Hydroxy-Propil-β-Cyclodextrin Inclusion Complexes of two Biphenylnicotinamide Derivatives: Formulation and Anti-Proliferative Activity Evaluation in Pancreatic Cancer Cell Models

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive malignancies, with poor outcomes largely due to its unique microenvironment, which is responsible for the low response to drugs and drug-resistance phenomena. This clinical need led us to explore new therapeutic approaches for systemic PDAC treatment by the utilization of two newly synthesized biphenylnicotinamide derivatives, PTA73 and PTA34, with remarkable antitumor activity in an in vitro PDAC model. Given their poor water solubility, inclusion complexes of PTA34 and PTA73 in Hydroxy-Propil-β-Cyclodextrin (HP-β-CD) were prepared in solution and at the solid state. Complexation studies demonstrated that HP-β-CD is able to form stable host–guest inclusion complexes with PTA34 and PTA73, characterized by a 1:1 apparent formation constant of 503.9 M−1 and 369.2 M−1, respectively (also demonstrated by the Job plot), and by an increase in aqueous solubility of about 150 times (from 1.95 µg/mL to 292.5 µg/mL) and 106 times (from 7.16 µg/mL to 762.5 µg/mL), in the presence of 45% w/v of HP-β-CD, respectively. In vitro studies confirmed the high antitumor activity of the complexed PTA34 and PTA73 towards PDAC cells, the strong G2/M phase arrest followed by induction of apoptosis, and thus their eligibility for PDAC therapy.

New Oxaliplatin-Pyrophosphato Analogs with Improved In Vitro Cytotoxicity

“…In order to obtain information on tumor cells’ death, the Annexin V/propidium iodide (PI) assay was used in flow cytometry analyses. Annexin V provides a very sensitive method for detecting cellular apoptosis, while PI is commonly used to detect necrotic or late apoptotic cells (when positive to Annexin too) [ 33 ]. The FITC Annexin V staining was carried out after 24 and 48 h of incubation of HCT116 cells with the Pt-pyrophosphato complexes ( 1 – 4 ) or oxaliplatin/cisplatin (used as reference compounds) at the IC 50 concentrations ( Supplementary Materials, Table S1 ).…”

“…The Annexin V/PI assay was conducted as previously described [ 33 ] in order to investigate the mechanism of cell death induced by the various compounds on HCT116 cells. In particular, after 24 and 48 h of treatment with the compounds at concentrations equal to their IC 50 values calculated by the MTT assay previously described, cells were subjected to Annexin-V FITC/propidium iodide staining, which allowed detection of early and late apoptosis as Annexin V-positive cells, late apoptosis as Annexin V/PI-positive cells, and necrotic cells as PI-positive cells.…”

Microfluidic-Assisted Preparation of Targeted pH-Responsive Polymeric Micelles Improves Gemcitabine Effectiveness in PDAC: In Vitro Insights

“…Human PDAC PANC−1 cells (ATCC-CRL−1469), human pancreas adenocarcinoma ascites metastasis cells AsPC−1 (ATCC-CRL−1682), and undifferentiated human pancreatic carcinoma MIAPaCa-2 cells (ATCC-CRL−1420) were purchased from ATCC (LGC Standard s.r.l., Sesto San Giovanni (MI), Italy), and were cultured as previously reported [ 42 ]. Human pancreatic cancer stllate cancer-associated fibroblasts (CAF08, PC00B5) were purchased from Vitro Biopharma (Golden, Colorado).…”

“…Then, the cells were harvested, washed twice in ice-cold PBS pH 7.4, fixed in 4 mL of 70% ethanol, and stored at −20 °C until analysis. The cell cycle modulation induced by treatments was studied, as previously described [ 42 ], by propidium iodide (PI) staining; the pellet was resuspended in PBS without Ca 2+ and Mg 2+ , containing 1 mg/mL RNase, 0.01% NP40, and 20 µg/mL PI (Sigma); then, FCM analysis was performed by Attune NxT Acoustic Focusing Cytometer (Thermo Fisher Scientific, Waltham, MA, USA). Data were interpreted using the Attune NxT Analysis Software (Thermo Fisher Scientific, Waltham, MA, USA).…”

Cyclodextrin Monomers and Polymers for Drug Activity Enhancement

“…Several biological studies in vitro and in vivo were carried out to express the anticancer activity of complexes containing anticancer compounds [ 17 ]. The anticancer drugs, among plenty of others, such as camptothecin [ 108 ], curcumin [ 109 ], paclitaxel [ 110 ], tamoxifen [ 111 ], resveratrol [ 112 , 113 ], quercetin [ 114 ], temozolomide [ 115 ], doxorubicin [ 116 ], oxaliplatin [ 117 ], β-lapachone [ 118 ], N-biphenylnicotinamides (PTA34 and PTA73) [ 119 ], 13-cis-retinoic acid (13-cis-RA) [ 120 ], oxaliplatin [ 117 ], epothilone A [ 121 ], paclitaxel (PCX) [ 122 ], difluorinated curcumin (CDF) [ 123 ], niclosamide [ 124 ], are complexed with CDs and their derivatives to improve their efficacy, stability, solubility, and bioavailability; reduce their toxicity; and modify their physicochemical peculiarities [ 125 ], in comparison to their uncomplexed forms. After the identification of these new therapeutic anticancer strategies, of particular interest was also the noninclusion complex between CDs and riboflavin (RF) [ 126 ].…”