Sulfasalazine Research Articles

Protective antioxidative and anti-inflammatory actions of β-caryophyllene against sulfasalazine-induced nephrotoxicity in rat.

The pathogenesis of sulfasalazine (SFZ)-induced nephrotoxicity is unclear. Moreover, there are no reports on the protective effects of β-caryophyllene (BCP) against SFZ-induced renal injury. Hence, in this study, we measured several oxidative stress and inflammatory regulatory molecules alongside the effects of BCP in SFZ-intoxicated rats. Male rats (n = 48) were distributed to six equal groups as follows: negative control (NC), normal rats treated with low (N-LD; 200 mg/kg/day) and high (N-HD; 400 mg/kg/day) BCP doses, and animals treated with SFZ individually (PC; 600 mg/kg/day) or combined with BCP low (P-LD) and high (P-HD) doses. All drugs were administrated for 14 consecutive days. The NC, N-LD, and N-HD groups showed comparable renal histology and biochemistry. In contrast, abnormal histology, and increased creatinine and urea alongside oliguria and proteinuria were detected in the PC group. Renal specimens from the PC group revealed increased levels of nuclear factor-kappa B (NF-κB), transforming growth factor (TGF)-β with kidney injury molecule (KIM)-1, while the levels of nuclear factor erythroid 2-related factor 2 (Nrf2), AMP-activated protein kinase (AMPK), and protein kinase B (AKT) declined, relative to controls. The PC renal tissue also had markedly higher levels of inflammatory cytokines (tumor necrosis factor [TNF]-α/interleukin [IL]-1β/IL-6) and pro-oxidants (malondialdehyde [MDA]/H2O2/protein carbonyls), whereas those of antioxidants (glutathione [GSH]/glutathione peroxidase [GPx]/superoxide dismutase-1 [SOD1]/catalase [CAT]) and IL-10 decreased and were associated with marked apoptosis. Both BCP regimens ameliorated renal functions and histology, and reduced NF-κB, TGF-β, and KIM-1 levels in addition to those of oxidative stress and inflammation markers. Both protocols also augmented Nrf2, AMPK, AKT, antioxidants, and IL-10. However, P-HD showed better alleviating effects than the N-HD group. In conclusion, this study is the first to link NF-κB, TGF-β, Nrf2, AMPK, and AKT with SFZ-induced nephrotoxicity. In addition, this is the first report to reveal antioxidative and anti-inflammatory effects for BCP against SFZ-associated nephropathy.

Elaeocarpus sylvestris extract (ESE) inhibits inflammatory mediators and increases the efficacy of sulfasalazine in rheumatoid arthritis

BackgroundInterest in plant-based therapeutic products for treatment of several autoimmune diseases, including rheumatoid arthritis (RA), is increasing as current RA medications and treatments are either very expensive or ineffective. PurposeWe investigated the anti-inflammatory activity of Elaeocarpus sylvestris extract (ESE) and its therapeutic potential in collagen-induced arthritis (CIA) in the DBA/1 J mouse model and an in vitro model. MethodsIn the in vitro assay, the effects of ESE and a combination of ESE + sulfasalazine (SLZN) on nitric oxide and proinflammatory cytokine expression induced by lipopolysaccharide in RAW264.7 cells were confirmed. Furthermore, enzyme-linked immunosorbent assays and real-time polymerase chain reactions were used to measure levels of RA-related inflammatory cytokines and biomarkers in serum and knee joints to evaluate the changes caused by co-administration of ESE and SLZN in our mouse CIA model. Micro-CT scans of knee joints were analyzed for all groups. ResultsESE and the combination of ESE + SLZN downregulated the expression of lipopolysaccharide-induced nitric oxide and proinflammatory cytokines in RAW264.7 cells. Also, combined ESE and SLZN treatment improved arthritis score, edema volume, and proinflammatory cytokine serum levels compared with the CIA group. In addition, chemokine expression was downregulated compared with the CIA group. ConclusionThese findings suggest that coadministration of ESE and SLZN can improve the efficacy of arthritis treatment with reduced side effects.

Therapeutic Effects of Momordica charantia L. Ethanolic Extract on Acetic Acid-Induced Ulcerative Colitis in Rats

Objective: This study aims to investigate the effect of Momordica charantia L. (MoC) ethanolic extract on ulcerative colitis (UC) and was explored in vitro and in vivo. Materials and Methods: The rats were divided into control (C), saline-treated colitis (AA), MoC extract-treated colitis (AA+MoC), and sulfasalazine (SS)-treated colitis (AA+SS) groups. Colitis was induced by acetic acid. MoC extract, SS or saline were given to the related groups for 3 days. Interleukine-1β, malondialdehyde, glutathione levels, myeloperoxidase activity, bax/bcl-2 ratio, caspase-9 and caspase-3 levels were measured in colon and macroscopic and histopathologic examinations were done. Total phenolic/flavonoid content and biological activity of MoC were evaluated by in vitro analysis. Results: Compared to the control group, with acetic acid application interleukin-1β levels, myeloperoxidase activity, malondialdehyde levels, bax/bcl-2 ratio, caspase-9 and caspase-3 levels were significantly upregulated, while glutathione levels were significantly decreased in the AA group. In contrast, MoC and SS treatments reduced interleukin-1β, malondialdehyde levels, myeloperoxidase activity, bax/bcl-2 ratio, and caspase-9 and caspase-3 levels. Glutathione levels increased upon MoC or SS treatment. Increased macroscopic and microscopic scoring with AA improved with MoC or SS treatment, but the MoC or SS treated groups had higher score values than the control. Also, in vitro results showed that MoC exhibited 2,2-diphenyl-1- picrylhydrazyl and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging activity as well as significant antilipoxygenase activity. In addition, MoC extract showed a potent anti-inflammatory activity compared to standard indomethacin. Conclusion: Our biochemical, in vitro and histopathologic analysis indicate that MoC is likely to prove beneficial in UC therapy.

Checkpoint Inhibition Reduces the Threshold for Drug-Specific T-Cell Priming and Increases the Incidence of Sulfasalazine Hypersensitivity.

An emerging clinical issue associated with immune-oncology agents is the collateral effects on the tolerability of concomitant medications. One report of this phenomenon was the increased incidence of hypersensitivity reactions observed in patients receiving concurrent immune checkpoint inhibitors (ICIs) and sulfasalazine (SLZ). Thus, the aim of this study was to characterize the T cells involved in the pathogenesis of such reactions, and recapitulate the effects of inhibitory checkpoint blockade on de-novo priming responses to compounds within in vitro platforms. A regulatory competent human dendritic cell/T-cell coculture assay was used to model the effects of ICIs on de novo nitroso sulfamethoxazole- and sulfapyridine (SP) (the sulfonamide component of SLZ) hydroxylamine-specific priming responses. The role of T cells in the pathogenesis of the observed reactions was explored in 3 patients through phenotypic characterization of SP/sulfapyridine hydroxylamine (SPHA)-responsive T-cell clones (TCC), and assessment of cross-reactivity and pathways of T-cell activation. Augmentation of the frequency of responding drug-specific T cells and intensity of the T-cell response was observed with PD-1/PD-L1 blockade. Monoclonal populations of SP- and SPHA-responsive T cells were isolated from all 3 patients. A core secretory effector molecule profile (IFN-γ, IL-13, granzyme B, and perforin) was identified for SP and SPHA-responsive TCC, which proceeded through Pi and hapten mechanisms, respectively. Data presented herein provides evidence that drug-responsive T cells are effectors of hypersensitivity reactions observed in oncology patients administered ICIs and SLZ. Perturbation of drug-specific T-cell priming is a plausible explanation for clinical observations of how an increased incidence of these adverse events is occurring.

Chemical hybridization of sulfasalazine and dihydroartemisinin promotes brain tumor cell death

Gliomas are primary brain tumors with still poor prognosis for the patients despite a combination of cytoreduction via surgery followed by a radio-chemotherapy. One strategy to find effective treatment is to combine two different compounds in one hybrid molecule via linker to add to or at best potentiate their impact on malignant cells. Here, we report on the effects of a newly synthesized hybrid of sulfasalazine (SAS) and dihydroartemisinin (DHA), called AC254. In previous studies, both SAS and DHA have already proved to have anti-tumor properties themselves and to have sensitizing respectively potentiating effects on other treatments against malignant tumors. We investigated the impact of individual drugs SAS and DHA, their 1:1 combination and a novel SAS-DHA hybrid compound (AC254) on rodent and human glioma cells. In our study SAS alone showed no or only a mild effect on glioma, whereas DHA led to a significant reduction of cell viability in a dose-dependent manner. Next we compared the efficacy of the hybrid AC254 to the combinational treatment of its parent compounds SAS and DHA. The hybrid was highly efficient in combating glioma cells compared to single treatment strategies regarding cell viability and cell death. Interestingly, AC254 showed a remarkable advantage over the combinational treatment with both parent compounds in most used concentrations. In addition to its reduction of tumor cell viability and induction of cell death, the hybrid AC254 displayed changes in cell cycle and reduction of cell migration. Taken together, these results demonstrate that clinically established compounds such as SAS and DHA can be potentiated in their anti-cancer effects by chemical hybridization. Thus, this concept provides the opportunity to devise new effective chemotherapeutic agents.

Protective effect of mirtazapine against acetic acid-induced ulcerative colitis in rats: Role of NLRP3 inflammasome pathway

AimsUlcerative colitis (UC) is an inflammatory bowel disease (IBD) that causes long-lasting inflammation on the innermost lining of the colon and rectum. Mirtazapine (MRT) is a well-known antidepressant that was proven to have anti-inflammatory activity; however, to date, its role has not been investigated in UC. The current study aimed to investigate the role and mechanism of MRT in UC. Main methodAcetic acid (AA) was used for UC induction, and sulfasalazine (SLZ) was used as a positive control. Rats were divided into five equal groups; as follows; normal control, AA, SLZ (received SLZ in a dose of 250 mg/kg for 14 days), MRT10 (received MRT in a dose of 10 mg/kg/day for 14 days), and MRT30 (received MRT in a dose of 30 mg/kg/day for 14 days) groups. Macroscopic and microscopic examinations together with oxidative stress parameters evaluation were done. NOD-like receptors-3 (NLRP3), caspase-1, TNF-α, and nuclear factor kappa B (NF-κB) expression together with interleukin (IL)-1β and IL-18 levels were examined. Key findingMRT, in a dose-dependent manner, prevented the macroscopic and microscopic colonic damage and corrected the oxidative stress induced by AA. Moreover, MRT decreased the colonic tissue NLRP3 inflammasome, caspase-1, NF-κB, TNF-α expressions, IL-1β, and IL-18 levels that were elevated in colonic tissue by the AA. SignificanceMRT has a dose-dependent protective effect against UC that was mediated mainly by its anti-inflammatory activity with modulation of NLRP3/caspase-1 inflammatory pathway.

Treatment of ankylosing spondylitis during pregnancy

Aims of the trial – to study the frequency of drug use in pregnant women with ankylosing spondylitis (AS), to determine the effect of discontinuation of drugs of various groups, as well as the dose of non-steroidal anti-inflammatory drugs (NSAIDs) used, on AS activity during gestation.Material and methods. 50 pregnancies in 49 patients with AS that met the modified New York criteria of 1984. The average age of the patients was 31.7±4.9 years, the duration of the disease was 134.4±85.8 months. The visits were conducted at 10–11, 20–21 and 31–32 weeks of gestation. The BASDAI in the month of conception and in the I, II and III trimesters of pregnancy was: 1.4 [0.6; 3.3], 2.3 [1.2; 4.4], 2.8 [1.4; 4.2] and 2.2 [1.6; 4.0], respectively. The total dose of NSAIDs was determined by the NSAID intake index (Dougados M., 2011).Results and discussion. NSAIDs. After inclusion in the study, the drug of choice was ibuprofen, which was canceled for all women not later than on 32 week of gestation. At the time of conception and in the first, second and third trim. NSAIDs were taken by 23 (46%), 20 (40%), 30 (60%) and 21 (43.8%) women, respectively. NSAIDs intake index in I trimester (5.8 [2.9; 11.8]) was lower than before pregnancy (28.6 [16.7; 50]) and in II (15.5 [4.7; 30.9]) and III trimesters (24.4 [9.5; 50]) (p<0.05). No relationship between the index of ibuprofen intake, as well as the fact of withdrawal of NSAIDs and AS activity throughout gestation was found. Sulfasalazine (SS) in correspondence with arthritis was taken 3 months before conception by 11 (22%) women, during pregnancy – by 6 (12%) women at a dose of 1.25±0.25 g. Withdrawal of SS was not associated with recurrence of arthritis. Glucocorticoids (GC) at a dose of 7.5±2.5 mg 3 months before pregnancy and in the I, II and III trimesters of pregnancy were taken by 1 (2%), 4 (8%), 8 (16%) and 10 (20.8%) women, of whom 1 patient had arthritis and 1 had inflammatory bowel disease. The rest of the patients received GC due to high AS activity due to axial manifestations and the unavailability of TNFα inhibitors. Against the background of taking GC, the AS activity did not decrease: BASDAI in the II and III trimesters was 5.5±0.6 and 5.8±1.3 (p>0.05). TNFα inhibitors: 3 months before pregnancy and in the trim. of pregnancy were taken by 11 (22%), 7 (14%), 6 (12%) and 2 (4.2%) patients. In those who canceled therapy (both independently and on the recommendation of a rheumatologist) on the eve of pregnancy, an increase in AS activity was noted; BASDAI in the month of conception and in I, II, III trimesters was: 2.7 [0.8; 3.5], 5.1 [3.1; 5.9], 5.5 [5; 6] and 6.7 [5.3; 7.3] (p<0.05) compared to the month of conception. Cancellation of TNF-α in the month of conception was a risk factor for high AS activity (BASDAI>4) in the II trimester (OR=30.4; 95% CI: 1.5–612.3; p=0.03) and in the III trimester (OR=32.7; 95% CI: 1.6–662.2; p=0.02).Conclusion. NSAIDs and GC in low doses do not reduce the activity of AS. Withdrawal of TNF-α inhibitors on the eve of pregnancy is a predictor of high AS activity. It is necessary to increase the knowledge of rheumatologists and patients about the therapeutic possibilities during pregnancy to avoid unjustified drug withdrawal.

Chronic Sulfasalazine Treatment in Mice Induces System xc - - Independent Adverse Effects.

Despite ample evidence for the therapeutic potential of inhibition of the cystine/glutamate antiporter system xc − in neurological disorders and in cancer, none of the proposed inhibitors is selective. In this context, a lot of research has been performed using the EMA- and FDA-approved drug sulfasalazine (SAS). Even though this molecule is already on the market for decades as an anti-inflammatory drug, serious side effects due to its use have been reported. Whereas for the treatment of the main indications, SAS needs to be cleaved in the intestine into the anti-inflammatory compound mesalazine, it needs to reach the systemic circulation in its intact form to allow inhibition of system xc −. The higher plasma levels of intact SAS (or its metabolites) might induce adverse effects, independent of its action on system xc −. Some of these effects have however been attributed to system xc − inhibition, calling into question the safety of targeting system xc −. In this study we chronically treated system xc − - deficient mice and their wildtype littermates with two different doses of SAS (160 mg/kg twice daily or 320 mg/kg once daily, i.p.) and studied some of the adverse effects that were previously reported. SAS had a negative impact on the survival rate, the body weight, the thermoregulation and/or stress reaction of mice of both genotypes, and thus independent of its inhibitory action on system xc −. While SAS decreased the total distance travelled in the open-field test the first time the mice encountered the test, it did not influence this parameter on the long-term and it did not induce other behavioral changes such as anxiety- or depressive-like behavior. Finally, no major histological abnormalities were observed in the spinal cord. To conclude, we were unable to identify any undesirable system xc −-dependent effect of chronic administration of SAS.

Polyphyllin Ⅲ-Induced Ferroptosis in MDA-MB-231 Triple-Negative Breast Cancer Cells can Be Protected Against by KLF4-Mediated Upregulation of xCT.

Ferroptosis, which is characterized by the accumulation of intracellular iron and subsequent lipid peroxidation, is a newly discovered form of regulated cell death and plays an important role in tumor suppression. Herein, we showed that Polyphyllin III, which is a major saponin extracted from Paris polyphylla rhizomes, exerted its proliferation-inhibitory effect on MDA-MB-231 triple-negative breast cancer cells mainly through ACSL4-mediated lipid peroxidation elevation and ferroptosis induction. ACSL4 deletion partly attenuated Polyphyllin III-induced ferroptosis. Polyphyllin III treatment also induced KLF4-mediated protective upregulation of xCT, which is the negative regulator of ferroptosis. Interestingly, combination with the xCT inhibitor sulfasalazine (SAS) or downregulation of KLF4 sensitized MDA-MB-231 cells to Polyphyllin III. Furthermore, in vivo xenograft models, SAS significantly sensitized MDA-MB-231 breast cancer cells to Polyphyllin III, likely by enhancing intracellular lipid peroxidation and ferroptosis. The results of this study collectively demonstrated that Polyphyllin III exerts its anticancer effect by inducing ferroptosis via ACSL4 in MDA-MB-231 breast cancer cells. More importantly, we observed for the first time that KLF4-mediated xCT upregulation serves as negative feedback during ferroptosis progression, which might contribute to drug resistance in cancer treatment.

Studying the quenching resulted from the formation of an association complex between olsalazine or sulfasalazine with acriflavine.

We report the detection and quantification of important ulcerative colitis drugs olsalazine (OLS) and sulfasalazine (SUL) by the spectrofluorometric method. The proposed method was optimized and validated by using the quenching effect on the acriflavine fluorescence. The method was applied on the detection and quantification of OLS and SUL under optimized conditions showing the calibration curves were linear (range: 1.0–10.0 µg ml−1), with correlation coefficients R2 of 0.9999 for both drugs. The limits of detection (LOD) and quantification (LOQ) were 53 and 104 ng ml−1 for the OLS and 160 and 315 ng ml−1 for the SUL. This method permitted the analysis of OLS and SUL in their pure and pharmaceutical forms. The proposed spectrofluorimetric method was also evaluated against ‘green’ criteria and all the experimental results make it an eco-friendly and safe method for the detection of OLS and SUL.

The inhibition of spermatic cystine/glutamate antiporter xCT (SLC7A11) influences the ability of cryopreserved stallion sperm to bind to heterologous zonae pellucidae

Sperm are redox-regulated cells, and deregulation of their redox status is considered to affect male fertility and to reduce their fertilizing ability following biotechnological procedures, such as cryopreservation. Cystine (CysS), after incorporation in sperm via SLC7A11 antiporter, has been demonstrated to increase intracellular GSH content, the most important non enzymatic antioxidant. This study was aimed at investigating the role of SLC7A11 antiporter on frozen-thawed stallion sperm ability to respond to in vitro capacitating environment after post-thaw incubation with CysS and/or Sulfasalazine (SS), a specific inhibitor of SLC7A11 antiporter. Viability, motility, immunolocalization of tyrosine phosphorylated proteins and the ability to bind to heterologous zonae pellucidae were evaluated. Thawed sperm from seven stallions (2 ejaculates/stallion) was washed and resuspended in Tyrodes media; each thawed ejaculate was divided in Control (CTR) and 3 samples supplemented with: 0.5 mM Cystine (CysS), 500 μM Sulfasalazine (SS) and 0.5 mM CysS + 500 μM SS (CysS + SS). After 1 h of incubation at 37 °C, samples were washed twice, resuspended in capacitating BWW medium and incubated at 38 °C under 5% CO2. After 30 and 60 min, sperm motility, viability and tyrosine phosphorylated protein immunolocalization, used as capacitation status index, were evaluated. After 30 min of capacitation, 4 × 105 sperm were co-incubated with denuded pig oocytes in capacitation medium for 30 min for the heterologous binding assay. None of the sperm parameters studied (motility, viability and tyrosine phosphorylation) showed any difference respective to control. The number of sperm bound per oocyte (mean ± SEM) tended to increase in CysS group (44.0 ± 12.3) respect CTR (40.8 ± 10.8) while decreased in SS group (32.4 ± 7.8) (p < 0.01). Moreover, CysS + SS group showed a lower binding rate (32.0 ± 10.0) compared to CysS (p < 0.001). Our results suggest that CysS supplementation of thawed stallion sperm can influence their ability to bind to heterologous zona pellucidae as the inhibition of CysS incorporation by SLC7A11 reduced the number of sperm bound per oocyte. This effect does not seem to be ascribed to a modification of sperm motility, membrane integrity and tyrosine phosphorylation.

Host-guest chemistry of the sulfasalazine-β-cyclodextrin inclusion complex

Sulfasalazine (SLZ) is an azo drug used for the treatment of rheumatoid arthritis and ulcerative colitis. Its limited water solubility results into low permeability and bioavailability leading to higher daily recommended doses and greater side effects. Host-guest complexation of drugs with β-cyclodextrin (β-CD) is a common strategy to improve oral bioavailability and drug delivery. Traditional studies of inclusion complexes between SLZ and β-CD have been merely limited to UV analysis. We present herein, several structural evidences describing the detailed molecular architecture and binding constant of the inclusion complex using 1H NMR spectroscopy, isothermal titration calorimetry, ATR-IR spectroscopy, thermogravimetric analysis, and molecular modelling. Both 1H NMR and ITC methods converged toward closed values of Ka 1175 and 1220 M−1, respectively for the SLZ-β-CD inclusion complex. The modelling experiments also helped supporting the relative orientation of the SLZ into β-CD. The analysis revealed that SLZ integrates to β-CD exclusively from the benzoic acid end group and not through the pyridine moiety which is otherwise precluded because of its tilted structure that formed steric hindrance with the β-CD group. In addition, immobilization of SLZ through its benzoic acid side on CM5 sensor chip using surface plasmon resonance spectroscopy completely impeded the inclusion complex formation with β-CD. This comprehensive study of SLZ inclusion complexation with β-CD allows envisaging alternative strategies to improve SLZ solubility, bioavailability, and permeability for therapeutic applications.

Quantification and Metabolite Identification of Sulfasalazine in Mouse Brain and Plasma Using Quadrupole-Time-of-Flight Mass Spectrometry

Sulfasalazine (SAS), an anti-inflammatory drug with potent cysteine/glutamate antiporter system xc-(SXC) inhibition has recently shown beneficial effects in brain-related diseases. Despite many reports related to central nervous system (CNS) effect of SAS, pharmacokinetics (PK) and metabolite identification studies in the brain for SAS were quite limited. The aim of this study was to investigate the pharmacokinetics and metabolite identification of SAS and their distributions in mouse brain. Using in vivo brain exposure studies (neuro PK), the PK parameters of SAS was calculated for plasma as well as brain following intravenous and oral administration at 10 mg/kg and 50 mg/kg in mouse, respectively. In addition, in vivo metabolite identification (MetID) studies of SAS in plasma and brain were also conducted. The concentration of SAS in brain was much lower than that in plasma and only 1.26% of SAS was detected in mouse brain when compared to the SAS concentration in plasma (brain to plasma ratio (%): 1.26). In the MetID study, sulfapyridine (SP), hydroxy-sulfapyridine (SP-OH), and N-acetyl sulfapyridine (Ac-SP) were identified in plasma, whereas only SP and Ac-SP were identified as significant metabolites in brain. As a conclusion, our results suggest that the metabolites of SAS such as SP and Ac-SP might be responsible for the pharmacological effect in brain, not the SAS itself.

6-(methylsulfinyl)hexyl isothiocyanate (6-MITC) from Wasabia japonica alleviates inflammatory bowel disease (IBD) by potential inhibition of glycogen synthase kinase 3 beta (GSK-3β).

Inflammatory bowel disease (IBD) describes a set of disorders involving alterations to gastrointestinal physiology and mucosal immunity. Unravelling its complex pathophysiology is important since many IBD patients are refractory to or suffer adverse side effects from current treatments. Isothiocyanates (ITCs), such as 6-(methylsulfinyl)hexyl ITC (6-MITC) in Wasabia japonica, have potential anti-inflammatory activity. We aimed to elucidate the pathways through which 6-MITC alleviates inflammation by examining its role in the nuclear factor-kappa B (NF-κB) pathway through inhibition of glycogen synthase kinase 3 beta (GSK-3β) using a chemically induced murine model of IBD, cell-based and in silico techniques. The effects of 6-MITC and two NF-κB inhibitors, sulfasalazine (SS), pyrrolidine dithiolcarbamate (PDTC) were investigated on a dextran sulfate sodium (DSS)-induced murine mouse model of acute and chronic colitis using macroscopic measurements and pro-inflammatory markers. The effect of 6-MITC on NF-κB induction was assessed using a murine macrophage cell line. Complexes of GSK-3β-6-MITC and GSK-3β-ATP were generated in silico to elucidate the mechanism of 6-MITC's direct inhibition of GSK-3β. Changes in pro-inflammatory markers, inducible nitric oxide synthase (iNOS) (increased) and interleukin-6 (IL-6) (decreased) demonstrated that iNOS regulation occurred at the translational level. Intraperitoneal (ip) injection of 6-MITC to the colitis-induced mice ameliorated weight loss whereas oral administration had negligible effect. Fecal blood and colon weight/length ratio parameters improved on treatment with 6-MITC and the other NF-κB inhibitors. Levels of NF-κB decreased upon addition of 6-MITC invitro while structural studies showed 6-MITC acts competitively to inhibit GSK-3β at the ATP binding site. In this study we demonstrated that 6-MITC inhibits NF-κB signaling via GSK-3β inhibition ameliorating fecal blood, colonic alterations and DSS-induced weight loss indirectly indicating reduced intestinal stress. Taken together these results suggest a role for 6-MITC in the treatment of IBD acting to alleviate inflammation through the GSK-3β/NF-κB pathway. Furthermore, the GSK-3β-6-MITC model can be utilized as a basis for development of novel therapeutics targeting GSK-3β for use in other disorders including cancer.

MYCN-Amplified Neuroblastoma Is Addicted to Iron and Vulnerable to Inhibition of the System Xc-/Glutathione Axis.

MYCN is amplified in 20% to 25% of neuroblastoma, and MYCN-amplified neuroblastoma contributes to a large percent of pediatric cancer-related deaths. Therapy improvements for this subtype of cancer are a high priority. Here we uncover a MYCN-dependent therapeutic vulnerability in neuroblastoma. Namely, amplified MYCN rewires the cell through expression of key receptors, ultimately enhancing iron influx through increased expression of the iron import transferrin receptor 1. Accumulating iron causes reactive oxygen species (ROS) production, and MYCN-amplified neuroblastomas show enhanced reliance on the system Xc- cystine/glutamate antiporter for ROS detoxification through increased transcription of this receptor. This dependence creates a marked vulnerability to targeting the system Xc-/glutathione (GSH) pathway with ferroptosis inducers. This reliance can be exploited through therapy with FDA-approved rheumatoid arthritis drugs sulfasalazine (SAS) and auranofin: in MYCN-amplified, patient-derived xenograft models, both therapies blocked growth and induced ferroptosis. SAS and auranofin activity was largely mitigated by the ferroptosis inhibitor ferrostatin-1, antioxidants like N-acetyl-L-cysteine, or by the iron scavenger deferoxamine (DFO). DFO reduced auranofin-induced ROS, further linking increased iron capture in MYCN-amplified neuroblastoma to a therapeutic vulnerability to ROS-inducing drugs. These data uncover an oncogene vulnerability to ferroptosis caused by increased iron accumulation and subsequent reliance on the system Xc-/GSH pathway. SIGNIFICANCE: This study shows how MYCN increases intracellular iron levels and subsequent GSH pathway activity and demonstrates the antitumor activity of FDA-approved SAS and auranofin in patient-derived xenograft models of MYCN-amplified neuroblastoma.

Canna x generalis L.H. Bailey rhizome extract ameliorates dextran sulfate sodium-induced colitis via modulating intestinal mucosal dysfunction, oxidative stress, inflammation, and TLR4/ NF-ҡB and NLRP3 inflammasome pathways

Ethnopharmacological relevanceGenus Canna is used in folk medicine as demulcent, diaphoretic, antipyretic, mild laxative and in gastrointestinal upsets therapy. Canna x generalis (CG) L.H. Bailey is traditionally used as anti-inflammatory, analgesic and antipyretic. Besides, CG is used in Ayurvedic medicines’ preparations and in the treatment of boils, wounds, and abscess. Nevertheless, its anti-inflammatory effects against ulcerative colitis (UC) are not yet investigated. AimThis study aimed to investigate the phytoconstituents of CG rhizome ethanol extract (CGE). Additionally, we aimed to comparatively evaluate its therapeutic effects and underlying mechanisms against the reference drug “sulphasalazine (SAS)” in dextran sodium sulfate (DSS)-induced UC in mice. Material and methodsMetabolic profiling of CG rhizomes was performed via UHPLC/qTOF-HRMS; the total phenolic, flavonoid and steroid contents were determined, and the main phytoconstituents were isolated and identified. Next, DSS-induced (4%) acute UC was established in C57BL/6 mice. DSS-induced mice were administered either CGE (100 and 200 mg/kg) or SAS (200 mg/kg) for 7 days. Body weight, colon length, disease activity index (DAI) and histopathological alterations in colon tissues were examined. Colon levels of oxidative stress (GSH, MDA, SOD and catalase) and pro-inflammatory [Myeloperoxidase (MPO), nitric oxide (NO), IL-1β, IL-12, TNF-α, and INF-γ] markers were colourimetrically determined. Serum levels of lipopolysaccharide (LPS) and relative mRNA expressions of occludin, TLR4 and ASC (Apoptosis-Associated Speck-Like Protein Containing CARD) using RT-PCR were measured. Protein levels of NLRP3 inflammasome and cleaved caspase-1 were determined by Western blot. Furthermore, immunohistochemical examinations of caspase-3, NF-ҡB and claudin-1 were performed. ResultsMajor identified constituents of CGE were flavonoids, phenolic acids, phytosterols, beside five isolated phytoconstituents (β-sitosterol, triacontanol fatty alcohol, β-sitosterol-3-O-β-glucoside, rosmarinic acid, 6-O-p-coumaroyl-β-D-fructofuranosyl α-D-glucopyranoside). The percentage of the phenolic, flavonoid and steroid contents in CGE were 20.55, 6.74 and 98.09 μg of gallic acid, quercetin and β-sitosterol equivalents/mg extract, respectively. In DSS-induced mice, CGE treatment ameliorated DAI, body weight loss and colon shortening. CGE attenuated the DSS-induced colonic histopathological alternations, inflammatory cell infiltration and histological scores. CGE elevated GSH, SOD and catalase levels, and suppressed MDA, pro-inflammatory mediators (MPO and NO) as well as cytokines levels in colonic tissues. Moreover, CGE downregulated LPS/TLR4 signaling, caspase-3 and NF-ҡB expressions. CGE treatment inhibited NLRP3 signaling pathway as indicated by the suppression of the protein expression of NLRP3 and cleaved caspase-1, and the ASC mRNA expression in colonic tissues. Additionally, CGE restored tight junction proteins’ (occludin and claudin-1) expressions. ConclusionOur findings provided evidence for the therapeutic potential of CGE against UC. CGE restored intestinal mucosal barrier's integrity, mitigated oxidative stress, inflammatory cascade, as well as NF-ҡB/TLR4 and NLRP3 pathways activation in colonic tissues. Notably, CGE in a dose of 200 mg/kg was more effective in ameliorating DSS-induced UC as compared to SAS at the same dose.

Long-Standing Ulcerative Colitis and Sulphasalazine Treatment Complicated by Adult Celiac Disease

The risk of celiac disease may be increased in patients with inflammatory bowel disease (IBD), however, the precise pathogenetic mechanisms involved remain controversial. In this report, an elderly man with long-standing and extensive ulcerative colitis was treated with daily salazopyrine for over 30 years. His medication was eventually discontinued although endoscopic surveillance studies showed healed colitis with minimal inflammatory change and no dysplasia. He subsequently volunteered as an IBD control in a celiac disease research study. Endomysial and tissue transglutaminase antibodies were unexpectedly positive and a small bowel biopsy showed changes of celiac disease that later responded to a gluten-free diet. Earlier historical duodenal and ileal biopsies had been normal suggesting that changes were new, developing after cessation of the salazopyrine. This unusual presentation “unmasking” celiac disease after cessation of long-standing salazopyrine for colitis raises the potential that the drug may have acted to suppress the inflammatory process in celiac disease and may offer another alternate and inexpensive therapeutic approach.

Preparation of Fe@Fe2O3/3D graphene composite cathode for electrochemical removal of sulfasalazine

In the present study, heterogeneous electro-Fenton (EF) process was applied to remove the sulfasalazine (SU) pharmaceutical from aqueous solutions. In the first part, 3D graphene loaded with Fe@Fe2O3 core-shell nanowires (Fe@Fe2O3/3D-GO) was used as a cathode electrode in the EF process. Graphene oxide (GO) was synthesized for the synthesis of 3D graphene nanocomposites using the improved Hummers’ method and subsequently 3D graphene synthesized by the hydrothermal method using glycine. Finally, Fe@Fe2O3/3D-GO composite was synthesized and its properties were assessed by Scanning electron microscopy, Atomic force microscopy, Brunauer–Emmett–Teller, Fourier-transform infrared spectroscopy and X-ray diffraction methods. Then, the cathode electrode was prepared using the resulting composite and its performance was evaluated using Cyclic Voltammetry analysis. In the final part of this work, the Fe@Fe2O3/3D-GO electrode was used as the cathode electrode in the heterogeneous EF process to remove SU from aqueous solutions. The effect of operating parameters such as applied current (mA), initial pH of solution, initial pharmaceutical concentration (mg L−1) and process time (min) on pharmaceutical removal efficiency under heterogeneous EF process was investigated by response surface methodology. The results showed that the optimum values for applied current, pH, initial pharmaceutical concentration and electrolysis time were respectively 300 mA, 7, 30 mg L−1 and 100 min, resulting 99.60% of SU removal. Finally, the intermediates of SU degradation were determined by Gas chromatography–mass spectrometry analysis and the amount of mineralization was determined by total organic carbon analysis. About 5.2% drop in the SU removal efficiency was observed within 8 operational runs.

Combination of magnetic targeting with synergistic inhibition of NF-κB and glutathione via micellar drug nanomedicine enhances its anti-tumor efficacy

Breast cancer is not only one of the most prevalent types of cancer, but also it is a prime cause of death in women aged between 20 and 59. Although chemotherapy is the most common therapy approach, multiple side effects can result from lack of specificity and the use of overdose as safe doses may not completely cure cancer. Therefore, we aimed in this study is to combine the merits of NF-κB inhibiting potential of celastrol (CST) with glutathione inhibitory effect of sulfasalazine (SFZ) which prevents CST inactivation and thus enhances its anti-tumor activity. Inspired by the CD44-mediated tumor targeting effect of the hydrophilic polysaccharide chondroitin sulphate (ChS), we chemically synthesized amphiphilic zein-ChS micelles. While the water insoluble SFZ was chemically coupled to zein, CST was physically entrapped within the hydrophobic zein/SFZ micellar core. Moreover, physical encapsulation of oleic acid-capped SPIONs in the hydrophobic core of micelles enabled both magnetic tumor targeting as well as MRI theranostic capacity. Combining magnetic targeting to with the active targeting effect of ChS resulted in enhanced cellular internalization of the micelles in MCF-7 cancer cells and hence higher cytotoxic effect against MCF-7 and MDA-MB-231 breast cancer cells. In the in vivo experiments, magnetically-targeted micelles (154.4 nm) succeeded in achieving the lowest percentage increase in the tumor volume in tumor bearing mice, the highest percentage of tumor necrosis associated with significant reduction in the levels of TNF-α, Ki-67, NF-κB, VEGF, COX-2 markers compared to non-magnetically targeted micelles-, free drug-treated and positive control groups. Collectively, the developed magnetically targeted micelles pave the way for design of cancer nano-theranostic drug combinations.

The ZnO-NiO nano-composite: A brief characterization, kinetic and thermodynamic study and study the Arrhenius model on the sulfasalazine photodegradation

The coupled ZnO–NiO nano-composite (NC) was prepared and identified by powder X-ray diffraction (PXRD), scanning electron microscope (SEM), UV–Vis diffuse reflection spectroscopy (DRS) and Fourier transformation infrared spectroscopy (FTIR) characterization techniques. The crystallite sizes of about 6.7, 36.7 and 29.5 nm were obtained for the NiO, ZnO, and ZnO/NiO samples by the Williamson-Hall equation, and about of 33.5, 33.8 and 39.8 nm by the Scherrer equation, respectively. The energy dispersive X-Ray analysis (EDX) showed a NiO:ZnO mole ratio of 3.1 for the composite. The band gap energies of 3.49, 3.19, and 3.02 eV were estimated for the NiO, ZnO, and ZnO–NiO samples by using the absorption edge wavelengths of 355, 388, and 410, respectively. The pHpzc values of 9.3, 8.8, and 7.6 were also estimated for the NiO, ZnO, and ZnO–NiO samples respectively. At initial steps of the work about 42%, 60%, and 87% of a 10 mg/L SZ solution were degraded by the ZnO and NiO NPs and the ZnO–NiO NC, respectively. The best photocatalytic activity was obtained at pH 6.5 (near the pHpzc: 7.6), 0.7 g/L of the catalyst, and 7 mg/L sulfasalazine (SZ). The rate constants of 0.0904 min−1 (t1/2 = 7.66 min) and 0.0915 min−1 (t1/2 = 7.57 min) were obtained for the SZ photodegradation and mineralization processes, respectively. The frequency factor of 2.131 × 104 min−1 and the activation energy of 15.87 kJ/mol were obtained for the process by the Arrhenius equation. A negative activation ΔS (−0.22 kJ/mol) with the positive ΔH and ΔG values obtained for the SZ photodegradation by the ZnO–NiO composite.