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9 protocols using benzydamine hydrochloride

1

Synthesis and Characterization of Pharmaceutical Compounds

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Acetonitrile, ampicillin, flavin adenine dinucleotide disodium salt hydrate (FAD), benzydamine hydrochloride, tamoxifen, clomiphene citrate salt, NADPH, Isopropyl-ß-D-thiogalactopyranoside (IPTG), IGEPAL (octylphenoxy poly(ethyleneoxy)ethanol) and potassium phosphate were purchased from Sigma-Aldrich.
The drugs under study were purchased from different sources, clomiphene from DBA Italia (Italy); tozasertib from Aurogene (Rome, Italy); dasatinib from Selleck Chemicals (Houston, USA) and GSK5182 (4-[1-[4-[2-(Dimethylamino)ethoxy]phenyl]-5-hydroxy-2-phenylpent-1-enyl]phenol) (with isomer ratio of 1:1, E:Z) from Glaxosmithkline (UK).
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2

Biopolymer-Based Delivery System for Pharmaceuticals

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Poly (DL-lactic acid) with inherent viscosity 0.55–0.75 dL/g was supplied from Lactel Absorbable Polymers (Birmingham, AL, USA). Chitosan (low molar mass, degree of deacetylation > 75%), casein sodium salt from bovine milk, benzydamine hydrochloride, betahistine dihydrochloride, mucin (type II, from porcine stomach) and Bradford reagent were purchased from Sigma-Aldrich GmbH (Taufkirchen, Germany). Tolfenamic acid was delivered by Cayman Chemical Europe (Tallinn, Estonia). All other chemicals were of analytical grade.
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3

High-Throughput Cytochrome P450 Profiling

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Acetaminophen, astemizole, bupropion, N‐acetyl‐p‐aminobenzoic acid, 4‐aminobenzoic acid, benzydamine hydrochloride, chlorzoxazone, coumarin, dextromethorphan hydrobromide, 7‐Ethoxyresorufin, 7‐Hydroxycoumarin β‐D‐glucuronide sodium salt, 7‐Hydroxycoumarin sulfate potassium salt, 6β‐hydroxy testosterone, irinotecan, midazolam, phenacetin, resorufin, SN38, and sulfamethazine were purchased from Sigma Aldrich. Dextrorphan tartrate, diclofenac sodium salt, 4‐hydroxydiclofenac, hydroxy bupropion, S‐mephenytoin, 4‐hydroxyquinoline, paclitaxel, and testosterone were purchased from Cayman Chemical. 7‐hydroxycoumarin was purchased from Chem Service. Benzydamine N‐oxide, kynuramine hydrobromide, and N‐acetyl sulfamethazine were obtained from Santa Cruz Biotechnology. Carbazeran, 4‐hydroxyCarbazeran, 6‐hydroxychlorzoxazone, 6α‐hydroxy paclitaxel, 1′‐hydroxymidazolam, and 4‐hydroxy‐S‐mephenytoin were obtained from Toronto Research Chemicals.
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4

Synthesis and Characterization of Benzydamine Hydrogel

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Benzydamine hydrochloride (Mw 345.87 g/mol), sodium caseinate (from bovine milk) and CaCl2·2H2O (Mw 147.01 g/mol) were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other reagents were of analytical grade.
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5

Purification and Characterization of Enzymes

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2-Mercaptoethanol, Acetonitrile, Ampicillin sodium salt, Benzydamine hydrochloride, Benzydamine N-oxide hydrogen maleate, Bradford Reagent, d(+)-Glucose, Flavin adenine dinucleotide disodium salt hydrate (FAD), HisTrap HP columns, IGEPAL CA-630, Imidazole, Kanamycin, Nickel(II) sulfate hexahydrate, Lysozyme from hen egg white, Phenylmethylsulfonyl fluoride, and potassium phosphate were purchased from Sigma-Aldrich. NADPH tetra(cyclohexammoninum) and β-NADP sodium salt were purchased from Carbosynth. Isopropyl-beta-d-thiogalactopyranoside (IPTG) was purchased from BIOSYNTH. TRIS Base was purchased from Fisher Molecular Biology. Yeast Extract and Tryptone were purchased from Fisher Bioreagents. PageRuler Unstained Protein Ladder was purchased from Thermo Fisher Scientific Baltics UAB.
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6

Synthesis and Characterization of κ-Carrageenan Hydrogel

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Kappa-carrageenan (κ-CA), phosphorus pentasulfide (P4S10, 99%), tetramethylene sulfone (sulfolane, 99%), dimethyl sulfoxide-d6 (DMSO-d6, 99.9%), benzydamine hydrochloride, 5,5′-dithiobis(2-nitrobenzoic acid) (Ellman’s reagent), and sodium borohydride (NaBH4, ≥98%), dibutyltin dilaurate (DBTDL) and fluorescein isothiocyanate isomer 1 (FITC, ≥90%) were purchased from Sigma-Aldrich, Wien, Austria. Dialysis tubes of ZelluTrans from Carl Roth, Osterreich, Austria, with MWCO 3500 Da, were used. All the used chemicals were received from commercial sources and used without purification. Porcine small intestine was a donation from a slaughterhouse in Innsbruck.
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7

Quantitative Analysis of CBD and 7-OH-CBD Metabolism

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CBD and 7-OH-CBD were purchased
from Cayman Chemicals (Ann Arbor, MI) or Sigma-Aldrich (St. Louis,
MO). Pooled human liver microsomes (HLM) [n = 50,
mixed-gender (21 female and 29 male), race (42 Caucasian, 4 Hispanic,
2 African American, and 2 Asian), and age (5–77 years old)]
were obtained from Sekisui Xenotech, LLC (Lenexa, Kansas). NADPH-regenerating
system (1.3 mM NADP, 3.3 mM glucose 6-phosphate, and 0.4 U/mL glucose
6-phosphate dehydrogenase) was obtained from Corning (Bedford, MA).
Nicotine, norNicotine, HPBA, NOX, Nic-Gluc, cotinine, COX, Cot-Gluc,
3HC, and 3HC glucuronide (3HC-Gluc) standards were purchased from
Sigma-Aldrich. Fluconazole, benzydamine hydrochloride, clopidogrel,
trifluoperazine, diclofenac, amitriptyline, and tranylcypromine were
also purchased from Sigma-Aldrich. Optima grade methanol, acetonitrile,
and formic acid were obtained from Fisher Scientific (Waltham, MA).
Ultra-low-binding microcentrifuge tubes, Dulbecco’s modified
Eagle’s medium, Dulbecco’s phosphate-buffered saline,
UDP glucuronic acid (UDPGA), alamethicin, MgCl2, and Geneticin
(G418) were purchased from VWR (Radnor, PA). BCA protein assays were
purchased from Pierce (Rockford, IL); premium-grade fetal bovine serum
(FBS) was purchased from Seradigm (Radnor, PA), and ChromatoPur bovine
serum albumin (BSA) was purchased from MB Biomedicals (Santa Ana,
CA).
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8

Characterization of Drug Metabolism Assays

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Uridine 59-diphosphoglucuronic acid trisodium salt (UDPGA), NADPH, alamethicin from Trichoderma viride, adenosine 3-phosphate 5-phosphosulfate, diclofenac sodium salt, midazolam hydrochloride, felodipine, S-(+)-mephenytoin, raloxifene hydrochloride, lovastatin, nisoldipine, nifedipine, methadone, quinidine, testosterone, benzydamine hydrochloride, cisapride, cyclosporin, terbutaline, sildenafil, verapamil, trazodone, atorvastatin, simvastatin, buspirone, rifabutin, saquinavir, terfenadine, zolpidem, repaglinide, indinavir, alprazolam, carbazeran, enalapril, ramipril , and dabigatran etexilate were purchased from Sigma-Aldrich (St. Louis, MO). NADPH-regenerating system containing NADP + , glucose-6phosphate, and glucose-6-phosphate dehydrogenase was purchased from Corning (Wiesbaden, Germany). Organic solvents were of LC-mass spectrometry (MS) or higher quality grade and were acquired from VWR International (Radnor, PA) or Fisher Scientific UK Ltd (Loughborough, UK). Purified water was obtained from Milli-Q Integral 5 Water Purification System (Merck KGaA, Darmstadt, Germany). Phosphate buffer, HQM Hepatocyte/Enterocyte Incubation Medium, and Co-Factor N for MetMax were purchased from In Vitro ADMET Laboratories (Columbia, MD).
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9

Acquisition of Enzyme Substrates

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Dextrorphan tartrate, diclofenac sodium salt, 4-hydroxydiclofenac, S-mephenytoin, 4-hydroxyquinoline, paclitaxel, and testosterone were purchased from Cayman Chemical (Ann Arbor, MI). 7-Hydroxycoumarin was purchased from Chem Service (West Chester, PA). Benzydamine N-oxide, 7-Hydroxycoumarin sulfate potassium salt, kynuramine hydrobromide, and N-acetyl sulfamethazine were obtained from Santa Cruz Biotechnology (Dallas, TX). 4-Acetamidobenzoic acid, p-acetamidophenyl b-D-glucuronide sodium salt, 4-aminobenzoic acid, benzydamine hydrochloride, chlorzoxazone, coumarin, dextromethorphan hydrobromide, 6b-hydroxytestosterone, 7-Hydroxycoumarin b-D-glucuronide sodium salt, 7-ethoxycoumarin, paracetamol sulfate potassium, phenacetin, and sulfamethazine were purchased from Sigma-Aldrich (St. Louis, MO). Carbazeran, 4-hydroxyCarbazeran, 6-hydroxychlorzoxazone, 6a-hydroxypaclitaxel, acetaminophen glutathione disodium salt, midazolam, 19-hydroxymidazolam, and 4-hydroxy-S-mephenytoin were obtained from Toronto Research Chemicals (Toronto, ON, Canada). All other drug-metabolizing enzyme substrates were obtained from Sigma-Aldrich.
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