Glucose‐Responsive Composite Microneedle Patch for Hypoglycemia‐Triggered Delivery of Native Glucagon

Insulin‐dependent patients with diabetes mellitus require multiple daily injections of exogenous insulin to combat hyperglycemia. However, administration of excess insulin can lead to hypoglycemia, a life‐threatening condition characterized by abnormally low blood glucose levels (BGLs). To prevent hypoglycemia associated with intensive insulin therapy, a “smart” composite microneedle (cMN) patch is developed, which releases native glucagon at low glucose levels. The cMN patch is composed of a photo‐crosslinked methacrylated hyaluronic acid (MeHA) microneedle array with embedded multifunctional microgels. The microgels incorporate zwitterionic moieties that stabilize loaded glucagon and phenylboronic acid moieties that provide glucose‐dependent volume change to facilitate glucagon release. Hypoglycemia‐triggered release of structurally unchanged glucagon from the cMN patch is demonstrated in vitro and in a rat model of type 1 diabetes (T1D). Transdermal application of the patch prevented insulin‐induced hypoglycemia in the diabetic rats. This work is the first demonstration of a glucose‐responsive glucagon‐delivery MN patch for the prevention of hypoglycemia, which has a tremendous potential to reduce the dangers of intensive insulin therapy and improve the quality of life of patients with diabetes and their caregivers.     

Multiresponsive Supramolecular Theranostic Nanoplatform Based on Pillar[5]arene and Diphenylboronic Acid Derivatives for Integrated Glucose Sensing and Insulin Delivery

A closed‐loop “smart” insulin delivery system with the capability to mimic pancreatic cells will be highly desirable for diabetes treatment. This study reports a multiple stimuli‐responsive insulin delivery platform based on an explicit supramolecular strategy. Self‐assembled from a well‐designed amphiphilic host–guest complex formed by pillar[5]arene and a diphenylboronic acid derivative and loaded with insulin and glucose oxidase, the obtained insulin‐GOx‐loaded supramolecular vesicles can selectively recognize glucose, accompanied by the structure disruption and efficient release of the entrapped insulin triggered by the high glucose concentration as well as the in situ generated H₂O₂ and acid microenvironment during the GOx‐promoted specific oxidation of glucose into gluconic acid. Moreover, such a “smart” supramolecular theranostic nanoplatform is able to function as both a glucose sensor and a controlled insulin delivery actuator. In vivo experiments further demonstrate that this smart supramolecular nanocarrier shows fast response to hyperglycemic circumstances and can effectively regulate the glucose levels in a mouse model of type I diabetes.     

Insulin‐Responsive Glucagon Delivery for Prevention of Hypoglycemia

Hypoglycemia, the state of abnormally low blood glucose level, is an acute complication of insulin and sulfonylurea therapy in diabetes management. Frequent insulin dosing and boluses during daily diabetes care leads to an increased risk of dangerously low glucose levels, which can cause behavioral and cognitive disturbance, seizure, coma, and even death. This study reports an insulin‐responsive glucagon delivery method based on a microneedle (MN)‐array patch for the prevention of hypoglycemia. The controlled release of glucagon is achieved in response to elevated insulin concentration by taking advantage of the specific interaction between insulin aptamer and target insulin. Integrating a painless MN‐array patch, it is demonstrated that this insulin‐triggered glucagon delivery device is able to prevent hypoglycemia following a high‐dose insulin injection in a chemically induced type 1 diabetic mouse model.     

Bioresponsive Microneedles with a Sheath Structure for H2O2 and pH Cascade‐Triggered Insulin Delivery

Self‐regulating glucose‐responsive insulin delivery systems have great potential to improve clinical outcomes and quality of life among patients with diabetes. Herein, an H₂O₂‐labile and positively charged amphiphilic diblock copolymer is synthesized, which is subsequently used to form nano‐sized complex micelles (NCs) with insulin and glucose oxidase of pH‐tunable negative charges. Both NCs are loaded into the crosslinked core of a microneedle array patch for transcutaneous delivery. The microneedle core is additionally coated with a thin sheath structure embedding H₂O₂‐scavenging enzyme to mitigate the injury of H₂O₂ toward normal tissues. The resulting microneedle patch can release insulin with rapid responsiveness under hyperglycemic conditions owing to an oxidative and acidic environment because of glucose oxidation, and can therefore effectively regulate blood glucose levels within a normal range on a chemically induced type 1 diabetic mouse model with enhanced biocompatibility.     

Blood glucose concentration control for type 1 diabetic patients: a multiple‐model strategy

In this study, a multiple‐model strategy is evaluated as an alternative closed‐loop method for subcutaneous insulin delivery in type 1 diabetes. Non‐linearities of the glucose–insulin regulatory system are considered by modelling the system around five different operating points. After conducting some identification experiments in the UVA/Padova metabolic simulator (accepted simulator by the US Food and Drug Administration (FDA)), five transfer functions are obtained for these operating points. Paying attention to some physiological facts, the control objectives such as the required settling time and permissible bounds of overshoots and undershoots are determined for any transfer functions. Then, five PID controllers are tuned to achieve these objectives and a bank of controllers is constructed. To cope with difficulties of the presence of delays in subcutaneous blood glucose (BG) measuring and in administration of insulin, a glucose‐dependent setpoint is considered as the desired trajectory for the BG concentration. The performance of the obtained closed‐loop glucose–insulin regulatory system is investigated on the in silico adult cohort of the UVA/Padova metabolic simulator. The obtained results show that the proposed multiple‐model strategy leads to a closed‐loop mechanism with limited hyperglycemia and no severe hypoglycemia.Inspec keywords: blood, patient diagnosis, medical control systems, biochemistry, three‐term control, closed loop systems, diseases, patient treatment, drugs, sugarOther keywords: blood glucose concentration control, type 1 diabetic patients, multiple‐model strategy, alternative closed‐loop method, subcutaneous insulin delivery, type 1 diabetes, transfer functions, control objectives, PID controllers, subcutaneous blood glucose measuring, glucose‐dependent setpoint, closed‐loop glucose–insulin regulatory system, closed‐loop mechanism     

Ultrasound-triggered noninvasive regulation of blood glucose levels using microgels integrated with insulin nanocapsules

Diabetes is a serious public health problem affecting 422 million people worldwide.Traditional diabetes management often requires multiple daily insulin injections,associated with pain and inadequate glycemia control.Herein,we have developed an ultrasound-triggered insulin delivery system capable of pulsatile insulin release that can provide both long-term sustained and fast on-demand responses.In this system,insulin-loaded poly(lactic-co-glycolic acid) (PLGA) nanocapsules are encapsulated within chitosan microgels.The encapsulated insulin in nanocapsules can passively diffuse from the nanoparticle but remain restricted within the microgel.Upon ultrasound treatment,the stored insulin in microgels can be rapidly released to regulate blood glucose levels.In a chemically-induced type 1 diabetic mouse model,we demonstrated that this system,when activated by 30 s ultrasound administration,could effectively achieve glycemic control for up to one week in a noninvasive,localized,and pulsatile manner.     

Robust multi‐objective blood glucose control in Type‐1 diabetic patient

In this study, an automatic robust multi‐objective controller has been proposed for blood glucose (BG) regulation in Type‐1 Diabetic Mellitus (T1DM) patient through subcutaneous route. The main objective of this work is to control the BG level in T1DM patient in the presence of unannounced meal disturbances and other external noises with a minimum amount of insulin infusion rate. The multi‐objective output‐feedback controller with H_∞, H₂ and pole‐placement constraints has been designed using linear matrix inequality technique. The designed controller for subcutaneous insulin delivery was tested on in silico adult and adolescent subjects of UVa/Padova T1DM metabolic simulator. The experimental results show that the closed‐loop system tracks the reference BG level very well and does not show any hypoglycaemia effect even during the long gap of a meal at night both for in silico adults and adolescent. In the presence of 50 gm meal disturbance, average adult experience normoglycaemia 92% of the total simulation time and hypoglycaemia 0% of total simulation time. The robustness of the controller has been tested in the presence of irregular meals and insulin pump noise and error. The controller yielded robust performance with a lesser amount of insulin infusion rate than the other designed controllers reported earlier.Inspec keywords: robust control, patient treatment, diseases, closed loop systems, patient monitoring, biochemistry, medical control systems, blood, organic compoundsOther keywords: robust multiobjective blood glucose control, automatic robust multiobjective controller, blood glucose regulation, Type‐1 Diabetic Mellitus patient, BG level, T1DM patient, insulin infusion rate, multiobjective output‐feedback controller, pole‐placement constraints, linear matrix inequality technique, subcutaneous insulin delivery, total simulation time, insulin pump noise, adolescent subjects, meal disturbance, normoglycaemia 92, in silico adults, UVa‐Padova T1DM metabolic simulator, closed‐loop system, hypoglycaemia effect     

Light‐Triggered Biomimetic Nanoerythrocyte for Tumor‐Targeted Lung Metastatic Combination Therapy of Malignant Melanoma

Red blood cell (RBC) membrane‐cloaked nanoparticles, reserving the intact cell membrane structure and membrane protein, can gain excellent cell‐specific functions such as long blood circulation and immune escape, providing a promising therapy nanoplatform for drug delivery. Herein, a novel RBC membrane biomimetic combination therapeutic system with tumor targeting ability is constructed by embedding bovine serum albumin (BSA) encapsulated with 1,2‐diaminocyclohexane‐platinum (II) (DACHPt) and indocyanine green (ICG) in the targeting peptide‐modified erythrocyte membrane (R‐RBC@BPtI) for enhancing tumor internalization and synergetic chemophototherapy. R‐RBC@BPtI displays excellent stability and high encapsulation efficiency with multiple cores enveloped in the membrane. Benefited from the stealth functionality and targeting modification of erythrocyte membranes, R‐RBC@BPtI can significantly promote tumor targeting and cellular uptake. Under the near‐infrared laser stimuli, R‐RBC@BPtI presents remarkable instability by singlet oxygen and heat‐mediated cleavage so as to trigger effective drug release, thereby achieving deep penetration and accumulation of DACHPt and ROS in the tumor site. Consequently, R‐RBC@BPtI with tumor‐specific targeting ability accomplishes remarkable ablation of tumors and suppressed lung metastasis in vivo by photothermal and chemotherapy combined ablation under phototriggering. This research provides a novel strategy of targeted biomimetic nanoplatforms for combined cancer chemotherapy–phototherapy.     

Nanomechanics of Extracellular Vesicles Reveals Vesiculation Pathways

Extracellular vesicles (EVs) are emerging as important mediators of cell–cell communication as well as potential disease biomarkers and drug delivery vehicles. However, the mechanical properties of these vesicles are largely unknown, and processes leading to microvesicle‐shedding from the plasma membrane are not well understood. Here an in depth atomic force microscopy force spectroscopy study of the mechanical properties of natural EVs is presented. It is found that several natural vesicles of different origin have a different composition of lipids and proteins, but similar mechanical properties. However, vesicles generated by red blood cells (RBC) at different temperatures/incubation times are different mechanically. Quantifying the lipid content of EVs reveals that their stiffness decreases with the increase in their protein/lipid ratio. Further, by maintaining RBC at “extreme” nonphysiological conditions, the cells are pushed to utilize different vesicle generation pathways. It is found that RBCs can generate protein‐rich soft vesicles, possibly driven by protein aggregation, and low membrane–protein content stiff vesicles, likely driven by cytoskeleton‐induced buckling. Since similar cortical cytoskeleton to that of the RBC exists on the membranes of most mammalian cells, our findings help advancing the understanding of the fundamental process of vesicle generation.     

Surface‐Engineering of Red Blood Cells as Artificial Antigen Presenting Cells Promising for Cancer Immunotherapy

The development of artificial antigen presenting cells (aAPCs) to mimic the functions of APCs such as dendritic cells (DCs) to stimulate T cells and induce antitumor immune responses has attracted substantial interests in cancer immunotherapy. In this work, a unique red blood cell (RBC)‐based aAPC system is designed by engineering antigen peptide‐loaded major histocompatibility complex‐I and CD28 activation antibody on RBC surface, which are further tethered with interleukin‐2 (IL2) as a proliferation and differentiation signal. Such RBC‐based aAPC‐IL2 (R‐aAPC‐IL2) can not only provide a flexible cell surface with appropriate biophysical parameters, but also mimic the cytokine paracrine delivery. Similar to the functions of matured DCs, the R‐aAPC‐IL2 cells can facilitate the proliferation of antigen‐specific CD8+ T cells and increase the secretion of inflammatory cytokines. As a proof‐of‐concept, we treated splenocytes from C57 mice with R‐aAPC‐IL2 and discovered those splenocytes induced significant cancer‐cell‐specific lysis, implying that the R‐aAPC‐IL2 were able to re‐educate T cells and induce adoptive immune response. This work thus presents a novel RBC‐based aAPC system which can mimic the functions of antigen presenting DCs to activate T cells, promising for applications in adoptive T cell transfer or even in direct activation of circulating T cells for cancer immunotherapy.     

Erythrocyte–Platelet Hybrid Membrane Coating for Enhanced Nanoparticle Functionalization

Cell‐membrane‐coated nanoparticles have recently been studied extensively for their biological compatibility, retention of cellular properties, and adaptability to a variety of therapeutic and imaging applications. This class of nanoparticles, which has been fabricated with a variety of cell membrane coatings, including those derived from red blood cells (RBCs), platelets, white blood cells, cancer cells, and bacteria, exhibit properties that are characteristic of the source cell. In this study, a new type of biological coating is created by fusing membrane material from two different cells, providing a facile method for further enhancing nanoparticle functionality. As a proof of concept, the development of dual‐membrane‐coated nanoparticles from the fused RBC membrane and platelet membrane is demonstrated. The resulting particles, termed RBC–platelet hybrid membrane‐coated nanoparticles ([RBC‐P]NPs), are thoroughly characterized, and it is shown that they carry properties of both source cells. Further, the [RBC‐P]NP platform exhibits long circulation and suitability for further in vivo exploration. The reported strategy opens the door for the creation of biocompatible, custom‐tailored biomimetic nanoparticles with varying hybrid functionalities, which may be used to overcome the limitations of current nanoparticle‐based therapeutic and imaging platforms.     

Engineered Multifunctional Albumin‐Decorated Porous Silicon Nanoparticles for FcRn Translocation of Insulin

The last decade has seen remarkable advances in the development of drug delivery systems as alternative to parenteral injection‐based delivery of insulin. Neonatal Fc receptor (FcRn)‐mediated transcytosis has been recently proposed as a strategy to increase the transport of drugs across the intestinal epithelium. FcRn‐targeted nanoparticles (NPs) could hijack the FcRn transcytotic pathway and cross the epithelial cell layer. In this study, a novel nanoparticulate system for insulin delivery based on porous silicon NPs is proposed. After surface conjugation with albumin and loading with insulin, the NPs are encapsulated into a pH‐responsive polymeric particle by nanoprecipitation. The developed NP formulation shows controlled size and homogeneous size distribution. Transmission electron microscopy (TEM) images show successful encapsulation of the NPs into pH‐sensitive polymeric particles. No insulin release is detected at acidic conditions, but a controlled release profile is observed at intestinal pH. Toxicity studies show high compatibility of the NPs with intestinal cells. In vitro insulin permeation across the intestinal epithelium shows approximately fivefold increase when insulin is loaded into FcRn‐targeted NPs. Overall, these FcRn‐targeted NPs offer a toolbox in the development of targeted therapies for oral delivery of insulin.     

Effect of recombinant human erythropoietin on insulin resistance in hemodialysis patients

Insulin resistance is a characteristic feature of uremia. Insulin resistance and concomitant hyperinsulinemia are present irrespective of the type of renal disease. Treatment with recombinant human erythropoietin (rHuEPO) was said to be associated with improvement in insulin sensitivity in uremic patients. The aim of this study was to compare insulin resistance in adult uremic hemodialysis (HD) patients including diabetic patients treated with or without rHuEPO. A total of 59 HD patients were studied, patients were divided into 2 groups of subjects: 30 HD patients on regular rHuEPO treatment (group A), and 29 HD patients not receiving rHuEPO (group B) diabetic patients were not excluded. Full medical history and clinical examination, hematological parameters, lipid profile, serum albumin, parathyroid horomone, Kt/V, fasting glucose, and insulin levels were measured in all subjects. Homeostasis Model Assessment of Insulin Resistance (HOMA‐IR) was used to compare insulin resistance. The results of this study showed that the mean insulin level of HD patients treated with rHuEPO (group A) (17.5 ± 10.6 μU/mL) was significantly lower than patients without rHuEPO (group B) (28.8 ± 7.7 μU/mL), (P<0.001). Homeostasis Model Assessment of Insulin Resistance levels in group A were significantly lower than in group B (3.8 ± 2.97, 7.98 ± 4.9, respectively, P<0.001). Insulin resistance reflected by HOMA‐IR levels among diabetic patients in group A was significantly lower than among diabetic patients in group B (3.9 ± 3.2, 9.4 ± 7.2, respectively, P<0.001). Also, HOMA‐IR levels among nondiabetic patients in group A were significantly lower than among nondiabetic patients in group B (3.7 ± 2.85, 6.9 ± 1.43, respectively, P<0.01). We found a statistically significant negative correlation between duration of erythropoietin treatment, fasting blood glucose, insulin levels, and insulin resistance (r=?0.62, ?0.71, and ?0.57, P<0.001). Patients treated with rHuEPO showed less insulin resistance compared with patients not treated with rHuEPO in diabetic and nondiabetic patients and, duration of erythropoietin treatment is negatively correlated with insulin levels and insulin resistance in HD patients.     

A Quantitative Study of the Intracellular Dynamics of Fluorescently Labelled Glyco‐Gold Nanoparticles via Fluorescence Correlation Spectroscopy

The dynamic behaviour of gold nanoparticles functionalised with glucose (Glc‐Au NPs) has been studied here by means of fluorescence correlation spectroscopy (FCS). Meaningful data on the state of aggregation and dynamics of Glc‐Au NPs fluorescently‐labelled with HiLyte Fluor647 (Glc‐Au‐Hi NPs) in the intracellular environment were obtained. Moreover, the work presented here shows that FCS can be used to visualise the presence of single NPs or NP aggregates following uptake and to estimate, locally, NP concentrations within the cell. FCS measurements become possible after applying a “prebleaching” methodology, when the immobile NP fraction has been effectively removed and thus significant FCS data has been recorded. In this study, Glc‐Au‐Hi NPs have been incubated with HepG2 cells and their diffusion time in the intracellular environment has been measured and compared with their diffusion value in water and cell media.     

Bioinspired Diselenide‐Bridged Mesoporous Silica Nanoparticles for Dual‐Responsive Protein Delivery

Controlled delivery of protein therapeutics remains a challenge. Here, the inclusion of diselenide‐bond‐containing organosilica moieties into the framework of silica to fabricate biodegradable mesoporous silica nanoparticles (MSNs) with oxidative and redox dual‐responsiveness is reported. These diselenide‐bridged MSNs can encapsulate cytotoxic RNase A into the 8–10 nm internal pores via electrostatic interaction and release the payload via a matrix‐degradation controlled mechanism upon exposure to oxidative or redox conditions. After surface cloaking with cancer‐cell‐derived membrane fragments, these bioinspired RNase A‐loaded MSNs exhibit homologous targeting and immune‐invasion characteristics inherited from the source cancer cells. The efficient in vitro and in vivo anti‐cancer performance, which includes increased blood circulation time and enhanced tumor accumulation along with low toxicity, suggests that these cell‐membrane‐coated, dual‐responsive degradable MSNs represent a promising platform for the delivery of bio‐macromolecules such as protein and nucleic acid therapeutics.     

Enhancement of absorption of insulin-loaded polyisobutylcyanoacrylate nanospheres by sodium cholate after oral and subcutaneous administration in diabetic rats.

Polyisobutylcyanoacrylate (PIBCA) nanospheres were employed as biodegradable polymeric carriers for oral (p.o.) and subcutaneous (s.c.) delivery of insulin. The polymerization technique used was able to hold 65%-95% of insulin added 30 min after initiation of polymerization. The percentage drug loading was monomer concentration dependent. Insulin adsorption to the nanospheres was measured by radioimmunoassay. Although Pluronic F68 (0.5%) did not significantly alter the in vitro insulin degradation half-life T50%, sodium cholate (0.5%) increased the degradation T50% of insulin by 56% (from 13.6 +/- 1.6 to 22.1 +/- 2 min). This study also investigated the in vivo performance of insulin-loaded PIBCA in aqueous suspension with or without sodium cholate (0.5%) and Pluronic F68 (0.5%) surfactants after oral and subcutaneous administration to alloxan-induced diabetic rats. Insulin absorption was evaluated by its hypoglycemic effect. Insulin associated with PIBCA nanospheres retains its biological activity up to 15 h and 24 h after oral and subcutaneous administrations, respectively. Administered orally insulin-loaded (75 U/kg) nanospheres, in the presence of surfactants, significantly reduced the mean blood glucose level from 392 +/- 32 to 80 +/- 13 mg/dl within 2 h and maintained it at 100 mg/dl or less for more than 8 h. On the other hand, the subcutaneous administration of insulin-loaded (25 U/kg) nanospheres significantly decreased the blood glucose level from 406 +/- 33 to 88.5 +/- 12.8 mg/dl within 1 h, and the lowered glucose level was maintained at 100 mg/dl or less for more than 12 h; it returned to its initial value 24 h after administration. Insulin-loaded nanospheres with surfactants showed significant (P < .05) pharmacological availability (PA%) of 37.6% +/- 3.7% and 65.2% +/- 2.7% after oral and subcutaneous dosages, respectively. The existence of surfactants with PIBCA nanospheres improved the oral PA% by 49.2%. These findings suggest that the developed PIBCA, in the presence of surfactants, would be useful not only in improving insulin gastrointestinal absorption, but also in sustaining its systemic action by lowering the blood glucose to an acceptable level.     

Hierarchical Tumor Microenvironment‐Responsive Nanomedicine for Programmed Delivery of Chemotherapeutics

Nanomedicines have been demonstrated to have passive or active tumor targeting behaviors, which are promising for cancer chemotherapy. However, most nanomedicines still suffer from a suboptimal targeting effect and drug leakage, resulting in unsatisfactory treatment outcome. Herein, a hierarchical responsive nanomedicine (HRNM) is developed for programmed delivery of chemotherapeutics. The HRNMs are prepared via the self‐assembly of cyclic Arg‐Gly‐Asp (RGD) peptide conjugated triblock copolymer, poly(2‐(hexamethyleneimino)ethyl methacrylate)‐poly(oligo‐(ethylene glycol) monomethyl ether methacrylate)‐poly[reduction‐responsive camptothecin] (PC7A‐POEG‐PssCPT). In blood circulation, the RGD peptides are shielded by the POEG coating; therefore, the nanosized HRNMs can achieve effective tumor accumulation through passive targeting. Once the HRNMs reach a tumor site, due to the hydrophobic‐to‐hydrophilic conversion of PC7A chains induced by the acidic tumor microenvironment, the RGD peptides will be exposed for enhanced tumor retention and cellular internalization. Moreover, in response to the glutathione inside cells, active CPT drugs will be released rapidly for chemotherapy. The in vitro and in vivo results confirm effective tumor targeting, potent antitumor effect, and reduced systemic toxicity of the HRNMs. This HRNM is promising for enhanced chemotherapeutic delivery.     

Free‐Blockage Mesoporous Anticancer Nanoparticles Based on ROS‐Responsive Wetting Behavior of Nanopores

To achieve an excellent delivery effect of drug, stimuli‐responsive nano “gate” with physical blockage units is usually constructed on the surface of the mesoporous silica nanocarriers (MSNs). In nature, the aquaporins in cell membrane can control the transport of water molecules by regulating the channel wettability, which is resulted from the conformational change of amino acids in the channel. Inspired by this phonomenon, herein a new concept of free‐blockage controlled release system is proposed, which is achieved by controlling the wettability of the internal surface of nanopores on MSNs. Such a new system is different from the physical‐blockage controlled release system, which bypasses the use of nano “gate” and overcomes the limitations of traditional physical blockage system. Moreover, further studies have shown that the system can selectively release the entrapped doxorubicin in human breast adenocarcinoma (MCF‐7) cells triggered by intracellular reactive oxygen species (ROS) but not in normalhuman umbilical vein endothelial cells (HUVECs) containing ROS with low levels. The wettability‐determined free‐blockage controlled release system is simple and effective, and it can also be triggered by intracellular biological stimuli, which provides a new approach for the future practical application of drug delivery and cancer therapy.     

Effects of Gastrointestinal Administration of Human Insulin and a Human Insulin-Deae-Dextran Complex Entrapped in Different Compound Liposomes on Blood Glucose in Rats

The effects of gastrointestinal administration (oral, in duodenum and colon) of human insulin and a human insulin-DEAE (diethylaminoethyl) dextran complex entrapped in different compound liposomes in comparing to human insulin alone given subcutaneously on blood glucose level of normal and STZ-diabetic rats were investigated. The liposomes were prepared from a hydrogenated soy lecithin (Epikuron, E 200 H) and by a high pressure homogenization procedure. Samples were lyophilized and reconstituted in 0.067 M phosphate buffer, pH 7.4 before application. The complexed insulin (0.25 and 0.5 IU/kg insulin) showed no diffrences in blood glucose lowering profiles from the free insulin when both were administered intravenously in normal rats. When given orally, the complex (30 and 60 IU/kg Insulin) entrapped in positive liposomes (E 200 H/cholesterol/stearylamine = 7:2:1, in a molar ratio) indicated no effects in STZ rats. However, this complex liposome (6.0 IU/kg insulin) gave a retention effect of blood glucose lowering as % initial level of about 12% after 5 hours when injected in duodenum and showed two maximum effects of 21 and 22% at 100 and 300 minutes respectively when administered in colon of normal rats. When the free insulin entrapped in positive liposomes was given in duodenum In normal rats, the maximum effect of blood glucose lowering of 10% was observed at 2 hours (6 IU/kg insulin) and 1 hour (12 IU/kg insulin). For the free insulin (12 IU/kg) entrapped in other liposome systems given in duodenum of normal rats, both negative (E 200 H/cholesterol/dlcetyl phosphate=7:2:1, in a molar ratio) and neutral (E 200 H/cholesterol=1:1, in a molar ratio) liposomes indicated the maximum effect of about 30% at 120 minutes. Both cholesterol rich positive (E 200 H/cholesterol/stearylamlne=7:7:1, in a molar ratio) and negative (E 200 H/cholesterol/dicetyl phosphate=7:7:1, in a molar ratio) liposomes showed not only a maximum effect of about 20% at 2 hours, but also a retention of glucose lowering of 20% after 7 hours as well. This study suggested that a development of human Insulin by complexlng with the DEAE-dextran polymer and/or entrapping in liposomes, as a drug delivery system in duodenum and colon, is possible.     

Dual pH/ROS‐Responsive Nanoplatform with Deep Tumor Penetration and Self‐Amplified Drug Release for Enhancing Tumor Chemotherapeutic Efficacy

Poor deep tumor penetration and incomplete intracellular drug release remain challenges for antitumor nanomedicine application in clinical settings. Herein, a nanomedicine (RLPA‐NPs) is developed that can achieve prolonged blood circulation, deep tumor penetration, active‐targeting of cancer cells, endosome/lysosome escape, and intracellular selectivity self‐amplified drug release for effective drug delivery. The RLPA‐NPs are constructed by encapsulation of a pH‐sensitive polymer octadecylamine‐poly(aspartate‐1‐(3‐aminopropyl) imidazole) (OA‐P(Asp‐API)) and a ROS‐generation agent, β‐Lapachone (Lap), in micelles assembled by the tumor‐penetration peptide internalizing RGD (iRGD)‐modified ROS‐responsive paclitaxel (PTX)‐prodrug. iRGD could promote RLPA‐NPs penetration into deep tumor tissue, and specific targeting to cancer cells. After internalization by cancer cells through receptor‐mediated endocytosis, OA‐P(Asp‐API) can rapidly protonate in the endosome's acidic environment, resulting in RLPA‐NPs escape from the endosome through the “proton sponge effect”. At the same time, the RLPA‐NPs micelle disassembles, releasing Lap and PTX‐prodrug. Subsequently, the released Lap could generate ROS, consequently amplifying and accelerating PTX release to kill tumor cells. The in vitro and in vivo studies demonstrated that RLPA‐NPs can significantly improve the therapeutic effect compared to control groups. Therefore, RLPA‐NPs are a promising nanoplatform for overcoming multiple physiological and pathological barriers to enhance drug delivery.