Simvastatin

Manufactured by Cayman Chemical

Sourced in United States

Simvastatin is a laboratory reagent manufactured by Cayman Chemical. It is a white crystalline solid that functions as an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, an enzyme involved in the production of cholesterol in the body.

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Lab products found in correlation

Atorvastatin, by Cayman Chemical (8 mentions) Fluvastatin, by Cayman Chemical (5 mentions) Pravastatin, by Cayman Chemical (4 mentions) Lovastatin, by Cayman Chemical (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Mevalonate, by Merck Group (3 mentions) Cholesterol, by Merck Group (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Cayman Chemical (2 mentions) Ay9944, by Cayman Chemical (2 mentions) Mg132, by Merck Group (2 mentions) Nsc23766, by Bio-Techne (2 mentions) Phosphatase inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Eht1864, by Cayman Chemical (2 mentions) Mouse mab 9139, by Cell Signaling Technology (2 mentions) L 788 123, by Cayman Chemical (2 mentions) Ehop 016, by Cayman Chemical (2 mentions) Water soluble cholesterol, by Merck Group (2 mentions) Geranylgeranyl pyrophosphate, by Merck Group (2 mentions)

34 protocols using simvastatin

Simvastatin treatment of A549 cells

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Human alveolar epithelial cells (A549) (ATCC, Manassas, VA) were maintained in Dulbecco's Modified Eagle's Medium (DMEM) (Corning, Manassas, VA), supplemented with 10% Fetal Bovine Serum (FBS) (Thermo Scientific, Rockford, IL) and 1% of antibiotics/antimycotics mixture (Life Technologies, Grand Island, NY) at 37°C, 5% CO₂ and 95% relative humidity. Cells were seeded onto 40 mm diameter glass cover slips inside 60 mm petri dishes at a cell density of 3.6 x 10⁴ cells/cm² and grown to confluence. Based on previous studies which treated lung epithelial or endothelial cells with 0.1 to 100μM Simvastatin [29 (link), 31 (link)–33 (link)], in this study, A549 cells were incubated with 2.5 to 50μM Simvastatin (Cayman Chemical, Ann Arbor, MI) in supplemented media for 16–17 hours prior to each experiment and untreated cells were used as controls.

Higuita-Castro N., Shukla V.C., Mihai C, & Ghadiali S.N. (2016). Simvastatin Treatment Modulates Mechanically-Induced Injury and Inflammation in Respiratory Epithelial Cells. Annals of biomedical engineering, 44(12), 3632-3644.

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Optimizing Zebrafish Larvae Diet Exposure

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To optimize a protocol for exposing larvae to HF and HFC diets, we first assessed the consumption of control diet by zebrafish larvae. We fed 100 larvae in a 2 L fish tank with 30 mg-180 mg per day to meet their basic energy requirement. We found that the incidence of steatosis in zebrafish larvae was gradually increased with the growing amount of feeding. Based on these results, all types of diets given to larvae were strictly restricted as 30 mg/tank per day. Ezetimibe (Santa Cruz Biotechnology, Texas, USA) used in this study was added directly to the fish tank water at concentrations of 1 μM and simvastatin (Cayman Chemicals, Michigan, USA) was added at 50 μg/g food. These concentrations of drugs and methods of administration were chosen based on previous report as described [34 (link)].

Dai W., Wang K., Zheng X., Chen X., Zhang W., Zhang Y., Hou J, & Liu L. (2015). High fat plus high cholesterol diet lead to hepatic steatosis in zebrafish larvae: a novel model for screening anti-hepatic steatosis drugs. Nutrition & Metabolism, 12, 42.

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Cholesterol Synthesis Inhibition in Cell Lines

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HEK293T (parental and hCLTA^EN-Tq2) and human SK-MEL-2 (hCLTA^EN-Tq2 hDNM2^EN-eGFP, as previously described; Scott et al., 2018 (link)) were maintained in DMEM (GIBCO, 4.5 g/L glucose, 110 mg/L pyruvate), supplemented with 10% (v/v) FBS (Hyclone), and 1,000 U/mL penicillin/streptomycin. For inhibition of cholesterol synthesis, cells were rinsed twice in serum-free DMEM and cultured under cholesterol deplete conditions in 7.5% delipidated serum (LPDS) for 48 h with small molecule inhibitors AY9944 (2.5 μM, Cayman Chemical; DHCR7 inhibitor), U18666A (20 nM, Cayman Chemical; DHCR24 inhibitor), Atorvastatin (1 μM, Cayman Chemical), or Simvastatin (Cayman Chemical), or at concentrations as otherwise specified. Acute sterol depletion was achieved by 1 h incubation with 5 mM MβCD (Alfa Aesar, 1303.31 g/mol) at 37°C. De-identified fibroblasts cultured from skin punch biopsies from Smith-Lemli-Opitz subjects (kind gift from Dr. Forbes Porter, NICHD) were maintained in DMEM (GIBCO, 4.5 g/L glucose, 110 mg/L pyruvate), supplemented with 15% (v/v) FBS (Hyclone), and 1,000 U/mL penicillin/streptomycin. SLOS fibroblasts were rinsed twice in serum-free DMEM and cultured for 10 days in 7.5% (v/v) LPDS to induce biochemical profiles. SLOS fibroblast cells lines were described previously (Krakowiak et al., 2000 (link); Wassif et al., 1998 (link)).

Anderson R.H., Sochacki K.A., Vuppula H., Scott B.L., Bailey E.M., Schultz M.M., Kerkvliet J.G., Taraska J.W., Hoppe A.D, & Francis K.R. (2021). Sterols lower energetic barriers of membrane bending and fission necessary for efficient clathrin-mediated endocytosis. Cell reports, 37(7), 110008.

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Simvastatin and Estrogen Preparation

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Simvastatin (Cayman Chemical, Ann Arbor, MI) stock solution (10 mM) was prepared in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO) and stored at −20 °C until use. Estrogen was purchased from Sigma-Aldrich, and the stock solution (10 mM) was prepared in ethanol (Sigma-Aldrich). The rest of the chemicals and reagents used in the experiments were purchased from Thermo Fisher Scientific and Sigma-Aldrich.

Afrin S., El Sabeh M., Islam M.S., Miyashita-Ishiwata M., Malik M., Catherino W.H., Akimzhanov A.M., Boehning D., Yang Q., Al-Hendy A., Segars J.H, & Borahay M.A. (2021). Simvastatin modulates estrogen signaling in uterine leiomyoma via regulating receptor palmitoylation, trafficking and degradation. Pharmacological research, 172, 105856.

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Simvastatin Nanoparticle Formulation Protocol

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Simvastatin was purchased from Cayman Chemical (Ann Arbor, MI). Simvastatin was activated prior to its use as previously described (Borahay et al., 2015 ). Activation converts the prodrug (lactone structure) to its active form (β-hydroxyacid structure). Briefly, Simvastatin (25 mg) was dissolved in 625μL of ethanol. This solution was then added to 935μL of 0.1 NaOH and heated in a water bath (50°C) for 2 hours before diluting with water. The pH was adjusted to reach 7 using HCl. This solution was diluted 8-fold with water to reach a 2mg/mL concentration and was sterile filtered before being stored at 4°C until use.
Simvastatin-loaded liposome nanoparticles (Simvastatin-NP) was prepared using 1,2-Dioleoyl-sn-Glycero-3-Phosphocholine (DOPC) which were purchased from Avanti Polar Lipids (Alabaster, AL) and 1,2-Distearoyl-sn-glycero-3-phosphorylethanolamine (DSPE). DOPC/DSPE-Peg2000 were prepared in 10:1 mixture and Simvastatin-DOPC/DSPE-Peg2000 was prepared in 1:10 mixture. Simvastatin-DOPC/DSPE-Peg2000 was kept lyophilized at −20°C and dissolved in normal saline immediately prior to use.

El Sabeh M., Vincent K.L., Afrin S., Motamedi M., Saada J., Yang J., Ozpolat B., Kilic G.S, & Borahay M.A. (2022). Simvastatin-loaded Liposome Nanoparticles Treatment for Uterine Leiomyoma in a Patient-Derived Xenograft Mouse Model: A Pilot Study. Journal of obstetrics and gynaecology : the journal of the Institute of Obstetrics and Gynaecology, 42(6), 2139-2143.

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PLGA Nanoparticle Formulation and Characterization

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Ester-terminated poly(lactic-co-glycolic)acid (PLGA 85:15) was obtained from Lactel (Birmingham, AL, USA). Dichloromethane (DCM), acetonitrile (MeCN), dimethyl sulfoxide (DMSO), poly(vinyl alcohol) (PVA, 31,000–50,000 Da, 87–89% hydrolyzed), chitosan (low molecular weight (50,000–190,000 Da), 75–85% deacetylated) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Simvastatin was obtained from Cayman Chemical (Ann Arbor, MI, USA). Fetal bovine serum (FBS), penicillin-streptomycin, amphotericin B were from Life Technologies (Grand Island, NY, USA), 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate salt (DiD) was from Thermofisher Scientific (Waltham, MA, USA) and cOmpleteTM protease inhibitor cocktail was obtained from Roche (Basel, Switzerland).

Yang F., Cabe M., Nowak H.A, & Langert K.A. (2022). Chitosan/poly(lactic-co-glycolic)acid Nanoparticle Formulations with Finely-Tuned Size Distributions for Enhanced Mucoadhesion. Pharmaceutics, 14(1), 95.

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Preparation of Statin Compounds

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Simvastatin, Pravastatin, Lovastatin, Atorvastatin and Fluvastatin (Cayman Chemical, Ann Arbor, MI, USA) stock solutions (10 mM) were prepared in dimethyl sulfoxide (DMSO; Sigma‐Aldrich) and stored at −20°C until use. The rest of the chemicals and reagents used in the experiments were purchased from Sigma‐Aldrich and Thermo Fisher Scientific.

Afrin S., Ali M., El Sabeh M., Yang Q., Al‐Hendy A, & Borahay M.A. (2022). Simvastatin inhibits stem cell proliferation in human leiomyoma via TGF‐β3 and Wnt/β‐Catenin pathways. Journal of Cellular and Molecular Medicine, 26(5), 1684-1698.

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Evaluating Cell Cycle Regulators

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Simvastatin was purchased from Cayman (Ann Arbor, MI, USA) and propidium iodide (PI), CHX, MG132 and mevalonate were obtained from Sigma (St. Louis, MO, USA). TRIzol reagent was obtained from Invitrogen (Carlsbad, CA, USA). Antibodies specific to p21, p27, cyclin D1, cyclin E1, phospho-AMPK, AMPK, phospho-STAT3 and STAT3 were purchased from Cell Signaling Technology (Danvers, MA, USA), the antibody specific to p45^SKP2 was purchased from Invitrogen, and the antibody specific to β-actin was purchased from Santa Cruz (Santa Cruz, CA, USA).

Wang S.T., Ho H.J., Lin J.T., Shieh J.J, & Wu C.Y. (2017). Simvastatin-induced cell cycle arrest through inhibition of STAT3/SKP2 axis and activation of AMPK to promote p27 and p21 accumulation in hepatocellular carcinoma cells. Cell Death & Disease, 8(2), e2626-.

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Enhancing NK-92 Cell Cytotoxicity with Statins and Supplements

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NK-92 cells were seeded at 0.1 × 10⁶ cells/mL in round-bottom 96-well plates (3799, Corning Life Sciences B.V., Amsterdam, the Netherlands). Atorvastatin (10493, Cayman Chemical), fluvastatin (10010337, Cayman Chemical), pravastatin (10010342, Cayman Chemical), rosuvastatin (12029, Cayman Chemical), and simvastatin (10010344, Cayman Chemical) were dissolved in DMSO (Sigma-Aldrich). Statins were used at final concentration of 20 μM, 5 μM, and 0.5 μM. DMSO was diluted in the same volume and served as solvent control. IL-2, IL-21 (Thermo Fisher Scientific), vitamin C (Sigma-Aldrich), dextran (Sigma-Aldrich), prostaglandin E2 (PGE2, Sigma-Aldrich), protamine sulfate (Sigma-Aldrich), and vectofusin-1 (Miltenyi Biotec) were dissolved in distilled water (GIBCO). DMSO was added 0.8 μL, 0.2 μL, and 0.02 μL, respectively, at the same volume as the statin group in 96-well plates. IL-21 was added at 20 ng/mL, 5 ng/mL, and 0.5 ng/mL. vitamin C was used at concentrations of 500 μg/mL, 50 μg/mL, and 5 μg/mL as described previously.²³ (link) dextran was used at 80 μg/mL, 8 μg/mL, and 0.8 μg/mL.¹⁵ PGE2 was used at 100 μM, 10 μM, and 1 μM.²² (link) vectofusin-1 was used at 50 μg/mL, 5 μg/mL, and 0.5 μg/mL.¹⁴ (link)^,⁶¹ (link) GGPP ammonium salt was purchased from Sigma-Aldrich and was added at 10 μM in co-culture assays.²⁹ (link)

Gong Y., Klein Wolterink R.G., Janssen I., Groot A.J., Bos G.M, & Germeraad W.T. (2020). Rosuvastatin Enhances VSV-G Lentiviral Transduction of NK Cells via Upregulation of the Low-Density Lipoprotein Receptor. Molecular Therapy. Methods & Clinical Development, 17, 634-646.

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Statin Drugs Mevalonate Pathway Inhibitors

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Statin drugs were obtained from commercial sources as follows: pitavastatin calcium from Atomole, lovastatin, pravastatin sodium and atorvastatin calcium from Toronto Research Chemicals, simvastatin from Cayman Chemical and rosuvastatin calcium from Biotang. Mevalonate pathway metabolites R-mevalonic acid sodium salt and farnesyl pyrophosphate ammonium salt were from Sigma and geranyl pyrophosphate ammonium salt was from Axon MedChem. PARP1/2 inhibitor veliparib (ABT-888) was obtained from Chemie Tek.

Efimova E.V., Ricco N., Labay E., Mauceri H.J., Flor A.C., Ramamurthy A., Sutton H.G., Weichselbaum R.R, & Kron S.J. (2017). HMG-CoA reductase inhibition delays DNA repair and promotes senescence after tumor irradiation. Molecular cancer therapeutics, 17(2), 407-418.

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