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8 protocols using hpβcd

1

Formulation and Packaging of Mouthwash

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RV-HPβCD complexation has already been described [18 (link),19 (link)]. The mouthwash was obtained as follows: RV (0.318% w/v, purchased from Fagron, Bologna, Italy) and HPβCD (0.318% w/v, purchased from Roquette, Lestrem, France) were mixed with deionized water in a clean and sterile recipient by means of a dissolver operated to shake at low speed without swirling for 5 h.
Then, a strawberry liquid aroma (Nyl Laboratories, Monterotondo, Italy) was added in an amount (weight/volume) of 0.1%, together with deionized water to reach 100% w/v. The choice to use the strawberry aroma was based on the results obtained in a preliminary study where a sample of children preferred it over the flavors tested (data not shown) and on the findings from previous published reports [22 ,23 (link)]. The obtained suspension was filtered and then packaged in a bottle provided with a spray solubilizer. All described procedures were performed by Logidex (Turin, Italy).
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2

Oleuropein-Cyclodextrin Formulation Development

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The materials used in this study were oleuropein (OLE; Sigma-Aldrich, St. Louis, MO, USA); hydroxypropyl-β-cyclodextrin parenteral grade (HP-β-CD; Kleptose, Roquette Freres, Lestrem, France); phosphatidylcholine (Pho; Lipoid® S 100; Lipoid, GmbH, Ludwigshafen, Germany); cholesterol (Chol; Sigma-Aldrich, St. Louis, MO, USA); Krebs–Ringer buffer solution (KRB, pH 7.4), variant without NaCl, with the following composition: 1.84 g/L D-glucose, 0.0468 g/L MgCl2, 0.34 g/L KCl, 0.1 g/L NaH2PO4, 0.18 g/L Na2HPO4; cell proliferation reagent WST-1 (Roche Diagnostic, Monza, Italy).
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3

Detailed Characterization of Ophthalmic Formulations

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DXMa was purchased from LA COOPER (Melun, France). Hydroxypropyl-γ-cyclodextrin (HPγCD, W8HP, DS = 0.6, and Mw = 1576 Da) was a kind gift from ASHLAND (Schaffhausen, Switzerland) and hydroxypropyl-β-cyclodextrin (HPβCD, KLEPTOSE DS = 0.63 and Mw = 1391 Da) was obtained from ROQUETTE (Lestrem, France). CELLUVISC® (sodium carboxymethylcellulose) and VISMED® (sodium hyaluronate) are marketed gels used for the treatment of dry eye syndrome. DEXAFREE® (DXM sodium phosphate 1% solution eye drops), MAXIDEX® (DXM 0.1% suspension eye drops) and BSS® (Alcon Laboratories, Rueil-Malmaison, France) are human authorized ocular medicines. Normal human primary corneal epithelial cells (ATCC PCS 700-010), medium (ATCC PCS-700-030), growth kit (ATCC PCS-700-040), PBS (ATCC 30-2200), trypsin EDTA (ATCC PCS-999-003 and 005), and antibiotics (gentamicin, streptomycin, and amphotericin BATC PCS-999-002) were obtained from LGC standard - ATCC® (Molsheim, France). Thioglycollate with resazurine medium and Tryptic soy broth were obtained from BIOMERIEUX (Craponne, France). ALAMARBLUE® was purchased from BIO-RAD (Marnes-la-Coquette, France) and DMSO from SIGMA-ALDRICH (Lyon, France). Purified water was prepared by DIRECT-Q®3UV water purifier (MILLIPORE, Molsheim, France). All other solvents and chemicals were of HPLC and analytical grade, respectively.
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4

Synthesis and Characterization of AFI

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AF was purchased from Euroclone S.p.a. (Milan, Italy). Hydroxypropyl-β–cyclodextrin (HPβ–CD, average molecular weight 1399, MS 0.65) was a generous gift of Roquette Pharma (Alessandria, Italy). AFI was synthesized according to the procedure reported in [3 (link)].
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5

Cannabidiol Quantification in Piglet Plasma

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Cannabidiol powder was purchased from THC Pharm GmHB (Frankfurt am Main, Germany). Cannabidiol-d3, 7-COOH-cannabidiol and 7-COOH-cannabidiol-d3 methanol solutions were purchased from LGC Ltd. (Teddington, UK) and Cerilliant Corporation (Round Rock, TX, USA), and stored at −20 °C. Piglet plasma was collected and purchased from PigforLife (Marloie, Belgium). Methanol and water were purchased from J.T. Baker by Filter Service (Eupen, Belgium), ethyl acetate was purchased from VWR Chemicals (Leuven, Belgium) and n-hexane 95% was purchased from Macron Fine Chemicals (VWR, Leuven, Belgium). All solvents were LCMS-grade. Sodium hydroxide and glacial anhydrous acetic acid 100% were purchased from Merck (Darmstadt, Germany); ammonium bicarbonate and ammonia 25% were obtained from Sigma-Aldrich (Saint Louis, MO, USA) and Fisher Chemicals (Hampton, NH, USA), respectively. HP-β-CD (Kleptose®, substitution degree of 0.62; Mw 1470 g/mol) was purchased from Roquette (Lestrem, France).
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6

Amiodarone-Cyclodextrin Formulation for Toxicity Study

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Amiodarone (AMD) (Mw = 645.32 Da) of 99.85% purity have been obtained from Zhejiang Sanmen Hengkang Pharmaceutical Co. Ltd., China. HP-β-CD of 99.70% purity have been obtained from Roquette, France. Polyoxyethylene (Hodge and Sterner, 1949 (link)) sorbitan monooleate (polysorbate 80, Tween 80) have been obtained from Sigma-Aldrich, Inc. Kollidon® SR (KOL), a mixture constituted from 80% poly(vinyl acetate) and 20% polyvinylpyrrolidone (povidone), and chitosan (CHT) were purchased from BASF, Germany.
AMD/HP-β-CD contains AMD:cyclodextrin in ratio 1:1. The substances were administered in constant volume (0.1 ml/10 g b.w.) as freshly prepared suspension with distilled water and 1% Tween-80. The solutions for acute toxicity study were prepared using the same weight of powder of AMD, respectively of AMD/HP-β-CD, Fc and F10 powders.
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7

Dexamethasone Sodium Phosphate Hydrogel

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Acrylic acid (AA), poly(vinyl pyrrolidone) K30 (PVP), and ammonium persulphate (APS) were procured from Daejung chemicals & metals Co, Ltd., Nakdong-daero, Sasang-gu, Busan, Korea. Hydroxylpropyl β cyclodextrin (HP-β-CD) was received from Roquette France as a gift sample. Ethanol and N, as well as N-methylene-bis-acrylamide (MBA) were acquired from Merck and Alfa Aesar, respectively. Remington Pharmaceutical Lahore, Pakistan gifted the dexamethasone sodium phosphate (DSP). The rest of the reagents employed were of analytical grade and used as received.
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8

Formulation and Characterization of DXM Eye Drops

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DXMa was purchased from LA COOPER (Melun, France). Hydroxypropyl-γ-cyclodextrin (HPγCD, W8HP, DS = 0.6 and Mw = 1576 Da) was a kind gift from ASHLAND (Schaffhausen, Switzerland) and Hydroxypropyl-β-cyclodextrin (HPβCD, KLEPTOSE DS = 0.63 and Mw = 1391 Da) was obtained from ROQUETTE (Lestrem, France). CELLUVISC® (sodium carboxymethylcellulose), GEL LARMES® (Carbopol 974P) and VISMED® (sodium hyaluronate) are marketed gels used for the treatment of dry eye syndrome. DEXAFREE® (DXM sodium phosphate 1% solution eye drops), MAXIDEX® (DXM 0.1% suspension eye drops) are human authorized ocular medicines. Methanol (HiPerSolvCHROMANORM for HPLC grade) was purchased from BDH, PROLABO (Leuven, Belgium. Purified water was prepared by DIRECT-Q® 3UV water purifier (MILLIPORE, Molsheim, France). All other solvents and chemicals were of HPLC and analytical grade, respectively.
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