Victoria L. Challinor

Open-chain steroidal glycosides, a diverse class of plant saponins

Saponins are an important class of plant natural products that consist of a triterpenoid or steroidal skeleton that is glycosylated by varying numbers of sugar units attached at different positions. Steroidal saponins are usually divided into two broad structural classes, namely spirostanol and furostanol saponins. A third, previously unrecognized structural class of plant saponins, the open-chain steroidal saponins, is introduced in this review; these possess an acyclic sidechain in place of the heterocyclic ring/s present in spirostanols and furostanols. Open-chain steroidal saponins are numerous and structurally diverse, with over 150 unique representatives reported from terrestrial plants. Despite this, they have to date been largely overlooked in reviews of plant natural products. This review catalogs the structural diversity of open-chain steroidal saponins isolated from terrestrial plants and discusses aspects of their structure elucidation, biological activities, biosynthesis, and distribution in the plant kingdom. It is intended that this review will provide a point of reference for those working with open-chain steroidal saponins and result in their recognition and inclusion in future reviews of plant saponins.

Steroidal saponins from the roots of Smilax sp.: Structure and bioactivity

Phytochemical characterization of a commercial herb sample supplied as Smilax ornata Lem. (sarsaparilla) led to the isolation of five steroidal saponins, including two new furostanol saponins sarsaparilloside B (1) and sarsaparilloside C (2), whose structures were elucidated via a combination of multistage mass spectrometry (MS(n)), 1D and 2D NMR experiments, and chemical degradation. The previously unreported spectroscopic characterization of sarsaparilloside (3), Δ(20(22))-sarsaparilloside (4), and parillin (5) is also provided. The antiproliferative activity of the isolated saponins was compared in six human cell lines derived from different tumor types and one of the structures (2) was particularly active against the HT29 colon tumor cell line.

Carbon-Carbon Bond Cleavage in Activation of the Prodrug Nabumetone

Carbon-carbon bond cleavage reactions are catalyzed by, among others, lanosterol 14-demethylase (CYP51), cholesterol side-chain cleavage enzyme (CYP11), sterol 17β-lyase (CYP17), and aromatase (CYP19). Because of the high substrate specificities of these enzymes and the complex nature of their substrates, these reactions have been difficult to characterize. A CYP1A2-catalyzed carbon-carbon bond cleavage reaction is required for conversion of the prodrug nabumetone to its active form, 6-methoxy-2-naphthylacetic acid (6-MNA). Despite worldwide use of nabumetone as an anti-inflammatory agent, the mechanism of its carbon-carbon bond cleavage reaction remains obscure. With the help of authentic synthetic standards, we report here that the reaction involves 3-hydroxylation, carbon-carbon cleavage to the aldehyde, and oxidation of the aldehyde to the acid, all catalyzed by CYP1A2 or, less effectively, by other P450 enzymes. The data indicate that the carbon-carbon bond cleavage is mediated by the ferric peroxo anion rather than the ferryl species in the P450 catalytic cycle. CYP1A2 also catalyzes O-demethylation and alcohol to ketone transformations of nabumetone and its analogs.

NMR assignment of the absolute configuration of C-25 in furostanol steroidal saponins

The chemical shifts of the geminal proton resonances of H(2)-26 (δa and δb) are a widely used predictor of C-25 stereochemistry in furostanol steroidal saponins, being in general more resolved in 25S than 25R compounds. Unexpectedly, we found that application of this empirical rule in different solvents led to conflicting assignments of stereochemistry. An experimental survey revealed that, while the chemical shifts of H(2)-26 exhibit a dependence on C-25 configuration, it is less pronounced in methanol-d4 than pyridine-d5 solvent, and thus the general rule derived for pyridine-d5 fails when NMR spectra are acquired in methanol-d4. We propose a modified empirical method for the direct assignment of C-25 stereochemistry in furostanol saponins in methanol-d4 (Δ(ab)=0.45-0.48 ppm for 25S; Δ(ab)=0.33-0.35 ppm for 25R), and provide several detailed examples. In addition, the absolute configuration of compound 8, a steroidal saponin isolated in previous work from Ruscus colchicus, is corrected from 25R to 25S stereochemistry.

The Truth about False Unicorn (Chamaelirium luteum): Total Synthesis of 23R,24S-Chiograsterol B Defines the Structure and Stereochemistry of the Major Saponins from this Medicinal Herb

Chamaelirium luteum is used in traditional medicine systems and commercial botanical dietary supplements for the treatment of female reproductive health problems. Despite the wide use of this herb, only very limited phytochemical characterisation is available. Our investigation of C. luteum roots led to the isolation of two new steroidal saponins 1 and 2 that contain an unusual aglycone 3. The absolute configurations of these molecules were unable to be determined spectroscopically and thus the total synthesis of 3 was undertaken and achieved in 16 steps and 1.6 % overall yield from pregnenolone. The key step in the synthesis was the stereoselective installation of the side chain at C-17 and C-20, which employed anion-accelerated oxy-Cope methodology. The relative configuration of aglycone 3 was determined by X-ray crystallography of an advanced synthetic intermediate. The absolute configuration was based upon that of the pregnenolone-derived steroidal skeleton and determined to be 23R,24S.

Structure and Absolute Configuration of Helosides A and B, New Saponins from Chamaelirium luteum

Investigation of Chamaelirium luteum roots led to the isolation of two new steroidal saponins, helosides A and B, that contain a previously unreported aglycone, helogenin. Their structures and absolute configuration were elucidated through MS-MS, NMR, chemical degradation, and X-ray crystallography.

Revision of the absolute configurations of bethosides B and C and their aglycone.

The absolute stereochemistry of the steroidal saponins bethosides B and C was previously assigned as (22R,25R) on the basis of work that employed Horeaus method. Our studies of helosides A and B created doubt about both the original assignment and consequently our conclusion that relied upon it. The absolute configurations of bethosides B and C are revised to (22S,25R) following X-ray crystallographic analysis of their aglycone. Synthesis and full spectral characterization of both the 22R and 22S aglycones is reported to facilitate future stereochemical assignments in this series of saponins.

Structure and Absolute Configuration of Methyl (3R)-Malonyl-(13S)-hydroxycheilanth-17-en-19-oate, a Sesterterpene Derivative from the Roots of Aletris farinosa

We report the isolation and structure elucidation of a new cheilanthane sesterterpene from the roots of Aletris farinosa that possesses an unusual malonate half-ester functional group. The structure of 1 was determined via mass spectrometry and 1D and 2D NMR spectroscopy, while its absolute configuration was determined via X-ray crystallographic analysis performed on its methyl ester derivative 2.

Plant steroidal saponins: A focus on open-chain glycosides

Saponins are a large and structurally diverse class of phytochemicals that consist of a nonpolar steroidal or triterpenoid skeleton that is glycosylated by varying numbers of sugar residues at different positions. Steroidal saponins exhibit a large range of biological activities, including cytotoxic, anti-inflammatory, hemolytic, antifungal, and antibacterial properties. Saponins possessing a steroidal skeleton are usually divided into two main structural categories, namely, spirostanol and furostanol saponins. However, the open-chain steroidal glycosides represent a further structural class with numerous representatives, some of which possess potent cytotoxic activity. Different patterns of steroid oxygenation and glycosylation in furostanol, spirostanol, and open-chain steroidal saponins generate considerable structural diversity, which may account for their wide range of observed biological activities. The structures and stereochemistry of steroidal saponins are typically elucidated via a combination of multistage mass spectrometry, 1D and 2D NMR spectroscopy, and chemical degradation and synthesis.

Electrochemically mediated enantioselective reduction of chiral sulfoxides

The respiratory DMSO reductase from Rhodobacter capsulatus catalyzes the reduction of dimethyl sulfoxide to dimethyl sulfide. Herein, we have utilized this Mo enzyme as an enantioselective catalyst to generate optically pure sulfoxides (methyl p-tolyl sulfoxide, methyl phenyl sulfoxide and phenyl vinyl sulfoxide) from racemic starting materials. A hexaaminecobalt coordination compound in its divalent oxidation state was employed as the mediator of electron transfer between the working electrode and DMSO reductase to continually reactivate the enzyme after turnover. In all cases, chiral HPLC analysis of the reaction mixture revealed that the S-sulfoxide was reduced more rapidly leading to enrichment or isolation of the R isomer.Graphical Abstract