Menarini Group (Italy)

The present study investigated the combined effects of zofenoprilat (ZOFE) and nebivolol (NEBI) on endothelial function, focusing on their anti-inflammatory and antioxidant properties. The purpose was to evaluate whether these drugs, commonly used in clinical practice, offer a synergistic therapeutic strategy for managing hypertension and protecting vascular health. ZOFE, an ACE inhibitor, demonstrated significant anti-inflammatory activities by reducing inflammatory cytokines, thereby mitigating vascular inflammation, a key factor in hypertension and atherosclerosis. NEBI, a third-generation beta-blocker, exhibited strong antioxidant effects by enhancing nitric oxide (NO) levels, crucial for maintaining endothelial function and reducing oxidative stress. The potential effect of ZOFE and NEBI treatment was evaluated using human umbilical vein endothelial cells (HUVEC) as a model. Specifically, cells were challenged with tumor necrosis factor-α (TNF-α) to induce endothelial dysfunction. Subsequently, cell viability, NO production, protein levels of superoxide dismutase (SOD) and catalase (CAT), enzymatic activity of SOD and CAT, intracellular levels of glutathione (GSH), inflammatory status, and levels of interleukin-6 (IL-6) monocyte chemoattractant protein-1 (MCP-1), macrophage inhibitory cytokine-1 (MIC-1), and the active form of nuclear factor kappa B (p-NFκB), were analyzed. Our results showed that NEBI significantly counteracted oxidative stress, increasing the main antioxidant defenses (SOD, CAT, and GSH). The combination of ZOFE and NEBI resulted in a potentiated effect, enhancing both anti-inflammatory and antioxidant activities. This dual mechanism of action provides a comprehensive approach to protecting endothelial cells and improving vascular function. The combined therapy not only lowered blood pressure more effectively but also offered greater protection against endothelial damage compared to monotherapy with either drug alone. These findings suggest that the combination of ZOFE and NEBI could be particularly beneficial for patients with hypertension, especially those with coexisting inflammatory and oxidative stress-related conditions. This combination therapy, by addressing multiple pathogenic pathways simultaneously, could potentially be beneficial in patients with cardiovascular risk conditions. In conclusion, the combination of ZOFE and NEBI offers a potentially promising therapeutic approach for managing hypertension and protecting vascular health, aiming at improving clinical outcomes for patients with cardiovascular diseases. Not applicable.

Patients with myelofibrosis (MF) who are resistant to or relapse after Janus kinase inhibitor (JAKi) therapy have limited treatment options and typically poor prognoses. CD123 is overexpressed in various myeloid malignancies, including MF. Tagraxofusp is a first-in-class CD123-targeted therapy, and the only drug approved globally for the rare myeloid malignancy blastic plasmacytoid dendritic cell neoplasm. We conducted a phase 1/2 trial to determine optimal dosing, and evaluate safety and efficacy of tagraxofusp monotherapy in treatment-naïve (n = 5) MF and patients with MF resistant/refractory to JAKi (n = 25) and not eligible for stem cell transplant. There were no dose-limiting toxicities. The recommended phase 2 dose of tagraxofusp was 12 μg/kg per day for 3 consecutive days per cycle. In the safety population (n = 36) treated at 12 μg/kg per day, the most frequent grade ≥3 treatment-emergent adverse events were thrombocytopenia (19%), anemia (22%), and dyspnea (11%). Capillary leak syndrome occurred in 11% of patients, all during cycle 1 with resolution in all patients. Thirty patients treated at 12 μg/kg per day were efficacy evaluable. Of 18 (n = 2 treatment-naïve, n = 16 relapsed/refractory) patients with baseline splenomegaly, 2 relapsed/refractory patients had spleen volume reduction ≥35%. In relapsed/refractory patients, 40% had total symptom score (TSS) ≥50%, and median overall survival (OS) was 19.3 months. In patients who were treatment naïve, 40% had TSS ≥50%, and median OS was 26.6 months. In this trial, tagraxofusp monotherapy in MF was well tolerated, without cumulative myelotoxicity, and with symptom score improvements, warranting further investigation in combination therapy. This trial was registered at www.clincaltrials.gov as #NCT02268253.

MEN1703 is a first-in-class, oral, Type I dual PIM/FMS-like tyrosine kinase 3 inhibitor (FLT3i) investigated in a Phase I/II DIAMOND-01 trial in patients with acute myeloid leukaemia (AML). Gilteritinib is a highly potent and selective oral FLT3i approved for the treatment of relapsed/refractory AML with FLT3 mutations. Although gilteritinib showed strong single-agent activity in FLT3-mutated AML, the development of gilteritinib resistance limits response durability, indicating the importance of novel combination strategies to improve disease outcome. PIM kinases govern FLT3-ITD signalling and increased PIM kinase expression is found in samples from AML patients relapsing on FLT3i. Here, we report that the simultaneous inhibition of PIM and FLT3, through the combination of MEN1703 and gilteritinib, can consistently improve the invitro/in vivo antitumor activity over the single agents, demonstrating the benefit of this combination. Moreover, we demonstrate that resistance to gilteritinib can be circumvented by combining MEN1703 with gilteritinib. MEN1703 interferes with FLT3 upregulation, Mcl-1 overexpression and PIM kinase signalling, which are all involved in FLT3i resistance. We also show that MEN1703 downregulates stromal cytokines that promote cytokine-mediated resistance of AML blast cells to FLT3 inhibition. These results demonstrate the importance of the combination approach to overcome microenvironment-mediated resistance to FLT3 inhibitors.

The integration of magnetic resonance (MR) imaging and linear accelerators into hybrid treatment systems has made MR-guided radiation therapy a clinical reality. This work aims to evaluate the influence of the Electron Return Effect (ERE) on the dose distributions. This study was conducted using MRIdian (ViewRay, Cleveland, Ohio) system. Monte-Carlo simulations (MCs) and experimental measurements with EBT3 Gafchromic films were performed to investigate the dose distribution in a slab water phantom with and without a 2-cm air gap. Plus, MCs took into account different field sizes and a lung gap. A gamma analysis compared calculated versus measured dose distributions. The MCs have shown an increase of the ERE with the radiation field size both in Percent Depth Dose (PDD) and crossline direction. As concerns to the PDD direction, the smallest field for which there was a significant dose accumulation was 4.15×4.15 cm2both for air-gap (13.5%) and lung-gap (3.3%). The largest field for which there was a significant dose accumulation was 24.07×24.07 cm2both for air-gap (39.7%) and lung-gap (4.9%). Instead for the crossline direction, the smallest field for which there was a significant dose accumulation was 2.49×2.49 cm2both for air-gap (8.6% ) and lung-gap (0.5%). The largest field for which there was a significant dose accumulation was 24.07×24.07 cm2both for air-gap (46.2%) and lung-gap (4.2%). PDD and crossline profiles showed good agreement with a gamma-passing rate higher than 91.15% for 2%/2 mm. The ERE can be adequately calculated by MC dose calculation platform available in the MRIdian Treatment Planning System. The MCs show an increase of the ERE directly proportional with the radiation field size. Good agreement was observed between the experimental measurements and calculated dose distributions.

Pathogenic microdeletions/duplications (pCNVs) account for a significant perinatal morbidity/mortality and have a 1/100-200 prevalence in the general prenatal population. Cell-free DNA non-invasive testing (cfNIPT) shows significant limitations for comprehensive profiling of pCNVs <7Mb in size. A proof-of-concept study demonstrated the feasibility of a novel cell-based NIPT (cbNIPT) for circulating extravillous trophoblasts (cEVTs) isolation and single cell sequencing for fetal CNV detection down to ~800kb. The scope of the study is to demonstrate cbNIPT scientific validity in a large blinded prospective performance study comparing the fetal genomic result with that of cEVTs screening. 1388 high-risk pregnancies were enrolled to obtain a statistically significant pCNV/aneuploidies number. Blood was collected from pregnant women at 11-22+6 wg before invasive diagnostic procedure and processed with proprietary laboratory workflow not optimised for recovery rate. 1070 had CMA ± karyotype and the remainders with karyotype only. Screen positive rate for aneuploidies and pCNV was 18.9% and 11.2%, respectively. Totally 203 pCNVs ≥ 1Mb were detected and 84.2% of them (171/203) were 1-7 Mb in size. Of note, pCNVs < 1 Mb were also detected. Examples of pCNV are: 22q11.2del (7 spanning 600Kb-3Mb), 22q11dup (1 of 3Mb), 2q37del (2 of 2.1Mb and 3.9Mb), 8q24.3del (2 of 600kb), 7q22.3del (1 of 800Kb), 1q21.1del (1 of 800kb), Wolf-Hirschhorn Sy (1 of 2.9Mb) and 1 Williams Beuren Sy (1 of 1Mb). At the time of submission, the clinical database was locked and confirmatory clinical data analysis was ongoing. Analysis of cEVTs from maternal blood offers the detection of genome wide pCNV < 1 Mb, below the resolution of cell-free DNA screening. In the general pregnancy population, this would allow the opportunity for a significant reduction of residual risk for pCNV at early gestational weeks. CbNIPT clinical performance data will be provided together with an estimated rate of detection compared to cfNIPT.

Abstract Background: ER+/HER2- breast cancer (BC) is the most prevalent BC subtype. Endocrine therapy (ET) is the mainstay of treatment; however, development of resistance to this therapeutic approach is common in the metastatic setting. After progression on first-line treatment, both ET monotherapy and combination therapy options are available. Combinations such as everolimus + exemestane and alpelisib + fulvestrant can be associated with higher efficacy, but also significant toxicity with discontinuation rates around 25%. Also, for second-line treatment, pts who did not receive prior treatment with a CDK4/6i, the combination of fulvestrant with a CDK4/6i (palbociclib, abemaciclib, and ribociclib) are among the recommended treatment options for this pt population (NCCN, 2022). However, fulvestrant has low bioavailability and an IM injection burden. The crosstalk between the three pathways ER, PI3K/AKT/mTOR and cyclin-dependent kinases (CDK4/6) provide a rationale for the combination of elacestrant, a next-generation oral SERD, with inhibitors of these pathways that are natural targets to overcome endocrine resistance. The EMERALD trial reported significantly prolonged progression-free survival (PFS) with elacestrant vs SOC ET in pts with ER+/HER2− ESR1 mutated (ESR1-mut) mBC following disease progression on prior ET (Bidard, 2022). Elacestrant was well tolerated with a manageable safety profile. Most adverse events, including nausea, were low-grade and consistent with other endocrine therapies. The replacement of the fulvestrant backbone with elacestrant in combination with targeted agents enabling oral-oral combinations is of therapeutic interest. Methods: ELEVATE (NCT05563220) is a phase 1b/2 clinical trial that will evaluate the safety and efficacy of elacestrant combined with alpelisib, everolimus, palbociclib, abemaciclib, and ribociclib. Eligible pts must be adults with confirmed ER+/HER2− advanced or mBC, &amp;gt; 1 measurable lesion as per RECIST v1.1, and adequate bone marrow or organ function. The main objectives for Phase 1b are to determine the RP2D of elacestrant combined with other targeted agents. The RP2D of elacestrant and abemaciclib will be determined in the ELECTRA study (NCT05386108). The Phase 2 portion will evaluate the efficacy of elacestrant combined with other study drugs for PFS; secondary objectives include PK of the various combinations, ORR, DoR, CBR, PFS, and OS. Here we report the initial dosing from the Phase 1b portion. Results: As of June 2023, 11 pts were enrolled in the phase 1b portion of the trial, cohort 1 for all combinations: 5 pts in the elacestrant and everolimus (300 mg elacestrant QD/everolimus 5 mg QD) and 3 pts in the elacestrant and alpelisib (300 mg elacestrant QD/alpelisib 250 mg QD) and 2 patients for palbociclib and 1 patient in the ribociclib combinations. Further patient accrual is expected in phase 1b cohorts of various combinations during the second half of 2023; both safety and PK data will be provided. Conclusion: In the Phase 1b portion of ELEVATE, elacestrant is being combined with various targeted agents enabling oral-oral combinations to determine a RP2D. Recruitment is actively ongoing; safety and PK results will be presented at the meeting from the completed phase 1b cohorts. Citation Format: Hope Rugo, Aditya Bardia, Javier Cortés, Giuseppe Curigliano, Erika Hamilton, Sara Hurvitz, Sibylle Loibl, Sara Tolaney, Virginia Kaklamani, Giulia Tonini, Shannon Matheny, Kathy Puvana Theall, Joyce O'Shaughnessy. ELEVATE: A phase 1b/2, open-label, umbrella study evaluating elacestrant in various combinations in patients (pts) with estrogen receptor-positive (ER+), HER2-negative (HER2-) locally advanced or metastatic breast cancer (mBC) [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-05-04.

Abstract Myelofibrosis (MF) is a pathological chronic bone marrow (BM) condition characterized by abnormal growth of erythroid and megakaryocytic precursors and deposition of fibrotic tissue. The JAK/STAT pathway plays an important role in MF (Cardoso 2015), and multiple JAK1/2 inhibitors (JAKis) are approved for MF treatment. Despite initially limiting disease symptoms, JAKis often fail due to persistent JAK2 activation and survival of MF-driving cells, hence new treatment strategies are highly needed. PIM kinases also contribute to MF pathogenesis. PIM1 is highly overexpressed in granulocytes, peripheral blood mononuclear cells (PBMCs), and BM of patients with MF. PIM1/2 expression is dependent upon JAK2 signaling in myeloproliferative neoplasms (MPNs). Hence, PIM inhibitors represent potential therapeutic targets for MPNs and have been evaluated in several preclinical and clinical studies (Rampal 2021). MEN1703/SEL24 (MEN) is an oral, first-in-class, dual PIM/FLT3 kinase inhibitor in development for hematologic malignancies. This study aims to investigate the efficacy of MEN alone and in combination with the JAKi ruxolitinib (RUX) in preclinical MF models and to elucidate the underlying signaling pathways. MF cell lines (JAK2V617F and JAK2 wild type) were used in MTS assays to assess the in vitro cytotoxicity of MEN alone and in combination with RUX, compared with the PIM inhibitor TP-3654 (Dutta 2022). Cell cycle distribution and signaling pathways were analyzed via propidium iodide/FACS analysis and immunocapillary electrophoresis, respectively, upon treatment with the single agents and MEN + RUX. In vivo efficacy was assessed in a murine xenograft model treated with MEN +/- RUX. In vitro studies showed single agent MEN activity with an IC50 ranging from 0.3 to 2.3 µM. Moreover, a synergistic drug interaction was found with MEN + RUX in all tested cell lines (Combination Index [CI] range 0.41-0.80). MEN presented cytotoxic and cytostatic capacity, inducing apoptosis and arresting a proportion of cells in G2/M phase. This effect was maintained in combination with RUX. Signaling pathway analysis showed that MEN inhibits phosphorylation of STAT proteins and decreases Mcl1 protein expression. These effects were enhanced with MEN + RUX. Compared with TP-3654, MEN was more effective both as a single agent (IC50 2-3 fold lower than TP-3654) and in combination with RUX (mean CI of 0.77-0.80 compared to 0.79-0.98 with TP-3654). MEN showed anti-tumoral efficacy in MF preclinical models, moderately higher than TP-3654, exhibiting in vitro activity at clinically relevant concentrations. Importantly, MEN combined with the standard of care RUX was synergistic, and molecular analyses confirmed the role of PIM downstream target inhibition. Our results support the therapeutic potential and relevance of MEN in MF treatment strategies. Citation Format: Maria Luisa Iannitto, Diego Bisignano, Sonia Zicari, Francesco Belli, Alessio Fiascarelli, Simone Talucci, Giuseppe Merlino, Vittoria Nicolis di Robilant, Clelia Irrissuto, Mario Bigioni, Alessandro Bressan, Daniela Bellarosa, Monica Binaschi. MEN1703/SEL24 exhibits promising antitumoral activity in preclinical models of myelofibrosis both as single agent and combined with ruxolitinib [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 665.

Abstract Purpose. To assess clinical impact and perform cost-consequence analysis of the broadest multiplex PCR panels available for the rapid diagnosis of bloodstream infections (BSI). Methods. Single-center, randomized controlled trial conducted from June 2019 to February 2021 at a French University hospital with an institutional antimicrobial stewardship program. Primary endpoint was the percentage of patients with optimized antimicrobial treatment 12h after transmission of positivity and Gram stain results from the first positive BC. Results. This percentage was significantly higher in the multiplex PCR (mPCR) group (90/105 = 85.7% %, IC95% [77.5 ; 91.8] vs. 68/107 = 63.6%, IC95% [53.7 ; 72.6]; p&lt;10-3) at interim analysis, resulting in the early termination of the study after the inclusion of 309 patients. For patients not optimized at baseline, the median time to obtain an optimized therapy was much shorter in the mPCR group than in the control group (6.9h, IQR [2.9; 17.8] vs 26.4h, IQR [3.4; 47.5]; p=0.001). Early optimization of antibiotic therapy resulted in a non-statistically significant decrease in mortality from 12.4% to 8.8% (p=0.306), with a tendency for a lower median length of stay (18 vs. 20 days; p=0.064) and a non-significant reduction in the average cost per patient of €3,065 (p=0.15). mPCR identified all the bacteria present in 88% of the samples. Conclusion. Despite its higher laboratory cost, the use of multiplex PCR for BSI diagnosis leads to early-optimised therapy, seems cost-effective and could reduce mortality and length of stay. Their impact could probably be improved if implemented 24/7.