Identification of benzoxazin-3-one derivatives as novel, potent, and selective nonsteroidal mineralocorticoid receptor antagonists.

Mineralocorticoid receptor (MR) blockade has come into focus as a promising approach for the treatment of cardiovascular diseases such as hypertension and congestive heart failure. In order to identify a novel class of nonsteroidal MR antagonists that exhibit significant potency and good selectivity over other steroidal hormone receptors, we designed a novel series of benzoxazin-3-one derivatives and synthesized them from 6-(7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazin-6-yl)-2H-1,4-benzoxazin-3(4H)-one (1a), high-throughput screening (HTS) hit compound. Our design was based on a crystal structure of an MR/compound complex and a docking model. In the course of lead generation from 1a, a 1,2-diaryl framework was characterized as a key structure with high binding affinity. On the basis of scaffold hopping and optimization studies, benzoxazin-3-one derivatives possessing 1-phenyl-3-trifluoromethylpyrazol-5-yl moiety at the 6-position were identified as a novel series of potent and selective MR antagonists. Among these compounds, 6-[1-(4-fluoro-2-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-5-yl]-2H-1,4-benzoxazin-3(4H)-one (14n) showed highly potent activity and good selectivity and also exhibited a significant antihypertensive effect in deoxycorticosterone acetate-salt hypertensive rats. On the basis of these results, compound 14n was progressed for further pharmacological evaluation.

[1]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[2]  G. Murshudov,et al.  Refinement of macromolecular structures by the maximum-likelihood method. , 1997, Acta crystallographica. Section D, Biological crystallography.

[3]  Jessica Huyet,et al.  Structural Basis of Spirolactone Recognition by the Mineralocorticoid Receptor , 2007, Molecular Pharmacology.

[4]  Kagawa Cm Blocking the renal electrolyte effects of mineralocorticoids with an orally active steroidal spirolactone. , 1960 .

[5]  Koichi Hayashi,et al.  Effectiveness of Aldosterone Blockade in Patients With Diabetic Nephropathy , 2003, Hypertension.

[6]  P. Raskin,et al.  Addition of angiotensin receptor blockade or mineralocorticoid antagonism to maximal angiotensin-converting enzyme inhibition in diabetic nephropathy. , 2009, Journal of the American Society of Nephrology : JASN.

[7]  N. Chapman,et al.  Effect of Spironolactone on Blood Pressure in Subjects With Resistant Hypertension , 2007, Hypertension.

[8]  K. Weber,et al.  Aldosterone in Congestive Heart Failure , 2004 .

[9]  Xu Shen,et al.  Synthesis, modification, and evaluation of (R)-de-O-methyllasiodiplodin and analogs as nonsteroidal antagonists of mineralocorticoid receptor. , 2011, Bioorganic & medicinal chemistry letters.

[10]  A. Vagin,et al.  MOLREP: an Automated Program for Molecular Replacement , 1997 .

[11]  C. Kagawa Blocking the renal electrolyte effects of mineralocorticoids with an orally active steroidal spirolactone. , 1960, Endocrinology.

[12]  W L Miller,et al.  Molecular biology of mineralocorticoid metabolism. , 1996, Annual review of nutrition.

[13]  K. Fujita,et al.  SM-368229, a novel selective and potent non-steroidal mineralocorticoid receptor antagonist with strong urinary Na+ excretion activity. , 2011, Journal of pharmacological sciences.

[14]  F. Kajiya,et al.  Involvement of Aldosterone and Mineralocorticoid Receptors in Rat Mesangial Cell Proliferation and Deformability , 2005, Hypertension.

[15]  Timothy M Willson,et al.  A Ligand-mediated Hydrogen Bond Network Required for the Activation of the Mineralocorticoid Receptor*[boxs] , 2005, Journal of Biological Chemistry.

[16]  Alexander Hillisch,et al.  A New Mode of Mineralocorticoid Receptor Antagonism by a Potent and Selective Nonsteroidal Molecule* , 2010, The Journal of Biological Chemistry.

[17]  Azra Mahmud,et al.  Does aldosterone-to-renin ratio predict the antihypertensive effect of the aldosterone antagonist spironolactone? , 2005, American journal of hypertension.

[18]  G. Stephenson,et al.  (S)-N-{3-[1-cyclopropyl-1-(2,4-difluoro-phenyl)-ethyl]-1H-indol-7-yl}-methanesulfonamide: a potent, nonsteroidal, functional antagonist of the mineralocorticoid receptor. , 2007, Journal of medicinal chemistry.

[19]  D. Cusi,et al.  Mechanisms of Disease: the role of aldosterone in kidney damage and clinical benefits of its blockade , 2007, Nature Clinical Practice Nephrology.

[20]  M. J. Meyers,et al.  Non-steroidal mineralocorticoid receptor antagonists , 2007 .

[21]  Z. Otwinowski,et al.  Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[22]  M. Young,et al.  Cardiac steroidogenesis in the normal and failing heart. , 2001, The Journal of clinical endocrinology and metabolism.

[23]  J. Ménard,et al.  Antialdosterones: incidence and prevention of sexual side effects. , 1989, Journal of steroid biochemistry.

[24]  D. Sica Pharmacokinetics and Pharmacodynamics of Mineralocorticoid Blocking Agents and their Effects on Potassium Homeostasis , 2005, Heart Failure Reviews.

[25]  B. Pitt,et al.  Eplerenone, a Selective Aldosterone Blocker, in Patients with Left Ventricular Dysfunction after Myocardial Infarction , 2003 .

[26]  B. Pitt,et al.  The effect of spironolactone on morbidity and mortality in patients with severe heart failure. Randomized Aldactone Evaluation Study Investigators. , 1999, The New England journal of medicine.

[27]  B. Pitt,et al.  The Effect of Spironolactone on Morbidity and Mortality in Patients with Severe Heart Failure , 2000 .

[28]  Henry Krum,et al.  A Comparison of the Aldosterone‐blocking Agents Eplerenone and Spironolactone , 2008, Clinical cardiology.

[29]  M. Heron,et al.  Discovery of (3S,3aR)-2-(3-chloro-4-cyanophenyl)-3-cyclopentyl-3,3a,4,5-tetrahydro-2H-benzo[g]indazole-7-carboxylic acid (PF-3882845), an orally efficacious mineralocorticoid receptor (MR) antagonist for hypertension and nephropathy. , 2010, Journal of medicinal chemistry.

[30]  J. Ménard,et al.  Efficacy and tolerance of spironolactone in essential hypertension. , 1987, The American journal of cardiology.

[31]  Claudine Mayer,et al.  Crystal structure of a mutant mineralocorticoid receptor responsible for hypertension , 2005, Nature Structural &Molecular Biology.

[32]  B. Roniker,et al.  Eplerenone: a selective aldosterone receptor antagonist (SARA). , 2006, Cardiovascular drug reviews.

[33]  P. Emsley,et al.  Features and development of Coot , 2010, Acta crystallographica. Section D, Biological crystallography.

[34]  Vincent B. Chen,et al.  Correspondence e-mail: , 2000 .