Synthesis and Biological Activities of 1α,4α,25- and 1α,4β,25-Trihydroxyvitamin D&lt;sub&gt;3&lt;/sub&gt; and Their Metabolism by Human CYP24A1 and UDP-Glucuronosyltransferase

Sawada, Daisuke; Tsukuda, Yuya; Yasuda, Kaori; Sakaki, Toshiyuki; Saito, Hiroshi; Takagi, Ken; Takenouchi, Kazuya; Chen, Thomas; Reddy, G. Satyanarayana; Kittaka, Atsushi

doi:10.1248/cpb.c12-00526

Cited by 11 publications

(10 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This is a totally different effect from that of the 4-OH group. 25) Although O2C3 was resistant to CYP24A1, its CYP3A4 metabolite 2 was further metabolized by CYP24A1, similar to 1. The k cat /K m value of hCYP24A1 for 2 was 60% of that for 1.…”

Section: Resultsmentioning

confidence: 99%

Synthesis and Biological Activity of 1α,2α,25-Trihydroxyvitamin D3: Active Metabolite of 2α-(3-Hydroxypropoxy)-1α,25-dihydroxyvitamin D3 by Human CYP3A4

et al. 2014

Self Cite

View full text Add to dashboard Cite

Our previous studies revealed that recombinant human CYP3A4 converted 2α-(3-hydroxypropoxy)-1α,25-dihydroxyvitamin D 3 (O2C3), which was a more potent binder to vitamin D receptor (VDR) than the natural hormone, 1α,25-dihydroxyvitamin D 3 (1α,25(OH) 2 D 3 , 1), to 1α,2α,25-trihydroxyvitamin D 3 (2). Here, we synthesized 2 using the Trost Pd-mediated coupling reaction between an A-ring precursor and a CD-ring bromoolefin and evaluated its preliminary biological activity. We found that metabolite 2 from O2C3 was still active as a VDR ligand while maintaining human VDR binding affinity (27 1-4) Actually, 1 and several synthetic analogs of 1 have been used clinically in the treatment of bone diseases, secondary hyperparathyroidism, psoriasis, and osteoporosis.1,5) Although 1 is inactivated by CYP24A1-dependent catabolism via C-24 hydroxylation to calcitroic acid for excretion from the body, 1) it was found that some 2α-substituted active vitamin D analogs were highly resistant to CYP24A1, for example, the k cat /K m value of 2α-(3-hydroxypropoxy)-1α,25-dihydroxyvitamin D 3 (O2C3), which showed 1.8-times greater binding affinity for vitamin D receptor (VDR) than 1, [6][7][8][9][10] was only 3% of that for 1. 11) CYP24A1 is the specific enzyme induced by the VDR-ligand (1 or its analog) complex in the target tissue and inactivates 1 and its analogs; therefore, CYP24A1-resistant ligands would have long-term biological effects on the target tissues.12) On the other hand, CYP3A4 is a broad-spectrum drug-metabolizing P450 enzyme, 13) but 1 and its analog 2α-(3-hydroxypropyl)-1α,25-dihydroxyvitamin D 3 (O1C3) are not primary substrates for CYP3A4.12) Recently, however, we demonstrated that O2C3 was metabolized by CYP3A4 and converted to 1α,2α,25-trihydroxyvitamin D 3 12) (2, Fig. 1). Its 2β-isomer (4) is a known compound and shows potent 1α,25(OH) 2 D 3 -like activities, 14) and we report here the synthesis of a new 2α-hydroxylated analog 2 using the Trost Pd-mediated coupling reaction between an A-ring precursor 12 and a CD-ring bromoolefin 13 to evaluate its preliminary biological activity. 15-18)The A-ring precursor 12 for Trost coupling was prepared from the known epoxide 5 19) in 11 steps (Chart 1). Briefly, treatment of 5 with p-methoxybenzyl alkoxide with heating gave methyl 3-O-(p-methoxybenzyl) altropyranoside 6, which had the 2α-hydorxy group (steroidal numbering) required in target molecule 2. After 2-O-silylation, the p-methoxybenzyl (PMB) protecting group was removed by 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) to give 7, and the methoxymethyl (MOM) group was introduced instead for the next bromination reaction using N-bromosuccinimide (NBS). NBS treatment for the resulting O-MOM-protected bezylidene acetal gave bromide 8. Activated Zn-reduction in the presence of NaBH 3 CN produced alcohol 9, which was converted to epoxide 10 via tosylation followed by tetrabutylammonium fluoride (TBAF) treatment. Trimethylsilyl (TMS)-ethynylation of 10 afforded enyne 11, and subsequent solvolysis in K 2 CO 3 -MeOH and O-...

show abstract

Section: Resultsmentioning

confidence: 99%

Synthesis and Biological Activity of 1α,2α,25-Trihydroxyvitamin D3: Active Metabolite of 2α-(3-Hydroxypropoxy)-1α,25-dihydroxyvitamin D3 by Human CYP3A4

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, the occurrence of the 4-hydroxylation of 1,25D 3 by CYP3A4 remains to be demonstrated and the biological significance of 4-hydroxymetabolites remains elusive. The only available biological data concern synthetic 1,4α,25D 3 and 1,4β,25D 3 ligands (Figure 1B) and the results of luciferase reporter assays in transfected human osteosarcoma cells indicate that both isomers are less active than 1,25D 3 [15,16]. Interestingly, significant differences in CYP24A1-induced 1,4,25D 3 metabolism have been observed between the two isomers, and only the 4α isomer has been shown to be glucuronidated by certain hepatic UGT(s) [16,17].…”

Section: Synthesis: General Informationmentioning

confidence: 99%

“…The only available biological data concern synthetic 1,4α,25D 3 and 1,4β,25D 3 ligands (Figure 1B) and the results of luciferase reporter assays in transfected human osteosarcoma cells indicate that both isomers are less active than 1,25D 3 [15,16]. Interestingly, significant differences in CYP24A1-induced 1,4,25D 3 metabolism have been observed between the two isomers, and only the 4α isomer has been shown to be glucuronidated by certain hepatic UGT(s) [16,17]. In addition, 4β,25D 3 has been shown to have greater metabolic stability and resistance to CYP24A1 than 4α,25D 3 [18].…”

Section: Synthesis: General Informationmentioning

confidence: 99%

4-Hydroxy-1α,25-Dihydroxyvitamin D3: Synthesis and Structure–Function Study

Peluso-Iltis,

Pierrat,

Rovito

et al. 2024

Biomolecules

Self Cite

View full text Add to dashboard Cite

The active vitamin D metabolites, 25-hydroxyvitamin D3 (25D3) and 1,25-dihydroxyvitamin D3 (1,25D3), are produced by successive hydroxylation steps and play key roles in several cellular processes. However, alternative metabolic pathways exist, and among them, the 4-hydroxylation of 25D3 is a major one. This study aims to investigate the structure–activity relationships of 4-hydroxy derivatives of 1,25D3. Structural analysis indicates that 1,4α,25(OH)3D3 and 1,4β,25(OH)3D3 maintain the anchoring hydrogen bonds of 1,25D3 and form additional interactions, stabilizing the active conformation of VDR. In addition, 1,4α,25D3 and 1,4β,25D3 are as potent as 1,25D3 in regulating the expression of VDR target genes in rat intestinal epithelial cells and in the mouse kidney. Moreover, these two 4-hydroxy derivatives promote hypercalcemia in mice at a dose similar to that of the parent compound.

show abstract

“…We have demonstrated that the E. coli expression system is quite useful to investigate enzymatic properties of CYP24A1. Using this E. coli expression system, we have determined kinetic parameters of CYP24A1 in the metabolism of the native vitamin D and various vitamin D derivatives, and revealed their metabolic pathways [45][46][47][48][49][50][51][52][53][54][55][56][57][58]. CYP24A1 plays central roles in vitamin D metabolism and produces a wide variety of metabolites.…”

Section: Construction Of a Cyp24a1 Enzyme System Containing Adrenodoxin (Adx) And Nadph-adrenodoxin Reductase (Adr)mentioning

confidence: 99%

Development of In Vitro and In Vivo Evaluation Systems for Vitamin D Derivatives and Their Application to Drug Discovery

Yasuda

Nishikawa

Mano

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

We have developed an in vitro system to easily examine the affinity for vitamin D receptor (VDR) and CYP24A1-mediated metabolism as two methods of assessing vitamin D derivatives. Vitamin D derivatives with high VDR affinity and resistance to CYP24A1-mediated metabolism could be good therapeutic agents. This system can effectively select vitamin D derivatives with these useful properties. We have also developed an in vivo system including a Cyp27b1-gene-deficient rat (a type I rickets model), a Vdr-gene-deficient rat (a type II rickets model), and a rat with a mutant Vdr (R270L) (another type II rickets model) using a genome editing method. For Cyp27b1-gene-deficient and Vdr mutant (R270L) rats, amelioration of rickets symptoms can be used as an index of the efficacy of vitamin D derivatives. Vdr-gene-deficient rats can be used to assess the activities of vitamin D derivatives specialized for actions not mediated by VDR. One of our original vitamin D derivatives, which displays high affinity VDR binding and resistance to CYP24A1-dependent metabolism, has shown good therapeutic effects in Vdr (R270L) rats, although further analysis is needed.

show abstract

Synthesis and Biological Activities of 1α,4α,25- and 1α,4β,25-Trihydroxyvitamin D<sub>3</sub> and Their Metabolism by Human CYP24A1 and UDP-Glucuronosyltransferase

Cited by 11 publications

References 8 publications

Synthesis and Biological Activity of 1α,2α,25-Trihydroxyvitamin D<sub>3</sub>: Active Metabolite of 2α-(3-Hydroxypropoxy)-1α,25-dihydroxyvitamin D<sub>3</sub> by Human CYP3A4

Synthesis and Biological Activity of 1α,2α,25-Trihydroxyvitamin D<sub>3</sub>: Active Metabolite of 2α-(3-Hydroxypropoxy)-1α,25-dihydroxyvitamin D<sub>3</sub> by Human CYP3A4

4-Hydroxy-1α,25-Dihydroxyvitamin D3: Synthesis and Structure–Function Study

Development of In Vitro and In Vivo Evaluation Systems for Vitamin D Derivatives and Their Application to Drug Discovery

Contact Info

Product

Resources

About