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US007838552B2

(12) Patent (10) Patent No.: US 7,838,552 B2 Davis et al. (45) Date of Patent: Nov. 23, 2010

(54) COMPOSITIONS COMPRISING 2002.0068729 A1 6/2002 Egan et al. 2002/0107245 A1 8/2002 Wagle et al. (75) Inventors: Eric Davis, Morgantown, WV (US); 2002fO115655 A1 8/2002 Mehanna et al. John P. O'Donnell, Morgantown, WV 2002/O123485 A1 9, 2002 Alexander et al. (US); Peter Bruce Bottini, Morgantown, 2002fO147.184 A1 10/2002 Kosoglou et al. WV (US); Andrew Shaw, Morgantown, 2002fO151536 A1 10/2002 Davis et al. WV (US); R. Preston Mason, 2002fO161016 A1 10/2002 Tam et al. Manchester, MA (US) 2002fO169134 A1 11/2002 Davis 2002/0177586 A1 11/2002 Egan et al. (73) Assignee: Forest Laboratories Holdings Limited 2002/0183305 A1 12/2002 Davis et al. (BM) 2002/0183317 A1 12/2002 Wagle et al. 2002/0183365 A1 12/2002 Wagle et al. (*) Notice: Subject to any disclaimer, the term of this 2002/0192203 A1 12, 2002 Cho patent is extended or adjusted under 35 2003, OOO4194 A1 1, 2003 Gail U.S.C. 154(b) by 856 days. 2003, OO13699 A1 1/2003 Davis et al. 2003/0027820 A1 2, 2003 Gail (21) Appl. No.: 11/273,992 2003.0053981 A1 3/2003 Davis et al. (22) Filed: Nov. 15, 2005 2003, OO60489 A1 3/2003 Buckingham 2003, OO69221 A1 4/2003 Kosoglou et al. (65) Prior Publication Data 2003/0078517 A1 4/2003 Kensey US 2008/O161296A1 Jul. 3, 2008 2003/01 19428 A1 6/2003 Davis et al. 2003/01 19757 A1 6/2003 Davis Related U.S. Application Data 2003/01 19796 A1 6/2003 Strony 2003.01.19808 A1 6/2003 LeBeaut et al. (63) Continuation-in-part of application No. 1 1/141,235, 2003.01.19809 A1 6/2003 Davis filed on May 31, 2005. 2003/0158244 A1* 8, 2003 Devane et al...... 514,381 (60) Provisional application No. 60/577,423, filed on Jun. 2003,0162824 A1 8, 2003 Krul 4, 2004. 2003/0175344 A1 9, 2003 Waldet al. 2003/0176426 A1 9/2003 Wagle et al. (51) Int. Cl. A6 IK3I/352 (2006.01) CO7D 3II/04 (2006.01) (52) U.S. Cl...... 514/456; 514/1: 514/451 (Continued) (58) Field of Classification Search ...... 514/217.03, FOREIGN PATENT DOCUMENTS 5147456 See application file for complete search history. EP 1 400 529 3, 2004 (56) References Cited U.S. PATENT DOCUMENTS (Continued) 5,874,461 A 2/1999 De Chaffoy de Courcelles et al. OTHER PUBLICATIONS 5,922,341 A 7, 1999 Smith et al. 6,075,046 A 6/2000 De Chaffoy de Courcelles et al. Sato et al., “Current understanding of biochemical markers in heart 6,465,463 B1 10/2002 Cohn et al. failure.” MedSci Monit 2006; 12(11):RA252-246.* 6,495,154 B1 12/2002 Tam et al. 6,541479 B1 4/2003 Mehanna et al. (Continued) 6,545,040 B1 4/2003 Xhonneux et al. Primary Examiner Frederick Krass 6,595,926 B1 7/2003 Laragh Assistant Examiner Walter E Webb 6,596,744 B2 7/2003 Wagle et al. 6,596,745 B2 7, 2003 Gall (74) Attorney, Agent, or Firm—Charles Ryan; Michael 6,632,180 B1 10/2003 Laragh Ciraolo 6,635,273 B1 10/2003 Loscalzo et al. 6,770,663 B2 8/2004 Wagle et al. (57) ABSTRACT 6,784, 177 B2 8, 2004 Cohn et al. 6,869,966 B2 3/2005 Sato et al. 6,946,141 B2 9, 2005 Tam et al. Nebivolol has been shown to be beneficial in the treatment of 6,951,860 B2 10/2005 Mehanna et al. cardiovascular diseases such , congestive heart 7,030,106 B2 4, 2006 Cho failure, arterial stiffness and endothelial dysfunction. The 7,056,906 B2 6/2006 Strony present invention features a pharmaceutical composition 7,138.430 B2 11/2006 Garvey comprising nebivolol and at least one other active agent, 7,138,525 B2 11/2006 Sato et al. wherein the at least one other active agent is a cardiovascular 2001/0008896 A1 7/2001 Smith et al. agent. 2002fOO32149 A1 3/2002 Kensey 2002fOO44584 A1 4/2002 Fukunaga 2002fOO61835 A1 5/2002 Kensey 12 Claims, 4 Drawing Sheets US 7,838,552 B2 Page 2

U.S. PATENT DOCUMENTS WO WOO1/47509 T 2001 WO WOO1,76632 10, 2001 2003/0225146 Al 12/2003 Wagle et al. WO WOO1/97832 12/2001 2004.0005306 A1 1/2004 Loscalzo et al. WO WOO2,05851 1, 2002 2004/0023967 A1 2/2004 Cohn et al. WO WO O2/O7725 1, 2002 2004.0054177 A1 3, 2004 Otake et al. WO WO O2/O9760 2, 2002 2004, OO63646 A1 4/2004 Fujikura et al. WO WOO2.34303 5, 2002 2004f0071766 A1 4/2004 Loscalzo et al. WO WO O2/41883 5, 2002 2004f0071777 A1 4/2004 Trespidietal. WO WO O2/43806 6, 2002 2004/0097482 A1 5, 2004 Davis et al. WO WO O2/O53161 T 2002 2004/OO97495 A1 5/2004 Wagle et al. WO WO O2/O58685 8, 2002 2004/010.5850 A1 6/2004 Loscalzo et al. WO WO O2/O58731 8, 2002 2004.01098.94 A1 6, 2004 Shefer et al. WO WO O2/O58733 8, 2002 2004/O1328.05 A1 7/2004 Garvey WO WOO2O58731 8, 2002 2004/01763O8 A1 9, 2004 Shiohara et al. WO WO O2/O67851 9, 2002 2004/0176373 A1 9/2004 Buckingham WO WO O2/O58732 10, 2002 2004/0180860 A1 9, 2004 Burnett et al. WO WO O2/O875.08 11, 2002 2004/0180861 A1 9, 2004 Burnett et al. WO WO O2/O96362 12/2002 2004/O1974.82 A1 10, 2004 Lamaze et al. WO WO O2/O96363 12/2002 2004/O1987.00 A1 10, 2004 Burnett et al. WO WO O2/O964.15 12/2002 2004/0214811 A1 10, 2004 Davis et al. WO WOO3,O24456 3, 2003 2004/0235809 A1 11/2004 Alexander et al. WO WOO3,O26643 4/2003 2004/0235837 A1 1 1/2004 Egan et al. WO WO 03/026644 4/2003 2005, OO14747 A1 1/2005 Reinhard et al. WO WOO3,068.186 8, 2003 2005, OO31651 A1 2/2005 Gervais et al. WO WOO3,O926.17 11, 2003 2005/OO32788 A1 2/2005 Wagle et al. WO WOO3O926.17 11, 2003 2005/0032879 A1 2/2005 Okarter et al. WO WO 2004/O24720 3, 2004 2005/OO74487 A1 4/2005 Hsu et al. WO WO 2004/031175 4/2004 2005. O080071 A1 4/2005 Davis WO WO 2004/047837 6, 2004 2005, 0096307 A1 5, 2005 Graziano WO WO2004089416 10, 2004 2005/O153952 A1 T/2005 Cho WO WO2004.110375 12, 2004 2005/0215537 A1 9, 2005 Alexander et al. WO WO2005OOO217 1, 2005 2005/0222161 A1 10/2005 Moriya et al. WO WO 2005/046797 5, 2005 2005/0239766 A1 10, 2005 Starke et al. WO WO 2005/047285 5, 2005 2005/0272669 A1 12, 2005 Fushimi et al. WO WO 2005/065639 7/2005 2005/0272763 Al 12/2005 Toupence et al. WO WO 2005/099699 10/2005 2005/027281.0 A1 12, 2005 Davis et al. WO WO2005099699 10/2005 2006,0009399 A1 1/2006 Davis et al. WO WO 2005,102304 11, 2005 2006/0009513 A1 1/2006 Garvey WO WO 2005,107384 11, 2005 2006, OO 14828 A1 1/2006 Worcel et al. WO WO 2005,117591 12/2005 2006, OO 14829 A1 1/2006 Worcel et al. WO WO 2005,117858 12/2005 2006, OO24238 A1 2/2006 Barth et al. WO WO2005113012 12/2005 2006.0025352 A1 2/2006 Fujikura et al. WO WO 2006/O12642 2, 2006 2006, OO69080 A1 3, 2006 Veltri WO WO 2006/O15830 2, 2006 2006.0089349 A1 4/2006 Gundertofte et al. WO WO 2006/020244 2, 2006 2006/0094699 A1 5/2006 Kampen et al. WO WO 2006/O25070 3, 2006 2006/010O235 A1 5/2006 Andersen et al. WO WO2006060461 6, 2006 2006/0106008 A1 5/2006 Andersen et al. WO WO2006069293 6, 2006 2006/0106046 A1 5/2006 Moriya et al. WO WO2006084475 8, 2006 2006/01 10444 A1 5, 2006 Holmet al. WO WO2006084684 8, 2006 2006/011 1348 A1 5/2006 Kampen et al. WO WO2006086653 8, 2006 2006/011 1366 A1 5/2006 Andersen et al. WO WO2006 102069 9, 2006 2006/011 1380 A1 5, 2006 Otake et al. WO WO2006 102O71 9, 2006 2006, O142209 A1 6/2006 Nishimura et al. WO WO2006 102476 9, 2006 2006, O142288 A1 6, 2006 Peroutka WO WO2O06113485 11, 2006 2006, O148721 A1 7/2006 Erondu WO WO2006 115421 11, 2006

FOREIGN PATENT DOCUMENTS OTHER PUBLICATIONS EP 1405,859 4/2004 Maas et al., “Antihypertensive therapy: special focus on inter EP 1541 175 6, 2005 actions.” Expert Opin. Drug Saf. 2003:2(6):549-579.* EP 1544208 6, 2005 Ruzicka et al., “Monotherapy versus Combination therapy” EP 1548 024 6, 2005 2001:61(7);943-954.* EP 1550 668 7/2005 Cruickshank, J.M.. “Beta-blockers and : the bad guys come EP 1553 091 7/2005 good”. Cardiovascular Drugs and Therapy, vol. 16, No. 5, pp. 457 EP 1637 539 3, 2006 470, 2002. EP 1724OOO277 11, 2006 Whitworth, J.A., “Emerging drugs in the management of hyperten EP 1724.000278 11, 2006 sion”. Expert Opinion on Emerging Drugs, vol. 8, No. 2, pp. 377-388, WO WO 97.49394 12/1997 2003. WO WOOO.38653 T 2000 Kolck, et al., “Pharmacological basis of antihypertension drug WO WOO1? 12584 2, 2001 therapy”, vol. 93, No. 20, pp. 847-856, May 12, 2004. WO WOO1f17528 3, 2001 Cockcroft, J., “Nebivolol: a review.” Expert Opinion of Pharmacol WO WOO1/35961 5, 2001 ogy, vol. 5(4) (Jan. 1, 2004) pp. 893-899. US 7,838,552 B2 Page 3

European Search Report dated Jul. 28, 2008 for Application No. EP Drexler et al., “Correction of Endothelial Dysfunction in Coronary O8 OO 6021 Microcirculation of Hypercholesterolaemic Patients by L-”. Bruntun, T. Lauder. “Use of of Amyl in Pectoris', Lancet, 338: 1546-1550, 1991. London Hospital Medicine and Surgery, Jul. 1867,97-98. Nebivolol; Phase III Profiles Journal 2 (7); (Jul 1992) pp. 10-14. Vanhoutte, "Inhibition by of Creager, et al., "-Arginine Improves Endothelium-dependent neurotransmission in Vascular '. Circ. Res. in Hypercholesterolemic Humans'. American Society 1974:34:317-326. for Clinical Investigation, vol. 90, Oct. 1992, 1248-1253. Kamal, F., et al; "A pharmacokinetic and pharmocodynamic interac Malinski et al., “ release form a single measured in tion study between nebivolol and the H2- antagonists situ by a porphyrinic-based microsensor'. Letters to Nature, vol. 358, and ranitcline': 1977 Blackwell Science Ltd Br J. Clin Aug. 20, 1992. Pharmacol ; 43; pp. 201-204. Drexler et al., “Endothelial Function in Chronic Congestive Heart Gruetter, et al., “Relaxation of Bovine Coronary Artery and Activa Failure'. Am J Cardiol 69:1596-1601, 1992. tion of Coronary Arterial by Nitric Oxide, Taddei et al. “Endothelium-Dependent Forearm Vasodilation Is Nitroprusside and A Carcinogenic nitroSoamine'. Journal of Cyclic Reduced in Normotensive Subjects with Familial History of Hyper Research, 5(3):211-224, 1979. tension”, Journal of Cardiovascular , (Suppl 12):S193 Furchgott et al., “The obligatory role of endothelial cells in the S195, 1992. relaxation of arterial smooth muscle by acetylcholine”. Nature vol. Calver et al., “Effect of local intra-arterial N'-monomethyl-L- 288, Nov. 27, 1980. arginine in patients with hypertension: the nitric oxide dilator mecha Gryglewski, et al., Superoxide anion is involved in the breakdown of nism appears abnormal”, Journal of Hypertension, 10:1025-1031, endothelium-derived vascular relaxing factor, Letters to Nature, vol. 1992. 320, No. 3, Apr. 1986. Eighth Scientific Meeting of the American Society of Hypertension, Parthasarathy, et al., “ Inhibits Oxidative Modification of #1280 "Ambulatory Monitoring (ABPM) to Assess Low Density Lipoprotein'. The American Society for Clinical Inves the Hypertensive Effects of Nebivolol (N) and tigation, vol. 77, Feb. 1986. (HCTZ) Monotherapies and Combinations in Truly Hypertensive Harrison et al., “Alterations of Vascular Reactivity in Atherosclero Patients”, New York, New York, USA (May 19-22, 1993); American sis'. Circulation Research 1987; 61 (Suppl II):II-74-II-80. Journal of Hypertension 6 (5, part 2) p. 100A. Palmer, et al., "Nitric oxide release accounts for the biological activ Ford, G.A.; "Endothelial Dependent Venodilatory Action Of ity of endothelium-derived relaxing factor'. Letters to Nature, vol. Nebivolol In Human Hand Veins'; 6th European Meeting on Hyper 327, Jun. 1987. tension, Milan, , Jun. 4-7, 1993; pp. 239-240. Andrews, et al., “Low-density lipoproteins inhibit endothelium-de LeFabvre, J., et al., “Evaluation Du Nebivolol EtOu De pendent relaxation in rabbit aorta', vol. 327. May 1987. L'Hydrocholorothiazide vs Par Moniteur Ambulatoire: Bossaller, et al., “Impaired Muscarinic Endothelium-dependent Relaxation and Cyclic 5'-Monophosphate Formation in Devis Plurifactoriel'. Formule De Resume; (Oct. 20, 1993); 1 pg. Atherosclerotic Human Coronary Artery and Rabbit Aorta'. The Van Bortel, L. M. A. B.V., Beyond the tension of hypertension focus American Society for Clinical Investigation, vol. 79, 170-174, 1987. on non-antihypertensive of antihypertensive treatment Janssen Van Bortel and Rahn “Effect of nebivolol on the response of Research Foundation Publication (Nov. 18, 1993). in man' International Symposium on cardiovascular Taddei, et al., “Vasodilation to Acetylcholine in Primary and Second disease and B-adrenoceptor function Oct. 8 p. 54-65, 1988. ary Forms of Human Hypertension'. Hypertension 1993; 21:929 De Cree, et al. "An open pilot study on the effects of nebivolol in 5 933. patients with acute congestive ; International Sympo Casino, et al., “The Role of Nitric Oxide in Endothelium-Dependent sium on and B-adrenoceptor function, Oct. 8 Vasodilation of Hypercholerterolemic Patients'. Circulation, pp. 54-55, 1988. 1993;88:2541-2547. Pauwels, et al., “The Receptor Binding Profile of the New Ohara et al., “ Increases Endothelial Antihypertensive Agent Nebivolol and Its Stereoisomers Compared Superoxide Anion Production”. J. Clin. Invest. 91:2546-2551, 1993. with Various B-Adrenergis Blockers', Janssen Research Foundation, Lacourciere, et al., “Ambulatory blood pressure monitoring to assess Dept. of Biochemical Pharmacology, B-2340 Beerse, , Aug. the antihypertensive effects of nebivolol and hydrochlorotriazide 23, 1988. monotherapies and combinations in truly hypertensive patients'. Pauwels, et al., “B-Adrenoceptor-mediated cAMP accumulation in Amer. J. Hypertension 6: p. 100A, 1993. cardiac cells: effects of nebivolol”, European Journal of Pharmacol Lacourciere, and Arnott "Effects of nebivolol and low-dose ogy-Molecular Pharmacology Section 172 (1989) 417-479. hydrochlorothiazide on '. J. Hypertension 12 “Nebivolol; Drugs of the Future.” vol. 14; No. 10; 1989: pp.957-959. (Suppl. 3) p. 12, 1994. Panza, et al. "Abnormal Endothelium-dependent vascular relaxation Lacourciere, Yves, et al.; “Placebo-controlled comparison of the in patients with essential hypertension”, N Engl J Med., 1990: 323 effects of nebivolol and low-dose hydrochlorothiazide as 22-7. monotherapies and in combination on blood pressure and lipid profile Lu, HR., et al. “Effects of B-adrenoceptor antagoinists on cardiac in hypertensive patients'; Journal of Human Hypertension (1994) 8, function in Schemic-reperfused myocardium of the isolated working pp. 283-288. rabbit heart': European Journal of Pharmacology, 184 (1990) 65-74. Bowman, et al., "Nitric oxide mediated venodilator effects of Sieben, G., et al.; "Nebivolol in Hypertension . . . .” First Intern1 nebivolol'. British Journal of Clinical Pharmacology, 1994, 38:199 Nebivolol Investigators' Mtg. Antwerp, Belgium; Sep. 13-15, 1990; 204. Drug Investigation 3 (suppl. 1) pp. 193-195. Lacourciere, Yves, et al., “Treatment of Ambulatory Hypertensives Xhonneux et al., “The I-enantiomer of nebivolol potentiates the with Nebivolol or Hydrochlorothiazide Alone and in Combination”; blood pressure lowering effect of the d-enantiomer'. European Jour American Journal of Hypertension, Ltd. (Feb. 1994) 7:137-145. nal of Pharmacology 1990, 1181:261-265. 15th Scientific Meeting of the ISOH, Melbourne, (Mar. Gao, et al., “Nebivolol induces endothelium-dependent relaxations 20-24, 1994); Journal of Hypertension 12 (Suppl. 3) p. 12; "Effects of of canine coronary arteries'. Journal of Cardiovascular Pharmacol Nebivolol and Low Doses of Hydrochlorothiazide of LIPID Metabo ogy, 1991, 17:964-969. lism, A Placebo-Controlled Comparison”. Xi Journees de I’Hypertension Arterielle de la Societe Francaise de 15th Scientific Meeting of the ISOH, Melbourne, Australia (Mar. Cardiologie; Paris, , Dec. 12-13, 1991; Archives des Maladies 20-24, 1994); Journal of Hypertension 12 (Suppl. 3) p. 70. du coeur et des vaisseaux 84 p. 50. Cockcroft,et al., “Nebivolol vasodilates human forearm vasculature: Shimokawa, et al., Loss of Endothelial Pertussis Toxin-Sensitive G Evidence for an L-arginine/NO-dependent mechanism”, Journal of Function in Atherosclerotic Porcine Coronary Arteries, Cir Pharmacology and Experimental Therapeutics, 1995, 274: 1067 culation, 83:652-660, 1991. 1071. US 7,838,552 B2 Page 4

Treasure, et al., “Beneficial Effects of -Lowering Therapy Bouloumié et al., “Endothelial Dysfunction Coincides with an on the Coronary Endothelium in Patients with Coronary Artery Dis Enhanced Expression and Superoxide Anion ease'. The New England Journal of Medicine, vol. 332, No. 8, Feb. Production'. Hypertenion, 30:934-941, 1997. 23, 1995. McIntyre et al., “Sex Differences in the Abundance of Endothelial Anderson, et al. “The effect of cholesterol-lowering and antioxidant Nitric Oxide in a Model of Genetic Hypertension'. Hypertension, therapy on endotheoium-dependent coronary vasomotion'. The New 30:1517-1524, 1997. England Journal of Medicine, vol. 332, No. 8, Feb. 23, 1995. Mohri et al., “Basal Release of Nitric Oxide is Decreased in the Kamal, F., et al.; "The effects of and cimetidine on the Coronary Circulation in Patients with Heart Failure'. Hypertension, and pharmacodynamics of nebivolol; Proc. Brit. 30:50-56, 1997. Pharm. Soc., Oxford, UK, Jul. 12-14, 1995; pp. 519P-520P. Böger, et al., “Dietary -Arginine Reduces the Progression of Ath Liao, et al., “Oxidized Low-density Lipoprotein Decreases the erosclerosis in Cholesterol-Fed Rabbits', Circulation, 96:1282 Expression of Endothelial Nitric Oxide Synthase'. The Journal of 1290, 1997. Biological Chemistry, vol. 270, No. 1, pp. 319-324, Jan. 1995. Huk, et al., “-Arginine Treatment Alters the Kinetics of Nitric Oxide Burtet al., “Prevalance of Hypertension in the US Adult Population'. and Superoxide Release and Reduces Ischemia Reperfusion Injury in Hypertension, 25:305-313, 1995. ', Circulation, 96:667-675, 1997. Münzel, et al., “Evidence for Enhanced Vascular Superoxide Anion Zhang, et al., “ Releases Nitric Oxide From Canine Coro Production in Nitrate Tolerance'. J. Clin. Invest. 95:187-194, 1995. nary Microvessels”, Circulation, 97:576-580, 1998. Quyyumi, et al., “Contribution of Nitric Oxide to Metabolic Coro Vidal et al., “Atherogenic concentrations of native low-density nary Vasodilation in the Human Heart'. Circulation, 92:320-326, lipoproteins down-regulate nitric-oxide-synthase mRNA and protein 1995. levels in endothelial cells”, Eur J. Biochem, 252, 378-384 (1998). Mancini, et al., “-Converting Inhibition with Mason, et al., “Antioxidant and Cytoprotective Activities of the Cal Quinapril Improves Endothelial Vasomotor Dysfunction in Patients cium '. Biochemical Pharmacology, vol. with Coronary Artery Disease'. Circulation, 94:258-265, 1996. 55, pp. 1843-1852, 1998. Kugiyama, et al., “Nitric Oxide Activity Is Deficient in Spasm Arter Laufs, et al., “Upregulation of Endothelial Nitric oxide Synthase by ies of Patients with Coronary Spastic Angina'. Circulation, 94:266 HMG CA Reductase Inhibitors”, Circulation, 1998; 97: 1129-1135. 272, 1996. Kurtz et al., “Role of nitric oxide in the control of rennin secretion', Rajagopalan, et al., “Angiotensin II-mediated Hypertension in the American Physiological Society, 1998. Rat Increases Vascular Superoxide Production via Membrane Oemar, et al., “Reduced Endothelial Nitric Oxide Synthase Expres NADH/NADPH Oxidase Activation'. The Journal of Clinical Inves sion and Production in Human '. Circulation, tigation, Vo.97, No. 8, Apr. 1996. 97:2494-2498, 1998. Tulenko, et al., “Physical effects of cholesterol on arterial smooth Iadecola, et al., “Inducible Nitric Oxide Synthase Gene Expression in muscle membranes: evidence of immiscible cholesterol domains and Vascular Cells after Transient Focal Cerebral Ischemia', , alterations in bilayer width during atherogenesis'. Journal of Lipid 27:1373-1380, 1996. Research, vol.39, 1998. Münzel, et al., “Hydralazine Prevents Tolerance by Liao, 'Endothelium and acute coronary syndromes’. Clinical Chem Inhibiting Activation of a Membrane-bound NADH Oxidase'. The istry, 44:8(B) 1799-1808, 1998. Journal of Clinical Investigation, vol. 98, No. 6, 1465-1470, 1996. Monbouli et al., “Endothelial Dynsfunction: From Physiology to Shesely et al., “Elevated blood pressures in mice lacking endothelial Therapy”, J Moll Cell Cardiol 31:61-74, 1999. nitric oxide synthase'. Proc Natl Acad Sci USA, vol. 93, pp. 13176 Dawes, et al., “The vasodilator action of nebivolol in forearm 13181, Nov. 1996. vasculature of subjects with essential hypertension', British Journal Tschundi, et al., “Effect of Age on Kinetics of Nitric Oxide Release of Clinical Pharmacology, 1999, 48:460-463. in Rat Aorta and Pulmonary Artery'. The American Society for Cases, "Clinical Pharmacology of Nebivolol” Drugs of Today 1999, Clinical Investigation, vol. 98, No. 4, pp. 899-905, Aug. 1996. 35 (9); pp. 685-699. Mason, et al., “Effect of Oxidative Stress on Membrane Structure: McNeely, W. et al.; "Nebivolol in the Management of Essential Small-angle X-ray diffraction analysis”. Free Radical Biology & Hypertension\Review'. ADIS Drug Evaluation (Apr. 1999) 57 (4); Medicine, vol. 23, No. 3, pp. 419–425, 1997. pp. 633-651. Pieper, Galen M., "Acute Amelioration of Diabetic Endothelial Dys Freitag, A: "Nebivolol, a beta adrenoceptor antagonist with function with a Derivative of the Nitric oxide Synthase Cofactor, vasodilating effect': Pharmazeutische Zeitung Dec. 16, 1999, Ger ”, Journal of Cardiovascular Pharmacology, many, vol. 144, No. 50. 29:S-15, 1997. Janssen, et al., “Transient and Sustained impacts of hydroxyl radicals Harrison, David G. Perspective Series: Nitric Oxide and Nitric on sarcoplasmic reticulum function: protective effects of nebivolol”. Oxide Synthases, "Cellular and Molecular Mechanism of European Journal of Pharmacology, 1999, 366:223-232. Endothelial Cell Dysfunction'. American Society for Clinical Inves Kakoki, et al., “Effects of vasodilatory beta-adrenoceptor antagonists tigation, vol. 100, No. 9, Nov. 1997. on endothelium-derived nitric oxide release in rat '. Hyper Janssens, W.J., et al.; Symposium on: Endothelium in Hypertension, tension, Jan. 1999, 33(1, Pt. 2): 467-71. New Therapeutic Trends, “Pharmacology and pharmacokinetics of Brovkovych et al., “Nitric Oxide Release from Normal and Dysfunc nebivolol': Symposium on Endothelium in Hypertension, Berlin, tional Endothelium”, Journal of Physiology and Pharmacology, 50. (Mar. 1, 1997) pp. 10-13. 4, 575-586, 1999. Lvovich, et al., “Amperometric Sensors for Simultaneous Kellner-Weibel, et al., “Crystallization of Free Cholesterol in Model Superoxide and Hydrogen Peroxide Detection'. Analyticla Chemis Macrophage Foam Cells'. Arterioscler Thromb Vasc Biol. 19:1891 try, vol. 69, No. 3, Feb. 1997. 1898, 1999. Forte et al., “Basal nitric oxide synthesis in essential hypertension'. Broeders, et al., “Nebivolol: a third-generation B-blocker that aug Lancet 1997; 349:837-42. ments vascular nitric oxide release: endothelial beta(2)-adrenergis Stein, et al., “Vasodilation in black Americans: Attenuated nitric receptor-mediated nitric oxide production'. Circulation, 2000, oxide mediated responses'. Pharmacodynamics and Drug Action, 102:677-684. Clin Pharmacol Ther 62:436-43, 1997. Schachter, M.: “Nevivolol: how different, how interesting?"; British Pitei, et al., “NO-dependent Smooth Muscle Vasodilatation is Journal of Cardiology 7 (6), pp. 341, 343, 345, 347 (2000). Reduced in NIDDM Patients with Peripheral Sensory Neuropathy', Buga et al., Nitric Oxide: Biology and Chemistry Abstracts; vol. 4. Diabetic Medicine, 14:284-290, 1997. No. 3; p. 182 (2000); Academic Press, “Nebivolol: a possible novel Kinoshita, et al., “Inhibition of tetrahydrobiopterin mechanism in hypertension?: Drugs & Therapy Perspectives. impairs endothelium-dependent relaxations in canine basilar artery'. Drugs & Therapy Perspectives; "Nebivolol: Apossible novel mecha Tetrahydrrobiopterin Availabity and Endothelium, 1997. nism in hypertension'; vol. 16, No. 5 (2000). US 7,838,552 B2 Page 5

Erne, P: “Nebivolol': Pharma-Kritik 2000 , vol. 22, No. Sica, Dominic; Review Article, Drugs 2002; “Rationale for Fixed 8, 2000, pp. 31-32. Dose Combinations in the Treatment of Hypertension':vol. 62, (3), p. Parenti et al., “ phosphate metabolism and nitric-oxide 443-462. Synthase activity in endothelial cells are involved in the vasorelaxant Yakushin, S.S., et al.; “Assessment of the Safety and activity of nebivolol”, J Pharmacol Exp Ther, Feb. 2000; 292(2):698 Antihypertensive Effectiveness of the Cardioselective Beta TO3. Adrenoblocker Nebivolol in Patients with Arterial Hypertension Perregaux, et al., “Brachial Vascular Reactivity in Blacks'. Hyper Combined with Chronic Obstructive Bronchitis'; Kardiologia 2002, tension, 2000, 36:866-871. 11:36-39. Wagner, et al., “Improvement of Nitic Oxide-Dependent Vasodilata Zadionchenko, V.S., et al.; "Effects of Nevibolol on Microcircula tion by HMG-CoA Reductase Inhibitors Through Attenuation of tion, Platelet Aggregation and Blood Viscosity in Patients with Arte Endothelial Superoxide Anion Formation'. Arterioscler Thromb rial Hypertension'; Kardiologia 2002; 5:14-18. Vasc Biol. 20:61-69, 2000. Vertolli, U. et al.; “Eccentric LVH healing after starting renal replace John, et al., “Impaired endothelial function in arterial hypertension ment therapy” Journal of Nephrology 2002 Italy, vol. 15, No. 4, 2002, and hypercholesterolemia: potential mechanisms and differences'. pp. 403-405. Journal of Hypertension, 18:363-374, 2000. Kalinowski, et al., “Angiotensin IIAT Receptor Antagonists Inhibit Van Der Loo, et al., “Enhanced Peroxynitite Formation Is Associated Platelet Adhesion and Aggregation by Nitric Oxide Release'. Hyper with Bascular Aging”, J Exp Med, Vo. 192, No. 12, Dec. 18, 2000. tension, vol. 40: 521-527, 2002. Verganani et al., “Effect of Native and Oxidized Low-Density Kalinowski, et al., “ Potentiates Nitric Oxide Release and Lipoprotein on Endothelial Nitric Oxide and Superoxide Produc Enos Expression Through Inhibition of Isoprenoids Synthesis”,Jour tion’, Circulation, 101: 1261-1266, 2000. nal of Physiology and Pharmacology 2002: 53-4: 585-595. Troost, et al., “Nebivolol decreases systemic oxidative stress in Gourine, et al., " Antagonist Reduces healthy volunteers”. J. Clin Pharmacol. 50:377-379, 2000. Myocardial Reperfusion Injury by a Mechanism Related to Xue, et al., “Amperometric Ultramicrosensors for Peroxynitrite Bradykinin and Nitric Oxide'. Journal of Cardiovascular Pharmacol Detection and Its Application toward Single Myocardial Cells'. ogy, 40:564-570, 2002. Amer. Chem. Society, 72: 5313-5321, 2000. Landmesser, et al., “Role of p47' in Vascular Oxidative Stress and Gosgnach, W., et al.; "Nebivolol Induces Calcium-Independent Sig Hypertension Caused by Angiotensin II”, Hypertension, Oct. 2002. naling in Endothelial Cells by a Possible f-Adrenergic Pathway; Campia, et al., “Reduced Endothelium-Dependent and -Independent Journal of Cardiovascular Pharmacology, vol. 38, No. 2, 2001; pp. Dilation of Conductance Arteries in African Americans'. Journal of 191-199. American College of Cardiology, vol. 40, No. 4, 2002. 10th International Congress on Cardiovascular Pharmacotherapy, Cosentino, et al., "Nitric oxide-mediated relaxations in Salt-induced Kyoto, (Mar. 27-31, 2001); "Beneficial Effects of Combined hypertension: effect of chronic B-selective receptor blockade'. Jour Therapy Nebivolol vs. Only Trimetazidine in Heart nal of Hypertension, 20:421-428, 2002. Failure': Cardiovascular Drugs and Therapy 15 (Suppl. 1) p. 112. Pritchard, et al., “Native Low-Density Lipoprotein Induces Tzemos, Nikolaos, et al.; "Nebivolol Reverses Endothelial Dysfunc Endothelial Nitric Oxide Synthase Dysfunction: Role of Heat Shock tion in Essential Hypertension'; Circulation 104:511-5.14 (Jun. 8, Protein 90 and Caveolin-1”, Free Radical Biology & Medicine, vol. 2001). 33, No. 1, pp. 52-62, 2002. Asanuma et al., “, a Long-acting Ca Channel Blocker, Stepp, et al., “Native LDL and minimally oxidized LDL differentially Limits Infarct Size via Bradykinin- and NO-Dependent Mechanisms regulate Superoxide anion in vascular endothelium in situ'. Am J in Canine Hearts”. Cardiovascular Drugs & Therapy, 15 225-231. 2001. Physiol Heart Circ Physical, 283:H750-H759, 2002. Paniagua, et al., Role of Endothelial Nitric Oxide in Shear Stress Steinberg, et al., “Is the Oxidative Modification Hypothesis Relevant Induced Vasodilatino of Human Microvasculature, Circulation, to Human Atherosclerosis?', Circulation, 105:2107-2111, 2002. 103: 1752-1758, 2001. Ignarro, et al., "Nitric Oxide Donors and Cardiovascular Agents Loscalzo, Joseph, "Nitric oxide insufficiency, platelet activation, and Modulating the Bioactivity of Nitric Oxide: An Overview'. Circ. arterial thrombosis'. Circulation Research, 88:756-762, 2001. Res. 90:21-28, 2002. Martinez-Gonzales et al., “3-Hydroxy-3-Methylglutaryl Coenzyme Wassmann, et al., "Cellular Antioxidant Effects of In A Reductase Inhibition Prevents Endothelial NO Synthase Vitro and In Vivo”. Arterioscler Thromb Vasc Biol., 22:300-305, Downregulation by Atherogenic levels of Native LDLs”. Arterioscler 2002. Thromb Vasc Biol., 21:804-809, 2001. Bonetti et al., “Endothelial Dysfunction'. Arterioscler Thromb Vasc Napoli, et al., “Nitric Oxide and Atherosclerosis'. Nitric Oxide: Biol. , 23:168-175, 2003. Biology & Chemistry, vol. 5, No. 2, pp. 88-97, 2001. Bundkirchen et al., “B-adrenocetor selectivity of nebivolol and Feron et al., Hydroxy-Methylglutaryl-CoEnzyme A Reductase Inhi . A comparison of HICGP 12.177 and bition Promotes Endothelial Nitric Oxide Synthase Activation 'Iliodocyanopindolol binding studies”, European Journal of Phar Through a Decease in Caveolin Abundance, Circulation, 103: 113 macology 460 (2003) 19-26. 118, 2001. Jialal, et al., “Antioxidants and Atherosclerosis: Don't Throw Out the Falciani et al., “Effects of Nebivolol on Human Platelet Aggrega Baby with the Bath Water, Circulation 2003: 107:926-928. tion”, Journal of Cardiovascular Pharmacology, 38:922-929, 2001. Alekseeva, et al., “The use of nebivolol in menopausal women with Tanus-Santos, et al., “Effects of ethnicity on the distribution of clini hypertension', Kardiologiia, 2003, 43(10): 72-5. cally relevant endothelial nitric oxide variants'. Pharmacogenetics, Karimova, et al., “Efficiency of combined nebivolol and enalapril 719-725, 2001. therapy and their effect on regression of left hypertrophy'. Liu et al., “Effect of ACE Inhibitors and Angiotensin II Type 1 Uzbekiston Tibbiet Zhumali, (1), 2003, 46-48. Receptor Antagonists on Endothelial NO Synthase Knockout Mice Cominacini, et al., “Nebivolol and its 4-keto derivative increase nitric with Heart Failure'. Hypertension, 39 part 2:375-381, 2002. oxide in endothelial cells by reducing its oxidative inactivation'. Am Zou, et al., “Oxidation of the -thiolate complex and uncoupling Coll Cardiol. Nov. 19, 2003, 42(10): 1838-44. of endothelial nitric oxide synthase by peroxynitrite'. J. Clin Invest, Kalinowski, et al., “Third generation b-blockers stimulate nitric 109:817-826, 2002. oxide release from endothelial cells through ATP effux: a novel Gol Drin, et al., “Effect of enape combined with nebilet in hyperten mechanism for antihypertensive action'. Circulation, 2003, sion'. Lik Sprava, 2002, (7): 104-7. 107:2747-2752. Chlopicki, et al., “No-Dependent Vasodilation Induced by Nebivolol Mollnau, et al., “Nebivolol prevents vascular NOS III uncoupling in in Coronary Circulation is not Mediated by B-Adrenoceptors or by 5 experimental hyperlipidemia and inhibits NADPH oxidase activity in HT--Receptors”, Journal of Physiology and Pharmacology 2002, inflammatory cells'. Arteriosclerosis, Thrombosis, and Vascular 53, 4,615-624. Biology, 2003, 23:615-621. US 7,838,552 B2 Page 6

Izzo, Joseph et al.; IZZO and Black; Hypertension Primer, Third Mason, et al., “Effects of HMG-CoA Reductase Inhibitors on Edition, “The Essentials of High Blood Pressure”; 2003 p. 408-429; Endothelial Function'. Circulation, 109suppl. III:II-34-II-41, 2004. (Chapter C138). Tesloianu, et al., “Treatment of in COPD: is Amudha, K., et al.; “Ethnicity and Drug Therapy for Hypertension”; there hope for the better?', Pneumologia (Bucharest, Romania), Current Pharmaceutical Design, 2003, 9; pp. 1691-1701; 2003 53(4): 147-54, 2004. Bentham Science Publishers Ltd. Evdokimova, AG et al., “Nebivolol in the treatment of ischemic heart Rosei, E.A., et al.; “Evaluation of the and of disease patients with chronic heart failure'; Database accession No. Nebivolol versus—Lisinopril in the Treatment of Essential Arterial NLM15029131 abstract & Kardiologiia, 2004, vol. 44. No. 2, 2004, Hypertension: A Radomized, Multicentre'. Double-blind Study; pp. 15-18; US National Library of Medicine (NLM), Bethesda, MD. Blood Pressure 2003; 12 (Suppl. 1); 30-35. US; 2004. Rizos, E., et al.; "The Combination of nebivolol plus is De Groot, et al., “Antioxidant activity of nebivolol in the rat aorta', associated with a more beneficial metabolic profile compared to that Journal of Cardiovascular Pharmacology, 2004, 43:148-153. of plus pravastatin in hypertensive patients with Waring, W. S. et al.: “Is There a need for novel antihypertensive dyslipidemia': a pilot study; Journal of Cardiovascular Pharmacol therapies?”; Drug Discovery Today: Therapeutic Strategies, Elsevier, ogy and Therapeutics, Jun. 2003 v8 i2 p. 127(8); Westminster Pub lications Inc. vol. 1, No. 2, Oct. 2004, pp. 143-148. Lazar et al., “Beneficial effects of combined therapy nebivolol Cannon et al., “Intensive versus Moderate Lipid Lowering with trimetazidine, versus only trimetazidine in heart failure'. Cardiovas after Acute Coronary Syndromes”. The New England Journal cular Drugs and Therapy 15 (Supple. 1) p. 112, 2001. of Medicine, vol. 350, No. 15, Apr. 8, 2004. Gremmler, B. et al.; “Effects of AT1 therapy in Taylor, et al., “Combination of and Hydralazine patients with severe heart failure pretreated with angiotensin-con in Blacks with Heart Failure'. The New England Journal of Medi verting enzyme inhibitors'; Experimental and Clinical Cardiology cine, vol. 351, No. 20, Nov. 11, 2004. 2003 ; vol. 7, No. 4, 2003, pp. 193-198. Kalinowski, et al., “Race-Specific Differences in Endothelial Func Teoh, Y.P. et al.; "Candy in the clinic: Resistant hypertension in tion'. Circulation, 109:25.11-2517, 2004. diabetes' British Journal of Diabetes and Vascular Disease 2003, Boydak, et al.; "A Randomised Comparison of the Effects of , vol. 3, No. 4, 2003, pp. 300-301. Nebivolol and Atenolol with and without Chlorthalidone on the Katira and Chauhan "Nebivolol: A new on the horizon' Sexual Function of Hypertensive Men'; Clin. Drug Invest. 2005; Indian Heart J 52:86-88, (2000). 25(6) 409-416. Wing, et al. “Nebivolol and enalapril are not additive in combination Petersen, et al., “Trends in medicine”. American Society for Clinical in the treatment of essential hypertension' J. Hypertension 12 (Suppl. Pharmacology and Therapeutics, Mar. 3-5, 2005. 3) p. 70, (1994). Rosenkranz et al., “Phosphodiesterase type 5 inhibitor Van De Water, et al. “Pharmacological and Hemodynamic Profile of citrate does not potentiate the vasodilative properties of nebivolol in Nebivolol, a chemically novel, potent, and selective B1-adrenergic rat aorta'. Life Sciences 2005. antagonist” J. Cardiovascular Pharmacol 11: pp. 552-563, (1988). Mason et al., “Membrane location of nebivolol contributes to Thai, et al., “Angiotensin Subtype 1 Receptor (AT) Blockade antioxidant activity and endothelial nitric oxide release in stroke Improves Vasorelaxation in Heart Failure by Up-Regulation of prone hypertensive rats'. American Journal of Hypertension Endothelial Nitric-Oxide Synthase via Activation of the AT Recep 18:181A, 2005. tor'. Journal of Pharmacology and Experimental Therapeutics, Vop. 307, No. 3, 2003. Mason et al., “Nebivolol improves eNOS function and nitric oxide Landmesser, et al., “Oxidation of tetrahydrobioptgerin leads to in endothelial cells from African Americans'. Ameri uncoupling of endothelial cell nitric oxide synthase in hypertension'. can Journal of Hypertension, 18:181A, 2005. J. Clin Invest 111:1201-1209, 2003. Mason et al., “Nebivolol reduces nitroxidative stress and restores Mason, et al., “Membrane Microdomains and Vascular Biology”. nitric oxide bioavailability in endothelium of black Amerians'. Cir Circulation, 107:2270-2273, 2003. culation, 2005, 112:3795-3801. Ou, et al., “L-4F, an Apollipoprotein A-1 Mimetic, Restores Nitric Nissen, et al., Therapy, LDL Cholesterol, C-Reactive Protein, Oxide and Superoxide Anion Balance in Low-Density Lipoprotein and Coronary Artery Disease, The New England Journal of Medi Treated Endothelial Cells', Circulation, 107:1520-1524, 2003. cine, Jan. 6, 2005. Rodriguez-Porcel et al., “Hypercholesterolemia and Hypertension Gunnett, et al., “Mechanisms of Inducible Nitric Oxide Synthase Have Synergistic Deleterious Effects on Coronary Endothelial Func Mediated Vascular Dynfunction'. Arterioscler Tghromb Vasc Biol. tion', Arterioscler Thromb Vasc Biol., 23:885-891, 2003. 25:1617-1622, 2005. Wolfrum et al., “Endothelium-Dependent Effects of Statins”. Pasini, et al., “Nebivolol decreases oxidative stress in essential Arterioscler Thromb Vasc Biol., 23:729-736, 2003. hypertensive patients and increases nitric oxide by reducing its oxi Nissen et al., “Effect of Intensive Compared with Moderate Lipid dative inactivation”, J Hypertens 23:589-596, 2005. Lowering Therapy on Progression of Coronary Atherosclerosis'. Amer. Med. Assoc., vol. 291, No. 9, 2004. * cited by examiner U.S. Patent Nov. 23, 2010 Sheet 1 of 4 US 7,838,552 B2

Figure 1

560 Black + Nebivolol (1.0 uM) t t an White + Nebivolol (1.0 ulm) t D 500 Black, Untreated t atC 450 White, Untreated O O 400 350 C)s 300 O 250 O O 200 2 150 % o1 2N | | | | Untreated O 1.0 5.0 10 - Ramiprilat Treatment (uM) "p < 0.05 and tp < 0.01 vs preincubation with nebivolol alone (n = 6) U.S. Patent Nov. 23, 2010 Sheet 2 of 4 US 7,838,552 B2

Figure 2

250 Black Donors + Nebivolol (1.0 M) White Donors + Nebivolol (1.0M)

200

150

100

50

Ramiprilat Treatment (uM)

"p < 0.05 and tip < 0.01 vs preincubation with nebivolol alone (n = 6) U.S. Patent Nov. 23, 2010 Sheet 3 of 4 US 7,838,552 B2

Figure 3

500 Black + Nebivolol (1.0 uM) an OWhite + Nebivolol (1.0 uM) s 450 Black, Untreated e S White, Untreated .9C 400 350 as 300 O2 250 O 200 O Z 150 % o1 %N | | | | | | Untreated O 1.0 5.0 10 - Enalapril Treatment (uM) "p < 0.05 VS preincubation with nebivolol alone (n = 6) U.S. Patent Nov. 23, 2010 Sheet 4 of 4 US 7,838,552 B2

Figure 4

225 Black Donors + Nebivolol (1.0 uM) OWhite Donors + Nebivolol (1.0M)

3 200 C CD 175 CD (r. 150 O Z 125 S 100 CD 75 9 O 50 S 25 O O 10 5.0 10 Enalapril Treatment (uM)

"p < 0.05 vs preincubation with nebivolol alone (n = 6) US 7,838,552 B2 1. 2 COMPOSITIONS COMPRISING NEBVOLOL inhibitors, such as ), and mixtures thereof. In one embodiment, the other cardiovascular agent is an ACE inhibi This application is a continuation-in-part of application tor oran ARB. In a further embodiment, the other cardiovas Ser. No. 1 1/141,235, filed May 31, 2005, which is based on cular agent includes an ACE inhibitor and an ARB. In a and claims priority from U.S. Provisional Patent Application further embodiment, the ACE inhibitor is selected from the Ser. No. 60/577,423, Eric Davis, John O'Donnell, Peter Bot group consisting of alacepril, benazepril, , cero tini, filed Jun. 4, 2004. napril, cilaZapril, delapril, enalapril, enalaprilat, fosinopril, imidapril, lisinopril, perindopril, quinapril, ramipril, ramip TECHNICAL FIELD rilat, spirapril, temocapril, trandolapril. In a further embodi 10 ment, the ACE inhibitor is enalapril, ramipril, or ramiprilat.In This invention relates to compositions comprising nebiv a further embodiment, the other cardiovascular agent is an olol and one or more other active agent. More particularly, ARB Selected from the group consisting of candesartan, epro this invention relates to compositions comprising nebivolol Sartan, irbesartan, , Valsartan. and one or more cardiovascular agents for the treatment and/ In a further embodiment, the pharmaceutical composition or prevention of cardiovascular diseases. 15 comprises an amount of nebivolol in the range of between about 0.125 mg and about 40 mg. In a further embodiment, BACKGROUND OF THE INVENTION the amount of an ACE inhibitor may be in the range of Hypertension is a major health concern in the US. Approxi between about 0.5 mg to about 80 mg, and/or the amount of mately 50 million Americans have elevated blood pressure ARB may be in the range of between about 1 mg and about defined as a systolic blood pressure (SBP)2140 mmHg or a 1200 mg. diastolic blood pressure (DBP)290 mmHg. In addition, indi In a further embodiment, the pharmaceutical composition viduals with blood pressure of 120/80 mmHg or higher are at comprises nebivolol and only one other active agent. In a increased risk of developing hypertension and are considered further embodiment, the pharmaceutical composition com to be in a “pre-hypertension' state. Severity of hypertension is 25 prises nebivolol and only one cardiovascular agent. In a fur currently classified by stage, with Stage 1 hypertension span ther embodiment, the cardiovascular agent is selected from ning blood pressure ranges from 140/90 to 159/99 mmHg and the group consisting of ACE inhibitors (angiotensin II con Stage 2 including blood pressures: 160/100 mmHg. Verting enzyme inhibitors), ARB's (angiotensin II receptor antagonists), adrenergic blockers, adrenergicagonists, agents Onset of hypertension (diastolic alone or in combination for , anti-anginal agents, antiarrhyth with systolic) typically occurs between 25 and 55 years of 30 age. The risk of developing hypertension increases more dra mics, antiplatelet agents, , antihypertensives, matically with increasing age. According to the CDC, 68.3% antilipemic agents, antidiabetics, antiinflammatory agents, of men aged 65-74 have hypertension in the U.S. (Health blockers, CETP inhibitors, COX-2 inhibi United States, 2003, CDC/National Center for Health Statis tors, direct thrombininhibitors, diuretics, endothelin receptor tics) and 70.7% of menaged over 75 have hypertension in the 35 antagonists, HMG Co-A reductase inhibitors, inotropic U.S. (Health United States, 2003, CDC/National Center for agents, rennin inhibitors, vasodilators, vasopressors, AGE Health Statistics). In addition, 73.4% of women aged 65-74 crosslink breakers (advanced glycosylation end-product have hypertension in the US (Health United States, 2003, crosslink breakers, such as alagebrium, see U.S. Pat. No. CDC/National Center for Health Statistics) and 84.9% of 6,458.819), and AGE formation inhibitors (advanced glyco women aged over 75 have hypertension in the US (Health 40 Sylation end-product formation inhibitors, such as pimage United States, 2003, CDC/National Center for Health Statis dine). In a further embodiment, the active agent is an ACE tics). inhibitor or and ARB. Pharmaceutical formulations that stimulate, agonize, or In another aspect, the present invention features a method potentiate endothelial nitric oxide production, particularly of treating a subject for a cardiovascular disorder comprising formulations that produce increased nitric oxide levels in 45 administering to the subject an effective amount of nebivolol Black Americans, are needed. in combination with at least one other cardiovascular agent. In a further embodiment, the cardiovascular disorder is selected SUMMARY OF THE INVENTION from the group consisting of atherosclerosis, hypertension, diabetes mellitus, hyperhomocysteinemia, heart failure, and In one aspect, the present invention features a pharmaceu 50 renal failure. tical composition comprising nebivolol and at least one other In another aspect, the present invention features a method active agent. In a further embodiment, at least one of the of preventing a cardiovascular disorder comprising adminis active agents is a cardiovascular agent. In a further embodi tration to a subject an effective amount of nebivolol in com ment, the at least one cardiovascular agentis selected from the bination with an effective amount of at least one other car group consisting of ACE inhibitors (angiotensin II converting 55 diovascular agent. In a further embodiment, the enzyme inhibitors), ARB's (angiotensin II receptor antago cardiovascular disorder is selected from the group consisting nists), adrenergic blockers, adrenergic , agents for of congestive heart failure, hypertension, pulmonary hyper pheochromocytoma, antiarrhythmics, antiplatelet agents, tension, myocardial and cerebral infarctions, atherosclerosis, anticoagulants, antihypertensives, antilipemic agents, atherogenesis, thrombosis, ischemic heart disease, post-an antidiabetics, antiinflammatory agents, calcium channel 60 gioplasty restenosis, coronary artery diseases, renal failure, blockers, CETP inhibitors, COX-2 inhibitors, direct stable, unstable and variant (Prinzmetal) angina, cardiac inhibitors, diuretics, endothelin receptor antagonists, HMG , renal insufficiency, nephrotic edema, hepatic edema, Co-A reductase inhibitors, inotropic agents, rennin inhibi stroke, transient ischemic attacks, cerebrovascular accidents, tors, vasodilators, vasopressors, AGE crosslink breakers (ad restenosis, controlling blood pressure in hypertension, plate vanced glycosylation end-product crosslink breakers, such as 65 let adhesion, platelet aggregation, Smooth muscle cell prolif alagebrium, see U.S. Pat. No. 6,458.819), and AGE formation eration, , and vascular complications asso inhibitors (advanced glycosylation end-product formation ciated with the use of medical devices. US 7,838,552 B2 3 4 In another aspect, the present invention features a kitcom examples and appended claims are collected here. These defi prising an effective amount of nebivolol in combination with nitions should be read in light of the remainder of the disclo an effective amount of another cardiovascular agent. Sure and understood as by a person of skill in the art. Unless Even though nebivolol has 3-blocking properties, nebiv defined otherwise, all technical and scientific terms used olol is different from other classic f-blockers in that it is herein have the same meaning as commonly understood by a highly selective to the B1 adrenergic receptors and also has person of ordinary skill in the art. vasodilating effects related to its effect on endothelial nitric The articles “a” and “an are used herein to refer to one or oxide. It is believed that nebivolol increases the levels of nitric to more than one (i.e., to at least one) of the grammatical oxide within the vascular endothelium through the L-argin object of the article. By way of example, “an element’ means ine-nitric oxide pathway and has been shown to improve 10 one element or more than one element. endothelial dysfunction and improve compliance of blood About the same time” means that within about thirty min vessels. Nebivolol has also been shown to have antioxidant utes of administering one compound (nebivolol) to the characteristics which are favorable to the normal functioning patient, the other active compound(s) is/are administered to of the vascular endothelium. These characteristics make the patient. About the same time' also includes simultaneous nebivolol an effective antihypertensive agent with favorable 15 administration of the compounds. effects on the vascular endothelium and cardiovascular sys The phrase “angiotensin converting ” or tem. Nebivolol has been shown to be beneficial in the treat ACE inhibitor as used herein refers to a compound that ment of cardiovascular diseases such as hypertension, con inhibits any enzyme from converting angiotensin to any other gestive heart failure, arterial stiffness and endothelial form. dysfunction. In part, the present invention features a compo The phrase “angiotensin II receptor antagonist' or ARB' sition comprising nebivolol and at least one other cardiovas refers to a compound that binds to a receptor site on angio cular agent that is believed to work via a different mechanism tensin II but does not cause any physiological changes unless and is to be used for the treatment and/or prevention of vas another receptor is present. cular diseases characterized by nitric oxide insufficiency. The term “antagonist' is art-recognized and refers to a This invention also describes a method of reducing mortality 25 compound that binds to a receptor site, but does not cause a associated with cardiovascular disease in a blackpatient com physiological change unless another receptor ligand is prising administering to the black patient a therapeutically present. effective amount of nebivolol or its pharmaceutically salt and The term “bioavailable' is art-recognized and refers to a at least one other cardiovascular agent. This invention also form of the subject invention that allows for it, or a portion of relates to a method of improving NO release in a black patient 30 the amount administered, to be absorbed by, incorporated to, in need thereof by administering to the black patient a thera or otherwise physiologically available to a Subject or patient peutically safe and effective amount of nebivolol or a phar to whom it is administered. maceutically acceptable salt thereof and at least one other “Black” refers to a person of African descent or an African cardiovascular agent, sufficient to improve NO release. This American person but is not necessarily limited to those of invention further describes a method for improving exercise 35 African origin (e.g., Carribean). tolerance or for improving the quality of life in a blackpatient “Therapeutically effective amount” refers to the amount of in need thereof, comprising administering to the black patient the compound and/or composition that is effective to achieve a therapeutically effective amount of nebivolol or a pharma its intended purpose. ceutically salt thereof, and at least one cardiovascular agent. The phrase “cardiovascular agent” or “cardiovascular These embodiments of the present invention, other 40 drug” refers to a therapeutic compound that is useful for embodiments, and their features and characteristics, will be treating or preventing a cardiovascular disease. Non-limiting apparent from the description, drawings and claims that fol examples of Suitable cardiovascular agents include ACE low. inhibitors (angiotensin II converting enzyme inhibitors), ARB's (angiotensin II receptor antagonists), adrenergic BRIEF DESCRIPTION OF THE DRAWINGS 45 blockers, adrenergic agonists, agents for pheochromocy toma, agents, antiarrhythmics, antiplatelet FIG. 1 depicts a comparison of NO release from Black and agents, anticoagulants, antihypertensives, antilipemic agents, White donor endothelial cells after chronic treatment with antidiabetics, antiinflammatory agents, calcium channel ramprilat followed by treatment with nebivolol (1 uM). blockers, CETP inhibitors, COX-2 inhibitors, direct thrombin FIG. 2 depicts a comparison of the increase in NO release 50 inhibitors, diuretics, endothelin receptor antagonists, HMG from Black and White donor endothelial cells after chronic Co-A reductase inhibitors, inotropic agents, inhibitors, treatment with ramiprilat followed by treatment with nebiv vasodilators, vasopressors, AGE crosslink breakers (ad olol (1 uM). vanced glycosylation end-product crosslink breakers, such as FIG. 3 depicts comparison of NO release from Black and alagebrium, see U.S. Pat. No. 6,458.819), and AGE formation White donor endothelial cells after chronic treatment with 55 inhibitors (advanced glycosylation end-product formation enalapril followed by treatment with nebivolol (1 uM). inhibitors, such as pimagedine), and combinations thereof. FIG. 4 depicts a comparison of the increase in NO release Cardiovascular disease or disorder refers to any cardiovas from Black and White donor endothelial cells after chronic cular disease or disorder known in the art, including, but not treatment with enalapril followed by treatment with nebivolol limited to, wherein the cardiovascular disease is selected (1 uM). 60 from the group consisting of congestive heart failure, hyper tension, pulmonary hypertension, myocardial and cerebral DETAILED DESCRIPTION OF THE INVENTION infarctions, atherosclerosis, atherogenesis, thrombosis, ischemic heart disease, post-angioplasty restenosis, coronary Definitions artery diseases, renal failure, stable, unstable and variant 65 (Prinzmetal) angina, cardiac edema, renal insufficiency, For convenience, before further description of the present nephrotic edema, hepatic edema, stroke, transient ischemic invention, certain terms employed in the specification, attacks, cerebrovascular accidents, restenosis, controlling US 7,838,552 B2 5 6 blood pressure in hypertension, platelet adhesion, platelet organ, orportion of the body. Each carrier must be acceptable aggregation, Smooth muscle cell proliferation, pulmonary in the sense of being compatible with the Subject composition edema, and vascular complications associated with the use of and its components and not injurious to the patient. Some medical devices. examples of materials which may serve as pharmaceutically The term “combination” refers to two or more different acceptable excipients include: (1) Sugars, such as lactose, active agents which are administered at roughly about the glucose and Sucrose; (2) starches, such as corn starch and same time (for example, where the active agents are in a single potato starch; (3) cellulose, and its derivatives, such as pharmaceutical preparation) or at different times (for sodium carboxymethyl cellulose, ethyl cellulose and cellu example, one agent is administered to the Subject before the lose ; (4) powdered tragacanth; (5) malt, (6) gelatin: other). 10 (7) talc.; (8) excipients, such as cocoa butter and Suppository The terms “drug.” “pharmaceutically active agent.” “bio waxes; (9) oils, such as peanut oil, cottonseed oil, safflower active agent.” “therapeutic agent, and “active agent may be oil, sesame oil, olive oil, corn oil and Soybean oil: (10) gly used interchangeably and refer to a substance, Such as a cols, such as propylene glycol; (11) polyols, such as glycerin, chemical compound or complex, that has a measurable ben Sorbitol, and polyethylene glycol; (12) esters. Such eficial physiological effect on the body, such as a therapeutic 15 as ethyl oleate and ethyl laurate; (13) agar; (14) buffering effect intreatmentofadisease or disorder, when administered agents, such as hydroxide and aluminum hydrox in an effective amount. Further, when these terms are used, or ide; (15) alginic acid; (16) pyrogen-free water, (17) isotonic when a particular active agent is specifically identified by ; (18) IV fluids, including but not limited to Ringer's name or category, it is understood that Such recitation is solution, 5% dextrose in water, and half normal saline; (19) intended to include the active agent per se, as well as phar ethyl ; (20) phosphate buffer solutions; and (21) other maceutically acceptable, pharmacologically active deriva non-toxic compatible Substances employed in pharmaceuti tives thereof, or compounds significantly related thereto, cal formulations. including without limitation, salts, pharmaceutically accept “Patient” refers to animals, preferably mammals, most able salts, N-oxides, prodrugs, active metabolites, isomers, preferably humans, and includes males and females. fragments, analogs, Solvates hydrates, radioisotopes, etc. 25 “Quality of life” refers to one or more of a person’s ability The phrase “effective amount” refers to that amount of a to walk, climb stairs, do errands, work around the house, Substance that produces some desired local or systemic effect participate in recreational activities, and/or not requiring rest at a reasonable benefit/risk ratio applicable to any treatment. during the day, and/or the absence of sleeping problems or The effective amount of such substance will vary depending . upon the Subject and disease condition being treated, the 30 The term “prophylactic' or “therapeutic' treatment is art weight and age of the Subject, the severity of the disease recognized and refers to administration to the host of one or condition, the manner of administration and the like, which more of the Subject compositions. If it is administered prior to can readily be determined by one of ordinary skill in the art. clinical manifestation of the unwanted condition (e.g., dis “Endothelial dysfunction” refers to the impaired ability of ease or other unwanted state of the host animal) then the in any physiological processes carried out by the endothe 35 treatment is prophylactic, i.e., it protects the host against lium, in particular, production of nitric oxide regardless of developing the unwanted condition, whereas if administered cause. It may be evaluated by, Such as, for example, invasive after manifestation of the unwanted condition, the treatment techniques, such as, for example, coronary artery reactivity to is therapeutic (i.e., it is intended to diminish, ameliorate or acetylcholine or , and the like, or by noninvasive maintain the existing unwanted condition or side effects techniques, such as, for example, blood flow measurements, 40 brachial artery flow dilation using cuff occlusion of the arm therefrom). above or below the elbow, brachial artery ultrasonography, The term "structure-activity relationship” or “(SAR)' is imaging techniques, measurement of circulating biomarkers, art-recognized and refers to the way in which altering the Such as, asymmetric dimethylarginine (ADMA), and the like. molecular structure of a drug or other compound alters its For the latter measurement the endothelial-dependent flow 45 interaction with a receptor, enzyme, nucleic acid or other mediated dilation will be lower in patients diagnosed with an target and the like. endothelial dysfunction. It will be understood that “substitution' or “substituted The phrase “endothelial nitric oxide synthase” or “eNOS” with includes the implicit proviso that such substitution is in refers to that produce nitric oxide. accordance with permitted valence of the substituted atom The phrase “nebivolol composition” refers to a composi 50 and the substituent, and that the substitution results in a stable tion comprising nebivolol and the two are used interchange compound, e.g., which does not spontaneously undergo ably. Nebivolol is a mixture of d and 1 isomers of C.C.'- transformation Such as by rearrangement, cyclization, elimi iminobismethylene bis6-fluoro-3,4-dihydro-2H-1- nation, or other reaction. benzopyran-2-. The composition may include at The term “substituted' is also contemplated to include all least one other cardiovascular agent or at least one pharma 55 permissible Substituents of organic compounds. In a broad ceutically acceptable carrier or both. aspect, the permissible Substituents include acyclic and The term “pharmaceutically acceptable salts' is art-recog cyclic, branched and unbranched, carbocyclic and heterocy nized and refers to the relatively non-toxic, inorganic and clic, aromatic and nonaromatic Substituents of organic com organic acid addition salts of compounds, including, for pounds. Illustrative Substituents include, for example, those example, those contained in compositions of the present 60 described herein above. The permissible substituents may be invention. one or more and the same or different for appropriate organic The term “pharmaceutically acceptable carrier is art-rec compounds. For purposes of this invention, the heteroatoms ognized and refers to a pharmaceutically-acceptable material, Such as may have hydrogen Substituents and/or any composition or vehicle. Such as a liquid or Solid filler, diluent, permissible Substituents of organic compounds described excipient, solvent or encapsulating material, involved in car 65 herein which satisfy the valences of the heteroatoms. This rying or transporting any Subject composition or component invention is not intended to be limited in any manner by the thereof from one organ, or portion of the body, to another permissible Substituents of organic compounds. US 7,838,552 B2 7 8 The term “synthetic' is art-recognized and refers to pro M. Wendt M. Walter U. Geiger C. Agrawal R. Kleschyov AL, duction by chemical or enzymatic synthesis. Meinertz T. Thomas Münzel T., Arteriosclerosis, Thrombo The phrase “therapeutic effect” is art-recognized and refers sis, and Vascular Biology. 2003; 23:615-621; Troost R. to a local or systemic effect in animals, particularly mammals, Schwedhelm E. Rojczyk S, Tsikas D. Frolich J C., British and more particularly humans caused by a pharmacologically 5 Journal of Clinical Pharmacology, 2000: 50:377-379. active Substance. The term thus means any Substance Compositions Comprising Nebivolol intended for use in the diagnosis, cure, mitigation, treatment In part, the present invention features compositions com or prevention of disease or in the enhancement of desirable prising nebivolol and at least one other active agent, wherein physical or mental development and/or conditions in an ani the at least one other active agent is a cardiovascular agent. mal or human. The phrase “therapeutically-effective amount 10 The amount of each cardiovascular agent present in the com means that amount of Such a Substance that produces some positions may vary depending on a number of variables Such desired local or systemic effect at a reasonable benefit/risk as age, weight, gender, and health related issues. In general, ratio applicable to any treatment. The therapeutically effec the dosage of the cardiovascular agents will generally be in tive amount of Such substance will vary depending upon the the range of about 0.01 ng to about 10 g per kg body weight, Subject and disease condition being treated, the weight and 15 specifically in the range of about 1 ng to about 0.1 g per kg, age of the Subject, the severity of the disease condition, the and more specifically in the range of about 100 ng to about 10 manner of administration and the like, which can readily be mg per kg. In another embodiment, the amount of nebivolol in determined by one of ordinary skill in the art. the compositions of the present invention may be anywhere The term “treating is art-recognized and refers to curing as from about 0.125 mg to about 40 mg. In one example, when well as ameliorating at least one symptom of any condition or the other cardiovascular agentis an ACE inhibitor, the amount disease. of the ACE inhibitor may be anywhere from 0.5 mg to about 80 mg. When the other cardiovascular agent is an ARB, the Nebivolol amount of ARB may be anywhere from about 1 mg to about 1200 mg. The amount of the other cardiovascular agent will Nebivolol is a B-receptor blocking drug that is a mixture of 25 depend in part on the particular cardiovascular agent used. d- and 1-enantiomers, of which d-nebivolol is a highly selec In addition to ACE inhibitors and ARBs, additional cardio tive B-receptor antagonist. vascular agents include, but are not limited to adrenergic blockers, adrenergic agonists, agents for pheochromocy toma, antianginal agents, antiarrhythmics, antiplatelet 30 agents, anticoagulants, antihypertensives, antilipemic agents, (pH (pH antidiabetics, antiinflammatory agents, calcium channel s R. R. R. blockers, CETP inhibitors, COX-2 inhibitors, direct thrombin inhibitors, diuretics, endothelin receptor antagonists, HMG F F 35 Co-A reductase inhibitors, inotropic agents, rennin inhibi d-nebivolol tors, vasodilators, vasopressors, AGE crosslink breakers (ad pH pH vanced glycosylation end-product crosslink breakers, such as OnlóH-CH-NH-CH2-(H-O alagebrium, see U.S. Pat. No. 6,458.819), and AGE formation R. S S s inhibitors (advanced glycosylation end-product formation inhibitors, such as pimagedine). Cardiovascular agents fall F F 40 ing within these general categories are exemplified by the I-nebivolol following: Angiotensin I Converting Enzymes (ACEs) and Angio In addition to its B-receptor blocking properties, nebivolol tensin II Receptor Antagonists (ARBs) has been shown to cause endothelium-dependent vasodila 45 Angiotensin II receptor antagonists' (ARBs) are com tion in both normotensive and hypertensive Subjects. Cock pounds which interfere with the activity of angiotensin II by croft JR, Chowienczyk PJ, Brett SE, Chen CP. Dupont AG, binding to angiotensin II receptors and interfering with its Nueten LV. Wooding SJ, Ritter J.M., Journal of Pharmacol activity. Angiotensin I and angiotensin II are synthesized by ogy and Experimental Therapeutics. 1995; 274: 1067-1071; the enzymatic renin-angiotensin pathway. The synthetic pro TZemos N, Lim PO, MacDonald T M., Circulation, 2001; 50 cess is initiated when the enzyme renin acts on angiotensino 104:51 1-514; Broeders MA, Doevendans PA, Bekkers BC, gen, a pseudoglobulin in blood plasma, to produce the Bronsaer R, Van Gorsel E, Heemskerk J. W. Egbrink MG, Van decapeptide angiotensin I. Angiotensin I is converted by Breda E., Reneman RS, van Der Zee R., Circulation 2000; angiotensin converting enzyme (ACE) to angiotensin II (an 102:677-684. Bowman, A.J., CPL-H Chen, GA Ford. Br. J. giotensin-1-8 octapeptide). The latter is an active pressor Clin. Pharmac. 1994; 38:199-204. In experimental models, 55 Substance which has been implicated as a causative agent in nebivolol has been demonstrated to stimulate NO release several forms of hypertension in various mammalian species, through B--mediated NO production e.g., humans. and/or ATP with consequent stimulation of P2Y-pu Angiotensin II receptor antagonists (ARBs) are well rinoceptor-mediated NO release. Broeders MA, Doevendans known and include peptide compounds and non-peptide com PA, Bekkers BC, Bronsaer R, van Gorsel E, Heemskerk JW, 60 pounds. Most angiotensin II receptor antagonists are slightly Egbrink M. G. van Breda E. Reneman RS, van Der Zee R. modified congeners in which activity is attenuated by Circulation, 2000; 102:677-684; Kalinowski L., DobruckiL replacement of in position 8 with Some other W. Szczepanska-Konkel M. Jankowski M. Martyniec L. Ang ; stability can be enhanced by other replacements ielski S, Malinski T., Circulation, 2003: 107:2747-2752. It that slow degeneration in vivo. has also been reported that nebivolol inhibits NO synthase 65 Examples of angiotensin II receptor antagonists include: uncoupling and produces systemic antioxidant effects. peptidic compounds (e.g., Saralasin and related analogs); Mollnau H. Schulz, E. Daiber A, Baldus S, Oelze M. August N-substituted -2-one (U.S. Pat. No. 5,087,634); US 7,838,552 B2 10 imidazole acetate derivatives including 2-N-butyl-4-chloro 3-isopropylamino-1-(7-methylindan-4-yloxy)-2-butanol, 1-(2-chlorobenzile) imidazole-5- (see Long et al., 2-(3-t-butylamino-2-hydroxy-propylthio)-4-(5-carbamoyl J. Pharmacol. Exp. Ther. 247(1), 1-7 (1988)); 4,5,6,7-tetrahy 2-thienyl), 7-(2-hydroxy-3-t-butylaminpropoxy)ph dro-1H-imidazo 4.5-cpyridine-6-carboxylic acid and ana thalide. The above-identified compounds can be used as iso log derivatives (U.S. Pat. No. 4,816.463): N2-tetrazole beta 5 meric mixtures, or in their respective levorotating or glucuronide analogs (U.S. Pat. No. 5,085.992); substituted dextrorotating form. pyrroles, , and tryazoles (U.S. Pat. No. 5,081,127); Adrenergic Agonists and heterocyclic derivatives such as 1.3- Non-limiting examples of adrenergic agonists, both C- and (U.S. Pat. No. 5,073,566); imidazo-fused 7-member ring het B-adrenergic agonists, that may be used in the compositions erocycles (U.S. Pat. No. 5,064,825); peptides (e.g., U.S. Pat. 10 No. 4,772,684); antibodies to angiotensin II (e.g., U.S. Pat. of the present invention include , adrenalone, No. 4,302.386); and aralkyl imidazole compounds such as albuterol, , , , budrala biphenyl-methyl substituted imidazoles (e.g., EP 253,310, Zine, , , , clorprenaline, cloni Jan. 20, 1988); ES8891 (N-morpholinoacetyl-(-1-naphthyl)- dine, , , , , L-alanyl-(4, thiazolyl)-L-alanyl (35, 45)-4-amino-3-hy 15 dioxethedrine, diplivefrin, , , epineph droxy-5-cyclo-hexapentanoyl-N-hexylamide, Sankyo Com rine, , ethylnorepinephrine, , fenoxazo pany, Ltd., Tokyo, Japan); SKF 108566 (E-alpha-2-2-butyl line, , , , , 1-(carboxy phenyl)methyl1H-imidazole-5-ylmethylane hydroxyamphetamine, , indanazoline, isoetharine, 2-thiophenepropanoic acid, Smith Kline Beecham , isoproterenol, , , metaproterenol, , metizoline, , Pharmaceuticals, Pa.); Losartan (DUP753/MK954, DuPont methylhexaneamine, , , Merck Pharmaceutical Company); Remikirin (RO42-5892, , , , nor F. Hoffinan LaRoche AG); A. sub.2 agonists (Marion Merrill fenefrine, , , , oxymetazo Dow) and certain non-peptide heterocycles (G. D. Searle and line, hydrochloride, Company). Other non-limiting examples of ARBs include hydrochloride, phenylpropylmethylamine, , pir candesartan, eprosartan, irbesartan, losartan, and Valsartan. 25 Other ARBS may be identified using standard assaying tech buterol , , , . niques known to one of ordinary skill in the art. , , rillmenidine, , rito Angiotensin converting enzyme (ACE) is an enzyme drine, , Solterenol, , , terbuta which catalyzes the conversion of angiotensin I to angio line, tetrahydrozoline, , , , tensin II. ACE inhibitors include amino acids and derivatives 30 , , , , Xamot thereof, peptides, including di- and tri-peptides and antibod erol. , and mixtures thereof. ies to ACE which intervene in the renin-angiotensin system Agents for Pheochromocytoma by inhibiting the activity of ACE thereby reducing or elimi Include but are not limited to chemotherapeutics. nating the formation of pressor Substance angiotensin II. ACE inhibitors have been used medically to treat hypertension, 35 Antianginal Agents congestive heart failure, myocardial infarction and renal dis Include but are not limited to amlodipine besylate, amlo ease. Classes of compounds known to be useful as ACE dipine maleate, hydrochloride, hydro inhibitors include acylmercapto and mercaptoalkanoyl pro chloride, butoprozine hydrochloride, , lines such as captopril (U.S. Pat. No. 4,105,776) and Zofeno maleate, Succinate, , pril (U.S. Pat. No. 4.316,906), carboxyalkyl dipeptides such 40 maleate, (including but not limited to glyceryl trini as enalapril (U.S. Pat. No. 4.374,829), lisinopril (U.S. Pat. trate (GTN, nitroglycerin, Nitro-Bid), isosorbide-5-mononi No. 4.374,829), quinapril (U.S. Pat. No. 4,344.949), ramipril trate (5-ISMN, Ismo), amyl nitrate and (Icorel)), (U.S. Pat. No. 4,587.258), and perindopril (U.S. Pat. No. , hydrochloride, tosifen, 4.508,729), carboxyalkyl dipeptide mimics such as cilazapril hydrochloride). (U.S. Pat. No. 4,512,924) and benazepril (U.S. Pat. No. 4,410, 45 Antiarrhythmics 520), phosphinylalkanoyl such as fosinopril (U.S. Non-limiting examples of antiarrhythmics that may be Pat. No. 4,337.201) and trandolapril. Other non-limiting used in the compositions of the present invention include examples of ACE inhibitors include, but are not limited to, , acecainide, , , . alacepril, benazepril, captopril, ceronapril, cilaZapril, dela , amoproxan, , aprotinolol, atenolol. pril, enalapril, enalaprilat, fosinopril, imidapril, lisinopril, 50 , bevantolol, bidisomide, tosylate, bucu perindopril, quinapril, ramipril, ramiprilat, spirapril, temoca molol, butetolol, , , , pril, trandolapril. hydrochloride, butobendine, capobenic acid, , car Adrenergic Blockers teolol, cifenline, , , dolfetilide, , , , hydroquinidine, , Non-limiting examples of adrenergic blockers, both C- and 55 , , ipratropium , , B-adrenergic blockers, that may be used in the compositions , lorcainide, meobentine, , moricizine, of the present invention include Beta-adrenergic receptor , nifenaolol, , , pentisomide, blockers include, but are not limited to, atenolol, acebutolol, , , pirmenol, , prajmaline, alprenolol, , betaxolol, bunitrolol, , celip hydrochloride, , , pro rolol, hedroxalol, indenolol, , , mepin 60 dolol, methypranol, metindol, metoprolol, metrizoranolol. pranolol, pyrinoline, , sematilide, , . oXprenolol, pindolol, , practolol, Sotalolnadolol. , , , Verapamil. Viquidil, . , , timolol, bupranolol, penbutolol, trime and mixtures thereof. pranol, , 2-(3-(1,1-dimethylethyl)-amino-2-hy Antiplatelet Agents droxypropoxy)-3-pyridenecarbonitrilHCl, 1-butylamino-3- 65 Non-limiting examples of antiplatelet agents that may be (2,5-dichlorophenoxy)-2-propanol. 1-isopropylamino-3-(4- used in the compositions of the present invention include (2-cyclopropylmethoxyethyl)phenoxy)-2-propanol, , , , and . US 7,838,552 B2 11 12 Anticoagulants line, and thromboxane synthetase inhibitors Anti-coagulant agents are agents which inhibit the coagul Such as and Sulotroban, ticlopidine, , tra lation pathway by impacting negatively upon the production, pidil and ticlopidine, trifenagrel, trilinolein, 3-substituted deposition, cleavage and/or activation of factors essential in 5,6-bis(4-methoxyphenyl)-1,2,4-triazines, and antibodies to the formation of a blood clot. Non-limiting examples of anti glycoprotein IIb/IIIa as well as those disclosed in U.S. Pat. coagulants (i.e. coagulation-related therapy) that may be used No. 5,440,020, and anti- drugs, Clopidogrel; Sulfin in the compositions of the present invention include pyrazone: ; Dipyridamole; : Pyridinol Car Aggrenox, Agrylin, Amicar, Anturane, Arixtra, Coumadin, bamate; PGE: Glucagon; Antiserotonin drugs; ; Fragmin, Sodium, Lovenox, Mephyton, Miradon, : Ticlopidine. Persantine, Plavix, Pletal, Ticlid, Trental, . Other 10 “anti-coagulant” and/or “fibrinolytic' agents include Plasmi Antihypertensives nogen (to plasmin via interactions of prekallikrein, kinino Non-limiting examples of antihypertensives that may be gens, Factors XII, XIIIa, plasminogen proactivator, and tissue used in the compositions of the present invention include plasminogen activatorTPA) ; : amlodipine, benidipine, benazepril, candesartan, captopril, Anisoylated Plasminogen-Streptokinase Activator Complex; 15 , HCl, , HCl, enala Pro-Urokinase; (Pro-UK); rTPA ( or activase; r pril, eprosartan, losartan mesylate, , , denotes recombinant); rPro-UK; Abbokinase; Eminase; fosinopril, guanabenz, acetate, irbesartan, , lisino Streptase Hydrochloride; ; Dalteparin pril, , , HCl, , Sodium; Sodium; Hydrochloride: , , HCl, Efegatran ; : ; Ifetroban HCl, quinapril, , HCl, , and Sodium; ; ; Trifenagrel; War Valsartan. farin: Dextrans. This invention also contemplates fixed dose combinations Still otheranti-coagulantagents include, but are not limited of nebivolol with hydrochlorothiazide and at least one other to, ; Citrate Dextrose Solution; Antico additional active agent. agulant Citrate Phosphate Dextrose Solution; Anti 25 Antilipemic Agents coagulant Citrate Phosphate Dextrose Solution; Anticoagu Non-limiting examples of antilipemic agents that may be lant Heparin Solution; Anticoagulant Sodium Citrate used in the compositions of the present invention include Solution: Ardeparin Sodium; Bivalirudin; Bromindione: , aluminum nicotinate, atorvastatin, cholestyramine ; Desirudin; Dicumarol; Heparin Cal resin, , polidexide, beclobrate, , gemfi cium; Heparin Sodium; Lyapolate Sodium; Nafamostat 30 brozil, , lysosomal acid lipase, icofibrate, ; Mesylate; ; Tinzaparin Sodium. PPARC agonist such as , which include, but are not Inhibitors of platelet function are agents that impair the limited to , clofibrate, pirifibrate, , ability of mature platelets to perform their normal physiologi , , , theofibrate, clofibric cal roles (i.e., their normal function). Platelets are normally acid, , and ; pravastatin Sodium, simfi involved in a number of physiological processes such as 35 brate, , , nicoclonate, nicomol, oxiniacic adhesion, for example, to cellular and non-cellular entities, acid, etiroXate, thyropropic acid, thyroxine, acifran, azacos aggregation, for example, for the purpose of forming a blood terol, , beta-benzalbutyramide, carnitine, chon clot, and release of factors such as growth factors (e.g., plate droitin Sulfate clomestrone, detaXtran, dextran Sulfate let-derived growth factor (PDGF)) and platelet granular com Sodium, 5,8,11,14, 17-eicosapentaenoic acid, eritadenine, ponents. One Subcategory of platelet function inhibitors are 40 , , melinamide, my tatrienediol, , inhibitors of platelet aggregation which are compounds gamma-oryZanol, pantethine, pentaerythritol tetraacetate, which reduce or halt the ability of platelets to associate physi alpha-phenylbutyramide, pirozadil, probucol, beta-sitosterol, cally with themselves or with other cellular and non-cellular Sultosilic acid ( salt), , , xenbu components, thereby precluding the ability of a platelet to cin, and mixtures thereof. form a thrombus. 45 Examples of useful inhibitors of platelet function include Antidiabetics but are not limited to acadesine, anagrelide (if given at doses Non-limiting examples of antidiabetics that may be used in exceeding 10 mg/day), , , aspirin, clopi the compositions of the present invention include Such as , , and ; dogrel, inhibitors such as anti Such as ; derivatives Such as acetohexa inflammatory drugs and the synthetic compound FR-122047, 50 mide, 1-butyl-3-metanillylurea, , chlorpropam danaparoid sodium, daZOXiben hydrochloride, diadenosine ide, , , , , gliqui 5'5"-P1-P4-tetraphosphate () analogs, difibrotide, done, glisoxepid, glyburide, glybuthiazole, glybuzole, dihydrochloride, 1.2- and 1.3-glyceryl dinitrate, dipy glyhexamide, glymidine, glypinamide, phenbutamide, ridamole, and 3-methoxytyramine, efegatran Sul , , tolcyclamide; HDL agonists; fate, enoxaparin Sodium, glucagon, glycoprotein IIb/IIIa 55 antagonists such as Ro-43-8857 and L-700.462, ifetroban, PPARY agonists Such as Such as pioglita ifetroban Sodium, , Integrilin (), isocarba Zone, , and ; and others including cyclin methyl ester, isosorbide-5-mononitrate, itaZigrel, ket , calcium mesoxalate, , and . anserin and BM-13.177, lamifiban, lifarizine, molsidomine, Antiinflammatory Agents nifedipine, oxagrelate, PGE, platelet activating factorantago 60 Non-limiting examples of antiinflammatory agents that nists such as lexipafant, (PGI), pyrazines, pyri may be used in the compositions of the present invention dinol , ReoPro (i.e., abciximab), , include ; Dipropionate; synthetic compounds BN-50727, BN-52021, CV-4151, Acetonide; Alpha Amylase; ; ; E-5510, FK-409, GU-7, KB-2796, KBT-3022, KC-404, Sodium; Amiprilose Hydrochloride; Anakinra; KF-4939, OP-41483, TRK-100, TA-3090, TFC-612 and 65 Anirolac, ; Apazone; Balsalazide Disodium; ZK-36374, 2,4,5,7-tetrathiaoctane, 2,4,5,7-tetrathiaoctane , ; Hydrochloride: 2,2-dioxide, 2,4,5-trithiahexane, theophylline, pentoxifyl Bromelains; Broperamole; ; ; Ciclo US 7,838,552 B2 13 14 profen; Cintazone; Cliprofen; Propionate; Clobe CETP Inhibitors tasone Butyrate; Clopirac; Propionate; A non-limiting example of a CETP inhibitor that may be Cormethasone Acetate; Cortodoxone; ; Des used in the compositions of the present invention includes onide; ; Dipropionate; . Potassium; Diclofenac Sodium; Diacetate; Diflumidone Sodium; ; : COX-2 Inhibitors Diftalone; Dimethyl Sulfoxide; ; Endrysone: Non-limiting examples of COX-2 inhibitors that may be Enlimomab: Enolicam Sodium; Epirizole; .; Etofe used in the compositions of the present invention include namate; ; Fenamole: , ; Fen compounds according to the following: all of the compounds clorac; Fendosal: Fenpipalone: .; Flazalone: Fluaza 10 and substances beginning on page 8 of Winokur WO99/ cort; , ; Acetate; 20110 as members of three distinct structural classes of selec : Flunixin Meglumine: Butyl; Fluo tive COX-2 inhibitor compounds, and the compounds and rometholone Acetate; FluguaZone: ; Fluretofen; substances which are selective COX-2 inhibitors in Nicht Propionate; Furaprofen; Furobufen; Halcinon berger, U.S. Pat. No. 6,136,804, Oct. 24, 2000, entitled ide; Halobetasol Propionate; Acetate; Ibufenac: 15 "Combination therapy for treating, preventing, or reducing ; Ibuprofen Aluminum; Ibuprofen Piconol: the risks associated with acute coronary ischemic syndrome Ilonidap; Indomethacin; Indomethacin Sodium; ; and related conditions', and the compounds and Substances Indoxole; Intrazole; Isoflupredone Acetate; Isoxepac; Isoxi which are selective COX-2 inhibitors in Isakson et al., PCT cam; ; Lofemizole Hydrochloride; Lomoxicam; application WO/09641645 published Dec. 27, 1996, filed as Etabonate; Meclofenamate Sodium; Meclofe PCT/US 9509905 on Jun. 12, 1995, entitled “Combination of namic Acid; Dibutyrate; ; a Cyclooxygenase-2 Inhibitor and a Leukotriene B4 Receptor Mesalamine: MeseclaZone; Suleptan Antagonist for the Treatment of Inflammations.” The mean ate; ; ; ; Naproxen ing of COX-2 inhibitor in this invention shall include the Sodium; Naproxol; Nimazone; Olsalazine Sodium; Orgot compounds and Substances referenced and incorporated into ein; Orpanoxin: : ; Paranyline 25 Winokur WO99/20110 by reference to art therein, the com Hydrochloride; Pentosan Polysulfate Sodium; Phenbutazone pounds and Substances referenced and incorporated into Sodium Glycerate; Pirfenidone: ; Piroxicam Cin Nichtberger, U.S. Pat. No. 6,136,804, Oct. 24, 2000, by ref namate; Piroxicam Olamine; ; : Prife erence to art therein, and the compounds and Substances lone: Prodolic Acid; : Proxazole; Proxazole Cit which are COX-2 inhibitors referenced and incorporated into rate; ; Romazarit; Salcolex; Salnacedin; 30 Isakson etal, PCT application WO/09641645 published Dec. : Salicylates; Sanguinarium Chloride: SeclaZone: 27, 1996, filed as PCT/US 9509905 on Jun. 12, 1995, entitled Sermetacin; Sudoxicam, : ; Talmetacin; "Combination of a Cyclooxygenase-2 Inhibitor and a Leu Talniflumate; Talosalate; Tebufelone; : Tenidap kotriene B4 Receptor Antagonist for the Treatment of Inflam Sodium; ; Tesicam; Tesimide; Tetrydamine: mations.” The meaning of COX-2 inhibitor in this invention Tiopinac; Pivalate: Tolimetin: Tolimetin Sodium; 35 also includes , and , marketed as VIOXX ; Triflumidate; Zidometacin; ; and CELEBREX by Merck and Searle/ respectively. Sodium. One preferred antiinflammatory agent is Rofecoxib is discussed in Winokur, WO99/20110 as com aspirin. pound 3, on p. 9. Celecoxib is discussed as SC-58635 in the same reference, and in T. Penning, Synthesis and biological Calcium Channel Blockers 40 evaluation of the 1.5-diarylpyrazole class of cyclooxyge Calcium channel blockers area chemically diverse class of nase-2 inhibitors: identification of 4-5-(4-methylphenyl)-3- compounds having important therapeutic value in the control (trifluoromethyl)-1H-pyrozol-1-ylbenzenesulfonami de of a variety of diseases including several cardiovascular dis (SC-58635, celecoxib), J. Med. Chem. Apr. 25, 1997: 40(9): orders, such as hypertension, angina, and cardiac 1347-56. The meaning of COX-2 inhibitor in this invention (Fleckenstein, Cir. Res. V. 52, (suppl. 1), p. 13-16 (1983); 45 also includes SC299 referred to as a fluorescent diarylox Fleckenstein, Experimental Facts and Therapeutic Prospects, azole. C. Lanzo et al., “Fluorescence quenching analysis of the John Wiley, New York (1983); McCall, D., Curr Pract Car association and dissociation of a diarylheterocycle to diol, V. 10, p. 1-11 (1985)). Calcium channel blockers are a cyclooxygenasel-1 and cyclooxygenase-2: dynamic basis of heterogeneous group of drugs that prevent or slow the entry of cycloxygenase-2 selectivity”. Biochemistry May 23, 2000, calcium into cells by regulating cellular calcium channels. 50 vol.39(20):6228-34, and in J. Talley et al. “4,5-Diaryloxazole (Remington, The Science and Practice of Pharmacy, Nine inhibitors of cyclooxygenase-2 (COX-2), Med. Res. Rev. teenth Edition, Mack Publishing Company, Eaton, Pa., p. 963 May 1999; 19(3): 199-208. The meaning of COX-2 inhibitor (1995)). Most of the currently available calcium channel in this invention also includes . See, “4-5-Methyl blockers, and useful according to the present invention, 3-phenylisoxazol-1-ylbenzenesulfonamide, Valdecoxib: A belong to one of three major chemical groups of drugs, the 55 Potent and Selective Inhibitor of COX-2, J. Med. Chem. dihydropyridines, such as nifedipine, the phenyl alkyl 2000, Vol. 43: 775-777, and , sodium salt or pare , such as Verapamil, and the benzothiazepines, such as coxib sodium, See, N-(5-methyl-3-phenylixoxazol-4-yl)- diltiazem. Non-limiting examples of calcium channel block phenylsulfonylpropanimide, Sodium Salt, Parecoxib ers that may be used in the compositions of the present inven Sodium: A Potent and Selective Inhibitor of COX-2 for tion include , , diltiazem, , gallo 60 Parenteral Administration, J. Med. Chem. 2000, Vol. 43: pamil, mibefradil, , Semotiadil, , 1661-1663. The meaning of COX-2 inhibitor in this invention Verapamil, amlodipine, , , benidipine, also includes the Substitution of the moiety as a , , elgodipine, felodipine, isradipine, suitable replacement for the methylsulfonyl moiety. See, J. , , , nicardipine, nife Carteretal, Synthesis and activity of sulfonamide-substituted dipine, , nimodipine, nisoldipine, , 65 4.5-diaryl as selective cyclooxygenase-2 inhibi , , , , , tors.” Bioorg. Med. Chem. Lett Apr. 19, 1999: Vol. 9(8): , , , and mixtures thereof. 1171-74, and compounds referenced in the article “Design US 7,838,552 B2 15 16 and synthesis of sulfonyl-substituted 4,5-diarylthiazoles as leukin-1 alpha expression in SENCAR mouse epidermis: selective cyclooxygenase-2 inhibitors’, Bioorg. Med. Chem. implications in the prevention of stage I tumor promotion.” Lett Apr. 19, 1999: Vol. 9(8): 1167-70. The meaning of this Mol. Carcinog. December 1999, Vol 26(4):321-33 PMID invention includes a COX-2 inhibitor, NS398 referenced in 10569809. Silymarin has been used to treat diseases in two articles: Attiga et al., “Inhibitors of Synthe 5 . sis Inhibit Human Tumor Cell Invasiveness and A number of the above-identified COX-2 inhibitors are Reduce the Release of Matrix Metalloproteinases’, 60 Can prodrugs of selective COX-2 inhibitors, and exert their action cer Research 4629-4637, Aug. 15, 2000, and in “The by conversion in vivo to the active and selective COX-2 cyclooxygenase-2 inhibitor celecoxib induces apoptosis by inhibitors. The active and selective COX-2 inhibitors formed blocking Akt activation in human cells inde 10 from the above-identified COX-2 inhibitor prodrugs are pendently of Bcl-2. Hsu et al., 275(15) J. Biol. Chem. 11397 described in detail in WO95/00501, published Jan. 5, 1995, 11403 (2000). The meaning of COX-2 inhibitor in this inven WO95/18799, published Jul 13, 1995 and U.S. Pat. No. tion includes the cyclo-oxygenase-2 selective compounds 5,474,995, issued Dec. 12, 1995. Given the teachings of U.S. referenced in Mitchell et al., “Cyclo-oxygenase-2: pharma Pat. No. 5,543,297, entitled: “Human cyclooxygenase-2 cology, physiology, biochemistry and relevance to NSAID 15 cDNA and assays for evaluating cyclooxygenase-2 activity.” therapy”, Brit. J. of Pharmacology (1999) vol. 128: 1121 a person of ordinary skill in the art would be able to determine 1132, see especially p. 1126. The meaning of COX-2 inhibi whether an agent is a selective COX-2 inhibitor or a precursor tor in this invention includes so-called NO-NSAIDs or nitric of a COX-2 inhibitor, and therefore part of the present inven oxide-releasing-NSAIDs referred to in L. Jackson et al. tion. “COX-2 Selective Nonsteroidal Anti-Inflammatory Drugs: “Direct Thrombin Inhibitors’ Do They Really Offer Any Advantages?, Drugs, June, 2000 Non limiting examples of direct thrombin inhibitors vol. 59(6): 1207-1216 and the articles at footnotes 27, and 28. include , hirugen, Hirulog, argatroban, PPACK, and Also included in the meaning of COX-2 inhibitor in this thrombinaptamers. invention includes any substance that selectively inhibits the COX-2 isoenzyme over the COX-1 isoenzyme in a ratio of 25 Diuretics greater than 10 to 1 and preferably in ratio of at least 40 to 1 Non-limiting examples of diuretics that may be used in the as referenced in Winokur WO99/20110, and has one sub compositions of the present invention include althiazide, ben stituent having both atoms with free electrons under tradi droflumethiazide, benzthiazide, buthiazide, chlorthalidone, tional Valence-shell-electron-pair-repulsion theory located cyclopenthiazide, , epithiazide, ethiazide, fen on a cyclic ring (as in the Sulfylamine portion of celecoxib), 30 quizone, indapamide, hydroflumethiazide, methyclothiazide, and a second Substituent located on a different ring suffi meticrane, metolaZone, paraflutizide, polythiazide, quineth ciently far from said first substituent to have no significant aZone, teclothiazide, trichloromethiazide, chlormerodrin, electron interaction with the first substituent. The second meralluride, mercamphamide, mercaptomerin Sodium, mer substituent should have an electronegativity within such sub cumatilin Sodium, mercurous chloride, mersalyl, , stituent greater than 0.5, or the second substituent should be 35 7-morpholinomethyl-theophylline, pamabrom, protheobro an atom located on the periphery of the compound selected mine, , , oleandrin, , from the group of a halogen F, Cl, Br or I, or a group VI acetazolamide, ambuside, azosemide, , butazola element, S or O. Thus for purposes of this last included mide, clopamide, clorexolone, disulfamide, ethoXZolamide, meaning of a COX-2 inhibitor, one portion of the COX-2 , mefruside, methazolamide, , inhibitor should be hydrophilic and the other portion lipo 40 torsemide, tripamide, Xipamide, aminometradine, amisome philic. Also included as a COX-2 inhibitor are compounds tradine, amanozine, , arbutin, chlorazanil, listed at page 553 in Pharmacotherapy: A Pathophysiologic ethacrynic acid, etoZolin, hydracarbazine, isosorbide, manni Approach, Depiro et al (McGraw Hill 1999) including nabu tol, metochalcone, muZolimine, perhexiline, ticrynafen, tri metone and etodolac. Recognizing that there is overlap amterene, urea, and mixtures thereof. Depending on the among the selective COX-2 inhibitors set out in this para 45 diuretic employed, potassium may also be administered to the graph, the intent of the term COX-2 inhibitor is to compre patient in order to optimize the fluid balance while avoiding hensively include all selective COX-2 inhibitors, selective in hypokalemic alkalosis. The administration of potassium can the sense of inhibiting COX-2 over COX-1. The inventors add be in the form of potassium chloride or by the daily ingestion to the class of COX-2 inhibitors useful in the invention the of foods with high potassium content such as, for example, drug bearing the name referenced in the Wall 50 bananas or orange juice. Street Journal, Dec. 13, 2000, manufactured by Merck. See, Endothelin Receptor Antagonists also, Chauret et al., “In vitro metabolism considerations, Non-limiting examples of an endothelin receptor antago including activity testing of metabolites, in the discovery and nist that may be used in the compositions of the present selection of the COX-2 inhibitor etoricoxib (MK-0663).” invention include , Sulfonamide endothelin antago Bioorg. Med. Chem. Lett. 11 (8): 1059-62 (Apr. 23, 2001). 55 Another selective COX-2 inhibitor is DFU 5,5-dimethyl-3- nists, BQ-123, SQ28608, and the like); and mixtures thereof. (3-fluorophenyl)-4-(4-methylsulphonyl) phenyl-2(5H)-fura HMG-CoA Reductase Inhibitor (Statins) none referenced in Yergey et al. Drug Metab. Dispos. 29(5): HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) 638-44 (May 2001). The inventors also include as a selective reductase is the microsomal enzyme that catalyzes the rate COX-2 inhibitor the antioxidant silymarin, and an 60 limiting reaction in cholesterol biosynthesis (HMG active ingredient in silymarin, Silybinin, which demonstrated CoA6Mevalonate). An HMG-CoA reductase inhibitor inhib significant COX-2 inhibition relative to COX-1 inhibition. its HMG-CoA reductase, and as a result inhibits the synthesis The silymarin also showed protection against depletion of of cholesterol. A number of HMG-CoA reductase inhibitors peroxidase. Zhao et al. “Significant Inhibition by have been used to treat individuals with hypercholester the Flavonoid Antioxidant Silymarin against 12-O-tetrace 65 olemia. More recently, HMG-CoA reductase inhibitors have canoylphorbol 13-acetate-caused modulation of antioxidant been shown to be beneficial in the treatment of stroke (Endres and inflammatory enzymes, and cyclooxygenase 2 and inter M, et al., Proc Natl AcadSci USA, 1998, 95:8880-5). US 7,838,552 B2 17 18 HMG-CoA reductase inhibitors useful for co-administra flunarizine, , , isosorbide dinitrate, isosor tion with the agents of the invention include, but are not bide mononitrate, lomerixine, nafronyl, nicametate, nicer limited to, simvastatin (U.S. Pat. No. 4,444,784), lovastatin goline, nimodipine, , , tinofedrine, (U.S. Pat. No. 4.231,938), pravastatin sodium (U.S. Pat. No. Vancamine, , Viquidil, amotriphene, bendaZol. 4,346.227), fluvastatin (U.S. Pat. No. 4,739,073), atorvastatin benfurodil hemisuccinate, , chloracizine, (U.S. Pat. No. 5,273.995), cerivastatin, and numerous others chromonar, clobenfurol, clonitrate, , dilazep. described in U.S. Pat. Nos. 5,622,985; 5,135,935; 5,356,896; dipyridamole, droprenilamine, , erythrityl tetrani 4,920,109; 5,286,895; 5,262,435; 5,260,332: 5,317,031: trate, etafenone, fendiline, floredil, ganglefence, heart muscle 5,283,256; 5,256,689; 5,182,298; 5,369,125; 5,302,604; extract, bis(alpha-diethylaminoethyl ether), hex 5,166, 171; 5,202,327: 5,276,021; 5,196,440; 5,091,386; 10 obendine, hydralazine compound, itramin tosylate , 5,091,378; 4,904,646; 5,385,932; 5,250,435; 5,132,312: lidoflazine, , medibazine, nitroglycerin, 5,130,306; 5,116,870; 5,112,857; 5,102.911; 5,098,931; , isosorbide dinitrate, and other 5,081,136; 5,025,000; 5,021,453: 5,017,716; 5,001,144: nitrates, pentaerythritol tetranitrate, pentrinitrol, perhexiline, 5,001,128; 4,997,837: 4,996,234: 4,994,494; 4,992,429: pimefylline, prenylamine, propatyl nitrate, pyridofylline, tra 4,970,231: 4,968,693: 4,963,538; 4,957,940: 4,950,675; 15 pidil, tricromyl, trimetazidine, phosphate, Visna 4,946,864; 4,946,860: 4,940,800; 4,940,727: 4,939,143: dine, aluminum nicotinate, , bencyclane, betahis 4,929,620: 4,923,861; 4,906,657; 4,906,624 and 4,897,402, tine, bradykinin, brovincamine, bufeniode, , the disclosures of which patents are incorporated herein by , , , , cinnarizine, reference. , diisopropylamine dichloroacetate, eledoisin, Other non-limiting examples of HMG-CoA reductase fenoxedil, flunazine, hepronicate, ifenprodil, iloprost, inosi inhibitors that may be used in the compositions of the present tol niacinate, , kallidin, kallikrein, , invention include , , , gem nafronyl, nicametate , , nicotinyl cabene, and probucol. alcohol, nylidrin, pentifylline, pentoxifylline, , pros Inotropic Agents taglandin E1, , , niacinate, and Non-limiting examples of inotropic agents that may be 25 mixtures thereof. used in the compositions of the present invention include Note that “hydralazine compound” refers to a compound acefylline, acetyldigitoxins, 2-amino-4-picoline, anrinone, having the formula: benfurodil hemisuccinate, , camphotamide, con Vallatoxin, cymarin, denopamine, deslanoside, digitalin, digitalis, , , , docarpamine, 30 R3 R4 dopamine, dopexamine, , erythrophleine, fenal Somine, gitalin, gitoxin, glycocyamine, , hydras R FN FN FR tinine, ibopamine, lanatosides, loprinine, , nerifo lin, oleandrin, , oxyfedrine, , prenalterol, 35 whereina, b and care each independently a single or a double proscillaridin, resibufogenin, Scillaren, Scillarenin, strophan bond; R and R2 are each independently a hydrogen, an alkyl, thin, Sulmazole, theobromine, , , and an ester or a heterocyclic ring; R and Ra are each indepen mixtures thereof. dently alone pair of electrons or a hydrogen, with the proviso “Renin Inhibitors’ that at least one of R. R. R. and R is not a hydrogen. Renin inhibitors are compounds which interfere with the 40 Examples of hydralazine compounds include, but are not activity of renin. Renin inhibitors include amino acids and limited to budralazine, cadralazine, dihydralazine, endrala derivatives thereof, peptides and derivatives thereof, and anti Zine, hydralazine, pildralazine, todralazine and the like. bodies to renin. Examples of renin inhibitors that are the Vasopressors subject of United States patents are as follows: urea deriva Non-limiting examples of vasopressors that may be used in tives of peptides (U.S. Pat. No. 5,116,835); amino acids con 45 the compositions of the present invention include amezinium nected by nonpeptide bonds (U.S. Pat. No. 5,114.937); di methyl sulfate, angiotensin amide, dimetofrine, dopamine, and tri-peptide derivatives (U.S. Pat. No. 5,106,835); amino etifelmin, etillefrin, , metaraminol, methoxamine, acids and derivatives thereof (U.S. Pat. Nos. 5,104,869 and midodrine, norepinephrine, pholedrine, Synephrine, and mix 5,095,119); diol sulfonamides and sulfinyls (U.S. Pat. No. tures thereof. 5,098,924); modified peptides (U.S. Pat. No. 5,095,006): 50 peptidyl beta-aminoacylaminodiol (U.S. Pat. No. AGE Crosslink Breakers (Advanced Glycosylation End 5,089,471); pyrolimidazolones (U.S. Pat. No. 5,075,451); Product Crosslink Breakers) fluorine and chlorine statine or statone containing peptides Non-limiting examples of AGE crosslink breakers that (U.S. Pat. No. 5,066,643): peptidyl amino diols (U.S. Pat. may be used in the compositions of the present invention Nos. 5,063,208 and 4,845,079); N-morpholino derivatives 55 include Alagebrium. (U.S. Pat. No. 5,055.466); pepstatin derivatives (U.S. Pat. No. AGE Formation Inhibitors (Advanced Glycosylation End 4,980.283); N-heterocyclic (U.S. Pat. No. 4,885, Product Formation Inhibitors) 292); monoclonal antibodies to renin (U.S. Pat. No. 4,780, Non-limiting examples of AGE formation inhibitors that 401); and a variety of other peptides and analogs thereof (U.S. may be used in the compositions of the present invention Pat. Nos. 5,071,837, 5,064,965, 5,063,207, 5,036,054, 5,036, 60 include Pimagedine. 053, 5,034,512, and 4,894.437). Other Actives: Vasodilators Non-limiting examples of other active ingredients that may Non-limiting examples of vasodilators that may be used in be combined with these nebivolol compositions include, but the compositions of the present invention include bencyclane, 65 are not limited to, the following representative classes of cinnarizine, , cyclandelate, ciclonicate, diisopropy compounds, as well as their pharmaceutically acceptable lamine dichloroacetate, eburnamonine, , fenoxedil, salts, isomers, esters, ethers and other derivatives: US 7,838,552 B2 19 20 and anti-inflammatory agents, such as aloX anti-protozoal agents, such as atovaquone, benznidazole, iprin, auranofin, , benorylate, , cele , decoquinate, , coxib, diclofenac, diflunisal, etodolac, fenbufen, furoate, dinitolmide, , , nimora calcium, flurbiprofen, ibuprofen, indomethacin, ketoprofen, Zole, nitrofuraZone, and ; , leflunomide, , mefenamic acid, anti-psychotics, such as , , Ziprasi nabumetone, naproxen, oxaprozin, oxyphenbutaZone, phe done, , , , , molin nylbutaZone, piroXicam, rofecoxib, Sulindac, tetrahydrocan done, thiothixene, , , nabinol, and tromethamine; , , , , antihelminthics, such as albendazole, bephenium hydrox , , trifluopromazine, olanzap ynaphthoate, cambendazole, dichlorophen, , 10 ine; mebendazole, oxamniquine, Oxfendazole, oxantel embonate, anti-thyroid agents, such as carbimazole, paricalcitol, and praziquantel, pyrantel embonate and thiabendazole; propylthiouracil; anti- agents, such as Zileuton, , terbuta anti-tussives, such as ; line Sulfate, montelukast, and albuterol; , , hypnotics and neuroleptics, such as anti-bacterial agents. Such as alatrofloxacin, azithromycin, 15 , amylobarbitone, barbitone, , bro , benzathine , cinoxacin, mazepam, , , butobarbitone, carbro HCl, , clofazimine, cloxacillin, demeclocy mal, , chlormethiazole, chlorpromazine, cline, dirithromycin, doxycycline, , ethiona chlorprothixene, , , , cloza mide, furazolidone, grepafloxacin, , , pine, , , , fluianisone, fluni lorefloxacin, moxifloxacin HCl, nalidixic acid, nitrofuran trazepam, , flupenthixol decanoate, toin, norfloxacin, , , rifabutin, rifapentine, fluphenthixol decanoate, , , haloperi sparfloxacin, spiramycin, Sulphabenzamide, Sulphadoxine, dol, , , , , Sulphamerazine, ulphacetamide, Sulphadiazine, Sulphafura mesoridazine, , , , Zole, Sulphamethoxazole, Sulphapyridine, , tri molindone, , , , pentobarbi methoprim, trovafloxacin, and Vancomycin; 25 tone, perphenazine pimozide, , pseudoephe anti-viral agents, such as abacavir, amprenavir, delavird drine, quetiapine, risperidone, , , ine, , , lamivudine, nelfinavir, , , thioridazine, , , and Zopi , saquinavir, and stavudine; clone; anti-depressants, such as , , citalo , such as beclomethasone, , pram, , HCl, HCl, 30 budesonide, acetate, desoxymethasone, dexam HCl, HCl, HCl, ethasone, acetate, flunisolide, , HCl, HCl, maleate, and , , methylprednisolone, HC1; , and ; anti-epileptics, such as beclamide, , clon anti-parkinsonian agents, such as , bro azepam, , , Sodium, lamot 35 mocriptine meSylate, maleate, , ropin rigine, methoin, methSuximide, methylphenobarbitone, irole HCl, and : , , , phenobarbi gastrointestinal agents, such as bisacodyl, cimetidine, tone, , , , Sulthiame, tiaga , HCl, , , lan bine HCl, , Valproic acid, and vigabatrin; soprazole, , , , , 40 HCL, rabeprazole sodium, ranitidine HCl and anti-fungal agents, such as amphotericin, butenafine HCl, SulphaSalazine; nitrate, , nitrate, flu conazole, flucytosine, , , ketocona keratolytics, such as acitretin, calcipotriene, calcifediol. Zole, , , , Sulconazole nitrate, calcitriol, cholecalciferol, ergocalciferol, etretinate, retin , erbinafine HCl, , and oids, Targretin, and tazarotene; 45 lipid regulating agents, such as atorvastatin, beZafibrate, undecenoic acid; cerivastatin, ciprofibrate, clofibrate, fenofibrate, fluvastatin, anti-gout agents, such as , probenecid and Sul gemfibrozil, pravastatin, probucol, and simvastatin: phinpyrazone; muscle relaxants, such as Sodium and anti-malarials, such as amodiaquine, , chlor HC1; proguanil HCl, halofantrine HCl, HCl, proguanil 50 nutritional agents, such as calcitriol, carotenes, dihydro HCl, pyrimethamine and quinine Sulfate; tachysterol, essential fatty acids, non-essential fatty acids, anti- agents, such as mesylate, phytonadiol, A, vitamin B.Sub.2, vitamin D. Vitamin tartrate, , maleate, E and vitamin K; HCl, maleate, benzoate, analgesics, such as , , Succinate, and ; 55 diamorphine, , , , metha anti-muscarinic agents, such as , benzhexol HCl, done, , and ; , ethopropazine HCl, , sex hormones, such as clomiphene citrate, cortisone bromide, HCl and : acetate, , , ethynyl . anti-neoplastic agents and immunosuppressants, such as , fludrocortisone, , medroX , amsacrine, , , 60 yprogesterone acetate, acetate, , methylt bisantrene, buSulfan, camptothecin, capecitabine, chloram estosterone, , , oestradiol, conju bucil, cyclosporin, dacarbazine, ellipticine, , gated , , rimexolone, . etoposide, irinotecan, lomustine, melphalan, mercaptopu stilbestrol, and ; rine, , mitomycin, mitotane, mitoxantrone, , such as , dexamphetamine, mofetil mycophenolate, , , procarbazine 65 , and ; HCl, , , citrate, teniposide, tes drugs for rheumatoid arthritis Such as methotrexate, aura tolactone, topotecan HCl, and citrate; nofin, and gold sodium thiomalate; US 7,838,552 B2 21 22 drugs for osteoporosis Such as alendronate and ; col: Aphanin; Aphanol: Aphanizophyll; 8'-Apo-beta-caroten local ; 8-al; 10-Apo-beta-caroten-10'-al; 12"-Apo-beta-caroten anti-herpes drugs such as acyclovir, Valacyclovir and fam 12'al: 14'-Apo-beta-caroten-14'-al; 6'-Apo-psi-caroten-6'-al; ciclovir; 8-Apo-psi-carotein-8-al; beta-Apo-2-carotenal; beta-Apo-3- anti-emetics Such as ondansetron and ; carotenal; beta-Apo-4-carotenal; beta-Apo-2'-carotenal; and include the anthocyanidins beta-Apo-8-carotenal; beta-Apo-10'-carotenal; beta-Apo and anthocyanins; proanthocyanidins; flavan-3-ols; fla 12-carotenal; beta-Apo-14'-carotenal: Apo-8,8-caroteine Vonols; flavones; flavanones; isoflavanones; salts and esters dial; 8-Apo-beta-carotene-3,8-diol; 4'-Apo-beta-caroten-4- thereof. This development is however, not limited to fla oic acid; 8-Apo-beta-caroten-8-oic acid; 10-Apo-beta vonoid compounds isolated from plant, part of plant or 10 caroten-10'-oic acid; 12-Apo-beta-carooten-12'-oic acid; extracts of Astragalus Membranaceus, but encompasses any beta-Apo-2'-carotenoic acid; beta-Apo-2'-carotenoic acid suitable flavonoid compound isolated from different sources methylester; beta-Apo-8-carotenoic acid; beta-Apo-10'- or chemically synthesized. In addition, any suitable known or carotenoic acid; beta-Apo-12-carotenoic acid, 8-Apo-beta not yet discovered flavonoid compound, and carooten-3-ol, beta-Apo-2'-carotenol; Apo-7-fucoxanthinol; compound, is within the scope of the present technology. A 15 Apo-2-lycopenal; Apo-3-lycopenal: Apo-6' lycopenal; Apo number of flavonoids and isoflavonoids are described in 8'-lycopenal: Apo-10'-violaxanthal: Apo-12-violaxanthal; USDA-Iowa State University Database on the Isoflavone Apoviolaxanthinal; Apo-2-zeaxanthinal: Apo-3-zeaxanthi Content of Foods, Release 1.3-2002, and in USDA Database nal; Apo-4-zeaxanthinal; Astacein; Astacene; AStacene for the Flavonoid Content of Selected Foods 2003 (http:// dipalmitate; Astaxanthin; Asterinic acid; Asteroidenone; www.nal.usda.gov/fnic/foodcomp/Data/isoflav/isoflav.html) Asym. Zeta-carotene; Aurochrome; Auroxanthin; AZafirin; and (http://www.nal.usda.gov/fnic/foodcomp/Data/Flav/fla AZafrinaldehyde; v.html) (both of them herewith incorporated by reference). It Bacterial phytoene: Bacterioerythrin alpha; Bacterioeryth will be evident to any skilled person how to choose the suit rin beta; Bacteriopurpurin alpha; Bacterioruberin; alpha able flavonoid and/or isoflavonoid compound for the purpose Bacterioruberin; Bacterioruberin diglycoside: Bacterioru of the present development. For example, flavonoid com 25 berin monoglycoside; alpha-Bacterioruberin monomethyl pounds for the purpose of the present development may be, ether; Bisanhydrobacterioruberin; 3.4.3',4'-Bisdehydro-beta but are not limited to, (-)-epictechin, (+)-, procyani carotene; Bisdehydrolycopene: 2,2'-Bis(4-hydroxy-3-me din B2, dehydrate, , , and the thyl-2-butenyl)-beta.beta-carotene: 2,2'-Bis3-(glucosy like. loxy)-3-methylbutyl-3,4,3',4'-tetradehydro-1,2,1',2'- Carotenoids, generally are tetraterpenes originating from 30 tetrahydro-psi.psi-carotene-1,1'-diol; 2,2'-Bisa-(beta, D the mevalonate and deoxyxylulose phosphate pathways glucopyranosyloxy)-3-methyl-2-butenyl-gamma, gamma (older sources sometimes refer to their source as the iso carotene: 2,2'-Bis(4-hydroxy-3-methyl-2-butenyl)-gamma, prenoid pathway). Two molecules of the Co compound gera gamma-carotene; 2,2'-Bis(4-hydroxy-3-methyl-2-butenyl)- nylgeranyldiphosphate (GGDP) condense to form the sym epsilepsi-carotene: 2,2'-Bis(3-hydroxy-3-methylbutyl-3, metrical carotenoid skeleton. 35 4.3',4'-tetradehydro-1,2,1'2"tetrahydro-psi.psi-caroteine-1, Carotenoids are divided into two Subclasses, i.e., more 1'-diol; 2.2'-Bis(3-methyl-2-butenyl)-epsilepsi-carotene: polar compounds called Xanthophylls, or oxycarotenoids, and 2,2'-Bis(3-methyl-2-butenyl-3,4,3',4'-tetradehydro-1,2-di the nonpolar hydrocarbon carotenes. hydro-psi.psi-caroten-1-ol; 2.2'-Bis(3-methyl-2-butenyl)-3, Terms such as carotenoid analog and carotenoid derivative 4.3',4'-tetradehydro-1,2,1,2-tetrahydro-psi.psi-carotene-1, may generally refer to in some embodiments chemical com 40 1'-diol; 2.2'-Bis(3-methyl-2-butenyl)-1,2,1',2'-tetrahydro pounds or compositions derived from a naturally occurring psi.psi-carotene-1,1'-diol; 2,2'-Bis(O-methyl-5-C- carotenoid or simply to synthetic carotenoids. In some methylpentosyloxy)-3,4,3',4'-tetradehydro-1,2,1,2'- embodiments, terms such as carotenoid analog and caro tetrahydropsi, psi-carotene-1,1'-diol; 3,3'-Bis tenoid derivative may generally refer to chemical compounds (rhammoSyloxy)-beta,beta-carotene; 2,2'-Bis or compositions which are synthetically derived from non 45 (rhamnosyloxy)-3,4,3',4'-tetradehydro-1,2,1,2-tetrahydro carotenoid based parent compounds; however, which ulti psi, psi-carotene-1,1'-diol, Bixin; mately Substantially resemble a carotenoid derived analog. In Caloxanthin; Calthaxanthin; Canthaxanthin; Capsanthin; certain embodiments, terms such as carotenoid analog and Capsanthin epoxide; Capsanthinone; Capsanthone; Capsoch carotenoid derivative may generally refer to a synthetic rome; Capsorubin; Capsorubindione; Capsorubone; Car derivative of a naturally occurring carotenoid. Examples of 50 angoxanthin; 16'-Carboxyl-3',4'-dehydro-gamma-carotene; carotenoids are provided in the book “Carotenoids Hand Carcinoxanthin; Caricaxanthin; beta-Carotenal; psi, psi book.” edited by G. Britton et al., 2004, which is herein incor Caroten-20-al; Carotene; Caroteine X; alpha-Carotene; beta porated by reference. Carotene; beta,beta-Carotene; betagamma-Carotene; beta, It will be evident to any skilled person how to choose the epsi-Carotene; beta, phi-Carotene; beta.psi-Carotene; Suitable carotenoid compound for the purpose of the present 55 gamma-Carotene; gamma.gamma-Carotene; gamma,psi development. Carotene; delta-Carotene; epsi-Carotene; epsi (1)-Caro Examples of carotenoids include astaxanthin, Zeaxanthin, tene; epsi.epsi-Carotene; epsi.psi-Carotene; Zeta-Caro lutein, lycopene, beta-carotene. tene; Zeta-Carotene, asym.; eta-Carotene; theta-Carotene; Other non-limiting examples of naturally occurring caro Xi-Carotene; phi-Carotene; phil.phi-Carotene; phil.X- tenoids include: Aaptopurpurin; Actinioerythrin; Actinio 60 Carotene; philipsi-Carotene; XX-Carotene; psi-Carotene; erythrol; Adonirubin; Adonixanthin; A.g.470; A.g.471; Age psi.alpha-Carotene; psi.psi-Carotene; theta-Carotene; beta laxanthin C; Aleuriaxanthin; Alloxanthin; Amarouciaxanthin Carotene-5, 6.5',6'-diepoxide; beta-Carotene 5,8.5',8-diep A; Amarouciaxanthin B; Anchovy xanthin; 3',4'-Anhydrodia oxide; beta, beta-Carotene-2,2'-diol; beta,beta-Carotene-2,3- toxanthin; Anhydrodeoxyflexixanthin; Anhydroe diol; beta,beta-Carotene-3,4-diol; beta,beta-Carotene-3,3'- schscholtZXanthin; Anhydrolutein; Anhydroperidinin; Anhy 65 diol; beta.beta-Caroteine-4,4'-diol; beta.epsi-Carotene-3,2'- drorhodovibrin; Anhydrosaproxanthin; Anhydrowanningol; diol; beta.epsi-Carotene-3,3'-diol; beta.psi-Carotene-2,3- Anhydrowarmingone; Antheraxanthin; Aphanicin; Aphani diol; beta.psi-Carotene-3,3'-diol; epsi.epsi-Carotene-3,3'-

US 7,838,552 B2 35 36 Zeacarotene; beta (1)-Zeacarotene; alpha-Zeacarotene-3,17 benfurodil hemisuccinate, benidipine, benorylate, ben diol; beta-Zeacarotene-3,17-diol; beta-Zeacaroten-3-ol; tazepam, benzhexol, benziodarone, benznidazole, benzoc Zeaxanthene; Zeaxanthin: Zeaxanthin diepoxide; Zeaxanthin tamine, derivatives, benzodiazepine, benzo dimethyl ether: Zeaxanthin dirhamnoside; Zeaxanthin natate, , , beperiden, dipalmitate; Zeaxanthin 5,6-epoxide: Zeaxanthin 5.8-ep bephenium hydroxynaphthoate, bepridil, , oxide; Zeaxanthin furanoxide; Zeaxanthin monomethyl betamethasone, betaxolol, bevantolol, methylsul ether; Zeaxanthin monorhamnoside; Zeaxanthol; and Zei fate, bexarotene, bezadoxifine, bezafibrate, bialamicol, biap noxanthin. enem, bicalutamide, bietamiverine, bifonazole, binedaline, The above list of naturally occurring carotenoids is meant binifibrate, biricodar, bisacodyl, bisantrene, bisoprolol, to be a non-limiting example of naturally occurring caro 10 bitolterol, , boswellic acid, bradykinin, bretylium, tenoids. This list is not comprehensive as more naturally , , bromperidol, brotizolam, occurring molecules are being discovered which will fall brovincamine, buciclate, bucloxic acid, , budrala within the category of carotenoids. Zine, buieniode, , buflomedil, bufuliralol, bumet anide, bunitrolol, bupranolol, , bupropion, 15 , buSulfan, butalamine, , butaverine, Non-limiting examples of Sulfonylureas include, but are butenatime, butidrine hydrochloride, butobarbitone, buto not limited to , DiaBeta, , gli conazole nitrate, butoconazole, butofilol, butropium bro clazide, glipizide (Glucotrol), , chlorpropam mide, , calcifediol, calcipotriene, calcitriol, ide, tolaZamide, tolbutamide, glimepiride (Amaryl), tolbuta caldibine, cambendazole, camioxirole, camo.stat, campes mide and analogues (for example, repaglinide, terol, camptothecin, candesartan, , capecitabine, , meglitinide and (KAD-1229)) and caprate, capsaicin, captopril, carazolol, carbacephems, car the like. bamates, carbamazepine, carbapenems, carbarSone, Carba Niacin and Related Derivatives trol, , carbimazole, , carbuterol, The term niacin is the generic descriptor for nicotinic acid , carotenes, , carteolol, carvedilol. (pyridine-3-carboxylic acid) and its derivatives. Non-limiting 25 cefaclor, cefazolin, cefbuperaZone, cefepime, cefoselis, cef examples of nicotinic acid derivatives include nicofuranose, tibuten, celecoxib, , cephaeline, cephalosporin C, AcipimoX (5-methyl pyrazine-2-carboxylic acid 4-oxide), cephalosporins, cephamycins, cerivastatin, certoparin, ceta niceritrol, probucol, isonicotinic acid, Cholexamin, oxiniacic molol, cetiedil, , cetraxate, chloracizine, chloram acid, nicoclonate, nicomol, NIASPAN, nicerikol and toco bucil, chlorbetamide, chlordantoin, chlordiazepoxide, chlor pherol nicotinate. 30 madinone acetate, chlormethiazole, chloroquine, Further examples of other active agents which may be chlorothiazide, chlorpheniramine, chlorphenoxamide, chlor suitable for this invention include, without limitation: abecar , chlorproguanil, chlorpromazine, chlorpropam nil, acamprostate, acavir, acebutolol, , acemeta ide, chlorprothixene, chlortetracycline, chlorthalidone, cin, acetaminophen, acetaminosalol, , acetohexa cholecalciferol, chromonar, , ciclonicate, cido mide, maleate, acetophenazine, , 35 fovir, , , , cimetidine, acetoxypregnenolone, acetretin, acrisorcin, , acy , cinepazet maleate, , cin clovir, , adiphenine hydrochloride, adrafinil, narizine, , cinoxacin, ciprofibrate, ciprofloxacin, adrenolone, agatroban, ahnitrine, akatinol, alatrofloxacin, cisapride, cisplatin, , citicoline, clarithromycin, albendazole, albuterol, aldioxa, alendronate, , ali , , clenbuterol, clidanac, clinofibrate, bendol, alitretinoin, allopurinol, allylamines, , 40 clioquinol, clobazam, clobenfurol, , clofazimine, , , , , alprazolam, clofibrate, clofibric acid, , clomipramine, clon alprenolol, , ambucetamide, amidephrine, amidi azepam, clonidine, clonitrate, clopidogrel, clopirac nomycin, amiloride, aminoarylcarboxylic acid derivatives, indomethacin, cloranolol, cloricromen, clorprenaline, clor aminoglutethimide, , aminopentamide, ami termine, clotiazepam,: clotrimazole, cloxacillin, clozapine, nopromazine, , amiodarone, , ami 45 cmepazide, codeine methyl bromide, codeine phosphate, prilose, , , amlexanoX, amlodipine, codeine Sulfate, codeine, colloidal bismuth Subcitrate, croma amodiaquine, , amotriphene, amoxapine, amox fiban, cromolyn, cropropamide, crotethamide, , icillin, amphecloral, amphetamine, amphomycin, amphoteri cyclandelate, , , cyclexedrine, cin, amplicillin, , amprenavir, anrinone, amsa , , cyclodrine, cyclonium iodide, crine, amyl nitrate, amylobarbitone, acetate, 50 cyclopentamine, cyclosporin, cypionate, , anastroZole, andinocillin, , androstenediol-17 acetate, cytarabine, dacarbazine, dalfopristine, acetate, androstenediol-17-benzoate, androstenediol-3-ac dantrolene Sodium, , darodipine, decanoate, etate, androstenediol-3-acetate-17-benzoate, androstenedi decitabine, decoquinate, , , one, acetate, androsterone benzoate, delaviridine, demeclo cycline, denopamine, deramciclone, androsterone propionate, androsterone, angiotensin, anidula 55 descitalopram, , , 3-ke tungin, , apaZone, apicycline, apoatropine, apo todesogeskel, , desoxymethasone, detomidine, morphine, apraclonidine, aprepitant, aprotinin, arbaprostil, dexamphetamine, , , ardeparin, aripiprazole, amikacin, , arstiinol, ary dexfenfluramine, , dexraZoxane, dextro lacetic acid derivatives, arylalkylamines, arylbutyric acid amphetamine Sulfate, , dextropro derivatives, arylcarboxylic acids, arylpiperazines, arylpropi 60 poxyphene, DHEA, diacetate, diamorphine, diazemine, diaz onic acid derivatives, aspirin, , atenolol, atomox epam, diaziquinone, diazoxide, dibromopropamidine, etine, atorvastatin, atovaquone, atropine, auranofin, azapropa dichlorophen, diclofenac, , didanosine, dideoxy Zone, azathioprine, , azetazolamide, azithromycin, adenosine, diethylpropion, , , baclofen, , bamethan, barbitone, barnidipine, diflunisal, digitoxin, digoxin, dihydroergotamine, dihydroco basalazide, beclamide, beclobrate, befimolol, , 65 deine, dihydrocodeinone enol acetate, dihydroergotamine benazepril, bencyclane, bendazac, bendaZol, bendroflume mesylate, dihydroergotamine, dihydrogesterone, dihydro , benethamine penicillin, benexate hydrochloride, morphine, dihydropyridine derivatives, dihydrostreptomy US 7,838,552 B2 37 38 cin, dihydrotachysterol, dihydroxyaluminum acetylsalicy mononitrate, isosorbide dinitrate, isoxSuprine, isradipine, late, diiodohydroxyquinoline, diisopromine, dilazep. itasetron, itraconazole, itramintosylate, ivermectin, kallidin, dilevalol, diltiazem, diloxanide furoate, diloxanide, dilt kallikrein, kanamycin, , , ketoprofen, iazem, , , , dimetot ketorolac, , labetalol, , lamifiban, lamivu rine, dimorpholamine, dinitolmide, , dine, , lanatoside c, lanSoprazole, , dioxethedrine, diphemethoxidine, , diphe leflunomide, leminoprazole, lercanidipine, , letro noxylate, diphetarSone, diplivefrin, diponium bromide, dipy Zole, leucovorin, levalbuterol, , , ridamole, dirithromycin, disopyramide, divalproex sodium, levetriacetam, levobunolol, levodopa, levofloxacin, , domperidone, , dopexamine, dopradil, , , lidocaine, lidoflazine, lifi doSmalfate, , doxazosin, , , 10 brol, limaprost, , lintitript, liranaftate, lisinopril, doxycycline, drofenine, dromostanolone propionate, dromo lisuride, , lobucavir, , lomefloxacin, lom stanolone, dronabinol, droperidol, droprenilamine, d-threo erizine, lomustine, loperamide, lopinavir, , lora methylphenidate, , ebrotidine, eburnamonine, eca carbef, , lorazepam, lorefloxacin, lormetazepam, bet, ecenofloxacin, econazole nitrate, edavarone, edoxudine, losartan, lovastatin, lovastatin, loxapine Succinate, loxapine, efavirenz, effivarenZ, efloxate, eledoisin, , elgo 15 1-threo methylphenidate, , acetylsalicy dipine, ellipticine, , , enalapril, late, lysozyme, lisuride, mabuterol, mafenide, magnesium enanthate, encainide, enlopitat, enoXimone, emprostil, enta acetylsalicylate, malgramostin, mannitol hexanitrate, mapro capone, , ephedrine, , epinephrine, epiru tiline, mazindol, mebendazole, , meclofenamic bicin, , eprosartan, ergocalciferol, mesy acid, mecloxaminepentapiperide, medazepam, medibazine, lates, ergotamine, ertapenum, erythromycin, erytlirityl medigoxin, , acetate, tetranitrate, esaprazole, , esmolol, esomepra mefenamic acid, , mefloquin, mefloquine, mege Zole, esonarimod, , estradiol benzoate, estramus strol acetate, acetate, melphalan, , tine, eskiol Succinate, acetate, estrone Sulfate, mepenZolate bromide, meperidine, , etafedrine, etafenone, ethacrynic acid, ethamivan, ethi mephentermine, , , meprobamate, namate, ethinyleskadiol 3-acetate, ethinyleskadiol 3-ben 25 meptazinol, mercaptopurine, meropenum, mesalamine, Zoate, , ethionamide, (17a-ethi mesalazine, mesoridazine besylate, mesoridazine, meta nyltestosterone), ethopropazine, ethotoin, ethoxyphenamine, clazepam, , metampicillin, metaproterenol, , , ethylnorepinephrine, ethyno metaraminol, methacycline, hydrochloride, diol diacetate, etodolac, etofibrate, etoposide, etoricoxib, methadone, , methaqualone, methamphet etretinate, everolimus, exalamide, examestane, examorelin, 30 , methoin, methotrexate, methoxamine, methSuximide, eZemitibe, falecalcitriol, fanciclovir, famotidine, fantofarone, methylhexaneamine, methylphenidate d-threo-methylpheni farapenum, fargilitazar, fasudil, felbamate, felodipine, femala date, methylphenidate, methylphenobarbitone, methylpred mide, fenbuLen, , fendiline, fenfluramine, nisolone, methysergide, metiazinic acid, metizoline, meto fenoldopam, fenoprofen, fenoterol, fenoverine, fenoxazo clopramide, metolaZone, metoprolol, metoxalone, line, fenoxedil, fenpiprane, , , fentanyl. 35 metripranolol, metronidazole, mexiletine, metaxalone, , , flecainide, , floredil, mianserin, mibefradil, miconazole, midazolam, midodrine, floxuridine, fluconazole, flucytosine, fludarabine, fludiaz miglitol, , milrinone, minoxidil, , epam, fludrocortisone, flulenamic acid, flunanisone, flunariz , mitomycin, mitotane, mitoxantrone, mizolas ine, flunisolide, , fluocortolone, fluoxetine, flu tine, modafinil, , mofetil, molindone hydro penthixol decanoate, fluiphenazine decanoate, fluiphenazine 40 chloride, molindone, molsidomine, monatepil, montelukast, enanthate, fluphenazine, fluproduaZone, flurazepam, flurbi Monteplase, , moricizine, morphine hydrochloride, profen, flurogestone acetate, fluticaSone propionate, fluvas morphine Sulfate, morphine, morpholine Salicylate, tatin, , , formoterol, foScarnet, foscar , moxifloxacin, moxisylyte, moxonidine, net, fosinopril, fosphenytoin, froVatriptan, fudosteine, mycophenolate, nabumetone, , nadoxolol, nadro , furazolidone, furazolidone, furfurylmethyl 45 parin, nafamo.stat, nafronyl, , nalbuphine, nalidixic amphetamine, furosemide, gabapentin, gabexate, , acid, , , , , nan galanthamine, , gammaparin, ganciclovir, gan drolone benzoate, cyclohexanecarboxylate, nan glefene, gefarnate, gemcitabine, gemfibrozil, , ges drolone cyclohexane-propionate, , tadene, ghrelin, glatiramer, glaucarubin, glibenclamide, gli nandrolone furylpropionate, nandrolone phenpropionate, clazide, glimepiride, glipizide, gluconic acid, , 50 naphazoline, naproxen, naratriptan, natamycin, nateglinide, glyburide, glyceryl trinitrate, glimepiride, granisetron, grepa , , , nelfinavir, , floxacin, griseofulvin, guaiaZulene, guanabenz, guanfacine, undecylenate, neomycin, neokofin, , halofankine, , haloperidol, halox n-ethylamphetamine, nevirapine, nexopamil, nicametate, aZolam, hepronicate, heptanoate, , hexoprena nicardipine, nicergoline, nicofibrate, nicofuranose, nicomor line, hydramitrazine, hydrazides, , hydrocorti 55 phine, nicorandil, , nicoumalone, nifedipine, SOne, , hydroxyamphetamine, , , nilutamide, nilvadipine, nimodipine, hydroxymethylprogesterone acetate, hydroxyrnethylproges , nipradillol, nisoldipine, nitisonone, nitrazepam, terone, hydroxyprogesterone acetate, hydroxyprogesterone nitro furantoin, nitrofuraZone, nitroglycerin, nizatidine, caproate, hydroxyprogesterone, hymecromone, hyos norastemizole, norepinephrine, norethynodrel, , cyamine, ibopamine, ibudilast, ibutenac, ibuprofen, ibutilide, 60 norfloxacin, , norgeskel, , idoxuridine, ifenprodil, igmesine, iloprost, , imi , normethisterone, , norpseu dapril, imidazoles, imipenem, , , doephedrine, nortriptyline, novantrone, nylidrin, nystatin, incadronic acid , indanazoline, indenolol, indinavir, octamylamine, octodrine, octopamine, ofloxacin, olanzap indomethacin, , pranobex, inositol niaci ine, olanzapine, olapatadine, olmesartan, Olapatadine, nate, iodoquinol, ipidracine, , irbesartan, irinote 65 olsalazine, omapatrilat, omeprazole, ondansetron, , can, , isobutyrate, isocaprate esters, isoetharine, oprevelkin, , omidazole, omoprostil, oseltamivir, isometheptene, isoproterenol, isosorbide dinitrate, isosorbide oxaliplatin, oxamniquine, , Oxantel embonate, US 7,838,552 B2 39 40 oxaprozin, pemirolast, Oxatomide, oxazepam, rine, tacrine, tacrolimus, tacrolimus, , talinolol, tal oXcarbazepine, Oxfendazole, oxiconazole, , OXo ipexole, tamoxifen, , targretin, tazanolast, tazaro linic acid, oXprenolol, , oxyfedrine, oxymetazo tene, taZobactam, tecastimezole, , , line, , oxyphenbutaZone, oxyphencyclimine, , , telmisartan, temazepam, teniposide, , paclitaxel, , pantoprazole, papaverine, teprenone, teraZosin, terbinafine, terbinafine, Sul paricalcitol, paramethadione, parecoxib, pariprazole, paro fate, terbutaline, terconazole, , terodiline, terofe momycin, paroxetine, parsalmide, , , namate, , , 4-, tetra penbutolol, penciclovir, penicillin G benzathine, penicillin G cyclics, tetracycline, , tetrahydrozoline, , penicillin V, , pentaerythritol tetranitrate, , theofibrate, thiabendazole, thiazinecarboxam pentaerythritol tetranitrate, pentapiperide, pentazocine, pen 10 ides, thiocarbamates, thiocarbamizine, thiocarbarsone, thior tifylline, pentigetide, pentobarbitone, , pentoxifyl idazine, thiothixene, , tiamenidine, , line, pentrinitrol, , , pergolide, perhex , tiaramide, ticlopidine, , tilisolol. iline, perindopril erbumine, perospone, perphenazine timolol, tinidazole, tinofedrine, tinzaparin, tioconazole, pimozide, perphenazine, , phenacemide, phen tipranavir, tirapazamine, tirofiban, tiropramide, titanicene, acetin, , phencarbamide, , 15 tizanadine, tizanidine, tizinadine, tocainide, tolaZamide, tola , , , phenobarbitone, Zoline, tolbutamide, tolcapone, tolciclate, , , , phenoxybenzamine, phen , tolteridine, , tonaberstat, topiramate, Suximide, phentermine, , phenylsalicylate, phe topotecan, torsemide, toremifene cibrate, toremifene, tosu nylacetate, , phenylephrine hydrochloride, floxacin, tramadol, tramaZoline, trandolapril, tranilast, tra phenylpropanolamine hydrochloride, phenylpropanolamine nylcypromine, , traxanox, traZodone, tretoquinol, tri hydrochloride, phenylpropyl-methylamine, phenytoin, phlo acetin, triamcinolone, triamterine, , triazolam, roglucinol, pholedrine, physostigmine salicylate, physostig trifluoperazine hydrochloride, trifluoperazine, triflupro mine, phytonadiol, , piapenum, , mazine, , , , trimetazi piclaimilast, , picumast, pifarnine, pilsicainide, dine, trimethoprim, trimgestone, trimipramine, trimoprostil, pimagedine, , pimecrolimus, pimefylline, 25 trithiozine, troglitaZone, trolnibrate phosphate, pimozide, , pindolol, , pip tromethamine, tropicamide, trovafloxacin, troXipide, tuami eracillin, piperazine estrone Sulfate, piperazine derivatives, noheptane, tulobuterol, tymazoline, tyramine, undecanoate, piperilate, , piribedil, pirifibrate, piroXicam, pitav undecenoic acid, urinastatin, Valacyclovir, Valdecoxib, Valer astatin, pizotyline, plaunotol, polaprezinc, polybenzarsol, ate, Valganciclovir, Valproic acid, Valsartan, Vancomycin, polyestrol phosphate, practolol, pralnacasan, pramipexole, 30 . Venlafaxine, venorelbine, Verapamil, Vidarabine, pranlukast, pravastatin, , praziquantel, praZosin, vigabakin, , Vinpocetine, viomycin, Viquidil, vis , prenalterol, prenylamine, , prifinium bro nadine, vitamin a derivatives, vitamina, vitamin b, vitamin mide, primidone, primipramine, probenecid, probucol, d, Vitamine, Vitamink, , Voriconazole, , procainamide, procarbazine, procaterol, prochlorperazine, Xamoterol, Xanthinol niacinate, Xeny tropium bromide, proguanil, pronethalol, propafenone, , propatyl 35 Xibenolol, , Xylometazoline, yohimbine, nitrate, propentoffyline, propionate, , pro , Zafirlukast, Zalcitabine, , Zanamivir, Zate poxyphene, propranolol, propylhexedrine, propylthiouracil, bradine, , Zidovudine, Zileuton, Zimeldine, Zinc protokylol, , proxazole, pseudoephedrine, propionate, , Zolimidine, Zolmitriptan, Zolpidem, , pyrantel embonate, pyrazoles, , pyrid , , and mixtures thereof. ofylline, pyrimethamine, , pyrrolidones, 40 , quetiapine, quetuapine, , quinapril, Formulation , quinfamide, quinidine, quinine Sulfate, quinolo The nebivolol compositions of the present invention may nes, quinupritin, rabalzotan, rabeprazole sodium, rabepra be administered by various means, depending on their Zole, racefimine, ramahroban, ramipril, ranitidine, ranola intended use, as is well known in the art. For example, if Zine, ransoprazole, , rebamipide, refludan, 45 compositions of the present invention are to be administered repaglinide, , repirinast, reproterol, reserpine, retin orally, they may be formulated as tablets, capsules, granules, oids, ribavirin, rifabutine, rifampicin, rifapentine, rillmeni powders, Suspensions or syrups. Alternatively, formulations dine, , , rimiterol, rioprostil, risperidone, of the present invention may be administered parenterally as ritanovir, ritapentine, ritipenem, , ritonavir, rivastig injections (intravenous, intramuscular or Subcutaneous), drop mine, rizatriptan, , rofecoxib, rohypnol, , 50 infusion preparations or Suppositories. For application by the , ronifibrate, , , rosaprostol, ophthalmic mucous membrane route, compositions of the rosiglitaZone, rosuvastatin, rotinolol, rotraxate, roXatidine present invention may be formulated as eyedrops or eye oint acetate, , rubitecan, salacetamide, , salicyla ments. These formulations may be prepared by conventional mide, derivatives, salmeterol, saquinavir, means, and, if desired, the compositions may be mixed with saquinavir, , , , semotiadil, 55 any conventional additive, Such as an excipient, a binder, a sertindole, Sertraline, , sildenafil. Simvastatin, sir disintegrating agent, a lubricant, a corrigent, a solubilizing amesine, sirolimus, sitaxsentan, Sofalcone. Somotiadil, agent, a Suspension aid, an emulsifying agent or a coating Sorivudine, Sotalol, Soterenol, sparfloxacin, spasmolytol, agent. spectinomycin, spiramycin, Spizofurone, stavudine, Strepto In formulations of the Subject invention, wetting agents, mycin, Succinylsulfathiazole, Sucralfate, , Sulcona 60 emulsifiers and lubricants, such as Sodium lauryl Sulfate and Zole nitrate, Sulfacetamide, Sulfadiazine, Sulfaloxicacid, Sul magnesium Stearate, as well as coloring agents, release farside, , Sulindac, Suloctidil, Sulphabenzamide, agents, coating agents, Sweetening, flavoring and perfuming Sulphacetamide, Sulphadiazine, Sulphadoxine, Sulphafura agents, preservatives and antioxidants may be present in the Zole, Sulphamerazine, Sulphamethoxazole, Sulphapyridine, formulated agents. SulphaSalazine, Sulphinpyrazone, Sulpiride, Sulthiame, Sulto 65 Subject compositions may be suitable for oral, nasal, topi pride, Sulbroponium, , Sumahriptan, Sunepihon, cal (including buccal and Sublingual), rectal, vaginal, aerosol , Suplatast, Sodium, Syneph and/or parenteral administration. The formulations may con US 7,838,552 B2 41 42 Veniently be presented in unit dosage form and may be pre ethyl alcohol, , ethyl carbonate, ethyl pared by any methods well known in the art of pharmacy. The acetate, benzyl alcohol, benzyl benzoate, propylene glycol, amount of composition that may be combined with a carrier 1,3-butylene glycol, oils (in particular, cottonseed, ground material to produce a single dose vary depending upon the nut, corn, germ, olive, castor and sesame oils), glycerol, tet Subject being treated, and the particular mode of administra rahydrofuryl alcohol, polyethylene glycols and fatty. Suspen tion. sions, in addition to the Subject composition, may contain Methods of preparing these formulations include the step Suspending agents such as, for example, ethoxylated isos of bringing into association compositions of the present tearyl alcohols, polyoxyethylene Sorbitol and Sorbitan esters, invention with the carrier and, optionally, one or more acces microcrystalline cellulose, aluminum metahydroxide, bento sory ingredients. In general, the formulations are prepared by 10 nite, agar-agar and tragacanth, and mixtures thereof. uniformly and intimately bringing into association agents Formulations for rectal or vaginal administration may be with liquid carriers, or finely divided solid carriers, or both, presented as a Suppository, which may be prepared by mixing and then, if necessary, shaping the product. a subject composition with one or more suitable non-irritating Formulations suitable for may be in the excipients or carriers comprising, for example, cocoa butter, form of capsules, cachets, pills, tablets, lozenges (using a 15 polyethylene glycol, a Suppository wax or a salicylate, and flavored basis, usually Sucrose and acacia or tragacanth), which is solid at room temperature, but liquid at body tem powders, granules, or as a solution or a suspension in an perature and, therefore, will melt in the body cavity and aqueous or non-aqueous liquid, or as an oil-in-water or water release the active agent. Formulations which are suitable for in-oil liquid emulsion, or as an elixir or syrup, or as pastilles vaginal administration also include pessaries, tampons, (using an inert base, such as gelatin and glycerin, or Sucrose creams, gels, pastes, foams or spray formulations containing and acacia), each containing a predetermined amount of a Such carriers as are known in the art to be appropriate. Subject composition thereof as an active ingredient. Compo Dosage forms for transdermal administration of a subject sitions of the present invention may also be administered as a composition includes powders, sprays, ointments, pastes, bolus, electuary, or paste. creams, lotions, gels, solutions, patches and inhalants. The In Solid dosage forms for oral administration (capsules, 25 active component may be mixed understerile conditions with tablets, pills, dragées, powders, granules and the like), the a pharmaceutically acceptable carrier, and with any preser Subject composition is mixed with one or more pharmaceu vatives, buffers, or propellants which may be required. tically acceptable carriers, such as Sodium citrate or dical The ointments, pastes, creams and gels may contain, in cium phosphate, and/or any of the following: (1) fillers or addition to a Subject composition, excipients, such as animal extenders, such as starches, lactose. Sucrose, glucose, manni 30 and vegetable fats, oils, waxes, paraffins, starch, tragacanth, tol, and/or silicic acid; (2) binders, such as, for example, cellulose derivatives, polyethylene glycols, silicones, bento carboxymethylcellulose, alginates, gelatin, polyvinyl pyr nites, silicic acid, talc and Zinc oxide, or mixtures thereof. rolidone, Sucrose and/or acacia; (3) humectants, such as glyc Powders and sprays may contain, in addition to a subject erol; (4) disintegrating agents, such as agar-agar, calcium composition, excipients such as lactose, talc, silicic acid, carbonate, potato or tapioca Starch, alginic acid, certain sili 35 aluminum hydroxide, calcium silicates and polyamide pow cates, and Sodium carbonate; (5) solution retarding agents, der, or mixtures of these substances. Sprays may additionally Such as paraffin; (6) absorption accelerators, such as quater contain customary propellants, such as chlorofluorohydro nary ammonium compounds; (7) wetting agents, such as, for carbons and Volatile unsubstituted hydrocarbons, such as example, acetyl alcohol and glycerol monostearate; (8) absor and . bents, such as kaolin and bentonite clay; (9) lubricants, such 40 Compositions of the present invention may alternatively be a talc, calcium Stearate, magnesium Stearate, Solid polyethyl administered by aerosol. This is accomplished by preparing ene glycols, sodium lauryl Sulfate, and mixtures thereof, and an aqueous aerosol, liposomal preparation or solid particles (10) coloring agents. In the case of capsules, tablets and pills, containing the compound(s). A non-aqueous (e.g., fluorocar the compositions may also comprise buffering agents. Solid bon propellant) Suspension could be used. Sonic nebulizers compositions of a similar type may also be employed as fillers 45 may be used because they minimize exposing the agent to in Soft and hard-filled gelatin capsules using Such excipients shear, which may result in degradation of the compounds as lactose or milk Sugars, as well as high molecular weight contained in the Subject compositions. polyethylene glycols and the like. Ordinarily, an aqueous aerosol is made by formulating an A may be made by compression or molding, option aqueous Solution or Suspension of a Subject composition ally with one or more accessory ingredients. Compressed 50 together with conventional pharmaceutically acceptable car tablets may be prepared using binder (for example, gelatin or riers and stabilizers. The carriers and stabilizers vary with the hydroxypropyl methyl cellulose), lubricant, inert diluent, requirements of the particular subject composition, but typi preservative, disintegrant (for example, sodium starch glyco cally include non-ionic Surfactants (Tweens, Pluronics, or late or cross-linked sodium carboxymethyl cellulose), Sur polyethylene glycol), innocuous like serum albumin, face-active or dispersing agent. Molded tablets may be made 55 Sorbitan esters, oleic acid, , amino acids Such as gly by molding in a suitable machine a mixture of the Subject cine, buffers, salts, Sugars or Sugar alcohols. Aerosols gener composition moistened with an inert liquid diluent. Tablets, ally are prepared from isotonic Solutions. and other solid dosage forms, such as dragées, capsules, pills Pharmaceutical compositions of this invention suitable for and granules, may optionally be scored or prepared with parenteral administration comprise a Subject composition in coatings and shells, such as enteric coatings and other coat 60 combination with one or more pharmaceutically-acceptable ings well known in the pharmaceutical-formulating art. sterile isotonic aqueous or non-aqueous solutions, disper Liquid dosage forms for oral administration include phar sions, Suspensions or emulsions, or sterile powders which maceutically acceptable emulsions, microemulsion, Solu may be reconstituted into sterile injectable solutions or dis tions, Suspensions, syrups and elixirs. In addition to the Sub persions just prior to use, which may contain antioxidants, ject composition, the liquid dosage forms may contain inert 65 buffers, bacteriostats, solutes which render the formulation diluents commonly used in the art, Such as, for example, water isotonic with the blood of the intended recipient or suspend or other solvents, Solubilizing agents and emulsifiers, such as ing or thickening agents. US 7,838,552 B2 43 44 Examples of Suitable aqueous and non-aqueous carriers results of Such monitoring. The patient may be periodically which may be employed in the pharmaceutical compositions reevaluated to determine the extent of improvement by mea of the invention include water, , polyols (such as glyc Suring the same parameters. Adjustments to the amount(s) of erol, propylene glycol, polyethylene glycol, and the like), and Subject composition administered and possibly to the time of Suitable mixtures thereof, vegetable oils, such as olive oil, and administration may be made based on these reevaluations. injectable organic esters, such as ethyl oleate. Proper fluidity Treatment may be initiated with smaller dosages which are may be maintained, for example, by the use of coating mate less than the optimum dose of the compound. Thereafter, the rials, such as lecithin, by the maintenance of the required dosage may be increased by Small increments until the opti particle size in the case of dispersions, and by the use of mum therapeutic effect is attained. Surfactants. 10 The use of the Subject compositions may reduce the Pharmaceutical formulations may also be extended or required dosage for any individual agent contained in the delayed release formulations where the active agents are compositions (e.g., the steroidal anti inflammatory drug) released over an extended period of time. because the onset and duration of effect of the different agents Dosages may be complimentary. Administration of the compositions of the present inven 15 and therapeutic efficacy of Subject compositions tion will be in an amount sufficient to achieve a therapeutic may be determined by Standard pharmaceutical procedures in effect as recognized by one of ordinary skill in the art. cell cultures or experimental animals, e.g., for determining The dosage of any compositions of the present invention the LDso and the EDso. will vary depending on the symptoms, age and body weight of The data obtained from the cell culture assays and animal the patient, the nature and severity of the disorder to be treated studies may be used in formulating a range of dosage for use or prevented, the route of administration, and the form of the in humans. The dosage of any subject composition lies pref Subject composition. Any of the Subject formulations may be erably within a range of circulating concentrations that include the EDs with little or no toxicity. The dosage may administered in a single dose or individed doses. Dosages for vary within this range depending upon the dosage form the compositions of the present invention may be readily 25 determined by techniques known to those of skill in the art or employed and the route of administration utilized. For com as taught herein. positions of the present invention, the therapeutically effec The dosage range for nebivolol ranges from about 0.1 mg tive dose may be estimated initially from cell culture assays. In general, the doses of an active agent will be chosen by a to about 100 mg per day. In another embodiment, the dosage physician based on the age, physical condition, weight and range may be from about 0.75 mg to about 50 mg per day. In 30 yet another embodiment, the dosage range may be from about other factors known in the medical arts. 1.25 mg to about 10 mg per day. Efficacy of Treatment In certain embodiments, the dosage of the co-active com The efficacy of treatment with the subject compositions pounds will generally be in the range of about 0.01 ng to about may be determined in a number of fashions known to those of 1 g per kg body weight, specifically in the range of about 1 ng 35 skill in the art. to about 0.1 g per kg, and more specifically in the range of In one exemplary method, the median rate of decrease in about 100 ng to about 10 mg per kg body weight. inflammation for treatment with a subject composition may An effective dose or amount, and any possible affects on be compared to other forms of treatment with the particular the timing of administration of the formulation, may need to cardiovascular agent contained in the Subject composition, or be identified for any particular composition of the present 40 with other cardiovascular agents. The decrease in inflamma invention. This may be accomplished by routine experiment tion for treatment with a Subject composition as compared to as described herein, using one or more groups of animals treatment with another method may be 10, 25, 50, 75, 100, (preferably at least 5 animals per group), or in human trials if 150, 200,300, 400% greater or even more. The period of time appropriate. The effectiveness of any Subject composition for observing any such decrease may be about 1, 3, 5, 10, 15, and method of treatment or prevention may be assessed by 45 30, 60 or 90 or more hours. The comparison may be made administering the composition and assessing the effect of the against treatment with the particular cardiovascular agent administration by measuring one or more applicable indices, contained in the Subject composition, or with other cardio and comparing the post-treatment values of these indices to vascular agents, or administration of the same or different the values of the same indices prior to treatment. agents by a different method, or administration as part of a The precise time of administration and amount of any 50 different drug delivery device than a Subject composition. particular subject composition that will yield the most effec The comparison may be made against the same or a different tive treatment in a given patient will depend upon the activity, effective dosage of the various agents. pharmacokinetics, and bioavailability of a Subject composi Alternatively, a comparison of the different treatment regi tion, physiological condition of the patient (including age, mens described above may be based on the effectiveness of sex, disease type and stage, general physical condition, 55 the treatment, using standard indices known to those of skill responsiveness to a given dosage and type of ), in the art. One method of treatment may be 10%, 20%, 30%, route of administration, and the like. The guidelines presented 50%, 75%, 100%, 150%, 200%, 300% more effective, than herein may be used to optimize the treatment, e.g., determin another method. ing the optimum time and/or amount of administration, which Alternatively, the different treatment regimens may be ana will require no more than routine experimentation consisting 60 lyzed by comparing the therapeutic index for each of them, of monitoring the Subject and adjusting the dosage and/or with treatment with a subject composition as compared to timing. another regimen having atherapeutic index two, three, five or While the subject is being treated, the health of the patient seven times that of, or even one, two, three or more orders of may be monitored by measuring one or more of the relevant magnitude greater than, treatment with another method using indices at predetermined times during the treatment period. 65 Treatment, including composition, amounts, times of admin the same or different cardiovascular agents. istration and formulation, may be optimized according to the Kits US 7,838,552 B2 45 46 This invention also provides kits for conveniently and The Ca2+- complex is a cofactor for endothelial effectively implementing the methods of this invention. Such NO synthase, along with FAD, FMN, and BH4. kits comprise any Subject composition, and a means for facili Nanosensors were prepared from carbon fibers. The size of tating compliance with methods of this invention. Such kits the tip of carbon fiber was reduced from 6 um to less than 1 um provide a convenient and effective means for assuring that the by temperature controlled burning. The sensors were sensi subject to be treated takes the appropriate active in the correct tized to NO by deposition of electrically conductive poly dosage in the correct manner. The compliance means of Such kits includes any means which facilitates administering the meric porphyrin and covered with a thin layer of Nafion. The actives according to a method of this invention. Such compli porphyrinic microsensor has a response time of 0.1 ms at a ance means include instructions, packaging, and dispensing 10 micromolar NO concentration and 10 ms at the detection means, and combinations thereof. Kit components may be limit of 1 nM. packaged for either manual or partially or wholly automated The nanosensor for NO was calibrated using saturated practice of the foregoing methods. In other embodiments solution (concentration 1.82 mM verified with the coulomet involving kits, this invention contemplates a kit including ric method). Linear calibration curves were constructed for compositions of the present invention, and optionally instruc 15 each sensor from 5x10 to 3x10M NO before and after tions for their use. measurements of cell activity. The concentration-dependent effects of nebivolol and certain ACE-inhibitors on NO releas EXEMPLIFICATION ing capacity were tested using a calcium ionophore (A23 187) Example 1 that stimulates NO release, independently of G-protein coupled receptors. The data were presented as the Measurements of NO Release from Human meant-SEM for each of the triplicate measurements. The data Endothelium (calculation and plotting) were transferred to Microcal Origin 25 Software (OriginLab Corp., Northampton, Mass.). All measurements presented were recorded in vitro using a The HUVEC preparation is stable over the course of these sensitive porphyrinic probe, as previously described. Malin experiments with the cells remaining viable in culture for >24 ski T, Taha Z., Nature. 1992: 358:676-678; Malinski T, hours. Under non-stimulating conditions, basal levels of NO Czuchajowski L., Methods in Nitric Oxide Research. 1996: release are very low (<30 nM). Measurement of NO release as 319-339. NO release was measured directly from HUVEC. 30 a function of treatment was conducted in individual endothe HUVEC cells from Black and White donors were grown in Hams F12K medium with 2 mM L- adjusted to lial cells. Multiple measurements of NO release can be con contain 1.5 g/L Sodium bicarbonate and Supplemented with ducted on single cells following a brief refractory period. For 0.1 mg/ml heparin and 0.03-0.05 mg/ml endothelial cell robust statistical analysis, separate cells were used for each growth supplement (ECGS) +10% fetal bovine serum. The 35 concentration and type of drug used in these analyses. HUVEC cells were kept in an atmosphere of elevated CO2 In FIG. 1, the extent of NO release from Black and White concentration (5%). Nebivolol was obtained from Mylan donors was measured after chronic treatment with the ACE Laboratories (Morgantown, W.Va.). inhibitor, ramiprilat, followed by treatment with nebivolol (1 All measurements of endothelial NO release were con uM). At concentrations of 1, 5, and 10 uM ramiprilat, there ducted in Hank's balance solution at 37° C. Cell wells were 40 were modest but significant effects in the ability of nebivolol transferred to a Faraday cage and a porphyrinic sensor (diam to increase NO release from Black and White donor endot eter 0.5mm) was positioned at a distance of 5+2 um from the Surface of the endothelial cells using an inverted microscope helial cells. The magnitude of the increase is greater in endot (Leica Microsystems, Wetzlar, Germany) and a computer helial cells from Black donors. assisted micromanipulator. The sensor operated with a three 45 In FIG. 3, the extent of NO release from Black and White electrode system: nanosensor (working electrode), Saturated donors was measured with nebivolol (1 uM) following calomel electrode (reference electrode) and platinum wire chronic treatment with the ACE-inhibitor, enalapril. As (counter electrode, 0.5 mm diameter). The three electrodes observed with ramiprilat (above), enalapril significantly were connected to a potentiostat/galvanostat PAR273. The enhanced the ability of nebivolol to increase NO release at baseline was stabilized after about 20 seconds. The test com 50 pounds were injected with a nanoinjector onto the Surface of concentrations of 5 and 10 uM and 1, 5 and 10 uM in Black the cells following solubilization in buffer. Cells were incu and White donor cells, respectively. The magnitude of the bated with the test compounds for a 24-hour period. The increase is greater in endothelial cells from Blacks than compounds were then washed out of the system before being Whites (FIG. 4). immediately reintroduced in order to evaluate the conse 55 There were significant concentration dependent effects on quences of chronic treatment on NO release from the cells. the ability of nebivolol to enhance NO release from Black and For additive experiments, cells were incubated with ACE White donor endothelial cells that had been chronically inhibitor for 24 hours, the inhibitor was washed out of the treated with ACE inhibitors. Additionally, this property of the system, nebivolol was added and the NO release measured. drug appears to work independently of B1-adrenoceptor The current proportional to the NO concentration was mea 60 Sured with the sensor, which operated in amperometric mode blockade. By promoting a more normal vascular physiology at a constant potential of 0.63V. Data were acquired with the through an NO-dependent pathway, nebivolol treatment may use of an IBM computer with custom software and ampero have better efficacy and fewer side effects as compared to grams (current vs. time curves) were recorded with a Guniry agents that only inhibit the sympathetic . FAS 1 Femtostat (Warminster, Pa.). Maximum release of NO 65 These data further support the hypothesis that nebivolol may was produced using a calcium agonist (1 uM). By increasing cytoplasmic levels of calcium, the can bind to calmodulin. US 7,838,552 B2 47 48 have distinct pharmacologic benefits through modulation of 3. The method of claim 1 wherein the cardiovascular active endothelial function and NO metabolism. agent is olmesartan or a pharmaceutically acceptable salt thereof. INCORPORATION BY REFERENCE 4. The method of claim 1 wherein the cardiovascular active agent is Valsartan or a pharmaceutically acceptable salt All of the patents and publications cited herein are hereby thereof. incorporated by reference. 5. The method of claim 1 wherein the cardiovascular active agent is losartan or a pharmaceutically acceptable salt EQUIVALENTS thereof. 10 Those skilled in the art will recognize, or be able to ascer 6. The method of claim 2 wherein the ARB is olmesartan or tain using no more than routine experimentation, many a pharmaceutically acceptable salt thereof. equivalents to the specific embodiments of the invention 7. The method of claim 2 wherein the ARB is Valsartan or described herein. a pharmaceutically acceptable salt thereof. What is claimed is: 15 8. The method of claim 2 wherein the ARB is losartan or a 1. A method of treating hypertension comprising adminis pharmaceutically acceptable salt thereof. tering to a Subject in need thereof a composition comprising 9. A method of treating hypertension comprising adminis between about 0.125 mg to about 40 mg nebivolol or a phar tering to a Subject in need thereof a composition comprising maceutically acceptable salt thereof and between about 1 mg between about 0.125 mg to about 40 mg nebivolol or a phar to about 1200 mg of at least one other cardiovascular active maceutically acceptable salt thereof and between about 1 mg agent selected from the group consisting of Valsartan, losar to about 1200 mg of a cardiovascular active agent selected tan, olmesartan, or a pharmaceutically acceptable salt from the group consisting of Valsartan, losartan, olmesartan, thereof, wherein the effectiveness of the composition when or a pharmaceutically acceptable salt thereof wherein the administered to the Subject is greater than treatment with composition provides a higher therapeutic index than nebiv nebivolol or a cardiovascular active agent selected from the 25 olol or a cardiovascular active agent selected from the group group consisting of Valsartan, losartan, olmesartan, or a phar consisting of Valsartan, losartan, olmesartan, or a pharmaceu maceutically acceptable salt thereof administered alone. tically acceptable salt thereof administered alone. 2. A method of treating a hypertension comprising admin 10. The method of claim 9 wherein the cardiovascular istering to a Subject in need thereof a composition comprising active agent is olmesartan or a pharmaceutically acceptable between about 0.125 mg to about 40 mg nebivolol or a phar 30 maceutically acceptable salt thereof, and between about 1 mg salt thereof. to about 1200 mg of an angiotensin II receptor antagonist 11. The method of claim 9 wherein the cardiovascular (ARB), wherein the ARB is selected from the group consist active agent is Valsartan or a pharmaceutically acceptable salt ing of Valsartan, losartan, olmesartan, or a pharmaceutically thereof. 12. The method of claim 9 wherein the cardiovascular acceptable salt thereof, wherein the treatment is at least 10% 35 more effective than treatment with nebivolol or an ARB active agent is losartan or a pharmaceutically acceptable salt selected from the group consisting of Valsartan, losartan, thereof. olmesartan, or a pharmaceutically acceptable salt thereof administered alone.