Neoadjuvant chemotherapy (CT) followed by radiotherapy (RT) and surgery showed a median survival of 28.7 months in resectable stage IIIB non-small-cell lung cancer (NSCLC) patients (pts). Here, we ...evaluate the impact of concomitant cetuximab to the same neoadjuvant chemo-radiotherapy (CRT) in selected patients (pts) with NSCLC, stage IIIB.
Resectable stage IIIB NSCLC received three cycles of CT (cisplatin 100 mg/m
and docetaxel 85 mg/m
d1, q3w) followed by RT (44 Gy in 22 fractions) with concomitant cetuximab (250 mg/m
, q1w) and subsequent surgery. The primary endpoint was 1-year progression-free survival (PFS).
Sixty-nine pts were included in the trial. Fifty-seven (83%) pts underwent surgery, with complete resection (R0) in 42 (74%) and postoperative 30 day mortality of 3.5%. Responses were: 57% after CT-cetuximab and 64% after CRT-cetuximab. One-year PFS was 50%. Median PFS was 12.0 months (95% CI: 9.0-15.6), median OS was 21.3 months, with a 2- and 3-yr survival of 41% and 30%, respectively.
This is one of the largest prospective phase 2 trial to investigate the role of induction CRT and surgery in resectable stage IIIB disease, and the first adding cetuximab to the neoadjuvant strategy. This trial treatment is feasible with promising response and OS rates, supporting an aggressive approach in selected pts.
BackgroundDose-limiting toxicity (DLT) due to systemic CD40 activation and peripheral target-mediated drug disposition are major challenges in clinical development of CD40 agonists. MP0317, a ...CD40-agonistic DARPin (designed ankyrin repeat protein), is exclusively active in the presence of fibroblast activation protein (FAP) expressed by cancer-associated fibroblasts in the tumor microenvironment (TME). This mode of action enables tumor-localized CD40 activation, while reducing systemic toxicity.MethodsThis ongoing Phase 1, multicenter, open-label, dose-escalation study aims to establish safety/tolerability, pharmacokinetics/pharmacodynamics, and preliminary antitumor activity of MP0317 monotherapy (NCT05098405). The dose-escalation scheme uses an adaptive Bayesian logistic regression model guided by the escalation with overdose control principle to determine the recommended dose. Eligible adult patients with selected advanced solid tumors (based on anticipated FAP expression) are enrolled into 9 sequentially-escalating dose cohorts of MP0317 (0.03–10 mg/kg), administered IV 3-weekly (Q3W) or 1-weekly (Q1W) until disease progression or unacceptable toxicity. The study was approved by the Dutch and French Ethics Boards.ResultsAs of data cut-off (02 May 2023), 36 patients received ≥1 MP0317 dose across 8 cohorts, including 19 women (53%) and 17 men (47%). The median age at enrollment was 63 years (range 35–79) and patients received a median number of 3.5 prior treatments (range 1–13). Colorectal cancer was the most frequent tumor type (11 patients, 31%). One patient experienced a DLT (asymptomatic Grade 3 elevation of alanine and aspartate aminotransferases), at the highest planned dose of MP0317 (10 mg/kg Q3W). Grade 2 infusion-related reaction was the most frequently observed adverse reaction (7 patients, 19%), followed by Grade ≤2 fatigue, nausea and vomiting in 9, 6, and 4 patients, respectively. One patient achieved unconfirmed partial response, and stable disease was observed in 5 patients. Paired tumor biopsies confirmed colocalization of MP0317 with FAP and CD40. MP0317 detection in tumor biopsies was associated with an increase in abundance of antigen-presenting cells (dendritic cells, B cells and plasma cells) and IFNγ signature in the TME. Increases in CXCL10 serum levels post-MP0317 treatment support these findings.ConclusionsThese data of 36 patients, dosed across 8 dose levels (0.03–10 mg/kg, Q3W and Q1W schedules), confirmed a favorable safety profile of MP0317 monotherapy with limited systemic inflammation compared to other CD40 agonists. Analysis of paired tumor biopsies and peripheral biomarkers provided evidence of target occupancy and pharmacodynamic modulation in the TME, consistent with tumor-localized CD40 activation. These data support continued clinical evaluation of MP0317, including combination studies.Trial RegistrationThis study is registered at ClinicalTrials.gov: NCT05098405Ethics ApprovalThe study was approved by the Dutch and French Ethics Boards.
2573 Background: Major challenges of the clinical development of CD40 agonists are toxicity due to systemic CD40 activation and peripheral target-mediated drug disposition. MP0317, a CD40 agonist ...DARPin (designed ankyrin repeat protein), is exclusively activated by binding to fibroblast activation protein (FAP) on cancer-associated fibroblasts. This enables local CD40 activation in the tumor microenvironment (TME) and reduces systemic toxicity. Methods: This Phase 1, multicenter, open-label, dose-escalation study assessed safety/tolerability, pharmacokinetics/pharmacodynamics, and preliminary antitumor activity of MP0317 monotherapy (NCT05098405; data cut-off 15 Jan 2024). Eligible adults with selected advanced solid tumors (based on predicted FAP expression) were enrolled into 9 dose-escalation cohorts of MP0317 0.03–10 mg/kg administered IV 3-weekly (Q3W) or weekly (Q1W) until disease progression or unacceptable toxicity. Blood biomarkers were analyzed by immuno-assays and flow cytometry, and paired tumor biopsies by RNA sequencing and immunofluorescence. Results: Dose-escalation enrolment is complete and 46 patients received ≥1 MP0317 dose, including 24 women (52%) and 22 men (48%). Median age at enrolment was 63 years (range 35–79). Patients received a median of 4 prior treatment lines (range 1–13). Colorectal cancer was the most frequent tumor type (12 patients, 26%). MP0317 maximum tolerated dose was not reached; only one patient experienced a dose-limiting toxicity (asymptomatic Grade 3 alanine and aspartate aminotransferases elevation), at the highest planned dose of MP0317 (10 mg/kg Q3W). Grade ≤2 fatigue was the most frequent adverse reaction (15 patients, 30%), followed by Grade ≤2 infusion-related reaction, nausea and anorexia in 8, 7, and 5 patients, respectively. One patient achieved unconfirmed partial response, and stable disease was observed in 11 patients (24%). Serum PK data showed MP0317’s half-life extended properties and sustained exposure at higher doses. Paired tumor biopsies confirmed the colocalization of MP0317 with FAP and CD40. MP0317 detection in tumor biopsies at doses ≥1.5 mg/kg was associated with an increase in antigen-presenting (dendritic and B cells), plasma and T follicular helper cell abundance, as well as enhanced dendritic cell maturation and IFNγ production in the TME. CXCL10 serum level increases post MP0317 treatment supported these findings. Only minor changes were seen in pro-inflammatory cytokines. Conclusions: MP0317 had a favorable safety profile in 46 patients across all 9 dose-escalation cohorts exploring Q3W and Q1W regimens. Doses ≥1.5 mg/kg showed evidence of pharmacodynamic TME modulation, indicating tumor-localized CD40 activation. The data support further clinical evaluation of MP0317 including combination with complementary anticancer therapies. Clinical trial information: NCT05098405 .
Aims
This study aimed to assess safety, tolerability, pharmacokinetic (PK) and pharmacodynamic (PD) effects of ensovibep, a designed ankyrin repeat protein antiviral being evaluated as a COVID‐19 ...treatment, in healthy volunteers in a first‐in‐human ascending single‐dose study.
Methods
Subjects were dosed intravenously, in a randomized double‐blinded manner, with either ensovibep at 3, 9 or 20 mg/kg or with placebo, and followed until Day 100. PK and safety were assessed throughout the study duration. Immunogenicity and PD via viral neutralization in serum were also assessed.
Results
All adverse events were of mild to moderate severity, and no serious adverse events were observed. One subject who received the 20‐mg/kg dose presented with moderate hypersensitivity vasculitis 3 weeks after infusion, which fully resolved using standard procedures. In most subjects ensovibep showed expected mono‐exponential decline with a half‐life of around 2 weeks. Anti‐drug antibodies were detected in 15 of 17 subjects, with the earliest onset detected on Day 29. Viral neutralization assays on subject serum showed effective viral neutralization over the first 3 weeks following dosing with titre values in a dose dependent manner.
Conclusion
Ensovibep proved safe in this first‐in‐human safety study and exhibited PK and PD parameters consistent with the expected treatment period required for acute COVID‐19 infection.
O6‐methylguanine DNA methyltransferase (MGMT) promoter methylation is a predictive biomarker for benefit from alkylating chemotherapy, specifically temozolomide (TMZ), in glioblastoma, the most ...common malignant intrinsic brain tumor. Glioma‐initiating cells (GIC) with stem‐like properties have been associated with resistance to therapy and progression. We assessed the levels of MGMT mRNA and MGMT protein by real‐time PCR and immunoblot and evaluated the impact of MGMT on TMZ sensitivity in clonogenicity assays in GIC sphere cultures (S) or differentiated adherent monolayer cultures (M). Nuclear factor kappa B (NF‐κB) signaling was assessed by reporter assay and immunoblot. Compared to M cells, S cells expressed higher levels of MGMT. Differentiation of GIC induced by S‐to‐M transition resulted in a gradual loss of MGMT expression and increased TMZ sensitivity. This transcriptional regulation of MGMT was restricted to cell lines without MGMT promoter methylation and was not coupled to any specific neurobasal (NB) stem cell medium supplement or loss of cell adhesion. Expression levels of p50/p65 subunits of NF‐κB, a transcriptional regulator of MGMT, were increased in S cells. Inhibition of NF‐κB by the small molecule inhibitor, BAY 11‐7082, or siRNA‐mediated gene silencing, reduced MGMT levels. In summary, alkylator resistance of S cells is mainly promoted by over‐expression of MGMT which results from increased activity of the NF‐κB pathway in this cell culture model of glioma stem‐like cells.
Read the Editorial Highlight for this article on page 688.
Glioma‐initiating cells are considered to sustain treatment resistance. We demonstrate higher expression of both nuclear factor kappa B (NF‐kB) and O6‐methylguanine DNA methyltransferase (MGMT) in more stem‐like sphere (S) cells compared with monolayer (M) cells of the same cell lines and propose NF‐kB as a regulator of MGMT. We postulate that the higher level of temozolomide (TMZ) resistance in S cells originates from differential regulation of the NF‐kB – MGMT axis.
Read the Editorial Highlight for this article on page 688.
2584
Background: CD40 is a master switch for both innate and adaptive immune systems. The clinical development of CD40 agonists has been hampered by dose-limiting toxicity (DLT) due to systemic CD40 ...activation and peripheral target-mediated drug disposition (TMDD). MP0317 is a CD40-agonistic DARPin (designed ankyrin repeat protein), developed to reduce systemic toxicity. MP0317 is exclusively active in the presence of fibroblast activating protein (FAP) expressed by cancer associated fibroblasts in the tumor microenvironment. This allows reaching serum levels that overcome the TMDD and have the potential to deliver sustained and tumor-localized CD40 activation. Ongoing clinical testing aims at establishing its safety/tolerability profile, PK/PD characteristics, and a recommended dose for combination therapy. Methods: NCT05098405 is a Phase 1, multicenter, open label, dose escalation study followed by a safety expansion part in adult patients with advanced solid tumors. The dose escalation scheme uses an adaptive Bayesian logistic regression model with overdose control. Peripheral blood biomarkers are analyzed by immuno-assays and flow cytometry, and baseline and on-treatment tumor biopsies are characterized by RNA sequencing and immunofluorescence. Eligible patients are enrolled into 9 sequentially escalating dose cohorts of MP0317 (0.03-10 mg/kg), administered IV 3-weekly (Q3W) or 1-weekly (Q1W) until disease progression or unacceptable toxicity. Results: At submission, 23 patients across 6 cohorts completed the study, with no DLT observed. Patient enrolling in higher dose cohorts is ongoing. The most frequent AE was grade 2 infusion related reaction in 5 patients. The 23 patients received ≥2 (range 2-8) doses (range 0.03 – 3 mg/kg) of MP0317 across five Q3W and one Q1W cohorts and completed the 28-day DLT period. PK data confirmed all patients were exposed to MP0317 with broadly dose-dependent Cmax and no sign of accumulation. PD data was derived from patients dosed with 0.03 – 3 mg/kg. Soluble biomarkers (sFAP and sCD40) showed target engagement in periphery with signs of FAP saturation at ≥0.5 mg/kg. Colocalization of MP0317 with FAP and CD40 in the tumor was confirmed in 4 out of 8 evaluable paired tumor biopsies. Whole transcriptome and gene set enrichment analyses showed an upregulation of genes related to B-cell trafficking (CXCL3, CXCR5, CCR6, CCL20) upon treatment. Transient increase of IFNg-induced chemokines (CXCL9, CXCL10) was observed, whereas the pro-inflammatory cytokines (TNFa, IL-2, IL-6, IL-8) were not upregulated. Conclusions: Clinical data and the lack of pro-inflammatory circulating cytokines confirm MP0317 is safe and well-tolerated, while analysis of paired pre- and on-treatment tumor biopsies suggests early evidence of tumor-localized CD40 activation. The current data enables further evaluation in a combination setting. Clinical trial information: NCT05098405 .
Glioblastoma is the most aggressive brain tumor in adults with a median survival below 12 months in population-based studies. The main reason for tumor recurrence and progression is constitutive or ...acquired resistance to the standard of care of surgical resection followed by radiotherapy with concomitant and adjuvant temozolomide (TMZ/RT→TMZ). Here, we investigated the role of microRNA (miRNA) alterations as mediators of alkylator resistance in glioblastoma cells. Using microarray-based miRNA expression profiling of parental and TMZ-resistant cultures of three human glioma cell lines, we identified a set of differentially expressed miRNA candidates. From these, we selected miR-138 for further functional analyses as this miRNA was not only upregulated in TMZ-resistant versus parental cells, but also showed increased expression in vivo in recurrent glioblastoma tissue samples after TMZ/RT→TMZ treatment. Transient transfection of miR-138 mimics in glioma cells with low basal miR-138 expression increased glioma cell proliferation. Moreover, miR-138 overexpression increased TMZ resistance in long-term glioblastoma cell lines and glioma initiating cell cultures. The apoptosis regulator BIM was identified as a direct target of miR-138, and its silencing mediated the induced TMZ resistance phenotype. Altered sensitivity to apoptosis played only a minor role in this resistance mechanism. Instead, we identified the induction of autophagy to be regulated downstream of the miR-138/BIM axis and to promote cell survival following TMZ exposure. Our data thus define miR-138 as a glioblastoma cell survival-promoting miRNA associated with resistance to TMZ therapy in vitro and with tumor progression in vivo.
Abstract Background The coronavirus disease 2019 (COVID-19) pandemic was characterized by rapid evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, affecting viral ...transmissibility, virulence, and response to vaccines/therapeutics. EMPATHY (NCT04828161), a phase 2 study, investigated the safety/efficacy of ensovibep, a multispecific designed ankyrin repeat protein (DARPin) with multivariant in vitro activity, in ambulatory patients with mild to moderate COVID-19. Methods Nonhospitalized, symptomatic patients (N = 407) with COVID-19 were randomized to receive single-dose intravenous ensovibep (75, 225, or 600 mg) or placebo and followed until day 91. The primary endpoint was time-weighted change from baseline in log10 SARS-CoV-2 viral load through day 8. Secondary endpoints included proportion of patients with COVID-19–related hospitalizations, emergency room (ER) visits, and/or all-cause mortality to day 29; time to sustained clinical recovery to day 29; and safety to day 91. Results Ensovibep showed superiority versus placebo in reducing log10 SARS-CoV-2 viral load; treatment differences versus placebo in time-weighted change from baseline were −0.42 (P = .002), −0.33 (P = .014), and −0.59 (P < .001) for 75, 225, and 600 mg, respectively. Ensovibep-treated patients had fewer COVID-19–related hospitalizations, ER visits, and all-cause mortality (relative risk reduction: 78% 95% confidence interval, 16%–95%) and a shorter median time to sustained clinical recovery than placebo. Treatment-emergent adverse events occurred in 44.3% versus 54.0% of patients in the ensovibep and placebo arms; grade 3 events were consistent with COVID-19 morbidity. Two deaths were reported with placebo and none with ensovibep. Conclusions All 3 doses of ensovibep showed antiviral efficacy and clinical benefits versus placebo and an acceptable safety profile in nonhospitalized patients with COVID-19.
Abstract
Background
Ensovibep is a multi-specific DARPin (designed ankyrin repeat protein) antiviral in clinical development for treatment of COVID-19. In the Phase 2 EMPATHY study, ensovibep ...demonstrated greater viral load decline versus placebo. Here we report (1) the efficacy of ensovibep in patients with and without anti-SARS-CoV-2 antibodies at baseline and (2) SARS-CoV-2 mutation emergence data with treatment.
Methods
Eligible ambulatory patients with ≥2 COVID-19 symptoms (onset within 7 days) and positive SARS-CoV-2 rapid antigen test on day of dosing, were randomized (1:1:1:1) to ensovibep (600, 225 or 75 mg) or placebo as single, IV infusion. Chemiluminescent immunoassays were used for antibody detection (SARS-CoV-2 S1/S2 IgG and SARS-CoV-2 IgM). A pre-specified subgroup analysis was performed based on baseline anti-SARS-CoV-2 antibody status. Analysis of changes in viral genome from baseline to post baseline was performed to evaluate treatment-emergent mutations.
Results
Of the patients analyzed, 48.5% had anti-SARS-CoV-2 antibodies at baseline. Baseline log10 SARS-CoV-2 viral load (mean ±SD) was similar across groups ensovibep (all doses) 6.5 ±1.5, placebo 6.2 ±1.5; > 90% were infected with the Delta (B.1.617.2) variant. SARS-CoV-2 viral load reduction up to Day 8 showed similar effects in favor of ensovibep compared with placebo regardless of the presence of anti-SARS-CoV-2 antibodies (Figure 1). Patients in ensovibep 75 mg, 600 mg, and placebo groups had comparable incidences of emergent mutations, with a higher incidence in the 225 mg group. Based on analysis of 70% of the expected viral sequencing data, two mutations in the key binding residues of ensovibep were observed (Y489H and F486L) in a total of three patients treated with ensovibep. These patients either cleared virus by Day 8 or mutations were transient (occurred at a single time point but not later in the course of infection). Figure 1Forest plot of estimated treatment differences and associated 95% confidence intervals in time-weighted change from baseline in log10 SARS-CoV-2 viral load through Day 8 by subgroups for the presence of anti-SARS-CoV-2 antibodies (SARS-CoV-2 S1/S2 IgG and/or SARS-CoV-2 IgM) at baseline.
Conclusion
Ensovibep effectively reduces SARS-CoV-2 viral load regardless of the presence of anti-SARS-CoV-2 antibodies at baseline. Furthermore, there were no emerging mutations of concern, indicating that a single dose administration of ensovibep is associated with minimal selective pressure.
Disclosures
Marc Bonten, MD, PhD, Astra-Zeneca: Advisor/Consultant|Janssen: Advisor/Consultant|Merck: Advisor/Consultant|Novartis: Advisor/Consultant Richa Chandra, MD, Novartis Pharmaceuticals Corporation: Employee Damodaran Solai Elango, MD, Novartis Healthcare Pvt Ltd: Employee Pierre Fustier, PhD, Molecular Partners AG: Employee Kinfemichael Gedif, PhD, Novartis Pharmaceuticals Corporation: Employee Susana Goncalves, MD, Novartis Pharma AG: Employee Awawu Igbinadolor, MD, Novartis: Awawu Igbinadolor reports financial support from different pharmaceutical companies and organizations Jeff Kingsley, DO, MBA, CPI, FACRP, Centricity Research: Other Charles G. Knutson, PhD, Novartis Institutes for BioMedical Research: Employee Petra Kukkaro, PhD, Novartis Pharma AG: Employee Nagalingeswaran Kumarasamy, MD, Novartis: Nagalingeswaran Kumarasamy reports financial support from different pharmaceutical companies and organizations Philippe Legenne, MD, Molecular Partners AG: Employee Martha Mekebeb-Reuter, MD, Novartis: Martha Mekebeb-Reuter reports financial support from different pharmaceutical companies and organizations Krishnan Ramanathan, MD, Novartis Pharma AG: Employee Evgeniya Reshetnyak, PhD, Novartis Pharmaceuticals Corporation: Employee Michael Robinson, PhD, Novartis Institute for Tropical Disease: Employee Jennifer Rosa, MD, Novartis: Jennifer Rosa reports financial support from different pharmaceutical companies and organizations Marianne Soergel, MD, Molecular Partners AG: Employee Vaia Stavropoulou, PhD, Molecular Partners AG: Employee Nina Stojcheva, PhD, Molecular Partners AG: Employee Michael T. Stumpp, PhD, Molecular Partners AG: Employee Andreas Tietz, MD, Novartis Pharma AG: Employee Xiaojun Zhao, PhD, Novartis Institutes for BioMedical Research: Employee Zhaojie Zhang, PhD, 8. Novartis Institutes for BioMedical Research: Employee.