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3 protocols using glycerol monomethacrylate

1

Synthesis of Poly(2-hydroxyethyl acrylate)

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The poly(ethylene glycol) methyl ether acrylate (PEGMEA480, MW 480), poly(ethylene glycol) dimethacrylate (PEGDMA550, Mw 550), sodium hydroxide (NaOH), 3-(acryloyloxy)-2-hydroxypropyl methacrylate, acetic acid, Sudan I, phenylbis(2,4,6-trimethyl benzoyl) phosphine oxide (BAPO), and phosphate-buffered saline (PBS, pH 7,4) were supplied by Merck and used as received without further purification.
The Jeffamine ED-600 (supplied by Huntsman) and glycerol monomethacrylate (GMM, supplied by Polysciences) were used as received.
2-hydroxy ethyl acrylate (HEA, supplied by TCI), azo-bis-isobutirenitrile (AIBN, supplied by Glentham) and dioxane (supplied by Panreac) were used as received.
The HCL was prepared as described previously from 3-(acryloyloxy)-2-hydroxypropyl methacrylate and Jeffamine using a catalytic quantity of acetic acid [20 ].
The synthesis of poly-HEA (pHEA) was carried out by thermal polymerization in solution. 665 mg of HEA were introduced into a vial, with a stirrer inside, and dissolved in 10 mL of dioxane. Next, 25 mg of AIBN were added. After that, the solution was left stirring for about 5–10 min for its complete homogenization and then nitrogen was bubbled for 5 min. Finally, the polymerization was allowed to proceed overnight in an oven at 60 °C. The next day, upon cooling to 5 °C separation into two phases was observed, which allows the dioxane to be easily removed.
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2

Synthesis and Characterization of Polymeric Materials

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4-Cyano-4-(thiobenzoylthio) pentanoic acid (CPAD, 97%), 4,4′-azobis(4-cyanopentanoic acid) (ACVA, ≥98%), poly(ethylene glycol) methyl ether methacrylate (PEGMA, Mn 500 g·mol−1), N,N-dimethyl acrylamide (DMA, 99%), 2,2′-azobis(2-methylpropionitril) (AIBN, 98%), and 1,3,5-trioxane (≥99%) were obtained from Aldrich (Saint Louis, MO, USA) and used as received. 2-(Dodecylthiocarbonothioylthio)-2-methylpropionic acid (DTMPA, 97%) was purchased from Strem Chemicals (Boston, MA, USA) and utilized as received. Glycerol mono-methacrylate (GMMA, 95%) was acquired from Polysciences, Inc. (Warrington, PA, USA), and acrylic acid (AA, 99.5%) was obtained from Alfa Aesar (Tewksbury, MA, USA), and used as received. All monomers were purified by stirring in the presence of inhibitor-remover beads (for hydroquinone and monomethyl ether hydroquinone, Aldrich, Saint Louis, MO, USA). 1,4-Dioxane (99.5%) was purchased from Acros Organics (Geel, Belgium), and used as received. Ethanol was provided by the solvent purification system MB-SPS-800 (MBraun, Stratham, NH, USA) and stored under argon. Float-A-Lyzer®® G2 dialysis tubes (0.5 to 1 kDa, dmembrane = 10 mm, volume = 10 mL) were acquired from Repligen (former Spectrum Labs, Waltham, MA, USA) and used for the dialysis of the polymer solutions.
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3

Synthesis and Characterization of Tenofovir Alafenamide Polymer Conjugates

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Materials were purchased from Sigma-Aldrich unless otherwise specified. All solvents were Fisher HPLC grade. Chain transfer agents, 4-(((2-carboxyethyl)thiocarbonothioyl)thio)-4cyanopentanoic acid (CCC) and 4-cyano-4-((ethylsufanylthiocarbonyl)-sulfanyl)pentanoic acid (ECT) were purchased from Boron Molecular and OMM Scientific (Texas, USA), respectively.
Glycerol monomethacrylate was purchased from Polysciences and purified using basic alumina before polymerization. 2,2'-Azobis(4-methoxy-2,4-dimethylvaleronitrile) (V-70) was purchased from FUJIFILM Wako Pure Chemical Corporation, USA and used without further purification.
Fasudil hydrochloride was purchased from eNovation Chemicals (Green Brook, NJ, USA). Spectra/Por regenerated cellulose dialysis membranes were purchased from Spectrum Laboratories (Houston, TX, USA). Rhodamine B methacrylate (RhMA) was synthesized as described previously. [43] (link) Tenofovir alafenamide (TAF) fumarate (1:1 salt) was obtained from MedKoo Biosciences, Inc. Analytical standards: tenofovir (TFV) and tenofovir diphosphate (TFV-DP) were obtained from Cayman Chemical and Santa Cruz Biotech, respectively. Isotopic internal standards: TAF-d5 fumarate, TFV-d6 and TFV-DP 13 C5 were obtained from Toronto Research Chemicals and Moravek, respectively. HPLC grade methanol and water were purchased from VWR international Ltd.
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