1232-73-1Relevant articles and documents
CYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate Modifications and Protein Engineering**
Bracco, Paula,Wijma, Hein J.,Nicolai, Bastian,Buitrago, Jhon Alexander Rodriguez,Klünemann, Thomas,Vila, Agustina,Schrepfer, Patrick,Blankenfeldt, Wulf,Janssen, Dick B.,Schallmey, Anett
, p. 1099 - 1110 (2021)
CYP154C5 from Nocardia farcinica is a P450 monooxygenase able to hydroxylate a range of steroids with high regio- and stereoselectivity at the 16α-position. Using protein engineering and substrate modifications based on the crystal structure of CYP154C5, an altered regioselectivity of the enzyme in steroid hydroxylation had been achieved. Thus, conversion of progesterone by mutant CYP154C5 F92A resulted in formation of the corresponding 21-hydroxylated product 11-deoxycorticosterone in addition to 16α-hydroxylation. Using MD simulation, this altered regioselectivity appeared to result from an alternative binding mode of the steroid in the active site of mutant F92A. MD simulation further suggested that the entrance of water to the active site caused higher uncoupling in this mutant. Moreover, exclusive 15α-hydroxylation was observed for wild-type CYP154C5 in the conversion of 5α-androstan-3-one, lacking an oxy-functional group at C17. Overall, our data give valuable insight into the structure–function relationship of this cytochrome P450 monooxygenase for steroid hydroxylation.
Identification and phenotype characterization of two CYP3A haplotypes causing different enzymatic capacity in fetal livers
Rodriguez-Antona, Cristina,Jande, Mary,Rane, Anders,Ingelman-Sundberg, Magnus
, p. 259 - 270 (2005)
Background: The fetal liver cytochrome P450 (CYP) 3A enzymes metabolize potentially toxic and teratogenic substrates and drugs in addition to endogenous hormones and differentiation factors. CYP3A7 is the most abundant CYP in the human liver during fetal stages and the first months of postnatal age and shows a large interindividual variability of unknown molecular basis. Methods: A new variant gene (CYP3A7* (*)2), which carries a mutation in exon 11 of CYP3A7 causing a T409R substitution, was identified by direct sequencing. Genotype analysis was performed by use of polymerase chain reaction followed by restriction enzyme analysis. CYP3A7.2 activity was assessed in heterologous expression systems and human fetal liver microsomes. Results: The frequency of CYP3A7* (*)2 was 8%, 17%, 28%, and 62% in white, Saudi Arabian, Chinese, and Tanzanian individuals, respectively. By use of human HEK293 cells, no significant differences in expression between CYP3A7.1 and CYP3A7.2 were found and fetal livers homozygous for CYP3A7* (*)2 had similar or higher CYP3A7 protein contents than CYP3A7* (*)1 livers. Kinetic studies showed that CYP3A7.2 was a functional enzyme with a significantly higher catalytic constant (kcat) as compared with CYP3A7.1 (P a linkage disequilibrium between the CYP3A7* (*)2 and CYP3A5* (*)1 alleles that was subject to interethnic differences. Determination of the alprazolam 1-hydroxylation rate revealed that CYP3A5 plays a significant role in the metabolism of CYP3A substrates in the fetal liver. Conclusion: We have identified 2 different CYP3A phenotypes in the fetal liver--one that is the result of a CYP3A7* (*)1/CYP3A5* (*)3 haplotype causing CYP3A7.1 but no CYP3A5 expression and another with higher detoxification capacity, inherent in the CYP3A7* (*)2/CYP3A5* (*)1 haplotype, where CYP3A5 and a more active form of CYP3A7 are expressed. Copyright
Novel components of the human metabolome: The identification, characterization and anti-inflammatory activity of two 5-androstene tetrols
Ahlem, Clarence N.,Page, Theodore M.,Auci, Dominick L.,Kennedy, Michael R.,Mangano, Katia,Nicoletti, Ferdinando,Ge, Yu,Huang, Yujin,White, Steven K.,Villegas, Sonia,Conrad, Douglas,Wang, Angela,Reading, Christopher L.,Frincke, James M.
, p. 145 - 155 (2011/03/18)
Two natural 5-androstene steroid tetrols, androst-5-ene-3β,7β, 16α,17β-tetrol (HE3177) and androst-5-ene-3α,7β,16α, 17β-tetrol (HE3413), were discovered in human plasma and urine. These compounds had significant aqueous solubility, did not bind or transactivate steroid-binding nuclear hormone receptors, and were not immunosuppressive in murine mixed-lymphocyte studies. Both compounds appear to be metabolic end products, as they were resistant to primary and secondary metabolism. Both were orally bioavailable, and were very well tolerated in a two-week dose-intensive toxicity study in mice. Anti-inflammatory properties were found with exogenous administration of these compounds in rodent disease models of multiple sclerosis, lung injury, chronic prostatitis, and colitis.
C19-Steroids as androgen receptor modulators: Design, discovery, and structure-activity relationship of new steroidal androgen receptor antagonists
Marwah, Padma,Marwah, Ashok,Lardy, Henry A.,Miyamoto, Hiroshi,Chang, Chawnshang
, p. 5933 - 5947 (2007/10/03)
Dehydroepiandrosterone (DHEA), the most abundant steroid in human circulating blood, is metabolized to sex hormones and other C19-steroids. Our previous collaborative study demonstrated that androst-5-ene-3β,17β-diol (Adiol) and androst-4-ene-3,17-dione (Adione), metabolites of DHEA, can activate androgen receptor (AR) target genes. Adiol is maintained at a high concentration in prostate cancer tissue; even after androgen deprivation therapy and its androgen activity is not inhibited by the antiandrogens currently used to treat prostate cancer patients. We have synthesized possible metabolites of DHEA and several synthetic analogues and evaluated their role in androgen receptor transactivation to identify AR modulators. Steroids with low androgenic potential in PC-3 cell lines were evaluated for anti-dihydrotestosterone (DHT) and anti-Adiol activity. We discovered three potent antiandrogens: 3β-acetoxyandrosta-1,5-diene-17-one 17-ethylene ketal (ADEK), androsta-1,4-diene-3,17-dione 17-ethylene ketal (OAK), and 3β-hydroxyandrosta-5,16-diene (HAD) that antagonized the effects of DHT as well as of Adiol on the growth of LNCaP cells and on the expression of prostate-specific antigen (PSA). In vivo tests of these compounds will reveal their potential as potent antiandrogens for the treatment of prostate cancer.
