title: Drug-membrane interaction studies applied to N'-acetyl-rifabutin.
abstract: This work aims the systematic study of the biophysical interactions of a novel antimycobacterial compound (N'-acetyl-rifabutin, RFB2) with membrane models of different lipid composition and surface charge. Membrane mimetic models were used to evaluate the RFB2's membrane partition, its preferential location across the membrane, and the effect of RFB2 on the biophysical properties of the membrane, which ultimately might be related with the antimycobacterial compound bioavailability and the membrane toxicity. According to the aforementioned, liposomes of dimyristoyl-sn-glycero-phosphocholine (DMPC) and 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) were, respectively, used as mimetic models of human and bacterial cell membranes. The antimycobacterial compound lipophilicity was evaluated by spectroscopic methods, which enabled the determination of the partition coefficient (Kp). To study the RFB2 membrane's location, fluorescence quenching studies and lifetime measurements were executed in liposomes labeled with fluorescent probes. In order to evaluate the changes induced by RFB2 on the membrane biophysical properties, dynamic light scattering (DLS) and steady-state anisotropy were performed. The overall results reveal a strong interaction between RFB2 and the membrane models and allowed the evaluation of its lipophilicity, which is a key molecular descriptor in the characterization of novel potential drugs. Moreover, the higher partition of RFB2 and the more pronounced changes in the biophysical parameters of the negatively charged membrane model suggest that RFB2 has more affinity to the bacterial membrane. For the above-mentioned reasons, this work supports that RFB2 has a potential value as a drug in pharmaceutical formulations used to treat mycobacterial infections.
authors.affiliation: REQUIMTE, Departamento de Ciências Química, Universidade do Porto, Portugal.
authors.forename: Marina
authors.initials: M
authors.lastname: Pinheiro
authors.forename: Mariana
authors.initials: M
authors.lastname: Arêde
authors.forename: João M
authors.initials: JM
authors.lastname: Caio
authors.forename: Cristina
authors.initials: C
authors.lastname: Moiteiro
authors.forename: Marlene
authors.initials: M
authors.lastname: Lúcio
authors.forename: Salette
authors.initials: S
authors.lastname: Reis
mesh_terms: D016880:Anisotropy; D000900:Anti-Bacterial Agents / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008027:Light; D008081:Liposomes; D008567:Membranes, Artificial; D008954:Models, Biological; D010715:Phosphatidylglycerols / Q000378:metabolism; D017828:Rifabutin / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D012542:Scattering, Radiation
chemical_list: D000900:Anti-Bacterial Agents; D005456:Fluorescent Dyes; D008081:Liposomes; D008567:Membranes, Artificial; C000591228:N'-acetylrifabutin; D010715:Phosphatidylglycerols; D017828:Rifabutin; C002773:dimyristoylphosphatidylglycerol; D004134:Dimyristoylphosphatidylcholine
keywords: (2-carboxyethyl)-1,6-diphenyl-1,3,5-hexatriene; 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol; 1,2-dimyristoyl-sn-glycero-phosphocholine; 1,6-diphenyl-1,3,5-hexatriene; DLS; DMPC; DMPG; DPH; DPH-PA; Dynamic light scattering; Fluorescence quenching; HIV; K(SV); K(p); LUVs; Lifetime measurements; MDR-TB; MLVs; MTB; Mycobacterium tuberculosis; N′-acetyl-rifabutin; Partition coefficient; RFB2; Steady-state anisotropy; Stern–Volmer constant; T(m); TB; Tuberculosis; dynamic light scattering; human immunodeficiency virus; large unilamellar vesicles; main phase transition temperature; multidrug-resistant TB; multilamellar vesicles; partition coefficients; tuberculosis