Carrier For Oral Delivery Of Insulin Research Articles

Efficacy of oral insulin nanoparticles for the management of hyperglycemia in a rat model of diabetes induced with streptozotocin.

Insulin is the cornerstone of treatment in type 1 diabetes mellitus. However, because of its protein structure, insulin has to be administered via injection, and many attempts have been made to create oral formulations, especially using nanoparticles (NPs). The aim of this study was to compare the hypoglycemic effect of insulin-loaded NPs to that of subcutaneous insulin in an in vivo rat model of diabetes. We used biodegradable D-α-tocopherol polyethylene glycol succinate-emulsified, chitosan-capped poly(lactic-co-glycolic acid) NPs loaded with soluble human insulin in a dose of 20 IU/kg body weight, and examined the physical characteristics of NPs in vivo and in vitro. Serum glucose levels were reduced after 6 h, but the difference was not significant compared to subcutaneous insulin; at 12 h and 24 h, insulin levels were significantly higher in rats treated with NPs than in rats treated with subcutaneous insulin. There was no significant difference in serum insulin levels at 12 h and 24 h compared to non-diabetic rats. Our findings suggest that chitosan-based NPs are able to maintain good glycemic control for up to 24 h and can be considered a potential carrier for oral insulin delivery.

Hypoglycemic Effect of Orally Delivered Insulin Nanoparticles Compared to Subcutaneous Recombinant Human Soluble Insulin in Hyperglycemic Male Rats

Background: Insulin is available as an injectable drug and remains the mainstay of therapy. Researchers have attempted to develop an oral formulation of insulin, particularly utilizing nanoparticles (NPs). Objective: To assess the efficacy and safety of insulin-loaded D-α-Tocopherol polyethylene glycol succinate (TPGS)-emulsified PEG-capped PLGA NP in comparison to insulin-loaded PLGA NP and subcutaneous (SC) insulin in an in vitro and in vivo diabetic rat model. Methods: Two biocompatible and biodegradable NPs were used, in which 20 IU/kg of recombinant human soluble insulin was incorporated. NP1 was PLGA-loaded with human insulin, while NP2 was PLGA-PEG-TPGS-loaded with human insulin. The physical characteristics of the NPs were examined in an in vitro and in vivo study on a hyperglycemic rat model for a 24-hour duration. Results: For the first 3 hours, SC insulin showed a better reduction in serum glucose levels (SG) compared to both NP1 and NP2. A smooth, sustained reduction in SG was observed and maintained till the end of 24 hours with both NPs. NP1 maintained SG reduction for 6 hours before experiencing an increase, while NP2 demonstrated superior sustained reduction in SG beyond the 12-hour evaluation period. Conclusions: PLGA-PEG-TPGS NP can act as a potential carrier for oral insulin delivery and maintain good glycemic control for up to 24 hours.

Construction and functional evaluation of oral long-acting insulin hydrogel microparticles based on physical and chemical double crosslinking

The objective of this study was to enhance the convenience and effectiveness of diabetes treatment by developing hydrogel microparticles as an oral insulin delivery system, aiming to reduce the necessity for frequent treatments. The hydrogel microparticles were prepared with polysaccharides through a combination of physical and chemical crosslinking method, they achieved good results in insulin loading efficiency (70 %), insulin release efficiency (98 %) and sustained release time (>20 h). The effective transmembrane transport was validated using an intestinal epithelial cell model, which demonstrated a continuous hypoglycemic effect lasting from 6 to 26 h in a type 2 diabetes mouse model. Additionally, the relative bioavailability of insulin reached 30.14 ± 2.62 %, representing a significant breakthrough in the field of oral insulin delivery carriers. Furthermore, oral insulin hydrogel exhibited a substantial improvement in insulin resistance, organ damage, and diabetes-related complications stemming from hyperglycemia. These compelling findings underscore the potential of hydrogel microparticles as a cost-effective and valuable strategy for oral drug delivery in diabetes treatment.

Multi-functional self-assembly nanoparticles originating from small molecule natural product for oral insulin delivery through modulating tight junctions

BackgroundOral administration of insulin (INS) could be absorbed into systemic circulation only if the carrier protected it from the hostile gastrointestinal conditions. However, traditional macromolecular carriers have not totally overcome challenges in addressing these biological barriers.ResultIn this study, inspired by small molecule natural products (SMNPs), we demonstrate the multi-functional self-assembly nanoparticles (BA-Al NPs) originating from baicalin (BA) and AlCl3 through coordination bonds and hydrogen bonds. As a novel carrier for oral insulin delivery (INS@BA-Al NPs), it displayed effective capacity in pH stimuli-responsive insulin release, intestinal mucoadhesion and transepithelial absorption enhance. Meanwhile, BA improved the paracellular permeability for insulin absorption, because of its downregulation at both mRNA and protein level on internal tight junction proteins. In vivo experiments exhibited remarkable bioavailability of INS and an ideal glucose homeostasis in the type I diabetic rat model.ConclusionThis study offers a novel frontier of multi-functional carriers based on SMNPs with self-assembly character and bioactivity, which could be a promising strategy for diabetes therapy.Graphical

Prediction of the enhanced insulin absorption across a triple co-cultured intestinal model using mucus penetrating PLGA nanoparticles.

Insulin is a protein macromolecule used to treat diabetes mellitus. Currently, insulin requires multiple daily subcutaneous (SC) injections to control blood sugar in diabetics. Thus, reducing the patients' compliance and adherence to medication as SC route is invasive. Insulin is poorly absorbed through intestinal epithelium because it is a large and hydrophilic molecule, degraded by proteases, and due to the presence of mucosal biophysical barrier. Herein, insulin was encapsulated into different poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) formulations prepared by microfluidic technique, which were further appended with heparin sulfate for oral insulin delivery. The average particle size was ca. 200nm, PDI at ca. 0.3 and the zeta potential at ca. -20mV. The maximal achieved association efficiency of insulin was~55% at 3% theoretical loading. PLGA exhibited a protective effect against insulin release in harsh acidic conditions (pH=2.2) with only a small burst release (~15%) and sustained release at pH 6.8. Although the encapsulation process altered insulin secondary structure whilst encapsulated into PLGA NPs, insulin restored its tertiary structure once released from the NPs. Mucopenetrating heparin sulfate conjugated PLGA NPs significantly improved insulin permeability in triple co-cultured intestinal model compared to unmodified and free insulin with two and three-fold increase. Thus, they could be utilised as carriers for oral insulin delivery.

Immense Insulin Intestinal Uptake and Lymphatic Transport Using Bile Acid Conjugated Partially Uncapped Liposome.

We provide immense insulin absorption from the gastrointestinal tract, combining apical sodium-dependent bile acid transporter-mediated intestinal uptake and the lymphatic transport pathway. This strategy has proven to employ chondroitin sulfate- g-taurocholic acid coated, insulin-loaded partially uncapped liposome (IPUL-CST) for type 1 diabetes mellitus (T1DM) treatment. The loading efficiency of insulin in IPUL-CST increased significantly from 33% to 75% via the partially uncapped liposome preparation method. Moreover, the IPUL-CST revealed an improved insulin protection efficacy in GIT simulated pH and digestive enzyme conditions. The high dose of IPUL-CST in the small intestine was detected 4 h post-oral administration using ex vivo optical imaging and fluorescence intensity. The IPUL-CST exhibited significantly enhanced intestinal absorption (oral bioavailability, 34%; Tmax, 9 h) and reduced blood glucose levels for 16 h in T1DM. The results demonstrated that the new investigated IPUL-CST is a promising carrier for oral insulin delivery.

PH- and Amylase-Responsive Carboxymethyl Starch/Poly(2-isobutyl-acrylic acid) Hybrid Microgels as Effective Enteric Carriers for Oral Insulin Delivery.

Oral delivery of insulin has the potential to revolutionize diabetes care since it is regarded as a noninvasive therapeutic approach without the side effects caused by frequent subcutaneous injection. However, the insulin delivery efficiency through oral route was still limited, likely due to the chemical, enzymatic and absorption barriers. In this study, a novel type of pH- and amylase-responsive microgels as an insulin drug carrier for oral administration was developed to improve the drug delivery efficiency. The microgels were prepared via aqueous dispersion copolymerization of acrylate- grafted-carboxymethyl starch (CMS- g-AA) and 2-isobutyl-acrylic acid ( iBAA). The resulting hybrid microgels with the P iBAA contents of 13.6-45.3 wt% exhibited sharp pH-sensitivity, which was revealed by the changes in particle size of the microgels and the transmittance of the microgel aqueous solution. The accelerated decomposition of the CMS-containing microgels in response to amylase was demonstrated by chromogenic reaction and morphology change. Insulin was loaded into the microgels by swelling-diffusion method, and the insulin release from the insulin-loaded microgels in vitro was found to be triggered by pH change and addition of amylase, which was highly dependent on the microgel component. Cytotoxicity assay was performed to show the good biocompatibility of the microgels. In addition, the tests of cellular uptake by Caco-2 cells and transmembrane transport through the Caco-2 cell monolayers were carried out to confirm the intestinal absorption ability of the insulin-loaded microgels. Finally, the oral administration of insulin-loaded microgels to STZ-induced diabetic rats led to a continuous decline in the fasting blood glucose level within 2 to 4 h, and the hypoglycemic effect maintained over 6 h in vivo. The relative pharmacological availability of the insulin-loaded microgels was enhanced 23-38 times compared to free-form insulin solution through oral route. Therefore, the novel starch-based microgels may have potential as an efficient platform for oral insulin delivery.

In-vitro and in-vivo cytotoxicity and efficacy evaluation of novel glycyl-glycine and alanyl-alanine conjugates of chitosan and trimethyl chitosan nano-particles as carriers for oral insulin delivery

PurposeThe aim of this research work was to explore the possibility of providing multifunctional oral insulin delivery system by conjugating several types of dipeptides on chitosan and trimethyl chitosan to be used as drug carriers. MethodConjugates of Glycyl-glycine and alanyl-alanine of chitosan and trimethyl chitosan (on primary alcohol group of polymer located on carbon 6) were synthesized and nanoparticles containing insulin were prepared for oral delivery. Preparation conditions of nanoparticles were optimized and their performance to enhance the permeability of insulin as well as cytotoxicity of nanoparticles in Caco-2 cell line was evaluated. To evaluate the efficacy of orally administered nanoparticles, nanoparticles with the most permeability enhancing ability were studied in male Wistar rats as animal model by measuring insulin and glucose Serum levels. ResultStructural study of all the conjugates by infrared spectroscopy and nuclear magnetic resonance confirmed the successful formation of the conjugates with the desirable substitution degree. By optimizing preparation conditions, nanoparticles with expected size (157.3–197.7 nm), Zeta potential (24.35–34.37 mV), polydispersity index (0.365–0.512), entrapment efficiency (70.60–86.52%) and loading capacity (30.92–56.81%), proper morphology and desirable release pattern were obtained. Glycyl-glycine and alanyl-alanine conjugate nanoparticles of trimethyl chitosan showed 2.5–3.3 folds more effective insulin permeability in Caco-2 cell line than their chitosan counterparts. In animal model, oral administration of glycyl-glycine and alanyl-alanine conjugate nanoparticles of trimethyl chitosan demonstrated reasonable increase in Serum insulin level with relative bioavailability of 17.19% and 15.46% for glycyl-glycine and alanyl-alanine conjugate nanoparticles, respectively, and reduction in Serum glucose level compared with trimethyl chitosan nanoparticles (p < 0.05). ConclusionIt seems that glycyl-glycine and alanyl-alanine conjugate nanoparticles of trimethyl chitosan have met the aim of this research work and have been able to orally deliver insulin with more than one mechanism in animal model. Hence, they are promising candidates for further research studies.

Development of poly(hydroxyethyl methacrylate) nanogel for effective oral insulin delivery

Because of uncomfortable, painful and even deleterious effects of daily injection of insulin, extensive efforts are being made worldwide for developing noninvasive drug delivery systems, especially via the oral route. In this study, we synthesized hydroxyethyl methacrylate (HEMA) nanogel via emulsion polymerization method. The morphology and stability of the nanogel were characterized by scanning electronic microscope and dynamic light scattering. In vivo results showed the soft HEMA nanogel had longer half-live in the body circulation and exhibited almost negligible uptake by the macrophage cells as compared with blank cells. For the FITC-dextran tracking for intestinal penetration, the results indicated that the FITC-dextran in the soft nanogel penetrated faster from the inner side of the abdominal segment, which explained why the soft HEMA nanogel could promote intestinal absorption of encapsulated insulin. In vivo delivery of insulin encapsulated in the soft HEMA nanogel sustained blood glucose control for 12 h, and the overall bioavailability of administrated insulin was much higher than free insulin. Our results showed that the insulin-loaded HEMA nanogel was able to efficiently control blood glucose as a delivery system, suggesting the HEMA nanogel using emulsion polymerization could be an alternative carrier for oral insulin delivery.

Probing insulin bioactivity in oral nanoparticles produced by ultrasonication-assisted emulsification/internal gelation.

Alginate–dextran sulfate-based particles obtained by emulsification/internal gelation technology can be considered suitable carriers for oral insulin delivery. A rational study focused on the emulsification and particle recovery steps was developed in order to reduce particles to the nanosize range while keeping insulin bioactivity. There was a decrease in size when ultrasonication was used during emulsification, which was more pronounced when a cosurfactant was added. Ultrasonication add-on after particle recovery decreased aggregation and led to a narrower nanoscale particle-size distribution. Insulin encapsulation efficiency was 99.3%±0.5%, attributed to the strong pH-stabilizing electrostatic effect between insulin and nanoparticle matrix polymers. Interactions between these polymers and insulin were predicted using molecular modeling studies through quantum mechanics calculations that allowed for prediction of the interaction model. In vitro release studies indicated well-preserved integrity of nanoparticles in simulated gastric fluid. Circular dichroism spectroscopy proved conformational stability of insulin and Fourier transform infrared spectroscopy technique showed rearrangements of insulin structure during processing. Moreover, in vivo biological activity in diabetic rats revealed no statistical difference when compared to nonencapsulated insulin, demonstrating retention of insulin activity. Our results demonstrate that alginate–dextran sulfate-based nanoparticles efficiently stabilize the loaded protein structure, presenting good physical properties for oral delivery of insulin.

Chitosan-graft-PAMAM–alginate core–shell nanoparticles: a safe and promising oral insulin carrier in an animal model

Calcium chloride cross-linked PAMAM grafted chitosan–alginate core–shell nanoparticles are demonstrated as potential low-toxicity carriers for oral insulin delivery.

Efficient oral insulin delivery by dendronized chitosan: in vitro and in vivo studies

The development of efficient and bio-safe polymeric carriers for oral insulin delivery is a major thrust in biomedical research. In this paper, dendronized chitosan (DCTS) is prepared using a Michael-type addition reaction by grafting polyamidoamine (PAMAM) onto chitosan to improve its water solubility, pH sensitivity, and insulin encapsulation efficiency for enhanced bioavailability of the oral insulin. The self-assembled dendronized chitosan nanoparticles are prepared using a mild coacervation method, in which almost sub-spherically shaped nanoparticles of 85–150 nm size are produced, with an insulin encapsulation of approx. 95%. In vitro release study confirms a pH-sensitive and self-sustained release of insulin, where the oral administration of these nanoparticles exhibits a pronounced hypoglycemic effect in diabetic mice, producing a relative bioavailability of ∼9.19%. As no systemic toxicity is observed with its peroral delivery, these DCTS nanoparticles can effectively serve as a promising device in the efficient administration of oral insulin.

Retracted: Synthesis, Characterization, and In vitro Studies of PLGA–PEG Nanoparticles for Oral Insulin Delivery

Therapeutic proteins and peptides are corresponding to a major area of research in biotechnology companies and current pharmaceutical. Because of their natural instability, the enormous majority of these drugs require parentéral administration. Oral insulin delivery would be a highly attractive alternative process of administration, though it continues to be a mysterious target due to the enzymatic digestion of insulin and low levels of absorption from the gastrointestinal region. Hydrogel polymers can be considered as potential carriers for oral insulin delivery. In particular, a pH responsive hydrogel composed of PLGA-PEG has shown the ability to protect insulin from enzymes in the gastric environment and release in small intestines. However, this material has not shown similar potential for oral protein delivery of further model drugs. To date, the unique interaction between PLGA-PEG and insulin, as a potential drug for oral delivery, is not completely understood. The focus of this research is synthetization and characterization of hydrogels PLGA-PEG insulin nanoparticles and also pH sensitivity of insulin nanoparticles was investigated.

Enhanced hypoglycemic effect of biotin-modified liposomes loading insulin: effect of formulation variables, intracellular trafficking, and cytotoxicity.

Peroral protein/peptide delivery has been one of the most challenging, but encouraging topics in pharmaceutics. This article was intended to explore the potential of biotin-modified liposomes (BLPs) as oral insulin delivery carriers. By incorporating biotin-DSPE into the lipid bilayer, we prepared BLPs using reverse evaporation/sonication method. We investigated hypoglycemic effects in normal rats after oral administration of BLPs, and the possible absorption mechanism by a series of in vitro tests. The relative pharmacological bioavailability of BLPs was up to 11.04% that was as much as 5.28 folds of conventional liposomes (CLPs). The results showed that the enhanced oral absorption of insulin mainly attributed to biotin ligand-mediated endocytosis. The results provided proof of BLPs as effective carriers for oral insulin delivery.

Nanoparticles and the new era in diabetes management

Diabetes mellitus (DM) has been known to mankind for more than 2000 years. DM is a group of metabolic disorder characterized by a complete lack of insulin, a relative lack of insulin, or insulin resistance. The increase in prevalence of DM is due to three influences: lifestyle, ethnicity, and age. Current challenges in diabetes management include: optimizing the use of the already available therapies to ensure adequate glycemic, blood pressure, and lipid control and to reduce complications. At present several researches have been focusing on new management options for diabetes. Among these options the use of nanomedicine is becoming an eye catching and most promising. The aim of the present review is to provide brief overview of the applications of nanoparticles (NPs) in diabetes management. The development of improved oral insulin administration is very essential for the treatment of DM to overcome the problem of daily subcutaneous injections. In diabetic patients oral administration of insulin can be beneficial not only to alleviate the pain and trauma caused by injections, but it can also mimic the physiological fate of insulin as well. It has been found that NPs of chitosan, calcium pectinate zinc oxide, alginate, casein and different polymers have been used as a carrier for oral insulin delivery. Buccal administration of insulin with absorption enhancers showed a maximum 12% pharmacological activity. Biodegradable Polymeric NPs for parenteral insulin delivery have also been used, where the insulin matrix surrounded by the nanoporous membrane containing grafted glucose oxidase. A rise in blood glucose level triggers a change in the surrounding nanoporous membrane, resulting in biodegradation and subsequent insulin delivery. Inhalable, polymeric NP-based drug delivery systems have also been tried earlier for the treatment of tuberculosis and cardiovascular disease treatment. Such approaches can be directed toward insulin delivery through inhalable NPs. All previous studies resulted in post treatment accumulation of the NPs in skin and eyes. These drug delivery technologies are in various stages of research and development. The medical applications for nanotechnology are enormous and could give medicine, including the treatment of diabetes, an entirely new outlook.

Pluronic P85/poly(lactic acid) vesicles as novel carrier for oral insulin delivery

Poly(lactic acid)-b-Pluronic-b-poly(lactic acid) (PLA-P85-PLA) vesicles were developed as novel carrier for oral insulin delivery. PLA-P85-PLA block copolymer was synthesized by ring opening polymerization of the monomer l-lactide using Pluronic copolymer P85 as the initiator. Insulin-loaded PLA-P85-PLA vesicles were prepared by dialysis method and the mean diameter of insulin-loaded PLA-P85-PLA vesicles was determined to be 178nm. The cytotoxicity studies using human ovarian cancer cells OVCAR-3 indicate that PLA-P85-PLA block copolymer has good biocompatibility. Both in vitro and in vivo release behavior of insulin loaded in PLA-P85-PLA vesicles were studied. It was observed that insulin was released out gradually from PLA-P85-PLA vesicles and almost all insulin was released out 7.5h later. More importantly, for the oral administration of insulin-loaded PLA-P85-PLA vesicles at insulin doses of 200IU/kg, the minimum blood glucose concentration was observed in the diabetic mice test after 2.5h, which was 15% of initial glucose level. Furthermore, the blood glucose concentration increased slowly to 31.8% of initial blood glucose concentration after 10.5h and was maintained at this level for at least an additional 14h (32.5% of initial blood glucose concentration at 24.5h). These results proved that PLA-P85-PLA vesicles could be promising polymeric carriers for oral insulin delivery application due to their sustained and enhanced hypoglycemic effect.

&lt;I&gt;In Vitro&lt;/I&gt; Evaluation of Thiolated Polydimethylaminoethylmethacrylate Hydrogel Sub-Microparticles for Oral Insulin Delivery

In this investigation, novel cationic thiomer, Thiolated polydimethylaminoethylmethacrylate (PDCys) is synthesised and the feasibility of PDCys sub-microparticles as oral insulin delivery carriers is evaluated in vitro. The presence of both positive charge and thiol group in the same matrix plays a vital role in improving the paracellular permeability. Thiol groups interacts with cysteine rich subdomains via disulfide bond formation and positive charge interacts with sialic residues of mucus glycoproteins via electrostatic interaction, thereby increasing the mucoadhesivity. Cytotoxic evaluation by MTT assay shows that PDCys is nontoxic. Force and Work of adhesion of PDCys was found found to be higher than that of parent polymer. ELISA and Circular dichroism spectra confirms that PDCys retains the biological activity and conformation of insulin. Moreover, PDCys is capable of opening the tight junctions by actin and occludin filament dislocation. Furthermore, permeation of FD4 on Caco-2 cells is improved by 3.9 fold compared to the parent polymer. Preliminary studies suggest that thiolated particles can serve as potential vehicles for oral insulin delivery.

Calcium depletion-mediated protease inhibition and apical-junctional-complex disassembly via an EGTA-conjugated carrier for oral insulin delivery

Calcium (Ca2+) has a crucial role in maintaining the intestinal protease activity and in forming the apical junctional complex (AJC) that preserves epithelial barrier function. Ethylene glycol tetraacetic acid (EGTA) is a Ca2+-specific chelating agent. To maintain the concentration of this chelator in areas where enzyme inhibition and paracellular permeation enhancement are needed, this study synthesized a poly(γ-glutamic acid)–EGTA conjugate (γPGA–EGTA) to form nanoparticles (NPs) with chitosan (CS) for oral insulin delivery. The results of our molecular dynamic (MD) simulations indicate that Ca2+ ions could be specifically chelated to the nitrogen atoms, ether oxygen atoms, and carboxylate oxygen atoms in [Ca(EGTA)]2− anions. By chelating Ca2+, γPGA–EGTA conferred a significant insulin protection effect against proteases in intestinal tracts isolated from rats. Additionally, calcium depletion by γPGA–EGTA could stimulate the endocytosis of AJC components in Caco-2 cell monolayers, which led to a reversible opening of AJCs and thus increased their paracellular permeability. Single-photon emission computed tomography images performed in the biodistribution study clearly show the 123I-insulin orally delivered by CS/γPGA–EGTA NPs in the heart, aorta, renal cortex, renal pelvis and liver, which ultimately produced a significant and prolonged hypoglycemic effect in diabetic rats. The above results confirm that this γPGA–EGTA conjugate is a promising candidate for oral insulin delivery.

Novel Biodegradable and pH‐Sensitive Poly(ester amide) Microspheres for Oral Insulin Delivery

Biodegradable and pH-sensitive PEAs based on dual amino acids are designed, synthesized, and characterized. Insulin can be loaded into the PEA microspheres by a solid-in-oil-in-oil technique with high encapsulation efficiency. The feasibility of PEA microspheres as oral insulin delivery carriers is evaluated in vitro and in vivo. The hydrophobic leucine groups on PEA seem to play an important role in the pH-dependent release mechanism and cytotoxicity of PEA microspheres. Oral administration of insulin-loaded PEA microspheres to streptozotocin-induced diabetic rats at 60 IU kg(-1) is able to reduce fasting plasma glucose levels to 49.4%. These results indicate that PEA microspheres are potential new vehicles for insulin oral delivery.

Development and in-vivo evaluation of insulin-loaded chitosan phthalate microspheres for oral delivery

Novel chitosan phthalate microspheres containing insulin were prepared by emulsion cross-linking technique. The feasibility of these microspheres as oral insulin delivery carriers was evaluated. The pH-responsive release behaviour of insulin from microspheres was analysed. The ability of chitosan phthalate-insulin microspheres to enhance intestinal absorption and improve the relative pharmacological availability of insulin was investigated by monitoring the plasma glucose and insulin level of streptozotocin-induced diabetic rats after oral administration of microspheres at insulin dose of 20 IU kg(-1). In simulated gastric fluid (pH 2.0), insulin release from the microspheres was very slow. However, as the pH of the medium was changed to simulated intestinal fluid (pH 7.4), a rapid release of insulin occurred. The relative pharmacological efficacy for chitosan phthalate microspheres (18.66 +/- 3.84%) was almost four-fold higher than the efficacy of the chitosan phthalate-insulin solution administration (4.08 +/- 1.52%). Chitosan phthalate microspheres sustained the plasma glucose at pre-diabetic level for at least 16 h. These findings suggest that the microsphere is a promising carrier as oral insulin delivery system.