Membrane Transporters Involved in the Antimicrobial Activities of Pyrithione in Escherichia coli

Pyrithione (2-mercaptopyridine-N-oxide) is a metal binding modified pyridine, the antibacterial activity of which was described over 60 years ago. The formulation of zinc-pyrithione is commonly used in the topical treatment of certain dermatological conditions. However, the characterisation of the cellular uptake of pyrithione has not been elucidated, although an unsubstantiated assumption has persisted that pyrithione and/or its metal complexes undergo a passive diffusion through cell membranes. Here, we have profiled specific membrane transporters from an unbiased interrogation of 532 E. coli strains of knockouts of genes encoding membrane proteins from the Keio collection. Two membrane transporters, FepC and MetQ, seemed involved in the uptake of pyrithione and its cognate metal complexes with copper, iron, and zinc. Additionally, the phenotypes displayed by CopA and ZntA knockouts suggested that these two metal effluxers drive the extrusion from the bacterial cell of potentially toxic levels of copper, and perhaps zinc, which hyperaccumulate as a function of pyrithione. The involvement of these distinct membrane transporters contributes to the understanding of the mechanisms of action of pyrithione specifically and highlights, more generally, the important role that membrane transporters play in facilitating the uptake of drugs, including metal–drug compounds.

Targeted Delivery of Zinc Pyrithione to Skin Epithelia

Zinc pyrithione (ZnPT) is an anti-fungal drug delivered as a microparticle to skin epithelia. It is one of the most widely used ingredients worldwide in medicated shampoo for treating dandruff and seborrheic dermatitis (SD), a disorder with symptoms that include skin flaking, erythema and pruritus. SD is a multi-factorial disease driven by microbiol dysbiosis, primarily involving Malassezia yeast. Anti-fungal activity of ZnPT depends on the cutaneous availability of bioactive monomeric molecular species, occurring upon particle dissolution. The success of ZnPT as a topical therapeutic is underscored by the way it balances treatment efficacy with formulation safety. This review demonstrates how ZnPT achieves this balance, by integrating the current understanding of SD pathogenesis with an up-to-date analysis of ZnPT pharmacology, therapeutics and toxicology. ZnPT has anti-fungal activity with an average in vitro minimum inhibitory concentration of 10–15 ppm against the most abundant scalp skin Malassezia species (Malassezia globosa and Malassezia restrica). Efficacy is dependent on the targeted delivery of ZnPT to the skin sites where these yeasts reside, including the scalp surface and hair follicle infundibulum. Imaging and quantitative analysis tools have been fundamental for critically evaluating the therapeutic performance and safety of topical ZnPT formulations. Toxicologic investigations have focused on understanding the risk of local and systemic adverse effects following exposure from percutaneous penetration. Future research is expected to yield further advances in ZnPT formulations for SD and also include re-purposing towards a range of other dermatologic applications, which is likely to have significant clinical impact.

Evaluation of Antimicrobial Effect of Zinc Pyrithione against Airborne Fungi and Bacteria Growth Collected onto New and Loaded HVAC Fibrous Filters

Microbial growth onto HVAC filters was observed in real conditions with possible degradation of the indoor air quality. The filtration performance of marketed antimicrobial filters containing zinc pyrithione was tested under laboratory conditions and compared to that of similar filters with the same classification, F7 (EN779:2002). The filtration performance of the two tested filters during loading with PM10 particles was quantified in an experimental setup with filter pressure drop measurement and particle counting upstream and downstream of the filters. The microbial growth on the new and loaded filters, both contaminated with a microbial airborne consortium composed of two bacteria (Gram-positive and -negative) and fungi, was quantified by colony-forming units after conditioning the filters for a few days under controlled temperature (25 °C) and humidity (50% or 90% relative humidity). The results reveal that there was no degradation of the filtration performance of the filters treated with the antimicrobial agent. The efficiency of the antimicrobial treatment, i.e., the ability to inhibit the growth of microorganisms during the incubation period, was significant with the new filters regarding the fungal growth, but the results demonstrate that the antimicrobial treatment became inefficient with the loaded filters.

Biocides in Antifouling Paint Formulations Currently Registered For Use

Antifouling paints incorporate biocides in their composition seeking to avoid or minimize the settlement and growing of undesirable fouling organisms. Therefore, biocides are released into the aquatic environments also affecting several non-target organisms and, thus, compromising ecosystems. Despite global efforts to investigate the environment occurrence and toxicity of biocides currently used in antifouling paints, the specific active ingredients that have been used in commercial products are poorly known. Thus, the present study assessed the frequencies of occurrence and relative concentrations of biocides in antifouling paint formulations registered for marketing worldwide. The main data were obtained from databases of governmental agencies, business associations and safety data sheets from paint manufacturers around the world. Results pointed out for 25 active ingredients currently used as biocides, where up to six biocides have been simultaneously used in the examined formulations. Cuprous oxide, copper pyrithione, zinc pyrithione, zineb, DCOIT and cuprous thiocyanate were the most frequently ones, with mean relative concentrations of 35.9±12.8 %, 2.9±1.6 %, 4.0±5.3 %, 5.4±2.0 %, 1.9±1.9 % and 18.1±8.0 % (w/w) of respective biocide present in the antifouling paint formulations. Surprisingly, antifouling paints containing TBT as active ingredient are still being registered for commercialization nowadays. These results can be applied as a proxy of biocides that are possibly being used by antifouling systems and, consequently, released into the aquatic environment, which can help to prioritize the active ingredients that should be addressed in future studies.

Zinc pyrithione activates the volume-regulated anion channel through an antioxidant-sensitive mechanism

LRRC8 volume-regulated anion channels (VRACs) play important roles in diverse cell types and may represent therapeutic targets for diseases. To date, however, the pharmacological tools for evaluating the druggability of VRAC have been limited to inhibitors, as no activators of the channel have been reported. We performed a fluorescence-based high-throughput screen (HTS) of 1,184 FDA-approved drugs for compounds that increase VRAC activity. The most potent VRAC potentiator identified was zinc pyrithione (ZPT), which is used commercially for treating dandruff and other skin disorders. In intracellular YFP(F46L/H148Q/I152L)-quenching assays, ZPT potentiates the rate and extent of swelling-induced iodide influx dose-dependently with a half-maximal effective concentration (EC50) of 5.7 µM. Whole-cell voltage-clamp experiments revealed that co-application of hypotonic solution and 30 µM ZPT to HEK293 or HCT116 cells increases the rate of swelling-induced VRAC activation by approximately 10-fold. ZPT potentiates swelling-induced VRAC currents after currents have reached a steady state and activates currents in the absence of cell swelling. Neither ZnCl2 nor free pyrithione activated VRAC, however, treating cells with a mixture of ZnCl2 and pyrithione led to robust channel activation. Finally, the effects of ZPT on VRAC were inhibited by reactive oxygen species (ROS) scavenger NAC and NAD(P)H oxidase inhibitor DPI, suggesting the mechanism of action involves ROS generation. The discovery of ZPT as a potentiator/activator of VRAC demonstrates the utility of HTS for identifying small-molecule modulators of VRAC and adds to a growing repertoire of pharmacological tool compounds for probing the molecular physiology and regulation of this important channel.

Development and Validation of a Complexometric and Potentiometric Titration Method for the Quantitative Determination of Zinc Pyrithione in Shampoo

Most of the pharmaceutical and cosmetic products used for the treatment of dandruff have zinc pyrithione as an active ingredient; therefore; quantifying this component becomes necessary. The purpose of this study was the validation of two simple and fast methodologies in the quantification of zinc pyrithione for shampoo quality control to guarantee consumer safety. The first method comprised a manual complexometric titration, and the second comprised a potentiometric titration performed with an automatic titrator, obtaining sensitivity values of 0.0534% and 0.0038%, respectively, precision expressed in RSD% values below than 1%, and accuracy in recovery percentage greater than 99%. Additionally, both methods were robust when subjected to significant changes in working conditions (temperature and pH) and were selective even in the presence of interferences and degradation products. Finally, the methodologies were adequate to ensure the quality of shampoo to ensure the safety of consumers.