Production of 22-hydroxy metabolites of vitamin D 3 by cytochrome P450scc (CYP11A1) and analysis of their biological activities on skin cells

Robert C. Tuckey, Wei Li, Haleem Z. Shehabi, Zorica Janjetovic, Minh N. Nguyen, Tae Kang Kim, Jianjun Chen, Danielle E. Howell, Heather A.E. Benson, Trevor Sweatman, Donna M. Baldisseri, Andrzej Slominski

Research output: Contribution to journalArticle

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Abstract

Cytochrome P450scc (CYP11A1) can hydroxylate vitamin D 3, producing 20S- Mediated conversion to 20 Hydroxyvitamin D 3 [20(OH)D 3] and 20S,23- Di Hydroxyvitamin D 3 [20,23(OH) 2D 3] as the major metabolites. These are biologically active, acting as partial vitamin D receptor (VDR) agonists. Minor products include 17-hydroxyvitamin D 3, 17,20-dihydroxyvitamin D 3, and 17,20,23-trihydroxyvitamin D 3. In the current study, we have further analyzed the reaction products from cytochrome P450scc (P450scc) action on vitamin D 3 and have identified two 22-hydroxy derivatives as products, 22-hydroxyvitamin D 3 [22(OH)D 3] and 20S,22- dihydroxyvitamin D 3 [ 22Hydroxycholesterol, (20R,22R)20,22(OH) 2D 3]. The structures of both of these derivatives were determined by NMR. P450scc could convert purified 22(OH)D 3 to 20,22(OH) 2D 3. The 20,22(OH) 2D 3 could also be produced from 20(OH)D 3 and was metabolized to a trihydroxyvitamin D 3 product. We compared the biological activities of these new derivatives with those of 20(OH)D 3, 20,23(OH) 2D 3, and 1α,25- dihydroxyvitamin D 3 [1,25(OH) 2D 3]. 1,25(OH) 2D 3, 20(OH)D 3, 22(OH)D 3, 20,23(OH) 2D 3, and 20,22(OH) 2D 3 significantly inhibited keratinocyte proliferation in a dose-dependent manner. The strongest inducers of involucrin expression (a marker of keratinocyte differentiation) were 20,23(OH) 2D 3, 20,22(OH) 2D 3, 20(OH)D 3, and 1,25(OH) 2D 3, with 22(OH)D 3 having a heterogeneous effect. Little or no stimulation of CYP24 mRNA expression was observed for all the analogs tested except for 1,25(OH) 2D 3. All the compounds stimulated VDR translocation from the cytoplasm to the nucleus with 22(OH)D 3 and 20,22(OH) 2D 3 having less effect than 1,25(OH) 2D 3 and 20(OH)D 3. Thus, we have identified 22(OH)D 3 and 20,22(OH) 2D 3 as products of CYP11A1 action on vitamin D 3 and shown that, like 20(OH)D 3 and 20,23(OH) 2D 3, they are active on keratinocytes via the VDR, however, showing a degree of phenotypic heterogeneity in comparison with other P450scc-derived hydroxy metabolites of vitamin D 3.

Original languageEnglish (US)
Pages (from-to)1577-1588
Number of pages12
JournalDrug Metabolism and Disposition
Volume39
Issue number9
DOIs
StatePublished - Sep 1 2011

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Cholesterol Side-Chain Cleavage Enzyme
Hydroxycholecalciferols
Cholecalciferol
Calcitriol Receptors
Keratinocytes
Skin
Dihydroxycholecalciferols
Differentiation Antigens
Cytoplasm
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science

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Production of 22-hydroxy metabolites of vitamin D 3 by cytochrome P450scc (CYP11A1) and analysis of their biological activities on skin cells. / Tuckey, Robert C.; Li, Wei; Shehabi, Haleem Z.; Janjetovic, Zorica; Nguyen, Minh N.; Kim, Tae Kang; Chen, Jianjun; Howell, Danielle E.; Benson, Heather A.E.; Sweatman, Trevor; Baldisseri, Donna M.; Slominski, Andrzej.

In: Drug Metabolism and Disposition, Vol. 39, No. 9, 01.09.2011, p. 1577-1588.

Research output: Contribution to journalArticle

Tuckey, RC, Li, W, Shehabi, HZ, Janjetovic, Z, Nguyen, MN, Kim, TK, Chen, J, Howell, DE, Benson, HAE, Sweatman, T, Baldisseri, DM & Slominski, A 2011, 'Production of 22-hydroxy metabolites of vitamin D 3 by cytochrome P450scc (CYP11A1) and analysis of their biological activities on skin cells', Drug Metabolism and Disposition, vol. 39, no. 9, pp. 1577-1588. https://doi.org/10.1124/dmd.111.040071
Tuckey, Robert C. ; Li, Wei ; Shehabi, Haleem Z. ; Janjetovic, Zorica ; Nguyen, Minh N. ; Kim, Tae Kang ; Chen, Jianjun ; Howell, Danielle E. ; Benson, Heather A.E. ; Sweatman, Trevor ; Baldisseri, Donna M. ; Slominski, Andrzej. / Production of 22-hydroxy metabolites of vitamin D 3 by cytochrome P450scc (CYP11A1) and analysis of their biological activities on skin cells. In: Drug Metabolism and Disposition. 2011 ; Vol. 39, No. 9. pp. 1577-1588.
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abstract = "Cytochrome P450scc (CYP11A1) can hydroxylate vitamin D 3, producing 20S- Mediated conversion to 20 Hydroxyvitamin D 3 [20(OH)D 3] and 20S,23- Di Hydroxyvitamin D 3 [20,23(OH) 2D 3] as the major metabolites. These are biologically active, acting as partial vitamin D receptor (VDR) agonists. Minor products include 17-hydroxyvitamin D 3, 17,20-dihydroxyvitamin D 3, and 17,20,23-trihydroxyvitamin D 3. In the current study, we have further analyzed the reaction products from cytochrome P450scc (P450scc) action on vitamin D 3 and have identified two 22-hydroxy derivatives as products, 22-hydroxyvitamin D 3 [22(OH)D 3] and 20S,22- dihydroxyvitamin D 3 [ 22Hydroxycholesterol, (20R,22R)20,22(OH) 2D 3]. The structures of both of these derivatives were determined by NMR. P450scc could convert purified 22(OH)D 3 to 20,22(OH) 2D 3. The 20,22(OH) 2D 3 could also be produced from 20(OH)D 3 and was metabolized to a trihydroxyvitamin D 3 product. We compared the biological activities of these new derivatives with those of 20(OH)D 3, 20,23(OH) 2D 3, and 1α,25- dihydroxyvitamin D 3 [1,25(OH) 2D 3]. 1,25(OH) 2D 3, 20(OH)D 3, 22(OH)D 3, 20,23(OH) 2D 3, and 20,22(OH) 2D 3 significantly inhibited keratinocyte proliferation in a dose-dependent manner. The strongest inducers of involucrin expression (a marker of keratinocyte differentiation) were 20,23(OH) 2D 3, 20,22(OH) 2D 3, 20(OH)D 3, and 1,25(OH) 2D 3, with 22(OH)D 3 having a heterogeneous effect. Little or no stimulation of CYP24 mRNA expression was observed for all the analogs tested except for 1,25(OH) 2D 3. All the compounds stimulated VDR translocation from the cytoplasm to the nucleus with 22(OH)D 3 and 20,22(OH) 2D 3 having less effect than 1,25(OH) 2D 3 and 20(OH)D 3. Thus, we have identified 22(OH)D 3 and 20,22(OH) 2D 3 as products of CYP11A1 action on vitamin D 3 and shown that, like 20(OH)D 3 and 20,23(OH) 2D 3, they are active on keratinocytes via the VDR, however, showing a degree of phenotypic heterogeneity in comparison with other P450scc-derived hydroxy metabolites of vitamin D 3.",
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T1 - Production of 22-hydroxy metabolites of vitamin D 3 by cytochrome P450scc (CYP11A1) and analysis of their biological activities on skin cells

AU - Tuckey, Robert C.

AU - Li, Wei

AU - Shehabi, Haleem Z.

AU - Janjetovic, Zorica

AU - Nguyen, Minh N.

AU - Kim, Tae Kang

AU - Chen, Jianjun

AU - Howell, Danielle E.

AU - Benson, Heather A.E.

AU - Sweatman, Trevor

AU - Baldisseri, Donna M.

AU - Slominski, Andrzej

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N2 - Cytochrome P450scc (CYP11A1) can hydroxylate vitamin D 3, producing 20S- Mediated conversion to 20 Hydroxyvitamin D 3 [20(OH)D 3] and 20S,23- Di Hydroxyvitamin D 3 [20,23(OH) 2D 3] as the major metabolites. These are biologically active, acting as partial vitamin D receptor (VDR) agonists. Minor products include 17-hydroxyvitamin D 3, 17,20-dihydroxyvitamin D 3, and 17,20,23-trihydroxyvitamin D 3. In the current study, we have further analyzed the reaction products from cytochrome P450scc (P450scc) action on vitamin D 3 and have identified two 22-hydroxy derivatives as products, 22-hydroxyvitamin D 3 [22(OH)D 3] and 20S,22- dihydroxyvitamin D 3 [ 22Hydroxycholesterol, (20R,22R)20,22(OH) 2D 3]. The structures of both of these derivatives were determined by NMR. P450scc could convert purified 22(OH)D 3 to 20,22(OH) 2D 3. The 20,22(OH) 2D 3 could also be produced from 20(OH)D 3 and was metabolized to a trihydroxyvitamin D 3 product. We compared the biological activities of these new derivatives with those of 20(OH)D 3, 20,23(OH) 2D 3, and 1α,25- dihydroxyvitamin D 3 [1,25(OH) 2D 3]. 1,25(OH) 2D 3, 20(OH)D 3, 22(OH)D 3, 20,23(OH) 2D 3, and 20,22(OH) 2D 3 significantly inhibited keratinocyte proliferation in a dose-dependent manner. The strongest inducers of involucrin expression (a marker of keratinocyte differentiation) were 20,23(OH) 2D 3, 20,22(OH) 2D 3, 20(OH)D 3, and 1,25(OH) 2D 3, with 22(OH)D 3 having a heterogeneous effect. Little or no stimulation of CYP24 mRNA expression was observed for all the analogs tested except for 1,25(OH) 2D 3. All the compounds stimulated VDR translocation from the cytoplasm to the nucleus with 22(OH)D 3 and 20,22(OH) 2D 3 having less effect than 1,25(OH) 2D 3 and 20(OH)D 3. Thus, we have identified 22(OH)D 3 and 20,22(OH) 2D 3 as products of CYP11A1 action on vitamin D 3 and shown that, like 20(OH)D 3 and 20,23(OH) 2D 3, they are active on keratinocytes via the VDR, however, showing a degree of phenotypic heterogeneity in comparison with other P450scc-derived hydroxy metabolites of vitamin D 3.

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