Lithocholic Acid Amides as Potent Vitamin D Receptor Agonists

Abstract

1α,25-Dihydroxyvitamin D3 [1α,25(OH)2D3, 1] is an active form of vitamin D3 and regulates various biological phenomena, including calcium and phosphate homeostasis, bone metabolism, and immune response via binding to and activation of vitamin D receptor (VDR). Lithocholic acid (LCA, 2) was identified as a second endogenous agonist of VDR, though its potency is very low. However, the lithocholic acid derivative 3 (Dcha-20) is a more potent agonist than 1α,25(OH)2D3, (1), and its carboxyl group has similar interactions to the 1,3-dihydroxyl groups of 1 with amino acid residues in the VDR ligand-binding pocket. Here, we designed and synthesized amide derivatives of 3 in order to clarify the role of the carboxyl group. The synthesized amide derivatives showed HL-60 cell differentiation-inducing activity with potency that depended upon the substituent on the amide nitrogen atom. Among them, the N-cyanoamide 6 is more active than either 1 or 3.

Vitamin D Receptor Mediates Attenuating Effect of Lithocholic Acid on Dextran Sulfate Sodium Induced Colitis in Mice

“…LCA derivatives, LCA acetate and LCA propionate, are VDR activators with less or no calcemic activity [ 50 ]. Thus, LCA derivatives may have potential for IBD therapy [ 51 ].…”

An N-Cyanoamide Derivative of Lithocholic Acid Co-Operates with Lysophosphatidic Acid to Promote Human Osteoblast (MG63) Differentiation

“…Next generation LCA amide derivatives look far more appealing for potential skeletal applications given their greater potency, matching, or even exceeding, their efficacy compared to 1,25D. We therefore turned our attention to a cyanoamide derivative of LCA, Dcha-150, given its compelling pro-differentiating effects on HL-60 cell cultures [11]. Herein, we report that MG63 osteoblasts co-stimulated with Dcha-150 and LPA/FHBP respond by exhibiting a clear and synergistic increase in ALP, a marker of the mature phenotype.…”

“…Since our original studies using LCA, the continued interest in this molec surrogate for 1,25D has stimulated the generation of other LCA derivatives w greater potency. These include the recent amide derivatives [11], especially th anoamide species, Dcha-150 (Figure 1), which has been found to elicit an HL-60 ferentiation response with an EC50 of 0.32 nm compared to 0.74 nm for 1,25D. A rable VDR transactivation was also observed for Dcha-150 in transfected human onic kidney (HEK293) cells.…”

“…Stocks of 1,25D (100 μM) were prepared in etha stored at −20 °C. Dcha-150 was synthesised as detailed previously [11], and stocks ( were prepared in DMSO and likewise stored at −20 °C. An inhibitor of the myo related transcription factor (MRTF)-serum response factor (SRF) gene transcriptio way, CCG 203971 (Bio-Techne Ltd., Abingdon, Oxford, UK), was prepared in eth mM) and also stored at −20 °C .…”

“…Unless stated otherwise, all reagents were of analytical grade from Sigma-Aldrich (Poole, UK). Stocks of 1-Oleoyl LPA (Bio-Techne Ltd., Abingdon, Oxford, UK) and FHBP (Tebu-Bio, Peterborough, UK), a phosphatase-resistant LPA analogue, were prepared in 1:1 ethanol:tissue culture grade water to a final concentration of 10 mM and 500 µM, respectively, and stored at −20 • C. Stocks of 1,25D (100 µM) were prepared in ethanol and stored at −20 • C. Dcha-150 was synthesised as detailed previously [11], and stocks (10 mM) were prepared in DMSO and likewise stored at −20 • C. An inhibitor of the myocardin-related transcription factor (MRTF)-serum response factor (SRF) gene transcription pathway, CCG 203971 (Bio-Techne Ltd., Abingdon, Oxford, UK), was prepared in ethanol (5 mM) and also stored at −20 • C. The methods identified below are essentially similar to those reported by us for related works [12][13][14][15].…”