Research in Pharmaceutical Sciences最新文献

Background and purpose: Actinobacteria are highly valuable in the pharmaceutical industry due to their unlimited capacity to produce natural products. In line with the screening of actinomycetes to discover new antibacterial agents, this study isolated and elucidated bioactive compounds from two Streptomyces strains, F9 (99.88% similarity with S. chryseus) and F4 (99.14% similarity with S. rectiviolaceus).

Experimental approach: The present study followed a rigorous experimental approach. Previously, two Streptomyces strains, F9 and F4, were isolated and identified. These strains were then selected for the isolation and elucidation of secondary metabolites. The crude extract was semi-purified using Waters, and the active fractions were further purified using Agilent. The structure of the isolated compounds was elucidated through detailed spectroscopic analysis to ensure the accuracy and reliability of the findings.

Findings/results: Four fractions isolated from strain F9 showed antibacterial activity against test microorganisms. HR-ESI-MS, fragmentation pattern, and database search identified chrysomycins B and C. The structure of chrysomycins A and 4'-O-acetylated A was confirmed by 1D and 2D-NMR data. In addition, the findings characterized 2 pigments produced by strain F4 using HR-ESI-MS and fragmentation pattern (ESI-MS/MS), which revealed that major and minor peaks corresponded to butyl-meta-cycloheptylprodiginine and undecylprodiginine, respectively.

Conclusion and implications: Six bioactive compounds, including 4 chrysomycins and 2 pigments, were isolated from 2 Streptomyces strains, F9 and F4. Importantly, this was the first report on isolating chrysomycins A-C and 4'-O-acetylated A from S. chryseus.

Background and purpose: The N-terminal Hsp90 inhibitors are promising targets for cancer treatment; however, inducing the heat shock response is one of the most significant limitations. A prominent way to overcome this limitation is by inhibiting the Hsp90 C-terminal domain.

Theoretical approach: In this study, a set of structure-based methods was engaged to predict the new C-terminal inhibitors. Since there was no human PDB structure of the Hsp90 C-terminal domain, homology modeling was done using the SWISS-MODEL server online. The 3D structure of the model was refined through energy minimization using molecular dynamics (MD) simulation for 10 ns. The active site of the created model was validated by novobiocin docking. Four steps of virtual screening, including HTVS, SP, XP, and QM/MM docking, were performed on the created library (151,332 compounds) based on 80% similarity to deguelin as the C-terminal inhibitor. The best-obtained compounds were introduced to MM-GBSA studies. Finally, the stability of the best compound was investigated using a 100 ns MD simulation.

Results/findings: Four steps of virtual screening were performed on the created library. The extracted 46 compounds with the XP GlideScore of < -4.164 kcal/mol were introduced to MM-GBSA studies, and rescoring was done. The stability of compound CID_14018348, the best compound (ΔG_binding = -80.45 kcal/mol), was investigated using MD simulation.

Conclusion and implications: The compound CID_14018348 was identified as the most promising candidate through computational techniques; therefore, the computational methods outlined can be applied in the development of potent anticancer agents.

Background and purpose: Inflammatory bowel disease (IBD) is characterized by aberrant immune responses in the colon, leading to the inflammatory cascades. Metformin (MTF) and crocin demonstrated beneficial effects in reducing inflammation and oxidative stress, as the main triggers of IBD. Considering the coloprotective properties of MTF and crocin alone, the present study aims to investigate the possible therapeutic effects of combination therapy with MTF and crocin on the acetic acid-induced colitis model.

Experimental approach: Acute colitis was induced in Wistar rats by intrarectal administration of acetic acid. Nine study groups were assessed, including normal group received normal saline, control group received normal saline after colitis induction, dexamethasone as reference (1 mg/kg), MTF-treated groups received 100, 150, 200 mg/kg of drug, crocin-treated received 20 and 30 mg/kg of crocin, and combination therapy received MTF (150 mg/kg) + crocin (20 mg/kg). Colon tissues were collected to assess macroscopic, microscopic, and biochemical parameters.

Findings/results: Our data revealed that the combination therapy, crocin-treated and MTF-treated (at the higher dose) groups, ameliorated disease severity by decreasing myeloperoxidase activity and malondialdehyde level, compared to the control group. Combination therapy was also effective in attenuating macroscopic parameters, including ulcer index as well as wet weight of the colon. Histopathological scores considerably decreased in all groups.

Conclusion and implications: MTF, crocin, and their combined administration exerted ameliorative effects in the colons of acetic acid-induced colitis rats, which is probably due to their anti-inflammatory and antioxidant properties. Further experimental studies are required to elucidate the underlying mechanisms.

Background and purpose: Multiple sclerosis (MS) is a degenerative disease of the central nervous system. Fatigue is one of the most common and disabling symptoms of MS. Garlic is a plant whose anti-fatigue effects have been shown. The present study aimed to investigate the potential effectiveness of garlic on fatigue and quality of life in MS patients.

Experimental approach: In a randomized, single-blind, placebo-controlled clinical trial, adult MS patients were randomly divided into two groups: of drug (garlic) group and a placebo. The drug group received 400-mg tablets of garlic extract (equivalent to 1,200 μg of allicin) twice a day for 4 weeks, while the patients in the placebo group received placebo tablets at the same frequency and duration. Before and after the intervention, scores on a 36-item survey form (SF-36) and fatigue severity scale (FSS) questionnaires were recorded for all patients and compared between the groups.

Findings/results: Garlic consumption was significantly associated with an increase in energy/fatigue, pain, general health, and physical health subscales scores at the end of the intervention compared to the placebo group. The scores of FSS were significantly reduced in both groups; however, the change in the drug group was remarkably higher than in the placebo group.

Conclusion and implications: Garlic extract promotes fatigue and improves the quality of life in MS patients. Therefore, garlic can be considered a potential remedy to overcome fatigue and improve the quality of life in these patients.

Background and purpose: Cigarette smoking induces lung toxicity by triggering oxidative stress, leading to apoptosis, ferroptosis, and senescence. Sulforaphane (SFN), a potent antioxidant, activates the SIRT1 pathway, enhancing cellular stress resistance and survival. This study aimed to evaluate the protective effects of SFN against cigarette smoke extract (CSE)-induced damage in human airway epithelial cells (BEAS-2B) and in mouse lungs, focusing on its role in upregulating SIRT1 expression.

Experimental approach: BEAS-2B cells were treated with CSE and SFN, and cell viability was assessed using the MTT assay. Cellular senescence was assessed using the SA-β-gal assay and the expression of genes associated with senescence (p16 and p21). The expression levels of SIRT1, senescence-associated secretory phenotype (SASP) cytokines (IL-1β, IL-6, IL-8, TNF-α), GPX4, and SLC7A11 were quantified using qRT-PCR. Additionally, ROS production, GSH and MDA levels, and iron content were measured. An emphysema mouse model was induced by intraperitoneal administration of CSE (7.2 mg/kg) alone or in combination with SFN (10.2 mg/kg) over 28 days, and subsequent histopathological changes were evaluated.

Findings/results: Our findings revealed that SFN co-treatment effectively mitigated CSE-induced cytotoxicity, senescence, and SASP cytokine secretion, as well as the pronounced emphysematous changes in lung tissues. Furthermore, SFN reversed CSE-induced downregulation of SIRT1 and upregulation of NF-κB. Notably, SFN also inhibited CSE-induced ferroptosis by increasing GPX4 and SLC7A11 expression while reducing iron and MDA levels.

Conclusion and implications: The findings of the present study demonstrated that sulforaphane offers protective effects against CSE-induced toxicity by mitigating oxidative stress, ferroptosis, and cellular senescence.

Background and purpose: Dill (Anethum graveolens) is an herbal plant from the Apiaceae family often used as an effective remedy for several ailments. This study aimed to investigate the potential protective effect of dill against the development of non-alcoholic fatty liver disease in obese rats.

Experimental approach: For 12 weeks, rats were fed a high-fat diet (HFD) to induce obesity. In the treatment group, the extract of dill leaves (100 mg/kg) was administered by gavage. Then, blood and liver samples were harvested for further investigations.

Findings/results: Feeding HFD caused increased body mass index, abdominal circumference, adiposity index, weight gain, serum glucose, lipids, insulin, leptin, and insulin resistance. HFD-fed rats also showed increased hepatic triglycerides, fatty acid synthase, cytochrome P2E1, hydrogen peroxide, malondialdehyde, serum marker enzymes (AST, ALT, ALP, and GGT), and liver weight, with decreased antioxidants including superoxide dismutase, catalase, and glutathione. Besides, a significant elevation of hepatic interleukins 1β and 6,tumor necrosis factor-α, nuclear factor-kappa B, Kupfer cell markers (CD68 and CD163), fibronectin, andcollagen type 1, along with an increase of transforming growth factor-β1 expression, was observed. Histological changes presented by hepatocytes, including ballooning, inflammatory cell aggregation, and deposition of collagen fibers, have also been detected. Co-administration of dill with HFD succeeded in reducing weight gain, hepatic triglyceride accumulation, oxidative reactions, inflammation, fibrosis, and liver structural injury.

Conclusion and implications: Dill extract could be approved as a promising therapeutic approach with multiple benefits for the management of obesity and associated steatohepatitis.

Background and purpose: Peroxisome proliferator-activated receptors alpha and gamma (PPAR-α and PPAR-γ) are nuclear receptor proteins that play a crucial role in the regulation of cellular differentiation, development, metabolism, and tumorigenesis. Their expression levels have been implicated in the metabolic reprogramming of breast cancer cells, influencing their proliferation and survival. This study investigates the expression of PPAR-α and PPAR-γ in breast cancer and explores their relationship with key enzymes involved in fatty acid biosynthesis: fatty acid synthase (FASN), acyl-CoA synthetase long-chain family member (ACSL4), and ATP citrate lyase (ACLY).

Experimental approach: In this study, 28 pairs of fresh samples of breast cancer and adjacent non-cancerous tissue were analyzed to assess gene expression levels using quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry staining.

Findings/results: The expression of PPAR-α increased, while PPAR-γ decreased significantly in breast cancer tissues compared to adjacent normal tissues. The expression of PPAR-α was significantly associated with FASN mRNA expression. Additionally, a correlation was also observed between the expression levels of both PPAR-α and PPAR-γ with ACSL4 mRNA levels.

Conclusion and implications: Given the obtained results, the involvement of PPARs in the regulation of lipid metabolism was substantiated. Moreover, the correlation of PPARs with ACSL4 highlights the possible role of PPAR-α and PPAR-γ in the regulation of tumor tissue ferroptosis and suggests that targeting these pathways could offer new therapeutic strategies for managing breast cancer. However, further studies are needed to understand the mechanism of action.

Background and purpose: Pluronic F127-based hydrogel is a fair formulation for increasing the protein stability and half-life without decreasing its biological activity. The present study aimed to prepare FP127 hydrogel loaded with the recombinant BIF1-iRGD protein as a new immunotoxin with targeting potential of cancer cells.

Experimental approach: BIF1-iRGD in 19% w/v of FP127 was prepared by the cold method, and its in vitro release was determined. MTT and flow-cytometry assays were performed to evaluate the cytotoxic and apoptotic effects of BIF1-iRGD and BIFl-iRGD-hydrogel against 4T1 cells. The tumor size of 4T1 Balb-C mice was evaluated, and H&E staining was used for histopathology evaluation.

Findings/results: BIFl-iRGD release followed the first-order model. The toxicity of BIFl-iRGD was less than that in the formulation as a hydrogel after 48 and 72 h of treatment. The null hydrogel showed no toxicity compared to the untreated cells. After 24 h, cells treated with BIFl-iRGD-hydrogel and BIFl-iRGD around their 48-h IC₅₀ value developed apoptosis at about 55% and 35%, respectively. The tumor size decreased over time in the treated mice during the 20 days after the last injection. Also, no significant difference was observed between the effectiveness of the two groups. Tumor sections of mice treated with BIF1-iRGD and BIF1-iRGD-hydrogel had necrotic parts of about 65% and 70%, respectively.

Conclusion and implications: BIF-iRGD-hydrogel showed in vivo anticancer effects with increased toxicity in comparison to the native protein. However, the investigation of protein distribution and probable cytotoxic effect on vital organs must be noted.

Background and purpose: Antimicrobial resistance poses a significant global health threat. A previously developed penta-amino acid ultrashort antimicrobial peptide (UP5), with alternating arginine and biphenylalanine units, showed strong antimicrobial activity, particularly in combination with levofloxacin. However, differences in pharmacokinetics between UP5 and levofloxacin may hinder their optimal clinical use. This study aimed to develop a covalent UP5-levofloxacin conjugate that retains the synergistic antimicrobial properties of both UP5 and levofloxacin.

Experimental approach: The UP5-levofloxacin conjugate was synthesized and tested for antimicrobial activity against multidrug-resistant gram-positive (Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis) and gram-negative bacteria (Acinetobacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa). Hemolytic activity was assessed on human erythrocytes, and selectivity against MDCK cells was determined. Comparative analysis was performed with individual components.

Findings/results: The conjugate exhibited synergistic antimicrobial activity with minimum inhibitory concentration (MIC) values between 2.5 and 20 μM, overcoming levofloxacin resistance. It showed minimal hemolytic activity < 1% and a favorable selectivity index of 4.4-35.2. Cytotoxicity studies indicated selective toxicity towards MDCK cells, with an IC₅₀ of 88.34 μM, significantly higher than its MIC values.

Conclusion and implications: The UP5-levofloxacin conjugate demonstrated enhanced antimicrobial efficacy against resistant bacteria, with minimal hemolytic activity and favorable selectivity toward normal cells. This conjugate presents a promising approach to combating antimicrobial resistance, offering potential for improved therapeutic strategies.

Background and purpose: T-cell immunoglobulin and mucin-domain containing protein-3 (TIM-3)/galectin-9 (Gal-9)/ autocrine loop in myeloid leukemia stem cells provokes inflammation through the NF-κB signaling pathway, which is influential in the expression of inflammatory factors. Interleukin 1β (IL-1β) is a vital inflammatory cytokine that plays an important role in the proliferation and therapy resistance of acute myeloid leukemia (AML) cells. This study aimed to assess the effect of Gal-9 on IL-1β in the human leukemic U937 cell line.

Experimental approach: The U937 cells were cultured in different concentrations of Gal-9. Cell counting kit-8 was used to assess the effect of Gal-9 on human leukemic U937 cell proliferation. Also, its impact on the expression of TIM-3, Gal-9, IL-1β, IL-1βR, IL-1βRAP, and NLRP3 genes and IL-1β protein was studied by RT-PCR and ELISA, respectively. Moreover, the effect of Gal-9 on the NF-κΒ signaling pathway was evaluated by western blotting.

Findings/results: U937 cells were expanded in the presence of Gal-9 in a concentration-dependent manner. Following treatment of U937 cells with Gal-9, the gene expression of Gal-9, IL-1B, IL-1BR, and IL-1BRAP were significantly upregulated compared to the control group. The IL-1β concentration increased following Gal-9 treatment in a concentration-dependent manner, while following time its level significantly decreased. Furthermore, Gal-9 slightly increased NF-κΒ phosphorylation.

Conclusion and implications: Gal-9 increased IL-1β level as a critical inflammatory cytokine in the proliferation and resistance of AML cells to therapy. According to this finding, targeting and blocking the TIM-3/Gal-9 autocrine loop can suppress IL-Ιβ production and facilitate AML treatment.