Colonic Delivery Research Articles

Efficient colonic drug delivery in domestic pigs employing a tablet formulation with dual control concept.

Efficient and reproducible colonic drug delivery remains elusive. The aim of this study was to demonstrate specific colonic delivery in vivo in domestic pigs with a novel tablet formulation based on a dual release control concept using 5-aminosalicylic acid (5-ASA) and caffeine as drug substances. The developed controlled colonic release (CCR) tablet formulation employs a pH-sensitive coating based on Eudragit® FS 30 D to prevent drug release in the upper gastrointestinal tract, and a xyloglucan-based matrix to inhibit drug release after coating removal in the small intestine and to allow microbiome-triggered drug release by enzymatic action in the colon. CCR tablets were administered to domestic pigs and plasma concentration data was analyzed by physiologically based pharmacokinetic modeling to estimate absorbed amounts from small and large intestine and in vivo drug release rates by model-dependent deconvolution using immediate release (IR) tablets and intravenous solutions as reference. Peak concentration times (tmax) and values (cmax) of CCR 5-ASA and caffeine tablets indicated strongly delayed drug absorption and the deduced absorbed amount as a function of time confirmed absorption overwhelmingly from the large intestine. The microbially cleaved marker molecule sulfasalazine administered alone or together with caffeine in CCR tablets reported, in combination with telemetry measurements, gastrointestinal transit times and site of absorption. Drug release from CCR tablets was inferred to take place predominantly at the site of absorption at a release rate of caffeine that was much larger in the colon than in the small intestine indicating enzymatically triggered release by the colonic microbiome. Xyloglucanase activity in rectal and cecal samples was consistent with release data and compound recovery in fecal droppings was consistent with 5-ASA bioavailability. The results provide evidence that the developed formulation can prevent premature drug release and provide targeted colonic drug delivery. Clinical relevance based on the comparability between pig and man is discussed.

Eudragit® FS Microparticles Containing Bacteriophages, Prepared by Spray-Drying for Oral Administration.

Phage therapy is recognized to be a promising alternative to fight antibiotic-resistant infections. In the quest for oral dosage forms containing bacteriophages, the utilization of colonic-release Eudragit® derivatives has shown potential in shielding bacteriophages from the challenges encountered within the gastrointestinal tract, such as fluctuating pH levels and the presence of digestive enzymes. Consequently, this study aimed to develop targeted oral delivery systems for bacteriophages, specifically focusing on colon delivery and employing Eudragit® FS30D as the excipient. The bacteriophage model used was LUZ19. An optimized formulation was established to not only preserve the activity of LUZ19 during the manufacturing process but also ensure its protection from highly acidic conditions. Flowability assessments were conducted for both capsule filling and tableting processes. Furthermore, the viability of the bacteriophages remained unaffected by the tableting process. Additionally, the release of LUZ19 from the developed system was evaluated using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME®) model. Finally, stability studies demonstrated that the powder remained stable for at least 6 months when stored at +5 °C.

Electrospinning and Electrospraying: Emerging Techniques for Probiotic Stabilization and Application.

Probiotics are beneficial for human health. However, they are vulnerable to adverse effects during processing, storage, and passage through the gastrointestinal tract, thus reducing their viability. The exploration of strategies for probiotic stabilization is essential for application and function. Electrospinning and electrospraying, two electrohydrodynamic techniques with simple, mild, and versatile characteristics, have recently attracted increased interest for encapsulating and immobilizing probiotics to improve their survivability under harsh conditions and promoting high-viability delivery in the gastrointestinal tract. This review begins with a more detailed classification of electrospinning and electrospraying, especially dry electrospraying and wet electrospraying. The feasibility of electrospinning and electrospraying in the construction of probiotic carriers, as well as the efficacy of various formulations on the stabilization and colonic delivery of probiotics, are then discussed. Meanwhile, the current application of electrospun and electrosprayed probiotic formulations is introduced. Finally, the existing limitations and future opportunities for electrohydrodynamic techniques in probiotic stabilization are proposed and analyzed. This work comprehensively explains how electrospinning and electrospraying are used to stabilize probiotics, which may aid in their development in probiotic therapy and nutrition.

Development and evaluation of bioinspired pH-responsive sericin–chitosan-based hydrogel for controlled colonic delivery of PETase: Harnessing PETase-triggered degradation of microplastics

The gravity of threats posed by microplastic pollution to the environment cannot be overestimated. Being ubiquitous in the living environment, microplastics reach humans through the food chain causing various hazardous effects. Microplastics can be effectively degraded by PETase enzymes. The current study reports, for the first time, a hydrogel-encapsulated, bioinspired colonic delivery of PETase. A free radical polymerization-assisted hydrogel system was synthesized from sericin, chitosan, and acrylic acid using N,N′-methylenebisacrylamide as a crosslinker and ammonium persulfate as an initiator. The hydrogel was characterized with FTIR, PXRD, SEM, and thermal analysis to confirm the development of a stabilized hydrogel system. The hydrogel showed 61 % encapsulation efficiency, maximum swelling, and cumulative PETase release (96 %) at pH 7.4. The mechanism of PETase release exhibited the Higuchi pattern of release with an anomalous transport mechanism. SDS-PAGE analysis confirmed the preservation of the post-release structural integrity of PETase. The released PETase exhibited concentration- and time-dependent degradation of polyethylene terephthalate in vitro. The developed hydrogel system exhibited the intended features of a stimulus-sensitive carrier system that can be efficiently used for the colonic delivery of PETase.

Comparison of 5-ASA layered or matrix pellets coated with a combination of ethylcellulose and eudragits L and s in the treatment of ulcerative colitis in rats

The aim of this study was to evaluate and optimize the combination of time and pH-dependent polymers as a single coating for the design of the colon-specific drug delivery system of 5-aminosalicylic acid (5-ASA) pellets. 5-ASA matrix pellets with a 70% drug load were prepared by the extrusion-spheronization method. The optimal coating formula which included Eudragit S (ES) + Eudragit L (EL) + Ethylcellulose (EC) was predicted for the targeted drug delivery to the colonic area by a 32 factorial design. The ratio of ES:EL:EC and coating level were considered as independent variables while the responses were the release of less than 10% of the drug within 2 h (Y1), the release of 60–70% within 10 h at pH 6.8 (Y2) and lag time of less than 1 h at pH 7.2 (Y3). Also, 5-ASA layered pellets were prepared by the powder layering of 5-ASA on nonpareils (0.4–0.6 mm) in a fluidized bed coater and then coated with the same optimum coating composition. The coated 5-ASA layered or matrix pellets were tested in a rat model of ulcerative colitis (UC) and compared with the commercial form of 5-ASA pellets (Pentasa®). The ratio of ES:EL:EC of 33:52:15 w/w at a coating level of 7% was discovered as the optimum coating for the delivery of 5-ASA matrix pellets to the colon. The coated 5-ASA pellets were spherical with uniform coating as shown by SEM and met all of our release criteria as predicted. In-vivo studies demonstrated that the optimum 5-ASA layered or matrix pellets had superior anti-inflammatory activities than Pentasa® in terms of colitis activity index (CAI), colon damage score (CDS), colon/body weight ratio and colon’s tissue enzymes of glutathione (GSH) and malondialdehyde (MDA). The optimum coating formulation showed a high potential for colonic delivery of 5-ASA layered or matrix pellets and triggered drug release based on pH and time.

Colon Delivery of Nutraceutical Ingredients by Food-Grade Polymeric Systems: An Overview of Technological Characterization and Biological Evaluation

The development of food-grade carriers based on EFSA and/or FDA-approved polymeric materials is an area of growing interest for the targeted delivery of bioactive compounds to the colon. Many nutraceuticals have shown promise in the local treatment of conditions that threaten quality of life, such as ulcerative colitis, Crohn’s disease, colorectal cancer, dysbiosis and other problems affecting the gut and colon. Nevertheless, their bioavailability is often limited due to poor solubility, rapid metabolism and low permeability, as well as undesirable local side effects. Encapsulation in carriers, which can protect the active ingredient from degradation and improve absorption and targeted administration in the colon, is one way to overcome these limitations. The technological characterization of these systems is important to assess their efficacy, safety and stability. In particular, morphology, size and surface properties influence their actions and interaction with the bio-phase. Meanwhile, encapsulation efficiency, profile and in vitro release kinetics are key parameters to assess the ability to reach the target site. This paper proposes a recent review of food-grade polymer-based systems for colorectal targeting of bioactive substances, focusing on their technological characterization and assessment of stability and biological activity, which are important in determining their full bench-to-bed potential.

Microscopic Evidence of the Behavior of pH-sensitive Food-grade Polymeric Delivery Systems

Background: Colon delivery systems are designed for the oral delivery of active compounds in the large intestine. Food-grade copolymers Eudraguard® Biotic (EUGB) and control (EUGC) have been investigated to develop colloidal systems loading natural active ingredients. Methods: In this study, we evaluated the degradation process of these matrices in simulated gastric, intestinal and colonic conditions. Microparticles made of EUGB and EUGC, alone or in combination, were loaded with the model compound resveratrol (RSV). A parallel study was performed on in vitro RSV release and SEM analysis of microparticles kept at different pH values. Results: All systems ensured a limited gastric release of RSV (below 20%), presenting only small pores on the surface of microparticles treated with simulated gastric fluid. EUGB microparticles showed the maximum release in simulated colon fluid (SCF), showing a complete dissolution of the microparticle matrix. The EUGC-based system allowed a prolonged release of RSV over time, and in SCF, it showed only partial degradation. Using mixed EUGB/EUGC matrices, a prolonged RSV release was observed along the intestinal tract. Conclusion: Overall, EUGB and EUGC copolymers were able to modulate and localize the release of entrapped cargo in the small intestine and colon. They could have interesting applications in treating bowel diseases synergistically with other therapeutic strategies.

Oral delivery of porous starch-loaded bilayer microgels for controlled drug delivery and treatment of ulcerative colitis

We prepared one type of bilayer microgels for oral administration with three effects: pH responsiveness, time lag, and colon enzyme degradation. Combined with the dual biological effects of curcumin (Cur) for reducing inflammation and promoting repair of colonic mucosal injury, targeted colonic localization and release of Cur according to the colonic microenvironment were enhanced. The inner core, derived from guar gum and low-methoxyl pectin, afforded colonic adhesion and degradation behavior; the outer layer, modified by alginate and chitosan via polyelectrolyte interaction, achieved colonic localization. The porous starch (PS)-mediated strong adsorption allowed Cur loading in inner core to achieve a multifunctional delivery system. In vitro, the formulations exhibited good bioresponses at different pH conditions, potentially delaying Cur release in the upper gastrointestinal tract. In vivo, dextran sulfate sodium-induced ulcerative colitis (UC) symptoms were significantly alleviated after oral administration, accompanied by reduced levels of inflammatory factors. The formulations facilitated colonic delivery, allowing Cur accumulation in colonic tissue. Moreover, the formulations could alter gut microbiota composition in mice. During Cur delivery, each formulation increased species richness, decreased pathogenic bacterial content, and afforded synergistic effects against UC. These PS-loaded bilayer microgels, exhibiting excellent biocompatibility, multi-bioresponsiveness, and colon targeting, could be beneficial in UC therapy, allowing development into a novel oral formulation.

Modification of Octenyl Succinic Anhydride Starch by Grafting Folic Acid and its Potential as an Oral Colonic Delivery Carrier

AbstractIn this study, an oral colonic delivery system is designed using octenyl succinic anhydride (OSA) starch grafted with folic acid (FA). OSA starch with different degrees of substitution (DS) (0.011, 0.027, and 0.039) is prepared by reacting OSA with waxy maize starch. To optimize the drug‐loading efficiency, the study choses OSA6 starch (DS = 0.027) as the substrate to prepare OSA starch grafted with FA (FA‐OSA6). Fourier transform infrared spectrum confirms the successful introduction of FA. The scanning electron micrograph indicates that the surface of the FA‐OSA starch granules is rough and uneven. Drug loading analysis shows that FA‐OSA6 starch efficiently loaded doxorubicin hydrochloride (DOX) up to 87.71%. In vitro release studies suggest that the drug‐loaded complex released only 15.8% DOX through the upper gastrointestinal tract. The cell viability assay shows that the drug‐loaded complex effectively inhibits the growth of colon cancer cells (Caco‐2). These results suggest that OSA6 starch grafted with FA has potential as an oral colonic delivery carrier for DOX.

Design and Evaluation of Controlled Onset Extended-Release System for Chronotherapeutic Delivery of Aceclofenac

The purpose of this research is to design and test Aceclofenac-containing capsule dosage type, which may be intended for the treatment of early morning stiffness and symptomatic pain relief in patients with rheumatoid arthritis. Immediate release tablets were prepared by wet granulation method using crospovidone, SSG, Lycoat, and Ludiflash. Enteric coated Microspheres were prepared using Eudragit L100 and Eudragit R100 by oil-in-oil solvent evaporation method. Both IR tablets and microspheres of aceclofenac have been tested for pre-compression, post-compression and findings find to be appropriate in our trials. Chemical reactions between the drug and the polymeric substance have been tested using FTIR. There was no disparity between the IR patterns of Aceclofenac, the physical mixture of Aceclofenac and polymers. Based on the results obtained IR tablet (AI12) containing Crospovidone as super disintegrant was considered as the optimum powder blend for fabrication of capsule system. Among the enteric coated microspheres AC9 was considered as optimized formulation for colonic drug delivery. As our aim of work is to maintain a chronotherapeutic drug delivery, now we combine the optimized IR tablets and optimized enteric coated microspheres in a single capsule. Based on the drug release studies, it shows drug released up to 20hrs and follows zero order release kinetics with non-ficikian diffusion mechanism.

Exploring the potential of quercetin/aspirin-loaded chitosan nanoparticles coated with Eudragit L100 in the treatment of induced-colorectal cancer in rats.

Growing evidence suggests quercetin and aspirin may have anticancer properties, notably in the case of colorectal cancer. The goal of this study was to create Pluronic F127 and polyethylene glycol4000 solid dispersion-loaded chitosan nanoparticles for colonic quercetin and aspirin delivery. In 1:1 polymeric stoichiometric ratio, solubility and complex formation were verified. Solid dispersion-loaded chitosan nanoparticles with a diameter of 244.45 ± 8.5nm, a surface charge of 34.1 ± 3.3mV, and encapsulation effectiveness of 76.3 ± 4.3% were generated under ideal conditions. In some cases, coating with Eudragit L100 resulted in a decrease in zeta potential and an increase in particle size. The coated formulation released the actives in a pH-dependent manner, considering their physicochemical features. Surprisingly, when compared to the actives' suspension and uncoated formulation, the coated formulation had greater anti-inflammatory efficacy, with a substantial reduction of PGE2 and IL-8 production in colonic tissues (16.9 ± 7.9ng/g tissue and 134.9 ± 10.1pg/g tissue, respectively). It also reversed most of the dimethyl hydrazine-induced histological alterations in the colon. It also demonstrated a greater reduction in TNF expression in colonic tissues. As a result, Eudragit L100-coated QT/AS-loaded chitosan nanoparticles are suggested to provide a potential platform for colonic delivery of quercetin and aspirin.

Newly developed nano-biocomposite embedded hydrogel to enhance drug loading and modulated release of anti-inflammatory drug

A newly developed iron-based nano-biocomposite (nano Fe-CNB) impregnated alginate formulation (CA) is proposed to improve drug loading and exhibit pH-responsive behavior of model anti-inflammatory drug-ibuprofen for controlled release applications. The proposed formulation is investigated with conventional β-CD addition in CA. The nano Fe-CNB-based formulations with and without β-CD, (Fe-CNB β-CD CA and Fe-CNB CA) are compared with only CA and β-CD incorporated CA formulations. The results indicate the incorporation of nano-biocomposite or β-CD into CA enhances the drug loading (>40%). However, pH-responsive controlled release behavior is observed for nano Fe-CNB based formulations only. The release studies from Fe-CNB β-CD CA indicate ∼ 45% release in stomach pH (1.2) within 2 h. In contrast, Fe-CNB CA shows ∼20% release only in stomach pH and improved release (∼49%) at colon pH (7.4). The rheology and swelling studies indicate Fe-CNB CA remains intact in stomach pH with a minimal drug release, but it disintegrates at colon pH due to charge reversal behavior of nano-biocomposite and ionization of polymeric chains. Thus, Fe-CNB CA formulation is found to be a potential candidate for targeting colon delivery, inflammatory bowel disease, and post-operative conditions.

Targeted therapeutic effects of oral magnetically driven pectin nanoparticles containing chlorogenic acid on colon cancer

Oral colonic nano-drug delivery system has attracted growing attention in treating colon cancer for their excellent characteristics. However, the unique and complex structure of the gastrointestinal tract is still an obstacle to the safe delivery of drugs targeting sites in colon tumors. Here, we designed magnetically driven dual-targeted oral colonic nanoparticles loaded with chlorogenic acid using pectin and oleic acid-modified iron oxide (Fe3O4@OA). Specific degradation of pectin by pectinase produced by colonic flora and magnetic fields applied to the colon confers specific targeting of nanoparticles to the colon. In order to overcome the challenge of preparing magnetically driven nanoparticles with small and homogeneous particle sizes by a single conventional method, we developed the combined ultrasound-emulsification technique. The average particle size of the prepared nanoparticles was 81.04 ± 1.02 nm, which showed good drug release in the simulated colonic environment. In vitro anticancer studies, the drug-loaded nanoparticles possess an obvious toxicity and apoptosis-inducing ability against cancer cells. Meanwhile, the hemolysis results demonstrated the safety of the nanoplatform (PET/CGA/Fe3O4@OA). This work holds broad prospects as a new treatment modality for colon cancer.

Control release of α-mangostin by a novel dual-polysaccharides delivery system for colitis treatment under simulated gastrointestinal conditions

The poor hydrophilia and strong cytotoxicity of α-mangostin, a major component of xanthones from pericarps of mangosteen (Garcinia Mangostana L.), restrict its application in functional food. To overcome its shortage, a series of alginate/xanthan gum or alginate/κ-carrageenan hydrogels loaded with α-mangostin were fabricated and their anti-inflammatory effects were investigated under gastrointestinal tract (GIT). With the increased concentration of polysaccharides, the release of α-mangostin was slower due to denser microstructure and hardness. The digestive profile suggested that the release of α-mangostin in 0.3% alginate/0.15% xanthan gum (0.15%X-αM), 0.3% alginate/0.2% xanthan gum (0.25%X-αM) and 0.3% alginate/0.3% κ-carrageenan (0.3%C-αM) hydrogels were faster than the others. The cytotoxicity of α-mangostin loaded in hydrogels was dramatically reduced. However, the anti-inflammatory effect of the three intestinal digestive fluids was still maintained at 71.9% ± 1.2%, 83.1% ± 2.0%, and 70.6% ± 3.3% in LPS-induced Raw246.7 cells. In addition, the three intestinal digestive fluids (50 μM of α-mangostin) showed good inhibition of inflammatory cytokines (TNF-α, IL-6, and IL-1β) and the expression of iNos and Cox-2 respectively. Given these data, xanthan gum/κ-carrageenan-alginate composite hydrogels can be a good colonic target delivery system for bioactive compounds, which can be applied in functional foods to improve human health and wellness.

Multilayered polymer coating modulates mucoadhesive and biological properties of camptothecin-loaded lipid nanocapsules

Lipid core nanocapsules (NCs) coated with multiple polymer layers were rationally designed as a potential approach for the colonic delivery of camptothecin (CPT). Chitosan (CS), hyaluronic acid (HA) and hypromellose phthalate (HP) were selected as coating materials, to modulate the mucoadhesive and permeability properties of CPT regarding the improvement of local and targeted action in the colon cancer cells. NCs were prepared by emulsification/solvent evaporation method and coated with multiple polymer layers by polyelectrolyte complexation technique. NCs exhibited spherical shape, negative zeta potential, and size ranged from 184 to 252 nm. The high efficiency of CPT incorporation (>94%) was evidenced. The ex vivo permeation assay showed that nanoencapsulation reduced the permeation rate of CPT through the intestinal mucosa by up to 3.5 times, and coating with HA and HP reduced the permeation percentage by 2 times when compared to NCs coated only with CS. The mucoadhesive capacity of NCs was demonstrated in gastric and enteric pH. Nanoencapsulation did not reduce the antiangiogenic activity of CPT and, additionally, it was observed that nanoencapsulation resulted in localized antiangiogenic action of CPT.

An Innovative Approach for Formulation of Rutin Tablets Targeted for Colon Cancer Treatment

The aim of this study was the improvement of rutin solubility along with targeting its release to colon for effective treatment of colon cancer. Five formulations of compression-coated tablets were prepared with the same core composition including rutin-polyvinyl pyrrolidone K30 solid dispersion (rutin-PVP K30 SD) but differ in being coated with either frankincense alone or different combinations of frankincense with gelatin. The superior formula was selected based on the in vitro drug release then further evaluated in terms of physical properties and in vivo performance in dogs using X-ray. Moreover, in vitro cytotoxicity of rutin, rutin-PVP K30 SD, frankincense, and a mixture of rutin-PVP K30 SD with frankincense in a ratio representing their concentrations in the selected formula was assessed against human colon cancer (HCT-116) cell lines using sulforhodamine B assay. The formula (F4) with the coat consisted of 65%w/w frankincense and 35%w/w gelatin achieved acceptable in vitro controlled drug release. In vivo X-ray in dogs confirmed that F4 tablet could remain intact in the stomach and small intestine until reaching the colon. In vitro cytotoxicity revealed that mixture of rutin-PVP K30 SD with frankincense was more effective in arresting cancer cell growth than rutin or frankincense alone. Moreover, stability studies revealed that F4 tablets were physically and chemically stable. Thus, improving rutin solubility using solid dispersion technique and formulating it into frankincense-based compression-coated (F4) tablets would be a successful approach for colonic delivery of rutin with potential of improving therapeutic efficacy.Graphical

A review on curcumin colon-targeted oral drug delivery systems for the treatment of inflammatory bowel disease.

Synthetic drugs and monoclonal antibodies are the typical treatments to combat inflammatory bowel disease (IBD). However, side effects are present when these treatments are used, and their continued application could be restricted by the high relapse rate of the disease. One potential alternative to these treatments is the use of plant-derived products. The use curcumin is one such treatment option that has seen an increase in usage in treating IBD. Curcumin is derived from a rhizome of turmeric (Curcuma longa), and the results of studies on the use of curcumin to treat IBD are promising. These studies suggest that curcumin interacts with cellular targets such as NF-κB, JAKs/STATs, MAPKs, TNF-α, IL-6, PPAR, and TRPV1 and may reduce the progression of IBD. Potentially, curcumin can be used as a therapeutic agent for patients with IBD when it reduces the incidence of clinical relapse. This review discusses the strategies utilized in designing and developing an oral colonic delivery dosage form of curcumin.

Delivery of a therapeutic antibody to the lower gastrointestinal tract for the treatment of Clostridium difficile infection (CDI)

The colonic delivery system of toxin neutralizing antibody is a promising method for treating Clostridium difficile infection (CDI) and has some advantages over the parental administration of a neutralizing antibody. However, colonic delivery of biologics presents several challenges, including instability of biologics during encapsulation into the delivery system and harsh conditions in the upper GI tract. In this work, we described a multi-particulate delivery system encapsulating a tetra-valent antibody ABAB-IgG1 with the potential to treat CDI. This work first approved that the cecum injection of ABAB-IgG1 into the lower GI tract of mice could relieve the symptoms, enhance the clinical score, and improve the survival rate of mice during CDI. Then, the antibody was spray layered onto mannitol beads and then enteric coated with pH-sensitive polymers to achieve colon-targeting release. The in vitro release of antibody from the multi-particulate system and the pH-sensitive release of antibody was monitored. The in vivo efficacy of this system was further examined and confirmed in mice and hamsters. In summary, the findings of this study should provide practical information and potential treatment options for CDI through colonic delivery of antibody therapeutics to the lower GI tract using a multi-particulate delivery system.

A NOVEL GUT-RESTRICTED ARYL HYDROCARBON RECEPTOR AGONIST WITH ACTIVITY IN THE DEXTRAN SODIUM SULFATE COLITIS MURINE MODEL

Abstract BACKGROUND Patients with inflammatory bowel disease (ulcerative colitis, Crohn’s disease) have decreased production of endogenous aryl hydrocarbon receptor (AhR) ligands in colon tissue. Activation of the AhR is a potential novel therapeutic target for ulcerative colitis and may play an important role in promoting immune homeostasis through increased IL-22 production, downregulation of proinflammatory mediators, and improved epithelial barrier function. Indigo naturalis, a botanical extract containing potent AhR agonists, has been effective for the treatment of moderate-to-severe and treatment-refractory ulcerative colitis in multiple clinical studies. The most potent AhR agonist in indigo naturalis is indirubin. OBJECTIVE The objective was to develop novel indirubin prodrugs that maximize local colonic delivery and minimize systemic exposure to reduce the risk of potential systemic side effects. RESULTS We developed a novel synthetic small molecule prodrug of indirubin that maximizes colonic exposure and minimizes systemic exposure. In a dextran sodium sulfate (DSS)-induced colitis murine model, oral administration of our lead prodrug significantly reduced Disease Activity Index and weight loss similar in magnitude to indirubin. The prodrug resulted in colon concentrations of indirubin that were over 100-fold greater than plasma concentrations, and low systemic organ exposure. CONCLUSION Azora’s novel indirubin prodrug enhanced colonic delivery of indirubin while maintaining minimal systemic exposure in a DSS colitis murine model and may reduce the risk of systemic side effects. This compound may be suitable for development as a first-in-class oral AhR agonist for the treatment of ulcerative colitis.

COLON TARGETED DELIVERY AND STABILIZATION OF MONOCLONAL ANTIBODIES FOR LOCAL TREATMENT OF INFLAMMATORY BOWEL DISEASES

Abstract In the era of biologics where IBD treatment has been transformed by potent antibody-based therapeutics, a key challenge still remains, to develop orally administered antibody medicines for chronic GI inflammation. This is primarily due to the challenges in accurate oral targeted delivery of drugs to the lower GI tract and the inherent enzymatic instability of antibodies in these harsh luminal environment. As a result, all antibody therapeutics in clinical development and on the market are available only as an injection leading to undesired adverse effects due to high systemic exposure and suboptimal therapeutic response due to insufficient antibody levels at the inflamed tissue. We have developed a suite of delivery technologies that are designed to bypass the harsher stomach and small intestinal conditions of the GI tract and achieve accurate targeted release and enzymatic stabilization of the antibody therapeutics in the colon. The delivery platform comprises of a film coating which is applied on the outside of the pill. The coating is a clinically validated polymer coating comprising of a combination of pH sensitive polymer and polysaccharide that is designed to start releasing drugs only when exposed to the correct pH and/or colonic bacteria, dramatically improving on the inconsistent colonic release profiles of solely pH-based coatings. The second component is in the core of the pill, a cocktail of amino acid-based excipients that are mixed together with the antibody and protects the drug from pancreatic and bacterial origin protease enzymes, allowing high luminal antibody concentration build-up that leads to inflamed tissue/systemic uptake through a combination of active and passive diffusion. We demonstrated in an acute DSS mouse model of colitis the effective colonic delivery and stabilization of an anti-IL6 antibody leading to superior uptake into the tissue compared IP injection. The superior tissue level demonstrated higher downregulation of target IL6. We are working to leverage the delivery technology for oral delivery of a variety of anti-inflammatory and epithelial wound healing protein modalities such as monoclonal antibodies, fusion proteins, cytokines, VHHs and peptides to allow for broad therapeutic interventions in indications such as Crohn’s disease, ulcerative colitis, pouchitis and celiac disease. Mechanism of oral colon targeted delivery of therapeutic antibodies in local treatment of inflammatory bowel disease. In-vivo PK of anti-IL6 antibody in acute DSS colitis mouse model. A. Impact of stabilization on antibody levels in the colon lumen compared to naked antibody and injected antibody. B. Impact of stabilization on colon tissue uptake. C. Effect of tissue conc. on colon tissue IL6 target downregulation.