Product. For the mono-(O-acyl) derivatives (C5, C10 and C13) melting

Product. For the mono-(O-acyl) derivatives (C5, C10 and C13) melting point was decrease than GCV. When for the di-(O-acyl) derivatives (C5, C10 and C13) the melting points were additional reduce than mono-(O-acyl) derivatives (C5, C10 and C13) (Table 1). These outcomes indicate that raise in alkyl group conjugation lowered the melting points. Partition coefficient on the prodrugs was calculated with the aid of ACD labs software. Benefits indicate that ascending carbon chain length within the diesters long lipid conjugation enhanced the lipophilicity from the prodrugs. There was a large boost in calculated octanol/ water partition coefficient (logP) with conjugation of tridecanoic acid to both the hydroxyl groups of GCV (Table 1). These results recommend that the conjugation of lipid chain to GCV drastically enhances lipophilicity which may aid in improving GCV bioavailability together with slow release of GCV. Mass spectrometry All of the extended chain lipid GCV prodrugs and GCV were subjected to molecular weight analysis in positive mode with mass spectroscopy. Evaluation revealed GCV and all its derivatives as proton adduct [M+1]+except C13-mono-(O-acyl) derivative which wasNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptAdv Ophthalmol Vis Syst. Author manuscript; readily available in PMC 2014 October 30.Cholkar et al.Pageidentified as a sodium adduct [M+Na]+ (Table 1). Figure 1 shows the broad mass spectra for C13-mono-(O-acyl) GCV derivative (one hundred ng mL-1) having a mass of 452.4 Dalton and an intensity of 7.four 106 cps. All theoretically calculated masses for long chain lipid derivatives have been in agreement with mass spectroscopy outcomes (Table 1), indicating that all compounds were steady. Nuclear magnetic resonance The proton NMR spectral data and assignments for mono- and di-(O-acyl) derivatives were calculated in comparison for the parent compound, GCV (Table 2) [23]. Figures 2a and 2b shows the proton NMR for GCV and GCV-C5 conjugated lipid prodrugs. As is often seen inside the spectra for GCV prodrug (Figure 2a), we do not observe any alkyl resonance peaks downstream i.e. in between 0 ppm and 1.9 ppm. But, GCV-C5 exhibits the proton peaks corresponding to lipid chain conjugated to hydroxyl group of GCV (Figure 2b) which on integration determined the corresponded to proton quantity within the lipid chain. Related proton NMR spectra were collected for other prodrugs (GCV-C10 and GCV-13). Chemical shifts inside the resonance peaks for the lipid derivatives have been evident relative to GCV. Conjugation of aliphatic carbon chain (C5, C10 and C13) deshielded proton NMR chemical shifts of carbon (CH2OCO) to which these groups are conjugated as well as the -situated (OCH) protons.Biotin-azide Formula All other resonance peaks remained similar to that of GCV.Nikkomycin Z supplier Spectra had been comparable for mono- and di-(O-acyl) derivatives of GCV however the total number of protons calculated was double in number for di-(O-acyl) GCV lipid derivatives.PMID:24293312 13C NMR information for all prepared GCV lipid conjugates is presented in Table two. The ester conjugated carbon (RCOO) resonance peak was evident in all of the lipid prodrugs except the parent GCV molecule (Table two). These outcomes indicate that the extended carbon chain is conjugated to hydroxyl group of GCV. Cell culture To evaluate the cytotoxic effects of lengthy chain lipid GCV prodrugs MTS assay have been performed on ARPE-19 cells for 24 h. % viable cells were compared with of negative handle (culture medium) (Figure three). DMSO (ten ) served as good handle and decreased the % c.