Targeting the ileo-colonic region in inflammatory bowel disease

Inflammatory bowel diseases (IBD) includes Crohn’s disease (CD) and ulcerative colitis (UC). IBD are chronic inflammatory diseases of the gastrointestinal tract (GIT). The inflammation in IBD is generally localized in the GIT. Therefore, topical therapy with anti-inflammatory drugs is the desired therapeutic approach in many cases. However, depending on disease type (UC or CD) and severity, the inflammation may be present throughout the entire GIT, such as the small intestine, ileum, and/or colon. This means that for an effective topical treatment, the drug should be targeted to a given site in the GIT. Earlier studies have shown that the ColoPulse coating technology targets oral dosage forms, such as tablet and capsules, to the ileo-colonic region of healthy subjects and CD patients. In this thesis we used this coating to develop three novel tablets that contain topically active anti-inflammatory drugs in view of maximizing the local anti-inflammatory effects in the GIT, whilst minimizing the systemic effects that may result in adverse events. The tablets contained a combination of mesalazine and budesonide (formulation 1), or budesonide only (formulation 2), or the monoclonal antibody infliximab (formulation 3). This thesis investigated the drug release characteristics, ileo-colonic targeting performance, and stability of these new formulations. The results showed that these novel formulations are interesting new treatment options for IBD that may be more efficacious than the currently available budesonide and mesalazine formulations since none are optimally suited to treat the ileo-colonic region. Ileo-colonic targeting of infliximab has not yet been investigated. However, animal studies as well as small pilot studies suggest that this may be an efficacious treatment option that eliminates the unwanted adverse events that are related to systemic exposure. In a review article we summarize all the available data that support this notion. However, this needs to be investigated in a clinical trial. The study protocol of such a trial is part of this thesis

Equipment

This chapter is about the design, quality and application of equipment for the preparation of medicines in a pharmacy or for preparation in small scale pharmaceutical industry. The type of pharmaceutical equipment needed depends on the type of products to be produced, on the required productive capacity and the batch size. A list of essential and critical equipment for production and quality control must be included as attachment in the URS (User Requirements Specification) of any facility. The equipment, requirements, qualification methods, main applications, maintenance and cleaning procedures are described for: • Powder exhaust units, Laminar airflow units, Safety cabinets and Isolators • Pharmaceutical water production • Storage and distribution of pharmaceutical water • Heating and Ultrasonic water baths • Grinding, mixing and dispersing • Filling, dosing and closing for liquids, suppositories, capsules and tubes • Fridges and freezers • Automatic filling and robotics • 3D printing.</p

The Development and Implementation of Airflow Visualization Studies ("Smoke" Studies) as a Training Tool in Aseptic Hospital Compounding Facilities

In the compounding facilities of hospital pharmacies, extemporaneous preparations for parenteral administration are produced using aseptic handling. The designated environment for this practice is a clean area, such as a laminar airflow (LAF) cabinet placed in a classified cleanroom complying with good manufacturing practices (GMP) and International Organization for Standardization (ISO) 14644-1 guidelines. The European GMP Annex 1 (Revision 2020) and United States Pharmacopeia (USP) monograph state that airflow visualization studies (“smoke” studies) should be performed to substantiate the cleanroom and LAF cabinet performance and their qualification status. Even though smoke studies are required by these guidelines, current literature does not describe detailed practical protocols and acceptance criteria. The objective of this study was to develop and implement a practical smoke study protocol to ensure compliance with aseptic handling guidelines in hospital pharmacies. First, a literature search was performed to collect information about smoke study protocols and acceptance criteria. Subsequently, a smoke study protocol was developed for a downflow and crossflow LAF cabinet as well as for grade C/B cleanroom areas. As a proof of concept, the smoke study protocol for the downflow LAF cabinet was executed in the at-rest and in-operation states. Video recordings of the smoke studies were analyzed to assess the performance of the cabinet. Finally, the video recordings obtained from the smoke studies were used in a training program for hospital pharmacy operators, which showed that smoke studies might aid in operators’ aseptic handling awareness. To the best of our knowledge, the present study provides for the first time a practical approach for the development of smoke study protocols in a hospital pharmacy setting and shows potential for training operators, process optimization, and continuous quality improvement

Review:Local Tumor Necrosis Factor-alpha Inhibition in Inflammatory Bowel Disease

Crohn’s disease (CD) and ulcerative colitis (UC) are inflammatory bowel diseases (IBD) characterized by intestinal inflammation. Increased intestinal levels of the proinflammatory cytokine tumor necrosis factor-α (TNF-α) are associated with disease activity and severity. Anti- TNF-α therapy is administered systemically and efficacious in the treatment of IBD. However, systemic exposure is associated with adverse events that may impede therapeutic treatment. Clinical studies show that the efficacy correlates with immunological effects localized in the gastrointestinal tract (GIT) as opposed to systemic effects. These data suggest that site-specific TNF-α inhibition in IBD may be efficacious with fewer expected side effects related to systemic exposure. We therefore reviewed the available literature that investigated the efficacy or feasibility of local TNF-α inhibition in IBD. A literature search was performed on PubMed with given search terms and strategy. Of 8739 hits, 48 citations were included in this review. These studies ranged from animal studies to randomized placebo-controlled clinical trials. In these studies, local anti-TNF-α therapy was achieved with antibodies, antisense oligonucleotides (ASO), small interfering RNA (siRNA), microRNA (miRNA) and genetically modified organisms. This narrative review summarizes and discusses these approaches in view of the clinical relevance of local TNF-α inhibition in IBD

Infliximab formulation strategy for a stable ileo-colonic targeted oral dosage form intended for the topical treatment of inflammatory bowel disease

Research shows that topically active infliximab therapy may be efficacious in the treatment of inflammatory bowel disease (IBD) with expected fewer side effects related to systemic exposure. Oral administration of infliximab with site-specific delivery could enable such therapy. In the present study, infliximab was incorporated in a sugar glass matrix (IFX-I) for additional stability and compounding flexibility after which IFX-I was compounded to ileo-colonic-targeted tablets containing 5 mg infliximab (ColoPulse-IFX). Potential critical steps in the production process that may decrease the formulation stability were identified and investigated. Furthermore, the long-term stability of IFX-I (6 months) and ColoPulse-IFX (12 months) stored either at room temperature (25 degrees C +/-+/- 2 degrees C/60% RH +/- 5% RH) or refrigerated (5 degrees C +/- 3 degrees C) was investigated according to ICH guidelines. Size-exclusion chromatography, fluorescence spectroscopy, and ELISA analyses were used to investigate the infliximab stability, content, tertiary protein structure, and potency at t0, t3, t6, t9, and t12 months. The coating performance of ColoPulse-IFX was investigated in a gastrointestinal simulation system at t0 and t12 months. All the analyses showed that IFX-I and ColoPulse-IFX were stable, potent, and that the coating performance was maintained during the entire storage period at both storage conditions. Thus, IFX-I is a stable drypowder formulation and ColoPulse-IFX is a promising oral dosage form for the topical treatment of ileocolonic IBD. This formulation strategy may serve as a new platform for the development of oral peptide or protein formulations that are targeted to the ileo-colonic region in IBD.</p

Validation of an LC-MS/MS assay for rapid and simultaneous quantification of 21 kinase inhibitors in human plasma and serum for therapeutic drug monitoring

Kinase inhibitors have revolutionized cancer treatment in the past 25 years and currently form the cornerstone of many treatments. Due to the increasing evidence for therapeutic drug monitoring (TDM) of kinase inhibitors, the need is growing for new assays to rapidly evaluate kinase inhibitor plasma concentrations. In this study, we developed an LC-MS/MS assay for the rapid and simultaneous quantification of 21 kinase inhibitors. First, a literature search was conducted to ensure that the linear ranges of the analytes were in line with the reported therapeutic windows and/or TDM reference values. Subsequently, the assay was validated according to FDA and EMA guidelines for linearity, selectivity, carry-over, accuracy, precision, dilution integrity, matrix effect, recovery, and stability. The assay was fast, with a short run-time of 2 min per sample. Sample pre-treatment consisted of protein precipitation with methanol enriched with stable isotope-labeled internal standards (SIL-IS), and the mixture was vortexed and centrifuged before sample injection. Separation was achieved using a C18 column (3 μm,50 × 2.1 mm) with a gradient of two mobile phases (ammonium formate buffer pH 3.5 and acetonitrile). Analyte detection was conducted in positive ionization mode using selected reaction monitoring. The assay was accurate and precise in plasma as well as in serum. Extraction recovery ranged between 95.0% and 106.0%, and the matrix effect was 95.7%-105.2%. The stability of the analytes varied at room temperature and in refrigerated conditions. However, all drugs were found to be stable for 7 days in the autosampler. The clinical applicability of the analytical method (486 analyzed samples between 1 July 2022-1 July 2023) as well as external quality control testing results were evaluated. Taken together, the results demonstrate that the analytical method was validated and applicable for routine analyses in clinical practice.</p

Development of a Personalized Tumor Neoantigen Based Vaccine Formulation (FRAME-001) for Use in a Phase II Trial for the Treatment of Advanced Non-Small Cell Lung Cancer

Stage III-IV non-small cell lung cancer (NSCLC) is a devastating disease characterized by a poor prognosis. NSCLC tumors carry genetic mutations, which can lead to the expression of altered protein sequences. Peptides originating from mutated proteins and bound to MHC molecules on the tumor cell surface are referred to as neoantigens, as they are tumor-specific and not expressed in normal cells. Due to their tumor specificity, neoantigens have a strong potential to induce an anti-tumor immune response and have been investigated for development of personalized therapeutic cancer vaccines. The current study describes the development of a clinical grade neoantigen vaccine formulation (FRAME-001) intended as immunotherapy in advanced NSCLC in combination with the immune checkpoint inhibitor pembrolizumab. The detection of aberrant tumor-specific transcripts as well as an algorithm to select immunogenic neoantigen peptides are described. Subsequently, selected neoantigen peptides were synthesized with a high throughput synthesis platform and aseptically formulated under good manufacturing practice (GMP) conditions into four aqueous peptides mixtures that each contained six neoantigen peptides. A validated stability-indicating analytical method was developed in which we considered the personalized nature of the formulation. An extensive stability study performed either at -25 degrees C or -80 degrees C showed that the formulation was stable for up to 32 weeks. The formulation was mixed with the vaccine adjuvant Montanide ISA 51 VG, which yielded the final vaccine emulsion. The stability of the vaccine emulsion was demonstrated using microscopic examination, differential light scattering, and the water-drop test. The presented data show that FRAME-001 is a feasible personalized vaccine formulation for the treatment of stage III-IV NSCLC. The presented data may give guidance in the development of novel personalized therapeutic vaccines since this formulation strategy could be used for any cancer indication