Multiple factors contribute to the toxicity of the aromatic retinoid, TTNPB (Ro 13-7410): binding affinities and disposition.
The aromatic retinoid (E)-4-[2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthylenyl)-1 -propenyl] benzoic acid (TTNPB) is 1000-fold more potent as a teratogen than all trans-retinoic acid (tRA) in several species and in the inhibition of chondrogenesis in the mouse limb bud cell culture. Factors responsible for the potency of TTNPB were investigated including binding to nuclear retinoic acid receptors (RARs and RXRs), cytosolic binding proteins (CRABPs), and metabolic disposition of TTNPB. For competitive binding assays and saturation kinetics, nucleosol or cytosol fractions were obtained from COS-1 cells transfected with cDNAs encoding the appropriate nuclear receptor or binding protein. TTNPB binds to RAR alpha, beta, and gamma with Kds in the nanomolar range; however, these binding affinities are 10-fold less than those of tRA. Although the affinities are high for TTNPB, it is unlikely that the binding affinities to nuclear receptors alone account for the potency of TTNPB. The binding affinities of TTNPB for the CRABPs are significantly lower than those of tRA. TTNPB did not compete with [3H]9-cis RA for binding to RXR alpha, beta, or gamma. Mouse limb bud cell cultures, a well characterized model for retinoid teratogenesis, were used to compare the metabolic disposition of TTNPB and tRA. In the media of limb bud cell cultures treated with either retinoid, the disappearance of TTNPB was significantly slower than that of tRA over 72 hr. Both retinoids reached approximately equal concentrations in cell uptake experiments; however, TTNPB disappeared from the limb bud cell at a significantly slower rate than did tRA. Collectively, these results indicate that high affinity binding to RARs, lower affinity to CRABPs, and resistance to metabolism contribute to the potency of TTNPB.
Pignatello
M A
MA
Laboratory of Cellular and Biochemical Toxicology, Rutgers University, Piscataway, New Jersey 08854, USA.
Kauffman
F C
FC
Levin
A A
AA
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D019540:Area Under Curve; D001565:Benzoates / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D001665:Binding Sites / Q000187:drug effects; D019556:COS Cells; D002356:Cartilage / Q000166:cytology / Q000187:drug effects; D046508:Culture Techniques; D003600:Cytosol / Q000378:metabolism; D005260:Female; D018878:Limb Buds / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D011247:Pregnancy; D018168:Receptors, Retinoic Acid / Q000378:metabolism*; D012176:Retinoids / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D014212:Tretinoin / Q000493:pharmacokinetics / Q000633:toxicity*
D000970:Antineoplastic Agents; D001565:Benzoates; D018168:Receptors, Retinoic Acid; D012176:Retinoids; D014212:Tretinoin; C038864:4-(2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-1-propenyl)benzoic acid