Showing papers on "Lovastatin published in 2018"
TL;DR: This study provides an alternative method for production of monacolin J and lovastatin and reveals the potential of a methylotrophic yeast to produce complicated pharmaceuticals from methanol.
98 citations
TL;DR: In this article, the response surface methodology (RSM) was applied to optimize four different fermentation parameters: temperature (25−35 °C), time (10−20 days), glucose (0.1−1.5%), and peptone concentration.
Abstract: In this study, Saccharina japonica (brown seaweed) was fermented in solid state by Monascus purpureus to maximize lovastatin production, the response surface methodology (RSM) was applied to optimize four different fermentation parameters: temperature (25−35 °C), time (10−20 days), glucose (0.1–1.5%) and peptone (0.1–0.7%) concentration. The predicted combination of process parameters yielding the highest rate of lovastatin production (13.40 mg gdfs−1) was obtained at a temperature of 25.64 °C, a fermentation time of 14.49 days, glucose concentration of 1.32% and peptone concentration of 0.20%, with 92.85% validity. Among the studied factors, glucose, incubation time and temperature most strongly influenced lovastatin production. Triplicate experiments in which lovastatin was produced under optimal conditions resulted in a mean yield of 13.98 mg gdfs−1 with 207.84 mg gdfs−1 biomass. Liquid chromatography–tandem mass spectrometry (quadrupole time-of-flight) analysis confirmed the ionic molecular weight of lovastatin (405.26). Lovastatin from the M. purpureus fermented S. japonica exhibited thermal stability, superoxide dismutase activity and a cholesterol esterase inhibition activity that was higher than that of unfermented sample and showed no toxic effect on Caco-2 cell. Thus, S. japonica was a suitable substrate to maximize lovastatin production from M. purpureus at optimum condition for applying in food and pharmaceutical industry.
34 citations
TL;DR: Combination of curcumin and berberine exhibited better ameliorative effects in treating NAFLD than lovastatin, and this enhanced effect is associated with oxidative stress, hepatic inflammation and lipid metabolism.
32 citations
TL;DR: The study confirms the efficacy of the pathogenesis‐targeted therapy and introduces the possibility of modifying its formula by adding glycolic acid in order to improve the treatment.
Abstract: Congenital Hemidysplasia with Ichthyosiform nevus and Limb Defects (CHILD syndrome) is a rare X-linked dominant genodermatosis caused by mutations in the NAD(P) dependent steroid dehydrogenase-like protein gene. Its defect leads to accumulation of toxic metabolic intermediates upstream from the pathway block and to the deficiency of bulk cholesterol, probably leading to altered keratinocyte membrane function, resulting in the phenotype seen in CHILD syndrome. Symptomatic treatment using emollients and retinoids to reduce scaling has long been used until recently, whereby new therapeutic means based on the pathogenesis-targeted therapy have been developed. We subsequently chose to use the same pathogenesis-based therapy using a 2% cholesterol and 2% lovastatin cream with or without glycolic acid in two of our patients. Improvement in CHILD skin lesions was seen as early as 4 weeks after initiation. The addition of glycolic acid helped improve the penetrance of the cholesterol and lovastatin cream into the thick waxy scales. Our study confirms the efficacy of the pathogenesis-targeted therapy and introduces the possibility of modifying its formula by adding glycolic acid in order to improve the treatment.
26 citations
TL;DR: It was concluded that the lovastatin and simvastatin efficiently ameliorated the OVX-induced osteoporosis and showed more resemblance to the normal group in terms of BMD, expression of osteogenic genes, serum biochemical parameters, histomorphometric findings, and biomechanical performance with no significant side-effects.
Abstract: In addition to lipid-lowering properties, statins have been suggested to affect bone turnover by increasing the osteoblastic bone formation and blocking the osteoclastogenesis. However, there are many controversial reports regarding the beneficial effect of statins on osteoporosis. In this study, we investigated the therapeutic effects of the most important lipophilic statins administered orally for 60 days to the ovariectomized (OVX) female Sprague–Dawley rats and compared the effects on different harvested trabecular and compact bones. Thirty female rats were divided into five equal groups including the normal rats, untreated OVX rats (negative control), and the OVX rats treated with atorvastatin (20 mg/kg/day), simvastatin (25 mg/kg/day), and lovastatin (20 mg/kg/day). The osteoporotic animals were treated daily for 60 days and euthanized at the end of experiments. The effectiveness of these treatments was evaluated by biomechanical testing, histopathologic, histomorphometric, micro-CT scan, real-time PCR, and serum biochemical analysis. Moreover, the hepatotoxicity and rhabdomyolysis related with these treatments were assessed by biochemistry analysis and histopathological evaluation. The results and statistical analysis showed that systemic delivery of simvastatin and lovastatin significantly increased serum calcium level, expression of osteogenic genes, bone mineral density (BMD), and biomechanical properties in comparison to the untreated OVX rats, especially in trabecular bones (P < 0.05). The results of different analysis also indicated that there was no statistical difference between the atorvastatin-treated animals and the negative control. Among all treatments, only atorvastatin showed an evident hepatotoxicity and myopathy. It was concluded that the lovastatin and simvastatin efficiently ameliorated the OVX-induced osteoporosis. Moreover, the simvastatin-treated animals showed more resemblance to the normal group in terms of BMD, expression of osteogenic genes, serum biochemical parameters, histomorphometric findings, and biomechanical performance with no significant side-effects.
24 citations
TL;DR: The possibility that lovastatin can be useful for treating IBS is indicated by the possibility that Lovastatin improves symptoms in the patients with IBS by attenuating these visceral changes.
22 citations
TL;DR: Liposomal formulation obtained combining DOPC, DOPE and hydro-alcoholic extract of RYR showed optimal physico-chemical, mechanical and thermal characteristics and the strongest inhibition activity versus 3-hydroxy-3-Methyl glutaryl coenzyme A (HMG-CoA) reductase.
22 citations
TL;DR: Inhibition of Rac1 signaling is of major relevance for the cardioprotective effects of lovastatin in the context of anthracycline-induced cardiotoxicity and EAT is a useful marker of subacute cardiot toxicity caused by persisting harmful effects of doxorubicin.
Abstract: The anticancer efficacy of anthracyclines is limited by congestive heart failure. Clinically established markers of early onset of cardiotoxicity following anthracycline treatment and preventive measures are missing. Although statins are reported to alleviate anthracycline-induced cardiotoxicity in vivo, the molecular mechanisms involved remain elusive. In vitro data point to Rac1 as major target of the cytoprotective statin effects. Here we investigated whether specific inhibition of Rac1 by NSC23766 is as effective as lovastatin in preventing subacute cardiotoxicity following doxorubicin treatment. C57BL/6 mice were treated over 3 weeks with multiple low doses of doxorubicin (6 × 3 mg/kg BW, i.p.) and the level of DNA damage, apoptosis and regenerative proliferation as well as pro-inflammatory, pro-fibrotic and oxidative stress responses were investigated. Moreover, heart function was monitored by echocardiography. Doxorubicin induced subacute cardiotoxicity which was reflected on the level of residual DNA damage, frequency of apoptotic and mitotic cells as well as elevated mRNA expression of markers of heart failure, remodeling and mitochondrial biogenesis. These molecular markers of cardiotoxicity were mitigated to a similar extent by co-treatment with either lovastatin (10 mg/kg BW, p.o.) or NSC23766 (5 mg/kg BW, i.p.) three times a week. Moreover, doxorubicin caused diastolic dysfunction as reflected by increased E-wave acceleration time (EAT), which again was prevented by pharmacological inhibition of Rac1. Inhibition of Rac1 signaling is of major relevance for the cardioprotective effects of lovastatin in the context of anthracycline-induced cardiotoxicity. Moreover, EAT is a useful marker of subacute cardiotoxicity caused by persisting harmful effects of doxorubicin.
21 citations
TL;DR: A shift in rumen fermentation pathway to favor propionate production which serves as H+ sink, partly explaining the observed CH4 reduction.
Abstract: Twenty male Saanen goats were randomly assigned to four levels of lovastatin supplementation and used to determine the optimal dosage and sustainability of naturally produced lovastatin from fermentation of palm kernel cake (PKC) with Aspergillus terreus on enteric methane (CH4) mitigation. The effects on ruminal microbiota, rumen fermentation, feed digestibility and health of animal were determined over three measuring periods (4-, 8- and 12-weeks) and the accumulation of lovastatin in tissues was determined at the end of the experiment. The diets contained 50% rice straw, 22.8% concentrates and 27.2% of various proportions of untreated or treated PKC to achieve the target daily intake level of 0 (Control), 2, 4 or 6 mg lovastatin/kg body weight (BW). Enteric CH4 emissions per dry matter intake (DMI), decreased significantly (P<0.05) and equivalent to 11% and 20.4%, respectively, for the 2 and 4 mg/kg BW groups as compared to the Control. No further decrease in CH4 emission thereafter with higher lovastatin supplementation. Lovastatin had no effect on feed digestibility and minor effect on rumen microbiota, and specifically did not reduce the populations of total methanogens and Methanobacteriales (responsible for CH4 production). Similarly, lovastatin had little effect on rumen fermentation characteristics except that the proportion of propionate increased, which led to a decreasing trend (P<0.08) in acetic: propionate ratio with increasing dosage of lovastatin. This suggests a shift in rumen fermentation pathway to favor propionate production which serves as H+ sink, partly explaining the observed CH4 reduction. No adverse physiological effects were noted in the animals except that treated PKC (containing lovastatin) was less palatable at the highest inclusion level. Lovastatin residues were detected in tissues of goats fed 6 mg lovastatin/kg BW at between 0.01 to 0.03 μg/g, which are very low.
21 citations
TL;DR: An oral dosage form containing lovastatin and coated with 2 different enteric polymers that enabled a pH-dependent "dual pulse" drug release is developed and shows promise in treating IBS-C.
19 citations
TL;DR: By comparing protein expression in the cortical white matter tracts from mice at different postnatal days, it is identified that in the NPC1 mutant (NPC1-/-) mice, the onset of myelination is delayed and the amount of the major myelin protein MBP and PLP, and oligodendrocyte regulatory factor Olig1 and Olig2, but not NG2 and Sox10, decreased significantly, suggesting a disruption of oligodendedrocytes differentiation.
TL;DR: The findings provide guidelines for the co‐administration of sinomenine with simvastatin or lovastatin in clinic and show that the long‐term administration of simVastatin could up‐regulate the expression of CYP3A1/2 to account for the effect.
TL;DR: The results demonstrated that lovastatin in the treated rice straw acted specifically on the methanogens by inhibiting the activity of HMG-CoA reductase in the meethanogens’ cell membrane biosynthesis pathway and thus the growth of rumen methanogen as previously reported.
Abstract: The objectives of this study were to test the efficacy of producing lovastatin in rice straw treated with Aspergillus terreus in larger laboratory scale following the procedure previously reported and to investigate the effectiveness of the treated rice straw containing lovastatin on methane mitigation in goats. The concentration of lovastatin in the treated rice straw was 0.69 ± 0.05 g/kg dry matter (DM) rice straw. Our results showed that supplementation of lovastatin at 4.14 mg/kg BW reduced methane production by 32% while improving the DM digestibility by 13% (P < 0.05) in goats fed fermented rice straw compared to those fed untreated rice straw. Populations of total methanogens and Methanobacteriales species were significantly reduced (P < 0.05) while the population of total bacteria and Ruminococcus albus were increased in the treatment group (P < 0.05). Our results demonstrated that lovastatin in the treated rice straw acted specifically on the methanogens by inhibiting the activity of HMG-CoA reductase in the methanogens’ cell membrane biosynthesis pathway and thus the growth of rumen methanogens as previously reported. This study provides a simple yet practical approach to mitigate enteric methane production particularly in the developing countries which depend heavily on the use of agro-biomass such as rice straw to feed their ruminant animals.
TL;DR: Oral medication with a combination of lovastatin and AICAR, which are individually known to have immunomodulatory effects, provides potent protection and repair of inflammation‐induced loss and dysfunction of mitochondria and peroxisomes as well as myelin and axonal abnormalities in EAE.
Abstract: Recent studies report that loss and dysfunction of mitochondria and peroxisomes contribute to the myelin and axonal damage in multiple sclerosis (MS). In this study, we investigated the efficacy of a combination of lovastatin and AMP-activated protein kinase (AMPK) activator (AICAR) on the loss and dysfunction of mitochondria and peroxisomes and myelin and axonal damage in spinal cords, relative to the clinical disease symptoms, using a mouse model of experimental autoimmune encephalomyelitis (EAE, a model for MS). We observed that lovastatin and AICAR treatments individually provided partial protection of mitochondria/peroxisomes and myelin/axons, and therefore partial attenuation of clinical disease in EAE mice. However, treatment of EAE mice with the lovastatin and AICAR combination provided greater protection of mitochondria/peroxisomes and myelin/axons, and greater improvement in clinical disease compared with individual drug treatments. In spinal cords of EAE mice, lovastatin-mediated inhibition of RhoA and AICAR-mediated activation of AMPK cooperatively enhanced the expression of the transcription factors and regulators (e.g. PPARα/β, SIRT-1, NRF-1, and TFAM) required for biogenesis and the functions of mitochondria (e.g. OXPHOS, MnSOD) and peroxisomes (e.g. PMP70 and catalase). In summary, these studies document that oral medication with a combination of lovastatin and AICAR, which are individually known to have immunomodulatory effects, provides potent protection and repair of inflammation-induced loss and dysfunction of mitochondria and peroxisomes as well as myelin and axonal abnormalities in EAE. As statins are known to provide protection in progressive MS (Phase II study), these studies support that supplementation statin treatment with an AMPK activator may provide greater efficacy against MS.
TL;DR: A combinational approach consisting of drugs designed for targeting lipid metabolism, lovastatin and troglitazone, exhibits anti-proliferation in cell culture systems and leads to tumor regression in a mouse xenograft model, supporting a hypothesis that the inhibitory effect of the combination is partly through a cell cycle arrest at G0/G1 phase.
Abstract: Anaplastic thyroid cancer (ATC) is a malignant subtype of thyroid cancers and its mechanism of development remains inconclusive. Importantly, there is no effective strategy for treatment since ATC is not responsive to conventional therapies, including radioactive iodine therapy and thyroid-stimulating hormone suppression. Here, we report that a combinational approach consisting of drugs designed for targeting lipid metabolism, lovastatin (an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, HMGCR) and troglitazone (an agonist of peroxisome proliferator-activated receptor gamma, PPARγ), exhibits anti-proliferation in cell culture systems and leads to tumor regression in a mouse xenograft model. The composition contains a sub-lethal concentration of both drugs and exhibits low toxicity to certain types of normal cells. Our results support a hypothesis that the inhibitory effect of the combination is partly through a cell cycle arrest at G0/G1 phase, as evidenced by the induction of cyclin-dependent kinase inhibitors, p21cip and p27kip, and the reduction of hyperphosphorylated retinoblastoma protein (pp-Rb)-E2F1 signaling. Therefore, targeting two pathways involved in lipid metabolism may provide a new direction for treating ATC.
TL;DR: The whole genome sequencing of A. terreus and a selected endophytic fungus, Diaporthe ampelina (Phomopsis) was performed, and there was a complete absence of lovastatin gene cluster in the genome of D. ampeline (an endophyte).
Abstract: Lovastatin is a competitive inhibitor of the enzyme hydroxymethyl glutaryl coenzyme A reductase (HMGR) in cholesterol biosynthetic pathway and hence used in the treatment of hyperlipidemia. In a previous study, we report a tropical soil isolate, Aspergillus terreus (KM017963), which produces ample amount of lovastatin than its counterpart that are endophytic in origin. Bioinformatic analysis of whole genome sequence of A. terreus (AH007774.1), a soil isolate revealed the presence of gene cluster (AF141924.1 & AF141925.1) responsible for lovastatin production, whereas endophytic fungi including a strain of A. terreus showed no homology with the lovastatin gene cluster. The molecular study was also carried out targeting PCR amplification of the two important genes, lovE (a regulatory gene) and lovF (transcriptional regulatory factor) in genomic and c-DNA of soil and endophytic fungi. Expression of the two genes was successful in A. terreus (KM017963), whereas the same was not achieved in endophytic fungi. To further validate our above findings, in the present study, the whole genome sequencing of A. terreus and a selected endophytic fungus, Diaporthe ampelina (Phomopsis) was performed. Lovastatin gene cluster, when aligned on the consensus sequence of both genomes, the entire lovastatin gene cluster was detected in a single scaffold (1.16) of A.terreus genome. On the contrary, there was a complete absence of lovastatin gene cluster in the genome of D. ampelina (an endophyte). The probable reasons for the absence of lovastatin gene cluster in endophytic fungi are discussed.
TL;DR: In this paper, the best Monascus purpureus strain was selected from MTCC 369, 410, 1090 based on lovastatin concentration, and four medium parameters (NH 4 Cl, MgSO 4, NaCl, CaCl 2 ) screened by Plackett-Burman design from total nine medium variables were optimised by Box-Behnken design of response surface methodology.
Abstract: Angkak (red mold rice, red yeast rice, Chinese red rice), a traditional Chinese functional food is produced by solid-state fermentation of cooked non-glutinous rice with Monascus species. The secondary metabolite of Monascus species, monacolin K (lovastatin), has been proved to lower blood lipid levels. In this study, the best Monascus purpureus strain was selected from MTCC 369, 410, 1090 based on lovastatin concentration. Four medium parameters (NH 4 Cl, MgSO 4 , NaCl, CaCl 2 ) screened by Plackett-Burman design from total nine medium variables were optimised by Box-Behnken design of response surface methodology. Maximum lovastatin production of 3.420 mg/g was predicted in the solid medium containing 20 g rice and 40 ml liquid nutrient (NH 4 Cl 14.32 g/l, MgSO 4 0.76 g/l, NaCl 14.65 g/l, and CaCl 2 0.54 g/l) by the point prediction tool of Design Expert Ver. 7.1 software at 14 th day of fermentation.
TL;DR: Lovastatin treatment decreased CYR61 expression, inhibited cell invasion and altered epithelial-to-mesenchymal-transition (EMT)-related protein expression, while CYR 61 overexpression abolished the effect of lovastatin.
Abstract: Background/aims Preventing cell metastasis is an effective therapeutic strategy to treat osteosarcoma and improve prognosis. Statins have been found to have anticancer effects in addition to their cholesterol-lowering action. As a new target of statins, cysteine-rich 61 (CYR61) was recently identified to promote cell migration and metastasis in osteosarcoma. However, the underlying mechanisms mediating the regulation of CYR61 expression by statins remain unknown. Methods Human osteosarcoma cell lines MG63 and SaOS2 were used to clarify the effect of lovastatin on CYR61 expression. Real-time PCR was performed to detect mRNA or microRNA (miRNA) levels and western blot was performed to detect protein levels. Cell invasive ability was determined using Transwell assays. Lentivirus encoding CYR61 cDNA or sterol regulatory element-binding protein 2 (SREBP-2) shRNA was used to upregulate CYR61 expression or downregulate SREBP-2 expression. Binding of the CYR61 3' untranslated region (UTR) and miR-33a was analyzed by luciferase reporter assay. Results We found that lovastatin treatment decreased CYR61 expression, inhibited cell invasion and altered epithelial-to-mesenchymal-transition (EMT)-related protein expression, while CYR61 overexpression abolished the effect of lovastatin. Moreover, lovastatin increased the expression of SREBP-2 and miR-33a, which were then downregulated by SREBP-2 silencing. Bioinformatics analysis indicated that the CYR61 3'UTR harbored a potential miR-33a binding site and luciferase reporter assay demonstrated that CYR61 was a target of miR-33a in osteosarcoma cells. Furthermore, miR-33a could inhibit cell invasion and alter EMT-related protein expression. SREBP-2 silencing or miR-33a inhibitor upregulated CYR61 expression and reversed the effects of lovastatin on cell invasion and EMT-related proteins. Conclusion Our findings suggest lovastatin suppresses osteosarcoma cell invasion through the SREBP-2/miR-33a/CYR61 pathway.
TL;DR: It is found that while lovastatin normalizes excessive hippocampal protein synthesis and reduces audiogenic seizures in the Fmr1-/y mouse, simvastatin does not correct either phenotype, caution against the assumption that simVastatin is a valid alternative to lovASTatin for the treatment of FX.
Abstract: The cholesterol-lowering drug lovastatin corrects neurological phenotypes in animal models of fragile X syndrome (FX), a commonly identified genetic cause of autism and intellectual disability. The therapeutic efficacy of lovastatin is being tested in clinical trials for FX, however the structurally similar drug simvastatin has been proposed as an alternative due to an increased potency and brain penetrance. Here, we perform a side-by-side comparison of the effects of lovastatin and simvastatin treatment on two core phenotypes in the Fmr1-/y mouse model. We find that while lovastatin normalizes excessive hippocampal protein synthesis and reduces audiogenic seizures (AGS) in the Fmr1-/y mouse, simvastatin does not correct either phenotype. These results caution against the assumption that simvastatin is a valid alternative to lovastatin for the treatment of FX.
TL;DR: A methylotrophic yeast strain producing 419.5 mg/L lovastatin was constructed by optimization of biosynthetic gene dosages and coexpression of a statin pump protein, TapA, which proved that P. pastoris is a promising chassis organism for natural-product biosynthesis.
Abstract: Methanol has attracted interest as a substrate for improvement of product titers and yields of bioprocesses, because methanol has a more reduced state than sugars do and can be renewably synthesized from abundant natural gas supplies. Our aim was to engineer methylotrophic Pichia pastoris to convert methanol into value-added lovastatin more productively. A strengthened biosynthetic pathway of lovastatin was constructed through the assembly of three modules with increasing module-specific antibiotic stress in the methylotrophic yeast P. pastoris. The resulting strain (P. p/LV_V#9) produced 287.5 ± 2.0 mg/L lovastatin in a 5-L bioreactor from methanol. The production was further improved by identification and overexpression of a statin pump protein, TapA, a membrane protein capable of lovastatin efflux out of the cell. A TapA-overexpressing strain, P. p/LV_V#9-TapA, produced 419.0 ± 9.5 mg/L lovastatin from methanol: 46% more than P. p/LV_V#9 did, and 520% more relative to the strain (P. p/LV_SC) with single-copy genes. A methylotrophic yeast strain producing 419.0 ± 9.5 mg/L lovastatin was constructed by optimization of biosynthetic gene dosages and coexpression of a statin pump protein; these results proved that P. pastoris is a promising chassis organism for natural-product biosynthesis. A membrane protein, TapA was found to perform the function of exporting intracellular lovastatin and enhanced lovastatin production.
TL;DR: Peptonized milk, linoleic acid, glucose, yeast extract, and trace elements had a positive impact on the production of lovastatin by A. terreus.
Abstract: The production of lovastatin by Aspergillus terreus PU-PCSIR-1 was studied using submerged fermentation technique. Effects of pH, incubation temperature, incubation time, and the media components on the production of lovastatin were investigated. Maximum lovastatin yield (198.90 mg/L) was achieved after 14 days of incubation at temperature 28°C and pH 7.4. Peptonized milk, linoleic acid, glucose, yeast extract, and trace elements had a positive impact on the production of lovastatin by A. terreus.
TL;DR: A fast approach for the assay of lovastatin in both form, lactone and acid, by mass spectrometry is presented and the accuracy values ranged from 97 to 101%; the analytical parameters values of LOQ, LOD, recovery and reproducibility were calculated analyzing fortified samples, confirming the reliability of the proposed approach.
Abstract: Monacolin K, the active ingredient present in dietary supplement, is a nutraceutical whose health benefits have been widely documented. A fast approach for the assay of lovastatin in both form, lactone and acid, by mass spectrometry is presented. The quantitative assay is carried out by HPLC-MS/MS using the multiple reaction monitoring (MRM) mode and simvastatin and pravastatin as internal standards. The accuracy values ranged from 97 to 101%; the analytical parameters values of LOQ, LOD, recovery and reproducibility, were calculated analyzing fortified samples, confirming the reliability of the proposed approach.
TL;DR: An improved microbial cell factory can realize single-step bioproduction of monacolin J in a more efficient way, providing an attractive and eco-friendly substitute over the existing chemical synthetic routes of monAColin J and promoting complete biopProduction of simvastatin at industrial scale.
Abstract: Biosynthesis of simvastatin, the active pharmaceutical ingredient of cholesterol-lowering drug Zocor, has drawn increasing global attention in recent years. Although single-step in vivo production of monacolin J, the intermediate biosynthetic precursor of simvastatin, has been realized by utilizing lovastatin hydrolase (PcEST) in our previous study, about 5% of residual lovastatin is still a problem for industrial production and quality control. In order to improve conversion efficiency and reduce lovastatin residues, modification of PcEST is carried out through directed evolution and a novel two-step high-throughput screening method. The mutant Q140L shows 18-fold improved whole-cell activity as compared to the wild-type, and one fold enhanced catalytic efficiency and 3 degrees C increased T-50(10) over the wild-type are observed by characterizing the purified protein. Finally, the engineered A. terreus strain overexpressing Q140L mutant exhibited the increased conversion efficiency and the reduced lovastatin residues by comparing with A. terreus strain overexpressing the wild-type PcEST, where almost 100% of the produced lovastatin is hydrolyzed to monacolin J. Therefore, this improved microbial cell factory can realize single-step bioproduction of monacolin J in a more efficient way, providing an attractive and eco-friendly substitute over the existing chemical synthetic routes of monacolin J and promoting complete bioproduction of simvastatin at industrial scale.
TL;DR: The data suggest that ERK1/2-signaling activation in dyskinetic animals is maximal and does not require further genetic enhancement in the upstream Ras pathway, but also demonstrate that such a genetic enhancement may reduce the efficacy of anti-dyskinetic drugs like lovastatin.
Abstract: Increasing evidence supports a close relationship between Ras-ERK1/2 activation in the striatum and L-DOPA-induced dyskinesia (LID). ERK1/2 activation by L-DOPA takes place through the crosstalk between D1R/AC/PKA/DARPP-32 pathway and NMDA/Ras pathway. Compelling genetic and pharmacological evidence indicates that Ras-ERK1/2 inhibition prevents LID onset and may even revert already established dyskinetic symptoms. However, it is currently unclear whether exacerbation of Ras-ERK1/2 activity in the striatum may further aggravate dyskinesia in experimental animal models. Here we took advantage of two genetic models in which Ras-ERK1/2 signaling is hyperactivated, the Nf1+/− mice, in which the Ras inhibitor neurofibromin is reduced, and the Ras-GRF1 overexpressing (Ras-GRF1 OE) transgenic mice in which a specific neuronal activator of Ras is enhanced. Nf1+/− and Ras-GRF1 OE mice were unilaterally lesioned with 6-OHDA and treated with an escalating L-DOPA dosing regimen. In addition, a subset of Nf1+/− hemi-parkinsonian animals was also co-treated with the Ras inhibitor lovastatin. Our results revealed that Nf1+/− and Ras-GRF1 OE mice displayed similar dyskinetic symptoms to their wild-type counterparts. This observation was confirmed by the lack of differences between mutant and wild-type mice in striatal molecular changes associated to LID (i.e., FosB, and pERK1/2 expression). Interestingly, attenuation of Ras activity with lovastatin does not weaken dyskinetic symptoms in Nf1+/− mice. Altogether, these data suggest that ERK1/2-signaling activation in dyskinetic animals is maximal and does not require further genetic enhancement in the upstream Ras pathway. However, our data also demonstrate that such a genetic enhancement may reduce the efficacy of anti-dyskinetic drugs like lovastatin.
03 Aug 2018
TL;DR: Structural analysis including NMR and LC–MS of the purified lovastatin reassures the molecular formula and molecular weight similar to standard, and Lovastatin showed significant antioxidant property with IC50.
Abstract: Beside anti-cholesterol activity, lovastatin garners worldwide attention for therapeutical application against various diseases especially cancer. A total of 36 filamentous fungi from soil samples were isolated and screened for lovastatin production by yeast growth bioassay method. C9 strain (later identified as Cunninghamella blakesleeana) was screened as potential strain of lovastatin production. Further confirmation of the compound was made using TLC, HPTLC and HPLC in which similar Rf value, densitogram peak and chromatogram peak against the standard lovastatin were observed, respectively. The purified lovastatin subjected for IR analysis showed a lactone ring peak at 1763.63 cm−1 similar to standard lovastatin. Further structural analysis including NMR and LC–MS of the purified lovastatin reassures the molecular formula and molecular weight similar to standard. In quantitative terms, C. blakesleeana, Aspergillus terreus and Aspergillus flavus produced 1.4 mg g−1 DWS, 0.83 mg g−1 DWS and 0.3 mg g−1 DWS of lovastatin, respectively, (p < 0.0001) without any optimization. Lovastatin showed significant antioxidant property with IC50: 145.9 µg mL−1 (140 µL), and the percentage of inhibition is maximum at 199.5 µg/mL which is statistically significant (p < 0.0001).
Journal Article•
TL;DR: It was determined that optimized conditions for the hyper-production of lovastatin from fungal sources have a significant effect.
Abstract: Lovastatin is a natural competitive inhibitor of 3-hydroxy-3-methyl glutaryl coenzyme-A (HMG-CoA) reductase and inhibits specifically rate limiting step in cholesterol biosynthesis. Further, lovastatin in comparison with synthetic drugs has no well-reported side effects. Four pure isolated filamentous fungal strains including Aspergillus niger IBL, Aspergillus terreus FFCBP-1053, Aspergillus flavus PML and Aspergillus nidulans FFCBP-014 have been cultured by solid state fermentation (SSF) using rice straw as substrate for the synthesis of lovastatin. After selecting Aspergillus terreus FFCBP-1053 as the best producer of lovastatin, various selected physical parameters including pH, temperature, inoculums size and moisture content were optimized through response surface methodology (RSM) under center composite design (CCD) for lovastatin hyper production. Maximum lovastatin production of 2070±91.5 was predicted by the quadratic model in the medium having moisture content 70% and pH 4.5 at 35°C which was verified experimentally to be 2140±93.25µg/g DW of FM (microgram/gram dry weight of fermentation medium), significantly (P 0.05) measured by spectrophotometer at 245 nm against standard. It was determined that optimized conditions for the hyper-production of lovastatin from fungal sources have a significant effect.
TL;DR: Evidence is provided that isoflavones and Lv in XZK act in concert to enhance the efficacy and reduce the side effects of Lv, for the first time.
Abstract: Xuezhikang capsule (XZK) is a traditional Chinese medicine that contains lovastatin (Lv) for hyperlipidemia treatment, although it has fewer side effects than Lv. However, the pharmacokinetic mechanisms contributing to its distinct efficacy and low side effects are unclear. Mice were fed a high-fat diet (HFD) for 6 weeks to induce hyperlipidemia. We first conducted the pharmacokinetic studies in HFD mice following oral administration of Lv (10 mg/kg, i.g.) and found that HFD remarkably decreased the active form of Lv (the lovastatin acid, LvA) exposure in the circulation system, especially in the targeting organ liver, with a declined conversion from Lv to LvA, whereas the Lv (responsible for myotoxicity) exposure in muscle markedly increased. Then we compared the pharmacokinetic profiles of Lv in HFD mice after the oral administration of XZK (1200 mg/kg, i.g.) or an equivalent dose of Lv (10 mg/kg, i.g.). A higher exposure of LvA and lower exposure of Lv were observed after XZK administration, suggesting a pharmacokinetic interaction of some ingredients in XZK. Further studies revealed that HFD promoted the inflammation and inhibited carboxylesterase (CES) activities in the intestine and the liver, thus contributing to the lower transformation of Lv into LvA. In contrast, XZK inhibited the inflammation and upregulated CES in the intestine and the liver. Finally, we evaluated the effects of monacolins and phytosterols, the fractional extracts of isoflavones, on inflammatory LS174T or HepG2 cells, which showed that isoflavones inhibited inflammation, upregulated CES, and markedly enhanced the conversion of Lv into LvA. For the first time, we provide evidence that isoflavones and Lv in XZK act in concert to enhance the efficacy and reduce the side effects of Lv.
TL;DR: Candida glabrata is a useful model organism for examining resistance to statins and further studies are warranted to examine the precise molecular mechanisms of statin resistance.
Abstract: Reduced efficacy of statins has been observed in people but the mechanism of this resistance is unclear and no statin-resistance mutations in the catalytic domain of HMGCR have been reported. The present study focused on looking for statin-resistance mutations and examining the mechanism of statin resistance using Candida glabrata as a model organism. C. glabrata was cultured in media containing lovastatin, simvastatin or atorvastatin to obtain lovastatin-, simvastatin- and atorvastatin-resistant mutants. A single mutant from each was purified for further analysis. In each mutant, gene sequencing showed there were no changes in the catalytic domain of HMGCR. HMGCR was overexpressed in two resistant isolates suggesting that increased production of HMGCR can lead to resistance. In a third mutant, HMGCR activity was unaltered, suggesting a non-HMGCR related mechanism, such as increased drug efflux, could be operating. Candida glabrata is a useful model organism for examining resistance to statins. Further studies are warranted to examine the precise molecular mechanisms of statin resistance.
TL;DR: Lovastatin is the first FDA-approved antihypercholesterolemic drug for the treatment of cardiovascular diseases with pleiotropic clinical applications and Monascus purpureus is one of the safest molds for the production of lovastatin.
Abstract: Lovastatin is the first FDA-approved antihypercholesterolemic drug for the treatment of cardiovascular diseases with pleiotropic clinical applications. Monascus purpureus is one of the safest molds for the production of lovastatin that has been used in the preparation of Chinese medicine Red-Yeast Rice. This investigation dealt with the effect of both static and dynamic culture conditions on the morphological change and localization of lovastatin in Monascus purpureus. In dynamic culture condition, pellet morphology was observed and the maximum intra- and extra-cellular components of lovastatin including both β-hydroxy acid and lactone forms were found to be 1043.45 and 207.94 μg/l, respectively. Filamentous (mat) form of morphology was observed in the static culture condition and the intra- and extracellular concentration of lovastatin were 677.9 and 789.2 μg/l. Taguchi’s L12 (112) orthogonal arrays was employed to find optimal conditions for the submerged production of lovastatin and for the growth of Monascus purpureus. Three physical and five chemical variables were considered in the current experimental study. The maximum production of lovastatin was observed to be 3.66 mg/l. Among the chemical parameters, MnSO4 and MgSO4 were the most significant parameters for the production of lovastatin. Physical parameters, viz., agitation rate and temperature, were also equally significant to that of medium constituents for the production of lovastatin. The significance of the variables on the biomass growth was just opposite to that the results of lovastatin production. Ammonium chloride was the most significant parameter among the variables studied for the growth of M. purpureus, followed by glucose and phosphorous sources (KH2PO4). Statistical analysis showed that those parameters were significant with more than 99 % confidence (p<0.01).
TL;DR: Glucose and fructose assimilation kinetic parameters revealed that more lovastatin is produced during glucose assimilation, while more biomass was formed during fructose Assimilation.
Abstract: Lovastatin is a statin drug, which lowers cholesterol level in blood due to inhibition of (S)-3-hydroxy-3-methylglutaryl-CoA reductase. Date syrup is a rich medium for microbial growth and metabolite production. The main carbohydrates present in the date syrup are glucose and fructose. In this study, date syrup was used as a complex and bioresource medium for lovastatin production by Aspergillus terreus in the submerged cultivation. Optimization of the date syrup medium in order to achieve the highest titers of lovastatin and biomass was carried out. Four factors were studied by response surface methodology including concentration of date syrup carbohydrates, yeast extract concentration, pH, and rotation speed of the shaker. Optimal conditions for these factors found were as follows: concentration of date syrup carbohydrates, 64 g/l; yeast extract concentration, 15 g/l; pH, 6.5; and agitation speed, 150 rpm. It gave lovastatin concentration of 105.6 mg/l. Next, batch cultures in the optimal conditions were performed in a 2.5-l working volume bioreactor and led to the lovastatin titer of 241.1 mg/l during 12 days. Aspergillus terreus showed diauxic growth in the optimized medium with a shift from glucose to fructose assimilation during the run. Glucose and fructose assimilation kinetic parameters revealed that more lovastatin is produced during glucose assimilation, while more biomass was formed during fructose assimilation.