Affinity labeling of the androgen receptor with nonsteroidal chemoaffinity ligands

Mukherjee, Arnab; Kirkovsky, Leonid; Kimura, Yasuhiro; Marvel, Michele M.; Miller, Duane D.; Dalton, James T.

doi:10.1016/s0006-2952(99)00218-x

Cited by 23 publications

(44 citation statements)

References 23 publications

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“…Acetothiolutamide, R-bicalutamide, and R-iodo-bicalutamide were synthesized in our laboratories as described previously (Mukherjee et al, 1999;Kirkovsky et al, 2000). The purities of synthesized compounds were confirmed by elemental analysis and mass spectrometry.…”

Section: Methodsmentioning

confidence: 99%

“…Cytosolic AR was prepared from ventral prostates of castrated male Sprague-Dawley rats (about 250 g). The binding affinity of acetothiolutamide to the AR preparation was determined and analyzed as described previously (Mukherjee et al, 1996(Mukherjee et al, , 1999.…”

Section: Methodsmentioning

confidence: 99%

“…Nevertheless, the electrophilic character of the chloroacetamido group in R-1 raised a concern that it could potentially alkylate many nucleophilic sites in a cellular context. Although Scatchard analysis of AR binding demonstrated that R-1 is a reversible ligand for the AR (Mukherjee et al, 1999), it would be meaningful to further examine the possibility of non-electrophilic analogs of the lead antiandrogen pharmacophores as AR agonists. Therefore, we designed and synthesized acetothiolutamide (Fig.…”

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confidence: 99%

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Pharmacology, Pharmacokinetics, and Metabolism of Acetothiolutamide, a Novel Nonsteroidal Agonist for the Androgen Receptor

Yin¹,

Xu²,

He³

et al. 2002

J Pharmacol Exp Ther

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The present study characterized the in vitro androgen receptor (AR) binding affinity, in vitro and in vivo pharmacological activity, and in vivo pharmacokinetics and metabolism of acetothiolutamide, a nonsteroidal AR ligand. AR binding was determined by a competitive binding assay. In vitro AR agonist activity was examined by a cotransfection assay. Acetothiolutamide displayed high AR binding affinity (K i ϭ 4.9 Ϯ 0.2 nM) and full agonist activity in the in vitro studies. Next, the androgenic, anabolic, and antiandrogenic activity of acetothiolutamide was evaluated in a castrated immature rat model. In this animal model, acetothiolutamide exhibited an overall negligible androgenic effect, but a statistically significant anabolic effect at high subcutaneous doses. Also, acetothiolutamide demonstrated a noticeable antiandrogenic effect in castrated rats supplemented with testosterone propionate. To understand the causes for the observed disparity between in vitro and in vivo activities, pharmacokinetics and metabolism of acetothiolutamide were studied in male Sprague-Dawley rats. Acetothiolutamide was rapidly cleared from rat plasma (clearance of about 45 ml/min/kg) in a concentration-independent manner after i.v. dosing. Acetothiolutamide was completely absorbed after subcutaneous administration, but not bioavailable after oral dose. In the metabolism study, the unchanged molecule and its metabolites in urine and fecal samples were detected by highperformance liquid chromatography-mass spectrometry. The structures of major metabolites were elucidated with liquid chromatography-tandem mass spectrometry. After i.v. administration, acetothiolutamide was excreted in urine and feces as unchanged drug and a variety of metabolites. Oxidation, hydrolysis, and sulfate conjugation of phase I metabolites were the major metabolic pathways of acetothiolutamide in rats. Overall, the high plasma clearance of acetothiolutamide, due to its extensive hepatic metabolism, likely contributed to its lack of androgenic activity in vivo.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Pharmacology, Pharmacokinetics, and Metabolism of Acetothiolutamide, a Novel Nonsteroidal Agonist for the Androgen Receptor

Yin¹,

Xu²,

He³

et al. 2002

J Pharmacol Exp Ther

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“…During studies with affinity ligands for the AR, our group discovered a group of nonsteroidal androgens that are structural derivatives of bicalutamide and hydroxyflutamide, two known antiandrogens (Dalton et al, 1998;Mukherjee et al, 1999). Other laboratories have also reported the identification of nonsteroidal compounds that possess androgen activity (Dalton et al, 1998;Hamann et al, 1999;Negro-Vilar, 1999).…”

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confidence: 99%

Pharmacodynamics of Selective Androgen Receptor Modulators

Yin¹,

Gao²,

Kearbey³

et al. 2002

J Pharmacol Exp Ther

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The present study aimed to identify selective androgen receptor modulators (SARMs) with in vivo pharmacological activity. We examined the in vitro and in vivo pharmacological activity of four chiral, nonsteroidal SARMs synthesized in our laboratories. In the in vitro assays, these compounds demonstrated moderate to high androgen receptor (AR) binding affinity, with K i values ranging from 4 to 37 nM, and three of the compounds efficaciously stimulated AR-mediated reporter gene expression. The compounds were then administered subcutaneously to castrated rats to appraise their in vivo pharmacological activity. Androgenic activity was evaluated by the ability of these compounds to maintain the weights of prostate and seminal vesicle, whereas levator ani muscle weight was used as a measure of anabolic activity. The maximal response (E max ) and dose for half-maximal effect (ED 50 ) were determined for each compound and compared with that observed for testosterone propionate (TP). Compounds S-1 and S-4 demonstrated in vivo androgenic and anabolic activity, whereas compounds S-2 and S-3 did not. The activities of S-1 and S-4 were tissue-selective in that both compounds stimulated the anabolic organs more than the androgenic organs. These two compounds were less potent and efficacious than TP in androgenic activity, but their anabolic activity was similar to or greater than that of TP. Neither S-1 nor S-4 caused significant luteinizing hormone or follicle stimulating hormone suppression at doses near the ED 50 value. Thus, compounds S-1 and S-4 were identified as SARMs with potent and tissue-selective in vivo pharmacological activity, and represent the first members of a new class of SARMs with selective anabolic effects.

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“…1), which were analogous to bicalutamide 1a in structure. [1][2][3][4] Biological testing of these compounds exhibited high binding affinities and biological activities. 5-8 Based on the results obtained and homology modeling, we had earlier reported the binding interactions between the androgen receptor and the ligands.…”

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Synthesis of oxazolidinedione derived bicalutamide analogs

Nair

Mustafa

Mohler

et al. 2006

Tetrahedron Letters

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The synthesis of chiral oxazolidinedione derived bicalutamide analogs has been discussed. KeywordsOxazolidinedione; Bicalutamide; Enantiomers; Androgen; ReceptorIn our earlier investigations, we developed a series of second generation androgen receptor binding ligands 1b (Fig. 1), which were analogous to bicalutamide 1a in structure. 1-4 Biological testing of these compounds exhibited high binding affinities and biological activities. 5-8 Based on the results obtained and homology modeling, we had earlier reported the binding interactions between the androgen receptor and the ligands. 9-11 Examination of torsional strain parameters and hydrogen bonding interactions revealed that the molecule adopts a conformation similar to 1c. The hydroxyl group α to the carbonyl undergoes positioning in such a manner that it enters into hydrogen bonding interaction with Aspargine705 of the receptor. The amino group forms a weak hydrogen bond with the back bone oxygen of Leucine704. The computational predictions made were in agreement with the results obtained from the binding assays. However, to convincingly establish the aforesaid interactions and the conformation adopted by the molecules, it was highly imperative to have a suitable molecular model, which did not have free hydroxyl and amide protons and above all was conformationally restricted to a structure similar to 1c. Accordingly, we decided to interlock the amide nitrogen and the hydroxyl oxygen by a carbonyl functionality, which led to the design of structure 1d, an oxazolidinedione derivative. As of this day, no literature reports are available on the synthesis of chiral oxazolidinedione derived bicalutamide analogs and hence we devised and attempted a stereoselective synthesis of this molecule, the details of which form the subject matter of the present letter.Pyrrolidine-2-carboxylic acid 2 was acylated by Schotten Baumann reaction to obtain methacryloyl pyrrolidine-2-carboxylic acid 3, which was subsequently lactonized to obtain bromolactone 4, according to the procedure that we reported recently. 12,13 Nucleophilic substitution of compound 4 with 4′-fluorobenzenethiol under alkaline conditions in 2-propanol medium afforded compound 5. However, hydrolysis of compound 5 under acidic conditions did not afford the desired chiral hydroxy acid 6. Since our initial attempt failed to give 6, we decided to reinvestigate the synthesis of chiral hydroxy acid 6 (Scheme 1), as we knew from our earlier experiments that the hydrolysis of bromolactone 4 using HBr solution under reflux conditions would give 3-bromo-2-hydroxy-2-methylpropanoic acid 7 in excellent yields. 12Corresponding author. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptThe hydroxy and carboxyl groups of compound 7 were then protected with tribromoacetaldehyde under acidic conditions to obtain compound 8 ,14 which was coupled with 4′-fluorobenzenethiol. Acidification of the reaction mixture with concd HCl followed by workup and isolation afforded chiral hydroxy acid 6 in good...

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Affinity labeling of the androgen receptor with nonsteroidal chemoaffinity ligands

Cited by 23 publications

References 23 publications

Pharmacology, Pharmacokinetics, and Metabolism of Acetothiolutamide, a Novel Nonsteroidal Agonist for the Androgen Receptor

Pharmacology, Pharmacokinetics, and Metabolism of Acetothiolutamide, a Novel Nonsteroidal Agonist for the Androgen Receptor

Pharmacodynamics of Selective Androgen Receptor Modulators

Synthesis of oxazolidinedione derived bicalutamide analogs

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