Secosteroid biosynthesis

The C(23) epimers of 1alpha,23,25(OH)3-24-oxovitamin D3, a major natural metabolite of the secosteroid hormone, 1alpha,25(OH)2D3, were chemically synthesized for the first time. The metabolite was synthesized by palladium coupling of the appropriate CD ring analog with an A ring enyne. Various approaches from quinic acid to the A ring precursors were explored, and a new route to the A ring enyne from quinic acid was developed. The C(23) stereochemistry of the natural 1alpha,23,25(OH)3-24-oxovitamin D3 produced in neonatal human keratinocytes was determined to be S on the basis of the 1H NMR and the HPLC data. The biological activity of 1alpha,23(S), 25(OH)3-24-oxovitamin D3 in primary cultures of bovine parathyroid cells was determined by comparing the potency of this metabolite to that of 1alpha,25(OH)2D3 in suppression of parathyroid hormone (PTH) secretion. The results indicate that 1alpha,23(S), 25(OH)3-24-oxovitamin D3 potently suppressed PTH secretion even at concentrations as low as 10(-)12 M and is equipotent with 1alpha, 25(OH)2D3. The high activity of 1alpha,23(S),25(OH)3-24-oxovitamin D3 cannot be explained on the basis of its affinity for the vitamin D receptor as this metabolite was found to be 10 times less effective than radioinert 1alpha,25(OH)2D3 in blocking the uptake and receptor binding of [3H]-1alpha,25(OH)2D3 in intact parathyroid cells. Further studies are required to explain the molecular basis for the activity of 1alpha,23(S),25(OH)3-24-oxovitamin D3 in its ability to suppress PTH secretion. In summary, our present study indicates that the C(23) stereochemistry of the natural 1alpha,23, 25(OH)3-24-oxovitamin D3 is S and this metabolite is equipotent to 1alpha,25(OH)2D3 in suppressing PTH secretion.

Steroid isolation , depending on context, is the isolation of chemical matter required for chemical structure elucidation, derivitzation or degradation chemistry, biological testing, and other research needs (generally milligrams to grams, but often more [38] or the isolation of "analytical quantities" of the substance of interest (where the focus is on identifying and quantifying the substance (for example, in biological tissue or fluid). The amount isolated depends on the analytical method, but is generally less than one microgram. [39] [ page needed ] The methods of isolation to achieve the two scales of product are distinct, but include extraction , precipitation, adsorption , chromatography , and crystallization . In both cases, the isolated substance is purified to chemical homogeneity; combined separation and analytical methods, such as LC-MS , are chosen to be "orthogonal"—achieving their separations based on distinct modes of interaction between substance and isolating matrix—to detect a single species in the pure sample. Structure determination refers to the methods to determine the chemical structure of an isolated pure steroid, using an evolving array of chemical and physical methods which have included NMR and small-molecule crystallography . [2] : 10–19 Methods of analysis overlap both of the above areas, emphasizing analytical methods to determining if a steroid is present in a mixture and determining its quantity. [39]

1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) is a secosteroid hormone that renders dendritic cells (DCs) tolerogenic, favoring the induction of regulatory T cells. Induction of DCs with tolerogenic properties by 1,25(OH)(2)D(3) is associated with increased selective expression of immunoglobulin-like transcript 3 (ILT3), suggesting its involvement in the immunoregulatory properties of this hormone. Here we show an in vivo correlate of the increased ILT3 expression on DCs in healing psoriatic lesions following topical treatment with the 1,25(OH)(2)D(3) analog calcipotriol. Analysis of DC subsets reveals a differential regulation of ILT3 expression by 1,25(OH)(2)D(3), with a marked up-regulation in myeloid DCs but no effect on its expression by plasmacytoid DCs. A regulatory role for ILT3 expressed on DCs is indicated by the increased interferon-gamma (IFN-gamma) secretion promoted by anti-ILT3 addition to cultures of DCs and T cells, but this effect is blunted in 1,25(OH)(2)D(3)-treated DCs, suggesting ILT3-independent mechanisms able to regulate T-cell activation. Although ILT3 expression by DCs is required for induction of regulatory T cells, DC pretreatment with 1,25(OH)(2)D(3) leads to induction of CD4(+)Foxp3(+) cells with suppressive activity irrespective of the presence of neutralizing anti-ILT3 monoclonal antibody (mAb), indicating that ILT3 expression is dispensable for the capacity of 1,25(OH)(2)D(3)-treated DCs to induce regulatory T cells.

Secosteroid biosynthesis

secosteroid biosynthesis


secosteroid biosynthesissecosteroid biosynthesis