RESUMO
Atovaquone is a substituted 2-hydroxy-naphthoquinone used therapeutically against Plasmodium falciparum (malaria) and Pneumocystis pathogens. It acts by inhibiting the cytochrome bc(1) complex via interactions with the Rieske iron-sulfur protein and cytochrome b in the ubiquinol oxidation pocket. As the targeted pathogens have developed resistance to this drug there is an urgent need for new alternatives. To better understand the determinants of inhibitor binding in the ubiquinol oxidation pocket of the bc(1) complex we synthesized a series of hydroxy-naphthoquinones bearing a methyl group on the benzene ring that is predicted to interact with the nuclear encoded Rieske iron-sulfur protein. We have also attempted to overcome the metabolic instability of a potent cytochrome bc(1) complex inhibitor, a 2-hydroxy-naphthoquinone with a branched side chain, by fluorinating the terminal methyl group. We have tested these new 2-hydroxy-naphthoquinones against yeast and bovine cytochrome bc(1) complexes to model the interaction with pathogen and human enzymes and determine parameters that affect efficacy of binding of these inhibitors. We identified a hydroxy-naphthoquinone with a trifluoromethyl function that has potential for development as an anti-fungal and anti-parasitic therapeutic.
Assuntos
Complexo III da Cadeia de Transporte de Elétrons/química , Naftoquinonas/farmacologia , Animais , Antimaláricos/química , Antimaláricos/uso terapêutico , Atovaquona/química , Atovaquona/uso terapêutico , Bovinos , Citocromos b/química , Citocromos b/efeitos dos fármacos , Citocromos b/genética , Citocromos b/metabolismo , Complexo III da Cadeia de Transporte de Elétrons/metabolismo , Humanos , Cinética , Malária/tratamento farmacológico , Naftoquinonas/química , Plasmodium falciparum/efeitos dos fármacos , Pneumocystis/efeitos dos fármacosRESUMO
There is an urgent need to develop safe, effective, dual-purpose contraceptive agents that combine the prevention of pregnancy with protection against sexually transmitted diseases. Here we report the identification of a group of compounds that on contact with human spermatozoa induce a state of "spermostasis," characterized by the extremely rapid inhibition of sperm movement without compromising cell viability. These spermostatic agents were more active and significantly less toxic than the reagent in current clinical use, nonoxynol 9, giving therapeutic indices (ratio of spermostatic to cytotoxic activity) that were orders of magnitude greater than this traditional spermicide. Although certain compounds could trigger reactive oxygen species generation by spermatozoa, this activity was not correlated with spermostasis. Rather, the latter was associated with alkylation of two major sperm tail proteins that were identified as A Kinase-Anchoring Proteins (AKAP3 and AKAP4) by mass spectrometry. As a consequence of disrupted AKAP function, the abilities of cAMP to drive protein kinase A-dependent activities in the sperm tail, such as the activation of SRC and the consequent stimulation of tyrosine phosphorylation, were suppressed. Furthermore, analysis of microbicidal activity using Chlamydia muridarum revealed powerful inhibitory effects at the same low micromolar doses that suppressed sperm movement. In this case, the microbicidal action was associated with alkylation of Major Outer Membrane Protein (MOMP), a major chlamydial membrane protein. Taken together, these results have identified for the first time a novel set of cellular targets and chemical principles capable of providing simultaneous defense against both fertility and the spread of sexually transmitted disease.
Assuntos
Antibacterianos/farmacologia , Anticoncepcionais/farmacologia , Maleimidas/farmacologia , Quinonas/farmacologia , Espermatozoides/efeitos dos fármacos , Proteínas de Ancoragem à Quinase A/fisiologia , Alquilação , Sobrevivência Celular/efeitos dos fármacos , Chlamydia muridarum/efeitos dos fármacos , Proteínas Quinases Dependentes de AMP Cíclico/fisiologia , Humanos , Masculino , Infecções Sexualmente Transmissíveis/prevenção & controle , Transdução de Sinais , Motilidade dos Espermatozoides/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
Contraception-on-demand refers to contraceptive methods that are only employed when needed, such as barrier or postcoital methods, as opposed to technologies, such as the IUD or pill, where the exposure is continuous irrespective of the risk of pregnancy. The development of women-centered approaches to contraception-on-demand is a high priority in current contraceptive research, with emphasis on the 15- to 25-year-old demographic. Since this cohort of potential users is also at high risk of contracting sexually transmitted disease, topical methods that would provide simultaneous protection against both fertility and infection are of particular interest. This review examines the current strategies that are being pursued to achieve this objective.
Assuntos
Anti-Infecciosos/farmacologia , Anticoncepção/métodos , Infecções Sexualmente Transmissíveis/prevenção & controle , Espermicidas/farmacologia , Adolescente , Adulto , Anti-Infecciosos/uso terapêutico , Feminino , Humanos , Masculino , Espermicidas/uso terapêutico , Adulto JovemRESUMO
There is an urgent clinical need to research novel methods of fertility control that are also protective against sexually transmitted diseases (STDs) such as the human immunodeficiency virus (HIV) or Chlamydia. The most obvious way to generate such a dual-purpose contraceptive method would be to develop safe, effective spermicides that were also active against a wide range of pathogenic organisms. The currently available formulations such as nonoxynol-9, gramicidin and benzalkonium chloride are effective spermicides but are toxic to the vaginal epithelium and do not provide protection against STDs. Over 60 agents are in clinical trials as potentially safer topical spermicides and/or microbicides. Compounds that have reached this stage of development include acid buffers, detergents, dendrimers, non-nucleoside reverse transcriptase inhibitors and anionic polymers. In addition, a number of potential spermicides/microbicides are the subject of preclinical investigation, including beta-cyclodextrin, cyanovirin, porphyrins, cyclotriazadisulfonamides, dermaseptins, short-interfering RNA (siRNA) and HIV antibodies. The chemical principles underlying these disparate approaches and potential avenues for future investigation are discussed.
Assuntos
Anti-Infecciosos/farmacologia , Anticoncepção/métodos , Infecções Sexualmente Transmissíveis/prevenção & controle , Espermicidas/farmacologia , Administração Tópica , Anti-Infecciosos/administração & dosagem , Anti-Infecciosos/uso terapêutico , Feminino , Humanos , Masculino , Espermicidas/administração & dosagem , Espermicidas/uso terapêuticoRESUMO
Atovaquone is a hydroxy-naphthoquinone that is used to treat parasitic and fungal infections including Plasmodium falciparum (malaria), Pneumocystis jivorecii (pneumonia) and Toxoplasma gondii (toxoplasmosis). It blocks mitochondrial oxidation of ubiquinol in these organisms by binding to the ubiquinol oxidation site of the cytochrome bc(1) complex. Failure of atovaquone treatment has been linked to the appearance of mutations in the mitochondrially encoded gene for cytochrome b. In order to determine the optimal parameters required for inhibition of respiration in parasites and pathogenic fungi and overcome drug resistance, we have synthesized and tested the inhibitory activity of novel hydroxy-naphthoquinones against blood stage P. falciparum and liver stage P. berghei and against cytochrome bc(1) complexes isolated from yeast strains bearing mutations in cytochrome b associated with resistance in Plasmodium, Pneumocystis, and Toxoplasma. One of the new inhibitors is highly effective against an atovaquone resistant Plasmodium and illustrates the type of modification to the hydroxy-naphthoquinone ring of atovaquone that might mitigate drug resistance.