Synthesis of α-santonin derivatives for diminutive effect on T and B-cell proliferation and their structure activity relationships.

A new library of 20 compounds from α-santonin was synthesized and tested against Con-A induced T-cell proliferation and LPS-induced B-cell proliferation via MTT assay. The study resulted in the identification of potent immunosuppressant molecules, which were further screened along with α-santonin for Tumor Necrosis Factor Alpha (TNF-α) inhibitory activity. One of the molecules (7) at 10 µM showed equipotency to that of dexamethasone (1 µM conc.) used as a standard. Structure activity relationships of the synthesized compounds along with our earlier reported α-santonin derivatives have been studied. Inferences from the modifications carried out at all the three sites of α-santonin have been elaborated. Computational study of the active compounds shows TNF-α protein as its preferable target rather than Inosine Monophosphate Dehydrogenase (IMPDH).

Biotransformation of tetrahydro-alpha-santonins by Absidia coerulea.

The fungus, Absidia coerulea was employed to bioconvert tetrahydro-alpha-santonins, 1,2,4alpha,5alpha-tetrahydro-alpha-santonin (1), and its 4-epimer (2), from which 10 products (3-12) were obtained. Furthermore, their structures were determined, based on their chemical and spectroscopic data analyses. Among them, 3-5, 7, 9, 11 and 12 were observed to be seven new compounds. The reactions mainly involved in these bio-process included hydroxylation(s) (C-4, C-11, and C-1), reduction (C-3 ketone to alcohol).

Biotransformation of alpha- and 6beta-santonin by fungus and plant cell cultures.

One fungus, Abisidia coerulea IFO 4011, and suspended cell cultures of one plant, Asparagus officinalis, were employed to bioconvert alpha- and 6beta-santonin. Incubation of alpha-santonin with the cell cultures of the fungus afforded two products, 11beta-hydroxy-alpha-santonin (1, in 76.5% yield) and 8alpha-hydroxy-alpha-santonin (2, in 2.0% yield). And from 6beta-santonin, four major products (3, 4, 5 and 6) and four minor products (7, 8, 9 and 10) were obtained, including 8alpha-hydroxylated products in trace yields. Very interestingly, a skeletal rearrangement occurred and a guaiane product (13) formed in a very low yield when alpha-santonin incubating with A.officinalis cell cultures, while not in the case of 6beta-santonin as substrate. Among the obtained 15 products, 2, 7, 8, 9, 10 and 12 are new compounds. The fact of 8alpha hydroxylation of santonin enables the formation of 8,12-eudesmanolide instead of 6,12-eudesmanolide and some useful modification at C-8 position. In addition, these reactions would provide evidence for the biogenesis between different types of eudesmane and/or guaiane compounds in the plants in nature.

Biotransformation of 6beta-santonin by Phytolacca acinosa cell suspension cultures.

AIM: To obtain more valuable derivatives for the further structural modification of 6beta-santonin (1) via biotransformation by using cell suspension cultures of Phytolacca acinosa. METHODS: The substrate 1 was incubated with cell suspension cultures of P. acinosa, the products were obtained by chromatography, and identified on the basis of their physical and spectral data (HRMS, 1D NMR, 2D NMR, NOE and IR). RESULTS: After incubation with cell suspension cultures of P. acinosa, 1 was converted into five products. Among them, 3 is a new compound. CONCLUSION: 6beta-santonin could be selectively reduced and hydroxylated by the cell suspension cultures of P. acinosa, which would provide valuable intermediates for its further structural modification.

Biotransformation of alpha-santonin by cell suspension cultures of five plants.

Cell suspension cultures of five plants (Catharanthus roseus, Ginkgo biloba, Platycodon grandiflorum, Taxus cuspidata, Phytolacca asinosa) were employed to bioconvert the eudesmanolide compound, alpha-santonin. Reactions occurring were hydroxylation (C-1, C-11 and C-15), reduction of the double bond [1(2) or 3(4)], rearrangment of the eudesmanolide skeleton to a guaianolide skeleton and lactone-ring hydrolysis. Four new compounds were identified.

Microbial transformations of alpha-santonin.

Fungal biotransformations of alpha-santonin (1) were conducted with Mucor plumbeus (ATCC 4740), Cunninghamella bainieri (ATCC 9244), Cunninghamella echinulata (ATCC 9245), Curvularia lunata (ATCC 12017) and Rhizopus stolonifer (ATCC 10404). Rhizopus stolonifer (ATCC 10404) metabolized compound 1 to afford 3,4-epoxy-alpha-santonin (2) and 4,5-dihydro-alpha-santonin (3) while Cunninghamella bainieri (ATCC 9244), Cunninghamella echinulata (ATCC 9245) and Mucor plumbeus (ATCC 4740) were capable of metabolizing compound 1 to give a reported metabolite, 1,2-dihydro-alpha-santonin (4). The structures of these transformed metabolites were established with the aid of extensive spectroscopic studies. These fungi regiospecifically reduced the carbon-carbon double bond in ring A of alpha-santonin.

Biotransformation of two cytotoxic terpenes, alpha-santonin and sclareol by Botrytis cinerea.

Two cytotoxic terpenes, alpha-santonin (1) and sclareol (3) were biotransformed by a plant pathogenic fungus Botrytis cinerea to produce oxidized metabolites in high yields. Alpha-Santonin (1) on fermentation with the fungus for ten days afforded a hydroxylated metabolite identified as 11beta-hydroxy-alpha-santonin (2) in a high yield (83%), while sclareol (3) was metabolized to epoxysclareol (4) (64%) and a new compound 8-deoxy-14,15-dihydro-15-chloro-14-hydroxy-8,9-dehydrosclareol (5) (7%), representing a rare example of microbial halogenation.

alpha-Santonin 1,2-reductase and its role in the formation of dihydrosantonin and lumisantonin by Pseudomonas cichorii S.

1,2-Dihydrosantonin is the first stable product in the degradative pathway of alpha-santonin by Pseudomonas cichorii S. Its formation is catalyzed by an oxidoreductase, which is NADH or NADPH dependent and has an apparent Km value of 66.66 microM for santonin and 44.33 microM for NADH. The enzyme activity is stable at pH 6.0, 7.0, and 8.0, and is not affected by EDTA and divalent metal ions. It is postulated that the enzymic reduction of santonin occurs via formation of a transient zwitterionic intermediate, which undergoes nonenzymatic 1,4-sigmatropic rearrangement to yield lumisantonin during the solvent extraction process. Lumisantonin is, thus, not a true metabolic intermediate but an artifact.

Studies on microbial transformations. XVIII. Microbiological transformation of 1-alpha-santonin.

The microbial transformation of a sesquiterpene lactone, 1-alpha-santonin was carried out by Streptomyces roseochromogenes NRRL-B-1233. The product obtained from 1-alpha-santonin was purified using silica gel column chromatography with 1% acetone-benzene as the eluant. The molecular weight of the product was 248, estimated by mass spectroanalysis. The product was assigned the chemical structure of 1,2-dihydro-1-alpha santonin on the bases of 1H and 13C-NMR and mass spectroanalyses. Similar products were also obtained from other actinomycetes. The conversion of 1-alpha-santonin to 1,2-dihydro-1-alpha-santonin was 28% for Streptomyces roseochromogenes NRRL-B-1233, 14% for S. roseochromogenes ATCC 13400, 30.9% for S. aureofaciens KCC-S-0008, and 43.0% for S. aureofaciens KCC-S-0624, respectively.

Enzyme-catalysed conjugations of glutathione with unsaturated compounds.

1. Rat-liver supernatant catalyses the reaction of diethyl maleate with glutathione. 2. Evidence is presented that the enzyme involved is different from the known glutathione-conjugating enzymes, glutathione S-alkyltransferase, S-aryltransferase and S-epoxidetransferase. 3. Rat-liver supernatant catalyses the reaction of a number of other alphabeta-unsaturated compounds, including aldehydes, ketones, lactones, nitriles and nitro compounds, with glutathione: separate enzymes may be responsible for these reactions.