Design, synthesis and characterization of a PEGylated stanozolol for potential therapeutic applications.

Abstract

Stanozolol (STZ) is a drug used to treat serious disorders like aplastic anemia and hereditary angioedema. It is also indicated as an adjunct therapy for the treatment of vascular disorders and growth failures. Encouraging results obtained using animal models demonstrated that STZ increases bone formation and mineralization, thus improving both density and biomechanical properties. Like natural androgens, such as TST and 5α-dihydrotestosterone (5α-DHT), STZ binds androgen receptor (AR) to activate AR-mediated signaling. Despite its therapeutic effects, this synthetic anabolic-androgenic steroid (AAS), or 5α-DHT derivative, due to its high lipophilicity, is poor soluble in water. Thus, to increase the water solubility and stability of STZ, as well as its bioavailability and efficacy, an innovative PEGylated STZ (STZ conjugated with (MeO-PEG-NH2)10kDa, (MeO-PEG-NH)10kDa-STZ) was synthesized. As confirmed by chromatography (RP-HPLC) and spectrometry (ATR-FTIR, 1H NMR, elemental CHNS(O) analysis, MALDI-TOF/TOF) analyses, a very pure, stable and soluble compound was obtained. Acetylcholinesterase (AChE) competitive ELISA demonstrated that the resulting PEGylated STZ competes against biological TST, especially at lower concentrations. Cytotoxicity of increasing concentrations (1, 10, 25 or 50 µM) of STZ and/or (MeO-PEG-NH)10kDa-STZ was also evaluated for up 80 h by performing the MTT assay on human osteosarcoma Saos-2 cells, which express AR and are responsive to STZ. PEGylation mitigated cytotoxicity of STZ, by increasing the cell viability values, especially at higher drug concentrations. Furthermore, these results suggest that (MeO-PEG-NH)10kDa-STZ is a promising and reliable drug to be used in clinical conditions in which TST is required.

• Authors: Cristian Vergallo, Giulia Torrieri, Riccardo Provenzani, Sini Miettinen, Karina Moslova, Markku Varjosalo, Maria Chiara Cristiano, Massimo Fresta, Christian Celia, Hélder A Santos, Felisa Cilurzo, Luisa Di Marzio
• Journal: International journal of pharmaceutics (Int J Pharm) Vol. 573 Pg. 118826 (Jan 05 2020) ISSN: 1873-3476 [Electronic] Netherlands
• PMID: 31715352 (Publication Type: Journal Article)
• Copyright: Copyright © 2019 Elsevier B.V. All rights reserved.

Chemical References

• AR protein, human
• Anabolic Agents
• Androgens
• Receptors, Androgen
• Water
• Polyethylene Glycols
• Testosterone
• Stanozolol

Topics

• Anabolic Agents (chemistry, pharmacokinetics, therapeutic use, toxicity)
• Androgens (chemistry, pharmacokinetics, therapeutic use, toxicity)
• Biological Availability
• Cell Line, Tumor
• Cell Survival (drug effects)
• Chemistry, Pharmaceutical
• Drug Compounding (methods)
• Drug Design
• Drug Stability
• Hormone Replacement Therapy (methods)
• Humans
• Hydrophobic and Hydrophilic Interactions
• Polyethylene Glycols (chemistry)
• Receptors, Androgen (metabolism)
• Solubility
• Stanozolol (chemistry, pharmacokinetics, therapeutic use, toxicity)
• Testosterone (deficiency)
• Toxicity Tests
• Water (chemistry)