Three verrucosane diterpenoids,verrucosane triol and related compounds from the liverwort Mylia verrucosa

Three new diterpenoids of the verrucosane class were isolated as minor constituents of the methanol extract of the liverwort Mylia verrucosa and their structures shown to be 2β,9α,13β-trihydroxyverrucosane,9α-acetoxy-2β,13β-dihydroxyverrucosane and 2β,13β-dihydroxy-9-oxoverrucosane.     

The structure of mollugogenol-f,a new triterpenoid sapogenin from Mollugo hirta

The constitution of mollugogenol-F, a new triterpenoid sapogenin isolated from Mollugo hirta (family: Ficoidaceae), has been established as 3β,16β,22-trihydroxyisohopane.     

16β-Hydroxycholestanol als biogenetische vorstufe der steroidsapogenine

It has been shown that 5,6-³H,16β-hydroxycholestanol is used in the biogenesis of the steroid sapogenins, tigogenin and gitogenin, by plants of Digitalis lanata but not for the formation of tomatidine by Solanum lycopersicum.     

Improved synthesis of 17β-hydroxy-16α-iodo-wortmannin, 17β-hydroxy-16α-iodoPX866, and the [131I] analogue as useful PET tracers for PI3-kinase

An improved method for the synthesis of 17β-hydroxy-16α-iodo-wortmannin along with the first synthesis of 17β-hydroxy-16α-iodoPX866 and [¹³¹I] radiolabeled 17β-hydroxy-16α-[¹³¹I]iodo-wortmannin, as potential PET tracers for PI3K was also described. The differences between wortmannin and its iodo analogue were compared by covalently docking each structure to L833 in PI3K.     

Preparation of 2- and 3-substituted gibberellins A9 and A4 for bioassay

To test the effects of preventing enzymatic 2β- and 3β-hydroxylation on the biological activities of gibberellins, the preparation of the following compounds is described: 2β-methyl- and 2,2-dimethyl-gibberellins A₄ and A₉; 2α-fluoro-, 2β-fluoro- and 2β-methoxy-gibberellin A₉; and 3β-chloro-, 3β-fluoro-, 3β-methoxy- and 3-methylene A₉.     

Structure of inflexinol,a new cytotoxic diterpene from Rabdosia inflexa

From the leaves of Rabdosia inflexa a new cytotoxic diterpenoid, inflexinol was isolated, together with the known inflexin, and the structure was established as ent-kaur-16-en-15-one-1α,3α,6β,11β-tetraol 1,3-diacetate.     

PTP1B inhibitory activity of kaurane diterpenes isolated from Siegesbeckia glabrescens

Protein tyrosine phosphatase 1B (PTP1B) is considered as a therapeutic target for the treatment of diabetes and obesity. In our preliminary screening study, a MeOH extract of the aerial part of Siegesbeckia glabrescens was found to inhibit PTP1B activity at 30 μg/mL. Bioassay‐guided fractionation led to the isolation of two active diterpenes, ent-16βH,17-isobutyryloxy-kauran-19-oic acid (1) and ent-16βH,17-acetoxy-18-isobutyryloxy-kauran-19-oic acid (2), along with ent-16βH,17-hydroxy-kauran-19-oic acid (3). Compounds 1 and 2 inhibited the PTP1B activity with IC₅₀ values of 8.7 ± 0.9 and 30.6 ± 2.1 μM, respectively. Kinetic studies suggest that both 1 and 2 are non-competitive inhibitors of PTP1B. However, compound 3 substituted with a hydroxyl group at C-17 in kaurane-type showed no inhibitory effects towards PTP1B.     

Beckmann reactions of steroidal spirocyclic oximes derived from the 16beta,23:23,26-diepoxy-22-oxo moiety

The Beckmann rearrangement of the syn and anti isomers of the spirocyclic oxime derived from a 16β,23:23,26-diepoxy-5β-cholestan-22-one was studied. Whereas the anti isomer always follows the Beckmann fragmentation course, the syn isomer, depending on the reaction conditions, follows the normal Beckmann rearrangement course and/or the isomerization to the anti isomer followed by the fragmentation course.     

Carotenoids of Anacystis nidulans,structures of caloxanthin and nostoxanthin

Reinvestigation of the carotenoids of Anacystis nidulans has confirmed the occurrence of β,β-carotene (β-carotene), β,β-caroten-3-ol (cryptoxanthin), β,β-carotene-3,3′-diol (zeaxanthin) and 2R,3R,3′R-β,β-carotene-2,3,3′-triol (absolute configuration assigned in the present work). In addition the previously unknown 2R,3R,2′R,3′R-β,β-carotene-2,3,2′,3′-tetrol has been isolated. The triol and the tetrol are considered identical with caloxanthin and nostoxanthin, respectively, for which allenic structures have been suggested by others. The chirality of these compounds followed from CD and ¹H NMR considerations.     

A new triterpene lactone from Gymnocladus dioica

2β,23-Dihydroxyacacic acid lactone was isolated from Gymnocladus dioica as an artifact from 2β, 3β 16β, 21β, 23-pentahydroxyolean-12-ene-28-oic acid.     

Estrogen metabolism in nonhuman primates I. In vitro biosynthesis of estrogen glucosiduronates in rhesus monkey liver

Estrone glucosiduronate, 17β-estradiol-3-glucoslduronate, 17β-estradiol-17-glucosiduronate and estriol-16α-glucoslduronate have been biosynthesized in substantial yield by incubation of radioactive estrone, 17β-estradiol, estriol and uridlne diphosphoglucosiduronic acid with rhesus monkey liver homogenates. The metabolites were characterized by chromatography on Celite and DEAE-Sephadex, enzyme hydrolysis, derivative formation and crystallization to constant specific activity. The percent conversion to 17β-estradiol-17-glucosiduronate from 17β-estradlol ranged between 56–71%; from estrone, the conversion was 49–54%. Other metabolites formed from estradiol were estrone glucosiduronate(12–21%) and 17β-estradiol-3-glucosiduronate(5–12%). The same metabolites derived from estrone accounted for 18–28% and 10–14% respectively. After estriol incubation, more than 90% of the steroid was converted to estriol-16α-glucosiduronate with no detectable estriol-3-glucosiduronate. This report represents the first time that 17β-estradiol-17-glucosiduronate has been reported as a metabolite in the rhesus monkey and this is the only known species which forms 17β-estradiol-17-glucosiduronate as the predominant metabolite of either estrone or estradiol $\text{in vitro}$.Rhesus monkey liver is similar to the human and baboon in that it metabolizes estriol exclusively to estriol-16-glucosiduronate.     

A New Diterpene Glycoside in Aster ageratoides Turcz.

A new natural diterpene glycoside was isolated from Aster ageratoides Turcz. Its structure was elucidated by means of IR, MS (FAB, EI), NMR (1H-, 13C-, DEPT), 2D-NMR (1H-1H COSY, 13C-1 H COSY, J-resolved, CoLoC) spectra and chemical method as 16β, 17-dihydroxy- (—) -kauran- 19-oic acid-β-D-glucopyanosyl ester.     

Cycloartane glycosides from Astragalus campylosema Boiss. ssp. campylosema

Four cycloartane glycosides, 3-O-[α-l-arabinopyranosyl-(1 → 2)-β-d-xylopyranosyl]-3β,6α,16β,23α,25-pentahydroxy-20(R),24(S)-epoxycycloartane (1), 3-O-[α-l-arabinopyranosyl-(1 → 2)-β-d-xylopyranosyl]-16-O-hydroxyacetoxy-23-O-acetoxy-3β,6α,25-trihydroxy-20(R),24(S)-epoxycycloartane (2), 3-O-[α-l-arabinopyranosyl-(1 → 2)-β-d-xylopyranosyl]-3β,6α,23α,25-tetrahydroxy-20(R),24(R)-16β,24;20,24-diepoxycycloartane (3), 3-O-[α-l-arabinopyranosyl-(1 → 2)-β-d-xylopyranosyl]-25-O-β-d-glucopyranosyl-3β,6α,16β,25-tetrahydroxy-20(R),24(S)-epoxycycloartane (4), along with three known cycloartane glycosides were isolated from the MeOH extract of the roots of Astragalus campylosema ssp. campylosema. Their structures were established by the extensive use of 1D- and 2D-NMR experiments along with ESIMS and HRMS analysis. The occurrence of the hydroxyl function at position 23 (1-2) and of the ketalic function at C-24 (3) are very unusual findings in the cycloartane class.     

Triterpenoids of Salvia horminum,constitution of a new diol

Lupeol, lup-(20)29-ene-2α,3β-diol and a new triterpenic alcohol olean-(13)18-ene-2β,3β-diol were isolated from the petrol extract of air dried Salvia horminum and their structures were determined.     

Neutrophil CD16b crosslinking induces lipid raft-mediated activation of SHP-2 and affects cytokine expression and retarded neutrophil apoptosis

Two different types of FcRs for IgG are constitutively expressed on the surface of human neutrophils, namely, FcγRIIA (CD32a) and FcγRIIIB (CD16b). Unlike FcγRIIA, FcγRIIIb is GPI anchored to the cell membrane and its signal transduction is still ambiguous. To further understand the signal transduction of CD16b, we compared neutrophil cytokine expression and apoptosis by the cross-linking of CD32a and CD16b respectively. We found that both CD32a and CD16b crosslinking can activate neutrophils, but did not exactly share cytokine expression profiles. On the other hand, CD16b cross-linking retarded neutrophil apoptosis while CD32a promoted it. By interrupting the lipid raft with methyl-β-cyclodextrin (MβCD) and inhibiting the ITAM-SYK pathway with an SYK inhibitor (piceatannol), we found reduced apoptosis was at least partially mediated by lipid raft structure, but not the ITAM-SYK pathway. Additionally, CD16b but not CD32a cross-linking triggered SHP-2 phosphorylation and led to its translocation into lipid rafts. SHP-2 phosphorylation and translocation were inhibited by MβCD. Moreover, pre-inhibition of SHP-2 by a specific inhibitor (SHP099) converted IL-10 and SOCS3 expression level and promoted neutrophil apoptosis after CD16b crosslinking. In conclusion, these results, for the first time, collectively indicate that SHP-2 is activated by CD16b crosslinking in neutrophils and functions as a component of the raft-mediated signaling pathway.     

Nanoencapsulation of triterpene 3β,6β,16β-trihydroxylup-20(29)-ene from Combretum leprosum as strategy to improve its cytotoxicity against cancer cell lines

The pentacyclic triterpene 3β,6β,16β-tri-hydroxilup-20(29)-ene is a natural product produced by the Brazilian medicinal plant Combretum leprosum. Its cytotoxicity has been previously reported against breast cancer cell lines. The low water solubility of this natural product, that hampers its bioavailability, motivated the investigation of a new nanoparticle formulation containing the triterpene in order to improve its bioactivity. The triterpene was encapsulated in polycaprolactone (PCL) polymer by nanoprecipitation, producing homogenic nanoparticles with nanometer sizes (122.7 ± 2.06 nm), which were characterized by FT-IR, SEM imaging and DSC. The cytotoxicity (MTT method) of the nanoparticle containing the triterpene 1, besides the free natural product and the nanoparticle control (without 1), was assayed against three human tumor cell lines [human colon carcinoma line (HCT116), prostate (PC3) and glioblastoma (SNB19)] and the normal epithelial embryo kidney human cell line (Hek293T). The nanocarrier produced a significative effect in the cytotoxicity of the natural product in the nanoformulation (IC₅₀ 0.11–0.26 µg mL⁻¹) when compared with its free form (IC₅₀ 1.07–1.44 µg mL⁻¹). Additionally, higher selectivity of the triterpene to the tumor cells was found when it was encapsulated (SI 1.92–4.54) than in its free form (SI 0.42–0.56). In this case, the nanoencapsulated triterpene was more selective to PC3 (SI 3.33) and SNB19 (SI 4.54) tumor cells.     

Kolavane and kaurane diterpenes from the stem bark of Xylopia aethiopica

The stem bark of Xylopia aethiopica has yielded four diterpenes, two of them novel. Three of the diterpenes were identified as (?)-kaur-16-en-19-oic acid and its 7-oxo and 7β-hydroxy derivatives. The fourth was the novel kolavane derivative 2-oxo-kolav-3,13-dien-15-oic acid, a type of compound not previously recorded in the Annonaceae.     

IKKβ specifically binds to P16 and phosphorylates Ser8 of P16

In spite of its central roles in cell cycle progression, senescence, and aging, knowledge about the posttranslational regulation of P16 (also known as INK4A and MTS1) remains limited. While it has been reported that P16 could be phosphorylated at Ser7, Ser8, Ser140, and Ser152, the corresponding kinases have not been identified yet. Here we report that IKKβ, a primary kinase for IκBα phosphorylation, is involved in P16 phosphorylation. Immunoprecipitation and kinase assays showed that IKKβ specifically binds to P16 and phosphorylates P16 at Ser8 in WI38 cells. Biochemical characterization of phosphomimetic Ser → Glu P16 mutants demonstrated that phosphorylation at Ser8 of P16 brings about a significant loss of its cyclin-dependent kinase (CDK) 4-inhibitory activity while P16 retains structurally and functionally intact upon phosphorylation at Ser7, Ser140, and Ser152. Our results reveal the novel role of IKKβ in P16 phosphorylation and broaden our understanding of the regulation of P16.     

Synthesis and biological evaluation of α,β-unsaturated lactones as potent immunosuppressive agents

Compounds having α,β-unsaturated lactones display a variety of biological activities. Many research groups have tested both natural and unnatural α,β-unsaturated lactones for as-yet undiscovered biological properties. We synthesized α,β-unsaturated lactones with various substituents at the δ-position and studied their immunosuppressive effects, that is, the inhibition of Interleukin-2 (IL-2) production. Among the compounds synthesized, the benzofuran-substituted α,β-unsaturated lactone 4h showed the best inhibitory activity toward IL-2 production in Jurkat e6-1 T lymphocytes (IC(50)=66.9 nM) without cytotoxicity at 10 μM. The results indicated that 4h may be useful as a potent immunosuppressive agent, as well as in IL-2-related studies.     

β2-microglobulin is overexpressed in buccal cells of elderly and correlated with expression of p16 and inflammatory genes

β2M (Beta 2 microglobulin) is a small protein that is found in all nucleated cells, previous finding showed that its levels increased in the serum of the elderly. Buccal cell samples are none invasive approach for assessing the expression of target genes. There was rationality to assess the expression of β2M in buccal cells of people of a different group of ages. Indeed, the expression of β2M increased significantly with fold change 3.40, 4.80, 6.60^**, 8.20^*** and 12.04^*** for the group of age 18–25 years, 26–35 years, 36–45 years, 46–55 years, and 56–70 years respectively. The same observation was seen with markers of biological aging (p16^INK4a) with fold change 3.19, 3.90, 4.80*, 8.50^*** and 12.40^*** for the group of age 18–25 years, 26–35 years, 36–45 years, 46–55 years, and 56–70 years respectively. As expected, there was an increase in the inflammatory genes (IL-1 β and IL-6) expression in the elderly. Moreover, there was a direct significant correlation (r = 90, p < 0.001) between β2M expression and age (years), and the same direct significant correlation between p16^INK4a expression and age (years) was also seen (r = 90, p < 0.001). In addition, a direct correlation between β2M and p16^INK4a was also seen (r = 0.8.3, p < 0.001), there was also direct correlation between β2M and IL-1 β and IL-6 with (r = 0.5, p < 0.001; r = 0.68, p < 0.001) respectively. This evidence showed that β2M increased in buccal cells of the elderly compared to younger, and thereby buccal cells can be exploited to assess biological aging by measuring β2M levels, however, large sample size and using another assessing method such as β2M protein levels should be performed to confirm the results.