Iranian Journal of Pharmaceutical Research最新文献

Background: Ellagitannins are well-recognized for their antioxidant, chemopreventive, anti-inflammatory, and neuroprotective efficacy. Due to their poor absorption and extensive catabolism, it is proposed that urolithins, as ellagic acid (EA) metabolites, are the real active molecules exerting these biological functions.

Objectives: This research evaluated the inhibitory effects of EA, urolithin A (Uro A), and urolithin B (Uro B) on the activity of recombinant human matrix metalloproteinase 9 (rhMMP-9). Dysregulation of MMP-9 activity is directly involved in various pathologies; therefore, inhibition of this enzyme has clinical importance.

Methods: The rhMMP-9 activity was measured by a standard protease assay with casein as the substrate in the presence and absence of natural compounds, and the corresponding kinetic parameters were calculated. Interaction affinity between the enzyme and each of the ellagitannins studied was determined by the surface plasmon resonance (SPR) method. Molecular docking was performed using the C-terminally truncated human pro-MMP-9 structure as the receptor protein (PDB ID 1L6J) to predict ligand-receptor interaction and visualize the in vitro results.

Results: The rhMMP-9 assay showed that EA, Uro A, and Uro B demonstrated inhibitory activity with IC₅₀ values of 17.14 µM, 33.29 µM, and 13.17 µM, respectively. Kinetic interaction parameters calculated using SPR analysis showed the lowest KD for Uro B (4.3 × 10^-5 M), compatible with its IC₅₀. KD values calculated were 11.3 × 10^-5 M for EA and 6.7 × 10^-5 M for Uro A. A mixed type of inhibition with a non-competitive-uncompetitive pattern for Uro A and Uro B and a competitive-non-competitive pattern for EA was revealed.

Conclusions: Our results showed the promising inhibitory potential of EA, Uro B, and Uro A to affect the catalytic activity of the MMP-9 enzyme and also confirmed the fibronectin domain as a potential site for drug design against MMP-9.

Background: Temporal lobe epilepsy (TLE) is a chronic neurological disorder characterized by hippocampal necrosis and apoptosis. Neuroinflammation plays a critical role in the pathophysiology of TLE, with toll-like receptor 4 (TLR4) serving as a key mediator. Activation of TLR4 leads to the release of pro-inflammatory cytokines, such as IL-6 and TNF-α, which contribute to neuronal injury and apoptosis. The TLR4 signaling pathway promotes neuroinflammation through nuclear factor kappa-B (NF-κB) activation, further exacerbating neuronal damage over time. Therefore, timely inhibition of TLR4 may help mitigate neuroinflammation and alleviate epilepsy symptoms.

Objectives: This study aimed to determine whether early inhibition of TLR4 can regulate seizures and apoptosis by targeting the NF-κB1 signaling pathway.

Methods: The TLR4 inhibitor C34 was administered intraventricularly to two experimental groups. The first group received the injection immediately after pilocarpine-induced seizures, while the second group was treated 24 hours post-pilocarpine injection. The expression levels of NF-κB1, TNF-α, and caspase-3 were analyzed using western blotting. Neuronal death in the hippocampus was assessed using hematoxylin and eosin (H&E) staining.

Results: The results demonstrated that early inhibition of TLR4 by C34, administered immediately after seizure induction, significantly reduced NF-κB1, TNF-α, and caspase-3 expression levels compared to the group that received C34, 24 hours later. Additionally, early treatment with C34 significantly prevented pilocarpine-induced neuronal death in the hippocampus compared to the late treatment group.

Conclusions: These findings highlight the importance of early intervention in reducing neuronal death and suppressing neuroinflammation in an epilepsy model. Inhibiting TLR4 immediately after seizure induction may serve as a potential therapeutic strategy to minimize inflammation-mediated neuronal damage in TLE. Further research is needed to explore the long-term effects of TLR4 inhibition in epilepsy treatment.

Background: Recent evidence has demonstrated the crucial role of macrophage polarization in promoting non-small cell lung cancer (NSCLC) progression within the tumor microenvironment.

Objectives: This study investigated the possible regulatory mechanism of macrophage polarization during NSCLC development.

Methods: The proportion of M1/M2 macrophages was examined by flow cytometry. The expression of macrophage markers and target molecules was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR), western blotting, and immunohistochemical staining. Non-small cell lung cancer cells were treated with conditioned medium (CM) from THP-1 macrophages. Cell counting kit-8 (CCK-8), scratch, and transwell assays were used to assess NSCLC cell growth and metastasis. Gene promoter activity was evaluated by dual-luciferase reporter assay. A xenograft model was adopted to determine NSCLC growth in vivo.

Results: Histone-lysine N-methyltransferase 2D (KMT2D) and integrin subunit alpha L (ITGAL) were lowly expressed in NSCLC tissues and cells. The KMT2D overexpression facilitated the polarization of macrophages from M2 to M1 type, which repressed the growth, migration, and invasion of NSCLC cells. Mechanistically, KMT2D promoted the transcription and expression of ITGAL. Inhibition of ITGAL abrogated KMT2D overexpression-mediated M1 macrophage polarization and its anti-cancer effects on NSCLC.

Conclusions: The KMT2D transcriptionally activated ITGAL to trigger M1 macrophage polarization, thereby delaying NSCLC progression. Our findings suggest KMT2D as a potential therapeutic target for NSCLC.

Background: The side effects and drug resistance associated with current antiepileptic drugs necessitate the design and synthesis of new candidate anticonvulsant agents.

Objectives: The present study aimed to design, synthesize, and screen a new series of gamma-aminobutyric acid (GABA) agonist derivatives for the treatment of seizures in an animal model.

Methods: The test chemical compounds were synthesized using known synthetic routes, and their structures were confirmed by various spectroscopic methods. Anticonvulsant activity was evaluated using the pentylenetetrazole (PTZ) animal model. Molecular docking studies were conducted to assess interactions with the GABA_A receptor.

Results: Some synthesized compounds significantly improved seizure symptoms and reduced mortality rates in the PTZ model. Derivative 3c demonstrated a correlation with the GABA_A receptor in the flumazenil test.

Conclusions: The synthesized molecules exhibited moderate to good activity compared to the control group. Derivative 3c notably increased seizure latency relative to the control. Flumazenil inhibitory effect tests indicated that 3c protects against PTZ-induced seizures via the synaptic GABA_A receptor.

Background: Aluminum (Al), lead (Pb), and mercury (Hg) are major environmental pollutants, and a large population may be simultaneously exposed to these metals. However, studies on the potential neurobehavioral effects of mixed exposure to Al, Pb, and Hg are lacking.

Objectives: This study aimed to evaluate neurobehavioral changes in mice following combined exposure to Al, Pb, and Hg and to investigate the effects of this exposure on dopaminergic neurotransmission within the striatum.

Methods: In this study, C57BL/6 mice (n = 10 per group) were assigned to control and metal-treated groups. Changes in motor coordination and locomotor activity that occurred when mice were simultaneously exposed to these metals via drinking water for 28 days were measured using the rotarod and open field tests. In addition, dopamine content and key factors involved in dopaminergic neurotransmission in the striatum were evaluated using real-time PCR and Western blot analysis.

Results: The mixed metal exposure decreased motor function and significantly reduced the content of dopamine in the striatum of the experimental mice (P < 0.001). Expression of tyrosine hydroxylase, vesicular monoamine transporter 2, and dopamine receptor D1, which are involved in dopaminergic neurotransmission in the striatum, was significantly decreased (P < 0.01), whereas expression of the dopamine transporter was significantly increased (P < 0.05). Dopamine receptor D2 expression was not significantly changed by the mixed metal exposure.

Conclusions: These results suggest that mixed exposure to Al, Pb, and Hg inhibits normal dopaminergic neurotransmission, resulting in neurobehavioral disorders.

Background: Quinazolinone derivatives have been documented to exhibit antidiabetic properties via the mechanism of dipeptidyl peptidase-4 (DPP-4) inhibition.

Objectives: To prepare and investigate the DPP-4 inhibitory activity in vitro and in silico of a series of novel 2-({2-[(dialkylamino)methyl]quinazolin-4-one-3-yl}methyl)benzonitrile derivatives.

Methods: The compounds were synthesized, and the chemical structures were confirmed through spectroscopic techniques. The in vitro DPP-4 inhibitory activity was assessed using an assay kit. Additionally, an in silico study was conducted using molecular docking methods to analyze the occurring binding interactions.

Results: The title compounds exhibited good inhibition against DPP-4 enzyme activity (IC₅₀: 1.4621 to 6.7805 µM). Among the compounds studied, the compound having morpholino-methyl substituted at C-2 (5d) exhibited the highest potency in DPP-4 inhibitory activity. Their activities were lower than sitagliptin as the reference standard with IC₅₀: 0.0236 µM and lead compound. In the in silico study, the compounds bound against the DPP-4 enzyme, with affinity values similar to those of sitagliptin. However, only compound 5f showed an interaction orientation and amino acid residues that were somewhat similar to those observed in the interaction between the DPP-4 enzyme and sitagliptin, as well as in the interaction between the DPP-4 enzyme and the lead compound.

Conclusions: A series of novel 2-({2-[(dialkylamino)methyl]quinazolin-4-one-3-yl}methyl)benzonitrile derivatives have been synthesized successfully. All the synthesized compounds had lower DPP-4 inhibitory activity than sitagliptin and the lead compound. The lower bioactivity was predicted due to the differences in the interaction between the synthesized and lead compounds against the DPP-4 enzyme.

Background: Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, remains a significant health concern due to its widespread prevalence and severe impact on immunocompromised individuals. Current treatments are limited, necessitating the exploration of new therapeutic agents.

Objectives: This study aimed to evaluate the efficacy and safety of 2-nitroimidazole as a potential treatment for toxoplasmosis in BALB/c mice, comparing its effects with the standard treatment, sulfadiazine.

Methods: In vitro assays were conducted to determine the half-maximal inhibitory concentration (IC50) of 2-nitroimidazole and sulfadiazine against T. gondii tachyzoites. The MTT assay was used to assess the cytotoxicity of 2-nitroimidazole on macrophages. In vivo experiments involved treating BALB/c mice infected with T. gondii with either 2-nitroimidazole or sulfadiazine, monitoring survival rates and therapeutic outcomes.

Results: In vitro results revealed IC50 values of 5.43 μM for 2-nitroimidazole and 2.99 μM for sulfadiazine, indicating potent anti-tachyzoite activity. The MTT assay showed that 2-nitroimidazole had low cytotoxicity, with significant cell viability even at higher concentrations. Based on the MTT assay findings, 40 μM of 2-nitroimidazole showed the highest level of toxicity towards macrophages. Furthermore, flow cytometry analysis revealed that this compound induced apoptosis in approximately 58.9% of tachyzoites. In vivo, all mice in the control group died by the eighth day. Treatment with sulfadiazine resulted in two mice surviving until the 14th day, while 2-nitroimidazole treatment saw one mouse surviving to the same day. These findings suggest that 2-nitroimidazole has comparable efficacy to sulfadiazine with potentially fewer side effects.

Conclusions: The study demonstrates that 2-nitroimidazole is a promising candidate for the treatment of toxoplasmosis, exhibiting strong anti-parasitic activity and low cytotoxicity. Further research is warranted to optimize dosing regimens and explore combination therapies to enhance its therapeutic potential.

Background: One of the most promising strategies to combat cancer is the use of immunotoxins.

Objectives: This study aimed to design two immunotoxins composed of antibody fragments against the EphA2 receptor, which is highly expressed in breast cancer.

Methods: EphA2-N-ricin and EphA2-C-ricin were designed by fusing scFv against the EphA2 receptor with the A chain of ricin in varying orders. mFold was used to analyze the mRNA stability of the constructs. The 2D and 3D protein structures of the constructs were predicted using prediction tools and verified by quality assessment tools. The physicochemical properties were calculated using ProtParam. Docking between the constructs and the EphA2 receptor was performed using HADDOCK software, and the 2D interaction plots of the complexes were generated using LigPlus. A 100 ns molecular dynamics (MD) simulation was conducted for docked complexes using Gromacs. Ultimately, the allergenicity and antigenicity of the constructs were determined.

Results: The designed immunotoxins had stable mRNAs, reliable 2D and 3D protein structures, and demonstrated high affinity and stable interactions with the receptor protein, as revealed by docking and MD analyses. Higher binding affinity and stability were observed for construct 2. Moreover, the designed immunotoxins lacked allergenicity and were identified as antigens.

Conclusions: Based on these observations, it is reasonable to conclude that both designed immunotoxins could serve as suitable immunotoxins; however, construct 2 exhibits more promising properties. Given these results, these immunotoxins could be used in empirical studies to treat breast cancer in vitro or in vivo.

Background: Bladder cancer (BC) is the most prevalent urogenital malignancy. Recently, the combination of natural compounds with chemotherapeutic agents has gained attention. Genistein, a natural flavonoid, exhibits anti-cancer properties and represents a promising candidate for treating various cancerous cells due to its cytotoxic potential and minimal adverse effects.

Objectives: This study aimed to evaluate the anti-cancer effects of genistein by regulating potential target genes and microRNAs involved in apoptosis and autophagy in EJ138 BC cells.

Methods: EJ138 BC cells were treated with different concentrations of genistein, and cell viability was assessed using the MTT assay. To determine the apoptotic rate of EJ138 BC cells following genistein treatment, flow cytometry with Annexin V/PI staining was performed. Additionally, real-time PCR was conducted to analyze the expression of miR-27a, miR-151, apoptotic genes (caspase-3, caspase-9), and autophagic genes (ATG12, Beclin1) after 48 hours of genistein treatment. Statistical analysis was carried out using SPSS V.22, with independent t-tests and one-way ANOVA. Results were considered statistically significant at P < 0.05.

Results: Our findings demonstrated that genistein inhibited the proliferation, growth, and viability of EJ138 BC cells and induced cell death. Real-time PCR results confirmed that genistein significantly upregulated miR-27a (P < 0.01), ATG12 (P < 0.01), Beclin1 (P < 0.05), caspase-3 (P < 0.001), and caspase-9 (P < 0.0001), while downregulating miR-151 expression (P < 0.05).

Conclusions: The results of this study suggest that genistein suppresses the proliferation and growth of human BC cells by modulating genes and microRNAs involved in apoptosis and autophagy. Therefore, genistein may serve as a novel therapeutic agent for BC treatment.

Background: Previous studies have demonstrated the efficacy of melatonin in alleviating symptoms of irritable bowel syndrome (IBS) and improving quality of life (QoL).

Objectives: Due to its superior bioavailability, this trial was designed to compare the effects of sublingual melatonin (SL melatonin) with a placebo in alleviating IBS symptoms, enhancing QoL, and addressing sleep disorders.

Methods: The IBS patients were randomly assigned to receive either 3 mg of SL melatonin or a matching placebo for eight weeks. Participants completed the IBS symptom severity score (IBS-SSS), IBS-quality of life 34 items (IBS-QoL 34), and Pittsburgh Sleep Quality Index (PSQI) questionnaires immediately before and after the study period.

Results: A total of 76 patients completed the trial over six months. The results indicated that the severity of IBS symptoms and QoL scores were significantly better in the SL melatonin group compared to the placebo group (P = 0.032 and P = 0.045, respectively). No participants withdrew from the trial due to serious side effects in either the SL melatonin or placebo groups.

Conclusions: Sublingual melatonin may be administered to IBS patients as a complementary treatment to alleviate symptoms and improve QoL.