Liposomes Containing Drug and Cyclodextrin Prepared by the One-Step Spray-Drying Method

The one-step spray-drying method was applied in the preparation of liposomes containing drug and cyclodextrin (CD). Spray-dried lecithin liposomes, entrapping metronidazole or verapamil alone or together with hydroxypropyl-β-cyclodextrin (HPβCD), were characterized for morphology, size distribution, and drug entrapment efficiency. The main factor influencing the liposomal size was the volume of aqueous medium used for hydration of the spray-dried product. No differences in size or entrapment between liposomes prepared by immediate hydration of dried powder or by hydration after 1 year of powder storage at 4°C were observed. All liposomes were tested for their serum stability. The most stable liposomes (still retaining about 10% of the originally entrapped drug even after 24 hr incubation with serum) were liposomes prepared by the direct spray-drying of the mixture of lipid, drug, and HPβCD.     

Liposome Delivery Systems Containing Ibuprofen

Multilamellar liposome vescicles containing ibuprofen were successfully prepared by hydrating the lipids in the presence of organic solvent. The effects of varying the ratio of lipid to drug; the filter size; and the stirring period during hydration of the dried lipids layer were evaluated. Liposomes sample prepared by using a ratio of 3 lipid: 1 drug gave the highest entrapment efficiency of the drug and released all the drug over 12 hours of testing dissolution. Also the dissolution data showed that the drug release from large liposomes (5.0 μm) was 65.7% after 12 hours; 62.6% from medium size (0.8 pm) and 46.6% from small size liposomes (0.22 μm). Additionally, the increase of the stirring period during hydration of the dried lipid layer with the aqueous phase increased the release of the drug from the liposomes.     

Liposome Delivery Systems Containing Ibuprofen

Abstract

Multilamellar liposome vescicles containing ibuprofen were successfully prepared by hydrating the lipids in the presence of organic solvent. The effects of varying the ratio of lipid to drug; the filter size; and the stirring period during hydration of the dried lipids layer were evaluated. Liposomes sample prepared by using a ratio of 3 lipid: 1 drug gave the highest entrapment efficiency of the drug and released all the drug over 12 hours of testing dissolution. Also the dissolution data showed that the drug release from large liposomes (5.0 μm) was 65.7% after 12 hours; 62.6% from medium size (0.8 pm) and 46.6% from small size liposomes (0.22 μm). Additionally, the increase of the stirring period during hydration of the dried lipid layer with the aqueous phase increased the release of the drug from the liposomes.     

Influence of hydroxypropyl beta-cyclodextrin on the corneal permeation of pilocarpine

The influence of hydroxypropyl β-cyclodextrin (HPβCD) on the corneal permeation of pilocarpine nitrate was investigated by an in vitro permeability study using isolated rabbit cornea. Pupillary-response pattern to pilocarpine nitrate with and without HPβCD was examined in rabbit eye. Corneal permeation of pilocarpine nitrate was found to be four times higher after adding HPβCD into the formulation. The reduction of pupil diameter (miosis) by pilocarpine nitrate was significantly increased as a result of HPβCD addition into the simple aqueous solution of the active substance. The highest miotic response was obtained with the formulation prepared in a vehicle of Carbopol® 940. It is suggested that ocular bioavailability of pilocarpine nitrate could be improved by the addition of HPβCD.     

The Influence of 2-Hydroxypropyl-β-Cyclodextrin on Diffusion Rates and Transdermal Delivery of Hydrocortisone

The influence of 2-hydroxypropyl-β-cyclodextrin (HPβCD) on the permeability of hydrocortisone through semi-permeable cellophane membrane and hairless mouse skin was investigated. When the hydrocortisone concentration was kept constant and the HPβCD concentration in an aqueous vehicle was gradually increased the flux through the semi-permeable membrane was increased until all hydrocortisone had dissolved, after that the flux decreased with increasing HPβCD concentration. A maximum flux was obtained when just enough HPβCD was used to dissolve all the drug in the aqueous vehicle.     

The Influence of Water-soluble Polymers and pH on Hydroxypropyl-β-Cyclodextrin Complexation of Drugs

The effect of water-soluble polymers and ionization of the drug molecules on the cyclodextrin, mainly 2-hydroxypropyl-β-cyclodextrin (HPβCD), solubilization of drugs was investigated. HPβCD has significant solubilizing effect on acetazolamide, prazepam, and sulfamethoxazole in aqueous solutions. All three polymers tested-i.e., hydroxypropyl methylcellulose (HPMC), polyvinylpyrrolidone (PVP), and carboxymethylcellulose-increase the solubilizing effect of HPβCD. The polymers increase the solubilization by increasing the apparent stability constant (K_c of the drug-HPβCD complex. Thus, addition of 0.10% (w/v) HPMC to the aqueous complexation medium results in a 56% increase in K, for the acetazolamide-HPβCD complex and a 200% increase in K_c for the prazepam-HPβCD complex. Addition of 0.25% (w/v) PVP to the complexation medium results in a 138% increase in K for the sulfamethoxazole-HPβCD complex. The HPβCD solubilization of the drugs can also be improved by ionization of the drug molecule through pH adjustments. However, larger improvements of the HPβCD solubilization are obtained when both methods are used simultaneously compared to when either method is used separately.     

Preparation, characterization and in vivo studies of proliposomes containing Cyclosporine A

The present study was undertaken to prepare proliposomes of Cyclosporine A (CsA) to increase its oral bioavailability. The proliposomes were prepared by spraying a solution of CsA, egg lecithin and cremophor EL in methanol-chloroform mixture onto directly compressible lactose (carrier) in a rotary evaporator. A dry free flowing powder of proliposomes was obtained. The dry proliposomal powder was characterized for surface morphology by scanning electron microscopy (SEM). Then the proliposomes were hydrated with distilled water to produce liposomes, which were characterized for particle size distribution, % drug entrapment, and morphological characteristics by transmission electron microscopy (TEM). The liposomes exhibited good entrapment of about 99%. The entrapment of CsA in liposomes was found to be dependent mainly on the drug:lipid ratio. Bioavailability studies were carried out for three different formulations of CsA i.e., free drug suspension; proliposomes derived liposomes and marketed formulation (Pannimun Bioral, Microemulsion) on male SD rats. The results of bioavailability studies indicated that the difference in the mean drug concentration of the free drug and the liposomes was found to be statistically significant (p < 0.05, p value is 0.032). The absorption constant for liposomal product was much greater (10.26 h(-1)) than for free drug solution (1.2 h(-1)) or the marketed sample of microemulsion (2.51 h(-1)) and the volume of distribution was found to be less for liposomes (7629.88 ml/kg) than that of the free drug solution (10971.92 ml/kg) and marketed microemulsion (9012.07 ml/kg). The results of these studies have shown that a stable proliposomal formulation of CsA for oral administration can be prepared which can be easily hydrated into liposomes from which CsA can exert its clinical effects with a better oral bioavailability.     

Preparation,in Vitro and in Vivo Evaluation of Liposomal/Niosomal Gel Delivery Systems for Clotrimazole

Clotrimazole, which is an imidazole derivative antifungal agent, was widely used for the treatment of mycotic infections of the genitourinary tract. To develop alternative formulation for the vaginal administration of clotrimazole to provide sustained and controlled release of appropriate drug for local vaginal therapy, liposomes/niosomes were evaluated as delivery vehicles. To optimize the preparation of liposomes/niosomes with regard to size and entrapment efficiency, multilamellar liposomes/niosomes containing drug were prepared by lipid hydration method. The prepared liposomes/niosomes were incorporated into 2% carbopol gel, and the systems were evaluated for drug stability in phosphate-buffered saline (pH 7.4) and simulated vaginal fluid at 37 ± 1°C. Further, the vesicle gel system was evaluated by antifungal activity and tolerability on tissue level in rat.     

Preparation, in Vitro and in Vivo Evaluation of Liposomal/Niosomal Gel Delivery Systems for Clotrimazole

Clotrimazole, which is an imidazole derivative antifungal agent, was widely used for the treatment of mycotic infections of the genitourinary tract. To develop alternative formulation for the vaginal administration of clotrimazole to provide sustained and controlled release of appropriate drug for local vaginal therapy, liposomes/niosomes were evaluated as delivery vehicles. To optimize the preparation of liposomes/niosomes with regard to size and entrapment efficiency, multilamellar liposomes/niosomes containing drug were prepared by lipid hydration method. The prepared liposomes/niosomes were incorporated into 2% carbopol gel, and the systems were evaluated for drug stability in phosphate-buffered saline (pH 7.4) and simulated vaginal fluid at 37 ± 1°C. Further, the vesicle gel system was evaluated by antifungal activity and tolerability on tissue level in rat.     

Carbamazepine/betaCD/HPMC solid dispersions. II. Physical characterization

Solid dispersions containing carbamazepine (CBZ) associated with β-cyclodextrin (βCD) and/or hydroxypropyl methylcellulose were prepared by two different methods, spray-drying or physical mixture, and characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), infrared (IR) spectroscopy, and x-ray powder diffraction analysis (XRPD) studies. Scanning electron microscopy pictures showed that spray-drying produced a mixture of hollow, spherical, and partially shrunken microparticles of homogeneous materials, whereas the physical mixtures yielded heterogeneous systems in which all individual components could be identified. Thermal and IR analyses suggest the existence of a strong interaction between CBZ and excipients in spray-dried solid dispersions, but no CBZ polymorphic transition was detected by either IR spectroscopy or XRPD analysis after the spray-drying process.     

Preparation, in vitro and in vivo evaluation of liposomal/niosomal gel delivery systems for clotrimazole

Clotrimazole, which is an imidazole derivative antifungal agent, was widely used for the treatment of mycotic infections of the genitourinary tract. To develop alternative formulation for the vaginal administration of clotrimazole to provide sustained and controlled release of appropriate drug for local vaginal therapy, liposomes/niosomes were evaluated as delivery vehicles. To optimize the preparation of liposomes/niosomes with regard to size and entrapment efficiency, multilamellar liposomes/niosomes containing drug were prepared by lipid hydration method. The prepared liposomes/niosomes were incorporated into 2% carbopol gel, and the systems were evaluated for drug stability in phosphate-buffered saline (pH 7.4) and simulated vaginal fluid at 37 +/- 1 degrees C. Further, the vesicle gel system was evaluated by antifungal activity and tolerability on tissue level in rat.     

Overcoming drug-resistant lung cancer by paclitaxel-loaded hyaluronic acid-coated liposomes targeted to mitochondria

Abstract

As a major cause for the inefficiency of cancer chemotherapy, multidrug resistance (MDR) has become a major barrier to cancer treatment. Mitochondrion-orientated transportation of smart liposomes has been developed as a promising strategy to deliver anticancer drugs directly to tumor sites and actively target the mitochondria, so that drugs can interfere with mitochondrial function and facilitate cell apoptosis, overcoming MDR. Herein, we report a novel dual-functional paclitaxel (PTX) liposome system possessing both CD44-targeting and mitochondrial-targeting properties to enhance drug accumulation in mitochondria and trigger apoptosis of drug-resistant cancer cells. Mitochondria-targeting PTX-loaded liposomes were prepared by thin-film hydration and then coated with hyaluronic acid (HA) by electrostatic adsorption. We evaluated the characteristics of the PTX liposomes in vitro, and found that their particle size was about 100?nm and increased to ～140?nm after modification by HA. The entrapment efficiency was larger than 85%, and stability data indicated that the liposomes were physically and chemically stable for at least one week at 4?°C. We further evaluated the intake, mitochondrial targeting, ATP levels, caspase-3 activity measurement, and antitumor actives of the liposomes. The results indicated that HA-coated liposomes with mitochondria targeting had significant inhibitory effects against A549 and A549/Taxol cells, showing them to be a promising means of improving therapeutic efficacy and overcoming MDR in cancer treatment.     

comparative Evaluation of Targeting Efficiency of Charged and Neutral Liposomes of 5-Fluorouracil

Abstract

Liposomes were prepared from egg lecithin by the modified ether injection technique using stearylamine and dicetyl phosphate as the charge-inducing agents. Comparative evaluations of charged and neutral liposomes for drug entrapment, size distribution, in vitro drug release, and in vivo drug targeting were made. The charged liposomes were larger in size and showed better drug entrapment efficiency than the neutral liposomes. However, no difference in the pattern of in vitro drug release was observed. Charged liposomes were found entrapped more in the organs of the reticulo-endothelial system than the neutral liposomes.     

Physicochemical properties of amphotericin B liposomes prepared by reverse-phase evaporation method

The physicochemical properties of phosphatidylcholine-cholesterol liposomes containing amphotericin B and prepared by reverse-phase evaporation method were studied. Uniformly dispersed liposomal suspensions were obtained by employing 3:1 ratio (by volume) of diethyl ether to normal saline, 5 min sonication time at 7°C, and evaporation of diethyl ether at 25°C. Microscopic examination showed that the prepared liposomes were spheroids with unilamellar, oligolamellar, or multilamellar structure. The liposomes containing amphotericin B 2.0 mol% of total lipid led to the highest percentage of drug entrapment. Liposomes with maximum entrapment efficiency were obtained from using 250 µmol of total lipid. The liposomal amphotericin B possessing the highest drug entrapment efficiency (approximately 95%) with particle size range of 1307-1451 nm was the one composed of 1:1 molar ratio of phosphatidylcholine to cholesterol.     

Preparation, optimization, and characterization of topotecan loaded PEGylated liposomes using factorial design

This study reports the development of liposomal system for a potent antitumor drug, topotecan. To achieve this goal conventional and PEGylated liposomes were prepared according to a factorial design by hydration method followed by extrusion. Parameters such as type of lipid, percentage of cholesterol, percentage of phosphatidylglycerols, percentage of polyethylene glycol (PEG)-lipids, and drug to lipid molar ratio were considered as important factors for the optimizing the entrapment and retention of topotecan inside the liposomes. The size and zeta-potential of the PEGylated and conventional liposomes were measured by particle size analyzer and zeta-potentiometer, respectively. The stability and release characteristics of PEGylated liposome loaded topotecan were compared with conventional liposomes and free topotecan. The optimized PEGylated [distearoyl phosphatidylcholine (DSPC)/cholesterol/ distearoyl phosphatidylglycerol (DSPG)/ distearoyl phosphatidylethanolamine-PEG(2000) (DSPE-PEG(2000)); 7:7:3:1.28] and related conventional [DSPC/cholesterol/DSPG; 7:7:3] liposomes showed a narrow size distribution with a polydipersity index of 0.15 and 0.10, an average diameter of 103.0 +/- 13.1 and 95.2 +/- 11.10 nm, and with drug loading of 11.44 and 6.21%, respectively. Zeta-potential was -10 +/- 2.3 and -22 +/- 2.8 mV for PEGylated and conventional liposomes, respectively. The results of stability evaluation showed that the lactone ring of topotecan was notably preserved upon liposome encapsulation. PEGylated liposomes containing topotecan showed a significant decrease (P < 0.001) in release rate in comparison with conventional leptosomes. These results indicate the suitability of PEGylated liposomes in controlling topotecan release. The prepared liposomes (especially PEGylated liposomes) as those described here may be clinically useful to stabilize and deliver topotecan for the treatment of cancer.     

comparative Evaluation of Targeting Efficiency of Charged and Neutral Liposomes of 5-Fluorouracil

Liposomes were prepared from egg lecithin by the modified ether injection technique using stearylamine and dicetyl phosphate as the charge-inducing agents. Comparative evaluations of charged and neutral liposomes for drug entrapment, size distribution, in vitro drug release, and in vivo drug targeting were made. The charged liposomes were larger in size and showed better drug entrapment efficiency than the neutral liposomes. However, no difference in the pattern of in vitro drug release was observed. Charged liposomes were found entrapped more in the organs of the reticulo-endothelial system than the neutral liposomes.     

Effect of water-soluble carriers on dissolution characteristics of nifedipine solid dispersions

Solid dispersions of nifedipine (NP) with polyethylene glycols (PEG4000 and PEG6000), hydroxypropyl-β-cyclodextrin (HPβCD), and poloxamer 407 (PXM 407) in four mixing ratios were prepared by melting, solvent, and kneading methods in order to improve the dissolution of NP. The enhancement of the dissolution rate and the time for 80% NP dissolution T_80% depended on the mixing ratio and the preparation method. The highest dissolution rate and the T_80% as short as 15 min were obtained from PXM 407 solid dispersion prepared by the melting method at the mixing ratio of 1:10. The X-ray diffraction (XRD) patterns of solid dispersions at higher proportions of carriers demonstrated consistent with the results from differential scanning calorimetric (DSC) thermograms that NP existed in the amorphous state. The wettability and solubility were markedly improved in the PXM 407 system. The presence of intermolecular hydrogen bonding between NP and PEGs and between HPβCD and PXM 407 was shown by infrared (IR) spectroscopy.     

Solubility and dissolution rate of progesterone-cyclodextrin-polymer systems

This contribution focused on the solubility improvement of the poorly water-soluble steroid hormone progesterone which, in its natural state, presents a reduced oral bioavailability. In the first part of this study, two simple, reproducible methods that were candidates for use in the preparation of inclusion complexes with cyclodextrins were investigated. Solubility capacities of the progesterone complex with hydroxypropyl-β-CD (HPβ-CD), hydoxypropyl-γ-CD (HPγ-CD), permethyl-β-CD (PMβ-CD), and sulfobutylether-β-CD (SBEβ-CD), prepared by the freeze-drying and precipitation methods, were evaluated by Higuchi phase solubility studies. The results showed that HPβ-CD and PMβ-CD were the most efficient among the four cyclodextrins for the solubilization of progesterone, with the highest apparent stability constants. Therefore, dissolution studies were conducted on these latest progesterone/cyclodextrin complexes and physical mixtures. Two additional natural cyclodextrins, β-CD and γ-CD, were taken as references. Hence, the influence of more highly soluble derivatives of β-CD (HPβ-CD, PMβ-CD) on the progesterone dissolution rate, in comparison to pristine β-CD, alongside an increase in the cavity width for γ-CD versus β-CD, were investigated. The dissolution kinetics of progesterone dissolved from HPβ-CD, PMβ-CD, and γ-CD revealed higher constant rates in comparison to β-CD. Therefore, the aim of the second part of this study was to investigate the possibility of improving the dissolution rate of progesterone/β-CD binary systems upon formation of ternary complexes with the hydrophilic polymer, PEG 6000, as β-CD had the smallest progesterone solubility and dissolution capacity among the four cyclodextrins studied (β-CD, HPβ-CD, HPγ-CD and PMβ-CD). The results indicated that dissolution constant rates were considerably enhanced for the 5% and 10% progesterone/β-CD complexes in PEG 6000.

The interaction of progesterone with the cyclodextrins of interest on the form of the binary physical mixtures, complexes, or ternary complexes were investigated by differential scanning calorimetry (DSC) and Fourier transformed-infrared spectroscopy (FT-IR). The results proved that progesterone was diffused into the cyclodextrin cavity, replacing the water molecules and, in case of ternary systems, that the progesterone β-cyclodextrin was well dispersed into PEG, thus improving progesterone bioavailability for subsequent oral delivery in the same way as derivatized cyclodextrins. The present work proves that ternary complexes are promising systems for drug encapsulation.     

Preparation,Optimization, and Characterization of Topotecan Loaded PEGylated Liposomes Using Factorial Design

This study reports the development of liposomal system for a potent antitumor drug, topotecan. To achieve this goal conventional and PEGylated liposomes were prepared according to a factorial design by hydration method followed by extrusion. Parameters such as type of lipid, percentage of cholesterol, percentage of phosphatidylglycerols, percentage of polyethylene glycol (PEG)-lipids, and drug to lipid molar ratio were considered as important factors for the optimizing the entrapment and retention of topotecan inside the liposomes. The size and zeta-potential of the PEGylated and conventional liposomes were measured by particle size analyzer and zeta-potentiometer, respectively. The stability and release characteristics of PEGylated liposome loaded topotecan were compared with conventional liposomes and free topotecan.

The optimized PEGylated [distearoyl phosphatidylcholine (DSPC)/cholesterol/ distearoyl phosphatidylglycerol (DSPG)/ distearoyl phosphatidylethanolamine-PEG₂₀₀₀ (DSPE-PEG₂₀₀₀); 7:7:3:1.28] and related conventional [DSPC/cholesterol/DSPG; 7:7:3] liposomes showed a narrow size distribution with a polydipersity index of 0.15 and 0.10, an average diameter of 103.0 ± 13.1 and 95.2 ± 11.10 nm, and with drug loading of 11.44 and 6.21%, respectively. Zeta-potential was ?10 ± 2.3 and ?22 ± 2.8 mV for PEGylated and conventional liposomes, respectively. The results of stability evaluation showed that the lactone ring of topotecan was notably preserved upon liposome encapsulation. PEGylated liposomes containing topotecan showed a significant decrease (P < 0.001) in release rate in comparison with conventional leptosomes. These results indicate the suitability of PEGylated liposomes in controlling topotecan release.

The prepared liposomes (especially PEGylated liposomes) as those described here may be clinically useful to stabilize and deliver topotecan for the treatment of cancer.     

Study of the complexation behavior of tenoxicam with cyclodextrins in solution: improved solubility and percutaneous permeability

Complexation of tenoxicam (TEN) with γ-, HPγ-, β-, HPβ-, and Mβ-cyclodextrin (CD) in aqueous solution at pH 7.4 has been investigated using phase solubility diagrams. TEN formed soluble complexes with 1:1 stoichiometry with all the CDs studied, although the inclusion stability constants (K_1:1) obtained had low values. The presence of propylene glycol (PG) in the dissolution medium decreased the stability constants and led to a higher fraction of free drug by competitive displacement and by an increase in the lipophility of the media.

Among the CDs tested, MβCD was chosen for further studies since TEN-MβCD complexes yielded the best results: good solubility and the highest stability constant. The effect of MβCD and PG on the TEN partitioning coefficient was also studied in skin-buffer systems. Although each substance reduced the partitioning value, the combination of PG and MβCD increased this parameter.

The noticeable increase in solubility of the drug found in the presence of MβCD allowed the formulation of carbopol gels with higher doses of TEN and a reduced amount of cosolvent. The presence of MβCD improved the percutaneous penetration of TEN through abdominal rat skin by increasing the solubility of the drug in the vehicle and by affecting the partitioning behavior of TEN in the skin. In addition, TEN retention in the skin was found to be related to the flux values attained with the corresponding gels.