RESUMO
BACKGROUND: Novel, in silico-designed anticancer compounds were synthesized in our laboratory namely, 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16). These compounds were designed to have improved bioavailability when compared to their source compound, 2-methoxyestradiol. This theoretically would be due to their increased binding affinity to carbonic anhydrase II, present in erythrocytes. Since the novel compounds under investigation are proposed to be transported within erythrocytes bound to carbonic anhydrase II, the morphological effect which they may exert on whole blood and erythrocytes is of great significance. A secondary outcome included revision of previously reported procedures for the handling of the whole blood sample. The purpose of this study was twofold. Firstly, the ultrastructural morphology of a healthy female's erythrocytes was examined via scanning electron microscopy (SEM) after exposure to the newly in silico-designed compounds. Morphology of erythrocytes following exposure to ESE-15-ol and ESE-16 for 3 minutes and 24 hours at 22°C were described with the use of SEM. The haemolytic activity of the compounds after 24 hours exposure were also determined with the ex vivo haemolysis assay. Secondly, storage conditions of the whole blood sample were investigated by determining morphological changes after a 24 hour storage period at 22°C and 37°C. RESULTS: No significant morphological changes were observed in the erythrocyte morphology after exposure to the novel anticancer compounds. Storage of the whole blood samples at 37°C for 24 hours resulted in visible morphological stress in the erythrocytes. Erythrocytes incubated at 22°C for 24 hours showed no structural deformity or distress. CONCLUSIONS: From this research the optimal temperature for ex vivo exposure of whole blood samples to ESE-15-ol and ESE-16 for 24 hours was determined to be 22°C. Data from this study revealed the potential of these compounds to be applied to ex vivo study techniques, since no damage occurred to erythrocytes ultrastructure under these conditions. As no structural changes were observed in erythrocytes exposed to ESE-15-ol and ESE-16, further ex vivo experiments will be conducted into the potential effects of these compounds on whole blood. Optimal incubation conditions up to 24 hours for whole blood were established as a secondary outcome.
Assuntos
Antineoplásicos/farmacologia , Inibidores da Anidrase Carbônica/farmacologia , Simulação por Computador , Eritrócitos/efeitos dos fármacos , Estradiol/análogos & derivados , Estrenos/farmacologia , Sulfonamidas/farmacologia , Antineoplásicos/farmacocinética , Disponibilidade Biológica , Anidrase Carbônica II/efeitos dos fármacos , Inibidores da Anidrase Carbônica/farmacocinética , Proteínas de Transporte/farmacocinética , Proteínas de Transporte/farmacologia , Descoberta de Drogas , Eritrócitos/ultraestrutura , Estradiol/farmacocinética , Estradiol/farmacologia , Estradiol/toxicidade , Estrenos/farmacocinética , Feminino , Hemólise/efeitos dos fármacos , Humanos , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Pesquisa Qualitativa , Sulfonamidas/farmacocinética , Sulfonamidas/toxicidade , TemperaturaRESUMO
BACKGROUND: Novel, in silico-designed anticancer compounds were synthesized in our laboratory namely, 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16). These compounds were designed to have improved bioavailability when compared to their source compound, 2-methoxyestradiol. This theoretically would be due to their increased binding affinity to carbonic anhydrase II, present in erythrocytes. Since the novel compounds under investigation are proposed to be transported within erythrocytes bound to carbonic anhydrase II, the morphological effect which they may exert on whole blood and erythrocytes is of great significance. A secondary outcome included revision of previously reported procedures for the handling of the whole blood sample. The purpose of this study was twofold. Firstly, the ultrastructural morphology of a healthy female's erythrocytes was examined via scanning electron microscopy (SEM) after exposure to the newly in silico-designed compounds. Morphology of erythrocytes following exposure to ESE-15-ol and ESE-16 for 3 minutes and 24 hours at 22°C were described with the use of SEM. The haemolytic activity of the compounds after 24 hours exposure were also determined with the ex vivo haemolysis assay. Secondly, storage conditions of the whole blood sample were investigated by determining morphological changes after a 24 hour storage period at 22°C and 37°C. RESULTS: No significant morphological changes were observed in the erythrocyte morphology after exposure to the novel anticancer compounds. Storage of the whole blood samples at 37°C for 24 hours resulted in visible morphological stress in the erythrocytes. Erythrocytes incubated at 22°C for 24 hours showed no structural deformity or distress. CONCLUSIONS: From this research the optimal temperature for ex vivo exposure of whole blood samples to ESE-15-ol and ESE-16 for 24 hours was determined to be 22°C. Data from this study revealed the potential of these compounds to be applied to ex vivo study techniques, since no damage occurred to erythrocytes ultrastructure under these conditions. As no structural changes were observed in erythrocytes exposed to ESE-15-ol and ESE-16, further ex vivo experiments will be conducted into the potential effects of these compounds on whole blood. Optimal incubation conditions up to 24 hours for whole blood were established as a secondary outcome.
Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Sulfonamidas/farmacologia , Simulação por Computador , Inibidores da Anidrase Carbônica/farmacologia , Eritrócitos/efeitos dos fármacos , Estradiol/análogos & derivados , Estrenos/farmacologia , Antineoplásicos/farmacologia , Sulfonamidas/toxicidade , Sulfonamidas/farmacocinética , Temperatura , Inibidores da Anidrase Carbônica/farmacocinética , Disponibilidade Biológica , Microscopia Eletrônica de Varredura , Proteínas de Transporte/farmacologia , Proteínas de Transporte/farmacocinética , Anidrase Carbônica II/efeitos dos fármacos , Pesquisa Qualitativa , Eritrócitos/ultraestrutura , Estradiol/toxicidade , Estradiol/farmacologia , Estradiol/farmacocinética , Estrenos/farmacocinética , Descoberta de Drogas , Hemólise/efeitos dos fármacos , Antineoplásicos/farmacocinéticaRESUMO
PURPOSE: To evaluate the performance of 6-O-Lauryl-l-ascorbic acid nanostructures (coagels) as vehicles for acetazolamide (AZM) in ophthalmic administration by in vitro and in vivo experimental tests. METHODS: The systems of coagel + AZM were evaluated in terms of their in vitro release (dialysis membrane), permeability (isolated cornea), pharmacological effectiveness [intraocular pressure (IOP)-reduction in normotensive rabbits], and potential irritant effects. RESULTS: The results concerning AZM permeation were better when vehiculized in coagels compared with ringer solution, which was evident from the AZM steady-state flux and P(app) values (J=1.43 µg/min and P(app)=3.04 cm.s(1)). As a consequence of this increase in permeation, the coagel-AZMs were more effective in lowering the IOP, according to the results obtained from the in vivo assays. Coagels loaded with 0.4% (W/W) of AZM showed a higher hypotensive effect in rabbits compared with the commercial formulation AZOPT(®) (brinzolamide 1%), mainly due to the prolonged effect of the former. In all cases, the intensity of irritation was time dependent. The sodium lauryl sulphate solution (2%) used as a positive control produced serious injury 30 min postadministration. This effect caused irritation, which decreased slowly and even at 180 min, the discomfort was still considerable. However, in the case of coagels, a mild-to-moderate effect was observed. CONCLUSIONS: The incorporation of AZM in coagels seems to improve the ocular bioavailability of this drug. Coagel-AZM 0.4% showed a higher hypotensive effect, with a mild-to-moderate irritant effect. These systems could be administrated in human beings, although more detailed studies still need to be carried out.
Assuntos
Acetazolamida/administração & dosagem , Acetazolamida/farmacocinética , Ácido Ascórbico/análogos & derivados , Inibidores da Anidrase Carbônica/administração & dosagem , Inibidores da Anidrase Carbônica/farmacocinética , Córnea/metabolismo , Portadores de Fármacos/química , Nanoestruturas/química , Acetazolamida/farmacologia , Acetazolamida/toxicidade , Animais , Ácido Ascórbico/química , Disponibilidade Biológica , Inibidores da Anidrase Carbônica/farmacologia , Inibidores da Anidrase Carbônica/toxicidade , Diálise , Sistemas de Liberação de Medicamentos , Pressão Intraocular/efeitos dos fármacos , Soluções Oftálmicas , Permeabilidade , CoelhosRESUMO
IMPORTANCE OF THE FIELD: Acetazolamide (ACZ), a carbonic anhydrase inhibitor (CAI), and other oral CAIs have been an integral part of antiglaucoma therapy for > 40 years. ACZ is used orally for the reduction of intraocular pressure in patients suffering from glaucoma. However, this treatment leads to unpleasant systemic side effects. The answer to the undesirable effects of ACZ is the topical delivery of this drug into the eye, where it could elicit its physiological action. However, the development of a topical formulation of ACZ is limited by its poor ocular bioavailability, which can be largely attributed to its poor penetration coefficient and poor biphasic solubility. AREAS COVERED IN THIS REVIEW: This review offers an overview of different approaches to delivering ACZ to the eye, highlighting the potential of the ternary system ACZ:HP-beta-CD:TEA as a tool for formulating aqueous ACZ eye drop solutions. WHAT THE READER WILL GAIN: A critical analysis is provided to highlight the key issues to design formulations containing hydrophilic cyclodextrins. TAKE HOME MESSAGE: The ACZ:HP-beta-CD:TEA complex is an important new approach to improve the ocular bioavailability of this drug. This approach may be applied to other CAIs in the future.
Assuntos
Acetazolamida/administração & dosagem , Inibidores da Anidrase Carbônica/administração & dosagem , Córnea/metabolismo , Glaucoma/tratamento farmacológico , beta-Ciclodextrinas/química , Acetazolamida/farmacocinética , Administração Tópica , Inibidores da Anidrase Carbônica/farmacocinética , Córnea/efeitos dos fármacos , Desenho de Fármacos , Etanolaminas/química , Feminino , Glaucoma/metabolismo , Humanos , Pressão Intraocular/efeitos dos fármacos , Masculino , beta-Ciclodextrinas/administração & dosagem , beta-Ciclodextrinas/farmacocinéticaRESUMO
In order to enhance the ocular bioavailability of acetazolamide (ACZ), a multicomponent complex with hydroxypropyl-ss-cyclodextrin (HP-ss-CD) and triethanolamine (TEA) was prepared to be applied topically. In vitro corneal permeation across isolated rabbit cornea of proposed ACZ formulations and the marketed AZOPT(R) formulation (1% w/v brinzolamide) was studied. Formulations were also tested for their effect on the intraocular pressure (IOP) in rabbits. (1)H- and (13)C-NMR experiments were undertaken to verify the real inclusion of ACZ in the ACZ-HP-ss-CD-TEA multicomponent complex. The binding of ACZ to HP-ss-CD in the presence of TEA is described. The increase of TEA concentration decreases the apparent equilibrium constant for the ACZ-HP-ss-CD complex. The ternary system ACZ-HP-ss-CD-TEA seemed to be able to reduce IOP in about 30%. This effect was sustained for 4 h after instillation. In vitro corneal permeation studies demonstrated that the ACZ permeation was increased. RMN experiments indicated that TEA can weaken the association between ACZ and HP-ss-CD increasing the drug ocular hypotensive effect by increasing the free drug available for absorption. Our formulations were considered practically non-irritant. These results indicate that the ternary system ACZ-HP-ss-CD-TEA might be a useful tool for formulating aqueous ACZ eye drop solutions.
Assuntos
Acetazolamida/administração & dosagem , Acetazolamida/química , Inibidores da Anidrase Carbônica/administração & dosagem , Inibidores da Anidrase Carbônica/química , Córnea/metabolismo , Etanolaminas/química , beta-Ciclodextrinas/química , 2-Hidroxipropil-beta-Ciclodextrina , Acetazolamida/farmacocinética , Administração Tópica , Animais , Inibidores da Anidrase Carbônica/farmacocinética , Córnea/efeitos dos fármacos , Pressão Intraocular/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Concentração Osmolar , CoelhosRESUMO
Literature data relevant to the decision to allow a waiver of in vivo bioequivalence (BE) testing for the approval of immediate release (IR) solid oral dosage forms containing acetazolamide are reviewed. Acetazolamide's solubility and permeability characteristics according to the Biopharmaceutics Classification System (BCS), as well as its therapeutic use and therapeutic index, its pharmacokinetic properties, data related to the possibility of excipient interactions and reported BE/bioavailability (BA) problems are taken into consideration. The available data on solubility, on oral absorption and permeability are not sufficiently conclusive to classify acetazolamide with certainty. Taking a conservative approach, no biowaiver is considered justified for the registration of new multisource drug products. However, SUPAC level 1 and level 2 postapproval changes and most EU Type I variations can be approved waiving in vivo BE studies.
Assuntos
Acetazolamida/administração & dosagem , Inibidores da Anidrase Carbônica/administração & dosagem , Acetazolamida/farmacocinética , Administração Oral , Inibidores da Anidrase Carbônica/farmacocinética , Formas de Dosagem , Excipientes , Humanos , Solubilidade , Equivalência TerapêuticaRESUMO
Recordemos que el epitelio ciliar consta de dos cepas de células, pigmentadas y no pigmentadas; nuestro concepto es que existe un Sincitium funcional (recordemos que el Sincitium se define como citoplasma con varios núcleos sin límites celulares definidos), esto se refiere a que la membrana basal lateral de las células pigmentadas tiene todos los transportadores que necesita para llevar hacia arriba el cloruro de sodio y luego en consecuencia el agua. Esta membrana tiene el importante intercambio de Na + - K + y obviamente tiene un canal de cloruro y también una etapa de salida de bicarbonato. Nosotros pensamos que esta etapa de salida también puede ser modulada por los inhibidores de la anhidrasa carbónica. Recientemente se mostró en imágenes de video que este concepto es verdadero en el epitelio completo donde ambas cepas de células están juntas; se demostró que los electrolitos son llevados hacia arriba, al interior de la célula, desde las pigmentadas hacia las no pigmentadas y en ellas a la etapa de salida. Por lo tanto hemos visto como el mecanismo del humor acuoso es muy complejo, su secreción comprende apareamiento de transportadores y canales entre células pigmentadas y no pigmentadas y las dos capas de células son funcionalmente una, un Sincitium. Es importante el flujo constante del humor acuoso a través de las cámaras del ojo para una función visual normal. Es necesario un globo ocular formado por una presión intraocular adecuada para mantener la eficacia óptica. Además, el humor acuoso aporta los sustratos necesarios para la función metabólica normal de los tejidos oculares avasculares a los cuales bañan particularmente cristalino, córnea y red trabecular; este flujo de humor acuoso se encarga además de remover los desperdicios metabólicos. Es también un medio para que el iris responda a la luz...