Limonoids and Flavonoids from the Flowers of Azadirachta indica var. siamensis,and Their Melanogenesis‐Inhibitory and Cytotoxic Activities

A new limonoid, 7‐O‐acetyl‐7‐O‐debenzoyl‐22‐hydroxy‐21‐methoxylimocinin ( 2 ), and two new flavonoids, 3′‐(3‐hydroxy‐3‐methylbutyl)naringenin ( 7 ) and 4′‐O‐methyllespedezaflavanone C ( 9 ), along with nine known compounds, including two limonoids, 1 and 3 , and seven flavonoids, 4 – 6, 8 , and 10 – 12 , were isolated from a MeOH extract of the flowers of Azadirachta indica A.Juss. var. siamensis Valeton (Siamese neem tree; Meliaceae). The structures of new compounds were elucidated on the basis of extensive spectroscopic analysis and comparison with literature data. All of these compounds were evaluated for their melanogenesis‐inhibitory activities in B16 melanoma cells induced with α‐melanocyte‐stimulating hormone (α‐MSH). Compound 2 (16.9% melanin content at 30 μM ), 6‐deacetylnimbin ( 3 ; 49.6% melanin content at 100 μM ), and kaempferide ( 10 ; 41.7% melanin content at 10 μM ) exhibited inhibitory effects with no, or almost no, toxicity to the cells (81.0–111.7% cell viability). In addition, evaluation of their cytotoxic activities against HL60, A549, AZ521, and SK‐BR‐3 human cancer cell lines, isoazadironolide ( 1 ), 4′‐O‐methyl‐8‐prenylnaringenin ( 5 ), euchrestaflavanone A ( 8 ), 9 , and 3‐methoxy‐3′‐prenylnaringenin ( 12 ) revealed potent cytotoxicities against one or more cell lines with IC₅₀ values in the range of 4.5–9.9 μM .     

Biotransformation of cortexolone to hydrocortisone by molds using a rapid color development assay

Capabilities of 22 molds were assessed for 11beta-hydroxylation of cortexolone (Reichstein's compound S) to hydrocortisone. The biotransformation capability was compared for solid-state and submerged monocultures of the molds under otherwise identical conditions. A novel rapid color development assay and thin layer chromatography were used to qualitatively establish the ability of the fungi to convert cortexolone to hydrocortisone. These assays were validated and supplemented with data from high performance liquid chromatography to obtain quantitative information on the biotransformation. Nearly all the fungi consumed a significant fraction of the cortexolone fed, but only four (i.e. two isolates of Cunninghamella blakesleeana, C. echinulata and Curvularia lunata) yielded measurable quantities of hydrocortisone. Submerged cultures generally gave significantly greater yield of hydrocortisone compared to equivalent solid-state cultures.     

Effect of histamine and antihistamines on interleukin-1 production by human monocytes

This study was carried out on the effect of histamine hydrochloride and its antagonists on the production of interleukin-1 (IL-1) by lipopolysaccharide (LPS)-stimulated adherent human monocytes (AHM) from normal healthy blood donors. IL-1 activity was evaluated by incorporation of [3H]-thymidine in mouse thymocytes in samples of 1:3 dilution. The result indicated that histamine hydrochloride significantly suppressed IL-1 production by AHM at 10(-3) M and 10(-10) M in 14 donors with maximal suppression observed at 10(-3) M. A 1-hr incubation with histamine hydrochloride (10(-3) M) before addition of LPS was found to be appropriate. Cimetidine, an H2-antagonist at 10(-3) M, 10(-5) M, and 10(-7) M significantly inhibited the effect of histamine hydrochloride (10(-3) M) and gave maximum inhibition at 10(-5) M, whereas chlorpheniramine maleate, and H1-antagonist had no significant inhibitory effect at the concentrations studied (10(-4) M, 10(-5) M, and 10(-7) M). Histamine hydrochloride (10(-3) M) added alone had no significant suppressive effect, while cimetidine (10(-5) M) alone had a significant stimulatory effect on IL-1 production by AHM.