Three-ply walled w/o/w microcapsules formed by a multiple emulsion technique

A new method of microencapsulation is described. Interfacial rheological studies had shown the formation of a rigid bipolymer film at the interface between an aqueous solution of a water-soluble polymer and a non-aqueous solution of an oil-soluble polymer. This led to the idea that small spherical bodies might be formed on making a w/o/w emulsion from these solutions. The present work has shown that ethyl cellulose/acacia microcapsules are formed when the organic solvent ethyl acetate is removed from the multiple emulsion drops. These microcapsules may be obtained as a free-flowing powder.     

Evaluation of process parameters involved in chitosan microsphere preparation by the o/w/o multiple emulsion method

Abstract

Chitosans are interesting biopolymers largely studied for applications in the medical and pharmaceutical fields. In this work, an o/w/o multiple emulsion technique was used for the preparation of hydrophobic drug loaded microspheres. Moreover, the influence of critical variables (concentration of acetic acid in the polymer solution and drug-polymer ratio) on microsphere morphology and drug content was evaluated. Two chitosans of different molecular weights and deacetylation degree were employed; ketoprofen, a non-steroidal anti-inflammatory drug, was chosen as the hydrophobic model drug. The multiple emulsion method produced well-formed microspheres with good yields. Acetic acid concentration in the polymeric solutions influenced particle size and drug content of the microspheres. The highest drug encapsulation efficiencies were obtained for the lowest theoretical drug/chitosan ratio.     

Preparation and characterization of a peptide containing w/o emulsion

Human insulin as a model peptide drug was incorporated into a water-in-oil emulsion prepared by high-pressure homogenization. The formulation of the emulsion was developed with regard to oral compatibility and enhancing properties of the excipients. A fine and stable dispersion of the aqueous phase was achieved by a valid complex of emulsifiers dissolved in a mixture of triglycerides. The resulting droplet size distribution was determined by microscopical means and photon correlation spectroscopy (PCS). The stability of insulin in the formulation was examined by ELISA and HPLC methods over a period of 3 months. Human insulin as a high molecular mass peptide (5800 Da) stayed immunologically active after preparation and the formulation showed high compatibility with the peptide in longer term stability by HPLC. The emulsion was able to protect insulin against gastric degradation in vitro without further encapsulation. Simultaneous incorporation of aprotinin as a suitable protease inhibitor was performed.     

Release of antiseptics from the aqueous compartments of a w/o/w multiple emulsion

A w/o/w multiple emulsion drug carrier system has been developed for local vaginal therapy. To improve its efficacy and to extend the antimicrobial spectrum activity of benzalkonium chloride (CBZ), which is introduced in the external aqueous phase, chlorhexidine digluconate (CHD) was added to the internal aqueous phase of the multiple emulsions. The minimal bactericidal concentrations (MBC) for the association of CHD and CBZ in emulsion were determined towards Escherichia coli and Staphylococcus aureus. The main release mechanism considered for the CHD encapsulated in the inner phase was a swelling-breakdown phenomenon which followed dilution of the emulsion under hypo-osmotic conditions. In order to demonstrate this release, the bactericidal effect of multiple emulsions undiluted and diluted 1-5 and 1-10 in hypo-osmotic conditions at two CHD concentrations was evaluated. To validate and quantify this release, rheological and release kinetics studies were used. The bactericidal activity of combination CBZ-CHD in the emulsion was synergistic on the two bacterial strains and the release of encapsulated CHD in the internal phase was obtained following its dilution in hypo-osmotic conditions. Vaginal administration could be carried out following dilution at 1-5 in sterile water for multiple emulsions containing the lower concentration of CHD.     

Physicochemical characterisation of a drug-containing phospholipid-stabilised o/w emulsion for intravenous administration

Clomethiazole (CMZ) was used as a model drug to be incorporated into an emulsion vehicle. The effects of drug concentration and number of homogenisation steps were evaluated using multiple linear regression. The droplet size, measured as a z-average diameter by photon correlation spectroscopy (PCS), was found to be between 60 and 260 nm in the investigated range of CMZ concentrations, highly dependent on the concentration, but more weakly so on the number of homogenisation steps. Slow-scanning high-sensitivity differential scanning calorimetry (DSC) measurements showed that CMZ depresses the phospholipid chain melting temperature in the emulsion system, whereas (13)C nuclear magnetic resonance (NMR) experiments suggested that the CMZ molecules are to a large extent located in the surface region of the emulsion droplets. This interpretation is compatible with results from NMR self-diffusion measurements, which showed that most of the CMZ molecules are rapidly exchanged between emulsion droplets and the aqueous surrounding. It can be concluded that the surface-active drug CMZ has a significant influence on the characteristics of phospholipid-stabilised emulsions through its ability to interact with the phospholipid interface. Thus, the results underline the importance of characterising drug-lipid interactions for the development of lipid-based formulations.     

Rheological study of w/o/w emulsion by a cone-and-plate viscometer: Negative thixotropy and shear-induced phase inversion

The rheological behaviour of a w/o/w emulsion was examined by a cone-and-plate viscometer. Negative thixotropic flow patterns were observed at lower shear rates. This negative thixotropic behavior was more pronounced and the apparent viscosity increased under increased shear rate, prolonged shearing time, or repeated shear. Further shearing, by raising the shear rate or prolonging the shearing time, rapidly increased the shear stress of the emulsion and induced phase inversion. This phase-inverted emulsion was of the w/o type and in a semi-solid state. The energy dissipated by the flow of the emulsion until the occurrence of phase inversion, the kinetic energy of the emulsion at the phase-inversion point, and the impulse applied to the emulsion by the cone of the viscometer were calculated in order to determine the hydrodynamic parameter determining the phase inversion. The impulse applied by the rotating cone was the parameter determining the phase inversion of the emulsion from a w/o/w to a w/o type characterized by being in the semi-solid state. The negative thixotropic flow and the phase inversion of the w/o/w emulsion to the w/o type induced by shearing were explained as being due to the increase in the volume fraction of the oil droplets by entrapment of water molecules and by coalescence of the oil droplets.     

Effect of formulation and processing variables on the characteristics of microspheres for water-soluble drugs prepared by w/o/o double emulsion solvent diffusion method

80% except for acetaminophen, due to its lower solubility in water and higher solubility in corn oil. The release profile of the drug was pH dependent. In acidic medium, the release rate was much slower, however, the drug was released quickly at pH 7.4. Tacrine showed unexpected release profiles, probably due to ionic interaction with polymer matrix and the shell structure and the highest release rate was obtained at pH 2.0. The prepared microspheres had a sponge-like inner structure with or without central hollow core and the surface was dense with no apparent pores.     

Tabletted polylactide microspheres prepared by a w/o emulsion-spray drying method

An emulsification-spray drying technique is used to prepare poly(D, L-lactic acid) (PDLLA) microparticles loaded with a water soluble, non-steroidal antiinflammatory drug (NSAID), sodium naproxen (NaNPX). The method involves the preparation of a w/o emulsion in which the water soluble drug is dissolved in the aqueous dispersed phase, while the polymer is dissolved in the organic continuous phase. As a comparison, microparticles were prepared by spraying a suspension of the drug into an organic solution of the polymer. The spray-dried particles were characterized using SEM, DSC, XRD and in vitro release tests. The spray-dried product was then compressed (direct compression) to obtain controlled release matrix tablets. All microparticles release NaNPX within 30min. The matrix tablets release the drug in 8-10h; the matrix tablets characterized by the presence of surfactant (due to the emulsion used to obtain the microparticles) have the highest release rate.     

Tabletted polylactide microspheres prepared by a w/o emulsion-spray drying method

An emulsification-spray drying technique is used to prepare poly(D,L-lactic acid) (PDLLA) microparticles loaded with a water soluble, non-steroidal anti-inflammatory drug (NSAID), sodium naproxen (NaNPX). The method involves the preparation of a w/o emulsion in which the water soluble drug is dissolved in the aqueous dispersed phase, while the polymer is dissolved in the organic continuous phase. As a comparison, microparticles were prepared by spraying a suspension of the drug into an organic solution of the polymer. The spray-dried particles were characterized using SEM, DSC, XRD and in vitro release tests. The spray-dried product was then compressed (direct compression) to obtain controlled release matrix tablets. All microparticles release NaNPX within 30 min. The matrix tablets release the drug in 8-10 h; the matrix tablets characterized by the presence of surfactant (due to the emulsion used to obtain the microparticles) have the highest release rate.     

Preparation and evaluation of double-walled microparticles prepared with a modified water-in-oil-in-oil-in-water (w1/o/o/w3) method

Abstract

In this study, a modified water-in-oil-in-oil-in-water (w₁/o/o/w₃) method was developed to prepare double-walled microparticles containing ovalbumin (OVA). The microparticles were characterized with respect to their morphology, particle size, encapsulation efficiency, production yield, thermal properties and in vitro drug release. Microscopy observations clearly showed that microparticles have spherical shape and smooth surface. These microparticles were characterized to have double-walled structure, with a cavity in the centre. By using w₁/o/o/w₃ method, a significant decrease in mean particle size and a significant increase in encapsulation efficiency were obtained. The mean particle size and the encapsulation efficiency of double-walled microparticles were also affected by the changing amount of OVA and mass ratio of polymers. Microparticles prepared with two polymers exhibited a significantly lower initial burst release followed by sustained release compared to microparticles made from poly(d,l-lactide-co-glycolide) 50/50 only. It can be concluded that these microparticles can be a potential delivery system for therapeutic proteins.     

乳化溶剂挥发法制备重组人血管内皮抑制素缓释微球

目的:制备重组人血管内皮抑制素(恩度)缓释微球,并对微球理化性质及体外释放行为进行初步考察。方法:采用乳化溶剂挥发法(W/O/O)制备恩度载药微球;对微球载药量、粒径、突释、体外释放速率及降解行为进行考察,同时利用凝胶电泳初步评价体外释放过程中恩度的完整性。结果:增加聚乳酸-羟基乙酸嵌段共聚物(PLGA)中羟基乙酸的比例、提高PLGA浓度、降低内水相体积、提高理论载药量均增加微球载药能力;降低内水相体积、提高分散速度均减小突释。增加PLGA中羟基乙酸的比例,30 d时累积释放可增加到65%。降解实验说明释放初期微球主要以扩散方式释放恩度,释放后期主要表现为微球的降解。凝胶电泳结果表明微球制备过程对蛋白质聚集性的影响不大。结论:用PLGA作为载体材料制备微球,可以延缓恩度的释放。     

Basic study for stabilization of w/o/w emulsion and its application to transcatheter arterial embolization therapy

Stabilization of w/o/w emulsion and its application to transcatheter arterial embolization (TAE) therapy are reviewed. W/o/w emulsion was stabilized by making inner aqueous phase hypertonic, addition of chitosan in inner phase, and techniques of phase-inversion with porous membrane. Lipiodol w/o/w emulsion for TAE therapy was prepared by using a two-step pumping emulsification procedure. The procedure is so easy that the emulsion could be prepared even during the surgical operation. The deposition after hepatic arterial administration of the emulsion was detected by an X-ray CT scanner. The concentration of epirubicin hydrochloride (EPI) in liver was increased and its residence was prolonged by encapsulating it in the w/o/w emulsion. The toxic effects of EPI and lipiodol on the normal hepatic cells were reduced. The w/o/w emulsion prepared by us is a suitable formulation for the TAE therapy.     

Evaluation of zidovudine encapsulated ethylcellulose microspheres prepared by water-in-oil-in-oil (w/o/o) double emulsion solvent diffusion technique

The preparation of zidovudine-loaded ethylcellulose microspheres by w/o/o double emulsion solvent diffusion method with high entrapment capacity and sustained release is described. A mixed solvent system (MSS) consisting of acetonitrile and dichloromethane in a 1:1 ratio and light liquid paraffin was selected as primary and secondary oil phases, respectively. Span 80 was used as the secondary surfactant for stabilizing the external oil phase. Spherical free flowing microspheres were obtained. The prepared microspheres were characterized by entrapment efficiency, in vitro release behavior, differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). The drug-loaded microspheres showed 32 - 55% entrapment capacity. The in vitro release profile could be altered significantly by changing various processing and formulation parameters to give sustained release of drug from the microspheres. The DSC thermograms confirmed the absence of any drug-polymer interaction. SEM studies showed that the microspheres were spherical and porous in nature. The in vitro release profiles from microspheres of different polymer-drug ratios were best fitted to Higuchi model with high correlation coefficient and the n value obtained from Korsmeyer-Peppas model was ranged between 0.23 - 0.54. The drug release was found to be diffusion controlled mechanism.     

Optimizing drug delivery for enhancing therapeutic efficacy of recombinant human endostatin in cancer treatment

Recombinant human endostatin (rhEndostatin) is uniquely able to target neovascular endothelial cells (ECs) and has the potential for antiangiogenetic and antitumor activities. In this review, we explore experimental approaches and clinical trials focusing on drug delivery of endostatin. Continued endostatin therapy can maintain tumors in a state of dormancy, and no signs of drug-induced resistance have been observed. Prolonged delivery of endostatin may be achieved by using pumps (mini-osmotic pumps) or cell encapsulation systems. The largest benefit from rhEndostatin is expected when drug delivery of rhEndostatin is begun as early as possible. Although endostatin has shown promise in controlling tumor neovasculature, a major problem in pharmacotherapy is the side effects of constant drug administration and the limited half-life of antiangiogenic proteins. Gene therapy offers the advantages of maximizing cost effectiveness and maintaining sustained levels of antiangiogenic factors, which may enhance antitumor efficacy. Therefore, we have investigated recent advances in gene delivery of endostatin for cancer treatment. In recent years, preclinical and clinical data have demonstrated the synergistic effects of rhEndostatin combined with other therapies on inhibiting growth of malignant tumors, with minimal toxicity. rhEndostatin has not been proven to prolong the survival rate of patients challenged with cancer when used as single therapy. Thus, we suggest that the combination of rhEndostatin with chemotherapy, radiotherapy, and biotherapy (i.e., fusion protein, molecular-targeted therapy on cancers, etc.) might provide an optimal strategy for cancer treatment.     

Preparation and evaluation of multiple emulsions water-in-oil-in-water (w/o/w) as delivery system for influenza virus antigens

In this study, investigations have been carried out to prepare adjuvant active delivery systems; multiple water-in-oil-in-water (w/o/w) emulsion formulations, containing influenza virus surface antigen Hemagglutinin (HA). A modified two-stage emulsification method has been used to prepare multiple emulsions. After improving multiple (w/o/w) emulsion formulations; F1: purified antigen solution (PAS)/soybean oil, HCO-40 and span 80/pluronic F-68, F2: PAS and HPbetaCD/soybean oil, HCO-40 and span 80/pluronic F-68, F3: PAS/squalane, HCO-40 and span 80/pluronic F-68, formulations were selected for the stability study that continued for a 3 month duration. To evaluate the stability of these formulations, microscopic observation, osmolarities of the internal and external aqueous phases, pH, globule size and viscosity were determined. SDS-PAGE (silver staining) was used to evaluate HA and the micro-bicinchoninic acid (mBCA) assay was used to determine the in vitro release of antigen from formulations. Immune responses of formulations were investigated in Wistar Albino rats and compared with the immune response raised against the conventional vaccine. These responses were detected with Hemagglutination Inhibition (HAI) assay. The results of this study demonstrated that HA was well entrapped in the multiple (w/o/w) emulsion formulations. Molecular weight and antigenicity of the entrapped HA were not affected by the emulsification procedure. These results suggest that multiple emulsion formulations entrapping influenza antigen may have potential for immunization studies as one of the vaccine delivery system with adjuvant properties.     

In vivo and in vitro characteristics for insulin-loaded PLA microparticles prepared by w/o/w solvent evaporation method with electrolytes in the continuous phase

Insulin-loaded poly(lactide) (PLA) microparticles were successfully prepared by 6% w/v PLA in the organic phase, 10% w/v PVP and varied types of 5%w/v electrolytes in the continuous phase, by using a water-in-oil-in-water emulsion/ solvent extraction technique. Addition of electrolytes such as NaCl, CaCl2 into the external phase significantly improved insulin entrapment efficiency compared to the case of no additives. NaCl was the most effective for obtaining high entrapment efficiency, with microparticle yield 81.2%, trapping efficiencies 49%, insulin-loading level 5.5% w/w and mean particle size 14.8 microm. The distribution (%) of insulin on the PLA microparticles surface, outer layer and core were 8, 37 and 43%, respectively. The cumulative release of insulin had an upper limit of approximately 24% of the insulin load at 24 days. A steady release rate was 0.5 microg insulin/mg microparticles/day of insulin release maintained for 24 days. Total protein-leaking amount was reduced after addition of electrolytes in the continuous aqueous phase. Rabbit glucose levels were evaluated after subcutaneous 20 mg insulin-loaded PLA microparticles or PLA blank microparticles. Study results show that the insulin-loaded PLA microparticles significantly reduced the glucose level than PLA blank microparticles. The insulin-loaded PLA microparticles, physicochemical characterization data and the animal result obtained in this study may be relevant in optimizing the PLA microparticle formulation incorporation and delivery insulin carriers.     

In vivo and in vitro characteristics for insulin-loaded PLA microparticles prepared by w/o/w solvent evaporation method with electrolytes in the continuous phase

Insulin-loaded poly(lactide) (PLA) microparticles were successfully prepared by 6% w/v PLA in the organic phase, 10% w/v PVP and varied types of 5%w/v electrolytes in the continuous phase, by using a water-in-oil-in-water emulsion/solvent extraction technique. Addition of electrolytes such as NaCl, CaCl₂ into the external phase significantly improved insulin entrapment efficiency compared to the case of no additives. NaCl was the most effective for obtaining high entrapment efficiency, with microparticle yield 81.2%, trapping efficiencies 49%, insulin-loading level 5.5% w/w and mean particle size 14.8?µm. The distribution (%) of insulin on the PLA microparticles surface, outer layer and core were 8, 37 and 43%, respectively. The cumulative release of insulin had an upper limit of ～24% of the insulin load at 24 days. A steady release rate was 0.5?µg insulin/mg microparticles/day of insulin release maintained for 24 days. Total protein-leaking amount was reduced after addition of electrolytes in the continuous aqueous phase. Rabbit glucose levels were evaluated after subcutaneous 20?mg insulin-loaded PLA microparticles or PLA blank microparticles. Study results show that the insulin-loaded PLA microparticles significantly reduced the glucose level than PLA blank microparticles. The insulin-loaded PLA microparticles, physicochemical characterization data and the animal result obtained in this study may be relevant in optimizing the PLA microparticle formulation incorporation and delivery insulin carriers.     

Multivesicular emulsion: a novel, controlled-release delivery system for topical dermatological agents

Multivesicular emulsion systems are a new patented technology for topical delivery of pharmaceutical and over-the-counter actives. This novel technology involves the creation of a 2-phase, oil-in-water emulsion system that produces concentric multilamellar spheres of oil and water. Active ingredients can be released from their respective layers upon application to the skin. In addition to a controlled-release of active agents, the multivesicular emulsion base improves the biophysical properties of the skin by reducing transepidermal water loss and enhancing skin hydration. This technology has been applied to 6% salicylic acid formulations that in clinical experience show efficacy with high tolerance in several hyperkeratotic disorders. Subjects' self-assessment was clearly indicative of the excellent cosmetic elegance of the multivesicular emulsion system.