Recent Advances in Inflammation & Allergy Drug Discovery - Volume 15, Issue 2, 2021

Advancements in stem cell research have enabled the establishment of three-dimensional (3D) primary cell cultures, known as organoids. These culture systems follow the organization of an in vivo organ, as they enclose the different epithelial cell lines of which it is normally composed. The generation of these 3D cultures has bridged the gap between in vitro models, made up of two-dimensional (2D) cancer cell lines cultures, and in vivo animal models, with major differences regarding human diseases. Organoids are increasingly used as a model to study colonization of gastric mucosa by infectious agents and better understand host-microbe interactions and the molecular events that lead to infection, pathogen-epithelial cells interactions, and mechanisms of gastric mucosal injury. In this review, we will focus on the role of organoids as a tool to investigate molecular interactions of Helicobacter (H.) pylori and Epstein Barr Virus (EBV) and gastric mucosa and how these infections, which affect ≈ 45 % of the world population, might progress to gastric cancer, a highly prevalent cancer and the third leading cause of cancer death.

Background: Inflammation plays a significant role in the pathophysiology of Acute Coronary Syndrome (ACS) but is not included in current risk stratification. Objective: This study aimed at determining the association between Thrombolysis in Myocardial Infarction (TIMI) risk score and inflammatory biomarkers in the ACS, including unstable angina (UA), Non-ST Segment Elevation Myocardial Infarction (NSTEMI), and ST-segment elevation myocardial infarction (STEMI). We hypothesized that inflammatory biomarkers could add prognostic value to the TIMI risk score. Methods: In this cross-sectional study, serum levels of interleukins (IL)-6 and IL-1β and MDA (malondialdehyde) were quantified by ELISA and colorimetry, respectively, of patients with ACS (n = 48; 31.3 % with UA, 33.3 % with NSTEMI, and 35.4 % with STEMI) and healthy controls (n = 43). We assessed the TIMI scores in the first 24 h after symptom onset. Results: The results showed that patients with ACS had significantly higher levels (p<0.05) of the inflammatory biomarkers IL-6, IL-1β, and MDA than the control group. However, we found no significant differences in IL-6, IL-1β, and MDA levels among the patients with ACS according to their classification as UA, NSTEMI, and STEMI. Positive correlations were observed between TIMI and IL-6 (r=0.68), IL-1β (r= 0.53), and MDA (r=0.58) in patients with UA and between TIMI and IL-1β (r= 0.62) in STEMI patients. Conclusion: These data suggested the presence of a pro-inflammatory profile in patients with ACS as well as positive correlations between TIMI scores and the inflammatory biomarkers IL-6, IL-1β, and MDA in patients with UA and between TIMI scores and IL-1β in patients with STEMI. Combining inflammatory biomarkers with the TIMI risk score could provide better insight into the processes involved in ACS.

Background: The organism’s defense against aggressive agents is performed by the innate immune system via activation of pattern-recognition receptors (PRRs). Initially, these agents are recognized by the immune system, resulting in the inflammatory response that activates the pathogen elimination and tissue repair. Inflammasomes are macromolecules related to the host’s response to endo or exogenous aggressive agents. Thus, inflammation mediated by inflammasomes plays an important role in the pathogenesis of diseases, such as neurodegenerative disorders, autoimmune diseases, and type 2 diabetes, justifying their attractiveness as drug targets. One of the most important tasks remains in the ATPase nucleotide-binding oligomerization domain nucleotide- binding domain leucine-rich repeat-containing receptors protein 3 (NLRP3), in which the blocking of its oligomerization is related to the functional inhibition of inflammasomes. Here, we performed molecular docking and dynamics simulations for NP3-146, an analog of MCC950, to obtain information about the complex stability and main interactions with amino acid residues from NLRP3. Methods: By using the crystalized structure recently deposited in the protein data bank (7alv), molecular docking in GOLD software and molecular dynamics simulations in GROMACS software were performed to generate the RMSD RMSF, Rg, SASA, and H-bond plots. Results: The results of RMSD, RMSF, Rg, SASA, and H-bond plots of both complexes confirmed the stability at the active site. Besides, the analyses of the most stable conformation showed that the main interactions are performed with Ala²²⁷, Ala²²⁸, Pro³⁵², Ile⁴¹¹, Phe⁵⁷⁵, and Arg⁵⁷⁸ residues. Conclusion: This report confirmed the stability of NP3-146, similar to the known inhibitor MCC950, providing useful information for designing NLRP3 inhibitors.

Aims: To prepare solid lipid nanopaticles (SLNs) of Ketoprofen (KP) using microwave method. Ketoprofen (KP) is 2-(3-benzolphenyl) propionic acid with anti-inflammatory, analgesic and antipyretic property. The drug has a short half-life of 120 mins. It belongs to BCS Class II drug. Gastric irritation is a major limitation for delivery because of acidic nature of the drug. The development of solid lipid nanoparticles with its transdermal drug delivery was the aim of the present work. Methods: Microwave-assisted microemulsion technique was used for the development of solid lipid nanoparticles. Stearic acid was used as lipid and tween 80 was used as a surfactant. By varying the type of lipid and input energy watt, batches were formulated. SLNs were evaluated for zeta potential, drug entrapment, particle size and in-vitro drug release. Crystallinity behaviour was determined by differential scanning calorimetry and powder X-ray diffraction. Anti-inflammatory activity was evaluated for batch M4 of SLNs. The gel was prepared for M4 batch. It was evaluated for viscosity, pH, drug content, in-vitro and ex-vivo diffusion study. Results: SLNs were developed successfully. Based on the size, entrapment efficiency, stability and drug release, batch M4 was selected. SLNs showed 74.8% entrapment efficiency. Forty-fold improvement was observed in the solubility. The particle size was 682.9 nm and average size 1047 nm. PDI was 0.685 and zeta potential was -29.5 mV. M4 SLNs batch of gel showed burst release followed by a controlled release for 8 hrs in in-vitro drug release. Conclusion: SLNs were successfully prepared by Microwave-assisted microemulsion technique. SLNs with anti-inflammatory activity were successfully developed with their transdermal delivery.

Background: Inflammation is the response to the reaction of any type of bodily injury by elevating cellular metabolism and releasing soluble mediators. It is also a contributing factor of pain. Predimenol, which has previously been known as DLBS1442, is a bioactive extract from Phaleria macrocarpa (Scheff.) Boerl (Thymelaceae). It can be an alternative treatment for pain relief, especially for long-term use. Objective: The objective of this study is to evaluate the anti-inflammatory activities of predimenol through the evaluation of several parameters involved in the inflammatory pathway. Methods: Cyclooxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor- α (TNF-α), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-6 (IL-6), and nuclear factor ΚB (NF-ΚB) were observed after 24 h exposure of predimenol (0-180 μg/mL) to lipopolysaccharides (LPS)-activated RAW 264.7 cell. The inflammatory markers were measured using nitric oxide (NO) assay and enzyme-linked immunosorbent assay (ELISA) for COX-2 inhibitor assay. The gene expressions of TNF-α, IL-1β, IL-2 and IL-6 were quantified using the polymerase chain reaction (PCR) method. Western blotting was applied to detect phosphorylated IΚB kinase (IKK) protein to confirm the activation of NF-ΚB. Results: Our study showed a similar mechanism with most non-steroidal anti-inflammatory drugs (NSAIDs). Predimenol consistently downregulated the expression of iNOS and inhibited COX-2 activity. Moreover, predimenol significantly inhibited the LPS-induced production of NO, TNF-α, IL-1β, IL-2 and IL-6. Down-regulation of these markers was suggested due to the reduction of NF- ΚB transcription level and activation by predimenol. Conclusion: Predimenol exhibits anti-inflammatory activities through NF-ΚB inactivation-mediated COX-2 suppression, which may suggest that predimenol is a potential analgesic and anti-inflammatory agent.

Objective: The main objectives of the present work are to determine the clinical effect of niranthin on visceral or somatic inflammatory pain. The study was performed to determine the effects of niranthin on visceral or somatic inflammatory hypersensitivity of adult Swiss albino mice by using complete Freund’s adjuvant (CFA) induced pain model. Methods: The effect of CFA injection was determined after 24 hours of injection by using an aesthesiometer such as Von Frey filaments to evaluate tactile acetone-evoked cooling and thermal sensitivity. We used a digital Plethysmometer to measure paw edema. Single dose of niranthin intraperitoneal injection (5 and 10 mg/kg) was injected into mice having CFA-induced mechanical hypersensitivity and after 30 minutes of administration, reduced mechanical hypersensitivity was observed. In addition, niranthin also reduced acetone-evoked hypersensitivity within 4 hours. Results: Compared to DMSO, niranthin was most highly active to reduce CFA-induced paw edema. To reduce mechanical hypersensitivity, multiple doses of niranthin (bis in die (b.i.d.)) from 1st - 5th day and b.i.d. day 9th and 10th) were given and remarkable results were observed such as did not cause tolerance in multiple dosing and significantly reduced in CFA induced hypersensitivity. Conclusion: This work reported niranthin having antinociceptive activity and indicated that niranthin is conventionally active in the management of persistent pain.