Red fermented rice, commonly used as a natural food colorant, contains monacolin K (MK), a bioactive compound known for its cholesterol-lowering properties. However, concerns have emerged regarding its safety due to the potential production of citrinin (CT), a toxic byproduct synthesized during fermentation. To promote safer utilization, this study aimed to determine a fermentation protocol for producing Monascus-biopigment that yields high MK content while minimizing CT concentrations. The study investigated the impact of various parameters over a 10–day fermentation period, including substrate type, initial pH, 0.5% ethanol supplementation, and incubation temperature. Monascus-biopigments were extracted using a 70% (v/v) ethanol solution, and concentrations of both MK and CT were quantified using HPLC-UV-Vis analysis. Among the tested conditions, the control setup, utilizing rice as the fermentation substrate, no pH modification, and incubation at 30°C, resulted in the highest biopigment concentration. Meanwhile, the C3 treatment, which utilized corn as a substrate, a pH adjusted to 2.5, with 0.5% ethanol supplementation, and fermentation temperature of 26°C, yielded the highest MK-to-CT ratio. Although using corn without altering pH enhanced pigment, MK, and CT, lowering the pH to 2.5 significantly suppressed CT concentration. Moreover, the combination of ethanol supplementation and lower temperature further enhanced MK production. In conclusion, the findings provide valuable insights into optimizing fermentation conditions for safer and more potent Monascus-biopigment, supporting its potential application in the food and nutraceutical industries.

Mevinolin (MEV) is an inhibitor of 3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR), which is a crucial enzyme in terpenoid biosynthesis. Since there is limited understanding of the impact of MEV on fruit quality, this study characterized the influence of MEV on metabolism in ‘Shine Muscat’ grapes at the softening stage through an integrated multi-omics analysis. After 7 days of MEV treatment, HMGR and FPPS enzyme activities decreased, accompanied by down-regulation of genes HMGR1, HMGR2, and FPS, while HMGR3 was up-regulated. In the MEP pathway, the activities of key enzymes DXS, TPS, and GGPPS increased, along with upregulation of genes DXR and GGPPS. MEV treatment significantly affected resveratrol and phenolics accumulation, with resveratrol decreasing 3 days post-treatment but elevated after 7 days; however, phenolics were consistently downregulated. Intriguingly, total flavonoids were not significantly affected. Transcriptome analysis showed that MEV upregulated the majority of differentially expressed genes (DEGs), directly affecting ubiquinone and other terpenoid quinone biosynthesis and sesquiterpenoid/triterpenoid biosynthesis pathways. Additionally, MEV treatment also modulated the metabolism of phytohormones, particularly SA, IAA, ETH, and GA. Analysis of volatile metabolites showed that MEV treatment reduced the contents of verbenol and 4-Hexen-1-ol 5-methyl-2-(1-methylethenyl)-,(R)- (Lavandulol), which were related to the cherry and fresh flavors of grapes. Verbenol was positively correlated with Gibberellin 2-beta-dioxygenase8 (GA2ox8), while Lavandulol was negatively correlated with Caffeoyl-CoA O-methyltransferase and beta amylin synthase 1 (β-AS). Specific analysis of terpenoid metabolites revealed that MEV treatment significantly altered 14 terpenoid metabolites, including diterpenoids, sesquiterpenoids, and monoterpenoids, while showing minimal or no effect on triterpenes. Integrated transcriptome analysis demonstrated that isopimaric acid showed strong correlation with the genes alcohol dehydrogenase 1 (ADH1) and geraniol 8-hydroxylase (G8H), with expression patterns significantly altered by MEV treatment. Notably, G8H was negatively correlated with most genes and metabolites. Overall, these findings demonstrate that MEV substantially influences terpenoid biosynthesis and metabolism in “Shine Muscat” grape fruits.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12870-025-07808-9.

Wuyi Hongqu (WYH), also called black-skin-red-koji, which has been utilizing as a fermentation starter for more than one thousand years in China, is a symbiotic combination of Monascus spp. and Aspergillus niger formed through long-term application and domestication. In this study, the strains of Monascus purpureus and A. niger isolated from WYH samples were used to investigate their mutual influence, especially the effects on three main secondary metabolites from M. purpureus, Monascus pigments (MPs), monacolin K (MK), and citrinin (CIT), using a double-sided Petri dish (DSPD). The results showed that co-cultivation of M. purpureus and A. niger strains was favorable to increase the MPs production while inhibiting the CIT production by M. purpureus, especially when M. purpureus strains (M1-1 or M9) were co-cultivated with certain A. niger strains (An1-2 or An9), respectively, and both Monascus strains hardly produced detectable CIT. The expression levels of CIT-related genes in M. purpureus M1-1 or M9 were greatly restricted when co-cultivated with A. niger An1-2 or An9 confirmed by RT-qPCR. This study provides important insights into the selection of WYH production strains and the effects of fungal interactions.

Cytokinins (CKs) are central regulators of leaf senescence, yet their cultivar-specific functions in cereals remain insufficiently understood. Here, we examined dark-induced senescence (DIS) in three barley (Hordeum vulgare L.) cultivars: Carina, Lomerit, and Bursztyn, focusing on responses to exogenous benzyladenine (BA) and inhibition of endogenous CK biosynthesis via the mevalonate (MVA) pathway using lovastatin (LOV). Bursztyn, a winter cultivar, displayed a previously uncharacterized stay-green phenotype, characterized by delayed chlorophyll and protein degradation and reduced sensitivity to BA with respect to chlorophyll retention. In contrast, Carina (spring) senesced rapidly but exhibited strong responsiveness to BA. Lomerit (winter) showed an intermediate phenotype, combining moderate natural resistance to senescence with clear responsiveness to BA. CK application suppressed SAG12 cysteine protease accumulation in all cultivars, serving as a marker of senescence and N remobilization, stabilized photosystem II efficiency, preserved photosynthetic proteins, and alleviated oxidative stress without promoting excessive energy dissipation. Although BA only partially mitigated the decline in net CO2 assimilation, it sustained ribulose-1,5-bisphosphate regeneration, supported electron transport, and stabilized Rubisco and Rubisco activase. Moreover, LOV-based inhibition of the MVA pathway of CK biosynthesis revealed that endogenous CK contributions to senescence delay were most pronounced in Lomerit, moderate in Bursztyn, and negligible in Carina, indicating genotype-specific reliance on MVA-versus methylerythritol phosphate (MEP) pathway-derived CK pools. Collectively, these findings identify Bursztyn as a novel genetic resource for stay-green traits and demonstrate that BA delays DIS primarily by maintaining photosynthetic integrity and redox balance. The results highlight distinct regulatory networks shaping CK-mediated senescence responses in cereals, with implications for improving stress resilience and yield stability.

BackgroundRecent evidence suggests that inhibiting 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR), a key enzyme in cholesterol biosynthesis, has beneficial effects on lipid metabolism and blood pressure (BP), but detrimental consequences on glycemia. Nutraceuticals (NUTs) containing both Monacolin K (MK) and Morus alba have been shown to be more effective in lowering lipids compared to NUT formulations containing only MK. However, the effects of these NUTs on glucose homeostasis have not been fully determined.MethodsTo evaluate the association between LDL-C-lowering therapy and glycemia in patients receiving NUT combinations with or without Morus alba, we analyzed data from a prospective, randomized, active-treatment controlled trial (NCT02898805), which enrolled 359 patients to compare the effects of a NUT combination containing MK alone (Formulation 1, F1; n = 170) versus one containing MK and Morus alba (Formulation 1, F2; n = 189).ResultsParticipants in the two treatment arms (F1 vs. F2) were comparable in terms of sex, age, metabolic parameters, and BP. After 3 months, both groups experienced significant reductions in LDL-C, fasting plasma glucose, HbA1c, and HOMA index. F2 treatment led to a significantly greater reduction in glycemic levels compared to F1 treatment (b = − 16, p < 0.001). Notably, a divergent trend emerged over time: an inverse relationship between LDL-C and glycemic levels was observed in the F1 group, while a significant direct association between LDL-C and glycemic levels was detected in the F2 group (b = 0.06, p = 0.002).ConclusionsTaken together, our findings indicate that the treatment with a NUT combination containing Morus alba simultaneously reduces plasma levels of LDL-C and glucose.

IntroductionRed yeast rice (RYR) is produced through solid-state fermentation by Monascus genus. Its functional component, Monacolin K (MK), has the same structure as lovastatin and can effectively inhibit HMG-CoA reductase, thereby reducing serum cholesterol.MethodsA combinatorial mutagenesis strategy integrating atmospheric room-temperature plasma and heavy-ion radiation was employed to generate mutant strains. The optimizations of substrate components and conditions were carried out during solid-state fermentation (SSF). Subsequently, RYR enriched with MK was produced through scale-up experiments. Additionally, integrated biosynthetic pathway with expression dynamics of MK biosynthetic gene cluster to reveal the efficient biosynthesis of MK in a mutant strain.ResultsA mutant M. purpureus CSUFT-1, which exhibited a remarkable 1.67-fold increase in MK production during SSF compared to the original strain, was obtained. Through refinement of SSF parameters, specifically, adding optimized corn steep liquor adjuvant solution [60% (v/v) corn steep liquor, pH modulation at 5, 3 g/L of MgSO4·7H2O, 1.5 g/L of KH2PO4] with 20% (v/w) of injection volume, the MK yield was significantly amplified to 32.71 mg/g on day 28. Scale-up experiments confirmed the robustness of the optimized process, with MK production stabilizing approximately at 24.66 mg/g. RT-qPCR results showed that seven key genes, including mokC (6.8-fold upregulation) and mokF (22-fold upregulation), were significantly activated during the early stage of fermentation to drive precursor synthesis, only mokE gene exhibited the sustained overexpression in M. purpureus CSUFT-1 during the entire SSF.DiscussionThis work successfully demonstrates a synergistic approach combining advanced combinatorial mutagenesis with precise bioprocess optimization to substantially improve MK yield. The overproducing M. purpureus CSUFT-1 and the optimized SSF protocol provide both microbial resource and technical protocol for industrial-scale production of RYR with high MK.

(1) Background: This study investigated changes in bioactive components and volatile compounds (VCs) during the production of red millet by comparing two varieties: Miao Xiang glutinous millet (waxy) and Jigu-42 (non-waxy). The samples were solid-state-fermented with "Red Ferment" and evaluated for bioactive components. (2) Methods: Multiple analytical methods-including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA) and orthogonal PLS-DA (OPLS-DA), cluster analysis, and correlation analysis-were employed to systematically compare bioactive components and VCs. (3) Results: Significant varietal differences were observed: (1) Miao Xiang glutinous millet showed higher monacolin K (MK) and fatty acid contents; (2) Jigu-42 contained significantly more polyphenols; (3) linoleic acid dominated the fatty acid profiles of two varieties; and (4) a total of twenty-seven VCs were identified, including six alcohols, four aldehydes, seven ketones (corrected from duplicated count), two aromatic hydrocarbons, three heterocycles, one acid, three furans, and one ether. (4) Conclusions: The two varieties exhibited significant differences in MK, pigment profiles, fatty acid composition, polyphenol content, and volatile-compound profiles. These findings provide scientific guidance for the selection of the appropriate millet varieties in functional food production.

Monascus produces various bioactive compounds, including monacolin K (MK), γ-aminobutyric acid (GABA), and Monascus pigments (MPs). Studies have shown that overexpressing genes within the MK biosynthetic cluster significantly enhances MK production. Additionally, MK synthesis in Monascus is regulated by other genes. Based on previous transcriptomic analyses conducted in our laboratory, a significant positive correlation was identified between the expression level of the GAD gene and MK production in M. pilosus. In this study, the GAD gene from M. pilosus was selected for overexpression, and a series of engineered M. pilosus strains were constructed. Among the 20 PCR-positive transformants obtained, 13 strains exhibited MK production increases of 12.84–52.50% compared to the parental strain, while 17 strains showed GABA production increases of 17.47–134.14%. To elucidate the molecular mechanisms underlying the enhanced production of MK and GABA, multi-omics analyses were performed. The results indicated that GAD overexpression likely promotes MK and GABA synthesis in M. pilosus by regulating key genes (e.g., HPD, HGD, and FAH) and metabolites (e.g., α-D-ribose-1-phosphate, β-alanine) involved in pathways such as tyrosine metabolism, phenylalanine metabolism, the pentose phosphate pathway, propanoate metabolism, and β-alanine metabolism. These findings provide theoretical insights into the regulatory mechanisms of MK and GABA biosynthesis in Monascus and suggest potential strategies for enhancing their production.

Triple-negative breast cancer (TNBC), characterized by the absence of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor type 2 (HER2) expression, represents a therapeutic challenge due to its aggressive nature and limited treatment options. Here, we identified the cholesterol-lowering drug lovastatin (LV) as a potent apoptosis-inducing agent in TNBC. Mechanistically, LV disrupts the interaction between the deubiquitinating enzyme USP14 and Survivin, a key anti-apoptotic protein, enhancing polyubiquitination and the proteasomal degradation of Survivin. The overexpression of USP14 was found to stabilize Survivin and rescue LV-induced apoptosis and tumor suppression in vitro and in vivo, whereas USP14 silencing or inhibition with IU1 (a USP14-specific inhibitor) enhanced Survivin turnover and synergized with LV to suppress colony formation in TNBC cells. Clinical relevance was demonstrated through bioinformatic analysis and immunohistochemistry, revealing that elevated Survivin expression in TNBC tissues correlated with poor prognosis and is significantly upregulated in TNBC versus non-TNBC tissues. Our findings identify the USP14–Survivin axis as a potential therapeutic target and highlight LV as a promising candidate for TNBC treatment.

Objective: Neuropathic pain (NP) arises from trauma to the somatosensory nervous system and can be managed using selective serotonin reuptake inhibitors, such as fluoxetine (FLX) and phosphodiesterase inhibitors, such as sildenafil (SD), and cholesterol-lowering agents such as lovastatin (LOVA). The present study aimed to develop and validate an analytical method for the simultaneous estimation of these drugs (SD, FLX, and LOVA [SFL]) using reverse-phase high-performance liquid chromatography (RP-HPLC). Methods: An RP-HPLC method was developed and validated for the quantification of SFL. Chromatographic separation was achieved using a C-18 reverse-phase ODS column with a mobile phase consisting of acetonitrile and 0.2 M ammonium acetate buffer (55:45) in gradient elution mode. The flow rate was maintained at 1 mL/min, and detection was carried out at 228 nm. The method was validated following the ICH Q2 (R2) guidelines, assessing parameters such as linearity, accuracy, precision, limit of detection (LOD), and limit of quantification (LOQ). Results: The developed method exhibited linearity within the concentration range of 20–100 μg/mL, with a regression coefficient (r2) of 0.9992. Retention times for FLX, SD, and LOVA were recorded at 6.481, 4.238, and 19.778 min, respectively. Recovery studies demonstrated an accuracy range of 94.61–110.44%, with a relative standard deviation of 0.06–2.00%, confirming the precision of the method. The LOD values for FLX, SD, and LOVA were found to be 12.77 μg/mL, 14.81 μg/mL, and 13.28 μg/mL, respectively, while the LOQ values were 45.16 μg/mL, 42.33 μg/mL, and 38.71 μg/mL. Conclusion: The validated RP-HPLC method met all required validation criteria and demonstrated suitability for the accurate quantification of FLX, SD, and LOVA in pharmaceutical formulations. These findings support the potential use of these drugs as an alternative therapeutic strategy for NP.

Abstract Introduction: The binding of Programmed Death-Ligand 1 (PD-L1) on the surface of tumor cells to PD-1 on T cells prevents T-cell-mediated anti-cancer immune responses. Immune checkpoint blockades that target the PD-L1/PD-1 interaction have improved survival rates but have several critical limitations. Post-translational modifications (PTMs) of PD-L1, such as glycosylation, are crucial for its stability and engagement with PD-1. Thus, targeting PD-L1 PTMs may be a novel method to improve immune checkpoint blockade efficacy. The mevalonate pathway (MVP) generates various substances involved in PTMs process. Here, we explore the effect of MVP in altering the PTMs of PD-L1 and its impacts on cell surface PD-L1 function. Methods: The triple-negative human breast cancer (TNBC) cell lines MDA-MB-231 and BT549 were treated with the HMG-CoA reductase inhibitor lovastatin (LOV) with or without the addition of IFN-γ and/or MVP intermediates for 48 hours. PD-L1 transcription was evaluated by qPCR. Alterations in PD-L1 size were detected by Western blotting. PD-L1 surface levels were measured by flow cytometry. HMG-CoA reductase (HMGCR) knockdown was performed to validate the non-off-target effect of LOV. The binding of PD-1 protein to the cell surface PD-L1 was measured by PD-1/PD-L1 binding assay. PD-L1 glycosylation levels were measured by Concanavalin A (Con A) lectin binding assay with immunoprecipitated PD-L1 protein. Results: LOV treatment caused a significant molecular weight shift of PD-L1 from approximately 45 kDa to 40 kDa and reduced the cell surface PD-L1 levels. However, PD-L1 transcription was not affected by LOV. HMGCR knockdown produced similar changes, indicating LOV's impact on PD-L1 was not due to off-target effects. The addition of MVP intermediates mevalonate (MEV) or geranylgeraniol, but not farnesol prevented LOV-induced changes in PD-L1 molecular weight and surface levels. Interestingly, the addition of 25-hydroxycholesterol, but not dolichol, partially prevented LOV-induced decreases in PD-L1 surface levels. Geranylgeranyl transferase inhibitor GGTI-286, which prevents protein geranylgeranylation, also altered PD-L1 molecular weight and decreased surface levels. In addition, LOV impaired the binding of PD-1 protein to tumor cells' surface PD-L1, which was reversed by the addition of MVP substrate MEV. Con A lectin-binding of immunoprecipitated PD-L1 showed that the LOV did not reduce the N-glycosylation level of PD-L1. Conclusions: This study reveals the involvement of the MVP in regulating PD-L1 PTMs and highlights the MVP as a potential therapeutic target to modulate PD-L1’s molecular weight and function and improve the efficacy of immunotherapy interventions, presenting a promising avenue for further investigation. Citation Format: Ruoheng Zhang, Emily J. Koubek, Jessie L. Reed, Jeffrey D. Neighbors, Joel M. Reid, Raymond J. Hohl. The role of the mevalonate pathway in post-translational modifications of PD-L1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4265.

Objectives: This research aimed to develop core and coat tablets to enhance the therapeutic efficacy of antilipidemic drugs for treating high cholesterol and triglyceride levels in the blood. Methods: The core and coat tablets were formulated using a combination of two antilipidemic drugs: Lovastatin (LV) and oleanolic acid (OA). LV was incorporated into an immediate-release (IR) layer with various superdisintegrants, while OA formulated into extended-release layer with hydroxypropyl methylcellulose K100. Results: The core and coat tablets were evaluated for the release profiles of both layers, and excipients were optimized. The IR layer of LV achieved complete release within 60 min, while the release of OA was sustained for up to 12 h. Among the formulations tested, LV9 (95.23%) for immediate release, and OA1 (97.13%) for sustained release, were found to be most suitable when scaled at the desired drug release up to 30 min and 12 h, respectively. Stability studies demonstrated that the optimized formulation remained stable without any degradation for 6 months. Pharmacokinetic and pharmacodynamic studies conducted in rabbit models examined the effects of LV/OA tablets on lipid profiles and body weight. Obesity was induced in the rabbits through a high-fat diet. Conclusion: The core and coat LV/OA tablets demonstrated significant efficacy in reducing lipid levels and mitigating weight gain compared to the control group.

Glycine betaine (GB) serves as both a methyl donor and osmoprotectant in microorganisms, facilitating growth and enhancing metabolic product yields. While the polyketide metabolites from Monascus purpureus, such as Monascus pigments (MPs) and monacolin K (MK), have been extensively studied, the effects of GB on their production and the underlying molecular mechanisms remain insufficiently explored. In this study, various concentrations of GB were added to Monascus purpureus M1 cultures, followed by RNA sequencing, RT-qPCR, differential gene expression analysis, and functional enrichment to investigate the regulatory impact of GB on polyketide metabolism. Protein-protein interaction network analysis identified key upregulated genes, including RPS15, RPS14, RPS5, NDK1, EGD2, and ATP9, particularly during the later growth phases. GB significantly upregulated genes involved in stress response, secondary metabolism, and polyketide biosynthesis. Scanning electron microscopy, HPLC, and UV-Vis spectrophotometry further confirmed that GB promoted both strain growth and polyketide production, with red pigment and MK production increasing by 120.08% and 93.4%, respectively. These results indicate that GB enhances growth and polyketide metabolism in Monascus purpureus by functioning as both a methyl donor and osmoprotectant, offering new insights into optimizing microbial polyketide production and revealing gene regulatory mechanisms by GB in Monascus purpureus.

Monacolin K (MK), a secondary metabolite produced by Monascus spp. with the ability to inhibit cholesterol production, is structurally identical to lovastatin produced by Aspergillus terreus. In the lovastatin biosynthetic pathway, the polyketide synthase (PKS) encoded by lovB must work together with the enoyl reductase encoded by lovC to ensure lovastatin production. However, it is unclear whether mokA and mokE in the MK gene cluster of Monascus spp., both of which are highly homologous to lovB and lovC, respectively, also have the same functions for MK biosynthesis. In the current study, the high-yielding MK M. pilosus MS-1 was used as the research object, and it was found that the enoyl reductase domain of MokA may be non-functional due to the lack of amino acids at active sites, a function that may be compensated for by MokE in the MK synthesis pathway. Then, the mokE-deleted (ΔmokE), -complemented (ΔmokE::mokE), and -overexpressed (PgpdA-mokE) strains were constructed, and the results showed that ΔmokE did not produce MK, and ΔmokE::mokE restored MK synthesis, while the ability of PgpdA-mokE to produce MK was increased by 32.1% compared with the original strain MS-1. These results suggest that the MokA synthesized by Monascus spp. must be assisted by MokE to produce MK, just as lovastatin produced by A. terreus, which provides clues for further genetic engineering to improve the yield of MK in Monascus spp.

Tuberculosis (TB) triggers a robust immune response, which leads to significant destruction of the lung tissue at the site of infection, aiding in the transmission of Mycobacterium tuberculosis (Mtb) to the hosts. The excessive inflammatory response contributes heavily to extracellular matrix (ECM) damage, which is linked to high mortality rates among TB patients. Matrix metalloproteinases (MMPs), particularly MMP-2 and MMP-9, are pivotal in the breakdown of the ECM, worsening tissue destruction. In the context of host-directed therapy (HDT), a strategy aimed at modulating the immune response rather than directly targeting the pathogen, we evaluated the potential of lovastatin (LOV). LOV has shown promise in reducing MMP activity and inflammation, which could alleviate the immune-mediated pathology in TB. However, its clinical use has been limited due to poor solubility and biocompatibility, reducing its therapeutic efficacy. To overcome these limitations, we designed inhalable gelatin microspheres (GA-MS) loaded with LOV using the spray-drying technology. This approach improved the solubility and allowed for the controlled release of the drug. The resulting LOV-loaded gelatin microspheres (LOV/GA-MS) had an optimal particle size of 2.395 ± 0.67 μm, facilitating macrophage uptake due to their aerodynamic properties. In in vitro studies using Mtb-infected macrophages, LOV/GA-MS effectively suppressed MMP expression and reduced levels of pro-inflammatory cytokines at a concentration of 20 μg/mL, demonstrating substantial anti-inflammatory potential. Moreover, these microspheres showed a synergistic effect when combined with standard anti-TB drugs, enhancing the overall therapeutic efficacy in in vitro experiments. The findings suggest that inhalable LOV/GA-MS microspheres represent a promising adjunctive host-directed therapy for TB. By modulating the host's immune response and targeting key inflammatory mediators such as MMPs, this approach could mitigate lung tissue damage, improve clinical outcomes, and provide a more holistic treatment option for TB.

In the modern era, rapid high-performance liquid chromatography (HPLC) is the most effective system for the separation of active pharmaceutical ingredients for pharmaceutical analysis due to its extremely effective separation with high sensitivity and accuracy. Establishing and validating an analytical technique using HPLC is essential for upholding the quality of products, as it gives data on accuracy, linearity, precision, and detection. This study focused on developing and validating an HPLC technique for effective evaluation of lovastatin (LV) and oleanolic acid (OA) in core and coat antilipidemic tablets formulated. RP-HPLC process for concurrent assessment of lovastatin also oleanolic acid was performed by means of a Jasco HPLC scheme in addition SunQSil C18 4.6 × 250 mm column (5 μm). The mobile phase utilized for the proposed method was Methanol as well as Water (90:10 v/v), the mobile phase was adjusted to pH 3 using orthophosphoric acid, with a flow rate of 1 mL per min, also finding of both drugs was performed at 210 nm. The validation of the proposed method was carried out in accordance with ICH Q2 (R1) guidelines. Retention time of Lovastatin, in addition to Oleanolic acid, was identified as 4.41 also 6.43 min, respectively. Linearity for Lovastatin also Oleanolic acid was measured in the interval of 2-12 µg/mL with mean percent recovery of 99.99% and 99.82%, respectively. The method demonstrated satisfactory precision, as the percentage relative standard deviation values for both intra- and inter-day precision were below 2%. The proposed simultaneous estimation technique presented exceptional precision, accuracy, linearity, as well as robustness. This simple technique may effectively be applied for the simultaneous estimation of LV as well as OA in therapeutic dosage form. This contributes significantly to ensuring the safety and efficacy of antilipidemic drugs, supporting broader public health goals.

Lovastin (LOV) là một trong những thuốc hàng đầu được lựa chọn trong điều trị mỡ máu. Tuy nhiên, LOV có độ tan kém và thời gian bán hủy ngắn. Vật liệu tổ hợp chitosan/gelatin mang lovastatin (CGL) thường được chế tạo bằng phương pháp dung dịch, các tính chất của vật liệu tổ hợp được đánh giá bằng các phương pháp hiện đại: Phương pháp quang phổ hồng ngoại biến đổi Fourier (FTIR); Phương pháp nhiệt lượng quét vi sai (DSC); Phương pháp hiển vi điện tử quét trường phát xạ (FESEM). Hàm lượng LOV giải phóng từ hệ tổ hợp CGL trong dung dịch đệm pH 2,0 và pH 7,4 có thể được xác định dựa trên độ hấp thụ quang học của dung dịch chứa thuốc bằng phổ tử ngoại - khả kiến (UV-Vis) và các phương trình đường chuẩn của LOV. Nghiên cứu tập trung chế tạo hệ tổ hợp chitosan/gelatin mang LOV với mục đích làm giảm kích thước hạt LOV, đánh giá khả năng kiểm soát và giải phóng LOV ra khỏi hệ tổ hợp chitosan/gelatin. Kết quả phân tích cho thấy các thành phần trong hệ tương tác với nhau bằng tương tác vật lý (liên kết hydro, tương tác lưỡng cực) do vậy, không làm thay đổi cấu tạo hóa học của LOV. Cấu trúc vật lý của LOV chuyển từ dạng tinh thể hình thanh sang dạng hình cầu. LOV giải phóng khỏi hệ tổ hợp CGL trong dung dịch đệm pH 2,0 và pH 7,4 cao hơn so với LOV không được mang bởi tổ hợp chitosan/gelatin.

The increasing use of red yeast rice (RYR) as a natural supplement to manage blood cholesterol levels is driven by its active compound, monacolin K (MK), which is chemically identical to the statin drug lovastatin (LOV). Despite its growing popularity, concerns persists regarding the safety and efficacy of RYR compared to pure statins. This study aimed to evaluate the phytochemical composition, pharmacological effects, and safety profile of various RYR samples in comparison with LOV. RYR samples with different MK content were analyzed using HPLC-DAD to quantify monacolins and other bioactive compounds. The inhibitory activity on HMG-CoA reductase was assessed through an enzymatic assay, while pharmacokinetic properties were predicted using invitro simulated digestion and in silico models. Invitro cytotoxicity was evaluated in intestinal, hepatic, renal, and skeletal muscle cell models. Additionally, the transcriptional levels of muscle damage-related target genes were evaluated by qRT-PCR in skeletal muscle cells treated with a selection of RYR samples. Significant variability in the phytochemical composition of RYR sampleswas observed, particularly in the content of secondary monacolins, triterpenes, and polyphenols. The RYR phytocomplex exhibited superior inhibition of HMG-CoA reductase activity compared to isolated LOV, suggesting synergistic effects between secondary monacolins and other compounds. Molecular insights revealed that RYR samples had a lower impact on muscle cells than LOV, as reflected also by cell viability. These findings suggest that RYR could serve as a safe alternative to purified statins. However, further research is needed to fully elucidate the mechanisms behind the synergistic activity of the phytocomplex and to firmly establish the clinical efficacy of this natural product.

Monascus species are capable of producing various active metabolites, including monacolin K (MK) and pigments. Studies have shown that the overexpression of the mok I gene from the MK synthesis gene cluster in Monascus species can significantly increase MK production; however, the molecular mechanism has not yet been fully elucidated. Therefore, this study focused on the mok I gene of Monascus pilosus to construct overexpression strains of the mok I gene, resulting in high-yield MK production. Sixteen positive transformants were obtained, seven of which produced 9.63% to 41.39% more MK than the original strain, with no citrinin detected in any of the transformants. The qRT-PCR results revealed that the expression levels of mok I in the transformed strains TI-13, TI-24, and TI-25 increased by more than 50% compared to the original strain at various fermentation times, with the highest increase being 10.9-fold. Furthermore, multi-omics techniques were used to analyze the molecular mechanisms underlying enhanced MK production in transformed strains. The results indicated that mok I overexpression may enhance MK synthesis in M. pilosus by regulating the expression of key genes (such as MAO, HPD, ACX, and PLC) and the synthesis levels of key metabolites (such as delta-tocopherol and alpha-linolenic acid) in pathways linked to the biosynthesis of cofactors, the biosynthesis of unsaturated fatty acids, tyrosine metabolism, ubiquinone and other terpenoid-quinone biosynthesis, alpha-linolenic acid metabolism, and glycerophospholipid metabolism. These findings provide a theoretical basis for further study of the metabolic regulation of MK in Monascus species and for effectively enhancing their MK production.

Monacolin K (MK), produced by Monascus fermentation, has significant hypolipidemic effects. In this study, under solid-state fermentation (SSF) quinoa was used as substrate to investigate the effect of the MK production by Monascus species quinoa. The results of the single factor experiment showed that the amount of the substrate loaded, addition of water to the substrate, and fermentation temperature change time have a significant effect on the MK production of Monascus-fermented quinoa. It was found that neither M. pilosus-fermented quinoa nor M. ruber-fermented quinoa produced citrinin. Consequently, these three factors were selected for the response surface optimization to determine the optimal fermentation process. The results showed that the optimal fermentation process of M. pilosus-fermented quinoa was as follows: quinoa loading amount of 30 g/300 ml, addition of 20 ml water to substrate, seed liquid inoculation volume of 10%, cultured at 30°C for 2 days, followed by a temperature change to 25°C and continued cultivation until 12 days. Under this fermentation process, the yield of MK reached 2.51 mg/g and without production of any citrinin. The optimal fermentation process of M. ruber-fermented quinoa was as follows: quinoa loading amount of 31 g/300 ml, water addition of substrate of 20 ml, seed liquid inoculation volume of 20%, cultured at 30°C for 2 days, followed by a temperature change to 25°C and continued cultivation until 12 days. Under this fermentation process, the yield of MK reached 3.22 mg/g and without any citrinin production. Bangladesh J. Bot. 53(3): 717-729, 2024 (September) Special