Discovery of a Potent Dual Inhibitor of Aromatase and Aldosterone Synthase.

Estrogen deficiency derived from inhibition of estrogen biosynthesis is a typical condition of postmenopausal women and breast cancer (BCs) patients undergoing antihormone therapy. The ensuing increase in aldosterone levels is considered to be the major cause for cardiovascular diseases (CVDs) affecting these patients. Since estrogen biosynthesis is regulated by aromatase (CYP19A1), and aldosterone biosynthesis is modulated by aldosterone synthase (CYP11B2), a dual inhibitor would allow the treatment of BC while reducing the cardiovascular risks typical of these patients. Moreover, this strategy would help overcome some of the disadvantages often observed in single-target or combination therapies. Following an in-depth analysis of a library of synthesized benzylimidazole derivatives, compound was found to be a potent and selective dual inhibitor of aromatase and aldosterone synthase, with IC values of 2.3 and 29 nM, respectively. Remarkably, the compound showed high selectivity with respect to 11β-hydroxylase (CYP11B1), as well as CYP3A4 and CYP1A2. When tested in cells, showed potent antiproliferative activity against BC cell lines, particularly against the ER+ MCF-7 cells (IC of 0.26 ± 0.03 μM at 72 h), and a remarkable pro-apoptotic effect. In addition, the compound significantly inhibited mTOR phosphorylation at its IC concentration, thereby negatively modulating the PI3K/Akt/mTOR axis, which represents an escape for the dependency from ER signaling in BC cells. The compound was further investigated for cytotoxicity on normal cells and potential cardiotoxicity against ERG and Nav1.5 ion channels, demonstrating a safe biological profile. Overall, these assays demonstrated that the compound is potent and safe, thus constituting an excellent candidate for further evaluation. [Abstract copyright: © 2023 The Authors. Published by American Chemical Society.

New antiproliferative agents derived from tricyclic 3,4-dihydrobenzo[4,5]imidazo[1,2-a][1,3,5]triazine scaffold: synthesis and pharmacological effects

A series of novel 3,4‐dihydrobenzo[4,5]imidazo[1,2‐a][1,3,5]triazine (BIT) derivatives were designed and synthesized. In vitro antiproliferative activity was detected toward two human colorectal adenocarcinoma cell lines (CaCo‐2 and HT‐29) and one human dermal microvascular endothelial cell line (HMVEC‐ d). The most active compounds, namely 2‐4 and 8, were further investigated to clarify the mechanism behind their biological activity. Through immuno fluorescence assay, we identified the target of these molecules to be the microtubule cytoskeleton with subsequent formation of dense microtubule accumulation, particularly at the periphery of the cancer cells, as observed in paclitaxel‐treated cells. Overall, these results highlight BIT derivatives as robust and feasible candidates deserving to be further developed in the search for novel potent antiproliferative microtubule‐targeting agents

Farmaci chemioterapici e antiangiogenici nel trattamento del carcinoma del colon-retto dopo progressione della malattia: studio preclinico farmacodinamico.

Introduzione. Il carcinoma del colon-retto rappresenta il secondo tumore maligno per incidenza e mortalità sia tra gli uomini che tra le donne. Dal 2004, i composti antiangiogenici - in combinazione con farmaci chemioterapici - sono diventati parte del panorama terapeutico del carcinoma del colon-retto metastatico (mCRC). Bevacizumab, un anticorpo monoclonale umanizzato anti-VEGF-A, è stato il primo farmaco antiangiogenico approvato, ma al giorno d’oggi altri inibitori del segnale del VEGF, come regorafenib, aflibercept e ramucirumab, sono stati registrati per la pratica clinica. Tuttavia, sia la resistenza innata che acquisita ai farmaci chemioterapici e antiangiogenici causano nella maggior parte dei pazienti una progressione della malattia durante il trattamento combinato. Alcuni studi clinici hanno dimostrato un beneficio dalla prosecuzione della terapia con bevacizumab dopo la prima progressione, suggerendo che i meccanismi di resistenza al trattamento antiangiogenico potrebbero non essere correlati alla resistenza ai farmaci chemioterapici e che il mantenimento dell'inibizione antiangiogenica con lo switch della chemioterapia a successive linee di trattamento potrebbe rappresentare una valida strategia nel mCRC. Scopo dello studio. Lo scopo del presente studio è la valutazione degli effetti, e dei meccanismi farmacodinamici ad essi correlati, dello switch di farmaci antiangiogenici e chemioterapici, dopo progressione della malattia tumorale, in seguito ad un trattamento di prima linea in un modello preclinico di carcinoma del colon-retto umano. Metodi. Gli studi in vivo sono stati condotti su topi nudi sottoposti a xenotrapianto sottocutaneo di cellule di carcinoma del colon-retto. I trattamenti includevano combinazioni di farmaci con uno switch tra farmaci chemioterapici (irinotecano e 5-fluorouracile) e/o farmaci antiangiogenici (anticorpi anti-VEGF e sunitinib) al momento della progressione tumorale. La valutazione in vitro della citotossicità è stata effettuata su diverse linee cellulari di carcinoma del colon-retto umano esposte a SN-38, metabolita attivo dell’irinotecano, e sunitinib in singolo o in combinazione per 72 ore. L'espressione genica di ABCG2 è stata effettuata in Real-Time PCR e le concentrazioni intracellulari di SN-38 sono state valutate con il sistema HPLC. Risultati. La sostituzione nei trattamenti combinati, al momento della progressione della malattia, del farmaco chemioterapico (da irinotecano a 5-fluorouracile) o del farmaco antiangiogenico (da anticorpi anti-VEGF a sunitinib) ha determinato una nuova risposta, in particolare lo switch alla terapia con irinotecano e sunitinib ha comportato la migliore attività antitumorale. L’analisi immunoistochimica effettuata per i fattori stromali del microambiente tumorale PDGF-C, PlGF, SDF-1α, Tie-2 e VEGFR-2 ha mostrato differenze statisticamente significative tra i tumori al momento della progressione e dopo lo switch della terapia. Inoltre, la combinazione di SN-38 e sunitinib ha prodotto un effetto sinergico sulle linee cellulari di carcinoma del colon-retto, con una significativa inibizione dell'espressione genica di ABCG2 e un aumento delle concentrazioni intracellulari di SN-38. Conclusioni. Le nostre osservazioni precliniche potrebbero avere una rilevanza clinica nei pazienti con tumore del colon-retto, suggerendo lo switch di singoli farmaci chemioterapici o antiangiogenici dopo progressione della malattia per ottenere una nuova risposta tumorale dovuta sia ad una modulazione dei fattori angiogenici del microambiente tumorale, sia ad un effetto diretto sulle cellule tumorali che comporta una possibile variazione delle concentrazioni intracellulari di farmaco

Anti-CD19 monoclonal antibody in combination with metronomic chemotherapy for the treatment of diffuse large B-cell lymphoma: a preclinical study to build a solid rationale for future clinical trials

Introduction. Diffuse large B-cell lymphomas (DLBCL) is the most common lymphoid malignancy in adults, representing almost 30% of all cases of non-Hodgkin’s lymphoma. Metronomic chemotherapy (mCHEMO) can be defined as the frequent, regular administration of drug doses that maintain a low, prolonged, and active, range of plasma concentrations of drugs with a more favorable toxicity profile. mCHEMO in treatment-naïve diffuse large B-cell lymphoma (DLBCL) was recently reported to allow overall and disease-free survivals at 24 months of 54% and 74%, respectively. In non-Hodgkin lymphoma, the combination of targeted therapies with mCHEMO schedules have shown very promising pre-clinical and clinical results. Conversely, the combination of mCHEMO with monoclonal antibodies is almost unexplored in hematological cancers. A novel humanized, anti-CD19 monoclonal antibody (mAb) with an engineered constant fragment (Fc)-domain, has recently been designed to enhance FcγRIIIa binding affinity, for the treatment of B-cell malignancies. Aim of the study. The aim of this study is to evaluate and enhance the activity of an anti-CD19 mAb in concomitant combination with metronomic vinorelbine (mVNR) and etoposide (mETO) on human DLBCL cells and in mouse xenograft models. Methods. In vitro proliferation assay was performed on three human CD19+ DLBCL cell lines, named Toledo, OCI-LY3 and SU-DHL10, exposed to a single dose of anti-CD19 mAb, thrice a week mVNR and daily mETO, alone and their concomitant combination for 144h. Synergism was evaluated by the combination index (CI) and the Loewe additivity model. Apoptosis assay was evaluated by ELISA at 48h. The VNR and ETO intracellular concentration were assessed by UPLC-MS/MS technology. Protein phosphorylation in DLBCL cell lysates was investigated by Luminex analysis. In vivo DLBCL subcutaneous xenograft and systemic tumor models in athymic nude mice were treated with anti-CD19 mAb, mVNR, and their concomitant combination. Results. The single 144h exposure of both mVNR and mETO and anti-CD19 mAb inhibited the DLBCL cell proliferation in a concentration-dependent manner. mETO inhibited the Toledo, OCI-LY3 and SU-DHL10 cell growth with an IC50 of 9.81±1.14 nM, 7.92±3.6 nM and 8.19±0.29 nM, respectively. A higher antiproliferative effect was found after mVNR treatment on Toledo, OCI-LY3 and SU-DHL10 cell lines, as demonstrated by the calculated IC50s of 692.1±134.55 pM, 35.58±12.64 pM and 511.4±133.07 pM, respectively. Interestingly, the anti-CD19 mAb therapy significantly inhibited the Toledo, OCI-LY3 and SU-DHL10 cell proliferation with an IC50 of 1472±351 nM, 906.9±50.87 nM and 57.84±22.14 nM, respectively. In all cell lines, the concomitant treatment anti-CD19 mAb plus mVNR and mETO showed a marked synergism for all the fractions of affected cells (Fa), with a CI1, except for anti-CD19 mAb plus mETO on SU-DHL10 cells. These findings were confirmed by Loewe analysis. A significant pro-apoptotic activity was found in all DLBCL cell-lines treated with anti-CD19 mAb alone for 48h, whereas the combination of anti-CD19 mAb with mVNR and mETO further enhanced apoptosis. Furthermore, the anti-CD19 mAb plus mVNR combination significantly inhibited after 24h the phosphorylation of GSK 3α/β, mTOR, p70S6K, RPS6 and TSC2 proteins in DLBCL cells. Interestingly, anti-CD19 therapy significantly increased the VNR and ETO intracellular concentrations in DLBCL cells after 24h, except ETO levels in SU-DHL10. Finally, in all three subcutaneous DLBCL models, the mVNR and anti-CD19 mAb combination significantly reduced the subcutaneous tumor volumes without causing significant toxicity and significantly increase the overall survival of mice affected by a systemic disease. Conclusions. We report, for the first time, the synergistic activity of anti-CD19 mAb, with mVNR or mETO in DLBCL cells in vitro and in vivo. These results prompt a rapid translation of this combination schedule into future clinical trials

Metronomic Chemotherapy in Pediatric Oncology: From Preclinical Evidence to Clinical Studies

Metronomic chemotherapy (MC) is the frequent, regular administration of drug doses designed to maintain a low, but active, range of concentrations of chemotherapeutic drugs, during prolonged periods of time without inducing excessive toxicities. To date, more than 400,000 children and adolescents under the age of 20 are diagnosed with cancer, per year, with 80% survival in most high-income countries, but less than 30% in low- and middle-income ones. In this review, we summarized the principal preclinical and clinical studies involving the use of MC in the most common pediatric tumors, with an overview of efficacy, toxicity, pharmacokinetic profile, and biomarkers. The best advantages of MC are low toxicity, oral administration and, thus, the feasibility of a more comfortable, home-based treatment, therefore improving the quality of life of the children themselves and of their parents and caregivers. Moreover, MC could represent a valid method to reduce the economic burden of anticancer therapy in the pediatric setting

Metronomic Chemotherapy in Elderly Patients

Purpose of review: This review describes the most relevant studies found in the scientific literature regarding metronomic chemotherapy (MCT) in the geriatric oncology population to support its use as a feasible treatment of care in the frail elderly patients. Recent findings: Recent years have seen a reevaluation of cancer chemotherapeutic drugs and MCT is an emerging schedule in phase II and III clinical trials. Ageing is one of the risk factors for the development of cancer, the incidence of whom increases dramatically in people who live longer. To date, standard oncological protocols involve chemotherapeutic drugs in short cycles of therapy at the maximum tolerated dose (MTD). Although these therapeutic regimens may be successful, they can cause important adverse drug reactions, especially in elderly or frail patients. MCT is a different modality of delivery of chemotherapeutic drugs (frequent low dose for prolonged time) and it looks at the overcoming of the limitations and disadvantages of MTD, in particular the toxicity aspect. We reviewed the experience of clinicians who have used MCT in clinical trials enrolling elderly patients with different cancer types

Complete remission of a diffuse large B-cell lymphoma in a young patient, with severe tuberous sclerosis, treated with metronomic chemotherapy and ibrutinib: a case report

TTuberous sclerosis (TS) is a rare autosomal dominant genetic multisystem disease caused by mutations in either the TSC1 or TSC2 gene and results in the growth of non-cancerous masses in several organs. Diffuse large B-cell lymphoma (DLBCL) is the predominant non-Hodgkin lymphoma in adolescents and young adults. Metronomic chemotherapy (mCHEMO) ca be defined as the frequent, regular administration of drug doses able to maintain a low, but active, range of concentrations of chemotherapeutic drugs, during prolonged periods of time. We present the case of a young woman with severe TS, who developed DLBCL. She was treated consecutively with the mCHEMO schedule R-DEVEC (prednisone, vinorelbine, etoposide, cyclophosphamide plus rituximab) and then ibrutinib, achieving an impressive long-lasting complete remission. In conclusion, alternative treatments could be necessary when comorbidities are present in patients and mCHEMO can be a potential successful therapeutic approach in frail subjects

Active metronomic vinorelbine schedules decrease plasma interleukin-2 levels in mice with Lewis lung carcinoma

The aim of our study was to investigate the effects of metronomic vinorelbine (mVNR) in a tumor model of Lewis Lung (LL) cancer in immunocompetent C57BL/6 mice, looking at the plasma levels of interleukin-2 (IL-2) and interleukin-8 (IL-8). mVNR caused a concentration-dependent antiproliferative effect in vitro on LL/2 cells. The in vivo experiment showed the significant antitumor effects of mVNR at the dose of 4 mg/Kg and 5 mg/Kg, 3 times/week, and the significant dose-dependent decrease of IL-2 concentrations in plasma samples. Conversely, such an effect was not observed for IL-8. A significant decrease in microvessel density was also found at both the active mVNR doses. In conclusion, our study confirmed the activity of mVNR in an immunocompetent model of lung carcinoma and suggest multiple mechanisms of action, including the modulation of IL-2 levels

Synergistic activity of linifanib and irinotecan increases the survival of mice bearing orthotopically implanted human anaplastic thyroid cancer

Anaplastic thyroid cancer (ATC) is the most aggressive form of thyroid cancer, and novel combined therapies are urgently needed to prolong patient survival. No data are currently available on the preclinical activity of the combination of linifanib, a CSF-1R inhibitor, and irinotecan in ATC. The aim of the study was to evaluate the in vitro and in vivo activity of linifanib plus irinotecan. Proliferation and apoptosis assays were performed on 8305C and 8505C human ATC cell lines exposed to SN-38, the active metabolite of irinotecan, linifanib alone, and their concomitant combination. Synergism was evaluated by the combination index method. Quantification of pospho-CSF-1R levels was performed by ELISA. In vivo ATC orthotopic xenografts were treated with the single drugs, or their combination, to evaluate their impact on survival. Histology and immunohistochemistry were performed on ATC tissue samples. Both SN-38 and linifanib inhibited in vitro the proliferation of 8305C and 8505C cells in a concentration-dependent manner, whereas their concomitant treatment revealed a strong synergism in the ATC cells. A significant pro-apoptotic activity was found in both ATC cell lines treated with linifanib alone and in combination with SN-38. Moreover, linifanib significantly decreased the levels of phospho-CSF-1R after 24 h and 72 h in both 8505C and 8305C cells, and this was also observed with the concomitant administration of SN-38. In vivo, the combination of linifanib and irinotecan produced a greater survival result than either monotherapy, and resulted in a significant higher median survival. In some of the mice the combination produced a complete response with a macroscopic disappearance of the disease, as confirmed by histology. In conclusion, the synergistic ATC antitumor activity of linifanib/irinotecan combination significantly increased the survival of ATC affected mice and induced some complete responses, suggesting a potential role of this schedule in ATC patient's treatment