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Dl thiorphan

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DL-thiorphan is a laboratory reagent used in scientific research. It is a neutral aminopeptidase inhibitor. The core function of DL-thiorphan is to inhibit the breakdown of certain peptides in experimental settings.

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Lab products found in correlation

Bestatin hcl, by Merck Group (4 mentions) Naloxone, by Bio-Techne (2 mentions) Naloxone, by Merck Group (2 mentions) Met enkephalin acetate salt, by Merck Group (2 mentions) Mk 801, by Merck Group (2 mentions) Lisinopril, by Merck Group (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Kanamycin, by Merck Group (1 mentions) L captopril, by Merck Group (1 mentions) L arginine, by Merck Group (1 mentions) Picrotoxin, by Merck Group (1 mentions) Tetrodotoxin, by Bio-Techne (1 mentions) Strychnine, by Merck Group (1 mentions) Ml297, by Bio-Techne (1 mentions) 4 aminopyridine 4 ap, by Merck Group (1 mentions) Bestatin, by Merck Group (1 mentions) Dynorphin a, by Merck Group (1 mentions) Nor bni, by Bio-Techne (1 mentions) Heparin coated microvettes, by Sarstedt (1 mentions) M1000 pro, by Tecan (1 mentions)

Spelling variants of this product

Thiorphan (17 citations)

15 protocols using dl thiorphan

1

Pharmacological Modulation of Cocaine Behaviors

Cited 1 time
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Doxycycline was administered ad libitum as a supplemented chow (40 mg/kg, Mutual Pharmaceutical) for 2 weeks to switch the transgene off. Cocaine (15 mg/kg, Sigma) and saline were administered intraperitoneally (i.p.) at 10 m//kg body weight. Naloxone, met-enkephalin acetate salt, DL-thiorphan, and bestatin HCl were supplied by Millipore Sigma. Gabazine (SR95531) was supplied by Abcam and CTAP and Naloxone were from Tocris. Doses for cocaine and Naloxone for place conditioning were selected based on previous experiments (Cunningham et al., 1995 (link); Dobbs et al., 2016 (link); Skoubis et al., 2005 (link); Solecki et al., 2009 (link)). The in vivo met-enkephalin dose was selected based on our ex vivo slice physiology experiments. The CTAP dose was selected based off efficacy in blocking cocaine preference and opioid analgesia in mice and rats (Kramer et al., 1989 ; Soderman and Unterwald, 2008 (link)). Drugs were dissolved in aCSF (electrophysiology) or saline (CPP).
Dai K.Z., Choi I.B., Levitt R., Blegen M.B., Kaplan A.R., Matsui A., Shin J.H., Bocarsly M.E., Simpson E.H., Kellendonk C., Alvarez V.A, & Dobbs L.K. (2022). Dopamine D2 receptors bidirectionally regulate striatal enkephalin expression: Implications for cocaine reward. Cell reports, 40(13), 111440.
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2

Pharmacological Modulation of Cocaine Behaviors

Cited 1 time
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Doxycycline was administered ad libitum as a supplemented chow (40 mg/kg, Mutual Pharmaceutical) for 2 weeks to switch the transgene off. Cocaine (15 mg/kg, Sigma) and saline were administered intraperitoneally (i.p.) at 10 m//kg body weight. Naloxone, met-enkephalin acetate salt, DL-thiorphan, and bestatin HCl were supplied by Millipore Sigma. Gabazine (SR95531) was supplied by Abcam and CTAP and Naloxone were from Tocris. Doses for cocaine and Naloxone for place conditioning were selected based on previous experiments (Cunningham et al., 1995 (link); Dobbs et al., 2016 (link); Skoubis et al., 2005 (link); Solecki et al., 2009 (link)). The in vivo met-enkephalin dose was selected based on our ex vivo slice physiology experiments. The CTAP dose was selected based off efficacy in blocking cocaine preference and opioid analgesia in mice and rats (Kramer et al., 1989 ; Soderman and Unterwald, 2008 (link)). Drugs were dissolved in aCSF (electrophysiology) or saline (CPP).
Dai K.Z., Choi I.B., Levitt R., Blegen M.B., Kaplan A.R., Matsui A., Shin J.H., Bocarsly M.E., Simpson E.H., Kellendonk C., Alvarez V.A, & Dobbs L.K. (2022). Dopamine D2 receptors bidirectionally regulate striatal enkephalin expression: Implications for cocaine reward. Cell reports, 40(13), 111440.
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3

Neprilysin Activity in Mouse Plasma

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Lateral saphenous vein blood from conscious, non-fasted mice was collected following eight weeks of treatment. Plasma neprilysin activity was assessed by a fluorometric enzyme method, as previously described (11 (link)). Each plasma sample was assayed in both the absence and presence of a specific neprilysin inhibitor (DL-thiorphan, Sigma-Aldrich, St Louis, MO, USA) to differentiate neprilysin activity from non-specific endopeptidase activity.
Esser N., Schmidt C., Barrow B.M., Cronic L., Hackney D.J., Mongovin S.M., Hogan M.F., Templin A.T., Castillo J.J., Hull R.L, & Zraika S. (2022). Insulinotropic Effects of Neprilysin and/or Angiotensin Receptor Inhibition in Mice. Frontiers in Endocrinology, 13, 888867.
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4

Chemical Reagents for Biochemical Assays

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l-Arginine, dl-thiorphan, l-captopril, and kanamycin were obtained from Sigma-Aldrich
(St. Louis, MO). Thanatin (GSKKPVPIIYCNRRTGKCQRM) was purchased from
GL Biochem (Shanghai, CN). d-Captopril was purchased from
TLC Pharmaceutical Standards (Pony Dr, CA). Fluorsecein-5-maleimide
was obtained from Invitrogen (Carlsbad, CA, USA).
Chung S.F., Tam S.Y., Wong W.T., So P.K., Cheong W.L., Mak C.W., Lee L.M., Chan P.H., Wong K.Y, & Leung Y.C. (2024). Fluorescently Modified NDM-1: A Versatile Drug Sensor for Rapid In Vitro β-Lactam Antibiotic and Inhibitor Screening. ACS Omega, 9(8), 9161-9169.
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5

Pharmacological Modulation of Respiratory Circuits

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ME ([Met5]-enkephalin acetate salt), bestatin, DL-thiorphan, strychnine, picrotoxin, DNQX, 4-aminopyridine (4AP), and MK801 were from Sigma-Aldrich (St Louis, MO). Tetrodotoxin and ML-297 was from Tocris Bio-Techne (Minneapolis, MN). All drugs were applied by bath perfusion. bestatin (10 µM) and thiorphan (1 µM) were included with ME to prevent degradation. ME is an endogenous opioid peptide agonist for mu and delta opioid receptors. Delta opioid receptors are not expressed in KF or preBötC neurons (Varga et al., 2020 (link)) and do not cause opioid-induced respiratory depression (Dahan et al., 2001 (link)). An EC80 concentration of ME (1–3 µM) was used to ensure robust and reliable responses but avoid acute receptor desensitization that occurs with higher concentrations (Levitt and Williams, 2018 (link)).
Bateman J.T, & Levitt E.S. (2023). Opioid suppression of an excitatory pontomedullary respiratory circuit by convergent mechanisms. eLife, 12, e81119.
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6

Dynorphin A-(1-17) Receptor Activation

Cited 2 times
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The endogenous kappa-opioid receptor agonist, dynorphin A-(1–17) (Tocris) was bath applied for 10 min at a concentration of 300 nM as previously reported [29 (link)]. Endopeptidase inhibitors bestatin HCl (10 μM, Sigma-Aldrich) and DL-Thiorphan (1 μM, Sigma-Aldrich) were combined with dynorphin A [37 (link)]. A previous study has shown that KOR agonism is not reversed by the KOR antagonist Nor-Binaltorphimine dihydrochloride (nor-BNI) [29 (link)]. Thus, nor-BNI (100 nM, Tocris) was added to the extracellular aCSF solution and pre-applied to slices for at least 15 min prior to experiments. For chemogenetic experiments, salvinorin B (SalB; Tocris) was bath applied for 10 min at a 100 nM concentration [38 (link)].
Pina M.M., Pati D., Hwa L.S., Wu S.Y., Mahoney A.A., Omenyi C.G., Navarro M, & Kash T.L. (2019). The kappa opioid receptor modulates GABA neuron excitability and synaptic transmission in midbrain projections from the insular cortex. Neuropharmacology, 165, 107831.
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7

Neprilysin Activity Measurement in Plasma

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Plasma neprilysin activity was measured as described (11 (link)). Briefly, blood plasma was separated by centrifugation from whole blood collected on heparin-coated microvettes (Sarstedt, Nümbrecht, Germany). Plasma samples in triplicate were incubated at 37°C with the substrate glutaryl-Ala-Ala-Phe-4-methoxy-2-naphthylamine (Sigma-Aldrich, St. Louis, Missouri) and aminopeptidase M (R and D Systems, Minneapolis, Minnesota) to generate the fluorescent product methoxy-2-naphthlamine (MNA) in the absence or presence of a specific neprilysin inhibitor (DL-Thiorphan, Sigma-Aldrich). Fluorescence was read on a Tecan M1000 Pro multimode spectrophotometer (ex/em 340/420 nm, Tecan, Männedorf, Switzerland), compared against a MNA standard curve, and expressed as pmol MNA/h/uL plasma.
Croteau D., Qin F., Chambers J.M., Kallick E., Luptak I., Panagia M., Pimentel D.R., Siwik D.A, & Colucci W.S. (2020). Differential Effects of Sacubitril/Valsartan on Diastolic Function in Mice With Obesity-Related Metabolic Heart Disease. JACC: Basic to Translational Science, 5(9), 916-927.
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8

Morphine Sulfate Receptor Assay

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Morphine sulfate was obtained from the National Institute on Drug Abuse Drug Supply Program (RTI International, Research Triangle Park, NC). ME ([Met5]-enkephalin acetate salt), bestatin HCl, DL-thiorphan and MK801 were from Sigma-Aldrich (St. Louis, MO). (RS)-baclofen was from Tocris (Minneapolis, MN).
Varga A.G., Reid B.T., Kieffer B.L, & Levitt E.S. (2019). Differential impact of two critical respiratory centers in opioid-induced respiratory depression in awake mice. The Journal of physiology, 598(1), 189-205.
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9

In vitro study of osteosarcoma cell lines

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Human osteosarcoma cell lines HOS (ATCC, no. CRL-1543TM, ATCC, Manassas, VA, USA) and Saos-2 (ATCC, no. HTB-85), both derived from primary bone tumors and human normal cells—colon fibroblasts CCD-18Co (ATCC, no. CRL-1459TM) and fetus osteoblasts hFOB 1.19 (ATCC, no. CRL-11372TM) were used in the present study. The cells were cultured in appropriate media: HOS and CCD-18Co—EMEM (Sigma Aldrich, St. Louis, MO, USA), Saos-2—McCoy’s (Sigma Aldrich), and hFOB 1.19—DMEM/F-12 without phenol red and with L-glutamine. All media were supplemented with fetal bovine serum (FBS, Sigma Aldrich). The HOS and Saos-2 culture media were supplemented with an antibiotic/antimycotic solution (Sigma Aldrich), whereas the hFOB1.19 medium was additionally supplemented with G418 (Sigma Aldrich); the CCD-18Co medium was used without antibiotics. The cells were cultivated in standard conditions at 37 °C, 95% humidity, and with 5% CO2, except for the hFOB 1.19 cells, which required 34 °C.
All of the tested compounds (1–7) were synthesized with the methods described previously [23 (link),24 ]. The stock solutions of these compounds were prepared in DMSO, whereas their working solutions at the required concentrations were made by dilution in the appropriate culture medium.
The solutions of DL-Thiorphan (Sigma-Aldrich) were prepared as described previously [9 (link)].
Mizerska-Kowalska M., Sowa S., Donarska B., Płaziński W., Sławińska-Brych A., Tomasik A., Ziarkowska A., Łączkowski K.Z, & Zdzisińska B. (2022). New Borane-Protected Derivatives of α-Aminophosphonous Acid as Anti-Osteosarcoma Agents: ADME Analysis and Molecular Modeling, In Vitro Studies on Anti-Cancer Activities, and NEP Inhibition as a Possible Mechanism of Anti-Proliferative Activity. International Journal of Molecular Sciences, 23(12), 6716.
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10

Neprilysin and DPP-4 Activities in Mice

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Neprilysin activity was determined in plasma samples taken from mice following 2, 6, 10, 12, 14 and 16 weeks of feeding. Lateral saphenous vein blood from conscious, fed mice was collected in heparinised tubes and centrifuged to obtain plasma, which was stored at −30°C prior to assay. In the neprilysin activity assay, glutaryl-Ala-Ala-Phe-4-methoxy-2-naphthylamine is broken down by neprilysin in plasma to phe-4-methoxy-2-naphthylamine, and then by aminopeptidase M to the fluorescent product methoxy-2-naphthylamine [14 (link)]. Each plasma sample was assayed in both the absence and presence of a specific neprilysin inhibitor (dl-thiorphan, Sigma-Aldrich, St Louis, MO, USA) to distinguish neprilysin activity from non-specific endopeptidase activity. Fluorescence was compared against a methoxy-2-naphthylamine standard curve.
DPP-4 activity was determined in plasma samples obtained from mice immediately after the OGTT. Arterial (carotid) blood was collected in heparinised tubes and centrifuged to obtain plasma, which was stored at −30°C prior to assay. In this assay, DPP-4 cleaves non-fluorescent H-Gly-Pro-7-amino-4-methylcoumarin to generate fluorescent 7-amino-4-methylcoumarin [15 (link)]. Plasma DPP-4 activity was compared against a 7-amino-4-methylcoumarin standard curve.
Willard J.R., Barrow B.M, & Zraika S. (2016). Improved glycaemia in high-fat fed neprilysin-deficient mice is associated with reduced DPP-4 activity and increased active GLP-1 levels. Diabetologia, 60(4), 701-708.
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