Aqueous Solubility Enhancement for Bioassays of Insoluble Inhibitors and QSPR Analysis: A TNF-α Study

Abstract

The aim of this study is to improve the aqueous solubility of a group of compounds without interfering with their bioassay as well as to create a relevant prediction model. A series of 55 potential small-molecule inhibitors of tumor necrosis factor-alpha (TNF-α; SPD304 and 54 analogues), many of which cannot be bioassayed because of their poor solubility, was used for this purpose. The solubility of many of the compounds was sufficiently improved to allow measurement of their respective dissociation constants (K). Parameters such as dissolution time, initial state of the solute (solid/liquid), co-solvent addition (DMSO and PEG3350), and sample filtration were evaluated. Except for filtration, the remaining parameters affected aqueous solubility, and a solubilization protocol was established according to these. The aqueous solubility of the 55 compounds in 5% DMSO was measured with this protocol, and a predictive quantitative structure property relationship model was developed and fully validated based on these data. This classification model separates the insoluble from the soluble compounds and predicts the solubility of potential small-molecule inhibitors of TNF-α in aqueous solution (containing 5% DMSO as co-solvent) with an accuracy of 81.2%. The domain of applicability of the model indicates the type of compounds for which estimation of aqueous solubility can be confidently predicted.

Structure-Based Discovery of Receptor Activator of Nuclear Factor-κB Ligand (RANKL)-Induced Osteoclastogenesis Inhibitors

“…A key property of the compounds used for screening was solubility, which was assessed as described previously [41]. Herein, we report solubility as observed in 5 mM stock solutions in 100% DMSO, and upon dilutions to 0.3 mM in phosphate-buffered saline (PBS) pH 7.4 containing 5% DMSO.…”

“…Of the 30 compounds obtained, 7 displayed low solubility even at 100% DMSO and were discarded without further evaluation (Low solubility, Table 1). There were 6 compounds that were soluble at stock solutions but displayed limited precipitation at 5% DMSO in PBS [41] and were thus screened at tentative concentrations (Medium solubility, Table 1). Even so, these 6 compounds proved to be inactive in mediating RANKL-induced osteoclastogenesis.…”

Using Physicochemical Measurements to Influence Better Compound Design

“…Methods are emerging, however, to enhance the output of experiments with poorly soluble molecules. 45 The Lipinski Rule of 5…”

Ανακάλυψη αναστολέων για την πρωτεΐνη TNF-α σαν ενώσεις οδηγούς για φάρμακα έναντι της ρευματοειδoύς αρθρίτιδας

“…Ωστόσο, η χρήση τους παρακωλύεται από διάφορους περιορισμούς τόσο από άποψη παραγωγής (υψηλό κόστος) και πρακτικότητας (ενεσοθεραπεία) όσο και από κλινικής άποψης καθώς συχνά εμφανίζουν μη ικανοποιητικό έλεγχο της ασθένειας, ύφεση, ανοσογονικότητα, ανοσοκαταστολή και δυσκολία στη χρήση σε συνδυαστική θεραπεία (Palladino et al, 2003;van Vollenhoven, 2011). Από την άλλη, οι μικρομοριακοί αναστολείς έχουν αναδυθεί σαν μια ελκυστική εναλλακτική για τα βιολογικά φάρμακα εξαιτίας του χαμηλού κόστους και της ευκολίας στην παραγωγή και την ανάπτυξή τους καθώς και λόγω της αυξημένης βιοδιαθεσιμότητάς τους στην από του στόματος χορήγηση (Stanczyk et al, 2008 (Mettou et al, 2017).…”

“…α β Institutional Repository -Library & Information Centre -University of Thessaly 27/09/2021 07:25:07 EEST -44.224.250.200 [88] Η σύνθεση των αναλόγων του SPD304 έγινε από την εταιρία Proactina. Οι ενώσεις αυτές στην πλειοψηφία τους ήταν πάνω από 90% καθαρές (εκτίμηση με HPLC)(Alexiou et al, 2014;Mettou et al, 2017;Papaneophytou et al, 2015). Το SPD304 που χρησιμοποιήθηκε συντέθηκε από την εταιρία Cayman Chemical.…”