Formulation and Optimization of Chitosan-Based Flurbiprofen Nanoparticles for Ocular Delivery: A Design of Experiments Approach.

Introduction:Glimepiride, a BCS class II antidiabetic drug, exhibits poor aqueous solubility leading to limited oral bioavailability. Lipid-based nanocarriers such as solid lipid nanoparticles (SLNs) offer a promising strategy to enhance solubility, stability, and controlled drug release. The present study aimed to develop and statistically optimize glimepiride-loaded SLNs using a Design of Experiments (DOE) approach. Methods:Preformulation studies including FT-IR, UV spectroscopy, and melting point analysis were performed to confirm drug identity and compatibility with excipients. SLNs were prepared using the solvent injection method with Compritol 888 ATO as lipid and Tween-80 as surfactant. A DOE approach (Design Expert software) was employed to optimize formulation variables. The prepared formulations were evaluated for particle size, zeta potential, entrapment efficiency, and in-vitro drug release. Results:All formulations exhibited particle size in the nanorange (59.1–254.1 nm), with zeta potential ranging from −16.89 to 34 mV and entrapment efficiency between 59.2% and 95.3%. The optimized formulation (F6) demonstrated minimal particle size, high entrapment efficiency, and sustained drug release up to 8 hours. DOE analysis confirmed the significant influence of lipid and surfactant concentrations on formulation performance. Conclusion:The optimized SLNs significantly improved the solubility and release profile of glimepiride, indicating their potential as an efficient nanocarrier system for enhancing the therapeutic performance of poorly water-soluble drugs

Tuberculosis (TB) is one of the major health challenge in the world. The current treatment of TB needs daily administration of combined drug therapy for six or more months. Sometime non-adherence and less bioavailability from current therapy develops multidrug resistance, as a result, high dose requirement and subsequent intolerable toxicity are seen. Therefore, nanotechnology gained special attention as it has potential to improve patient compliance, bioavailability and reduction in dosing frequency. The aim of this study is to fabricate alginate-chitosan nanoparticles (AL-CS NPs) under appropriate conditions using ionic gelation method. The use of natural polymers in nanoparticle fabrication has a vast application due to their biodegradability, biocompatibility and nontoxic nature. Ionic gelation method involves the interaction between macromolecules with opposite charged ionizable groups forming polyelectrolyte complex. Hence, it is rational to formulate natural polymerbased sustained release nano-particulate matrix to improve patient adherence, reducing dose frequency and drug toxicity. The formulations were based on 32 factorial designs. Nanoparticles of combined drug (Isoniazid- INH and Pyrazinamide-PYZ) were fabricated using natural polymer. Formulation process involved the use of pregelated sodium alginate followed by ionic gelation with chitosan. Pregelation of sodium alginate included calcium chloride. The effects of sodium alginate concentration and chitosan concentration on particle size, zeta potential, entrapment efficiency and in vitro drug release were studied. Optimized Batch-3s showed particle size 539.7 ± 2.33 nm, zeta potential -26.4 ± 0.55 mV, and entrapment efficiency is 70.21 ± 0.24% and 73.45 ± 0.21% of INH and PYZ, respectively. Dissolution release study of Batch-3s in 7.4 pH phosphate buffer exhibited the initial burst of 5.04 ±0.45% and 19.68 ± 0.87% at 0.25 hrs followed by slow, sustained release of drug 74.53 ± 2.53 and 57.87 ± 2.04% at 10 hrs of INH and PYZ, respectively. It concluded that chitosan (CS) and sodium alginate (AL) concentration are rate-limiting factors in formulation development. Natural polymer based combined drug nano-particulate system could be an innovative and optimistic approach in the treatment of TB.

Objective: To formulate, optimize and evaluate 5-fluorouracil loaded liquorice crude protein nanoparticles for sustained drug delivery using Box-Behnken design.
 Methods: 5-fluorouracil (5-FU) loaded liquorice crude protein (LCP) nanoparticles were prepared by desolvation method using ethanol-water (1:2 ratio), Tween-80 (2%v/v) as stabilizing agent and gluteraldehyde (8% v/v) as cross linking agent. The optimization of prepared nanoparticles was carried out using Box-Behnken design with 3 factors 2 levels and 3 responses. The independent variables were A)5-FU concentration B)LCP concentration and C) sonication time while the responses were R1) Drug entrapment efficiency R2) Drug loading efficiency and R3) Particle size. The correlation between factors and responses were studied through response surface plots and mathematical equations. The nanoparticles were evaluated for FTIR, physicochemical properties like particle size and zeta potential by Photon correlation spectroscopy (PCS) and surface morphology by TEM. The entrapment efficiency, drug loading efficiency and in vitro drug release studies in PBS pH 7.4 (24 h) were carried out. The observed values were found to be in close agreement with the predicted value obtained from the optimization process.
 Results: 5-fluorouracil loaded LCP nanoparticles were prepared by desolvation method, the optimization was carried out by Box-Behnken design and the final formulation was evaluated for particle size (301.1 nm), zeta-potential (-25.8mV), PDI(0.226), with entrapment efficiency (64.07%), drug loading efficiency (28.54%), in vitro drug release (65.2% in 24 h) respectively. The formulated nanoparticles show Higuchi model drug release kinetics with sustained drug delivery for 24 h in pH7.4 buffer.
 Conclusion: The results were proved to be the most valuable for the sustained delivery of 5-Fluorouracil using liquorice crude protein as carrier. 5-FU–LCP nanoparticles were prepared using Tween-80 as stabilizing agent and gluteraldehyde as cross-linking agent to possess ideal sustained drug release characteristics.

The physical properties of nanoparticles may affect the uptake mechanism, biodistribution, stability, and other physicochemical properties of drug delivery systems. This study aimed to first develop a model exploring the factors controlling the nanogel physical properties using a single drug (propranolol), followed by an evaluation of whether these models can be applied more generally to a range of drugs. Size, polydispersity, ζ potential, and encapsulation efficiency were investigated using a design of experiment (DOE) approach to optimize formulations by systematically identifying the effects of, and interactions between, parameters associated with nanogel formulation and drug loading. Three formulation factors were selected, namely, chitosan concentration, the ratio between the chitosan and cross-linker—sodium triphosphate—and the ratio between the chitosan and drug. The results indicate that the DOE approach can be used not only to model but also to predict the size and polydispersity index (PDI). To explore the application of these prediction models with other drugs and to identify the relationship between the drug structure and nanogel properties, nanogels loaded with 12 structurally distinct drugs and 6 structurally similar drugs were fabricated at the optimal condition for propranolol in the model. The measured size, PDI, and ζ potential of the nanogels could not be modeled using distinct DOE parameters for dissimilar drugs, indicating that each drug requires a separate analysis. Nevertheless, for drugs with structural similarities, various linear and nonlinear trends were observed in the size, PDI, and ζ potential of nanogels against selected molecular descriptors, indicating that there are indeed relationships between the drug molecular structure and the performance outcomes, which may be modeled and predicted using the DOE approach. In conclusion, the study demonstrates that DOE models can be applied to model and predict the influence of formulation and drug loading on key performance parameters. While distinct models are required for structurally unrelated drugs, it was possible to establish correlations for the drug series investigated, which were based on polarity, hydrophobicity, and polarizability, thereby elucidating the importance of the interactions between the drug and the nanogels based on the nanogel properties and thus deepening the understanding of the drug-loading mechanisms in nanogels.

This study aimed to develop an improved sustained-release (SR) PLGA microsphere of exenatide using supercritical fluid extraction of emulsions (SFEE). As a translational research, we investigated the effect of various process parameters on the fabrication of exenatide-loaded PLGA microspheres by SFEE (ELPM_SFEE) using the Box-Behnken design (BBD), a design of experiment approach. Further, ELPM obtained under optimized conditions and satisfying all the response criteria were compared with PLGA microspheres prepared using the conventional solvent evaporation (ELPM_SE) method through various solid-state characterizations and in vitro and in vivo evaluations. The four process parameters selected as independent variables were pressure (X 1), temperature (X 2), stirring rate (X 3), and flow ratio (X 4). The effects of these independent variables on five responses, namely the particle size, its distribution (SPAN value), encapsulation efficiency (EE), initial drug burst release (IBR), and residual organic solvent, were evaluated using BBD. Based on the experimental results, a desirable range of combinations of various variables in the SFEE process was determined by graphical optimization. Solid-state characterization and in vitro evaluation revealed that ELPM_SFEE improved properties, including a smaller particle size and SPAN value, higher EE, lower IBR, and lower residual solvent. Furthermore, the pharmacokinetic and pharmacodynamic study results indicated better in vivo efficacy with desirable SR properties, including a reduction in blood glucose levels, weight gain, and food intake, for ELPM_SFEE than those generated using SE. Therefore, the potential drawback of conventional technologies such as the SE for the preparation of injectable SR PLGA microspheres could be improved by optimizing the SFEE process.

Encapsulation of urea was performed in chitosan microspheres via emulsification followed by cross-linking with genipin, a natural cross-linker. The microspheres were prepared by varying different parameters, e.g., concentrations of chitosan, urea and cross-linker. The effect of these parameters on urea loading (%), urea content (%), entrapment efficiency (%) and release rate was studied. Higher amount of chitosan (1.0 g) and cross-linker concentration (0.75 mmol/g of chitosan) produced entrapment efficiencies of 99.0 and 78.5 %, respectively. Release rate was found to be dependent on the concentrations of urea, chitosan, cross-linker and temperature of the release medium. Higher concentration of loaded urea enhanced the release rate, whereas higher concentrations of chitosan and cross-linker reduced it. Higher temperature of the release medium improved the release rate. It was found that water uptake (%) increased through the increase of concentrations of urea and chitosan and decrease of that of cross-linker. Fourier transform infrared (FTIR) spectroscopy indicated the incorporation of urea in the chitosan microspheres. There was no significant interaction between chitosan and urea as evidenced by FTIR study. Surface of the urea-loaded microspheres appeared coarser and rough compared to that of unloaded microspheres as revealed by scanning electron microscopy.

Breast cancer is the most common cancer in females and ranked second after skin cancer. The use of natural compounds is a good alternative for the treatment of breast cancer with less toxicity than synthetic drugs. The aim of the present study is to develop and characterize hybrid Apigenin (AN) Nanoparticles (NPs) for oral delivery (AN-NPs). The hybrid AN-NPs were prepared by the self-assembly method using lecithin, chitosan and TPGS. Further, the NPs were optimized by Box-Behnken design (3-factor, 3-level). The hybrid NPs were evaluated for particle size (PS), entrapment efficiency (EE), zeta potential (ZP), and drug release. The optimized hybrid NPs (ON2), were further evaluated for solid state characterization, permeation, antioxidant, cytotoxicity and antimicrobial study. The formulation (ON2) exhibited small PS of 192.6 ± 4.2 nm, high EE 69.35 ± 1.1%, zeta potential of +36.54 mV, and sustained drug release (61.5 ± 2.5% in 24 h), as well as significantly (p < 0.05) enhanced drug permeation and antioxidant activity. The IC50 of pure AN was found to be significantly (p < 0.05) lower than the formulation (ON2). It also showed significantly greater (p < 0.05) antibacterial activity than pure AN against Bacillus subtilis and Salmonella typhimurium. From these findings, it revealed that a hybrid AN polymeric nanoparticle is a good carrier for the treatment of breast cancer.

Sparfloxacin-loaded PLGA nanoparticles for sustained ocular drug delivery

The present study aimed to develop and optimize chitosan coated solid lipid nanoparticles (chitosan-SLNs) encapsulated with methazolamide. Chitosan-SLNs were successfully prepared by a modified oil-in-water emulsification-solvent evaporation method with glyceryl monostearate as the solid lipid and phospholipid as the surfactant. Systematic screening of formulation factors was carried out. The optimized formula for preparation was screened by orthogonal design as well as Box-Behnken design with entrapment efficiency, particle size and zeta potential as the indexes. The entrapment efficiency of the optimized formulation (methazolamide-chitosan-SLNs) prepared was (58.5±4.5)%, particle size (247.7±17.3) nm and zeta potential (33.5±3.9) mV. Transmission electron microscopy showed homogeneous spherical particles in the nanometer range. A prolonged methazolamide in vitro release profile was obtained in the optimized chitosan-SLNs suspension compared with methazolamide solution. No ocular damages were observed in the susceptibility test on albino rabbits. The results suggest that the combination of orthogonal design and Box-Behnken design is efficient and reliable in the optimization of nanocarriers, and chitosan-SLNs is a potential carrier for ophthalmic administration.

Box-Behnken Designed Fluconazole Loaded Chitosan Nanoparticles for Ocular Delivery The present study aimed to develop chitosan nanoparticles containing Fluconazole for ocular delivery using Box-Behnken design. The Fluconazole loaded chitosan nanoparticles were prepared by an ionic gelation method using Sodium tripolyphosphate (NaTPP) as cross linking agent. The effect of the factors - concentration of chitosan (x1), concentration of NaTPP (x2) and volume of NaTPP (x3) was studied on release of drug from nanoparticles. The results revealed that entrapment efficiency was highest at low level of chitosan concentration, high level of NaTPP concentration and low levels of NaTPP volume. The optimized batch (NP 3) showed encapsulation efficiency of 63.1%, particle size of 471 nm, ovoid shape surface morphology and in vitro cumulative percentage of drug release 39.19% in 7 h.

Formulation and statistical optimization of gastric floating alginate/oil/chitosan capsules loading procyanidins: in vitro and in vivo evaluations

Ciprofloxacin is a commonly used antibiotic for treatment of bacterial conjunctivitis. The conventional eye drop dosage form is the widely used mode of treatment, but it has low corneal residence time. This drawback can be overcome by developing a bioadhesive noisome system (chitosan-coated) for enhanced corneal residence time. The niosomes were prepared by thin-film hydration technique and optimized by using Box-Behnken statistical design software. Cholesterol (A), Span 60 (B), and sonication time (C) were selected as independent variables, whereas vesicle size (Y1 in nm), entrapment efficiency (Y2 in %), and drug release (Y3 in %) were chosen as dependent variables. The vesicle size, entrapment efficiency, and drug release of optimized CIP niosomes (CIP-NSMopt) were found to be 180.34 ± 5.13nm, 78.32 ± 4.49%, and 82.87 ± 4.01% (in 12h), respectively. Further CIP-NSMopt was coated with different chitosan concentrations (0.1 to 0.3%) to enhance mucoadhesion. Finally, optimized chitosan-coated niosomes (chitosomes; CIP-CHTopt) showed a vesicle size of 210.65 ± 2.76nm, zeta potential of - 35.17 ± 2.25Mv, and PDI of 0.221. CIP-CHTopt exhibited sustained release profile (75.31% in 12h) with the Korsmeyer-Peppas kinetic model (R2 = 0.980). The permeation study showed 1.79-fold enhancements in corneal permeation compared with marketed CIP eye drop. The hen's egg chorioallantoic membrane (HET-CAM) study showed 0 scores (no irritation), and it was further confirmed by corneal hydration and histopathology study. The antimicrobial study exhibited a significant high zone (P < 0.05) of inhibition against tested organism. Our findings demonstrated that chitosan-coated niosomes are a promising drug carrier to enhance corneal contact time and treatment of bacterial conjunctivitis.

Bortezomib (BTZ) is a proteasome inhibitor approved for the treatment of multiple myeloma. It is under investigation for use in the treatment of other solid tumors, such as breast and prostate cancers. BTZ is known for its high potency, limited aqueous solubility, and systemic toxicity. These characteristics restrict its clinical application. This study aims to formulate BTZ in the form of bovine serum albumin (BSA) nanoparticles (NPs) to enhance its anticancer targeting. The Box-Behnken experimental design was employed to optimize the formulation of BTZloaded BSA NPs. BTZ BSA NPs were prepared through the desolvation method by using glutaraldehyde (Glut) as a crosslinker. The Box-Behnken design was employed as an experimental design tool to investigate the impact of formulation parameters, specifically the amount of BTZ, BSA, and antisolvent volume, on outcomes, including particle size (PS), entrapment efficiency (EE%), and polydispersity index (PDI). The morphology of the prepared NPs was examined using a transmission electron microscope (TEM), while a compatibility study was conducted using Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The optimized formulation resulted in a mean PS, EE%, and PDI of 74 nm, 68%, and 0.007, respectively. The BTZ BSA NPs exhibited a sustained in vitro release profile over 24 hours. TEM revealed that the prepared NPs had an almost spherical morphology. FTIR and DSC revealed that BTZ was encapsulated well within the prepared NPs with minimal decomposition and/or degradation. The optimized BTZ BSA NPs exhibited uniform nanosize, high entrapment efficiency, and sustained drug release, confirming the effectiveness of Box-Behnken optimization. Encapsulation within the cross-linked albumin matrix protected BTZ from hydrolytic degradation, enhanced its stability, and provided controlled release, supporting its potential for improved anticancer activity and reduced systemic toxicity. The use of experimental design is a valuable tool for optimizing the formulation of BTZ BSA NPs, achieving the required PS, high EE%, and low PDI.

Introduction: This study aims to develop and optimize a phytosome-based drug delivery system encapsulating the hydroethanolic extract of Ipomoea cairica (IPc-ex) to substantiate its traditional applications in treating inflammation. Material and Methods: The crude plant material was subjected to extraction using various solvents including hydroethanolic, chloroform, ethyl acetate and n-hexane. Among these, hydroethanolic solvent yielded the highest extraction efficiency. IPc-ex-PS were formulated using the thin film hydration method. The Box-Behnken design was utilized to optimize the formulation and process parameters. Independent variables were taken as Phospholipid concentration (A), temperature (B) and reaction time (C) while dependent variables were Particle size and % Entrapment efficiency. Characterization of the phytosomes was conducted using zetasizer, FT-IR and TEM analyses. The in-vitro drug release from IPc-ex phytosomes was assessed using dialysis bag technique. The anti-inflammatory effect of hydroethanolic extract (IPc-ex) and hydroethanolic extract loaded phytosomes (IPc-ex-PSopt) was evaluated through the protein denaturation study. Results: The phenolic content was found to be 43.15±1.28 mg Gallic acid equivalent/g and flavonoid content was 26.39±1.56 mg Quercetin equivalent/g in hydroethanolic extract of Ipomoea cairica. The optimized batch of hydroethanolic extract loaded phytosomes (IPc-ex-PSopt) exhibited the smallest average size of 118 nm. The entrapment efficiency and zeta potential of 79.59±0.21% and -25.8mV, respectively. The percentage drug release of the active constituent from IPc-ex-PSopt was 87.75±1.27% at the end of 12 hr. The optimized formulation (IPc-ex-PSopt) exhibited a concentration-dependent inhibitory effect with a maximum inhibition of 87.49±0.79% at 200 μg/ml and 37.61±0.75% at 50 μg/ml significantly when compared with standard drug, Diclofenac sodium. Discussion: The findings demonstrate that Ipomoea cairica extract-loaded phytosomes (IPc-ex-PSopt) can serve as an effective delivery system with high encapsulation efficiency and controlled release. Conclusion: The study suggests that IPc-ex-PSopt exhibit potential efficacy in mitigating inflammatory conditions, as evidenced by these experimental results.

BackgroundThe overall objective was to prepare a highly accurate nanocarrier system of mesalamine for the treatment of ulcerative colitis with increased therapeutic efficacy and targeting. In the formulation of nanocarrier systems, optimization is a critical process for understanding nanoformulation variables and quality aspects. The goal of the present work was to determine the effect of independent variables, i.e., the concentrations of chitosan, carboxymethyl inulin (CMI), and the drug on the response variables, i.e., particle size and percent entrapment efficiency of the mesalamine-loaded nanoparticle using the Box Behnken design (BBD). The correlation between the independent and dependent variables was investigated using the Design Expert generated mathematical equations, contour, and response surface designs.ResultAn optimized batch was developed using the ionotropic gel method with selected independent variables (A: + 1 level, B: 0 level, C: − 1 level) and the developed nanoparticles had a particle size of 184.18 nm, zeta potential 26.54 mV, and entrapment efficiency 88.58%. The observed responses were remarkably similar to the predicted values. The morphological studies revealed that the formulated nanoparticles were spherical, and the results of the FTIR and DSC studies indicated the drug-polymer compatibility. The nanoparticle showed less than 5% release in the pH 1.2. In the colonic region (pH 7.4), more than 80 % of the medication was released after 24 h. The kinetics study showed that the Higuchi and Korsemeyer-Peppas models had R2 values of 0.9426 and 0.9784 respectively, for the developed formulation indicating linearity, as revealed by the plots. This result justified the sustained release behavior of the formulation.ConclusionThe mesalamine-loaded chitosan-CMI nanoparticle has been successfully developed using the ionotropic gelation method. The nanoparticles developed in this study were proposed to deliver the drug to its desired site. The developed nanoparticles were likely to have a small particle size with positive zeta potential and high percent drug entrapment. It could be stated from the results that BBD can be an active way for optimizing the formulation and that nanoparticles can be a potential carrier for delivering therapeutics to the colon.