L dopa

Manufactured by Cayman Chemical

Sourced in United States

L-DOPA is a chemical compound used as a laboratory reagent. It serves as a precursor for the synthesis of various biologically active compounds. The core function of L-DOPA is to provide a building block for further chemical reactions and analyses.

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7 protocols using l dopa

Tyrosinase Inhibition Assay Protocol

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Extracts were added to a 100-µL final volume containing five units of tyrosinase from mushrooms (Sigma T3824-25KU) in 50 mM monobasic potassium phosphate buffer, pH 6.5 at 25 °C. After 5 min of incubation at room temperature, an initial absorbance at 477 nm was recorded and 110 µL of 12 mM L-DOPA (#CAYM13248-5, Cayman Chemical) in the same buffer were added. Then, an absorbance at 477 nm was recorded every 15 s for 15 min. Bioinactive and bioactive controls were based on wells with or without tyrosinase, respectively. For data analysis, absorbance values at reading time giving the highest Z’ factor [25 (link)] were considered, and the initial absorbance value was subtracted from the time-related absorbance value for each well. Results are expressed as percentages of tyrosinase inhibition relative to the bioinactive controls (with tyrosinase, i.e., 0% inhibition) and bioactive controls (no tyrosinase, i.e., 100% inhibition).

Gori A., Boucherle B., Rey A., Rome M., Barette C., Soleilhac E., Philouze C., Fauvarque M.O., Fuzzati N, & Peuchmaur M. (2023). Investigation of Chemical Composition and Biological Activities of Ajuga pyramidalis—Isolation of Iridoids and Phenylethanoid Glycosides. Metabolites, 13(1), 128.

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Neurophysiological Modulation Protocol

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IVM (cat. no. NDC 55529-012-01, Norbrook Laboratories, Ltd, Newry, North Ireland, UK), Nicotine (cas. no. 54-11-5, Sigma-Aldrich, St. Louis, Missouri, USA), Hexamethonium bromide (cas. no. 55–97-0, Cayman Chemical Company, Ann Arbor, Michigan, USA), L-DOPA (cat. no. PHR1271, Sigma-Aldrich, St. Louis, Missouri, USA), and 5-(3-Bromophenyl)-1,3-dihydro-2H-benzofuro[3,2-e]-1,4-diazepin-2-one (5-BDBD) (cat. no. T22518, TargetMol, Wellesley Hills, Massachusetts, USA) were dissolved in stock solutions and then diluted into ACSF at specified concentrations (0.1–100 µM IVM, 300 nM Nicotine, 200 µM Hexamethonium, 10 µM L-DOPA, and 10 µM 5-BDBD). Drugs brain slice administration used either gravity-based flow system or peristaltic pumps (1–2 ml/min).

Wadsworth H.A., Warnecke A.M., Barlow J.C., Robinson J.K., Steimle E., Ronström J.W., Williams P.E., Galbraith C.J., Baldridge J., Jakowec M.W., Davies D.L, & Yorgason J.T. (2024). Ivermectin increases striatal cholinergic activity to facilitate dopamine terminal function. Cell & Bioscience, 14, 50.

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Molecular Mechanisms of L-dopa Action

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L-dopa was purchased from Cayman Chemical (Ann Arbor, MI, USA). The following antibodies were used: anti-SYT12 (M519) (Merck & Co., Inc., Kenilworth, NJ, USA); anti-CAMK2N1 (PA5-23740) (Invitrogen, CA, USA); anti-CAMK2 (ab52476) and anti-CAMK2 (Phospho T286) (ab32678) (abcam, Tokyo, Japan); anti-p27^Kip1 (#3686) and anti-CDK2 (#2546) (Cell Signaling Technology, Danvers, MA, USA); anti-cyclin E (sc-377100), and GAPDH (sc-3223) (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Eizuka K., Nakashima D., Oka N., Wagai S., Takahara T., Saito T., Koike K., Kasamatsu A., Shiiba M., Tanzawa H, & Uzawa K. (2019). SYT12 plays a critical role in oral cancer and may be a novel therapeutic target. Journal of Cancer, 10(20), 4913-4920.

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Diverse Compound Screening Protocol

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Purchased compounds included thiostrepton (Cayman#19200), losartan potassium (Sigma#61188), sulfathiazole (Sigma#S9876), N-laurylsarcosine (Sigma#L7414), captopril (Sigma#C4042) atracurium besylate (Cayman#17796), L-DOPA (Cayman#13248), idoxuridine (Cayman#20222), dyphylline (Cayman#17958), dapsone (Cayman#23743), pyrimethamine (Cayman#16472), hydroflumethiazide (Cayman#23612), recombinant human TGFβ1 (Peprotech#100-21), SB-431542 (Sigma#S4317). All compounds were suspended in dimethyl sulfoxide (DMSO), and final vehicle concentration was <0.1% (v/v).

Salvi A., Hardy L.R., Heath K.N., Watry S., Pergande M.R., Cologna S.M, & Burdette J.E. (2022). PAX8 modulates the tumor microenvironment of high grade serous ovarian cancer through changes in the secretome. Neoplasia (New York, N.Y.), 36, 100866.

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Ceftriaxone Neuroprotection in Parkinson's

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The overall study design is illustrated in Figure 1. On day 7 post-6-OHDA lesion, Ceftriaxone (200 mg/kg, i.p.) or saline was administered for 7 consecutive days and every other week (days 7–13, days 21–27 and 35–39). Ceftriaxone (disodium salt hemiheptahydrate) (Hospira, Lake Forest, IL or Toronto Research Chemicals, Inc., Toronto, ON, cat#C245000) was dissolved in sterile saline (0.9%) and prepared daily. The dose of L-DOPA (Sigma, cat # D9628) at one experiment site was 12 mg/kg L-DOPA and 6 mg/kg L-DOPA (methyl ester) (Cayman Chemical, act#16149) was used at the two subsequent studies. In combination with 15 mg/kg benserazide-HCl, i.p., L-DOPA was given for 20 consecutive days, starting on day 19 (day 1 of L-DOPA) until day 39 post-lesion. No significant difference in total ALO was observed between using either dose of L-DOPA in either the L-DOPA alone (experiment site, F_(1,5) =1.55, p=0.27; days of L-DOPA, F_(5,25) =26.0, p<0.0001) or L-DOPA + Ceftriaxone group (experiment site, F_(1,11) =1.09, p=0.32; days of L-DOPA, F_(5,55) =2.44, p<0.05) and therefore study outcomes were collapsed between study sites.

Chotibut T., Meadows S., Kasanga E., McInnis T., Cantu M.A., Bishop C, & Salvatore M.F. (2017). Ceftriaxone reduces L-DOPA-induced dyskinesia severity in 6-OHDA Parkinson’s disease model. Movement disorders : official journal of the Movement Disorder Society, 32(11), 1547-1556.

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Pharmacological Characterization of PDM-042

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L-DOPA (Cayman Chemical Company, Ann Arbor, MI, USA) and benserazide (Sigma-Aldrich, Saint Louis, MO, USA) were dissolved in sterile 0.9% saline solution and administered subcutaneously (s.c., 2 mL/kg). The PDE10A inhibitor PDM-042 (synthesized at Mochida Pharmaceutical Co., Tokyo, Japan) was dissolved in 0.9% saline containing 0.5% methylcellulose and administered by gavage (5 mL/kg). All other reagents were of the highest grade commercially available. The pharmacological characterization of PDM-042 ((E)-4-(2-(2-(5,8-dimethyl-[1 (link),2 (link),4 (link)]triazolo [1,5-a]pyrazin-2-yl)vinyl)-6-(pyrrolidin-1-yl)pyrimidin-4-yl)morpholine) was described before [20 (link)]. PDM-042 demonstrated high brain penetration (striatum/plasma ratio = 6.3). The occupancy rate of PDE10A for PDM-042 at its ED50 (0.44 mg/kg) was 39%. The occupancy rates of PDE10A at the doses used of 1 and 3 mg/kg were 66% and 86.6%. Furthermore, PDM-042 showed good oral bioavailability (33%) [20 (link)]. Thus, PDM-042 is a highly powerful, selective, orally active, and brain penetrant PDE10A inhibitor.

Guimarães R.P., Ribeiro D.L., Dos Santos K.B., Talarico C.H., Godoy L.D, & Padovan-Neto F.E. (2022). Phosphodiesterase 10A Inhibition Modulates the Corticostriatal Activity and L-DOPA-Induced Dyskinesia. Pharmaceuticals, 15(8), 947.

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Investigating Neuroprotective Compounds

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N-acetylcysteine amide (NACA) was obtained from TOCRIS. Taurordeoxycholic acid (TUDCA) and L-dopa were purchased from Cayman Chemicals and taurine was purchased from Sigma (St. Louis, MO, USA). A stock solution of TUDCA was prepared in dimethylsulfoxide (DMSO). All other drug stocks were prepared in sterile water.

Sarkar H., Lahne M., Nair N, & Moosajee M. (2023). Oxidative and Endoplasmic Reticulum Stress Represent Novel Therapeutic Targets for Choroideremia. Antioxidants, 12(9), 1694.

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