Iranian Journal of Pharmaceutical Research最新文献

Background: The present study aimed to determine the pharmacokinetic parameters and bioequivalence of the test medicinal product, apixaban 5 mg tablet, and its reference product, Eliquis^®, in healthy male and female subjects under a fasted state.

Methods: Before in vivo evaluation, the quality control parameters of the products were evaluated and compared. This study was a single-dose, double-blind, 2-sequence, crossover, 2-period, randomized bioequivalence and pharmacokinetic study in 24 healthy individuals with a two-week washout period between doses. A series of blood samples were obtained over 48 hours after dose administration, and the samples were analyzed for their apixaban content using a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique. The pharmacokinetic parameters were computed using non-compartmental analysis.

Results: Both products passed the in vitro quality control criteria. Following administration of the apixaban tablet, the area under curve (AUC)_0-t, AUC_0-∞, and maximum plasma concentration (C_max) mean values for the test product were 1284.0 ng.h/mL, 1368.2 ng.h/mL, and 157.4 ng/mL, respectively, and for the reference product were 1310.6 ng.h/mL, 1406.5 ng.h/mL, and 157.6 ng/mL, respectively. The 90% confidence intervals (CI) of the geometric mean ratio for AUC_0-t (91.4 - 105.9), AUC_0-∞ (92.9 - 106.9), and C_max (87.1 - 101.9) fell within the predefined accepted range of 80% - 125%. No serious adverse events were observed.

Conclusions: The test product (apixaban 5 mg tablet) and reference product (Eliquis^®) achieved regulatory requirements for bioequivalence in healthy individuals under a fasted state.

Background: Wound healing and antibiotic resistance of pathogenic microbes have become global issues with serious consequences for the treatment of infectious diseases.

Objectives: The present study aimed to evaluate the antibacterial, anti-inflammatory, angiogenic, and wound healing properties of copper nanoparticles (CuNPs) synthesized using Lupinus arcticus extract.

Methods: The green synthesis was conducted using the precipitation method. The antibacterial activity of CuNPs against both methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MRSA) strains was evaluated. The effects of CuNPs on protein leakage, the expression levels of biofilm-related genes [e.g., intracellular adhesion A (icaA), intracellular adhesion D (icaD), and elastin-binding protein (EbpS) genes] in MRSA, as well as its impact on wound healing, angiogenesis, and anti-inflammatory effects, were assessed.

Results: The CuNPs exhibited a spherical shape with dimensions ranging from 10 to 85 nm. Both CuNPs alone and in combination with gentamicin (GNT) inhibited biofilm formation in MRSA, with minimum biofilm inhibitory concentration (MBIC₅₀) values of 6.6 µg/mL and 0.50 µg/mL for MRSA, respectively. The CuNPs significantly (P < 0.05) downregulated the expression levels of icaA, icaD, and EbpS in MRSA, particularly at half the minimum inhibitory concentration (1/2 MIC) and the minimum inhibitory concentration (MIC). Additionally, CuNPs markedly (P < 0.001) increased protein leakage in MRSA. The CuNPs demonstrated potent in vitro wound healing effects, promoting fibroblast cell proliferation and wound closure in a dose-dependent manner. Our results indicated a significant (P < 0.05) increase in the expression of HLA-G5 and VEGF-A genes in cells exposed to CuNPs. Furthermore, CuNPs reduced the expression levels of inflammatory genes in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells (P < 0.05).

Conclusions: The findings of this experimental test indicate that CuNPs, particularly in conjunction with GNT, exhibits promising antibacterial effects against MRSA without causing cytotoxicity to normal cells. This study also demonstrated that green-synthesized CuNPs possesses significant wound-healing properties through its antibacterial activity, inhibition of biofilm formation, induction of angiogenesis, and reduction of inflammation. However, further experiments are necessary to elucidate the precise mechanisms of action and potential toxicity of CuNPs.

Background: Breast cancer is among the most prevalent cancers in women and is the leading cause of mortality among women worldwide. Although a definitive cure for breast cancer remains elusive, essential fatty acids offer a promising therapeutic avenue.

Objectives: The present study aimed to synthesize 16 derivatives of docosahexaenoic acid (DHA) and linoleic acid (LA) and evaluate their anti-cancer properties in vitro.

Methods: Fourteen derivatives of LA and DHA were synthesized using a coupling method, while two ethylenediamine derivatives were synthesized via an ester intermediate. Molecular modeling was conducted using AutoDock Vina software. The cytotoxic effects of all compounds were assessed using the MTT assay on breast adenocarcinoma (MCF-7) cells. The mechanism of cell death induction by derivatives with the most favorable EC₅₀ values was determined through annexin V-FITC/PI flow cytometry analysis, focusing on early and late apoptosis.

Results: Docking results revealed that these compounds effectively interact with residues in the PTPB1 active site. All synthesized DHA and LA derivatives demonstrated cytotoxic effects on the MCF-7 cell line, with no significant cytotoxicity observed in normal human dermal fibroblasts (HDFs). Compounds D3 and L3, with EC₅₀ values of 15.96 ± 2.89 μM and 24.64 ± 1.81 μM, respectively, were identified as the most potent anti-cancer compounds among the derivatives.

Conclusions: The findings indicate that these functional fatty acid derivatives significantly reduce cancer cell viability. In addition to necrosis, compounds L3 and D3 induced apoptosis, with apoptosis rates of 20.5% and 47.1%, respectively.

Background: Technetium-^99m (^99mTc)-pyrophosphate (PYP) has been widely utilized in diagnosing bone disorders and certain cardiac conditions, such as amyloidosis, allowing for accurate imaging and detection of abnormalities within heart tissue. Rhenium, being in the same group as technetium in the periodic table, shares similar chemical properties. Rhenium-¹⁸⁸ (¹⁸⁸Re) possesses favorable nuclear properties for theranostic applications.

Objectives: This study focused on labeling sodium PYP with ¹⁸⁸Re and its biodistribution.

Methods: Different samples with varying amounts of PYP (5 - 22 mg), SnCl₂.2H₂O (0.2 - 6.0 mg), and ascorbic acid (0.3 - 7 mg) were prepared in vials. Initially, 0.08 mg of potassium perrhenate as a carrier in 1 mL saline was added to each vial. Subsequently, 370 - 3700 MBq of ¹⁸⁸ReO₄ ^- was added to the initial solution. The pH of the solutions was varied between 3 and 8. The compound was shaken vigorously for 30 seconds. Product incubation was performed in a secured container for 30 minutes at room temperature.

Results: Maximum labeling yield was achieved with 10 mg of PYP, 1 mg of SnCl₂.2H₂O, 0.3 mg of ascorbic acid, and 0.08 mg of potassium perrhenate as a carrier in 1 mL with 370 MBq of ¹⁸⁸ReO₄ ^- at pH 5. This compound showed good stability, and a radiochemical purity of 98.96% ± 0.1% was obtained. The biodistribution results of the radiolabeled ligand revealed that the maximum affinity for ¹⁸⁸Re-PYP was for bone after 4 hours, which was 2.24% ± 0.667% ID/g. The maximum uptake for the kidney, spleen, and liver was 1.53% ± 0.378%, 0.13% ± 0.086%, and 0.18% ± 0.12% ID/g, respectively.

Conclusions: The present study investigated the initial labeling efficiency of ¹⁸⁸Re-PYP along with its biodistribution and in vitro stability. The ¹⁸⁸Re-PYP conjugate, prepared under optimized conditions, demonstrated radiochemical purity and stability. The biodistribution of the compound in mice exhibited high affinity for bone, whereas the complex was eliminated through the kidneys.

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.