Mp. 1H NMR and 13C NMR spectra had been recorded on a
Mp. 1H NMR and 13C NMR spectra had been recorded on a Bruker Avance DPX500 spectrometer with operating Epiregulin, Human frequencies of 500 and 125 MHz. All 13C NMR spectra were proton decoupled and all spectra have been obtained in deuterated chloroform (CDCl3). BMP-7 Protein Formulation Melting points were performed in triplicate using Gallenkamp melting point apparatus (Loughborough, UK) and were not corrected. High and low resolution MS (HRMS and LRMS) have been performed by the EPSRC National Mass Spectrometry Service, Swansea University, UK, working with the stated ionisation system. Elemental analyses, performed by Medac Ltd. (Surrey, UK) werePharmaceutics 2013,employed to confirm compound purity (95 ). Calculated logP (ClogP) values had been determined employing ChemDraw Ultra ten.0, CambridgeSoft, Cambridge, United states. 2.1.1. Process 1: Acid Chloride Synthesis Carboxylic acid derivatives of 4 or five had been dissolved in dry tetrahydrofuran (wv 75 mg10 mL) and cooled to 0 under nitrogen. Thionyl chloride (five equiv.) was added gradually with stirring followed by three drops of dimethyl formamide. The mixture was permitted to warm to ambient temperature and was stirred overnight. Solvents have been removed under vacuum as well as the acid chloride solution was used straight away with no additional purification. 2.1.two. Strategy 2: Dithranol Di-Ester Co-Drug Synthesis 1 (500 mg, 1 equiv.) was dissolved in dry tetrahydrofuran (30 mL) and cooled with dry iceacetone for 5 minutes with constant agitation. Pyridine (0.27 mL, 1.five equiv.) was added dropwise beneath nitrogen. The suitable acid chloride (two equiv.) was dissolved in dry tetrahydrofuran (2 mL), cooled (dry iceacetone) for five min, then added slowly into the mixture. The reaction was permitted to return to room temperature gradually and stirred overnight at space temperature. 1 mole per liter HCl (50 mL) was added as well as the volatiles have been removed by rotary evaporation. The mixture was extracted with dichloromethane (2 30 mL). The combined organic phases had been washed with saturated NaHCO3 answer (30 mL), dried more than MgSO4, and purified applying flash column chromatography applying dichloromethane 100 by means of to dichloromethane:ethyl acetate:petrol 13:1:6 as eluent. two.1.three. System three: Dithranol Mono-Ester Co-Drug Synthesis 1 (400 mg, 1 equiv.) was dissolved in 10 mL anhydrous hexamethylphoramide (HMPA) and chilled to 0 below nitrogen. The suitable acid chloride (1 equiv.) was dissolved in dry HMPA (3 mL), cooled to 0 , and was added in dropwise fashion for the dithranol solution. The mixture was slowly warmed to ambient temperature and permitted to stir for five h. The mixture was poured into 300 mL water and extracted with dichloromethane (three 40 mL). The combined organic phases were washed with water (3 100 mL) followed by saturated NaHCO3 answer (one hundred mL). The organic phase was further washed with water (two one hundred mL) and dried over MgSO4, prior to purification employing flash column chromatography. 2.1.4. Dithranol Dimer Synthesis (three) Prepared according to a published procedure [24], 1 (1 g) was dissolved in boiling acetic acid (one hundred mL), degassed and shielded from light. ten FeCl3 in acetic acid (12 mL) was added slowly. Water (five mL) was added plus the reaction item crystallized more than a period of several hours at area temperature. Re-crystallisation using acetic acid afforded the preferred product as a green powder (602 mg, 30 ). mp 24647 ; 1H NMR (CDCl3) 4.61 (s, 2H), 6.40 (d, 4H, J = 7.four Hz), 6.93 (d, 4H, J = eight.3 Hz), 7.41 (t, 4H, J = eight.0, 7.eight Hz), 11.73 (s, 4H, OH). 13C NMR (CDCl3) 56.three, 116.7, 11.