An innovative strategy dedicated to quality-by-design (QbD) principles has been comprehensively applied in the set-up of a vortex aided salting-out-assisted liquid–liquid microextraction (VA-SALLME) combined with a core–shell chromatographic method for the simultaneous analysis of two anti-glaucoma drugs, dorzolamide hydrochloride (DOR) and timolol maleate (TIM), in rabbit plasma. Each step of the QbD workflow has been implemented, starting from using the design of experiments with a Plackett–Burman design for screening and a Box–Behnken design for response surface methodology up to using Monte Carlo simulations for error propagation. The optimal chromatographic conditions were: a core–shell Kinetex XB C18 column with ACN: 45.56 mmol L-1 phosphate buffer at pH 3.76 (18.17 : 81.83%, v/v) as the mobile phase at a 1 mL min-1 flow rate, using hydrochlorothiazide as an internal standard and detection at 254 and 295 nm for DOR and TIM, respectively. Incorporating QbD principles in the method development allowed the visualization of two definite design spaces for the VA-SALLME and the HPLC methods, providing assurance of the quality in accordance with ICH guideline Q8(R2). Linearity was found over the concentration ranges of 0.9–50 and 1.5–50 ng mL-1 with quantitation limits of 0.87 and 1.40 ng mL-1 for DOR and TIM, respectively. As a consequence of the obtained unprecedented chromatographic separation and sensitivity, it was possible to carry out simultaneous pharmacokinetic studies of the two anti-glaucoma drugs after their single instillation.

An innovative strategy dedicated to quality-by-design (QbD) principles has been comprehensively applied in the set-up of a vortex aided salting-out-assisted liquid–liquid microextraction (VA-SALLME) combined with a core–shell chromatographic method for the simultaneous analysis of two anti-glaucoma drugs, dorzolamide hydrochloride (DOR) and timolol maleate (TIM), in rabbit plasma. Each step of the QbD workflow has been implemented, starting from using the design of experiments with a Plackett–Burman design for screening and a Box–Behnken design for response surface methodology up to using Monte Carlo simulations for error propagation. The optimal chromatographic conditions were: a core–shell Kinetex XB C18 column with ACN: 45.56 mmol L-1 phosphate buffer at pH 3.76 (18.17 : 81.83%, v/v) as the mobile phase at a 1 mL min-1 flow rate, using hydrochlorothiazide as an internal standard and detection at 254 and 295 nm for DOR and TIM, respectively. Incorporating QbD principles in the method development allowed the visualization of two definite design spaces for the VA-SALLME and the HPLC methods, providing assurance of the quality in accordance with ICH guideline Q8(R2). Linearity was found over the concentration ranges of 0.9–50 and 1.5–50 ng mL-1 with quantitation limits of 0.87 and 1.40 ng mL-1 for DOR and TIM, respectively. As a consequence of the obtained unprecedented chromatographic separation and sensitivity, it was possible to carry out simultaneous pharmacokinetic studies of the two anti-glaucoma drugs after their single instillation.

The progestational potency and selectivity of synthetic steroidal agonists can be enhanced by even larger chemical moieties at 17a-position of the steroid backbones. Hereby a series 5a-c and 6a-c of novel 17a-[1-(substituted phenyl)-1,2,3-triazol-4-yl]-19-nortestosterone-17b-yl acetates were designed and synthesized using click chemistry approach searching progestogenic derivatives with potential anticancer activity. Compounds 5a,b and 6a,c have affected to different extents the three histopathological parameters considered for evaluation of their progestational activity. The compounds 5a,b and 6a,c showed modifications in rat uterus at 35.7e34.8 nM levels with privileged endometrial thickening effect and least change of uterine weight relative to NEA at 52.9 nM level. Up to 40 mg/kg dose compounds 5b and 6c were non-toxic. Molecular docking of the ligands in PR showed in the majority of cases a conformational fitting into the active site different from that of the reference steroid NEA. Compound 6b revealed about 46.4% growth inhibition of CNS cancer SNB-75 cell line, 56% growth inhibition of renal cancer A498 cell line and 56.7% growth inhibition of prostate cancer PC-3 cell line which was mediated by cell cycle arrest. Drugability of the screened compounds showed tolerated results after being challenged to diverse physicochemical parameters.

The progestational potency and selectivity of synthetic steroidal agonists can be enhanced by even larger chemical moieties at 17a-position of the steroid backbones. Hereby a series 5a-c and 6a-c of novel 17a-[1-(substituted phenyl)-1,2,3-triazol-4-yl]-19-nortestosterone-17b-yl acetates were designed and synthesized using click chemistry approach searching progestogenic derivatives with potential anticancer activity. Compounds 5a,b and 6a,c have affected to different extents the three histopathological parameters considered for evaluation of their progestational activity. The compounds 5a,b and 6a,c showed modifications in rat uterus at 35.7e34.8 nM levels with privileged endometrial thickening effect and least change of uterine weight relative to NEA at 52.9 nM level. Up to 40 mg/kg dose compounds 5b and 6c were non-toxic. Molecular docking of the ligands in PR showed in the majority of cases a conformational fitting into the active site different from that of the reference steroid NEA. Compound 6b revealed about 46.4% growth inhibition of CNS cancer SNB-75 cell line, 56% growth inhibition of renal cancer A498 cell line and 56.7% growth inhibition of prostate cancer PC-3 cell line which was mediated by cell cycle arrest. Drugability of the screened compounds showed tolerated results after being challenged to diverse physicochemical parameters.

The progestational potency and selectivity of synthetic steroidal agonists can be enhanced by even larger chemical moieties at 17a-position of the steroid backbones. Hereby a series 5a-c and 6a-c of novel 17a-[1-(substituted phenyl)-1,2,3-triazol-4-yl]-19-nortestosterone-17b-yl acetates were designed and synthesized using click chemistry approach searching progestogenic derivatives with potential anticancer activity. Compounds 5a,b and 6a,c have affected to different extents the three histopathological parameters considered for evaluation of their progestational activity. The compounds 5a,b and 6a,c showed modifications in rat uterus at 35.7e34.8 nM levels with privileged endometrial thickening effect and least change of uterine weight relative to NEA at 52.9 nM level. Up to 40 mg/kg dose compounds 5b and 6c were non-toxic. Molecular docking of the ligands in PR showed in the majority of cases a conformational fitting into the active site different from that of the reference steroid NEA. Compound 6b revealed about 46.4% growth inhibition of CNS cancer SNB-75 cell line, 56% growth inhibition of renal cancer A498 cell line and 56.7% growth inhibition of prostate cancer PC-3 cell line which was mediated by cell cycle arrest. Drugability of the screened compounds showed tolerated results after being challenged to diverse physicochemical parameters.

The progestational potency and selectivity of synthetic steroidal agonists can be enhanced by even larger chemical moieties at 17a-position of the steroid backbones. Hereby a series 5a-c and 6a-c of novel 17a-[1-(substituted phenyl)-1,2,3-triazol-4-yl]-19-nortestosterone-17b-yl acetates were designed and synthesized using click chemistry approach searching progestogenic derivatives with potential anticancer activity. Compounds 5a,b and 6a,c have affected to different extents the three histopathological parameters considered for evaluation of their progestational activity. The compounds 5a,b and 6a,c showed modifications in rat uterus at 35.7e34.8 nM levels with privileged endometrial thickening effect and least change of uterine weight relative to NEA at 52.9 nM level. Up to 40 mg/kg dose compounds 5b and 6c were non-toxic. Molecular docking of the ligands in PR showed in the majority of cases a conformational fitting into the active site different from that of the reference steroid NEA. Compound 6b revealed about 46.4% growth inhibition of CNS cancer SNB-75 cell line, 56% growth inhibition of renal cancer A498 cell line and 56.7% growth inhibition of prostate cancer PC-3 cell line which was mediated by cell cycle arrest. Drugability of the screened compounds showed tolerated results after being challenged to diverse physicochemical parameters.

A new cycloartane-type saponin with unusual hydroxylation at C-17 and a unique side chain, 9 (R), 19, 22 (S), 24 (R) bicyclolanost-3β, 12α, 16β, 17α tetrol-25-one 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (1) and two new monoterpenoid glucoindole alkaloids, 10-methoxy pumiloside (2) and the previously chemically synthesized, 10-methoxy strictosidine (3) along with other five known compounds, 7α-morroniside (4), 7-epiloganin (5), (7β)-7-O-methylmorroniside (6), 5(S)-5-carboxystrictisidine (7) and apigenin-7-Oneohesperidoside (8) were isolated from the aerial parts of Mussaenda luteola (Rubiaceae). The structural elucidation of the isolates was accomplished by extensive (1D and 2D NMR) spectroscopic data analysis and HR-ESI-MS. Compounds 4–8 were reported for the first time from the genus Mussaenda. Interestingly, this is the first report for the occurrence of the monoterpenoid glucoindole-type alkaloids in the genus which might be useful for the chemotaxonomic evaluation of the genus Mussaenda. All isolates were evaluated for their antiprotozoal activities. Compound 7 showed good antitrypanosomal activity with IC50 and IC90 values of 13.7 and 16.6 μM compared to IC50 and IC90 values of 13.06 and 28.99 μM for the positive control DFMO, difluoromethylornithine.

A new cycloartane-type saponin with unusual hydroxylation at C-17 and a unique side chain, 9 (R), 19, 22 (S), 24 (R) bicyclolanost-3β, 12α, 16β, 17α tetrol-25-one 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (1) and two new monoterpenoid glucoindole alkaloids, 10-methoxy pumiloside (2) and the previously chemically synthesized, 10-methoxy strictosidine (3) along with other five known compounds, 7α-morroniside (4), 7-epiloganin (5), (7β)-7-O-methylmorroniside (6), 5(S)-5-carboxystrictisidine (7) and apigenin-7-Oneohesperidoside (8) were isolated from the aerial parts of Mussaenda luteola (Rubiaceae). The structural elucidation of the isolates was accomplished by extensive (1D and 2D NMR) spectroscopic data analysis and HR-ESI-MS. Compounds 4–8 were reported for the first time from the genus Mussaenda. Interestingly, this is the first report for the occurrence of the monoterpenoid glucoindole-type alkaloids in the genus which might be useful for the chemotaxonomic evaluation of the genus Mussaenda. All isolates were evaluated for their antiprotozoal activities. Compound 7 showed good antitrypanosomal activity with IC50 and IC90 values of 13.7 and 16.6 μM compared to IC50 and IC90 values of 13.06 and 28.99 μM for the positive control DFMO, difluoromethylornithine.

A new cycloartane-type saponin with unusual hydroxylation at C-17 and a unique side chain, 9 (R), 19, 22 (S), 24 (R) bicyclolanost-3β, 12α, 16β, 17α tetrol-25-one 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (1) and two new monoterpenoid glucoindole alkaloids, 10-methoxy pumiloside (2) and the previously chemically synthesized, 10-methoxy strictosidine (3) along with other five known compounds, 7α-morroniside (4), 7-epiloganin (5), (7β)-7-O-methylmorroniside (6), 5(S)-5-carboxystrictisidine (7) and apigenin-7-Oneohesperidoside (8) were isolated from the aerial parts of Mussaenda luteola (Rubiaceae). The structural elucidation of the isolates was accomplished by extensive (1D and 2D NMR) spectroscopic data analysis and HR-ESI-MS. Compounds 4–8 were reported for the first time from the genus Mussaenda. Interestingly, this is the first report for the occurrence of the monoterpenoid glucoindole-type alkaloids in the genus which might be useful for the chemotaxonomic evaluation of the genus Mussaenda. All isolates were evaluated for their antiprotozoal activities. Compound 7 showed good antitrypanosomal activity with IC50 and IC90 values of 13.7 and 16.6 μM compared to IC50 and IC90 values of 13.06 and 28.99 μM for the positive control DFMO, difluoromethylornithine.

A new iridoid glucoside, 10-methoxy apodanthoside (1), and a new monoterpene glycoside, (3S,6S)-cis linalool-3,7-oxide O-β-ᴅ-glucopyranosyl-(1''→5')-β-ᴅ-xylofuranoside (2), were isolated from V. edulis (Rubiaceae), along with eighteen known compounds (3-20), including monoterpenes, iridoid glycosides, and a lignin, which were encountered for the first time in the genus Vangueria,. The structural elucidation of the isolates was based on the analysis of spectroscopic (1D and 2D NMR) and HR-ESI-MS data. Detailed stereochemical studies of 1 and related iridoid glucosides (compounds 3, 4 and 8) were made by matching the calculated ECD peaks with the experimental ones. All isolates were tested for their antiprotozoal, antifungal, and antiplasmodial activities. Compounds 9, 15 and 16 showed good trypanocidal activities against Trypanosoma brucei brucei with IC50 values of 8.18, 9.02 and 7.80 μg/mL, respectively and IC90 values of >10, >10 and 9.76 μg/mL, respectively. Compound 16 showed a moderate activity against Candida glabrata with an IC50 value of 8.66 μg/mL. Compound 20 showed a weak antiplasmodial activity against chloroquine-sensitive (D6) and resistant (W2) Plasmodium falciparum with IC50 values of 3.29 (SI, >1.4) and 4.53 (SI, >1) μg/mL, respectively