Solid dispersion with Pluronic F127 was proposed as alternative approach to modify the pharmacologically relevant properties of methotrexate (MTX). Solid dispersion of MTX with Pluronic F127 was prepared by fusion method and characterized by powder X-ray diffraction, thermal analysis, scanning electron microscopy and FTIR spectroscopy with the aim to elucidate the physical state of the dispersed MTX and the nature of the interactions occurring between MTX and the carrier. Effect of Pluronic F127 on solubility, dissolution rate, membrane permeability, and pharmacokinetic parameters was revealed in vitro and in vivo. It was found that physical interactions of MTX with Pluronic F127 are predominant in the solid dispersion. The effect of Pluronic F127 on the MTX solubility and release rate of MTX from the solid dispersion is pH dependent. Apparent solubility of MTX released from the solid dispersion is increased in the acidic medium and remains unchanged in the alkaline medium. In comparison with the pristine MTX, the release of MTX from the solid dispersion is faster in the acidic medium and slower in the alkaline medium. Influence of Pluronic F127 on the membrane permeability of MTX is insignificant. Bioavailability of orally administrated solid dispersion in increased. Results from in vitro and in vivo studies suggested that the pharmacokinetic properties of MTX can be improved by solid dispersion with Pluronic F127. Copyright © 2021 Elsevier B.V. All rights reserved.