Targeting Tumor Necrosis Factor-alpha Research Articles

PET biodistribution study of subcutaneous and intravenous administration of adalimumab in an inflammatory bowel disease model

Monoclonal antibodies targeting tumor necrosis factor-alpha (antiTNF-α) are used for patients with immuno-mediated illness as inflammatory bowel disease (IBD). However, 20–40 % of IBD patients do not respond to these therapies, and increasing knowledge of biodistribution could optimize their use and consequently their effectiveness. The aim of this study is to compare the biodistribution of adalimumab after intravenous (IV) and subcutaneous (SC) administration using Positron Emission Tomography (PET) imaging in IBD animal models. IBD was induced in mice using oral dextran sulfate sodium (DSS) and each induced animal was individually confirmed using [18F]FDG PET/CT scans, weight monitoring and histopathological analysis of colon tissue samples. The SC and IV biodistribution pharmacokinetics profiles and in vivo biodistribution of adalimumab labeled with 89Zr were evaluated using a dedicated PET/CT scanner. Mean standardized uptake values (SUV) were estimated from the colon, liver, and blood over seven days. Blood analysis revealed faster elimination of adalimumab in IBD models compared to controls, and after IV compared to SC administration (SUV 168 h p.i. SC-IBD = 0.06 ± 0.02, SC-Control = 1.08 ± 0.11, IV-IBD = 0.02 ± 0.01, IV-Control = 0.26 ± 0.13). Furthermore, IBD models exhibited faster whole-body clearance than controls and an earlier and higher concentration peak of adalimumab in the colon after IV (SUV 6 h p.i. IBD-IV = 2.11 ± 0.18) compared to SC administration (SUV 24 h p.i. IBD-SC = 1.49 ± 0.27). Our findings demonstrate the significant influence of the administration route and the TNF-α expression (local and also systemic) on the amount of adalimumab reaching the colon over time.

Oral administration of ruxolitinib in psoriasis vulgaris: A case report of plaque psoriasis accompanied by myelofibrosis secondary to polisitemia vera successfully treated with oral ruxolitinib.

Psoriasis is a chronic inflammatory skin disease characterized by keratinocyte hyperproliferation and immune cell infiltration. Various therapies have been discovered for psoriasis, including topical treatments, phototherapy, conventional systemic agents such as methotrexate, retinoids and ciclosporine, as well as biologics. Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway inhibitors targeting Tumor Necrosis Factor alpha (TNF-α), interleukin (IL)-23 and IL-17 can be effective in psoriasis. Ruxolitinib is a US Food and Drug Administration-approved first-generation Janus kinase inhibitor for polycythemia vera, myelofibrosis, and acute graft-versus-host disease. Ruxolitinib cream has been investigated in various dermatologic diseases, including atopic dermatitis, vitiligo, psoriasis, and alopecia areata. However, there is limited data on the efficacy of oral ruxolitinib in patients with psoriasis vulgaris. Here, we report a patient diagnosed with myelofibrosis coexisting with psoriasis vulgaris, successfully treated with oral ruxolitinib.

Evaluating the Efficacy of Infliximab in Inflammatory Bowel Disease: A Systematic Review of the Literature.

The introduction of steroid therapy in 1955 markedly decreased the mortality rate of severe ulcerative colitis (UC) from 24% in the placebo group to 7%, and it is currently less than 1% in specialist centers. Despite this advancement, the response of severe UC to steroids has stagnated over the past 50 years, with a high rate of colectomy persisting for severe to moderately severe cases. Infliximab (IFX) (Remicade, CentocorInc., Malvern, PA, United States), an intravenously administered chimeric monoclonal immunoglobulin G1 antibody targeting tumor necrosis factor-alpha (TNF-α), has shown efficacy in numerous randomized controlled trials for treating moderate to severe and fistulizing Crohn's disease, particularly in patients unresponsive to conventional therapies. This led to its approval by the US Food and Drug Administration in 1998 for treating active and fistulizing Crohn's disease. Most clinical research on IFX has focused on Crohn's disease, which is characterized as a Th1-type condition driven by pro-inflammatory cytokines like TNF-α. Conversely, UC has traditionally been viewed as a Th2-type condition where TNF-α plays a lesser role. However, recent studies indicate that TNF-α might also contribute to the pathogenesis of UC. These findings highlight the necessity for larger randomized controlled trials to further investigate the benefits of therapies like IFX, with the ultimate goal of improving treatment outcomes and quality of life for patients with inflammatory bowel disease.

Pain in axial spondyloarthritis: role of the JAK/STAT pathway.

Axial spondyloarthritis (axSpA) is a chronic inflammatory disease that is characterized by new bone formation in the axial musculoskeletal system, with X-ray discriminating between radiographic and non-radiographic forms. Current therapeutic options include non-steroidal anti-inflammatory drugs in addition to biological disease-modifying anti-rheumatic drugs that specifically target tumor necrosis factor-alpha (TNFα) or interleukin (IL)-17. Pain is the most critical symptom for axSpA patients, significantly contributing to the burden of disease and impacting daily life. While the inflammatory process exerts a major role in determining pain in the early phases of the disease, the symptom may also result from mechanical and neuromuscular causes that require complex, multi-faceted pharmacologic and non-pharmacologic treatment, especially in the later phases. In clinical practice, pain often persists and does not respond further despite the absence of inflammatory disease activity. Cytokines involved in axSpA pathogenesis interact directly/indirectly with the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling cascade, a fundamental component in the origin and development of spondyloarthropathies. The JAK/STAT pathway also plays an important role in nociception, and new-generation JAK inhibitors have demonstrated rapid pain relief. We provide a comprehensive review of the different pain types observed in axSpA and the potential role of JAK/STAT signaling in this context, with specific focus on data from preclinical studies and data from clinical trials with JAK inhibitors.

In Silico Identification of a Potential TNF-Alpha Binder Using a Structural Similarity: A Potential Drug Repurposing Approach to the Management of Alzheimer's Disease.

Alzheimer's disease (AD) is a neurodegenerative disorder with no conclusive remedy. Yohimbine, found in Rauwolfia vomitoria, may reduce brain inflammation by targeting tumour necrosis factor-alpha (TNFα), implicated in AD pathogenesis. Metoserpate, a synthetic compound, may inhibit TNFα. The study is aimed at assessing the potential utility of repurposing metoserpate for TNFα inhibition to reduce neuronal damage and inflammation in AD. The development of safe and effective treatments for AD is crucial to address the growing burden of the disease, which is projected to double over the next two decades. Our study repurposed an FDA-approved drug as TNFα inhibitor for AD management using structural similarity studies, molecular docking, and molecular dynamics simulations. Yohimbine was used as a reference compound. Molecular docking used SeeSAR, and molecular dynamics simulation used GROMACS. Metoserpate was selected from 10 compounds similar to yohimbine based on pharmacokinetic properties and FDA approval status. Molecular docking and simulation studies showed a stable interaction between metoserpate and TNFα over 100 ns (100000 ps). This suggests a reliable and robust interaction between the protein and ligand, supporting the potential utility of repurposing metoserpate for TNFα inhibition in AD treatment. Our study has identified metoserpate, a previously FDA-approved antihypertensive agent, as a promising candidate for inhibiting TNFα in the management of AD.

Biologic Medications for the Treatment of Psoriasis - Main Groups and Dosing System

This review explores the evolving landscape of psoriasis treatment with a focus on the transformative impact of biologic drugs. Psoriasis, a prevalent and persistent skin condition characterized by red and scaly patches, historically relied on topical, phototherapeutic, and systemic treatments, each with limitations. The advent of biologics represents a significant advancement, offering targeted interventions by addressing specific immunologic mechanisms underlying the disease. Biologics are now considered the preferred systemic therapy for chronic moderate-to-severe plaque psoriasis, particularly when conventional treatments prove ineffective or present disadvantages. The review delineates the mechanisms of action for biologics targeting tumour necrosis factor-alpha (TNF-α), interleukin-17 (IL-17) and interleukin-23 (IL-23). Specific drugs under each category, including etanercept, infliximab, adalimumab, secukinumab, ustekinumab, and others, are detailed with recommended dosages. Biologics have demonstrated substantial effectiveness, with clinical trials and real-world studies showcasing significant improvements in disease severity and patient’s quality of life. Notably, these drugs exhibit rapid action, often yielding noticeable changes within weeks. While biologics have revolutionized psoriasis treatment, the review emphasizes the importance of judicious use due to potential side effects such as injection-site reactions and respiratory infections. Serious adverse events, including infections and autoimmune reactions, necessitate careful patient selection and monitoring for safety. In conclusion, biologics offer a precise and effective approach to psoriasis treatment, promising marked symptom improvement and enhanced quality of life. The review underscores the need for responsible utilization, considering patient-specific factors, and anticipates ongoing advancements in biologics for improved control over this chronic dermatitis.

In vitro anti-inflammatory, antidiabetic, antibacterial, and in silico studies of Ferruginan A isolated from Olea ferruginea Royle (Oleaceae)

ObjectiveTraditionally, Olea ferruginea Royle (Oleaceae) has been used as a painkiller and antidiabetic in various ailments. To provide a scientific background to this folklore the current study was designed to anti-inflammatory and antidiabetic effects of one of the isolated compound from this plant. MethodsFerruginan A was isolated from the ethyl acetate extract of Olea ferruginea bark. This isolated molecule was subjected to in-vitro anti-inflammatory and antidiabetic effects using HRBCs and glucose uptake tests. The compound was also tested for molecular docking and ADMET study. ResultsRegarding the anti-inflammatory effect, the tested compound demonstrated a 69.82 % inhibition at a concentration of 100 µg/mL, while the Ferruginan A (100 µl/mL) increased the uptake of glucose (3.79–71.86 %) in the yeast cell. Similarly, the zone of inhibition values of Ferruginan A (24.98 mm) against Escherichia coli were found to be comparable to standard (Imipenem: 31.09 mm). The mechanism of antidiabetic and anti-inflammatory effects was explored by using docking simulations performed on four molecular targets related to diabetes and inflammation. The results showed that the isolated compound may act as an antidiabetic agent by inhibiting the 5′ Adenosine monophosphate-activated protein kinase (AMPK). While it also showed inhibition of anti-inflammatory targets COX-1, COX-2, and Tumor necrosis factor alpha (TNF-α). The ADMET prediction study revealed that isolated compound possesses favorable ADMET profile. ConclusionIt was concluded that Ferruginan A might be a significant anti-inflammatory and antidiabetic molecule.

Photoaffinity labeling coupled with proteomics identify PDI-ADAM17 module is targeted by (−)-vinigrol to induce TNFR1 shedding and ameliorate rheumatoid arthritis in mice

Various biological agents have been developed to target tumor necrosis factor alpha (TNF-α) and its receptor TNFR1 for the rheumatoid arthritis (RA) treatment, whereas small molecules modulating such cytokine receptors are rarely reported in comparison to the biologicals. Here, by revealing the mechanism of action of vinigrol, a diterpenoid natural product, we show that inhibition of the protein disulfide isomerase (PDI, PDIA1) by small molecules activates A disintegrin and metalloprotease 17 (ADAM17) and then leads to the TNFR1 shedding on mouse and human cell membranes. This small-molecule-induced receptor shedding not only effectively blocks the inflammatory response caused by TNF-α in cells, but also reduces the arthritic score and joint damage in the collagen-induced arthritis mouse model. Our study indicates that targeting the PDI-ADAM17 signaling module to regulate the shedding of cytokine receptors by the chemical approach constitutes a promising strategy for alleviating RA.

Hidradenitis Suppurativa: A Literature Review Comparing Current Therapeutic Modalities.

Hidradenitis suppurativa (HS) is a disease with a poor prognosis, often misinterpreted as an infection, with the highest impact on the patient's quality of life among all the assessed dermatological diseases. The main aim of this study was to compare various therapeutic interventions that are currently available for the treatment of HS. The pathogenesis of HS is not well understood, but it is mostly multifactorial involving a number of factors like genetic factors, androgens, local immunity, microflora, smoking, and obesity. Despite limited evidence on their effectiveness, topical antibiotics and antiseptics are commonly employed. Due to the colonization of bacteria and the presence of biofilms in the sinus tracts formed by HS lesions, systemic antibiotics are commonly employed as the primary form of therapy. In females with HS who experience menstrual flares or display symptoms of polycystic ovary syndrome, hormonal agents are often considered to be a viable and effective therapeutic option. At present, the sole treatment approved by both the Food and Drug Administration and the European Medicines Agency for addressing moderate to severe HS is adalimumab, an antibody that targets tumor necrosis factor alpha. Many surgical procedures in the management of HS aim to address inflammation by eliminating the affected folliculo-pilosebaceous unit, sinus tracts, and associated debris to impede further progression and scarring. HS continues to pose a considerable treatment challenge, necessitating a comprehensive approach for patients. However, the available evidence for most of these treatments is limited, indicating the need for more extensive research to identify the most effective interventions for managing HS.

Targeting Tumor Necrosis Factor Alpha to Mitigate Lung Injury Induced by Mustard Vesicants and Radiation.

Pulmonary injury induced by mustard vesicants and radiation is characterized by DNA damage, oxidative stress, and inflammation. This is associated with increases in levels of inflammatory mediators, including tumor necrosis factor (TNF)α in the lung and upregulation of its receptor TNFR1. Dysregulated production of TNFα and TNFα signaling has been implicated in lung injury, oxidative and nitrosative stress, apoptosis, and necrosis, which contribute to tissue damage, chronic inflammation, airway hyperresponsiveness, and tissue remodeling. These findings suggest that targeting production of TNFα or TNFα activity may represent an efficacious approach to mitigating lung toxicity induced by both mustards and radiation. This review summarizes current knowledge on the role of TNFα in pathologies associated with exposure to mustard vesicants and radiation, with a focus on the therapeutic potential of TNFα-targeting agents in reducing acute injury and chronic disease pathogenesis.

Psoriasis - a review of recent progress, characteristics, diagnostic management

Introduction: Psoriasis is a common, chronic, systemic inflammatory disease with an immunological basis. It occurs in about 2% of people in Europe and North America. It is associated with an increased probability of obesity, diabetes, dyslipidemia, cardiovascular disease and inflammatory bowel disease. Objective: The purpose of this review is to analyze the latest information on the characteristics, diagnosis and treatment of patients with psoriasis. Materials and methods: The present study was based on available data collected in the PubMed database and published between 2001 and 2022. The study was conducted by reviewing keywords such as: "psoriasis," "diagnosis," "treatment," "genetics," "immunology," "angiogenesis," "keratinocytes," and "incidence." State of the knowledge: In the last decade, biologic drugs targeting tumor necrosis factor alpha, interleukin 23, interleukin 17 and interleukin 12 have been developed and approved for the treatment of psoriasis. These drugs have revolutionized the treatment and management of psoriasis. Summary: Over the past decade, there have been tremendous advances in knowledge about the pathogenesis of psoriasis. Discoveries on this background, have given rise to new biological therapies. Currently in clinical trials are substances such as, Janus kinase inhibitors and tyrosine kinase 2 inhibitors, which represent hope for the most affected patients. Unfortunately, our knowledge of the causes of psoriasis is still incomplete. This poses a challenge for further research and thus the creation of more therapeutic substances.

USE OF INFLIXIMAB AS THERAPEUTIC OPTION FOR STEROID-RESISTANT PARADOXICAL REACTION IN TB

SESSION TITLE: TB and TB-Involved Case PostersSESSION TYPE: Case Report PostersPRESENTED ON: 10/17/2022 12:15 pm - 01:15 pmINTRODUCTION: Paradoxical reaction (PR) is an immune-mediated disease exacerbation precipitated by initiation of antitubercular therapy (ATT). Unlike IRIS (immune reconstitution inflammatory syndrome), a hyperinflammatory response to HIV/AIDS treatment, PR is independent of antiretroviral therapy (ART). Standard guideline therapy for PR is limited, with glucocorticoids as a standard of care. We describe a case of steroid-resistant tuberculin PR that improved with experimental Infliximab therapy.CASE PRESENTATION: 42-year-old HIV negative female with a history of rheumatoid arthritis on methotrexate and ovarian cancer on chemotherapy, presented with dyspnea and left-sided weakness. Chest imaging demonstrated a diffuse miliary pattern (image 1). QuantiFERON testing was positive. Cerebrospinal fluid studies, bronchoalveolar lavage and sputum cultures detected Mycobacterium tuberculosis (TB). Cultures demonstrated no antimicrobial resistance. Magnetic Resonance Imaging demonstrated lesions favoring CNS tuberculomas (image 2).There was transient clinical improvement with expanded ATT (Rifampin, Isoniazid, Pyrazinamide, Levofloxacin, Linezolid, Dexamethasone) but the disease continued to worsen despite increasing dosage of the glucocorticoid. Emergent salvage therapy with Infliximab significantly improved the intracranial and respiratory disease burden (image 3) with near resolution of clinical findings.DISCUSSION: An overexaggerated cell-mediated immune response against killed or damaged mycobacterium results in PR [1]. Moreover, granulomatous host response to TB may sequester the organism from administered antitubercular treatment [2]. Manipulation of immunomodulation by targeting tumor necrosis factor alpha (TNF alpha) is a proposed potential therapeutic option for PR [2] as it can decrease inflammation and disrupt granulomatous formations.CONCLUSIONS: There are only handful case reports exploring alternatives for steroid resistant PR. TNF-a inhibition is a potential valid treatment option in such cases [2].Reference #1: Blackmore, T. K., Manning, L., Taylor, W. J., & Wallis, R. S. (2008). Therapeutic Use of Infliximab in Tuberculosis to Control Severe Paradoxical Reaction of the Brain and Lymph Nodes. Clinical Infectious Diseases, 47(10), e83–e85. https://doi.org/10.1086/592695Reference #2: Briner, M., Oberholzer, M., Chan, A., & Wagner, F. (2021). Potential disease trigger as a therapeutic option: infliximab for paradoxical reaction in tuberculosis of the central nervous system. BMJ Case Reports, 14(8), e235511. https://doi.org/10.1136/bcr-2020-235511DISCLOSURES: No relevant relationships by Nicholas KlimbergNo relevant relationships by Pallabi ShresthaNo relevant relationships by Sarah UpsonNo relevant relationships by Kenji Yoshino SESSION TITLE: TB and TB-Involved Case Posters SESSION TYPE: Case Report Posters PRESENTED ON: 10/17/2022 12:15 pm - 01:15 pm INTRODUCTION: Paradoxical reaction (PR) is an immune-mediated disease exacerbation precipitated by initiation of antitubercular therapy (ATT). Unlike IRIS (immune reconstitution inflammatory syndrome), a hyperinflammatory response to HIV/AIDS treatment, PR is independent of antiretroviral therapy (ART). Standard guideline therapy for PR is limited, with glucocorticoids as a standard of care. We describe a case of steroid-resistant tuberculin PR that improved with experimental Infliximab therapy. CASE PRESENTATION: 42-year-old HIV negative female with a history of rheumatoid arthritis on methotrexate and ovarian cancer on chemotherapy, presented with dyspnea and left-sided weakness. Chest imaging demonstrated a diffuse miliary pattern (image 1). QuantiFERON testing was positive. Cerebrospinal fluid studies, bronchoalveolar lavage and sputum cultures detected Mycobacterium tuberculosis (TB). Cultures demonstrated no antimicrobial resistance. Magnetic Resonance Imaging demonstrated lesions favoring CNS tuberculomas (image 2). There was transient clinical improvement with expanded ATT (Rifampin, Isoniazid, Pyrazinamide, Levofloxacin, Linezolid, Dexamethasone) but the disease continued to worsen despite increasing dosage of the glucocorticoid. Emergent salvage therapy with Infliximab significantly improved the intracranial and respiratory disease burden (image 3) with near resolution of clinical findings. DISCUSSION: An overexaggerated cell-mediated immune response against killed or damaged mycobacterium results in PR [1]. Moreover, granulomatous host response to TB may sequester the organism from administered antitubercular treatment [2]. Manipulation of immunomodulation by targeting tumor necrosis factor alpha (TNF alpha) is a proposed potential therapeutic option for PR [2] as it can decrease inflammation and disrupt granulomatous formations. CONCLUSIONS: There are only handful case reports exploring alternatives for steroid resistant PR. TNF-a inhibition is a potential valid treatment option in such cases [2]. Reference #1: Blackmore, T. K., Manning, L., Taylor, W. J., & Wallis, R. S. (2008). Therapeutic Use of Infliximab in Tuberculosis to Control Severe Paradoxical Reaction of the Brain and Lymph Nodes. Clinical Infectious Diseases, 47(10), e83–e85. https://doi.org/10.1086/592695 Reference #2: Briner, M., Oberholzer, M., Chan, A., & Wagner, F. (2021). Potential disease trigger as a therapeutic option: infliximab for paradoxical reaction in tuberculosis of the central nervous system. BMJ Case Reports, 14(8), e235511. https://doi.org/10.1136/bcr-2020-235511 DISCLOSURES: No relevant relationships by Nicholas Klimberg No relevant relationships by Pallabi Shrestha No relevant relationships by Sarah Upson No relevant relationships by Kenji Yoshino

The Structural, Biological, and In-Silico Profiling of Novel Capryloyl Tetra-Glucoside and Aliphatic Ester Constituents from the Abutilon indicum Offers New Perspectives on the Treatment of Pain and Inflammation

Abutilon indicum L. (Malvaceae), more often referred to as Peeli booti, Kanghi, and Kakhi, is a perennial shrub found in many countries of Asia. Traditionally, this plant is used as a diuretic to treat inflammation, discomfort, urethral infections, and gout. Inflammation and pain are key topics of interest for researchers throughout the globe, since they are linked to almost every illness that could affect humans or animals. The present study was conducted to isolate the phytoconstituents from the methanolic extract of Abutilon indicum collected from the Bihar state Koshi river belt in India, and to evaluate the isolated phytoconstituents’ ability to reduce nociception and inflammation. Furthermore, molecular docking was performed to investigate the molecular interaction profile, with possible therapeutic targets for anti-inflammatory medicines. A. indicum methanolic extract yielded two novel phytocompounds identified as 5′-hydroxyhexyl n-hexadecanoate (AB-01) and n-octanoyl-β-D-glucopyranosyl-(2′-1′′)-β-D-glucopyranosyl-(2′′-1′′′)-β-D-glucopyranosyl-(2′′′-1′′′′)-β-D-glucopyranoside (AB-05), together with three previously recognized phytocompounds such as ester glucoside. All isolated molecules were tested for the efficacy of analgesic and anti-inflammatory characteristics at doses of 5 and 10 mg/kg body weight. The isolated compound’s molecular interaction profile with anti-inflammatory drug targets cyclooxygenase-2 and tumor necrosis factor-alpha possessed high potential energy in molecular docking. These findings may aid in developing anti-inflammatory and analgesic drugs from A. indicum.

A Lysosome-Activated Tetrahedral Nanobox for Encapsulated siRNA Delivery.

Tetrahedral framework nucleic acids (tFNAs) have attracted extensive attention as drug nanocarriers because of their excellent cellular uptake. However, for oligonucleotide cargos, tFNA mainly acts as a static delivery platform generated via sticky-ended ligation. Here, inspired by the original stable space inside the tetrahedral scaffold, a dynamic lysosome-activated tFNA nanobox is fabricated for completely encapsulating a short interfering RNA (siRNA) of interest. The closed tetrahedral structure endows cargo siRNA with greater resistance against RNase and serum and enables solid integration with the vehicle during delivery. Moreover, the pH-responsive switch of nanobox allows the controlled release of siRNA upon entry into lysosomes at cell culture temperature. Based on protective loading and active unloading, an excellent silencing effect on the target tumor necrosis factor alpha (TNFα) gene is achieved in in vitro and in vivo experiments. Conclusively, the nanobox offers a dynamic pH-sensitive confinement delivery system for siRNA and can be an extendable strategy for other small RNA.

Assessment of immunogenicity and drug activity in patient sera by flow-induced dispersion analysis

Biopharmaceuticals have revolutionized the treatment of many diseases such as diabetes, cancer, and autoimmune disorders. These complex entities provide unique advantages like high specificity towards their target. Unfortunately, biopharmaceuticals are also prone to elicit undesired immunogenic responses (immunogenicity), compromising treatment efficacy as well as patient safety due to severe adverse effects including life threatening conditions. Current immunogenicity assays are hampered by immobilization procedures, complicated sample pre-treatment, or rely on cell-based methods which all prevent reliable and continuous monitoring of patients. In this work, we present Flow Induced Dispersion Analysis (FIDA) for assessment of immunogenicity and drug activity in serum samples from arthritis patients receiving adalimumab. FIDA is a first principle technique for size-based characterization of biomolecules and their complexes under biologically relevant conditions. The FIDA methodology rely on an absolute and quantitative readout (hydrodynamic radius) thus reducing the need for positive and negative controls. Here, FIDA is applied for evaluating active adalimumab in serum by studying the interaction with its target tumor necrosis factor alpha (TNF-α). We report proof of principle for a quantitative approach for stratifying patients exhibiting presence of neutralizing and non-neutralizing antibodies based on their individual drug activity pattern. Further, it can be applied to any biopharmaceutical having soluble drug targets and it holds potential in a companion diagnostics setting.

P543 Point-of-care testing in therapeutic drug monitoring of infliximab

Abstract Background Within the past 20 years infliximab (IFX), a monoclonal antibody targeting tumor necrosis factor-alpha, has become an established therapy for inducing and maintaining remission in inflammatory bowel disease (IBD). IFX therapeutic drug monitoring (TDM) using the point-of-care (POC) methods showed to be able to produce results fast enough to allow IFX dose adjustments before drug infusion. This study aimed to compare IFX quantification using two POC tests, Procise IFX (Procise Dx) and Quantum Blue Infliximab assay (Bühlmann Laboratories AG). Methods Serum samples from 22 IBD patients (M=64%) on IFX maintenance therapy were collected immediately before drug infusion. Capillary blood was also collected from 6 IBD patients by finger prick using whole blood pipettes (Procise Dx). Crohn’s disease (CD) was diagnosed in 12 patients and ulcerative colitis (UC) in 10 patients. For method comparison, a Passing Bablok regression was used and for qualitative comparison weighted kappa statistic was obtained after stratification of results by therapeutic range (<3 mg/l, ≥3 to 7 mg/L, and ≥7 mg/L). Results Both methods measured lower IFX concentrations in CD patients (4,95 (0,65 – 5,80) mg/L) than in UC patients (8,50 (4,80 – 12,20) mg/L). In two patients with confirmed IFX antibodies, both methods showed immeasurably low IFX concentrations. Passing Bablok regression analysis of IFX concentration in serum sample has showed a proportional deviation between two POC methods (y=0.416 (-0.654 to 1.279) + 0.753 (0.648 to 0.923)x). A good comparison has been observed for capillary blood measured using Procise IFX test and serum samples measured using Quantum Blue Infliximab assay (y=-2.732 (-4.315 to 1,867) + 1.263 (0.278 to 1.566)x), although IFX concentration was lower in capillary blood. Classification of results according to therapeutic interval showed good agreement for serum samples and serum and whole capillary blood measurements (κ=0.79 (0.59–0.999 and κ=0.80 (0.45–1.00), respectively). Conclusion Because of proportional differences, two POC tests can’t be used interchangeably for longitudinal TDM of IFX in IBD patients. There is a need for caution in the interpretation of IFX in whole blood, due to its possibly lower concentration.

Molecular Docking Analysis of Ficus religiosa Active Compound with Anti-Inflammatory Activity by Targeting Tumour Necrosis Factor Alpha and Vascular Endothelial Growth Factor Receptor in Diabetic Wound Healing

BACKGROUND: Diabetes mellitus contributes to the delayed healing of wounds causes disturbance of inflammatory cytokine. Tumour necrosis factor alpha (TNF-alpha) and Vascular Endothelial Growth Factor Receptor (VEGFR) both have a role in the persistent inflammation associated with diabetic wounds. Ficus religiosa has developed a reputation as a traditional wound healer among some java people in Indonesia. AIM: Our study aims to discover the molecular interaction between the active constituents of F. religiosa with TNF-alpha and VEGFR. MATERIALS AND METHODS: This research was conducted in computerized molecular docking using Protein database, Pymol, Discovery studio, and Pyrex software. A thorough literature search was conducted to identify the potential compound and molecular target for diabetic wounds. Analysis of its anti-inflammatory properties was also carried out using a passonline webserver. Pharmacokinetic analysis was performed using the Lipinski Rule of Five websites and the PreADMET website. RESULTS: Each of the study’s active compounds has a good pharmacokinetic profile. The predictions of the compound’s structure indicate that it has a strong anti-inflammatory impact. Lupenyl acetate and Lanosterol bind more strongly to the TNF-alpha than the natural ligand, but Piperine binds more strongly to VEGFR. CONCLUSIONS: Lupenyl acetate, Lanosterol, and Piperine compounds have anti-inflammatory effects through inhibition of TNF-alpha and VEGFR. In addition, this compound has potential to become a drug because it has good pharmacokinetics. Future studies are required to determine the effectiveness and toxicity of Lupenyl acetate, Lanosterol, and Piperine as potential treatment in diabetic wounds.

A Review of the Totality of Evidence for the Development and Approval of ABP710 (AVSOLA), an Infliximab Biosimilar.

ABP 710 (AVSOLA®) is a biosimilar to infliximab reference product (RP), a monoclonal antibody targeting tumor necrosis factor alpha (TNFα). It is approved in the USA and Canada for all the same indications as infliximab RP. Approval of ABP 710 was based on the totality of evidence (TOE) generated using a stepwise approach to assess its similarity with infliximab RP with regard to analytical (structural and functional) characteristics, pharmacokinetic parameters, and clinical efficacy and safety. ABP 710 was shown to be analytically similar to infliximab RP including in amino acid sequence, primary peptide structure, and glycan mapping and purity. ABP 710 was also demonstrated to be similar to infliximab RP with regard to functional characterization including in vitro binding, effector functions, and signaling pathways important for the mechanisms of action for clinical efficacy in multiple indications of immune-mediated inflammatory disorders including inflammatory bowel disease (IBD), especially binding to both soluble and membrane-bound TNFα. Pharmacokinetic similarity of ABP 710 with infliximab RP was demonstrated in healthy volunteers following a single 5 mg/kg intravenous dose. Comparative clinical efficacy of ABP 710 with infliximab RP was demonstrated in patients with rheumatoid arthritis. Safety and immunogenicity were also demonstrated to be similar for both ABP 710 and the RP. Overall, the TOE supported the conclusion that ABP 710 is highly similar to infliximab RP and supported scientific justification for extrapolation to all approved indications of infliximab RP, including IBD.

Альтернативні методи у веденні прееклампсії. Аналітичний огляд

There are many alternative drugs for the prevention and treatment of preeclampsia in the new research. This drugs can effect on the underlying pathophysiology of the disease: oxidative stress, antiangiogenic factors, as well as angiotensin, nitric oxide and various parts of the inflammatory process. Thus, they affect the disease of the placenta or endothelium. The proposed treatments are currently undergoing preclinical and clinical trials. Pravastatin was of the greatest interest among all the proposed therapeutic agents. It has pleiotropic effect, i.e. affects multiple molecular targets against preeclampsia. Proton pump inhibitors, metformin, and sulfasalazine are other drugs that have preclinical evidence of multiple molecular actions that may address the pathophysiology of preeclampsia. Currently, these molecules are also in clinical trials. Many natural compounds for the treatment of preeclampsia, such as plant extracts and trace elements, are being researched to identify the potential in anti-inflammatory or antioxidant activity. Monoclonal antibodies are another direction is new molecular-oriented strategies. They are targeting tumor necrosis factor alpha, placental growth factor and short interfering RNA technology to inhibit the expression of soluble fms-like tyrosine kinase-1 or angiotensinogen. Folic acid, nitric oxide donors (such as L-arginine), recombinant antithrombin III, and immunogenic digoxin antigen and melatonin are other treatment approaches that have been tested in humans (ranging from single-group studies to phase III trials that have been completed or are ongoing). The series of cases demonstrated that removal of circulating soluble fms-like tyrosine kinase-1 can help stabilize the disease and prolong pregnancy. Monoclonal antibodies such as eculizumab (a complement inhibitor) may have therapeutic potential. Thus, the identified alternative drugs in the treatment and prevention of preeclampsia create the potential to improve maternal health and pregnancy. No conflict of interests was declared by the author. Key words: preeclampsia, pregnancy, pravastatin, metformin, sulfalazine

Fucosylation and Sialylation of Fc-Fragment of anti-Tumour Necrosis Factor Alpha Antibodies do not Influence Their Immunogenicity in Monocyte-Derived Dendritic Cells.

Antibodies targeting tumor necrosis factor-alpha [TNF-alpha] are a mainstay in the treatment of inflammatory bowel disease. However, they fail to demonstrate efficacy in a considerable proportion of patients. On the other hand, glycosylation of antibodies might influence not only their immunogenicity but also their structure and function. We investigated whether specific glycosylation patterns of the Fc-fragment would affect the immunogenicity of anti-TNF-alpha antibody in monocyte-derived dendritic cells. The effect of a specific Fc-glycosylation pattern on antibody uptake by monocyte-derived dendritic cells [mo-DCs] and how this process shapes the immunologic profile of mo-DCs was investigated. Three N-glycoforms of the anti-TNF-alpha antibody adalimumab, that differed in the content of fucose or sialic acid, were tested: [1] mock treated Humira, abbreviated 'Fuc-G0', where the N-glycan mainly consist of fucose and N-acetylglucosamine [GlcNAc], without sialic acid; [2] 'Fuc-G2S1/G2S2' with fucose and alpha 2,6 linked sialic acid; and [3] 'G2S1/G2S2' with alpha 2,6 linked sialic acid, without fucose. Our data demonstrated that neither fucosylation nor sialylation of anti-TNF-Abs [Fuc-G0, FucG2S1/G2S2, G2S1/G2S2] influence their uptake by mo-DCs. Additionally, none of the differentially glycosylated antibodies altered CD80, CD86, CD273, CD274 levels on mo-DCs stimulated in with lipopolysaccharide in the presence of antibodies. Next, we evaluated the levels of cytokines in the supernatant of mo-DCs stimulated with lipopolysaccharide in the presence of Fuc-G0, Fuc-G2S1/G2S2 or G2S1/G2S2-glycosylated anti-TNF antibodies. Only IL-2 and IL-17 levels were downregulated, and IL-5 production was upregulated by uptake of Fuc-G0 antibodies, as compared to control without antibodies. The specific modification in the Fc-glycosylation pattern of anti-TNF-alpha Abs does not affect their immunogenicity under the tested conditions. As this study was limited to mo-DCs, further investigation is required to clarify whether Ab uptake into mo-DCs might change the immunological profile of T- and B-cells, in order to ultimately reduce the formation of anti-drug antibodies and to improve the patient care.