICG-ODA@rNP significantly increased the expression of Bax and cleaved caspase-3, and decreased the expression of Bcl-2. In addition, the excellent biosafety of the double-locked nanoparticle was verified. This study provides evidence that ICG-ODA@rNP, with the double lock characteristic and biosafety, which exhibits a targeting effect on RFCA-induced cardiac injury areas, which further reduce electrical conduction recovery in RFCA areas by collaborativing PDT.

The STAT3 inhibitor B9 alleviates lipopolysaccharide-induced acute lung injury through its anti-inflammatory effects

The development of acute lung injury (ALI), a common respiratory condition with multiple causes, is significantly influenced by the pro-inflammatory environment of signal transducer and activator of transcription 3 (STAT3) in macrophages. Our study aimed to evaluate the anti-inflammatory effects of B9 (N-(4-hydroxyphenyl)-9, 10-dioxo-9, 10-dihydroanthracene-2-sulfonamide), a novel inhibitor targeting the STAT3 SH2 domain, in macrophages and to assess its therapeutic potential for ALI using a mouse model of lipopolysaccharide (LPS)-induced ALI. We found that B9 (30 mg/kg) significantly reduced lung pathological damage and neutrophil infiltration caused by the intratracheal administration of LPS. Additionally, the high expression of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) in alveolar lavage fluid was also inhibited by B9 treatment. The decreased expression of CD86 and increased CD206 in lung tissue demonstrated the anti-inflammatory effect of B9, which was due to its inhibition of the STAT3 signaling pathway in macrophages of ALI mice. Furthermore, B9 suppressed the activation of RAW264.7 cells induced by LPS, characterized by its ability to inhibit the activation of iNOS and STAT3 in a dose-dependent manner, as well as reduce the secretion of IL-6 and IL-1β. The in vivo preliminary safety evaluation indicated that B9 had a favorable safety profile at the administered doses. These results suggest that B9 exerts a therapeutic effect on LPS-induced ALI, potentially by preventing the phosphorylation of STAT3 Y705 and S727 without affecting the STAT3 protein level. Taken together, these findings provide a foundation for developing B9 as a novel anti-inflammatory agent for ameliorating LPS-induced ALI.

Abstract 603: BB5523, a multiple kinase inhibitor with HPK1, VEGFR2 and TNIK inhibition, enhanced antitumor immunity in tumor models

Abstract Introduction: Immuno-suppressive micro-environment and abnormal signaling pathways are crucial for the proliferation of tumor cells. Hematopoietic progenitor kinase 1 (HPK1) inhibitor plays important roles in activation of exhausted T cells and enhancement of anti-tumor immune responses. VEGF/VEGFR2 pathway is the central therapeutic target in anti-angiogenic treatment in multiple cancers. Traf2- and Nck-interacting kinase (TNIK) is a member of the germinal center kinase family, and is reported to be essential for Wnt signaling and colorectal cancer (CRC) proliferation and progression. Blockade of critical kinases in multiple signaling pathways inhibits the proliferation, invasion and metastasis of cancer cells. Therefore, intergradation of these functions in a small molecule serves as the next generation of immune-oncology small molecules for cancer therapy. Methods: BB5523 was developed through structure-based drug design, and optimized by SAR analysis and medicinal chemistry iteration. Biochemical assays and cell-base functional assays were applied in compound evaluation. Pharmacokinetic (PK) and preliminary toxicity studies were performed with mice. CDX and syngeneic tumor models in mice were conducted to evaluate the in vivo efficacy of BB5523 as a single agent. Results: BB5523 shows high inhibition potency on HPK1 with good selectivity against other TCR signaling-related kinases, such as FYN and ZAP70. Inhibition of HPK1 kinase activity (IC50 = 3.3 nM) by BB5523 strongly suppresses the phosphorylation of the downstream biomarker protein SLP-76 (IC50 = 30 nM); and boosts the IL-2 secretion in purified human T cells and human peripheral blood mononuclear cells. BB5523 also shows good activities on VEGFR2 (IC50 = 1.6 nM) and TNIK (IC50 = 0.7 nM) which are crucial for the proliferation of cancer cells. In in vitro experiments, BB5523 significantly inhibits the proliferation of various tumor cell lines at sub-micromolar concentrations. PK profile of BB5523 shows good oral bioavailability (F &amp;gt; 80%). In preliminary safety evaluation, no abnormal symptoms were observed in mice dosed up to 100 mpk for 14 days. BB5523 showed significant tumor-growth inhibition efficacy in TE-8, JIMT-1, B16-F10 and MC38 tumor models. Conclusion: As a potent and oral available small molecule kinase inhibitor, BB5523 boosts anti-tumor immunity through HPK1 inhibition, while also blocking crucial signaling pathways in tumor micro-environment. BB5523 shows promising in vitro and in vivo efficacy with a well-tolerated safety profile. The preclinical study of BB5523 is in progress. Citation Format: Gongping Duan, Min Li, Puyong Zhang, Wei Hua, Enlun Hao, Shipeng Wang, Xiaoyi Ma, Junheng Wang, Lijie Wei, Xingmin Zhang. BB5523, a multiple kinase inhibitor with HPK1, VEGFR2 and TNIK inhibition, enhanced antitumor immunity in tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 603.

Characterization and Preliminary Safety Evaluation of Akkermansia muciniphila PROBIO

In addition to providing certain health advantages to the host, a bacterial strain must possess a clearly defined safety profile to be regarded as a probiotic. In this study, we present a thorough and methodical assessment of the safety of a novel strain of bacteria, Akkermansia muciniphila PROBIO, which was isolated from human feces. Firstly, we examined the strain’s overall features, such as its gastrointestinal tolerance and its physiological and biochemical traits. Next, we verified its genotoxic properties through bacterial reverse mutation and in vitro mammalian cell micronucleus assays. The drug sensitivity of A. muciniphila PROBIO was subsequently examined through an analysis of its antibiotic resistance genes. Additionally, the toxicological impact was verified through acute and sub-chronic toxicity studies. A genome-based safety assessment was conducted to gain further insights into gene function, including potential virulence factors and pathogenic properties. Finally, we assessed whether moxifloxacin resistance in A. muciniphila PROBIO is transferred using in vitro conjugation experiments. A. muciniphila PROBIO exhibited superior gastrointestinal tolerance, with no observed hematological or histopathological abnormalities. Moreover, the outcomes pertaining to mutagenic, clastogenic, or toxic impacts were found to be negative, even at exceedingly high dosages. Moreover, no adverse effects associated with the test substance were observed during the examination of acute and sub-chronic toxicity. Consequently, it was plausible to estimate the no-observed-adverse-effect level (NOAEL) to be 6.4 × 1011 viable bacteria for an average individual weighing 70 kg. Additionally, only three potential drug resistance genes and one virulence factor gene were annotated. A. muciniphila PROBIO is naturally resistant to moxifloxacin, and resistance does not transfer. Collectively, the data presented herein substantiate the presumed safety of A. muciniphila PROBIO for its application in food.

Preliminary evaluation of safety and migration of immune activated mesenchymal stromal cells administered by subconjunctival injection for equine recurrent uveitis.

Equine recurrent uveitis (ERU), an immune mediated disease characterized by repeated episodes of intra-ocular inflammation, affects 25% of horses in the USA and is the most common cause of glaucoma, cataracts, and blindness. Mesenchymal stromal cells (MSCs) have immunomodulatory properties, which are upregulated by preconditioning with toll-like receptor agonists. The objective was to evaluate safety and migration of TLR-3 agonist polyinosinic, polycytidylic acid (pIC)-activated MSCs injected subconjunctivally in healthy horses prior to clinical application in horses with ERU. We hypothesized that activated allogeneic MSCs injected subconjunctivally would not induce ocular or systemic inflammation and would remain in the conjunctiva for >14 days. Bulbar subconjunctiva of two horses was injected with 10 × 106 pIC-activated (10 μg/mL, 2 h) GFP-labeled MSCs from one donor three times at two-week intervals. Vehicle (saline) control was injected in the contralateral conjunctiva. Horses received physical and ophthalmic exams [slit lamp biomicroscopy, rebound tonometry, fundic examination, and semiquantitative preclinical ocular toxicology scoring (SPOTS)] every 1-3 days. Systemic inflammation was assessed via CBC, fibrinogen, and serum amyloid A (SAA). Horses were euthanized 14 days following final injection. Full necropsy and histopathology were performed to examine ocular tissues and 36 systemic organs for MSC presence via IVIS Spectrum. Anti-GFP immunohistochemistry was performed on ocular tissues. No change in physical examinations was noted. Bloodwork revealed fibrinogen 100-300 mg/dL (ref 100-400) and SAA 0-25 μg/mL (ref 0-20). Ocular effects of the subjconjucntival injection were similar between MSC and control eyes on SPOTS grading system, with conjunctival hypermia, chemosis and ocular discharge noted bilaterally, which improved without intervention within 14 days. All other ocular parameters were unaffected throughout the study. Necropsy and histopathology revealed no evidence of systemic inflammation. Ocular histopathology was similar between MSC and control eyes. Fluorescent imaging analysis did not locate MSCs. Immunohistochemistry did not identify intact MSCs in the conjunctiva, but GFP-labeled cellular components were present in conjunctival phagocytic cells. Allogeneic pIC-activated conjunctival MSC injections were well tolerated. GFP-labeled tracking identified MSC components phagocytosed by immune cells subconjunctivally. This preliminary safety and tracking information is critical towards advancing immune conditioned cellular therapies to clinical trials in horses.

A Bayesian network-based model for risk modeling and scenario deduction of collision accidents of inland intelligent ships

Safety is an important premise and foundation for the operation of intelligent ships. This paper introduces a novel scenario analysis framework that employs disaster system theory to produce more comprehensive results for identifying scenario elements and calculating collision risks for inland intelligent ships. The framework is utilized to investigate the collision accident risk evolution mechanism. This process is incorporated into Bayesian Network (BN) modeling for ship collisions on inland rivers. By comparing the change in occurrence probability and consequence severity of risk factors for inland ship collision accidents with and without selected intelligent technologies, the collision risk of intelligent ships is quantified. The results indicate that the application of intelligent technologies, such as ship speed optimization and situational awareness, can reduce the occurrence probability of collision accidents and mitigate the severity of their consequences. Moreover, it has been discovered that such intelligent technologies have a greater impact on accidents with severe consequences than those with minor consequences. This research provides a framework for the preliminary safety evaluation of inland intelligent ships. It is of great significance to accelerate the improvement of navigational risk prevention and response-ability of inland intelligent ships in the future.

Pharmacokinetic study and preliminary evaluation of safety and efficacy of the recombinant human monoclonal antibodies against rabies virus (rhRIG) in Chinese healthy population: A randomized, single-blinded, placebo-controlled phase Ia clinical trial

BackgroundPassive immune agents play an important role in the prevention of rabies following exposure. This trial investigated the safety, tolerability, and pharmacokinetics of recombinant human monoclonal antibodies against rabies virus (rhRIG). MethodThis clinical trial was conducted on healthy Chinese adults. The subjects were enrolled into three dosage groups. The safety of the drug was assessed, and the blood concentration of the monoclonal antibody (NM57) and the neutralizing antibody levels were measured. ResultsThe rhRIG presented favorable safety and tolerability.The subjects in the low(10 IU/kg), medium(20 IU/kg), and high(40 IU/kg) dosage groups reported the Cmax of 87.15 ± 18.86, 210.92 ± 77.5, and 394.11 ± 134.98 ng/ml, respectively, and the AUC0-t of 2115.8 ± 791.3, 6064 ± 1890, and 10735.6 ± 4090 ng/mL*d, and the T1/2 z of 15.82 ± 3.03, 15.96 ± 2.37, and 14.34 ± 3.84 d. The Tmax of neutralizing antibody levels in the serum was about 10d (5-14d). The detectable rate of neutralizing antibody levels in the medium dosage group was close to that in the high dosage group and higher than that in the low dosage group. ConclusionsThe 10–40IU/kg rhRIG was well tolerated by the participants. The results of the pharmacokinetic analysis were consistent with the characteristics of linear elimination. The neutralizing antibody levels was positively correlated with the dosage of rhRIG.

An at-home, virtually administered graded exertion protocol for use in concussion management: Preliminary evaluation of safety and feasibility in healthy youth and children with subacute concussion.

Graded exertion testing (GXT) is an important tool for concussion management, as it is used to personalize post-concussion exercise prescription and return athletes to sport. However, most GXT requires expensive equipment and in-person supervision. Our objective was to assess the safety and feasibility of the Montreal Virtual Exertion (MOVE) protocol, a no-equipment, virtually compatible GXT, in healthy children and children with subacute concussion. Cross-sectional. The MOVE protocol consists of seven stages of bodyweight and plyometric exercises performed for 60 seconds each. Twenty healthy (i.e., non-concussed) children completed the MOVE protocol virtually over Zoom Enterprise. Next, thirty children with subacute concussion (median: 31.5 days post-injury) were randomized to the MOVE protocol or Buffalo Concussion Treadmill Test (BCTT), which increases the incline or speed of the treadmill every minute until maximum exertion. Out of an abundance of caution, all concussed participants completed the MOVE protocol in an in-person clinical space. However, the test evaluator was stationed in a different room within the clinic and administered the MOVE protocol using Zoom Enterprise software to mimic telehealth conditions. Safety and feasibility outcomes were recorded throughout GXT, including heart rate, rate of perceived exertion (RPE), and symptom outcomes. No adverse events were recorded, and all feasibility criteria were successfully met in healthy youth and youth with concussion. Among concussed youth, increases in heart rate (MOVE: 82.4  17.9 bpm, BCTT: 72.1  23.0 bpm; t(28)=1.36, p=0.18), RPE (MOVE: 5.87  1.92, BCTT: 5.07  2.34, t(28)=1.02, p=0.32), and overall symptom presentation were similar between the MOVE and BCTT protocols. The MOVE protocol is a safe and feasible GXT that in healthy youth and youth with subacute concussion. Future studies should assess the fully virtual administration of the MOVE in children with concussion, MOVE protocol tolerability in children with acute concussion, and whether the MOVE protocol can be used to guide individualized exercise prescription.

Thoracic paravertebral block vs mid-point transverse process to pleura block in thoracic surgery. Preliminary evaluation of effectiveness and safety

Mid-point Transverse process to Pleura Block (MTPB) is an alternative approach to paravertebral block. We retrospectively assessed hemodynamic stability, inadvertent pleural puncture and antalgic effectiveness of this approach compared with traditional paravertebral block. Moreover, being used to add methylene bleu to local anesthetic solution as a tracer, we compared the diffusion pattern in paravertebral space of the two techniques. We reported retrospectively 40 patients underwent lung lobectomy from July to October 2021 in Monaldi Hospital (Naples, Italy) receiving thoracic paravertebral block (n=20) or mid-point transverse process to pleura block (n=20). The primary outcomes were a 20% mean arterial pressure variation from the base line, chest wall or pleural hematoma and technique failure. Analgesic effectiveness was assessed comparing remifentanil consumption to keep nociception index level &lt; 25 and patient’s VAS in post anesthesia care unit or daycare surgical unit at 60’. Also the methylene blue spread was assessed by a surgeon, blinded to patient’s name reviewing surgery videorecord of every cases. MAP decrease &gt; 20% was greater in the TPVB. No significative differences were reported for pleural punctures, chest wall hematomas or failure of the technique in the two groups. Methylene blue was evident in all patients of the TPVB, while it was not visible in any of the patients of the MTPB group. Remifentanil consumption, nociception index level and postoperative 60’ VAS were greater in the MTPB.Antalgic power and local anesthetic spread of TPVB seems to be superior to the MTPB, even if the safety profile of the latter seems to be better. Due to the retrospective nature of the study and the small number of cases in the sample, further studies are needed.

Abstract LB324: BB3008, a potent and selective small molecular HPK1 inhibitor effective in multiple syngeneic tumor models

Abstract Introduction: Hematopoietic progenitor kinase 1 (HPK1) is a serine/threonine kinase in the MAP4K family predominantly expressed in immune cells, and has been identified as a negative regulator of T cell and B cell receptor signaling and dendritic cell function. Inhibition of the kinase activity of HPK1 results in the activation of exhausted T cell in tumor microenvironment and enhanced anti-tumor immune response. Observation of HPK1 overexpression in various tumor tissues further indicates the kinase as a novel target for immuno-oncology therapy. Here we report BB3008 as a small molecular HPK1 inhibitor. Methods: BB3008 was developed through structure-based drug design, and optimized by SAR analysis and medicinal chemistry iteration. Biochemical assays and cell-base functional assays were applied in compound evaluation. Pharmacokinetic (PK) and preliminary toxicity studies were performed with mice and dogs. Syngeneic tumor models in mice were conducted to demonstrate the in vivo efficacy of BB3008 as a single agent and in combination with mouse PD-1 antibody. Results: BB3008 shows sub-nanomolar HPK1 potency, with good selectivity against kinases in the MAP4K family and other TCR signaling-related kinases, such as FYN and ZAP70. Inhibition of HPK1 by BB3008 strongly suppresses the phosphorylation at Ser376 of the downstream biomarker protein SLP-76, with IC50 at 30 nM. With the stimulation of CD3/CD28, boost of IL-2 production was observed in Jurkat cells, purified human T cells, and human peripheral blood mononuclear cells upon treatment of BB3008 at sub-micromolar concentration. BB3008 also enhanced the killing activity of T cell against EL4 lymphoma cell in vitro in a concentration-dependent manner. PK profile of BB3008 showed good bioavailability (&amp;gt;80%) with a nearly linear dose-dependent exposure. The compound had no significant suppression on various normal cell lines proliferation, and had no hERG liability. In preliminary safety evaluation, no abnormal clinical symptoms and serum/blood tests were observed in mice and dogs dosed up to 1000mpk and 150mpk for 14 days, respectively. BB3008 showed significant tumor-growth inhibition rates in CT26, Hepa 1-6 and 4T1 mouse models as single agent, and in MC-38 mouse model in combination with mouse PD-1 antibody. Conclusion: As a potent, selective and oral available small molecule HPK1 inhibitor, BB3008 shows good safety tolerance in preclinical animals, and promising efficacy in multiple solid tumor models, both as single agent or in combination with immune checkpoint blockade. The IND-enabling study of BB3008 is in progress. Citation Format: Gongping Duan, Min Li, Xingmin Zhang. BB3008, a potent and selective small molecular HPK1 inhibitor effective in multiple syngeneic tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB324.

Abstract 2933: Novel CD123xCD3 bispecific IgM antibody, IGM-2537, potently induces T-cell mediated cytotoxicity of acute myeloid leukemia cells with minimal cytokine release

Abstract Novel therapeutics are needed for the effective treatment of acute myeloid leukemia (AML), as standard chemotherapeutics are poorly tolerated and ~50% of patients relapse primarily due to the incomplete elimination of leukemia stem cells (LSCs). CD123, the IL-3Rα chain, is highly expressed on leukemic blasts, LSCs, and is further increased in patients with poor prognostic factors. Bispecific T-cell engager (TCE) antibodies and CAR-T cells targeting CD123 have shown promising clinical efficacy in AML patients, but cytokine release syndrome limits their therapeutic window and remains a major safety concern. IGM-2537 is a novel pentameric IgM bispecific TCE antibody engineered with ten anti-CD123 binding sites, and an anti-CD3ε single chain Fv domain fused to the joining chain to engage T-cells. Here, we report the functional characterization of IGM-2537 using in vitro, ex vivo and in vivo anti-tumor efficacy studies with preliminary safety evaluation of this novel class of IgM TCE. IGM-2537 bound with high selectivity, affinity and avidity to CD123 through an epitope distinct from IL-3. In vitro, IGM-2537 co-engaged with both CD123 and CD3 to induce potent T-cell activation and T-cell mediated cytotoxicity of AML cell lines. Though IGM-2537 demonstrated comparable maximal killing activity to a comparator IgG TCE, IGM-2537 demonstrated minimal cytokine release. In ex vivo patient-derived AML or normal bone marrow colony formation assays, IGM-2537 eliminated AML colony forming cells at physiologically relevant effector/target (E/T) ratios but spared normal progenitors. In addition, IGM-2537 potently depleted CD123+ basophils and plasmacytoid dendritic cells (pDC) in normal human peripheral blood mononuclear cells. In vivo, IGM-2537 completely inhibited tumor growth in the AML xenograft tumor model, MV4-11 in humanized NSGdKO mice at doses as low as 1 mg/kg. To further evaluate the potency and safety of the CD123xCD3 IgM bispecific TCE format in vivo, a cynomolgus cross-reactive CD123xCD3 IgM bispecific TCE was evaluated for tolerability and pharmacodynamic responses in cynomolgus monkeys. All animals tolerated this bispecific IgM TCE well at doses up to 10 mg/kg (the maximal dose level evaluated), a dose 100-fold greater than published doses of a comparator IgG TCE. Complete depletion of CD123+ basophils and substantial reductions of pDCs were seen in blood and bone marrow with minimal to no cytokine induction. In summary, IGM-2537 demonstrated potent in vitro and in vivo T-cell mediated cytotoxicity of AML cell lines with minimal cytokine induction. These preclinical data further support the clinical development of IGM-2537 for the treatment of AML and more broadly substantiate the use of IgM antibodies as a framework for TCEs to provide an improved therapeutic window for T-cell redirected therapeutics. Citation Format: Gene Li, Ling Wang, Poonam Yakkundi, Paul Hinton, Deepal Pandya, Keerthana Sekar, Rodine Rosete, Zhongde Ye, Nardeen Hanna, Maya F. Kotturi, Liz Bogaert, Jiyoung Hong, Christina Tsai, Thomas Manley, Bruce A. Keyt, Angus M. Sinclair, Liqin Liu. Novel CD123xCD3 bispecific IgM antibody, IGM-2537, potently induces T-cell mediated cytotoxicity of acute myeloid leukemia cells with minimal cytokine release [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2933.

Bestatin analogs-4-quinolinone hybrids as antileishmanial hits: Design, repurposing rational, synthesis, in vitro and in silico studies

Amongst different forms of leishmaniasis, visceral leishmaniasis caused by L. donovani is highly mortal. Identification of new hit compounds might afford new starting points to develop novel therapeutics. In this lieu, a rationally designed small library of bestatin analogs-4-quinolone hybrids were prepared and evaluated. Analysis of SAR unveiled distinct profiles for hybrids type 1 and type 2, which might arise from their different molecular targets. Amongst type 1 bestatin analog-4-quinolone hybrids, hybrid 1e was identified as potential hit inhibiting growth of L. donovani promastigotes by 91 and 53% at 50 and 25 μM concentrations, respectively. Meanwhile, hybrid 2j was identified amongst type 2 bestatin analog-4-quinolone hybrids as potential hit compound inhibiting growth of L. donovani promastigotes by 50 and 38% at 50 and 25 μM concentrations, respectively. Preliminary safety evaluation of the promising hit compounds showed that they are 50–100 folds safer against human derived monocytic THP-1 cells relative to the drug erufosine. In silico study was conducted to predict the possible binding of hybrid 1e with methionine aminopeptidases 1 and 2 of L. donovani. Molecular dynamic simulations verified the predicted binding modes and provide more in depth understanding of the impact of hybrid 1e on LdMetAP-1 and LdMetAP-2.

Radiological safety evaluation of a recycling facility for dismantled concrete waste

When decommissioning a nuclear power plant, a large amount of radioactive waste will be generated. It must be disposed of in a safe and efficient way. Approximately 50% of radioactive waste is concrete waste. The bio-shield is expected to be highly activated compared to concrete structures because of its continuous radiation by neutron flux. Therefore, the spatial dose distribution of the recycling facility and the acquired doses of workers must be determined when recycling or disposing of bio-shield concrete. This study constitutes a preliminary evaluation of radiation safety for a recycling facility for dismantled concrete structures. The radiation dose was simulated using VISIPLAN code to simulate the spatial dose distribution and the external dose of radiation workers. The internal dose of radiation workers was also calculated. The most significant factor in the dose assessment of concrete waste recycling is the radioactivity value of the waste. Thus, this radioactivity value used by cutting the bio-shield is important. It is classified into three cases as follows: the section of the bio-shield that was exposed to the highest radiation levels (case 1), a vertical cut through the section closest to the nuclear reactor (case 2), and where the concrete waste exceeded the allowable concentration for the regulatory clearance (case 3). Case 3 includes parts of cases 1 and 2, as wastes with concentrations higher than the allowable limit for regulatory clearance are classified regardless of the cutting method. The annual effective doses of a worker were 4.40E+01 mSv, 9.07E+00 mSv, and 5.19E+00 mSv for cases 1, 2, and 3, respectively. We determined that the stipulated safety thresholds of 100 mSv over 5 years and no more than 50 mSv in any single year were only exceeded (specifically, the 5-year limit) for case 1, under the assumptions made in our study. We demonstrated that lead shielding is highly effective in reducing doses. Shielding corresponding to thickness of 10 mm and 20 mm, respectively, produced reductions in dose rates of 43.5% and 65.8%, respectively. The workers' annual doses in all cases except case 1 with lead 10 mm when adding lead shielding did not exceed the stipulated annual dose limit of 20 mSv. This dose evaluation of the work processes of a concrete recycling facility is expected to be applied to the actual management of workers’ radiation safety.

Preliminary Safety and Efficacy Evaluation of IMM0306, a CD47 and CD20 Bispecific Monoclonal Antibody-Trap (mAb-Trap), from an Ongoing Phase I Dose-Escalation Study in Patients with Relapsed or Refractory B-Cell Non- Hodgkin's Lymphoma (R/R B-NHL)

Background: IMM0306, a recombinant bispecific mAb-Trap targeting CD20 and CD47, has higher affinity for CD20 than for CD47, thus enabling its preferential and simultaneous binding to CD20 and CD47 on malignant B cells rather than to CD47-postive normal tissues. Here, we present preliminary safety and efficacy data from an ongoing dose-escalation study in patients with relapsed or refractory B-cell non-Hodgkin's lymphoma (R/R B-NHL) (CTR20192612). Methods: IMM0306-I is a first-in-human, phase I dose escalation and dose expansion study of IMM0306. 7 dose levels (0.04, 0.1, 0.25, 0.5, 0.8, 1.2, 1.6 mg/kg) were to be evaluated in the dose escalation phase of the study following a modified 3+3 escalation schema. After single-dose period (2 weeks), IMM0306 was administered intravenously once per week (QW) until disease progression or intolerable toxicity. Dose-limiting toxicity (DLT) was evaluated in the 28 days post the initial 2-week single dose period. Safety was evaluated per CTCAE 5.0; tumor assessments performed once every 8 weeks, and pharmacokinetics (PK) assessed after single and multiple doses of IMM0306 administration. Results: As of Jul 13th, 2022, 26 patients were treated with seven dose levels of IMM0306. 10 (39%) pts received 3 or more prior lines of therapy; all pts received prior anti-CD20 therapy. No DLTs have been observed by the date of data cut-off and the maximum tolerated dose (MTD) not reached. IMM0306 was generally well tolerated. The most frequent (>20%) treatment-related adverse events (TRAEs) were lymphocyte decrease (65.4%), White Blood Cell (WBC) decrease (61.5%), neutropenia (50.0%), anemia (50.0%), infusion- related reactions (IRR) (38.5%), platelet (PLT) count decrease (30.8%), most of them are Grade1-2. The most common G3 or above TRAE were lymphocyte decrease (65.4%), WBC decrease (23.1%), absolute neutrophil count (ANC) decrease (15.4%), anemia (11.5%), PLT count decrease (7.7%), respectively. Two treatment related SAE patients are G4 PLT decrease and chest pain respectively. These two patients discontinued the study treatment after the event resolved (note: this subject reported G4 PLT decrease at the dose of 1.6 mg/kg at C1D30). Among total 23 evaluable patients, one follicular lymphoma patient achieved complete response (CR) at the dose level of 1.2 mg/kg. 2 partial response (PR) at 0.8 and 1.2 mg/kg dose level (one is for marginal zone lymphoma, the other is for follicular lymphoma) respectively. Six patients reached stable disease (SD). PK data has yet to be collected. Conclusions: These data indicate that IMM0306 is well-tolerated at the dose levels evaluated. and preliminary anti-tumor efficacy result observed support continuous development of IMM0306 in patients with R/R B-NHL. The phase 1/2 is ongoing.

Discovery and Preclinical Study of Novel BTK Degrader HZ-Q1060

Introduction: Bruton's tyrosine kinase （BTK） is a clinical validated drug target for therapy of B cell lymphoma and autoimmune diseases. However, same as other kinase inhibitors, acquired drug resistance, which is the result of BTK C481S mutation, has been observed in patients. So reversible BTK inhibitors were developed to overcome acquired drug resistance of covalent BTK inhibitor. While, in order to have enough efficacy, reversible BTK inhibitors required much higher drug exposure which may lead to higher risk of adverse effect. Hence, it is urgent to develop new technique to overcome drug resistance caused by BTK C481S mutation. Proteolysis-targeting chimeras(PROTAC) is promising new strategy for drug development. PROTAC can achieve sustained target degradation in a catalytical mechanism. So comparing with traditional small molecule inhibitor, PROTAC drug has great advantage in solving acquired drug resistance. Therefore, with the aim of solving acquired drug resistance of 1st generation of BTK inhibitor, we identified a novel BTK-PROTAC HZ-Q1060 based on our in-house DaTProD® platform which is setting up to promote PROTAC development. After a series of evaluations, HZ-Q1060 has been validated as a promising pre-clinical candidate for further clinical development. Results:HZ-Q1060 can effectively degrade BTK in cancer cell lines and human PBMC, and the half degradation concentration (DC50) in Mino is less than 0.5nM. At the same time, HZ-Q1060 can also induce the degradation of intracellular mutant BTK-C481S protein.HZ-Q1060 has not observed any inhibition and degradation effect on other kinases except BTK, especially kinases that may be related to potential side effects by inhibiting, such as EGFR and TEC.In vitro cancer cell growth inhibition assay, it completely suppressed the cell proliferation of a panel of lymphoma cell lines such as Mino(IC50=0.11nM) and OCI-ly10(IC50=0.28nM).In the PK/PD experiment in mice, a single oral administration of medium dose can degrade more than 80% of BTK within 6 hours, and the degradation effect can be maintained until 24 hours after administration.In the subcutaneous transplanted tumor model of OCI-ly10 mice, the tumor growth was completely inhibited by 3mg/kg of HZ-Q1060 administered orally every day for 14 days, which was equivalent to that of ibrutinib 10mg/kg.In the preclinical pharmacokinetic studies, HZ-Q1060 showed excellent pharmacokinetic properties in different species of animals, included mice, rats and beagle dogs, with long T1/2 and high dose-normalized AUC.HZ-Q1060 has no significant CYPs inhibitory effect at 10 μM, so it has a low risk of DDI in the future combination therapy.HZ-Q1060 has no inhibitory effect on hERG channel, indicating the compound has a low potential to induce QT prolongation. HZ-Q1060 also shows good safety and tolerance in the preliminary safety evaluation of animals. Conclusion: In summary, HZ-Q1060 is a potent and selective degrader of BTK with good ADME properties and in vivo efficacy, which makes it suitable for further evaluation in clinical development.

Safety assessment of existing RC bridge piers subjected to vehicle impact

Impact accidents between bridges and vehicles have great randomness, and the threat of various vehicles to bridge structures is also different. In this case, the vehicle impact shall be regarded as a risk event from the perspective of economic and safety requirements, but current studies mainly focus on the deterministic analysis of bridges subjected to vehicle impact. For this reason, this work aims to establish a probability-based safety assessment method for existing reinforced concrete (RC) bridge piers against potential vehicle impact based on the reliability theory. Firstly, a simplified approach is developed for the preliminary impact hazard analysis of RC piers using the probability method, in which the vehicle type and impact probability are considered. Then, the shear and flexural responses of RC piers under vehicle impact are numerically investigated, and the corresponding limit state functions are determined. Furthermore, a procedure to assess the safety level of RC piers is proposed, including preliminary hazard analysis, reliability analysis, and safety evaluation. Finally, a case example of a bridge pier against vehicle impact is given to demonstrate the proposed safety assessment procedure. This method could provide a reference and basis for the anti-impact decision-making of bridge structures in their service period.