Novel polyurethane based particulate formulations of infliximab reduce inflammation in DSS induced murine model of colitis – A preliminary study
Graphical abstract
Introduction
Inflammatory bowel disease (IBD) comprising of ulcerative colitis (UC) and crohns disease, is a chronic progressive inflammatory condition of the gastro-intestinal (GI) mucosa, characterised by varying degrees of intestinal injury (Ungaro et al., 2019). It is a chronic relapsing and remitting condition with rising incidence worldwide, with 6.8 million cases globally in 2017 (Alatab et al., 2020). If not optimally treated, IBD can lead to serious complications and disability, and is also a risk factor for developing GI cancers (Colombel et al., 2011). Clinical signs and symptoms of UC include weight loss, passing blood with loose stools. This can lead of loss in education and reduced productivity at work, resulting in economic burden (Burisch et al., 2013, Ungaro et al., 2019).
In mild to moderate cases of UC, treatment is initiated with conventional small molecule comprising of immunosuppressive agents such as aminosalicylates which mainly provide symptomatic relief, and do not induce repair of the inflammatory lesions in IBD (Colombel et al., 2020, Colombel et al., 2011). Consequently, disease progression ultimately leads to surgical intervention in 46.9% of crohns disease and 15.6% of ulcerative colitis patients, within 10 years of diagnosis (Frolkis et al., 2013).
Non-responders to conventional therapy are then switched to therapeutics for the treatment of moderate to severe colitis which include: anti-TNFα monoclonal antibody (mAbs): infliximab (Remicade®, FDA approval in 2006), adalimumab (Humira®, FDA approval in 2012), golimumab (Simponi®, FDA approval in 2013), certolizumab pegol (under Phase 2 clinical trial), anti-integrin α 4β 7 mAb: vedolizumab (Entyvio®, FDA approval in 2014), anti-IL −12 / –23 mAb: ustekinumab (Stelara®, FDA approval in 2019), small molecule targeting janus kinase (JAK) signaling: tofacitinib (Xeljanz®, FDA approval in 2018) and sphingosine-1-phosphate (S1P) receptor: ozanimod (being considered for FDA approval) (Binienda et al., 2020, Hazel and O’Connor, 2020).
In the last decade there has been a paradigm shift in the goal of UC treatment, which is to initiate and maintain clinical remission thus allowing mucosal healing, a key goal to reduction in hospitalisation and surgery (Colombel et al., 2020, Sandborn et al., 2009).
Biological therapies such as anti-TNFα monoclonal antibodies mAbs, particularly infliximab (INF) have been reported to achieve this goal and have shown good clinical benefits in IBD, improving quality of life (Colombel et al., 2011, Griffiths et al., 2020, Nurbhai et al., 2019, Singh et al., 2018, Troncone et al., 2020). However, parenteral (intravenous or subcutaneous) administration of mAbs is associated with poor patient compliance and life threatening side effects including risk of serious infections, due to high dose related long term systemic immune suppression (Nurbhai et al., 2019). In addition, mAbs are associated with non-drug costs including outpatient appointments and administration costs, adding to the economic burden of the healthcare system (Cronin et al., 2019).
Consequently, orally administered gut-selective treatment with high intestinal exposure exhibiting local effect and minimal systemic exposure and side-effects is desirable (Bhol et al., 2013, Tambuwala et al., 2015). INF stability has been measured and been found to be reasonably stable in the colon (Yadav et al., 2016). The desire to attend to this current unmet clinical need is increasing, and is reflected in recent studies surrounding development of novel oral anti-TNFα and evaluation of its efficacy in preclinical models (Almon et al., 2021).
Various experimental models of colitis have been reported in the literature, however no single model replicates all the aspects and features of human UC. Chemically induced model of colitis is most commonly used due to advantages of rapid onset of inflammation and reproducibility (Wirtz et al., 2017).
In 2,4,6-trinitro-benzene sulfonic acid (TNBS) and oxazolone induced colitis model, intrarectal administration of TNBS or Oxazolone results in T-cell mediated immunity against haptenised luminal antigens and microbiodata derived proteins. TNBS induced colitis model have been reported to exhibit many histopathological features similar to Crohns disease (Alex et al., 2009, Wirtz et al., 2017), whereas oxazolone has been reported to resemble many disease features of human UC. It induces colitis via elevated interleukins (IL) production, therefore Oxazolone induced colitis model has been used to evaluate efficacy of anti-IL therapeutics (Kasaian et al., 2014, Wirtz et al., 2017).
On other hand in DSS model, colitis is induced by oral administration of DSS dissolved in drinking water, available ad libitum. DSS is believed to induce colitis by disrupting intestinal epithelial barrier, resulting in increased permeability and allowing entry of luminal antigens to the underlying mucosal immune system, resulting in rapid and profound inflammatory response and further damage to the intestinal mucosa (Wirtz et al., 2017). DSS induced model of colitis resembles many clinical characteristics similar to human UC (Alex et al., 2009). It is easy to generate and is highly reproducible, therefore it has been used extensively to investigate the potential of therapeutics and novel drug delivery system to reduce inflammation in UC (Beloqui et al., 2014, Naeem et al., 2015, Xiao et al., 2017).
Pathophysiological changes in UC offers therapeutic potential as it allows targeted delivery of polymeric particulate formulations (NPs) by the virtue of its preferential accumulation at the inflamed site via electrostatic interactions. This potentially promotes its residence time, allowing translocation of NPs across the compromised intestinal barrier to the underlying layer of the intestinal mucosa, the site of inflammation. This also prevents its rapid elimination by diarrhoea, a common symptom in IBD (Collnot et al., 2012, Maisel et al., 2015, Schmidt et al., 2013).
Numerous authors have successfully explored this therapeutic potential by engineering polymeric particulate formulations as carriers for targeted delivery of conventional small molecules. Ali et al (Ali et al., 2014) and Xiao et al (Xiao et al., 2015) reported higher therapeutic efficacy of budesonide and curcumin based PLGA polymeric particles in DSS induced murine model of colitis by virtue of maximized local drug concentrations at inflamed sites, while minimizing any systemic side effects. In first in vivo study in IBD human patients, rectally administered non-functionalized polymeric PLGA particles showed accumulation and uptake in intestinal ulcerous lesions (Schmidt et al., 2013).
INF has been reported to be effective in ameliorating the severity of colitis in DSS induced murine model of colitis, administered via IV and rectal route (Lopetuso et al., 2013). Limited studies have been carried out in engineering particulate formulations designed for oral delivery of anti-TNFα mAbs to treat inflammation in IBD. Recently Kim et al (Kim et al., 2020) and Wang et al (Wang et al., 2020) reported higher therapeutic efficacy of orally administered INF loaded coated/uncoated liposomal nanocomposites particles and natural polyphenol tannic acid/PEG particles, respectively in DSS induced murine model of colitis, compared to INF suspensions.
In our recently published work, we compared novel polyesterurethane (PU) biomaterial with conventional PLGA and PCL as polymeric drug carrier platform and observed that novel INF loaded PU (INF-PU) based particulate formulations (NPs) showed a rapid rate and extent of recovery of the epithelial barrier function in an in-vitro inflamed epithelial cell culture model (Pabari et al., 2019). This was related to the higher cellular interaction, uptake and permeability of these novel PU based polymeric NPs (Mattu et al., 2013, Pabari et al., 2019).
Hence in this work we investigated the potential of these novel polymeric biomaterial of PU, and its PEGylated (PU-PEG) form as carriers of INF to reduce inflammation in a moderate and severe DSS induced murine model of colitis. Differential localisation of the fluorescently labelled PU and PU-PEG NPs in healthy and colitis models was also studied.
Section snippets
Formulation and characterisation of infliximab-loaded particulate formulations
INF-loaded particles were formulated by solvent dispersion technique as outlined by us previously (Pabari et al., 2019). Polymer, PU or PU-PEG was dissolved at 10 mg/ml or 20 mg/ml in acetone, and added dropwise to aqueous Tween® 80 solution at 2% w/v or 0.02% w/v, while stirring at 480 rpm. This mixture was stirred overnight to allow complete solvent evaporation, then washed and incubated with INF solution (1 mg/ml in sterile water for injection) for 2 h at 4 °C, under mild stirring. This was
Influence of increasing DSS concentrations on the severity of colitis induced
In this study DSS at increasing concentrations from 0 to 2.5 %w/v was administered in drinking water from day 0 to day 7, and the severity of colitis induced was compared. Three major clinical parameters of colitis i.e. degree of weight loss, alterations in stool consistency and blood in the stools determined by Hemdetect kit was recorded daily and calculated as disease activity index (DAI) score to determine severity of colitis induced.
Clinical signs of colitis were observed in all the DSS
Discussion
In IBD, a complex interaction between genetic factors and environmental stress is believed to cause impairment in epithelial barrier integrity, allowing entry of luminal toxins and microbes to the underlying intestinal mucosa. This results in abnormal over activation of intestinal mucosal immune responses, and secretion of pro-inflammatory cytokines causing mucosal damage. Excessive secretion of pro-inflammatory cytokine, particularly TNFα, both soluble and membrane-bound forms are reported to
Conclusion
In this study INF loaded PU and PU-PEG particulate formulations adhered to the inflamed mucosa, allowing targeted delivery of INF, anti-TNFα mAb to the site of inflammation in DSS induced murine model of colitis. This enabled local inhibition of TNFα driven pathways of inflammation at its source, reducing inflammation in both moderate and severe colitis model, and increasing therapeutic potential of INF in bringing mucosal healing, a key goal of IBD therapy.
These promising preliminary results
Author contribution
Ritesh M. Pabari: Conceptualization, Methodology, Investigation, Visualization, Formal analysis, Funding acquisition. Murtaza M. Tambuwala: Conceptualization, Methodology, Formal analysis, Supervision, Writing - review & editing (including histological scoring). Natalia Lajczak-McGinley: Methodology. Alaa Aljabali: Formal analysis, Writing - review & editing (including histological scoring). Brian P. Kirby: Supervision. Stephen Keely: Supervision. Zebunissau Ramtoola: Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
The authors gratefully acknowledge research funding from the Irish Research Council (IRC) awarded to Ritesh M. Pabari (GOIPD/2014/403). Authors would like to thank Gemma Mernihan and Brenton Cavanagh for help with confocal microscopy.
References (65)
- et al.
Budesonide loaded nanoparticles with pH-sensitive coating for improved mucosal targeting in mouse models of inflammatory bowel diseases
J. Control. Release
(2014) - et al.
Budesonide-loaded nanostructured lipid carriers reduce inflammation in murine DSS-induced colitis
Int. J. Pharm.
(2013) - et al.
Recent advances in inflammatory bowel disease therapy
Eur. J. Pharm. Sci.
(2020) - et al.
The burden of inflammatory bowel disease in Europe
J. Crohn's Colitis
(2013) - et al.
Nano-and microparticulate drug carriers for targeting of the inflamed intestinal mucosa
J. Control. Release
(2012) - et al.
Early mucosal healing with infliximab is associated with improved long-term clinical outcomes in ulcerative colitis
Gastroenterology
(2011) - et al.
Risk of surgery for inflammatory bowel diseases has decreased over time: a systematic review and meta-analysis of population-based studies
Gastroenterology
(2013) - et al.
Inflammatory bowel disease and targeted oral anti-TNFα therapy
Adv Protein Chem Struct Biol
(2020) - et al.
Acute effects of the cys-leukotriene-1 receptor antagonist, montelukast, on experimental colitis in rats
Eur. J. Pharmacol.
(2001) - et al.
PEG-functionalized microparticles selectively target inflamed mucosa in inflammatory bowel disease
Eur. J. Pharm. Biopharm.
(2013)
Site-specific targeted drug delivery systems for the treatment of inflammatory bowel disease
Biomed. Pharmacother.
Locally injected Infliximab ameliorates murine DSS colitis: differences in serum and intestinal levels of drug between healthy and colitic mice
Digest. Liver Dis.
Consumption of a baked corn and bean snack reduced chronic colitis inflammation in CD-1 mice via downregulation of IL-1 receptor, TLR, and TNF-α associated pathways
Food Res. Int.
Effect of surface chemistry on nanoparticle interaction with gastrointestinal mucus and distribution in the gastrointestinal tract following oral and rectal administration in the mouse
J. Control. Release
Alternating block copolymer-based nanoparticles as tools to modulate the loading of multiple chemotherapeutics and imaging probes
Acta Biomater
Comparative evaluation of novel biodegradable nanoparticles for the drug targeting to breast cancer cells
Eur. J. Pharm. Biopharm.
Gene silencing of TNF-alpha in a murine model of acute colitis using a modified cyclodextrin delivery system
J. Control. Release
Novel polyurethane-based nanoparticles of infliximab to reduce inflammation in an in-vitro intestinal epithelial barrier model
Int. J. Pharm.
Colectomy rate comparison after treatment of ulcerative colitis with placebo or infliximab
Gastroenterology
Nano-and microscaled particles for drug targeting to inflamed intestinal mucosa—A first in vivo study in human patients
J. Control. Release
Loss of prolyl hydroxylase-1 protects against colitis through reduced epithelial cell apoptosis and increased barrier function
Gastroenterology
Targeted delivery of the hydroxylase inhibitor DMOG provides enhanced efficacy with reduced systemic exposure in a murine model of colitis
J. Control. Release
Oral administration of pH-sensitive curcumin-loaded microparticles for ulcerative colitis therapy
Colloids Surf. B
Orally targeted delivery of tripeptide KPV via hyaluronic acid-functionalized nanoparticles efficiently alleviates ulcerative colitis
Mol. Ther.
Gastrointestinal stability of therapeutic anti-TNF α IgG1 monoclonal antibodies
Int. J. Pharm.
The global, regional, and national burden of inflammatory bowel disease in 195 countries and territories, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017
LancetGastroenterol. Hepatol.
Distinct cytokine patterns identified from multiplex profiles of murine DSS and TNBS-induced colitis
Inflamm Bowel Dis.
Novel Orally Administered Recombinant Anti-TNF Alpha Fusion Protein for the Treatment of Ulcerative Colitis: Results From a Phase 2a Clinical Trial
J. Clin. Gastroenterol.
imager: an R package for image processing based on CImg
J. Open Sour. Software
AVX-470: a novel oral anti-TNF antibody with therapeutic potential in inflammatory bowel disease
Inflamm. Bowel Dis.
Assessing mucosal inflammation in a DSS-induced colitis mouse model by MR colonography
Tomography
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