Retinoic Acid US10188615, at-RA all-trans-retinoic acid at-RA BDBM323588 US10752616, Code No. atRA
- Liang, C; Qiao, G; Liu, Y; Tian, L; Hui, N; Li, J; Ma, Y; Li, H; Zhao, Q; Cao, W; Liu, H; Ren, X Overview of all-trans-retinoic acid (ATRA) and its analogues: Structures, activities, and mechanisms in acute promyelocytic leukaemia. Eur J Med Chem 220: (2021)
- ChEMBL_430393 (CHEMBL916377) Inhibition of ATRA-induced CYP26 in human MCF7 cells assessed ATRA as metabolism using [11.12-3H]-ATRA
- ChEMBL_430394 (CHEMBL916378) Inhibition of ATRA-induced CYP26 in human T47D cells assessed as ATRA metabolism using [11.12-3H]-ATRA
- ChEMBL_430395 (CHEMBL916379) Inhibition of ATRA-induced CYP26 in human T47D cell microsome assessed as ATRA metabolism using [11.12-3H]-ATRA
- ChEMBL_1462611 (CHEMBL3399274) Inhibition of CYP26A1 in ATRA-induced human HL60 cell microsomes incubated for 30 mins using ATRA and NADPH by HPLC method
- ChEBML_195330 Inhibition of [3H]ATRA binding to RAR alpha receptor
- ChEBML_196331 Inhibition of [3H]ATRA binding to RAR gamma receptor
- ChEMBL_195820 (CHEMBL800284) Inhibition of [3H]ATRA binding to RAR beta receptor
- ChEMBL_196331 (CHEMBL801568) Inhibition of [3H]ATRA binding to RAR gamma receptor
- ChEMBL_1517857 (CHEMBL3620653) Inhibition of CYP26A1 in ATRA-induced human HL60 cell microsomes incubated for 30 mins in dark condition with NADPH and ATRA by HPLC method
- ChEMBL_1793490 (CHEMBL4265409) Inhibition of chick RALDH2 expressed in doxycycline-inducible HEK293 cells assessed as inhibition of ATRA synthesis after 24 hrs in presence of ATRA and NAD+
- ChEBML_195317 Inhibition of [3H]ATRA-Hl60 binding to Retinoic acid receptor alpha
- ChEBML_195495 Inhibition of [3H]ATRA binding to retinoic acid receptor RAR beta
- ChEBML_195807 Inhibition of [3H]-ATRA-Hl60 binding to Retinoic acid receptor beta
- ChEBML_196318 Inhibition of [3H]ATRA-Hl60 binding to Retinoic acid receptor gamma
- ChEBML_196468 Binding affinity against retinoic acid receptor alpha by [3H]ATRA displacement.
- ChEBML_196588 Binding affinity against retinoic acid receptor beta by [3H]ATRA displacement.
- ChEBML_196589 Binding affinity against retinoic acid receptor gamma by [3H]ATRA displacement.
- ChEBML_197412 Inhibition of [3H]ATRA binding to retinoic acid receptor RAR alpha
- ChEBML_196005 Inhibition of [3H]ATRA binding to human Retinoic acid receptor RAR gamma
- ChEMBL_195306 (CHEMBL799862) Relative activity against Retinoic acid receptor RAR alpha compared to ATRA
- ChEMBL_195331 (CHEMBL877734) Inhibition of [3H]ATRA binding to Retinoic acid receptor RAR alpha
- ChEMBL_195490 (CHEMBL798912) Inhibition of [3H]-ATRA binding to baculovirus expressed RAR beta receptor
- ChEMBL_195494 (CHEMBL798915) Inhibition of [3H]ATRA binding to Retinoic acid receptor RAR beta
- ChEMBL_195495 (CHEMBL798916) Inhibition of [3H]ATRA binding to retinoic acid receptor RAR beta
- ChEMBL_195821 (CHEMBL800285) Inhibition of [3H]ATRA binding to Retinoic acid receptor RAR beta
- ChEMBL_196004 (CHEMBL797868) Inhibition of [3H]ATRA binding to baculovirus expressed RAR gamma receptor
- ChEMBL_196009 (CHEMBL798019) Inhibition of [3H]ATRA binding to Retinoic acid receptor RAR gamma
- ChEMBL_196010 (CHEMBL798020) Inhibition of [3H]-ATRA binding to Retinoic acid receptor RAR gamma
- ChEMBL_196332 (CHEMBL801569) Inhibition of [3H]ATRA binding to Retinoic acid receptor RAR gamma
- ChEMBL_197405 (CHEMBL799795) Inhibition of [3H]ATRA binding to baculovirus expressed RAR alpha receptor
- ChEMBL_197410 (CHEMBL799799) Inhibition of [3H]ATRA binding to Retinoic acid receptor RAR alpha
- ChEMBL_1793494 (CHEMBL4265413) Inhibition of RALDH2 in living intact choroid isolated from 4 day recovering chick eyes assessed as inhibition of ATRA synthesis after 24 hrs in presence of ATRA and NAD+
- ChEMBL_195184 (CHEMBL801817) Inhibition of [3H]ATRA binding to mouse Retinoic acid receptor RAR alpha
- ChEMBL_195491 (CHEMBL798913) Inhibition of [3H]-ATRA binding to human Retinoic acid receptor RAR beta
- ChEMBL_196005 (CHEMBL797869) Inhibition of [3H]ATRA binding to human Retinoic acid receptor RAR gamma
- ChEMBL_196471 (CHEMBL799618) Inhibition of [3H]ATRA binding to baculovirus expressed retinoid receptor RAR alpha
- ChEMBL_196659 (CHEMBL799694) Inhibition of [3H]ATRA binding to baculovirus expressed retinoid receptor RAR beta
- ChEMBL_196783 (CHEMBL799727) Inhibition of [3H]-ATRA binding to mouse Retinoic acid receptor RXR alpha
- ChEMBL_196928 (CHEMBL801228) Inhibition of [3H]ATRA binding to human Retinoic acid receptor RAR beta
- ChEMBL_197092 (CHEMBL806021) Inhibition of [3H]ATRA binding to human Retinoic acid receptor RAR alpha
- ChEMBL_197406 (CHEMBL799796) Inhibition of [3H]ATRA binding to human Retinoic acid receptor RAR alpha
- ChEMBL_197407 (CHEMBL799797) Inhibition of [3H]-ATRA binding to baculovirus expressed retinoid receptor RAR alpha
- ChEBML_195979 Ability to displace [3H]ATRA from Retinoic acid receptor gamma in CV-1 cells
- ChEBML_195982 Displacement of [3H]ATRA from RAR gamma receptor in CV-1 cells; Not tested
- ChEMBL_163472 (CHEMBL772307) Inhibition of [3H]ATRA binding to baculovirus expressed retinoic acid receptor RAR-alpha
- ChEMBL_163476 (CHEMBL772311) Inhibition of [3H]ATRA binding to baculovirus expressed retinoic acid receptor RAR-beta
- ChEMBL_163480 (CHEMBL772393) Inhibition of [3H]ATRA binding to baculovirus expressed retinoic acid receptor RAR-gamma
- ChEMBL_195328 (CHEMBL857604) Inhibition of [3H]ATRA binding to baculovirus expressed Retinoic acid receptor RAR alpha
- ChEMBL_195329 (CHEMBL802572) Inhibition of [3H]ATRA binding to baculovirus expressed Retinoic acid receptor RAR alpha
- ChEMBL_195470 (CHEMBL798147) Displacement of [3H]ATRA from RAR gamma receptor expressed in CV-1 cells
- ChEMBL_195817 (CHEMBL799666) Inhibition of [3H]ATRA binding to baculovirus expressed Retinoic acid receptor RAR beta
- ChEMBL_195818 (CHEMBL799667) Inhibition of [3H]ATRA binding to baculovirus expressed Retinoic acid receptor RAR beta
- ChEMBL_195819 (CHEMBL800283) Inhibition of [3H]ATRA binding to baculovirus expressed Retinoic acid receptor RAR beta
- ChEMBL_196328 (CHEMBL806271) Inhibition of [3H]ATRA binding to baculovirus expressed Retinoic acid receptor RAR gamma
- ChEMBL_196330 (CHEMBL801567) Inhibition of [3H]-ATRA binding to baculovirus expressed Retinoic acid receptor RAR gamma
- ChEMBL_52266 (CHEMBL665219) Inhibition of [3H]ATRA binding to chick skin Cytoplasmic retinoic acid binding protein
- ChEMBL_52394 (CHEMBL884347) Inhibition of [3H]ATRA binding to chick skin Cytoplasmic retinoic acid binding protein
- ChEBML_195468 Binding affinity against RAR beta receptor using [3H]ATRA as radioligand in CV-1 cells
- ChEBML_197251 Binding affinity against RAR alpha receptor using [3H]ATRA as radioligand in CV-1 cells
- ChEMBL_195982 (CHEMBL807688) Displacement of [3H]ATRA from RAR gamma receptor in CV-1 cells; Not tested
- ChEMBL_769532 (CHEMBL1833279) Inhibition of human CYP26A1 assessed using [11,12-3H]ATRA as substrate by scintillation counting
- ChEMBL_1736392 (CHEMBL4151928) Displacement of [3H]ATRA from 6H-AVI-GST-TEV-human RORbeta (T221 to K470 residues) pre-incubated for 30 mins before [3H]ATRA addition and measurted ater 18 to 23 hrs by scintillation proximity assay
- ChEBML_196011 Inhibition of 3[H]ATRA binding to retinoid acid receptor gamma expressed in CV-1 cells
- ChEBML_196324 Binding affinity towards retinoic acid receptor gamma was determined using [3H]ATRA (5 nM) as radioligand
- ChEMBL_195190 (CHEMBL802723) Binding affinity for Retinoic acid receptor alpha was determined by competing with 3[H]-ATRA
- ChEMBL_195644 (CHEMBL795962) Binding affinity for Retinoic acid receptor beta was determined by competing with 3[H]-ATRA
- ChEMBL_196168 (CHEMBL804941) Binding affinity for Retinoic acid receptor gamma was determined by competing with 3[H]-ATRA
- ChEMBL_52397 (CHEMBL665901) Inhibition of [3H]ATRA binding to murine Cytoplasmic retinoic acid binding protein (CRABP) type 1
- ChEMBL_52398 (CHEMBL665902) Inhibition of [3H]ATRA binding to murine Cytoplasmic retinoic acid binding protein (CRABP) type 1
- ChEMBL_52400 (CHEMBL665904) Inhibition of [3H]ATRA binding to murine Cytoplasmic retinoic acid binding protein (CRABP) type 2
- ChEMBL_52401 (CHEMBL875705) Inhibition of [3H]ATRA binding to murine Cytoplasmic retinoic acid binding protein (CRABP) type 2
- ChEMBL_1793492 (CHEMBL4265411) Inhibition of RALDH2 in choroidal cytosol fraction from 4 day recovering chick assessed as inhibition of ATRA synthesis pre-incubated for 20 mins followed by ATRA and NAD+ addition and measured after 30 mins by HPLC analysis
- ChEMBL_1517865 (CHEMBL3620661) Inhibition of CYP2D6 (unknown origin) incubated for 45 mins using NADPH and ATRA by HPLC assay
- ChEMBL_195324 (CHEMBL799719) Binding affinity towards retinoic acid receptor alpha was determined using [3H]ATRA (5 nM) as radioligand
- ChEMBL_195813 (CHEMBL807724) Binding affinity towards retinoic acid receptor beta was determined using [3H]ATRA (5 nM) as radioligand
- ChEMBL_196324 (CHEMBL806267) Binding affinity towards retinoic acid receptor gamma was determined using [3H]ATRA (5 nM) as radioligand
- ChEBML_195315 Antagonistic activity was evaluated in terms of inhibition of Retinoic acid receptor alpha transactivation by ATRA (50 nM)
- ChEMBL_196008 (CHEMBL797872) Binding affinity against Retinoic acid receptor gamma was determined in vitro by using [3H]ATRA as radioligand
- ChEMBL_196664 (CHEMBL803397) Tested for binding affinity against [3H]-ATRA binding to retinoid receptor isoform (RAR gamma) expressed in baculovirus
- ChEMBL_744044 (CHEMBL1768149) Inhibition of CYP26A1 in human MCF7 cell microsomes using [3H]ATRA after 1 hr by scintillation counting
- ChEBML_195806 Antagonistic activity was evaluated in terms of inhibition of Retinoic acid receptor beta transactivation by ATRA (50 nM); Not detectable
- ChEBML_196317 Antagonistic activity was evaluated in terms of inhibition of Retinoic acid receptor gamma transactivation by ATRA (50 nM); Not detectable
- ChEMBL_881496 (CHEMBL2214483) Inhibition of CYP26A1 in human MCF7 cell microsomes using [3H]ATRA as substrate after 1 hr by scintillation counter analysis
- ChEMBL_2502197 Antagonist activity at recombinant human RAR-beta expressed in mammalian cells in presence of ATRA by luciferase reporter gene based transactivation assay
- ChEMBL_2502198 Antagonist activity at recombinant human RAR-gamma expressed in mammalian cells in presence of ATRA by luciferase reporter gene based transactivation assay
- ChEMBL_848138 (CHEMBL2148657) Inhibition of CYP26A1-mediated retinoic acid metabolism in human MCF7 cell microsomes using [3H]ATRA as substrate after 1 hr by scintillation counting
- ChEMBL_2277619 Antagonist activity at Gal4-RARalpha transfected in HEK293 cells assessed as atRA-induced RARalpha transactivation and measured after 6 hrs by Dual-Light chemiluminiscence assay
- ChEMBL_1290847 (CHEMBL3118432) Antagonist activity at Gal4-fused RARalpha (unknown origin) transfected in HEK293 cells assessed as inhibition of ATRA-induced transcriptional activity by luciferase/beta-galactosidase reporter gene assay
- ChEMBL_2502186 Antagonist activity at human RAR-alpha transfected in human HeLa cells co-transfected with RARE-CAT reporter gene assessed as inhibition of ATRA-induced CAT expression measured after 24 hrs by transactivation competition assay
- ChEMBL_2502187 Antagonist activity at human RAR-beta transfected in human HeLa cells co-transfected with RARE-CAT reporter gene assessed as inhibition of ATRA-induced CAT expression measured after 24 hrs by transactivation competition assay
- ChEMBL_2502188 Antagonist activity at human RAR-gamma transfected in human HeLa cells co-transfected with RARE-CAT reporter gene assessed as inhibition of ATRA-induced CAT expression measured after 24 hrs by transactivation competition assay
- Transactivation assays Transcriptional transactivation assays were performed with gal4 fusion receptor constructs, created using each of the RAR ligand binding domains of either mouse or human, co-transfected with the pFR-luc (Stratagene) reporter construct in COS-7 cells. Thus, transfected cells will constitutively express the gal4-RAR fusion protein which in turn may be transactivated by all trans retinoic acid (atRA) to induce the expression of the luciferase that is driven by a gal4UAS.Briefly, on day 1, 96 well plates were seeded with 8000 cells per well then left to recover overnight. On day 2, the cells were co-transfected with 100 ng of reporter plasmid and 10 ng of the appropriate receptor plasmid per well using lipofectamine (Invitrogen). On day 3, the lipofectamine containing media was replaced by a DMEM without phenol red, followed by the addition of test compound dissolved in 1 μL of DMSO to each well's 100 μL total volume. Finally, on day 4, the cells were lysed and their luciferase substrate was provided by the BrightGlo reagent (Promega), the plates were then read on the MicroBeta TriLux (Perkin Elmer).On each plate, an 8 point dose-response curve of atRA was run in duplicate and dose-response curves of test compounds were also generated in duplicate.EC50 data both for test compounds and atRA was generated by fitting dose-response curves using GraphPad Prism . Data for test compounds are quoted as EC50 values. Where replicate data has been generated, the data are quoted as the mean EC50 from the separate experiments.
- In vitro CYP3A4 Inhibition Assay Cytochrome P450 is a large and diverse group of enzymes that catalyze the oxidation of organic substances. Some members of the CYP family contribute to the elimination of ATRA by catalyzing its 4-hydroxylation in the mammalian liver and skin, including that of humans as well as swine. Applicant evaluated the potential RAMBA activity of several azoles using pig liver microsomes, a rich source of CYP activity, comprising many different CYP 450 isoforms. Therefore, this approach, while a reasonable way to assess CYP inhibitors with broad activities may or may not be the best way to discover RAMBAs with selectivity for the skin, which has a much more narrow complement of CYP expression. As understanding in this area has progressed, a more specific CYP inhibition assay can be used to provide better predictivity of activity in human skin. Nevertheless, this assay may still be used as a general predictor of overall CYP activity.
- In vitro ATRA 4-Hydroxylase Activity Assay All procedures were carried out under minimal light in order to prevent degradation of the retinoid samples.Microsomal preparation: one lobe of fresh pig liver was obtained at the time of slaughter from a food-processing company and immediately placed in ice cold 15 mM KH2PO4/250 mM sucrose (pH 7.4) and kept on ice during transportation. A 10 g sample of liver was minced and homogenized in 30 mls of homogenization buffer (15 mM KH2PO4/250 mM sucrose) using a Tekmar homoginizer by pulsing 3 times 20 second pulses. This procedure was repeated for a total of 8x10 g samples of pig liver. The remaining pig liver was cut into 10-g pieces and wrapped in aluminum foil and stored at -80 °C. The homogenates from the 8 samples were pooled and centrifuged at 13,000xg for 20 minutes at 4 °C. to remove crude debris and the supernatant was further centrifuged at 100,000xg for 70 minutes at 4 °C. The microsomal pellets were resuspended into 50 mls of 150 mM KH2PO4/1 mM DTT (pH 7.4).