CYP3A4 Research Articles

The CYP4/20-HETE/GRP75 axis in the progression metabolic dysfunction-associated steatosis liver disease (MASLD) to chronic liver disease

IntroductionMetabolic-dysfunction-associated steatosis liver disease (MASLD) is a progressive liver disease from simple steatosis, steatohepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. Chronic liver diseases (CLDs) can lead to portal hypertension, which is a major cause of complications of cirrhosis. CLDs cause structural alterations across the liver through increased contents of extracellular matrix (ECM), driving dysfunction of liver sinusoidal endothelial cells (LSECs) alongside hepatic stellate cells (HSCs) and activated resident or infiltrating immune cells. Bioactive arachidonic metabolites have diverse roles in the progression of MASLD. Both secreted levels of 20-hydroxyeicosatetraenoic acid (20-HETE) and epoxyeicosatrienoic acid (EET) are elevated in patients with liver cirrhosis.MethodsCLD samples were evaluated for changes in free fatty acids (FFA), cholesterol, bilirubin, bile acid, reactive oxygen species (ROD), lipid peroxidation, myeloperoxidase activity and hydroxyproline levels to evaluate the degrees of liver damage and fibrosis. To address the role of the CYP4/20-HETE/GPR75 axis, we measured the amount and the synthesis of 20-HETE in patients with CLD, specifically during the progression of MASLD. Additionally, we evaluated gene expression and protein levels of GPR75, a high-affinity receptor for 20-HETE across CLD patient samples.ResultsWe observed an increase in 20-HETE levels and synthesis during the progression of MASLD. Increased synthesis of 20-HETE correlated with the expression of CYP4A11 genes but not CYP4F2. These results were confirmed by increased P4504A11 protein levels and decreased P4504F2 protein levels during the development and progression of MASLD. The gene expression and protein levels of GPR75, the major receptor for 20-HETE, increased in the progression of MASLD. Interestingly, the CYP4A11 and GPR75 mRNA levels increased in steatohepatitis but dramatically dropped in cirrhosis and then increased in patients with HCC. Also, protein levels of P4504A11 and GPR75 mirrored their mRNA levels.DiscussionThese results indicate that the CYP4A11 and subsequent GPR75 genes are coordinately regulated in the progression of MASLD and may have multiple roles, including 20-HETE activation of peroxisome proliferator-activated receptor α (PPARα) in steatosis and GPR75 in CLD through either increased cell proliferation or vasoconstriction in portal hypertension during cirrhosis. The abrupt reduction in CYP4A11 and GPR75 in patients with cirrhosis may also be due to increased 20-HETE, serving as a feedback mechanism via GPR75, leading to reduced CYP4A11 and GPR75 gene expression. This work illustrates key correlations associated with the CYP4/20-HETE/GPR75 axis and the progression of liver disease in humans.

P-11 A CASE OF TUBERCULOSIS ADRENAL INSUFFICIENCY

Abstract Introduction Adrenal tuberculosis is rare form of extrapulmonary tuberculosis, and may cause to primary adrenal insufficiency. This report presents a case of middle age male diagnosed with adrenal tuberculosis. Tuberculosis is a major health concern worldwide. The extrapulmonary manifestations account for 5% to 15% of all cases, although adrenal tuberculosis is uncommon, it can lead to primary adrenal insufficiency and has diagnostic challenges due to its nonspecific presentation. Pulmonary Tuberculosis is still common in Iraq, although no case reports exit up to our knowledge of adrenal Tuberculosis. Clinical Case 40-year-old man, sanitation worker, presents with 3 months history of significant weight loss and fever with night sweating. He complains of nausea, occasional vomiting, fatigue, unintentional weight loss of approximately 12 kg, decreased appetite, abdominal pain and joint pain. He also reports skin hyperpigmentation particularly in the hand creases, mouth and mucous membranes of the lips. There are no respiratory symptoms such as shortness of breath or cough. The past medical history is unremarkable. There is positive family history of pulmonary tuberculosis (grandmother). He does not smoke or drink alcohol. Physical examination reveals ill-looking malnourished patient, with BP of 100/60 with postural hypotension, PR of 80 bpm and RR of 24/min. There is diffuse hyperpigmentation particularly noted in palmer creases and oral cavity. The other system examinations are normal. Laboratory investigations revealed mild normochromic normocytic anemia (Hb 11.4), and mild lymphocytosis, Na 126 mEq/l, k 4.1 mEq/l, FBS 87 mg/dl, high ESR, mild elevation of TSB 1.8 mg/dl normal renal function, morning cortisol of 0.75 Mcg/dl and morning ACTH of 1808 pg/ml. Imaging studies show a normal chest X-ray. Abdominal CT reveals bilateral adrenal enlargement in size with lobulated irregular contours and heterogeneous density exhibiting foci of calcification and central cystic/necrotic non-enhancing centers, with predominantly peripheral enhancement seen on the contrast scan. The mean density of the solid parts of the glands is around 30-40 HU on the native scan, the right adrenal measures 2.7x1.7x2.7 cm, the left adrenal measures 5.6x3.2x2.5 cm. IGRA test is positive. Adrenal biopsy was not performed because the presentation was highly suggestive of adrenal tuberculosis.The diagnosis was primary adrenal insufficiency due to adrenal tuberculosis Anti TB drugs were initiated on standard regimen of INH, Rifampicin, Pyrazinamide, and Ethambutol. Hydrocortisone replacement was initiated. The patient required higher dose of hydrocortisone (20 mg in the morning, 10 mg at 6 pm) which could be due to CYP 3A4 induction by rifampicin Fludrocortisone 0.1mg daily was also started. The patient’s symptoms improve dramatically after initiation of treatments, with gradual increase in body weight.Figure 1:Hyperpigmentation of the patient hands

Origanum vulgare and hemorrhagic risk, about a case

The use of medicinal plants in Algeria is an ancestral practice that remains relevant today. The population relies on plants to treat various diseases and everyday ailments, which can be dangerous, especially when taking medication [1,2]. The interaction between plants and medication can lead to a modification of the plasma concentrations of the latter, which can impact its therapeutic effectiveness and be responsible for toxicity or therapeutic failure [1,2]. The interaction can also be pharmacodynamic, resulting in the potentiation of the effect or its antagonism. The risk is even higher when the medication has a narrow therapeutic range or presents high inter- or intra-individual variability, such as with vitamin K antagonist anticoagulants. We report the case of patient B.Z., aged 77, who is suffering from complete atrial fibrillation (ACFA). The patient has been under antivitamin K (AVK) treatment with acenocoumarol for 8 years and has been followed at the central laboratory of Didouche Mourad Hospital since August 2019. The patient has been stable with an INR within the therapeutic target range of 2-3 for 4 years. She is compliant and adheres to the dietary regimen. The dosage of acenocoumarol is alternated between 1/2 tablet and 1/4 tablet. On September 6, 2023, during a routine check-up, the patient's INR was 6.42, and the PT was 17.9%. The recommendation was to stop AVK and perform a control the following day. The interview revealed that the patient had started taking an infusion of oregano (Origanum vulgare), and occasionally verbena, at a rate of one 200 ml cup per day for a week. On September 7, 2023, the INR decreased to 3.80 and the PT to 26.9%, so the recommendation was to resume AVK at a dosage of 1/4 tablet to 1/2 tablet, with discontinuation of the herbal tea intake and an INR check after 72 hours. On the fourth day after resuming the anticoagulant, the INR was 2.5. Despite being informed of the risk of interaction, the patient resumed taking the freshly picked oregano herbal tea from her garden. Given the patient's determination to continue her infusion, the recommendation was made to limit her intake, to prepare much more diluted infusions, and to reduce the dose of acenocoumarol to 1/4 tablet; this allowed the stabilization of the patient's INR. The last check-up was performed on April 7, 2024, with an INR value of 2.3. Oregano, a plant from the Lamiaceae family, is used in herbal tea to treat a wide range of diseases, including joint pain, respiratory conditions, and digestive disorders. Similarly, verbena is also used to combat sleep disorders [2-4]. Origanum and verbena are both plants rich in polyphenols, as well as tannins and flavonoids. These substances are inhibitors of CYP 450, particularly CYP 2C9 and CYP 3A4, which are responsible for the metabolism of many drugs, including painkillers, antihypertensives, central nervous system drugs, and anticoagulants such as acenocoumarol. Origanum also exhibits anticoagulant activity due to its high content of carvacrol and thymol [4-6]. These two modes of interaction lead to an increase in the pharmacodynamic effect of acenocoumarol and, therefore, an increased risk of bleeding. This was the case for our patient, who saw her INR increase to more than twice the normal level, coinciding with the intake of oregano and verbena infusions, which normalized after their discontinuation. Oregano is an endemic plant widely used in Algeria, whose anti-inflammatory properties are well established. However, its side effects, toxicity, and risk of interaction when combined with medications are less known. The interaction between oregano and AVK (acenocoumarol) exists and deserves to be recognized and monitored, especially in a population with a traditional culture where the use of plants is common. This practice should be accompanied by national phytovigilance.

CYP3A4-Mediated Bioactivation of the β-Amyloid Precursor Protein-Cleaving Enzyme 1 Inhibitor JNJ-54861911 Results in Redox-Neutral Addition of Glutathione via Catalysis by Glutathione S-Transferase α1, Identified as the Major Target Protein in Human Hepatocytes.

The β-amyloid precursor protein-cleaving enzyme 1 (BACE1) inhibitor JNJ-54861911, a candidate for the treatment of Alzheimer's disease, was withdrawn from clinical trials due to drug-induced liver injury (DILI). This paper describes our investigation of the metabolism of JNJ-54861911 to understand the potential contribution to the observed DILI. In human hepatocytes, JNJ-54861911 is metabolized by CYP450 3A4 to a reactive intermediate (RI), which undergoes glutathione (GSH) addition at C6 of the 2-amino-4-methyl-1,3-thiazin-4-yl moiety via glutathione S-transferase α1 (GSTA1) catalysis. Despite the preponderant role of CYP3A4 as an enabler, the adduct has the same level of oxidation as that of JNJ-54861911. The exact mechanism of RI formation might involve a sulfoxide (with further reduction) or tautomeric forms of JNJ-54861911 bearing a reactive thiazinium cation activating both the C2 and C6 positions. The cell pellet from the human hepatocyte incubated with 14C-JNJ-54861911 was analyzed via gel electrophoresis, resulting in the identification of a major protein adduct on GSTA1, a cross-link resulting from the addition of GSH and lysine 120 to JNJ-54861911, most likely on position C6 and on the nitrile, respectively. Ultimately, this major adduct might only account for 15-25% of the total covalent binding (CVB). Other important contributors to CVB might exist, like the bioactivation of the major diaminothiazine metabolite (DIAT). The level of covalent binding (CVB) burden (fraction of the dose resulting in CVB) is clearly below 1 mg/day, with a low daily dose of 25 mg. Despite this limited magnitude of CVB, it could still contribute to the liver enzyme elevations observed in approximately 20% of the patients and to the few cases of severe immune-mediated DILI. The latter could occur through a haptenization phenomenon and/or by inducing stress in hepatocytes. Such stress may activate an innate immune response, which, in turn, promotes the adaptive immune response.

Insights into CYP 1A2 dynamics in acute lymphocytic leukemia: Gene regulation, promoter methylation, and pesticide exposure in the Kashmiri population

Insights into CYP 1A2 dynamics in acute lymphocytic leukemia: Gene regulation, promoter methylation, and pesticide exposure in the Kashmiri population

Structure identification of myricetin-phenylacetaldehyde adducts and their potential biological activities

Our previous research discovered that myricetin could effectively inhibit the formation of heterocyclic aromatic amines (HAAs) in cantonese baked foods by trapping phenylacetaldehyde to form adducts. However, the structure and biological activity of these adducts were still unknown. In this study, we identified two myricetin-phenylacetaldehyde adducts from cantonese mooncakes, BYQ-2 and BYQ-3, using pre-HPLC. These adducts were found to be the products of phenylacetaldehyde addition at the C-8 and C-6 positions of myricetin, followed by cyclization with hydroxyl groups at the C-7 and C-5 positions. Antioxidant assays revealed that BYQ-2 and BYQ-3 have stronger radical scavenging abilities than myricetin in the concentration range of 12.5 ∼ 800 μg/mL in the three samples. Additionally, both adducts showed potential health benefits by inhibiting α-glucosidase and CYP450 1A2, enzymes involved in blood sugar regulation and HAAs metabolism. BYQ-2 exhibited the highest inhibitory potency against α-glucosidase and CYP450 1A2 with IC50 values of 10.32 μg/mL and 5.44 μg/mL, respectively. Theoretical calculations suggested that hydrogen bonding and hydrophobic interactions are the primary forces driving enzyme binding, with BYQ-2 showing the highest binding energies (−7.75 kcal/mol for α-glucosidase and −9.31 kcal/mol for CYP450 1A2). Our findings suggested that myricetin inhibited HAAs in baked foods while also enhancing food safety and providing health benefits through its adduction with small molecule aldehydes. In future research, it is necessary to further evaluate the absorption and metabolic behavior as well as safety of myricetin-HAAs active intermediate adducts at the cellular and animal experimental level.

Triterpenoids from Ganoderma lucidum inhibit cytochrome P450 enzymes interfering with the metabolic process of specific clinical drugs.

Ganoderma lucidum (G. lucidum), which possesses various biological effects, has been widely used as traditional medicine and functional food in Asian countries, especially China. In consideration of its various biological effects on human healthcare, G. lucidum was usually used in combination with other drugs. However, the potential drug-drug interaction induced by G. lucidum through cytochrome P450 enzymes (CYPs) remain unknown. Using the in vivo activity assay of CYPs, the inhibitory effects of G. lucidum and its constituents could be evaluated. The interference of G. lucidum on the metabolic processes of clinical drugs could be investigated. The chemical constituents of G. lucidum could be identified using LC-MS. The interaction between bioactive compounds and CYPs could be proposed through in silico docking analysis and molecular dynamics. The dichloromethane extract of G. lucidum could inhibit various CYP450 subtypes in vitro and interfere with the pharmacokinetics of four drugs in rats. Triterpenoids were identified as the main constituents of the dichloromethane extract by Q-TOF-MSn in preliminary analyses. Then, a triterpenoid library containing 66 compounds was established to evaluate their inhibitory effects against CYP 1A2, 2D6, 3A4, 2A6, 2B6, 2C9, and 2C19. CYP 1A2 was inhibited by most lanostane triterpenoids, indicating a strong affinity for these compounds. Among triterpenoids, compound 25 displayed a broad inhibitory effect against CYPs, except for CYP 3A4, 2D6, 2C9, and 2C19. Finally, compounds 6 and 25 exhibited interference with the metabolism of 16 drugs through the inhibition of CYPs in vitro. In silico molecular docking analyses for assaying the interaction between compound 25 and CYPs indicated that the hydrogen bonds formed between the hydroxyl groups and amino acid residues. G. lucidum displayed broad inhibitory effects on CYPs, with triterpenoids as the main bioactive constituents, which may induce potential drug-drug interaction. This information should be helpful for the rational use of G. lucidum in promoting human health.

Clozapine Toxicity Predictor: Deep neural network model predicting clozapine toxicity and its therapeutic dose range

Clozapine is the gold standard for treatment-resistant schizophrenia; however, its superior efficacy is accompanied by potentially serious adverse events (neutropenia, seizures, constipations, pneumonia), many of which are also concentration-dependent. As such, clozapine dose titration should be guided by therapeutic drug monitoring (TDM). However, access to TDM is often limited. The present study describes a new deep neural network that can predict the concentrations, toxicity and therapeutic dose range for clozapine and norclozapine. The model was trained on basic clinical data (biological sex, age, clozapine daily dose, BMI, CRP and number of CYP 1A2 and 3A4 substrates, inhibitors and inducers) from 69 patients treated with different clozapine doses. Our findings provide the training efficacy data for the model, as well as an analysis of clozapine and norclozapine blood concentrations in a test group of three additional patients, to demonstrate its practical capabilities. The model is licensed on a free and permissive 2-Clause BSD license and is available to all clinicians; it can be accessed as a web application, available at https://csk.umed.pl/clotop.

A Computational Pipeline Observes the Flexibility and Dynamics of Plant Cytochrome P450 Binding Sites.

Binding site flexibility and dynamics strongly affect the ability of proteins to accommodate substrates and inhibitors. The significance of these properties is particularly pronounced for proteins that are inherently flexible, such as cytochrome P450 enzymes (CYPs). While the research on human CYPs provides detailed knowledge on both structural and functional level, such analyses are still lacking for their plant counterparts. This study aims to bridge this gap. We developed a novel computational pipeline consisting of two steps. Firstly, we use molecular dynamics (MD) simulations to capture the full conformational ensemble for a certain plant CYP. Subsequently, we developed and applied a comprehensive methodology to analyze a number of binding site properties-size, flexibility, shape, hydrophobicity, and accessibility-using the fpocket and mdpocket packages on MD-generated trajectories. The workflow was validated on human CYPs 1A2, 2A6, and 3A4, as their binding site characteristics are well known. Not only could we confirm known binding site properties, but we also identified and named previously unseen binding site channels for CYPs 1A2 and 2A6. The pipeline was then applied to plant CYPs, leading to the first categorization of 15 chosen plant CYPs based on their binding site's (dis)similarities. This study provides a foundation for the largely uncharted fields of plant CYP substrate specificity and facilitates a more precise understanding of their largely unknown specific biological functions. It offers new insights into the structural and functional dynamics of plant CYPs, which may facilitate a more accurate understanding of the fate of agrochemicals or the biotechnological design and exploitation of enzymes with specific functions. Additionally, it serves as a reference for future structural-functional analyses of CYP enzymes across various biological kingdoms.

Liver CYP4A autophagic-lysosomal degradation (ALD): A major role for the autophagic receptor SQSTM1/p62 through an uncommon target interaction site.

The hepatic P450 hemoproteins CYPs 4A are typical N-terminally anchored Type I endoplasmic reticulum (ER)-proteins, that are inducible by hypolipidemic drugs and other "peroxisome proliferators". They are engaged in the ω-/ω-1-oxidation of various fatty acids including arachidonic acid, prostaglandins and leukotrienes and in the biotransformation of some therapeutic drugs. Herein we report that of the mammalian liver CYPs 4A, human CYP4A11 and mouse Cyp4a12a are preferential targets of the ER-lysosome-associated degradation (ERLAD). Consequently, these proteins are stabilized both as 1%Triton X100-soluble and -insoluble species in mouse hepatocytes and HepG2-cells deficient in the autophagic initiation ATG5-gene. Although these proteins exhibit surface LC3-interacting regions (LIRs) that would target them directly to the autophagosome, they nevertheless interact intimately with the autophagic receptor SQSTM1/p62. Through structural deletion analyses and site-directed mutagenesis, we have identified the Cyp4A-interacting p62 subdomain to lie between residues 170 and 233, which include its Traf6-binding and LIM-binding subdomains. Mice carrying a liver-specific genetic deletion of p62 residues 69-251 (p62Mut) that includes the CYP4A-interacting subdomain also exhibit Cyp4a-protein stabilization both as Triton X100-soluble and -insoluble species. Consistently, p62Mut mouse liver microsomes exhibit enhanced ω- and ω-1-hydroxylation of arachidonic acid to its physiologically active metabolites 19- and 20-HETEs relative to the corresponding wild-type mouse liver microsomes. Collectively, our findings suggest that any disruption of CYP4A ERLAD results in functionally active P450 protein and consequent production of proinflammatory metabolites on one hand, and insoluble aggregates on the other, which may contribute to pathological aggregates i.e. Mallory-Denk bodies/inclusions, hallmarks of many liver diseases.

In Vivo Chemosuppressive Effects of Kolaviron on 7,12-Dimethylbenzanthracene-Induced Mammary Lesions are Associated with Changes in Levels of Estrogen Receptor-α, CYP 1A1, Proinflammatory Cytokines, and Alterations to Metabolic Pathways Implicated in Mammary Carcinogenesis.

Garcinia kola is a medicinal food commonly consumed in Sub-Sahara Africa, for which Kolaviron (KV) is the active portion. As a follow-up to our earlier chemopreventive studies, we investigated the chemotherapeutic effects of KV on experimentally induced mammary carcinogenesis in female Wistar rats. Mammary carcinogenesis was induced using 80 mg/kg of 7,12-dimethylbenzanthracene (DMBA) administered by oral gavage. One hundred-fifty days post-DMBA induction, estrogen receptor-α (ER-α) levels were determined in the experimental rats before treatment with KV commenced. Treatment was done using 50, 100, and 200 mg/kg KV thrice a week for 4 weeks, after which the experiment was terminated. Significantly higher levels of estrogen receptor-α, CYP 1A1, malondialdehyde, formation of lobular neoplastic cells, epithelial hyperplasia, lymphocyte infiltration, and increased cytokine (interleukin-6 and tumor necrosis factor-α) activity were observed in DMBA-induced rats, which were attenuated in KV-treated rats. Tyrosine metabolism was exclusively enriched in DMBA-induced rats in contrast to KV-treated rats. Collectively, the results point to the chemotherapeutic potential of KV.

In vitro and in vivo preclinical pharmacokinetic characterization of aficamten, a small molecule cardiac myosin inhibitor

Aficamten, a small molecule selective inhibitor of cardiac myosin, was characterised in preclinical in vitro and in vivo studies. Protein binding in human plasma was 10.4% unbound and ranged from 1.6% to 24.9% unbound across species. Blood-to-plasma ratios ranged from 0.69 to 1.14 across species. Aficamten hepatic clearance in human was predicted to be low from observed high metabolic stability in vitro in human liver microsomes. Aficamten demonstrated high permeability in Caco-2 cell monolayers. Aficamten in vivo clearance was low across species at 8.8, 2.1, 3.3, and 11 mL/min/kg in mouse, rat, dog, and monkey, respectively. The volume of distribution was low-to-high ranging from 0.53 in rat to 11 L/kg in dog. Oral bioavailability ranged from 41% in monkey to 98% in mouse. Aficamten was metabolised in vitro to eight metabolites with hydroxylated metabolites M1a and M1b predominating. CYP phenotyping indicated multiple CYPs (2C8, 2C9, 2D6, and 3A4) contributing to the metabolism of aficamten. Human clearance (1.1 mL/min/kg) and volume of distribution (6.5 L/kg) were predicted using 4-species allometry employing ‘rule-of-exponents’. A predicted 69 hour half-life is consistent with observed half-life in human Phase-1. No CYP-based drug-drug interaction liability as a precipitant was predicted for aficamten.

Assessing the impact of aprepitant on response to dose-intensive cyclophosphamide, etoposide, and cisplatin (DICEP).

Despite evidence demonstrating the effectiveness of aprepitant for chemotherapy-induced nausea and vomiting (CINV), its use in stem cell transplant settings across Canada is not standard. While pharmacokinetic data exists, the clinical significance of cytochrome P450 3A4 (CYP 3A4) inhibition of cyclophosphamide by aprepitant is unclear. Reduced activation of cyclophosphamide may reduce the effectiveness of dose-intensive cyclophosphamide, etoposide, and cisplatin (DICEP). To compare response rates to DICEP in patients with Hodgkin lymphoma (HL) and diffuse large B-cell lymphoma (DLBCL) in the presence and absence of aprepitant. A retrospective review of patients who received full-dose DICEP for relapsed/refractory HL or DLBCL between June 1995 and September 2018 at the Foothills Medical Centre (FMC) in Calgary, Alberta, Canada was conducted. Descriptive statistics were used to assess response rate, as defined by the 2007 International Working Group response criteria. Of the 218 patients included in this study, 87.6% of patients in the control group and 88.5% of patients in the aprepitant group responded to DICEP (difference 0.025 [95% CI, -0.066 to 0.114], p = 0.827). Univariate analyses for age, sex, type of cancer, stage of cancer, number of prior relapses, and relapse status were not significant. No significant differences were observed for secondary outcomes. Response rates to DICEP in relapsed/refractory HL and DLBCL patients were similar regardless of aprepitant use. Considering these results and the effectiveness of aprepitant in CINV, its addition to standard antiemetic therapy in patients receiving DICEP should be given strong consideration in the transplant setting.

Assessment of Nutritional Status of Children aged 1-5 years attending the Outdoor Patients Department of Sheikh Zayed Hospital, Rahim Yar Khan, Pakistan

Background: Cytochrome P450 enzymes play vital roles in metabolizing drugs, endogenous compounds, and environmental pollutants. Among them, Cytochrome P450 1A2 (CYP1A2), CYP1A1 and CYP1B are particularly important for activating carcinogens. Computational modeling of CYP1A2 is essential for understanding its interactions with various molecules, substrates, and inhibitors. Objective: To characterize the structure of CYP1A2 and explore the binding of alpha-naphthoflavone to its active site. Study Design: In Silico study (Computational modeling). Place of Study: International Center of Medical Sciences Research (ICMSR), Islamabad, Pakistan. Material and Methods: Using the Swiss PDB Viewer, the structural features of CYP1A2 were assessed, focusing on key residues, motifs, helices, and conserved regions. Results: Our findings identified specific binding sites for Alpha-Naphthoflavone (ANF), highlighting its potential as a potent inhibitor of CYP1A2. This research contributes to our knowledge of the clinical and toxicological implications associated with CYP1A2. Conclusion: Structural differences were found between CYP1A2 and related enzymes, with less than 40% sequence identity compared to several other P450s. The study predominantly compares CYP1A2 with CYP 2A6 and CYP3A4 due to these differences. T he developed structural models offer a fast and precise method for studying CYP1A2, aiding in understanding its role in drug metabolism and toxicology.

Metabolic profiling of a new synthetic cannabinoid receptor agonist, ADMB-FUBIATA, with human liver microsomes, human primary hepatocytes and human recombinant CYP450 enzymes using LC-quadrupole-orbitrap MS

A novel synthetic cannabinoid receptor agonist (SCRA), ADMB-FUBIATA, featuring an acetamide-linked structure, has emerged on the illicit drug market. To provide dependable verification of its consumption and identify reliable biomarkers, we investigated an in vitro metabolism study of ADMB-FUBIATA incubated with human primary hepatocytes (HPHs) for the first time and correlated our findings with those from human liver microsomes (HLMs). In this work, ADMB-FUBIATA (10 μM) was incubated with HLM and HPH for 1 and 5 h, respectively, and then subjected to LC-quadrupole-orbitrap MS. A total of 25 metabolites across 8 metabolic pathways were identified after incubation with HLM and HPH, respectively. Monohydroxylation and N-dealkylation were the major metabolic pathways, and formation to ketone was first identified. In addition, the metabolism of ADMB-FUBIATA were found to be mediated by multiple CYP450 enzymes, predominantly CYP2C19, 2D6, and 3A4. This research also initially characterized the fragmentation patterns of the metabolites of ADMB-FUBIATA, elaborating on their structural relationship with ADMB-FUBIATA analogs. To effectively monitor ADMB-FUBIATA abuse, metabolites M4 and M1 were proposed as reliable biomarkers by cross-validating the HLM and HPH incubation results.

An Unusual Case Report of Iatrogenic Metrorrhagia and Hematochezia Following Itraconazole

Abstract Itraconazole is an effective triazole antimycotic drug that has a desirable pharmacokinetic profile to combat various fungal pathogens. It acts by inhibiting ergosterol biosynthesis in fungal cell membranes. Itraconazole is not totally selective toward Cytochrome P450 family 3 subfamily A member 4 (CYP450 3A4), but exhibits the most potent inhibition of CYP450 3A4. Thus, these drugs interfere with CYP450 3A4 proteins expressed in the liver, intestine, kidney, adrenals, ovary, and testis. We present a case report of metrorrhagia and hematochezia following intake of itraconazole, a commonly used azole drug against dermatophytic infection. This article strongly highlights eliciting menstrual history and gynecological- and surgical-related problems before prescribing itraconazole. Also, to be aware of the vaginal bleeding being the unusual side effects of itraconazole.

Pattern of CYP3A5 and MDR-1 single nucleotide polymorphism and its impact on Tacrolimus levels and clinical outcomes in living renal allograft recipient

Abstract Introduction Renal transplant is best form of renal replacement therapy. Most favored immunosuppression includes Tacrolimus, mycophenolate mofetil and steroids. Tacrolimus has narrow therapeutic index and requires therapeutic drug monitoring (TDM). However, there is wide variation in tacrolimus level with weight based fixed dosage regimens. This variability is due to polymorphism of major pathways of metabolism ie CYP3A5 and MDR1 genes. Fast metabolizer require higher dosage and slow metabolizer require lower dosage. Genotype based dosing strategy may be useful to achieve early therapeutic level and reduce infections and rejections. Methodology 160 transplant patients at tertiary care hospital in India were included in this study from 2016 to 2018. Genetic polymorphism analysis in CYP3A5 and MDR1 gene was carried out at time of transplant. All patients were given a fixed weight-based dosage of Tacrolimus. Data was analyzed in relation to genotype polymorphism. Results and discussion 69.2% of Wild variants of CYP 3A5 (Fast metabolizers) have low initial tacrolimus levels. 51.5% of Homo variants (Slow metabolizers) have high initial tac levels. However, all variants achieve optimum tacrolimus levels at same time (mean 12.4 days). There were higher number of infections among slow metabolizers. Conclusion A fixed dosing regimen with TDM result in high and low initial tacrolimus levels in slow and fast metabolizers respectively and more infections in slow metabolizers. However, graft rejections being fewer in number, were not different. A larger sample with genotype based dosing is required to test such a strategy.

A Scoping Review on Recent Progress on Purpurin and its Derivatives

Background: Purpurin is being used as a red dye for many decades. But recently, due to its pharmacological properties, purpurin and its derivatives have attracted a lot of researchers for the treatment of various ailments, such as cancer, Alzheimer’s disease, depression, etc. Objectives: The objective of this study is to provide an overview of its pharmacological properties, pharmacokinetic studies, synthesis, isolation, quality assurance, and patent studies. Method: A systemic scoping review was undertaken. Three databases (Pubmed, Scopus, and Google Scholar) and patent websites were searched using relevant words (e.g., purpurin, purpurin derivatives, anticancer, toxicity, etc.). All outcomes for studies that met the inclusion criteria were included in the review. Extracted data were accumulated using tables, figures, and accompanying narrative descriptive summaries. The review was reported using the preferred reporting items for scoping review (PRISMAScR) guidelines. Sixty-eight studies and eighty-six patents met the inclusion criteria, mostly preclinical (in vitro, in vivo, and in silico) studies performed in rats, mice, dogs, and zebrafish, followed by one clinical trial study. Results: The potent antioxidant nature of purpurin is the main reason behind its vast pharmacological properties. It acts by decreasing mitochondrial stress and by acting on the endoplasmic reticulum. It also crosses the BBB barrier, has high GI absorption, and follows the Lipinski rule, which makes it a good drug for various neurodegenerative disorders. It inhibits various CYP-450, CYP 1A2, and CYP 3A4 enzymes, which are responsible for causing mutations. It gets photosensitized by UV light and causes ROSdependent apoptosis in cancer cells. Conclusion: This scoping review highlights purpurin and its derivatives as highly prized moieties in the treatment of various neurological conditions and cancer. The unique nature of purpurin is responsible for its pharmacological properties, which are due to the presence of hydroxyl and keto groups at specific positions. It gets photosensitized by UV and laser light and acts as an anticancer drug. But the lack of robust evaluation in clinical studies is another major concern. Purpurin can be seen in the prescription in the future, although a lot of work still needs to be done.

Identification of New Substrates and Inhibitors of Human CYP2A7.

CYP2A7 is one of the most understudied human cytochrome P450 enzymes and its contributions to either drug metabolism or endogenous biosynthesis pathways are not understood, as its only known enzymatic activities are the conversions of two proluciferin probe substrates. In addition, the CYP2A7 gene contains four single-nucleotide polymorphisms (SNPs) that cause missense mutations and have minor allele frequencies (MAFs) above 0.5. This means that the resulting amino acid changes occur in the majority of humans. In a previous study, we employed the reference standard sequence (called CYP2A7*1 in P450 nomenclature). For the present study, we created another CYP2A7 sequence that contains all four amino acid changes (Cys311, Glu169, Gly479, and Arg274) and labeled it CYP2A7-WT. Thus, it was the aim of this study to identify new substrates and inhibitors of CYP2A7 and to compare the properties of CYP2A7-WT with CYP2A7*1. We found several new proluciferin probe substrates for both enzyme variants (we also performed in silico studies to understand the activity difference between CYP2A7-WT and CYP2A7*1 on specific substrates), and we show that while they do not act on the standard CYP2A6 substrates nicotine, coumarin, or 7-ethoxycoumarin, both can hydroxylate diclofenac (as can CYP2A6). Moreover, we found ketoconazole, 1-benzylimidazole, and letrozole to be CYP2A7 inhibitors.

Assessment of microplastic-contaminated liver through gene expression profiling of four commercial fish species in the Lagos Lagoon, Nigeria

Microplastic (MP) occurrence in the Lagos Lagoon has been on a progressive increase in recent years. The potential impacts and hazards of MPs on commercially available fish species from the Lagoon have been understudied. This study aims to assess molecular damage due to microplastic accumulation in the liver of four commercial fish species (Oreochromis niloticus, Chrysichthys nigrodigitatus, Clarias gariepinus, and Gymnarchus niloticus) sourced directly from the Lagoon. The results revealed that MP load in the liver of the fish species ranged between 7.2 ± 1.9 and 9.5 ± 4.4 particles/individual, with G. niloticus recording the maximum concentration. The observed extracted MPs are predominantly black and blue fibres while the Fourier Transform Infrared spectroscopy revealed polyethylene dominance in the plastic polymers. Relative gene expression results revealed statistically significant downregulation of the CYP 1a gene and upregulation of the HSP 70 gene in all the microplastic-laden liver samples except in G. niloticus. The findings from this study suggest that microplastic contamination in fish tissues may exacerbate cellular toxicity in commercially available biota in the Lagos Lagoon, thereby calling for urgent measures to curb microplastic and ultimately plastic pollution. However, there's a need for in-depth research on the effects of environmentally relevant microplastics on edible fish tissues to further elucidate their molecular impact.