US 20090226407A9 (19) United States (10) Pub. No.: US 2009/0226407 A9 (12) Patent Application Publication (48) Pub. Date: Sep. 10, 2009 L0ScalZ0 et al. CORRECTED PUBLICATION

(54) METHODS OF TREATING VASCULAR (60) Provisional application No. 60/179,020, filed on Jan. DISEASES CHARACTERIZED BY NITRC 31, 2000. Provisional application No. 60/162,230, OXIDE INSUFFICIENCY filed on Oct. 29, 1999. Provisional application No. 60/179,020, filed on Jan. 31, 2000. Provisional appli (76) Inventors: Joseph Loscalzo, Dover, MA (US); cation No. 60/162,230, filed on Oct. 29, 1999. Joseph A. Vita, Hingham, MA (US); Michael D. Loberg, Boston, MA (US); Publication Classification Manuel Worcel, Boston, MA (US) (51) Int. Cl. Correspondence Address: A6II 38/43 (2006.01) WILMERHALEANTROMED AOIN 43/04 (2006.01) 1875 PENNSYLVANIAAVE, NW (52) U.S. Cl...... 424/94.1: 514/23 WASHINGTON, DC 20006 (US) (21) Appl. No.: 10/692,724 (57) ABSTRACT (22) Filed: Oct. 27, 2003 The invention provides methods of treating and/or preventing vascular diseases characterized by nitric oxide insufficiency Prior Publication Data by administering atherapeutically effective amount of at least one nitrosated angiotensin-converting enzyme inhibitor, nit (15) Correction of US 2004/0105850 A1 Jun. 3, 2004 rosated beta-adrenergic blocker, nitrosated cholesterol reducer, nitrosated calcium channel blocker, nitrosated See (60) Related U.S. Application Data. endothelin antagonist, nitrosated angiotensin II receptor (65) US 2004/0105850 A1 Jun. 3, 2004 antagonist, nitrosated renin inhibitor, and optionally at least one compound used to treat cardiovascular diseases and/or at Related U.S. Application Data least one antioxidant, or a pharmaceutically acceptable salt thereof, and/or at least one compound that donates, transfers (60) Continuation-in-part of application No. 10/679,257. or releases nitric oxide, elevates endogenous levels of endot filed on Oct. 7, 2003, now Pat. No. 7,556,824, which is helium-derived relaxing factor, stimulates endogenous syn a division of application No. 09/697.317, filed on Oct. thesis of nitric oxide or is a substrate for nitric oxide synthase. 27, 2000, now Pat. No. 6,635,273. The antioxidant may preferably be a hydralazine compound Continuation of application No. 10/687,706, filed on or a pharmaceutically acceptable salt thereof. The compound Oct. 20, 2003, now Pat. No. 7,537,785, which is a that donates, transfers or releases nitric oxide, elevates endog continuation of application No. 10/415,136, filed on enous levels of endothelium-derived relaxing factor, stimu Apr. 25, 2003, now Pat. No. 7,235,237, filed as 371 of lates endogenous synthesis of nitric oxide or is a Substrate for international application No. PCT/US01/14245, filed nitric oxide synthase may preferably be isosorbide dinitrate on May 2, 2001, which is a continuation-in-part of and/or isosorbide mononitrate. The vascular diseases charac application No. PCT/US00/29582, filed on Oct. 26, terized by nitric oxide insufficiency include a cardiovascular 2OOO. disease and a disease resulting from oxidative stress. Patent Application Publication Sep. 10, 2009 Sheet 1 of 3 US 2009/0226407 A9

FIG. 1

Decreased Renin Release NO Insufficiency Salt/Water Retention

Increased Intracellular Increased Sensitivity to Sodium & Calcium Catecholamines and Angiotensin II

- Increased Vascul DecreasedLVH Capillary Density increasedC vascularascular Tonetone Myocardial Fibrosis

Microvascular Increased LV Mass Cardiac Ischemi Diastolic Dysfunction

-o-White hypertensives -o-Black Hypertensives

Basetne 0.3 O 3.0 10 Methachottne Dose (glnin) Patent Application Publication Sep. 10, 2009 Sheet 2 of 3 US 2009/0226407 A9 FIG. 3A

-o-White nomotensives -e-Black nomotensives gC 15 2. NE to 21 5h 5 - 41 El O PCO,05 Basettine 0.3 10 3.O. 10 Nitroprusside Dose (gfmin)

FIG. 3B -O-White nomotensives

mile- Blacknomotensives

Baseline os to so do Methachotine Dose (gfmtn) Patent Application Publication Sep.10, 2009 Sheet 3 of 3 US 2009/0226407 A9

FIG. 4A

-o- Satt Resistant (n=25) S -e - Satt Sensitive (n=6)

O - s

Basettle O.3 .0 3.0 O Methachottne Dose (g/min)

FIG. 4B

-O-Satt Resistant (n=25) -e-Satt Sensttive (n=6)

Baseline os to so to Nitroprusside Dose (glnin) US 2009/0226407 A9 Sep. 10, 2009

METHODS OF TREATING VASCULAR DISEASES that donates, transfers or releases nitric oxide, elevates endog CHARACTERIZED BY NITRC OXDE enous levels of endothelium-derived relaxing factor, stimu INSUFFICIENCY lates endogenous synthesis of nitric oxide or is a Substrate for nitric oxide synthase, and, optionally, at least one nitrosated RELATED APPLICATIONS angiotensin-converting enzyme inhibitor, nitrosated beta adrenergic blocker, nitrosated calcium channel blocker, nit 0001. This application is (i) a continuation-in-part of U.S. rosated endothelin antagonist, nitrosated angiotensin II application Ser. No. 10/679.257 filed Oct. 7, 2003, which is a receptor antagonist, nitrosated renin inhibitor, and/or at least continuation of U.S. application Ser. No. 09/697.317, filed one compound used to treat cardiovascular diseases. The Oct. 27, 2000, issued as U.S. Pat. No. 6,635,273, which present invention also provides Sustained release formulation claims priority to U.S. Provisional Application No. 60/179, comprising at least one antioxidant or a pharmaceutically 020 filed Jan. 31, 2000, and U.S. Provisional Application No. acceptable salt thereof, and at least one nitric oxide donor, 60/162.230 filed Oct. 29, 1999; and is (ii) a continuation of and, optionally, at least one nitrosated compound. U.S. Application No. 10/ filed Oct. 20, 2003, which is a continuation of U.S. application Ser. No. 10/415,136 filed BACKGROUND OF THE INVENTION Apr. 25, 2003, which is a S 371 of PCT/US01/14245 filed May 2, 2001 which claims priority to PCT/US00/29582 filed 0003. The decline in cardiovascular morbidity and mor Oct. 27, 2000. tality in the United States over the past three decades has been the result of significant advances in research on cardiovascu FIELD OF THE INVENTION lar disease mechanisms and therapeutic strategies. The inci dence and prevalence of myocardial infarction and death 0002 The present invention provides methods of treating from myocardial infarction, as well as that from cerebrovas and/or preventing vascular diseases characterized by nitric cular accident, have decreased significantly over this period oxide insufficiency by administering a therapeutically effec largely owing to advances in prevention, early diagnosis, and tive amount of at least one antioxidant or a pharmaceutically treatment of these very common diseases. acceptable salt thereof, and at least one compound that donates, transfers or releases nitric oxide, elevates endog 0004 Analysis of outcomes by race, however, paints quite enous levels of endothelium-derived relaxing factor, stimu a different picture: life expectancy and cardiovascular mor lates endogenous synthesis of nitric oxide or is a Substrate for bidity rates have improved far less for blacks than whites. nitric oxide synthase, and, optionally, at least one nitrosated Available data show that the likelihood of dying from cardio angiotensin-converting enzyme inhibitor, nitrosated beta vascular disease is far greater among black Americans than adrenergic blocker, nitrosated calcium channel blocker, nit among white Americans. In this decade, the death rate from rosated endothelin antagonist, nitrosated angiotensin II cardiovascular disease for black males was 353 per 100,000 receptor antagonist, nitrosated renin inhibitor, and/or at least population, while that for white males was 244 per 100,000; one compound used to treat cardiovascular diseases. The the rate for black females was 226 per 100,000; while that for antioxidant may preferably be a hydralazine compound or a white females was 135 per 100,000. Consonant with this pharmaceutically acceptable salt thereof. The compound that important demographic parameter is the observation that donates, transfers or releases nitric oxide, elevates endog there is a higher prevalence of several of the important risk enous levels of endothelium-derived relaxing factor, stimu factors for cardiovascular disease, e.g., hypertension, Smok lates endogenous synthesis of nitric oxide or is a Substrate for ing, diabetes mellitus, obesity, and left ventricular hypertro nitric oxide synthase may preferably be isosorbide dinitrate phy, among blacks compared with whites. In addition, out and/or isosorbide mononitrate. The present invention also comes of cardiovascular events are worse for blacks than provides methods of treating and/or preventing vascular dis whites. Following myocardial infarction, blacks have a 50% eases characterized by nitric oxide insufficiency by adminis higher annual mortality rate than whites, and their five year tering a therapeutically effective amount of at least one nit survival is only 70%. Thus, the many advances in cardiovas rosated angiotensin-converting enzyme inhibitor, nitrosated cular medicine that account for the overall improvement in beta-adrenergic blocker, nitrosated calcium channel blocker, cardiovascular health in the general population have failed to nitrosated endothelin antagonist, nitrosated angiotensin II translate into comparable racial benefits. receptor antagonist and/or nitrosated renin inhibitor, and, 0005. There is a need in the art for new and more effective optionally, at least one antioxidant and/or at least one com compositions and methods for treating vascular diseases. The pound used to treat cardiovascular diseases. The present present invention is directed to these, as well as other, impor invention also provides methods of treating and/or preventing tant ends. Raynaud's syndrome by administering a therapeutically effective amount of at least one antioxidant or a pharmaceu SUMMARY OF THE INVENTION tically acceptable salt thereof, and at least one compound that donates, transfers or releases nitric oxide, elevates endog 0006 The present invention provides methods for treating enous levels of endothelium-derived relaxing factor, stimu and/or preventing vascular diseases characterized by nitric lates endogenous synthesis of nitric oxide or is a Substrate for oxide insufficiency by administering to a patient a therapeu nitric oxide synthase, and, optionally, at least one nitrosated tically effective amount of at least one antioxidant or a phar angiotensin-converting enzyme inhibitor, nitrosated calcium maceutically acceptable salt thereof, and at least one com channel blocker, nitrosated endothelin antagonist, nitrosated pound that donates, transfers or releases nitric oxide, elevates angiotensin II receptor antagonist and/or nitrosated renin endogenous levels of endothelium-derived relaxing factor, inhibitor. The present invention also provides novel transder stimulates endogenous synthesis of nitric oxide or is a Sub mal patches comprising at least one antioxidant or a pharma strate for nitric oxide synthase, and, optionally, at least one ceutically acceptable salt thereof, and at least one compound nitrosated angiotensin-converting enzyme inhibitor, nitro US 2009/0226407 A9 Sep. 10, 2009

sated beta-adrenergic blocker, nitrosated calcium channel BRIEF DESCRIPTION OF THE FIGURES blocker, nitrosated endothelin antagonist, nitrosated angio 0012 FIG. 1 shows that nitric oxide (NO) insufficiency is tensin II receptor antagonist, nitrosated renin inhibitor, and/or associated with increased salt and water retention and a low at least one compound used to treat cardiovascular diseases. renin state. Increased intracellular sodium and calcium in The antioxidant may preferably be a hydralazine compound conjunction with reduced NO leads to enhanced sensitivity of or a pharmaceutically acceptable salt thereof. The compound vascular Smooth muscle cells and cardiomyocytes to the tonic that donates, transfers or releases nitric oxide, elevates endog and growth-stimulating properties of catecholamines and enous levels of endothelium-derived relaxing factor, stimu angiotensin II. Increased vascular tone, left ventricular hyper lates endogenous synthesis of nitric oxide or is a Substrate for trophy with inadequate capillary angiogenesis, and increased nitric oxide synthase may preferably be isosorbide dinitrate matrix production with myocardial fibrosis result. These and/or isosorbide mononitrate. The antioxidant and the nitric intermediate phenotypes lead to the clinical disorders of low oxide donor and optional nitrosated compound and/or com renin, salt-sensitive hypertension; disproportionate left ven pound used to treat cardiovascular diseases can be adminis tricular hypertrophy and diastolic dysfunction; and microvas tered separately or as components of the same composition. cular myocardial ischemia. 0007 Another aspect of the present invention provides 0013 FIG. 2 shows forearm blood flow responses to intra methods for treating and/or preventing vascular diseases arterial methacholine that were assessed using venous occlu characterized by nitric oxide insufficiency by administering sion plethysmography in 20 white and 16 black patients with to a patient a therapeutically effective amount of at least one a clinical history of hypertension (BP>140/90). FIG. 2 shows nitrosated angiotensin-converting enzyme inhibitor, nitro that endothelium-derived NO action is impaired in the fore sated beta-adrenergic blocker, nitrosated calcium channel arm microvessels of the black patients compared to the white blocker, nitrosated endothelin antagonist, nitrosated angio patients. tensin II receptor antagonist and/or nitrosated renin inhibitor, and, optionally, at least one antioxidant and/or at least one 0014 FIGS. 3A and 3B show forearm blood flow compound used to treat cardiovascular diseases. The nitro responses to nitroprusside (FIG. 3A) and methacholine (FIG. sated compound and optional antioxidant and/or compound 3B) that were assessed by venous occlusion plethysmography used to treat cardiovascular diseases can be administered in 25 white and 21 black patients without hypertension. The separately or as components of the same composition. dilator response to Sodium nitroprusside (FIG. 3A) was sig nificantly lower in black patients, while there was no racial 0008. In another aspect, the present invention provides difference in response to methacholine (FIG. 3B). methods for treating and/or preventing Raynaud's syndrome by administering to a patient a therapeutically effective 0.015 FIGS. 4A and 4B show the effect of salt-sensitivity amount of at least one antioxidant or a pharmaceutically on forearm microvascular function. By repeated measures acceptable salt thereof, and at least one compound that ANOVA, there were trends for impaired responses to metha donates, transfers or releases nitric oxide, elevates endog choline (FIG. 4A) and sodium nitroprusside (FIG. 4B) in enous levels of endothelium-derived relaxing factor, stimu salt-sensitive black patients. lates endogenous synthesis of nitric oxide or is a Substrate for nitric oxide synthase, and, optionally, at least one nitrosated DETAILED DESCRIPTION OF THE INVENTION angiotensin-converting enzyme inhibitor, nitrosated calcium 0016. As used throughout the disclosure, the following channel blocker, nitrosated endothelin antagonist, nitrosated terms, unless otherwise indicated, shall be understood to have angiotensin II receptor antagonist and/or nitrosated renin the following meanings. inhibitor. The antioxidant, nitric oxide donor, and nitrosated compound can be administered separately or as components 0017 “Patient” refers to animals, preferably mammals, of the same composition. most preferably humans, and includes males and females. 0018 “Therapeutically effective amount” refers to the 0009. In yet another aspect, the present invention provides amount of the compound and/or composition that is effective novel transdermal patches comprising atherapeutically effec to achieve its intended purpose. tive amount of at least one antioxidant at least one compound that donates, transfers or releases nitric oxide, elevates endog 0.019 “Transdermal” refers to the delivery of a compound enous levels of endothelium-derived relaxing factor, stimu by passage through the skin and into the blood stream. lates endogenous synthesis of nitric oxide or is a Substrate for 0020 “Transmucosal” refers to delivery of a compound by nitric oxide synthase, and, optionally, at least one nitrosated passage of the compound through the mucosal tissue and into angiotensin-converting enzyme inhibitor, nitrosated beta the blood stream. adrenergic blocker, nitrosated calcium channel blocker, nit rosated endothelin antagonist, nitrosated angiotensin II 0021 “Penetration enhancement' or “permeation receptor antagonist, nitrosated renin inhibitor, and/or at least enhancement” refers to an increase in the permeability of the one compound used to treat cardiovascular diseases. skin or mucosal tissue to a selected pharmacologically active compound Such that the rate at which the compound perme 0010. In another aspect, the present invention provides ates through the skin or mucosal tissue is increased. Sustained release formulations comprising a therapeutically effective amount of at least one antioxidant or a pharmaceu 0022. “Carriers” or “vehicles” refers to carrier materials tically acceptable Salt thereof, and at least one nitric oxide Suitable for compound administration and include any Such donor, and, optionally, at least one nitrosated compound. material known in the art such as, for example, any liquid, gel. solvent, liquid diluent, solubilizer, or the like, which is non 0011. These and other aspects of the present invention are toxic and which does not interact with any components of the described in more detail herein. composition in a deleterious manner. US 2009/0226407 A9 Sep. 10, 2009

0023 “Sustained release' refers to the release of a thera thereof fused via adjacent or non-adjacent atoms. Bridged peutically active compound and/or composition Such that the cycloalkyl groups can be unsubstituted or Substituted with blood levels of the therapeutically active compound are main one, two or three substituents independently selected from tained within a desirable therapeutic range over an extended alkyl, alkoxy, amino, alkylamino, dialkylamino, hydroxy, period of time. The sustained release formulation can be halo, carboxyl, alkylcarboxylic acid, aryl, amidyl, ester, alky prepared using any conventional method knownto one skilled lcarboxylic ester, carboxamido, alkylcarboxamido, oxo and in the art to obtain the desired release characteristics. nitro. Exemplary bridged cycloalkyl groups include adaman 0024 “Nitric oxide donor” or “NO donor” refers to com tyl, decahydronapthyl, quinuclidyl, 2,6-dioxabicyclo3.3.0 pounds that donate, release and/or directly or indirectly trans octane, 7-Oxabycyclo2.2.1]heptyl, 8-azabicyclo3.2.1]oct fera nitrogen monoxide species, and/or stimulate the endog 2-enyl and the like. enous production of nitric oxide or endothelium-derived 0032 “Cycloalkyl refers to a saturated or unsaturated relaxing factor (EDRF) in vivo and/or elevate endogenous cyclic hydrocarbon comprising from about 3 to about 10 levels of nitric oxide or EDRF in vivo. “NO donor also carbon atoms. Cycloalkyl groups can be unsubstituted or includes compounds that are Substrates for nitric oxide Syn substituted with one, two or three substituents independently thase. selected from alkyl, alkoxy, amino, alkylamino, dialky 0025 “Nitric oxide adduct” or “NO adduct” refers to com lamino, arylamino, diarylamino, alkylarylamino, aryl, pounds and functional groups which, under physiological amidyl, ester, hydroxy, halo, carboxyl, alkylcarboxylic acid, conditions, can donate, release and/or directly or indirectly alkylcarboxylic ester, carboxamido, alkylcarboxamido, oxo transfer any of the three redox forms of nitrogen monoxide and nitro. Exemplary cycloalkyl groups include cyclopropyl. (NO", No, NO), such that the biological activity of the cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cyclo nitrogen monoxide species is expressed at the intended site of hepta, 1,3-dienyl, and the like. action. 0033 “Heterocyclic ring or group' refers to a saturated, 0026 “Nitric oxide releasing or “nitric oxide donating unsaturated, cyclic or aromatic or polycyclic hydrocarbon refers to methods of donating, releasing and/or directly or group having about 2 to about 10 carbon atoms (preferably indirectly transferring any of the three redox forms of nitro about 4 to about 6 carbon atoms) where 1 to about 4 carbon gen monoxide (NO", NO-, NO), such that the biological atoms are replaced by one or more nitrogen, oxygen and/or activity of the nitrogen monoxide species is expressed at the sulfur atoms. Sulfur may be in the thio, sulfinyl or sulfonyl intended site of action. oxidation state. The heterocyclic ring or group can be fused to an aromatic hydrocarbon group. Heterocyclic groups can be 0027 “Alkyl refers to a lower alkyl group, a haloalkyl unsubstituted or substituted with one, two or three substitu group, an alkenyl group, an alkynyl group, a bridged ents independently selected from alkyl, alkoxy, amino, alky cycloalkyl group, a cycloalkyl group or a heterocyclic ring, as lamino, dialkylamino, arylamino, diarylamino, alkylary defined herein. lamino, hydroxy, oxo, thial, halo, carboxyl, carboxylic ester, 0028 “Lower alkyl refers to branched or straight chain alkylcarboxylic acid, alkylcarboxylic ester, aryl, arylcar acyclic alkyl group comprising one to about ten carbonatoms boxylic acid, arylcarboxylic ester, amidyl, ester, carboxa (preferably one to about eight carbon atoms, more preferably mido, alkylcarboxamido, arylcarboxamido, Sulfonic acid, one to about six carbonatoms). Exemplary lower alkyl groups Sulfonic ester, Sulfonamido and nitro. Exemplary heterocy include methyl, ethyl, n-propyl, isopropyl. n-butyl, isobutyl, clic groups include pyrrolyl, 3-pyrrolinyl, 4.5,6-trihydro-2H sec-butyl, t-butyl, pentyl, neopentyl, iso-amyl, hexyl, octyl, pyranyl, pyridinyl, 1,4-dihydropyridinyl, pyrazolyl, triazolyl, and the like. pyrimidinyl, pyridazinyl, oxazolyl, thiazolyl, imidazolyl, 0029) “Haloalkyl refers to a lower alkyl group, an alkenyl indolyl, thiophenyl, furanyl, tetrhydrofuranyl, tetrazolyl, group, an alkynyl group, a bridged cycloalkyl group, a 2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl, oxazolindinyl 1.3- cycloalkyl group or a heterocyclic ring, as defined herein, to dioxolanyl, 2,6-dioxabicyclo3.3.0octanyl, 2-imidazonli which is appended one or more halogens, as defined herein. nyl, imidazolindinyl, 2-pyrazolinyl, pyrazolidinyl, isox Exemplary haloalkyl groups include trifluoromethyl, chlo azolyl, isothiazolyl, 1.2.3-oxadiazolyl, 1.2.3-triazolyl, 1.3,4- romethyl, 2-bromobutyl, 1-bromo-2-chloro-pentyl, and the thiadiazolyl. 2H-pyranyl, 4H-pyranyl, piperidinyl, 1,4- like. “Alkenyl refers to a branched or straight chain C-Co dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl, hydrocarbon (preferably a C-C hydrocarbon, more prefer pyrazinyl, piperazinyl, 1.3.5-triazinyl, 1,3,5-trithianyl, ben ably a C-C hydrocarbon) which can comprise one or more ZO(b)thiophenyl, benzimidazolyl, quinolinyl, and the like. carbon-carbon double bonds. Exemplary alkenyl groups 0034) “Heterocyclic compounds’ refer to mono- and poly include propylenyl, buten-1-yl, isobutenyl, penten-1-yl, 2.2- cyclic compounds comprising at least one aryl or heterocyclic methylbuten-1-yl, 3-methylbuten-1-yl, hexan-1-yl, hepten ring. 1-yl, octen-1-yl, and the like. 0035 "Aryl refers to a monocyclic, bicyclic, carbocyclic 0030) “Alkynyl” refers to an unsaturated acyclic C-Co or heterocyclic ring system comprising one or two aromatic hydrocarbon (preferably a C-Cs hydrocarbon, more prefer rings. Exemplary aryl groups include phenyl, pyridyl, ably a C-C hydrocarbon) which can comprise one or more napthyl, quinoyl, tetrahydronaphthyl, furanyl, indanyl, inde carbon-carbon triple bonds. Exemplary alkynyl groups nyl, indoyl, and the like. Aryl groups (including bicylic aryl include ethynyl, propynyl, butyn-1-yl, butyn-2-yl, pentyl-1- groups) can be unsubstituted or Substituted with one, two or yl, pentyl-2-yl, 3-methylbutyn-1-yl, hexyl-1-yl, hexyl-2-yl, three substituents independently selected from alkyl, alkoxy, hexyl-3-yl, 3.3-dimethylbutyn-1-yl, and the like. amino, alkylamino, dialkylamino, arylamino, diarylamino, 0031) “Bridged cycloalkyl” refers to two or more alkylarylamino, hydroxy, carboxyl, carboxylic ester, alkyl cycloalkyl groups, heterocyclic groups, or a combination carboxylic acid, alkylcarboxylic ester, aryl, arylcarboxylic US 2009/0226407 A9 Sep. 10, 2009 acid, arylcarboxylic ester, alkylcarbonyl, arylcarbonyl, 0051) “Oxy” refers to - O TR," wherein R, is an amidyl, ester, carboxamido, alkylcarboxamido, carbomyl. organic or inorganic cation. Sulfonic acid, Sulfonic ester, Sulfonamido and nitro. Exem plary Substituted aryl groups include tetrafluorophenyl, pen 0052 “Organic cation” refers to a positively charged tafluorophenyl, Sulfonamide, alkylsulfonyl, arylsulfonyl, and organic ion. Exemplary organic cations include alkyl Substi the like. tuted ammonium cations, and the like. 0.036 “Alkylaryl refers to an alkyl group, as defined 0053 “Inorganic cation” refers to a positively charged herein, to which is appended an aryl group, as defined herein. metalion. Exemplary inorganic cations include Group I metal Exemplary alkylaryl groups include benzyl, phenylethyl, cations such as for example, Sodium, potassium, and the like. hydroxybenzyl, fluorobenzyl, fluorophenylethyl, and the 0054) “Hydroxyalkyl refers to a hydroxy group, as like. defined herein, appended to an alkyl group, as defined herein. 0037 'Arylalkyl refers to an aryl radical, as defined 0.055 “Amino” refers to -NH. herein, attached to an alkyl radical, as defined herein. 0056 “Nitrate” refers to - O NO. 0038 “Cycloalkylalkyl refers to a cycloalkyl radical, as defined herein, attached to an alkyl radical, as defined herein. 0057. “Nitrite” refers to O NO. 0039) “Heterocyclicalkyl” refers to a heterocyclic ring 0.058 “Thionitrate” refers to S NO. radical, as defined herein, attached to an alkyl radical, as 0059) “Thionitrite” and “nitrosothiol” refer to S NO. defined herein. 0060 “Nitro” refers to the group - NO, and “nitrosated” 0040 “Cycloalkenyl refers to an unsaturated cyclic refers to compounds that have been substituted therewith. hydrocarbon having about 3 to about 10 carbon atoms (pref 0061 “Nitroso’ refers to the group - NO and “nitrosy erably about 3 to about 8 carbonatoms, more preferably about lated refers to compounds that have been substituted there 3 to about 6 carbon atoms) comprising one or more carbon with. carbon double bonds. 0041 Arylheterocyclic ring refers to a bi- or tricyclic 0062) “Nitrile' and “cyano” refer to CN. ring comprised of an aryl ring, as defined herein, appended 0063) “Halogen' or “halo” refers to iodine (I), bromine via two adjacent carbon atoms of the aryl ring to a heterocy (Br), chlorine (Cl), and/or fluorine (F). clic ring, as defined herein. Exemplary arylheterocyclic rings 0064 Alkylamino” refers to RsNH , wherein Rs is an include dihydroindole, 1,2,3,4-tetra-hydroquinoline, and the alkyl group, as defined herein. Exemplary alkylamino groups like. include methylamino, ethylamino, butylamino, cyclohexy 0.042 Alkoxy' refers to Rs.O. , wherein Rs is an alkyl lamino, and the like. group, as defined herein. Exemplary alkoxy groups include 0065 'Arylamino” refers to RsNH , wherein Rss is an methoxy, ethoxy, t-butoxy, cyclopentyloxy, and the like. aryl group, as defined herein. 0.043 Arylalkoxy or alkoxyaryl” refers to an alkoxy 0066 “Dialkylamino” refers to RsRN , wherein Rso group, as defined herein, to which is appended an aryl group, and Rs are each independently an alkyl group, as defined as defined herein. Exemplary arylalkoxy groups include ben herein. Exemplary dialkylamino groups include dimethy Zyloxy, phenylethoxy, chlorophenylethoxy, and the like. lamino, diethylamino, methyl propargylamino, and the like. 0044 "Aryloxy” refers to Rs 0 , wherein Rss is an aryl 0067 "Diarylamino” refers to RssRN , wherein Rss group, as defined herein. Exemplary aryloxy groups include and Reo are each independently an aryl group, as defined napthyloxy, quinolyloxy, isoquinolizinyloxy, and the like. herein. 0045 “Alkoxyalkyl refers to an alkoxy group, as defined herein, appended to an alkyl group, as defined herein. Exem 0068 Alkylarylamino” refers to RsRsN , wherein plary alkoxyalkyl groups include methoxymethyl, methoxy R is an alkyl group, as defined herein, and Rss is an aryl ethyl, isopropoxymethyl, and the like. group, as defined herein. 0069 Aminoalkyl refers to an amino group, an alky 0046 “Alkoxyhaloalkyl refers to an alkoxy group, as lamino group, a dialkylamino group, an arylamino group, a defined herein, appended to a haloalkyl group, as defined diarylamino group, an alkylarylamino group or a heterocyclic herein. Exemplary alkoxyhaloalkyl groups include 4-meth ring, as defined herein, to which is appended an alkyl group, oxy-2-chlorobutyl and the like. as defined herein. 0047 “Cycloalkoxy' refers to Rs 0 , wherein Rs is a 0070 Aminoaryl refers to an amino group, an alky cycloalkyl group or a bridged cycloalkyl group, as defined lamino group, a dialkylamino group, an arylamino group, a herein. Exemplary cycloalkoxy groups include cyclopropy diarylamino group, an alkylarylamino group or a heterocyclic loxy, cyclopentyloxy, cyclohexyloxy, and the like. ring, as defined herein, to which is appended an aryl group, as 0.048 “Haloalkoxy' refers to a haloalkyl group, as defined defined herein. herein, to which is appended an alkoxy group, as defined herein. Exemplary haloalkyl groups include 1,1,1-trichloro 0.071) “Thio” refers to - S - ethoxy, 2-bromobutoxy, and the like. 0072 "Sulfinyl refers to S(O)-. 0049) “Hydroxy” refers to OH. 0.073 "Methanthial' refers to C(S)-. 0050 “Oxo” refers to —O. 0.074) “Thial” refers to =S. US 2009/0226407 A9 Sep. 10, 2009

0075) “Sulfonyl refers to S(O). heterocyclic ring, as defined herein, or Rs and Rs 7 taken together are a heterocyclic ring, a cycloalkyl group or a 0076 “Sulfonic acid” refers to S(O),OR, wherein R is a hydrogen, an organic cation or an inorganic cation. bridged cycloalkyl group, as defined herein. 0.077 Alkylsulfonic acid refers to a sulfonic acid group, 0094) “Carboxyl refers to C(O)CR, wherein R is a as defined herein, appended to an alkyl group, as defined hydrogen, an organic cation oran inorganic cation, as defined herein. herein. 0078 “Arylsulfonic acid refers to an sulfonic acid group, 0095 “Carbonyl refers to C(O)-. as defined herein, appended to an aryl group, as defined herein 0.096 “Alkylcarbonyl' or “alkanoyl refers to Rs 0079) “Sulfonic ester” refers to S(O)ORs, wherein C(O)—, wherein Rs is an alkyl group, as defined herein. R is an alkyl group, an aryl group, an alkylaryl group or an arylheterocyclic ring, as defined herein. 0097) 'Arylcarbonyl' or “aroyl” refers to Rss C(O) , wherein Rss is an aryl group, as defined herein. 0080) “Sulfonamido” refers to S(O), N(Rs)(Rs), wherein Rs, and Rs, are each independently a hydrogenatom, 0.098 “Carboxylic ester” refers to C(O)ORs, wherein an alkyl group, an aryl group, an alkylaryl group, or an aryl R is an alkyl group, an aryl group, an alkylaryl group or an heterocyclic ring, as defined herein, or Rs, and Rs, taken arylheterocyclic ring, as defined herein. together are a heterocyclic ring, a cycloalkyl group or a 0099) “Alkylcarboxylic acid” and “alkylcarboxyl refer to bridged cycloalkyl group, as defined herein. an alkyl group, as defined herein, appended to a carboxyl 0081 “Alkylsulfonamido” refers to a sulfonamido group, group, as defined herein. as defined herein, appended to an alkyl group, as defined 0.100 Alkylcarboxylic ester refers to an alkyl group, as herein. defined herein, appended to a carboxylic ester group, as 0082 "Arylsulfonamido” refers to a sulfonamido group, defined herein. as defined herein, appended to an aryl group, as defined herein. 0101 Arylcarboxylic acid refers to an aryl group, as defined herein, appended to a carboxyl group, as defined 0083) “Alkylthio” refers to RS , wherein Rs is an herein. alkyl group, as defined herein. 0102) “Arylcarboxylic ester” and “arylcarboxyl refer to 0084 "Arylthio” refers to RS , wherein Rss is an aryl an aryl group, as defined herein, appended to a carboxylic group, as defined herein. ester group, as defined herein. 0085 “Cycloalkylthio” refers to RS , wherein Rs is a 0103). “Carboxamido” refers to C(O)N(Rs)(Rs), cycloalkyl group or a bridged cycloalkyl group, as defined wherein Rs, and Rs, are each independently a hydrogenatom, herein. Exemplary cycloalkylthio groups include cyclopro an alkyl group, an aryl group, an alkylaryl group or an aryl pylthio, cyclopentylthio, cyclohexylthio, and the like. heterocyclic ring, as defined herein, or Rs, and Rs, taken I0086 “Alkylsulfinyl" refers to Rs S(O)—, wherein Rso together with the nitrogen to which they are attached form a is an alkyl group, as defined herein. heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein. 0087) “Alkylsulfonyl refers to Rs. S(O) , wherein R is an alkyl group, as defined herein. 0.104 Alkylcarboxamido” refers to an alkyl group, as defined herein, appended to a carboxamido group, as defined 0088 "Arylsulfinyl refers to Rss S(O)—, wherein Rss herein. is an aryl group, as defined herein. 0105 Arylcarboxamido” refers to an aryl group, as 0089) “Arylsulfonyl refers to Rss S(O) , wherein defined herein, appended to a carboxamido group, as defined R is an aryl group, as defined herein. herein. 0090 “Amidyl” refers to Rs.C(O)N(Rs.)— wherein Rs. 0106). “Urea refers to N(Rs.) C(O)N(R)(Rs.) and Rs, are each independently a hydrogen atom, an alkyl wherein Rs. Rs7, and Rs are each independently a hydrogen group, an aryl group, an alkylaryl group, or an arylheterocy atom, an alkyl group, an aryl group, an alkylaryl group, or an clic ring, as defined herein. arylheterocyclic ring, as defined herein, or Rs and Rs, taken 0091) “Ester” refers to RC(O)O— wherein Rs is a together with the nitrogen to which they are attached form a hydrogen atom, an alkyl group, an aryl group, an alkylaryl heterocyclic ring, as defined herein. group, or an arylheterocyclic ring, as defined herein. 0107 “Phosphoryl” refers to - P(R)(R)(R), 0092 “Carbamoyl refers to —O C(O)N(R)(R), wherein Rio is a lone pair of electrons, Sulfur or oxygen, and wherein Rs, and Rs 7 are each independently a hydrogenatom, R7, and R7 are each independently a covalent bond, a hydro an alkyl group, an aryl group, an alkylaryl group or an aryl gen, a lower alkyl, an alkoxy, an alkylamino, a hydroxy or an heterocyclic ring, as defined herein, or Rs, and Rs, taken aryl, as defined herein. together are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein. 0.108 “Silyl refers to —Si(R-7) (Ra)(R-7s), wherein Rz. R7 and Rs are each independently a covalent bond, a lower 0093. “Carbamate” refers to RO-C(O)N (Rs), alkyl, an alkoxy, an aryl or an arylalkoxy, as defined herein. wherein Rs, and Rs 7 are each independently a hydrogenatom, an alkyl group, an aryl group, an alkylaryl group or an aryl 01.09) “Hydrazino” refers to HN N(H)–. US 2009/0226407 A9 Sep. 10, 2009

0110) "Hydralazine compound” refers to a compound Sion (e.g., low-renin hypertension; salt-sensitive hyperten having the formula: Sion; low-renin, salt-sensitive hypertension; primary pulmo nary hypertension; thromboembolic pulmonary hypertension; pregnancy-induced hypertension; renovascu R4 R3 lar hypertension; hypertension-dependent end-stage renal a. b C disease), heart failure (e.g., microvascular cardiac ischemia), RN - N - R2 and left ventricular hypertrophy with disproportionate microvascularization, (i.e., inadequate vascularity) or dias whereina, b and care independently a single or double bond; tolic dysfunction. R and R2 are each independently a hydrogen, an alkyl, an 0113) "Cardiovascular diseases” refers to any cardiovas ester or a heterocyclic ring, wherein alkyl, ester and hetero cular disease, including but not limited to, congestive heart cyclic rind are as defined herein; R and Ra are each indepen failure, hypertension, pulmonary hypertension, myocardial dently alone pair of electrons or a hydrogen, with the proviso and cerebral infarctions, atherosclerosis, atherogenesis, that at least one of R. R. R. and R is not a hydrogen. thrombosis, ischemic heart disease, post-angioplasty resteno Exemplary hydralazine compounds include budralazine, sis, coronary artery diseases, renal failure, stable, unstable cadralazine, dihydralazine, endralazine, hydralazine, and variant (Prinzmetal) angina, cardiac edema, renal insuf pildralazine, todralazine, and the like. ficiency, nephrotic edema, hepatic edema, stroke, transient 0111 "Compound used to treat cardiovascular diseases” ischemic attacks, cerebrovascular accidents, restenosis, con refers to any therapeutic compound, or a pharmaceutically trolling blood pressure in hypertension, platelet adhesion, acceptable salt thereof, used to treat any cardiovascular dis platelet aggregation, Smooth muscle cell proliferation, vascu ease. Suitable compounds include, but are not limited to, lar complications associated with the use of medical devices, angiotensin-converting enzyme (ACE) inhibitors (such as, wounds associated with the use of medical devices, pulmo for example, alacepril, benazepril, captopril, ceronapril, nary thromboembolism, cerebral thromboembolism, throm cilaZapril, delapril, duinapril, enalapril, enalaprilat, fosino bophlebitis, thrombocytopenia, bleeding disorders, and the pril, imidapril, lisinopril, losartan, moveltipril, naphthopidil, like. pentopril, perindopril, quinapril, ramipril, rentipril, spirapril, 0114 “Diseases resulting from oxidative stress' refers to temocapril, trandolapril, urapidil. Zofenopril, and the like): any disease that involves the generation of free radicals or beta-adrenergic blockers (such as, for example, acebutolol, radical compounds, such as, for example, atherogenesis, alprenolol, amosulalol, arotinolol, atenolol, betaxolol, atheromatosis, arteriosclerosis, artherosclerosis, vascular bethanidine, bevantolol, bisoprolol, bopindolol, bucumolol, hypertrophy associated with hypertension, hyperlipoprotein bufetolol, bufuralol, bunitrolol, bupranolol, butafilolol, cara aemia, normal vascular degeneration through aging, parathy Zolol, carteolol, carvedilol, celiprolol, cetamolol, dilevalol, roidal reactive hyperplasia, chronic renal disease, neoplastic epanolol, esmolol, indenolol, labetalol, mepindolol, metipra diseases, inflammatory diseases, neurological and acute nolol, metoprolol, moprolol, nadolol, nadoxolol, nebivolol. bronchopulmonary disease, tumorigenesis, ischemia-reper nifenalol, nipradillol, Oxprenolol, penbutolol, pindolol, prac fusion syndrome, arthritis, sepsis, and the like. tolol, pronethalol, propranolol, Sotalol, sulfinalol, talinolol. tertatolol, tilisolol, timolol, toliprolol. xibenolol, and the 0115 Two broad classes of cardiovascular disorders are like); cholesterol reducers (such as, for example, HMG-CoA more prevalent among blacks than whites and serve as areas reductase inhibitors, including, but not limited to, lovastatin in need of investigative efforts. Hypertension and left ven (MEVACORR), simvastatin (ZOCOR(R), pravastatin (PRA tricular hypertrophy, two related yet independent risk factors VACHOL(R), fluvastatin, cerivastatin (BAYCOLOR), atorvas for coronary heart disease, are significantly more prevalent tatin (LIPITORR), and the like: sequestrants, including, but among blacks than whites. Blacks also have higher rates of not limited to, cholestyramine, colestipol, sialkylaminoalkyl angiographically normal coronary arteries despite a higher derivatives of cross-linked dextran, and the like: inhibitors of prevalence of risk factors for coronary atherosclerosis, and cholesterol absorption, including, but not limited to, beta greater morbidity and mortality from coronary heart disease sitosterol, acyl CoA-cholersterol acyltransferase inhibitors, than whites. These paradoxical observations have led some melinamide, and the like); calcium channel blockers (such as, investigators to postulate that blacks harbor a diathesis of the for example, amlodipine, aranidipine, bamidipine, benid microvasculature that limits perfusion and serves as a stimu ipine, cilnidipine, clentiazem, diltiazen, efonidipine, fanto lus for vascular smooth muscle cell and cardiomyocyte farone, felodipine, isradipine, lacidipine, lercanidipine, hypertrophy, which, in turn, leads to hypertension and left manidipine, mibefradil, nicardipine, nifedipine, nilvadipine, Ventricular hypertrophy, respectively. The underlying basis nisoldipine, nitrendipine, semotiadil. Veraparmil, and the for this vascular diathesis may involve the endothelium, like); angiotensin II receptor antagonists (such as, for which has a limited capacity to generate vasodilator and example, ciclosidomine, eprosartan, furosemide, irbesartan, antiproliferative factors or an increased capacity to produce losartan, Saralasin, Valsartan, and the like); endothelin vasoconstrictor and proliferative factors; the vascular smooth antagonists (such as, for example, bosentan, sulfonamide muscle cell, which manifests increased sensitivity to vaso endothelin antagonists, BQ-123, SQ 28.608, and the like): constrictor and proliferative factors; or both, in these indi renin inhibitors (such as, for example, enalkrein, RO viduals. 42-5892. A 65317, CP80794, ES 1005, ES 8891, SQ34017, 0116) The present inventors have discovered that a major and the like); and mixtures thereof. product of the normal blood vessel that may play a role in the 0112 “Vascular diseases characterized by nitric oxide vascular diathesis of blacks is endothelium-derived nitric insufficiency” include, but are not limited to, cardiovascular oxide (NO). Nitric oxide produced by the endothelial cells diseases; diseases resulting from oxidative stress; hyperten induces vascular smooth muscle cell relaxation, contributing US 2009/0226407 A9 Sep. 10, 2009

importantly to resting vascular tone. In addition, NO inhibits sequent cardiovascular disorders common among African vascular Smooth muscle cell proliferation and induces apop Americans. Possible candidate mechanisms for the oxidant tosis in smooth muscle cells, which leads to the release of stress include enhanced production of reactive oxygen spe basic fibroblast growth factor and vascular endothelial cell cies (ROS), decreased antioxidant defenses, or both. The growth factor, in turn Supporting endothelial cell prolifera inventors make no a priori assumptions about the temporal or tion. This sequence of cellular responses is believed to Sustain causative relationship between oxidant stress and the vascular angiogenesis under hypoxic or ischemic conditions. phenotype of blacks: oxidant stress may both precede the development of the vascular diathesis and promote its pro 0117 The role of nitric oxide in the vascular diathesis of gression once established. Recent data Suggest that enhanced blacks is illustrated by the consequences of nitric oxide insuf ROS production accompanies essential hypertension, athero ficiency in the normal responses of the vasculature to nitric Sclerosis, thrombosis, and diabetes mellitus, and appears in oxide. Nitric oxide insufficiency Suppresses renin release each case, at the very least, to be important in the progression from the juxtaglomerular cells, and induces a Sodium chlo ride/volume sensitive increase in blood pressure. Further of established disease, if not in its actual genesis. more, nitric oxide insufficiency leads to an increased sensi 0123 Endothelium-derived relaxing factor (EDRF), first tivity of vascular Smooth muscle cells to vasoconstrictors, described by Furchgottetal, Nature, 299:373-376 (1980), is Such as angiotensin II and catecholamines, which amplify the an important mediator of vascular function. This endothelial increase in vascular resistance. product activates guanylyl cyclase invascular Smooth muscle 0118 Nitric oxide insufficiency promotes vascular cells and platelets, leading to vasorelaxation and platelet inhi Smooth muscle cell proliferation following vascular injury, bition, respectively (Loscalzo et al. Prog Cardiovasc Dis, and Sustains Smooth muscle cell and cardiomyocyte hyper 38:87-104 (1995)). The chemical nature of EDRF has been trophy in response to catecholamines and angiotensin II. Fur studied using a variety of pharmacological and analytical thermore, inadequate nitric oxide leads to increased produc techniques, and is NO (Ignarro et al. Circ Res, 61:866-879 tion of extracellular matrix with consequent myocardial (1987); Palmer et al, Nature, 327:524-526 (1987)). fibrosis. 0.124 Nitric oxide is synthesized by one of several iso 0119) These many cardiovascular responses that result forms of the NO synthase (NOS) family of enzymes, two of from inadequate NO in the vasculature have clear clinical which are found in the vasculature, endothelial NOS (eNOS) correlates in the black population. The clinical vascular phe and inducible NOS (iNOS). eNOS is synthesized by endot notype of blacks that distinguishes them from whites with helial cells, while iNOS is synthesized by a variety of cell similar cardiovascular disorders is one of salt-sensitive, low types, including vascular Smooth muscle cells, fibroblasts, renin hypertension; left ventricular hypertrophy dispropor and (principally microvascular) endothelial cells (Balligand tionate to afterload and with an inadequate angiogenic etal, Am J Physiol, 268: H1293-1303 (1995)). These enzymes response, and microvascular ischemia in the absence of Sig produce NO as a result of the five-electron oxidation of nificant epicardial coronary artery disease. The net patho L-arginine to L-citrulline; requisite cofactors include cal physiological consequences of these effects are increased cium-calmodulin, O, FAD, FMN, tetrahydrobiopterinthiols, peripheral vascular resistance with accompanying arterial heme, and NADPH. (Moncadaetal, NEnglJMed, 329:2002 hypertension; and an inadequately vascularized, fibrotic 2012 (1993)). increase in left ventricular mass with accompanying diastolic 0.125 The role of NO in the cardiovascular system has dysfunction and microvascular ischemia. become increasingly apparent over the past fifteen years 0120 Given these clinical observations and the role that (Loscalzo et al. Prog Cardiovasc Dis, 38:87-104 (1995)). NO plays in preventing their development, the present inven Nitric oxide contributes importantly to resting tone in con ductance as well as resistance arteries (Ouyyumi et al., J. Clin tors have unexpectedly discovered that the principal cardio Invest, 95:1747-1755 (1995)), and plays a critical role in the vascular disorders common among blacks (such as hyperten maintenance of peripheral vascular resistance and arterial sion, left ventricular hypertrophy, and heart failure) result pressure responses. Inhibition of NOS activity is associated from a specific vascular diathesis that is a direct consequence with enhanced vascular sensitivity to vasoconstrictors. Such of nitric oxide insufficiency. An outline of the pathogenic as norepinephrine and angiotensin II (Conrad et al. Am J consequences of nitric oxide insufficiency that serve as the Physiol, 262:R1137-R1144 (1992)), and this effect appears to basis for these cardiovascular disorders is shown in FIG. 1. be mediated, in part, by increased calcium sensitivity (Banket 0121 Nitric oxide insufficiency states can be a conse al, Hypertension, 24:322-328 (1994)). Nitric oxide release quence of reduced synthesis of nitric oxide, enhanced inacti from the cardiovascular regulatory center in the brain may vation of nitric oxide, or both. Possible candidate mecha also be involved in the central regulation of blood pressure, nisms include alterations in the genes that code for Suggesting a role for neuronal NOS in the regulation of vas endothelial nitric oxide synthase or the inducible microvas cular tone (Cabrera et al. Biochem Biophy's Res Comm, cular and cardiomyocyte nitric oxide synthase leading to 206:77-81 (1995); Mattson et al., Hypertension, 28:297-303 reduced expression of a normal gene product or appropriate (1996)). expression of a less active gene product; reduction in the 0.126 Nitric oxide activates renin gene expression in the enzymatic activity of nitric oxide synthase owing to inad kidney, and is involved in the baroreceptor-mediated regula equate cofactor concentrations; or enhanced inactivation of tion of renin gene expression (Schricker et al., Pflug Arch, nitric oxide by oxidant stress. 428:261-268 (1994)). The dependence of blood pressure on 0122 Data obtained by the inventors in cultured cells, salt intake appears to depend on NO, and NO deficiency states animal models, and human patients suggest that increased are associated with salt-sensitivity (Tolins etal, Kidney Inter oxidant stress is central to the vascular diathesis of and con nat, 46:230-236 (1994)). Selective inhibition of iNOS in Dahl US 2009/0226407 A9 Sep. 10, 2009

R rats has been shown to lead to salt-sensitivity and to the tion, including increased platelet activation, arterial throm development of salt-dependent hypertension similar to Dahl bosis (Freedman et al, J Clin Invest, 97:979-987 (1996): Srats (Rudd etal, Am J Physiol, 277:H732-H739 (1999)). In Freedman et al. Circulation, 98:1481-1486 (1998)), and addition, mice deficient in iNOS (iNOS gene eliminated by reduced production of platelet-derived NO (Kenet etal, Arte targeted disruption) may develop hypertension in response to rio Thromb Vasc Biol, 19(8): 2017-2023 (1999)), which is salt feeding (Rudd et al. Circulation, 98:1A (1998)). important for limiting expansion of a platelet thrombus 0127. Nitric oxide also affects myocardial contractility, (Freedman et al., Circ Res, 84:1416-142 (1999)). and does so both by mediating muscarinic-cholinergic slow 0131 ROS generation accompanies the vascular dysfunc ing of the heart rate and the contractile response to beta tion associated with several models of atherothrombotic and adrenergic stimulation (Balligand et al. Proc Nat'l Acad Sci hypertensive vascular diseases. Hyperhomo-cysteinemic USA, 90:347-351 (1993)). This latter effect appears to be mice (i.e., cystathionine B-synthase knock-out mice) (Eber mediated in vivo through the vagus nerve (Hare et al., J. Clin hardt et al. Circulation, 98:144 (1998)), cellular glutathione Invest, 95:360-366 (1995)). peroxidase-deficient mice (i.e., cellular glutathione peroxi dase knock-out mice), and salt-induced hypertensive rats 0128. In both vascular smooth muscle cells and cardi (i.e., salt-fed Dahl Srats) (Trollietetal, Circulation, 98:1-725 omyocytes, NO inhibits cellular proliferation and limits the (1998)) all manifest increased vascular ROS, and this proliferative response to growth-promoting Substances (Garg increase in ROS is accompanied by reduced NO bioactivity et al., J Clin Invest, 83:1774-1777 (1986)). Left ventricular through oxidative inactivation. Endothelial function and NO hypertrophy tends to occur in adult hearts with inadequate availability can be improved by improving antioxidant status capillary proliferation, and this may account for the with a cysteine precursor (Vita et al., J. Clin Invest, 101: 1408 microvascular ischemia noted in patients with hypertrophy. 1414 (1998)). In addition, alpha-tocopherol leads to platelet Capillary proliferation is generally held to be a rare event in inhibition (Freedman et al. Circulation, 94:2434-2440 normal adult mammalian hearts. However, recent data from a (1996)) as one mechanism of its atherothrombotic benefit hypertensive rat model, in which left ventricular hypertrophy (Stephens et al. Lancet, 347:781-786 (1996)). The present commonly occurs, show that treatment with a low-dose of an inventors have also discovered that salt-loading salt-sensitive angiotensin-converting enzyme inhibitor insufficient to pre individuals (Dahl S rats) leads to an approximate 5-fold vent hypertension and left ventricular hypertrophy can, none increase in plasma F-isoprostanes (8-epi-prostaglandin F), theless, evoke capillary angiogenesis. Compared with and this increase precedes the development of florid hyper untreated controls, treatment with the angiotensin converting tension. These data all support the role of oxidant stress in the enzyme inhibitor increased myocardial capillary prolifera genesis or evolution of vascular dysfunction and disease, and tion (Unger et al. Hypertension, 20:478482 (1992)), and this the importance of antioxidant mechanisms in preventing this effect was believed to be a consequence of inhibiting the pathobiology, particularly with regard to African Americans. degradation and potentiating the action of bradykinin. Brady kinin increases myocardial blood flow by inducing release of 0.132. In support of the mechanisms illustrated above, NO from microvascular endothelial cells, and increased minimum forearm vascular resistance is significantly higher blood flow is a powerful stimulus for capillary proliferation among normotensive blacks than whites (Bassett et al. An J (Mall et al. Bas Res Cardiol, 85:531-540 (1990)). Hypertension, 5:781-786 (1992)), and forearm blood-flow responses to isoproterenol are markedly attenuated in normo 0129 Normal metabolic processes in vascular cells are tensive blacks, suggesting a blunted B-vasodilator response associated with the generation of reactive oxygen intermedi in these individuals (Lang et al., N Engl J Med, 333:155-160 ates that must be neutralized to limit oxidative damage and (1995)). Blacks tend to have greater left ventricular mass than cellular dysfunction. In the setting of common cardiovascular whites for any given level of blood pressure (Koren et al. Am disorders or in the presence of common risk factors for J Hypertension, 6:815-823 (1993); Chaturvedi et al, J Am atherothrombotic disease, reactive oxygen species (ROS) are Coll Cardiol, 24:1499-1505 (1994)). While not quantitated in generated in abundance, and their rate of synthesis and flux any necropsy study, this response is likely to be accompanied typically exceeds the capacity of endogenous antioxidant by inadequate capillary angiogenesis which, in turn, may mechanisms. Hypercholesterolemia, hyperglycemia (Keaney account for the diastolic dysfunction and the microvascular et al. Circulation, 99:189-191 (1999)), cigarette smoking, ischemia observed in blacks. Interestingly, blacks have been hyperhomocysteinemia, hypertension, and frank atheroscle observed to have low levels of urinary kallikrein (Zinneretal, rosis are all accompanied by an increase in plasma and tissue Am J Epidemiol, 104:124-132 (1976); Levy et al. J Clin ROS generation. Superoxide anion, hydrogen peroxide, Invest, 60:129-138 (1977)), the enzyme responsible for the hydroxyl radical, peroxynitrite, and lipid peroxides all generation of bradykinin from high-molecular-weight kini increase in these settings. What remains unknown is whether nogen. Thus, were a similar abnormality in bradykinin and or not the increase in ROS in these disorder is a primary event, bradykinin-mediated NO production to exist in the coronary a secondary consequence of the underlying process, or both. vasculature, attenuated blood flow responses may result that 0130. Endogenous antioxidants important for the neutral would limit capillary angiogenic responses and prevent the ization (i.e., reduction) of ROS can be categorized into two endothelial proliferative effects of locally derived NO. groups: Small-molecule antioxidants and antioxidant 0.133 As discovered and described herein, African Ameri enzymes. The former group comprises molecules such as cans have a unique vascular diathesis that may serve as the GSH, NADPH, alpha-tocopherol, vitamin C, and ubiquinol basis for clinically important cardiovascular syndromes. For 10; while the latter group comprises the Superoxide dismu example, differences in the outcome of left ventricular dys tases, catalase, and glutathione peroxidases. Deficiencies in function may be a consequence of the enhanced (perhaps several of these molecular species have been shown to lead to salt-dependent) increase in oxidant stress coupled with increased steady-state levels of ROS and vascular dysfunc microvascular endothelial dysfunction and an inadequately US 2009/0226407 A9 Sep. 10, 2009

vascularized, hypertrophied left ventricle. This constellation 0.137 Another aspect of the present invention provides of pathophysiological abnormalities may provide the Sub novel transdermal patches comprising atherapeutically effec strate for the important differences in outcome between tive amount of at least one antioxidant and at least one com blacks and whites with left ventricular dysfunction (Dreis et pound that donates, transfers or releases nitric oxide, elevates al, N Engll J Med, 340:609-616 (1999)). In addition, these endogenous levels of endothelium-derived relaxing factor, observations and their clinical consequences Suggest that stimulates endogenous synthesis of nitric oxide or is a Sub blacks with abnormal endothelial function and nitric oxide strate for nitric oxide synthase, and, optionally, at least one insufficiency states would derive direct and, perhaps, dispro nitrosated angiotensin-converting enzyme inhibitor, nitro sated beta-adrenergic blocker, nitrosated calcium channel portionate clinical benefit from enhancing nitric oxide in the blocker, nitrosated endothelin antagonist, nitrosated angio vasculature, either by improving endothelial function, pro tensin II receptor antagonist, nitrosated renin inhibitor, and/or viding exogenous nitric oxide donors, or both. at least one compound used to treat cardiovascular diseases 0134. In view of the above, the present invention provides 0.138 Yet another aspect of the present invention provides methods of treating and/or preventing vascular diseases char Sustained release formulations comprising a therapeutically acterized by nitric oxide (NO) insufficiency by administering effective amount of at least one antioxidant or a pharmaceu a therapeutically effective amount of at least one antioxidant tically acceptable Salt thereof, and at least one nitric oxide or a pharmaceutically acceptable salt thereof, and at least one donor, and, optionally at least one nitrosated compound. compound that donates, transfers or releases nitric oxide, elevates endogenous levels of endothelium-derived relaxing 0.139. In the present invention, the antioxidants include factor, stimulates endogenous synthesis of nitric oxide or is a Small-molecule antioxidants and antioxidant enzymes. Anti Substrate for nitric oxide synthase, and, optionally, at least oxidant refers to and includes any compound that can react one nitrosated angiotensin-converting enzyme inhibitor, nit and quench a free radical. The antioxidant enzymes can be rosated beta-adrenergic blocker, nitrosated calcium channel delivered by gene therapy as a viral vertor and/or a non-viral blocker, nitrosated endothelin antagonist, nitrosated angio vector. Suitable small-molecule antioxidants include, but are tensin II receptor antagonist, nitrosated renin inhibitor, and/or not limited to, hydralazine compounds, glutathione, Vitamin at least one compound used to treat cardiovascular diseases. C. vitamin E. cysteine, N-acetyl-cysteine, 3-carotene, For example, the patient can be administered an antioxidant ubiquinone, ubiquinol-10, tocopherols, coenzyme Q, and the and a nitric oxide donor, or the patient can be administered an like. Suitable antioxidant enzymes include, but are not limited antioxidant, a nitric oxide donor and a nitrosated compound, to, Superoxide dismutase, catalase, glutathione peroxidase, or the patient can be administered an antioxidant, a nitric and the like. Suitable antioxidants are described more fully in oxide donor, a nitrosated compound and a compound used to the literature, such as in Goodman and Gilman, The Pharma treat cardiovascular diseases. cological Basis of Therapeutics (9th Edition), McGraw-Hill, 1995; and the Merck Index on CD-ROM, Twelfth Edition, 0135) Another aspect of the present invention provides Version 12:1, 1996; and on STN Express, file phar and file methods for treating and/or preventing vascular diseases reg. characterized by nitric oxide insufficiency by administering 0140. The preferred antioxidant may be a hydralazine to a patient a therapeutically effective amount of at least one compound that might preferably be administered as a phar nitrosated angiotensin-converting enzyme inhibitor, nitro maceutically acceptable salt; more preferably as hydralazine sated beta-adrenergic blocker, nitrosated calcium channel hydrochloride. Hydralazine hydrochloride (1-hydrazinoph blocker, nitrosated endothelin antagonist, nitrosated angio thalazine monohydrochloride), USP is a white to off-white tensin II receptor antagonist and/or nitrosated renin inhibitor, crystalline powder. It is soluble in water, slightly soluble in and, optionally, at least one antioxidant and/or at least one ethanol and practically insoluble in ether. Hydralazine hydro compound used to treat cardiovascular diseases. chloride is commercially available from, for example, Led 0136. The present invention also provides methods of pre erle Standard Products (Pearl River, N.Y.), and Par Pharma venting and treating Raynaud's syndrome by administering a ceuticals Inc. (Spring Valley, N.Y.). therapeutically effective amount of at least one antioxidant or a pharmaceutically acceptable salt thereof, and at least one 0.141 Compounds contemplated for use in the present compound that donates, transfers or releases nitric oxide, invention (e.g., antioxidants) are used in combination with elevates endogenous levels of endothelium-derived relaxing nitric oxide and compounds that release nitric oxide or oth factor, stimulates endogenous synthesis of nitric oxide or is a erwise directly or indirectly deliver or transfer nitric oxide to Substrate for nitric oxide synthase, and, optionally, at least a site of its activity, Such as on a cell membrane in vivo. one nitrosated angiotensin-converting enzyme inhibitor, nit 0.142 Nitrogen monoxide can exist in three forms: NO rosated calcium channel blocker, nitrosated endothelin (nitroxyl), NOo (nitric oxide) and NO" (nitrosonium). NOo antagonist, nitrosated angiotensin II receptor antagonist and/ is a highly reactive short-lived species that is potentially toxic or nitrosated renin inhibitor. For example, the patient can be to cells. This is critical because the pharmacological efficacy administered an antioxidant and a nitric oxide donor, or the of NO depends upon the form in which it is delivered. In patient can be administered an antioxidant and a nitrosated contrast to the nitric oxide radical (NOo), nitrosonium (NO") compound. The antioxidant, nitric oxide donorand nitrosated does not react with O. or O species, and functionalities compound can be administered separately or as components capable of transferring and/or releasing NO" and NO-are of the same composition. Raynaud's syndrome is a condition also resistant to decomposition in the presence of many redox that causes a loss of blood flow to the fingers, toes, nose and/or metals. Consequently, administration of charged NO equiva ears. The affected area turns white from the lack of circula lents (positive and/or negative) does not result in the genera tion, then blue and cold, and finally numb. The affected area tion of toxic by-products or the elimination of the active NO may also turn red, and may throb, tingle or Swell. moiety. US 2009/0226407 A9 Sep. 10, 2009

0143. The term “nitric oxide' encompasses uncharged 0150 (iii) HN CH(COH)–(CH), C(O)NH nitric oxide (NOo) and charged nitrogen monoxide species, CH(CHSNO) C(O)NH-CH COH: preferably charged nitrogen monoxide species, such as nitrosonium ion (NO") and nitroxyl ion (NO-). The reactive 0151 wherein m is an integer from 2 to 20; R and Rare form of nitric oxide can be provided by gaseous nitric oxide. each independently a hydrogen, an alkyl, a cycloalkoxy, The nitric oxide releasing, delivering or transferring com a halogen, a hydroxy, an hydroxyalkyl, an alkoxyalkyl, pounds, have the structure F-NO, wherein F is a nitric oxide an arylheterocyclic ring, an alkylaryl, a cycloalkylalkyl, releasing, delivering or transferring moiety, include any and a heterocyclicalkyl, an alkoxy, a haloalkoxy, an amino, all such compounds which provide nitric oxide to its intended an alkylamino, a dialkylamino, an arylamino, a diary site of action in a form active for its intended purpose. The lamino, an alkylarylamino, an alkoxyhaloalkyl, a term "NO adducts' encompasses any nitrogen monoxide haloalkoxy, a Sulfonic acid, a Sulfonic ester, an alkylsul releasing, delivering or transferring compounds, including, fonic acid, an arylsulfonic acid, an arylalkoxy, an alky for example, S-nitrosothiols, nitrites, nitrates, S-nitrothiols, lthio, an arylthio, a cycloalkylthio, a cycloalkenyl, a sydnonimines, 2-hydroxy-2-nitrosohydrazines (NONOates), cyano, an aminoalkyl, an aminoaryl, an aryl, an aryla (E)-alkyl-2-((E)-hydroxyimino)-5-nitro-3-hexene amines or lkyl, an alkylaryl, a carboxamido, a alkylcarboxamido, amides, nitroSoamines, furoxans as well as Substrates for the an arylcarboxamido, an amidyl, a carboxyl, a carbam endogenous enzymes which synthesize nitric oxide. The “NO oyl, a carbamate, an alkylcarboxylic acid, an arylcar adducts' can be mono-nitrosylated, poly-nitrosylated, mono boxylic acid, an alkylcarbonyl, an arylcarbonyl, an ester, nitrosated and/or poly-nitrosated or a combination thereof at a carboxylic ester, an alkylcarboxylic ester, an arylcar a variety of naturally susceptible or artificially provided bind boxylic ester, a haloalkoxy, a Sulfonamido, an alkylsul ing sites for biologically active forms of nitrogen monoxide. fonamido, an arylsulfonamido, a Sulfonic ester, aurea, a phosphoryl, a nitro, -T-Q, or (C(R)(R)) T Q, or R. 0144 One group of NO adducts is the S-nitrosothiols, and R taken together with the carbons to which they are which are compounds that include at least one —S NO attached form a carbonyl, a methanthial, a heterocyclic group. These compounds include S-nitroso-polypeptides (the ring, a cycloalkyl group or a bridged cycloalkyl group;Q term “polypeptide' includes proteins and polyamino acids is —NO or - NO; and T is independently a covalent that do not possess an ascertained biological function, and bond, a carbonyl, an oxygen, —S(O) or derivatives thereof); S-nitrosylated amino acids (including —N(R)R , wherein o is an integer from 0 to 2, R is a natural and synthetic amino acids and their stereoisomers and lone pair of electrons, a hydrogen oran alkyl group; R is racemic mixtures and derivatives thereof); S-nitrosylated a hydrogen, an alkyl, an aryl, an alkylcarboxylic acid, an Sugars; S-nitrosylated, modified and unmodified, oligonucle aryl carboxylic acid, an alkylcarboxylic ester, an aryl otides (preferably of at least 5, and more preferably 5-200 carboxylic ester, an alkylcarboxamido, an arylcarboxa nucleotides); straight or branched, saturated or unsaturated, mido, an alkylaryl, an alkylsulfinyl, an alkylsulfonyl, an aliphatic or aromatic, Substituted or unsubstituted S-nitrosy arylsulfinyl, an arylsulfonyl, a Sulfonamido, a carboxa lated hydrocarbons; and S-nitroso heterocyclic compounds. mido, a carboxylic ester, an amino alkyl, an amino aryl, S-nitrosothiols and methods for preparing them are described —CH, CCT-Q)(R)(R), or—(N.O. )M", wherein in U.S. Pat. Nos. 5,380,758 and 5,703,073; WO 97/27749; M is an organic or inorganic cation; with the proviso WO98/19672; and Oae etal, Org. Prep. Proc. Int., 15(3):165 that when R, is —CH, CCT-Q)(R)(R) or —(N.O. 198 (1983), the disclosures of each of which are incorporated )M; then "-T-Q' can be a hydrogen, an alkyl group, an by reference herein in their entirety. alkoxyalkyl group, an aminoalkyl group, a hydroxy 0145 Another embodiment of the present invention is group or an aryl group. S-nitroso amino acids where the nitroso group is linked to a Sulfur group of a Sulfur-containing amino acid or derivative 0152. In cases where R and Rare a heterocyclic ring or thereof. Such compounds include, for example, S-nitroso-N- RandR when taken together with the carbonatoms to which acetylcysteine, S-nitroso-captopril, S-nitroso-N-acetylpeni they are attached are a heterocyclic ring, then R can be a cillamine, S-nitroso-homocysteine, S-nitroso-cysteine, S-ni Substituent on any disubstituted nitrogen contained within the troso-glutathione and S-nitroso-cysteinyl-glycine. radical wherein R, is as defined herein. 0146 Suitable S-nitrosylated proteins include thiol-con 0153. Nitrosothiols can be prepared by various methods of taining proteins (where the NO group is attached to one or synthesis. In general, the thiol precursor is prepared first, then more Sulfur groups on an amino acid oramino acid derivative converted to the S-nitrosothiol derivative by nitrosation of the thereof) from various functional classes including enzymes, thiol group with NaNO, under acidic conditions (pH is about Such as tissue-type plasminogen activator (TPA) and cathep 2.5) which yields the S-nitroso derivative. Acids which can be sin B; transport proteins, such as lipoproteins; heme proteins, used for this purpose include aqueous Sulfuric, acetic and Such as hemoglobin and serum albumin; and biologically hydrochloric acids. The thiol precursor can also be nitrosy protective proteins, such as immunoglobulins, antibodies and lated by reaction with an organic nitrite Such as tert-butyl cytokines. Such nitrosylated proteins are described in WO nitrite, or a nitrosonium salt Such as nitrosonium tetrafluo 93/09806, the disclosure of which is incorporated by refer roborate in an inert solvent. ence herein in its entirety. Examples include polynitrosylated 0154 Another group of NO adducts for use in the present albumin where one or more thiol or other nucleophilic centers invention, where the NO adduct is a compound that donates, in the protein are modified. transfers or releases nitric oxide, include compounds com prising at least one ON O ON N— or ON.—C 0147. Other examples of suitable S-nitrosothiols include: group. The compounds that include at least one ON-O-, 0148 (i) HS(C(R)(R)), SNO; ON N or ON C group are preferably ON O , 0149 (ii) ONS(C(R)(R)), Re; and ON N or ON C-polypeptides (the term “polypeptide' US 2009/0226407 A9 Sep. 10, 2009 includes proteins and polyamino acids that do not possess an nitrosylated analogs (e.g., nitrosated L-arginine, nitrosylated ascertained biological function, and derivatives thereof); L-arginine, nitrosated N-hydroxy-L-arginine, nitrosylated ON O, ON N or ON C-amino acids (including natural N-hydroxy-L-arginine, nitrosated L-homoarginine and and synthetic amino acids and their stereoisomers and race nitrosylated L-homoarginine), precursors of L-arginine and/ mic mixtures); ON O ON N— or ON C-sugars; or physiologically acceptable salts thereof, including, for ON O. , ON N or ON C modified or unmodified example, citrulline, ornithine, glutamine, lysine, polypep oligonucleotides (comprising at least 5 nucleotides, prefer tides comprising at least one of these amino acids, inhibitors ably 5-200 nucleotides); ON O. , ON N- or ON-C of the enzyme arginase (e.g., N-hydroxy-L-arginine and 2CS)- straight or branched, saturated or unsaturated, aliphatic or amino-6-boronohexanoic acid) and the Substrates for nitric aromatic, Substituted or unsubstituted hydrocarbons; and oxide synthase, cytokines, adenosin, bradykinin, calreticulin, ON O—, ON N— or ON C-heterocyclic compounds. bisacodyl, and phenolphthalein. EDRF is a vascular relaxing factor secreted by the endothelium, and has been identified as 0155 Another group of NO adducts for use in the present nitric oxide (NO) or a closely related derivative thereof invention include nitrates that donate, transfer or release nitric (Palmeretal, Nature, 327:524-526 (1987); Ignarro etal, Proc. oxide. Such as compounds comprising at least one ON Natl. Acad. Sci. USA, 84:9265-9269 (1987)). O—, ON N ON S or ON C group. Preferred among these compounds are ON-O-ON-N-, ON.— 0159. In the present invention the compound that donates, S— or ON C polypeptides (the term “polypeptide' transfers or releases nitric oxide as a charged species, or includes proteins and also polyamino acids that do not pos elevates levels of endogenous EDRF or nitric oxide, or is a sess an ascertained biological function, and derivatives substrate for nitric oxide synthase may preferably be isosor thereof); ON O ON N—, ON S or ON.—C bide dinitrate and/or isosorbide mononitrate, more preferably amino acids (including natural and synthetic amino acids and isosorbide dinitrate. Diluted isosorbide dinitrate (1.4.3,6-di their stereoisomers and racemic mixtures); ON-O , anhydro-D-glucitol-2,5-dinitrate), USP is a white to off ON N. , ON S or ON C-sugars; ON O , white powder that has a melting point of 70° C. and has an ON N ON S or ON C modified and unmodi optical rotation of +135° (3 mg/mL, ethanol). It is freely fied oligonucleotides (comprising at least 5 nucleotides, pref soluble in organic solvents such as ethanol, ether and chloro erably 5-200 nucleotides); ON O—, ON. N. , ON.— form, but is sparingly soluble in water. Isosorbide dinitrate is S— or ON-C - Straight or branched, Saturated or commercially available, for example, under the trade names unsaturated, aliphatic or aromatic, Substituted or unsubsti DILATRATE(R) SR (Schwarz Pharma, Milwaukee, Wis.): tuted hydrocarbons; and ON O ON N ON S ISORDILR and ISORDILR TITRADOSER (Wyeth Labo or ON C heterocyclic compounds. Preferred examples ratories Inc., Philadelphia, Pa.); and SORBITRATER (Zen of compounds comprising at least one ON-O-, ON.— eca Pharmaceuticals, Wilmington, Del.). Isosorbide mononi N—, ON S or ON C group include isosorbide trate is commercially available, for example, under the trade dinitrate, mononitrate, clonitrate, erythrityl tetranitrate, man names IMDUR(R) (A. B. Astra, Sweden); MONOKETR) nitol hexanitrate, nitroglycerin, pentaerythritoltetranitrate, (Schwarz Pharma, Milwaukee, Wis.); and ISMOR (Wyeth pentrinitrol and propatylnitrate, most preferred are isosorbide Ayerst company, Philadelphia, Pa.). dinitrate and/or isosorbide mononitrate. 0.160 “Nitrosated compound” refers to any compound 0156 Another group of NO adducts are N-oxo-N-ni that has been Substituted with a nitro group, i.e., —NO troSoamines that donate, transfer or release nitric oxide and group. The nitrosated angiotensin-converting enzyme inhibi are represented by the formula: R'R''N N(O-M'.) NO, tors, nitrosated beta-adrenergic blockers, nitrosated calcium where R' and R are each independently a polypeptide, an channel blockers, nitrosated endothelin antagonists, nitro amino acid, a Sugar, a modified or unmodified oligonucle sated angiotensin II receptor antagonists and nitrosated renin otide, a straight or branched, saturated or unsaturated, ali inhibitors of the present invention include any known angio phatic or aromatic, Substituted or unsubstituted hydrocarbon, tensin-converting enzyme inhibitors, beta-adrenergic block or a heterocyclic group, and M is as defined herein. ers, calcium channel blockers, endothelin antagonists, angio tensin II receptor antagonists and renin inhibitors that have 0157 Another group of NO adducts are thionitrates that been nitrosated through one or more sites such as oxygen donate, transfer or release nitric oxide and are represented by (hydroxyl condensation), Sulfur (sulfhydryl condensation), the formula: R'—(S) NO, where R' is a polypeptide, an and/or nitrogen. The nitrosated compounds of the present amino acid, a Sugar, a modified or unmodified oligonucle invention can be prepared using conventional methods known otide, a straight or branched, saturated or unsaturated, ali to one skilled in the art. For example, known methods for phatic or aromatic, Substituted or unsubstituted hydrocarbon, nitrosating compounds are described in U.S. Pat. Nos. 5,380, or a heterocyclic group. Preferred are those compounds 758 and 5,703,073; WO 97/27749; WO 98/19672; and Oaeet where R' is a polypeptide or hydrocarbon with a pair or pairs al, Org. Prep. Proc. Int., 15(3):165-198 (1983), the disclo of thiols that are sufficiently structurally proximate, i.e., vici sures of each of which are incorporated by reference herein in nal, that the pair of thiols will be reduced to a disulfide. their entirety. WO 98/21193 discloses nitrosated ACE inhibi Compounds which form disulfide species release nitroxyl ion tors and nitrosated beta-adrenergic blockers, the disclosure of (NO ) and uncharged nitric oxide (NOo). which is incorporated by reference herein in its entirety. WO 99/00361 discloses nitrate salts of ACE inhibitors, the disclo 0158. The present invention is also directed to compounds sure of which is incorporated by reference herein in its that stimulate endogenous NO or elevate levels of endog entirety. enous endothelium-derived relaxing factor (EDRF) in vivo or are substrates for the enzyme, nitric oxide synthase. Such 0.161 Suitable angiotensin-converting enzyme inhibitors, compounds include, for example, L-arginine, L-homoargin include, but are not limited to, alacepril, benazepril, captopril, ine, and N-hydroxy-L-arginine, including their nitrosated and ceronapril, cilaZapril, delapril, duinapril, enalapril, enalapri US 2009/0226407 A9 Sep. 10, 2009 lat, fosinopril, imidapril, lisinopril, losartan, moveltipril, 0.167 In the methods of the invention, the antioxidant and naphthopidil, pentopril, perindopril, quinapril, ramipril, rent at least one compound that donates, transfers or releases nitric ipril, spirapril, temocapril, trandolapril, urapidil. Zofenopril, oxide, elevates endogenous levels of endothelium-derived and the like. Suitable angiotensin-converting enzyme inhibi relaxing factor, stimulates endogenous synthesis of nitric tors are described more fully in the literature, such as in oxide or is a Substrate for nitric oxide synthase, and, option Goodman and Gilman, The Pharmacological Basis of Thera ally, at least one nitrosated compound, and/or compound used peutics (9th Edition), McGraw-Hill, 1995; and the Merck to treat cardiovascular diseases can be administered as sepa Index on CD-ROM, Twelfth Edition, Version 12:1, 1996; and rate components or as components of the same composition. on STN Express, file phar and file registry. When the antioxidant and at least one nitric oxide donor are 0162 Suitable beta-adrenergic blockers, include, but are administered as separate components for the treatment of not limited to, acebutolol, alprenolol, amoSulalol, arotinolol. vascular diseases characterized by nitric oxide insufficiency atenolol, betaxolol, bethanidine, bevantolol, bisoprolol, or Raynaud's syndrome, they are preferably administered to bopindolol, bucumolol, bufetolol, bufuralol, bunitrolol. the patient at about the same time. About the same time” bupranolol, butafilolol, carazolol, carteolol, carvedilol, celip means that within about thirty minutes of administering one rolol, cetamolol, dilevalol, epanolol, esmolol, indenolol. compound (e.g., antioxidant or nitric oxide donor) to the labetalol, mepindolol, metipranolol, metoprolol, moprolol. patient, the other compound (e.g., nitric oxide donor or anti nadolol, nadoxolol, nebivolol, nifemalol, nipradillol, Oxpre oxidant) is administered to the patient. About the same time' nolol, penbutolol, pindolol, practolol, pronethalol, propra also includes simultaneous administration of the compounds. nolol, Sotalol, sulfinalol, tallinolol, tertatolol, tilisolol, timolol, 0.168. In addition to the administration of the combination toliprolol. xibenolol, and the like. Suitable beta-adrenergic of the antioxidant and nitric oxide donor for the treatment of blockers are described more fully in the literature, such as in vascular diseases characterized by nitric oxide insufficiency, Goodman and Gilman, The Pharmacological Basis of Thera the patients can receive digitalis Such as digoxin and/or peutics (9th Edition), McGraw-Hill, 1995; and the Merck diuretics and/or at least one nitrosated angiotensin-convert Index on CD-ROM, Twelfth Edition, Version 12:1, 1996; and ing enzyme inhibitor, nitrosated beta-adrenergic blocker, nit on STN Express, file phar and file registry. rosated calcium channel blocker, nitrosated endothelin 0163 Suitable calcium channel blockers, include, but are antagonist, angiotensin II receptor antagonist, nitrosated not limited to, amlodipine, aranidipine, barnidipine, benid renin inhibitor, and/or at least one compound used to treat ipine, cilnidipine, clentiazem, diltiazen, efonidipine, fanto cardiovascular diseases. farone, felodipine, isradipine, lacidipine, lercanidipine, 0169. The digoxin is preferably administered orally to manidipine, mibefradil, nicardipine, nifedipine, nilvadipine, achieve a steady state blood serum concentration of at least nisoldipine, nitrendipine, Semotiadil, Veraparmil, and the about 0.7 nanograms per ml to about 2.0 nanograms per ml. like. Suitable calcium channel blockers are described more The diuretic is administered, preferably orally, to manage fully in the literature, such as in Goodman and Gilman, The edema. Suitable diuretics include, but are not limited to, thi Pharmacological Basis of Therapeutics (9th Edition), azides (such as, for example, chlorothiazide, hydrochlorothi McGraw-Hill, 1995; and the Merck Index on CD-ROM, azide); ethacrynic acid, furosemide, Spironalactone, triam Twelfth Edition, Version 12:1, 1996; and on STN Express, file terene or mixtures thereof. Depending on the diuretic used, phar and file registry. potassium may also be administered to the patient in order to 0164 Suitable endothelin antagonists, include, but are not optimize the fluid balance while avoiding hypokalemic alka limited to, bosentan, Sulfonamide endothelin antagonists, losis. The administration of potassium can be potassium chlo BQ-123, SQ28.608, and the like. Suitable endothelin antago ride or by the daily ingestion of foods with high potassium nists are described more fully in the literature, such as in content Such as, for example, bananas, orange juice, and the Goodman and Gilman, The Pharmacological Basis of Thera like. The method of administration of these compounds is peutics (9th Edition), McGraw-Hill, 1995; and the Merck described in further detail in U.S. Pat. No. 4,868,179, the Index on CD-ROM, Twelfth Edition, Version 12:1, 1996; and disclosure of which is incorporated by reference herein in its on STN Express, file phar and file registry. entirety. 0165 Suitable angiotensin II receptor antagonists, 0170 The compounds and compositions of the invention include, but are not limited to, ciclosidomine, eprosartan, can be administered by any available and effective delivery furosemide, irbesartan, losartan, Saralasin, Valsartan, and the system including, but not limited to, orally, bucally, parenter like. Suitable angiotensin II receptor antagonists are ally, by inhalation spray, topically (including transdermally), described more fully in the literature, such as in Goodman and or rectally in dosage unit formulations containing conven Gilman, The Pharmacological Basis of Therapeutics (9th tional nontoxic pharmaceutically acceptable carriers, adju Edition), McGraw-Hill, 1995; and the Merck Index on CD vants, and vehicles as desired. The preferred methods of ROM, Twelfth Edition, Version 12:1, 1996; and on STN administration are by oral administration or topical applica Express, file phar and file registry. tion (transdermal application). 0166 Suitable renin inhibitors, include, but are not limited 0171 Topical administration can also involve the use of to, enalkrein, RO 42-5892, A 65317, CP80794, ES 1005, ES transdermal administration Such as transdermal patches or 8891, SQ34017, and the like). Suitable renin inhibitors are iontophoresis devices. Dosage forms for topical administra described more fully in the literature, such as in Goodman and tion of the compounds and compositions can include creams, Gilman, The Pharmacological Basis of Therapeutics (9th sprays, lotions, gels, ointments, and the like. In such dosage Edition), McGraw-Hill, 1995; and the Merck Index on CD forms, the compositions of the invention can be mixed to form ROM, Twelfth Edition, Version 12:1, 1996; and on STN white, Smooth, homogeneous, opaque cream or lotion with, Express, file phar and file registry. for example, benzyl alcohol 1% or 2% (wit/wt) as a preserva US 2009/0226407 A9 Sep. 10, 2009 tive, emulsifying wax, glycerin, isopropyl palmitate, lactic infusion techniques. Injectable preparations, for example, acid, purified water and sorbitol solution. In addition, the sterile injectable aqueous or oleaginous Suspensions can be compositions can contain polyethylene glycol 400. They can formulated according to the known art using Suitable dispers be mixed to form ointments with, for example, benzyl alcohol ing agents, wetting agents and/or Suspending agents. The 2% (wt/wt) as preservative, white petrolatum, emulsifying sterile injectable preparation can also be a sterile injectable wax, and tenox II (butylated hydroxyanisole, propyl gallate, Solution or Suspension in a nontoxic parenterally acceptable citric acid, propylene glycol). Woven pads or rolls of bandag diluent or solvent, for example, as a solution in 1,3-butane ing material, e.g., gauze, can be impregnated with the com diol. Among the acceptable vehicles and solvents that can be positions in Solution, lotion, cream, ointment or other Such used are water, Ringer's solution, and isotonic sodium chlo form can also be used for topical application. ride solution. Sterile fixed oils are also conventionally used as 0172 The compositions can also be applied topically a solvent or Suspending medium. using a transdermal system, such as one of an acrylic-based 0177. The compounds and compositions of the invention polymer adhesive with a resinous crosslinking agent impreg will typically be administered in a pharmaceutical composi nated with the composition and laminated to an impermeable tion comprising one or more carriers or excipients. Examples backing. In a particular embodiment, the compositions of the of Suitable carriers include, for example, water, silicone, present invention are administered as a transdermal patch, waxes, petroleum jelly, polyethylene glycols, propylene gly more particularly as a Sustained-release transdermal patch. cols, liposomes, Sugars, salt solutions, alcohol, vegetable oils, The transdermal patches of the present invention can include gelatins, lactose, amylose, magnesium Stearate, talc, Surfac any conventional form Such as, for example, adhesive matrix, tants, silicic acids, viscous paraffins, perfume oils, fatty acid polymeric matrix, reservoir patch, matrix or monolithic-type monoglycerides and diglycerides, petroethral fatty acid laminated structure, and are generally comprised of one or esters, hydroxymethylcelluloses, polyvinyl-pyrrolidones, more backing layers, adhesives, penetration enhancers, an and the like. The pharmaceutical preparations can be steril optional rate controlling membrane and a release liner which ized and if desired, mixed with auxiliary agents, e.g., lubri is removed to expose the adhesives prior to application. Poly cants, preservatives, stabilizers, wetting agents, emulsifiers, meric matrix patches also comprise a polymeric-matrix form salts for influencing osmotic pressure, buffers, colorings, fla ing material. Suitable transdermal patches are described in Vorings and/or aromatic Substances and the like which do not more detail in, for example, U.S. Pat. Nos. 5,262,165, 5.948, deleteriously react with the active compounds. For topical 433, 6,010,715 and 6,071,531, the disclosure of each of application, the compositions can also include one or more which are incorporated herein in their entirety. permeation enhancers including, for example, dimethylsul foxide (DMSO), dimethyl formamide (DMF), N,N-dimethy 0173 Solid dosage forms for oral administration can lacetamide (DMA), decylmethylsulfoxide (ClOMSO), poly include capsules, Sustained-release capsules, tablets, Sus ethylene glycol monolaurate (PEGML), glyceral tained release tablets, chewable tablets, sublingual tablets, monolaurate, lecithin, 1-substituted azacycloheptan-2-ones, effervescent tablets, pills, powders, granules and gels. In Such particularly 1-N-dodecylcyclazacycoheptan-2-ones (avail Solid dosage forms, the active compounds can be admixed able under the trademark AZONE from Nelson Research & with at least one inert diluent, such as Sucrose, lactose or Development Co., Irvine Calif.), alcohols and the like. For starch. Such dosage forms can also comprise, as is normal parenteral application, particularly Suitable vehicles consist practice, additional Substances other than inert diluents, e.g., of Solutions, preferably oily or aqueous solutions, as well as lubricating agents, such as magnesium Stearate. In the case of Suspensions, emulsions, or implants. Aqueous Suspensions capsules, tablets, effervescent tablets, and pills, the dosage can contain Substances which increase the viscosity of the forms can also comprise buffering agents. Soft gelatin cap Suspension and include, for example, Sodium carboxymethyl Sules can be prepared to contain a mixture of the active cellulose, Sorbitol and/or dextran. Optionally, the Suspension compound or composition and vegetable oil. Hard gelatin can contain stabilizers. The compositions, if desired, can also capsules can contain granules of the active compound in contain minor amounts of wetting agents, emulsifying agents combination with a solid, pulverulent carrier, such as lactose, and/or pH buffering agents. saccharose, Sorbitol, mannitol, potato starch, corn starch, amylopectin, or cellulose derivatives of gelatin. Tablets and 0.178 The composition, if desired, can also contain minor pills can be prepared with enteric coatings. amounts of wetting agents, emulsifying agents and/or pH buffering agents. The composition can be a liquid solution, 0174 Liquid dosage forms for oral administration can Suspension, emulsion, tablet, pill, capsule, Sustained release include pharmaceutically acceptable emulsions, Solutions, formulation, or powder. The composition can be formulated Suspensions, syrups, and elixirs containing inert diluents as a Suppository, with traditional binders and carriers such as commonly used in the art, Such as water. Such compositions triglycerides. Oral formulations can include standard carriers can also comprise adjuvants, such as wetting agents, emulsi Such as pharmaceutical grades of mannitol, lactose, starch, fying and Suspending agents, and Sweetening, flavoring, and magnesium Stearate, sodium saccharine, cellulose, magne perfuming agents. sium carbonate, and the like. 0175 Suppositories for rectal administration of the com 0.179 Various delivery systems are known and can be used pounds or compositions can be prepared by mixing the drug to administer the compounds or compositions of the present with a Suitable nonirritating excipient Such as cocoa butter invention, including, for example, encapsulation in lipo and polyethylene glycols which are solidat room temperature Somes, microbubbles, emulsions, microparticles, microcap but liquid at rectal temperature, such that they will melt in the Sules, nanoparticles, and the like. The required dosage can be rectum and release the drug. administered as a single unit or in a Sustained release form. 0176) The term parenteral includes subcutaneous injec 0180. The bioavailability of the compositions can be tions, intravenous, intramuscular, intrasternal injection, or enhanced by micronization of the formulations using conven US 2009/0226407 A9 Sep. 10, 2009 tional techniques such as grinding, milling, spray drying and adjusted by one of ordinary skill in the art, will vary depend the like in the presence of suitable excipients or agents such as ing on the age, health, physical condition, sex, diet, weight, phospholipids or Surfactants. extent of the dysfunction of the recipient, frequency of treat 0181 Sustained release dosage forms of the invention may ment and the nature and scope of the dysfunction or disease, comprise microparticles and/or nanoparticles having a thera medical condition of the patient, the route of administration, peutic agent dispersed therein or may comprise the therapeu pharmacological considerations such as the activity, efficacy, tic agent in pure, preferably crystalline, Solid form. For Sus pharmacokinetic and toxicology profiles of the particular tained release administration, microparticle dosage forms compound used, whether a drug delivery system is used, and comprising pure, preferably crystalline, therapeutic agents whether the compound is administered as part of a drug are preferred. The therapeutic dosage forms of this aspect of combination. the present invention may be of any configuration Suitable for 0186 The compounds and compositions of the present Sustained release. invention can be formulated as pharmaceutically acceptable salts. Pharmaceutically acceptable salts include, for example, 0182 Nanoparticle sustained release therapeutic dosage alkali metal salts and addition salts of free acids or free bases. forms are preferably biodegradable and, optionally, bind to The nature of the salt is not critical, provided that it is phar the vascular Smooth muscle cells and enter those cells, pri maceutically-acceptable. Suitable pharmaceutically-accept marily by endocytosis. The biodegradation of the nanopar able acid addition salts may be prepared from an inorganic ticles occurs over time (e.g., 30 to 120 days; or 10 to 21 days) acid or from an organic acid. Examples of Such inorganic in prelysosomic vesicles and lysosomes. Preferred larger acids include, but are not limited to, hydrochloric, hydrobro microparticle therapeutic dosage forms of the present inven mic, hydroiodic, nitric, carbonic, Sulfuric and phosphoric tion release the therapeutic agents for Subsequent target cell acid and the like. Appropriate organic acids include, but are uptake with only a few of the Smaller microparticles entering not limited to, aliphatic, cycloaliphatic, aromatic, heterocy the cell by phagocytosis. A practitioner in the art will appre clic, carboxylic and Sulfonic classes of organic acids, such as, ciate that the precise mechanism by which a target cell assimi for example, formic, acetic, propionic, Succinic, glycolic, lates and metabolizes a dosage form of the present invention gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, depends on the morphology, physiology and metabolic pro maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthra cesses of those cells. The size of the particle sustained release nilic, mesylic, salicylic, p-hydroxybenzoic, phenylacetic, therapeutic dosage forms is also important with respect to the mandelic, embonic (pamoic), methanesulfonic, ethane mode of cellular assimilation. For example, the Smaller nano sulfonic, benzenesulfonic, pantothenic, toluenesulfonic, particles can flow with the interstitial fluid between cells and 2-hydroxyethaneSuifonic, Sulfanilic, Stearic, algenic, B-hy penetrate the infused tissue. The larger microparticles tend to droxybutyric, cyclohexylaminosulfonic, galactaric and be more easily trapped interstitially in the infused primary galacturonic acid and the like. Suitable pharmaceutically tissue, and thus are useful to deliver anti-proliferative thera acceptable base addition salts include, but are not limited to, peutic agents. metallic salts made from aluminum, calcium, lithium, mag 0183 Particular sustained release dosage forms of the nesium, potassium, Sodium and Zinc or organic salts made present invention comprise biodegradable microparticles or from primary, secondary and tertiary amines, cyclic amines, nanoparticles. More particularly, biodegradable micropar N,N'-dibenzylethylene-diamine, chloroprocaine, choline, ticles or nanoparticles are formed of a polymer containing diethanolamine, ethylenediamine, meglumine (N-methyl matrix that biodegrades by random, nonenzymatic, hydro glucamine) and procaine and the like. All of these salts may be lytic Scissioning to release therapeutic agent, thereby forming prepared by conventional means from the corresponding pores within the particulate structure. compound by reacting, for example, the appropriate acid or base with the compound. 0184 In a particular embodiment, the compositions of the present invention are orally administered as a Sustained 0187. In particular embodiments, the hydralazine hydro release tablet or a Sustained release capsule. For example, the chloride can be administered in an amount of about 30 milli Sustained release formulations can comprise atherapeutically grams per day to about 400 milligrams per day; the isosorbide effective amount of at least one antioxidant or a pharmaceu dinitrate can be administered in an amount of about 5 milli tically acceptable Salt thereof, and at least one nitric oxide grams per day to about 200 milligrams per day; and the donor, or the Sustained release formulations can comprise a isosorbide mononitrate can be administered in an amount of therapeutically effective amount of at least one antioxidant or about 5 milligrams per day to about 120 milligrams per day. a pharmaceutically acceptable salt thereof, and at least one In a more particular embodiment, the hydralazine hydrochlo nitric oxide donor, and, optionally at least one nitrosated ride can be administered in an amount of about 50 milligrams compound. In a particular embodiment the Sustain release per day to about 300 milligrams per day; the isosorbide dini formulation comprises hydralazine hydrochloride and isos trate can be administered in an amount of about 20 milligrams orbide dinitrate and/or isosorbide mononitrate. per day to about 160 milligrams per day; and the isosorbide mononitrate can be administered in an amount of about 15 0185. While individual needs may vary, determination of milligrams per day to about 100 milligrams per day. In an optimal ranges for effective amounts of the compounds and/ even more particular embodiment, the hydralazine hydro or compositions is within the skill of the art and can be chloride can be administered in an amount of about 75 milli determined by standard clinical techniques, including refer grams one to four times per day; the isosorbide dinitrate can ence to Goodman and Gilman, supra: The Physician's Desk be administered in an amount of about 40 milligrams one to Reference, Medical Economics Company, Inc., Oradell, N.J., four time per day; and the isosorbide mononitrate can be 1995; and Drug Facts and Comparisons, Inc., St. Louis, Mo., administered in an amount of about 20 milligrams one to four 1993. Generally, the dosage required to provide an effective times per day. The particular amounts of hydralazine and/or amount of the compounds and compositions, which can be isosorbide dinitrate or isosorbide mononitrate can be admin US 2009/0226407 A9 Sep. 10, 2009

istered as a single dose once a day; or in multiple doses several FIG. 3B, the vasodilator responses to methacholine were not times throughout the day; or as a Sustained-release oral for significantly different in black and white normotensive mulation; or as a transdermal Sustained release patch. patients. However, the response to Sodium nitroprusside 0188 The dose of nitric oxide donor in the composition (FIG. 3A) was significantly lower in the black normotensive will be dependent on the specific nitric oxide donor com patients. This finding shows that there is an impairment in the pound and the mode of administration. For example, when vasodilator response exogenous source of NO in black L-arginine is the orally administered nitric oxide donor, it can patients even prior to the development of hypertension, an be administered in an amount of about 3 grams to about 15 observation that would be consistent with a primary rather grams to provide a plasma level in the range of about 0.2 mM than secondary role in the black vascular diathesis. These to about 30 mM. findings are consistent with recently published results by 0189 The present invention also provides pharmaceutical other investigators (Lang et al., N Engl J Med, 333:155-160 kits comprising one or more containers filled with one or (1995); Cardillo et al. Hypertension 31:1235-1239 (1998)). more of the ingredients of the pharmaceutical compounds and/or compositions. Such kits can also include, for example, Example 3 other compounds and/or compositions (e.g., diuretics, digoxin, nitrosated compounds, compounds used to treat car 0193 Black patients have a preponderance of salt-sensi diovascular diseases and the like), a device(s) for administer tive hypertension, and the data described herein show that NO ing the compounds and/or compositions, and written instruc deficiency is a pathogenic mechanism of salt-sensitive hyper tions in a form prescribed by a governmental agency tension in experimental models (Rudd et al. Am J Physiol, regulating the manufacture, use or sale of pharmaceuticals or 277. H732-H739 (1999)). To explore the relation between biological products, which instructions can also reflects salt-sensitivity and endothelium-derived NO action in black approval by the agency of manufacture, use or sale for human patients prior to the development of hypertension, the blood administration. pressure response to salt-loading and salt-deprivation using an established inpatient protocol (Weinberger et al. Hyper EXAMPLES tension, 8: II-127-II-134 (1986)) in a group of normotensive 0190. The following examples are for purposes of illustra black patients was assessed. Briefly, blood pressure was con tion only, and are not intended to limit the scope of the tinuously monitored non-invasively during a salt load specification or claims. (sodium 458 mEq over 24 hours) and during a period of salt depletion (furosemide treatment and sodium intake 10 mEq Example 1 over 24 hours). Patients were considered to be salt-sensitive if mean blood pressure was at least 10 mm Hg higher during the 0191). As described herein, NO deficiency is a central salt loading period (Weinberger etal, Hypertension, 8: II-127 pathophysiologic mechanism for the black vascular diathesis. II-134 (1986)). As shown in FIGS. 4A-B, there were trends To examine this issue in forearm microVessels, the vasodila for impaired vasodilator responses to both methacholine tor responses to intra-arterial infusions of methacholine, Sodium nitroprusside, and Verapamil were examined using (FIG. 4A) and sodium nitroprusside (FIG. 4B). The vasodi venous occlusion plethysmography in 36 white and black lator responses to Verapamil were equivalent in salt-sensitive hypertensive patients. These patients had no other coronary and salt-resistant patients (data not shown). These data show factors, such as Smoking, diabetes mellitus, or hypercholes impaired NO action in the microvasculature of salt sensitive terolemia, and the two groups were matched in terms of age, individuals prior to the development of hypertension. gender, lipid levels, blood pressure, and anti-hypertensive treatment. Similar to previous reports (Lang et al., N Engl J Example 4 Med, 333:155-160 (1995); Panza etal, NEngl.JMed, 323:22 27 (1990)), the dilator response to methacholine, but not 0194 The effects of hypertension and race on conduit nitroprusside, was significantly reduced in these hypertensive vessel function were examined using a well-established bra patients compared to age-matched normotensive controls chial ultrasound technique (Vita et al. Lanzer & Lipton, Eds. (Sherman et al. Circulation, 98:1-376 (1998)). Regarding Diagnostics of Vascular Diseases. Principles and Technol racial differences, as shown in FIG. 2, vasodilation in ogy. Berlin:Springer-Verlag, pp.249-259 (1996)) in 370 response to methacholine was markedly worse in the black patients (178 black, 192 white). As shown in Table 1, there hypertensive patients compared to white hypertensive were no significant differences in flow-mediated dilation or patients. There were no racial differences in the responses to nitroglycerin-mediated dilation according to race. However, Sodium nitroprusside or Verapamil (data not shown), Suggest hypertension was associated with a highly significant reduc ing that this impairment of NO action in black hypertensives tion in both flow-mediated dilation and nitroglycerin-medi is at the endothelial level and that dysfunction of vascular ated dilation. Further, systolic blood pressure was inversely Smooth muscle does not account for the impaired response. correlated with flow-mediated dilation (r=-0.30, P-0.001) These findings have important implications for the pathogen and nitroglycerin-mediated dilation (r=-0.33, P-0.001). Diastolic pressure correlated to a similar extent with vascular esis of hypertension and myocardial ischemia. function. By multiple linear regression analysis, Vessel size Example 2 and systolic blood pressure were the only independent pre dictors of flow-mediated dilation in this sizable group of 0192 To investigate the issue of whether NO deficiency is patients. Thus, both black and white patients with hyperten a primary or secondary phenomenon in hypertensive black sion demonstrate a significant impairment of NO action in patients, 46 normotensive white and black patients who were conduit arteries (Gokce et al. Circulation, 99(25) 3234-3240 matched for age and gender were compared. As shown in (1999)). US 2009/0226407 A9 Sep. 10, 2009 16

hypertension, myocardial infarction, cerebral infarction, ath TABLE 1. erosclerosis, atherogenesis, thrombosis, ischemic heart dis ease, post-angioplasty restenosis, coronary artery diseases, Conduit Vasonotor Function. By Race renal failure, stable, unstable or variant (Prinzmetal) angina, White Patients Black Patients P cardiac edema, renal insufficiency, nephrotic edema, hepatic edema, stroke, transient ischemic attack, cerebrovascular FMD (%) accident, restenosis, controlling blood pressure in hyperten Normotensive 11.559 13.37.2 NS Sion, platelet adhesion, platelet aggregation, Smooth muscle Hypertensive 9.SS-3* 8.8 6.2* NS cell proliferation, vascular complication associated with the NTG-response (%) use of medical device, wound associated with the use of medical device, pulmonary thromboembolism, cerebral Normotensive 18.1 - 8.1 20.6 - 8.4 NS thromboembolism, thrombophlebitis, thrombocytopenia or Hypertensive 14.6 6.2* 16.47.1* NS bleeding disorder. Data are meant SD. p < 0.001 compared to race matched normotensives. 5. The method of claim 4, wherein the cardiovascular dis ease is congestive heart failure, hypertension, restenosis or 0.195 Each of the above examples demonstrate that NO atherosclerosis. action is impaired in the microvasculature of black hyperten 6. The method of claim 3, wherein the disease resulting sive patients to a greater extent than in white hypertensive from oxidative stress is atherogenesis, atheromatosis, arterio patients. There is a suggestion that this abnormality may Sclerosis, artherosclerosis, vascular hypertrophy associated precede the development of hypertension, particularly in Salt with hypertension, hyperlipoproteinaemia, normal vascular sensitive individuals, consistent with a pathogenic role. In degeneration through aging, parathyroidal reactive hyperpla conduit vessels, a marked impairment of NO action that may sia, chronic renal disease, a neoplastic disease, an inflamma contribute to ischemic heart disease and stroke has been dem tory disease, a neurological and acute bronchopulmonary onstrated. This abnormality appears to be independent of disease, tumorigenesis, ischemia-reperfusion syndrome, race, a finding that is consistent with possibility that the arthritis or sepsis. impairment is a consequence of blood pressure elevation of 7. The method of claim 1, wherein the at least one nitro any cause. Since African Americans have a greater incidence sated angiotensin-converting enzyme inhibitor, nitrosated of hypertension and a preponderance of salt-sensitive hyper beta-adrenergic blocker, nitrosated cholesterol reducer, nit tension, these findings lend further support to the hypothesis rosated calcium channel blocker, nitrosated angiotensin II that nitric oxide insufficiency contributes to the pathogenesis receptor antagonist, nitrosated endothelin antagonist, nitro of ischemic heart disease in this population. sated renin inhibitor, or mixture of two or more thereofare in 0196. The disclosure of each patent, patent application the form of a pharmaceutically acceptable salt. 8. The method of claim 1, wherein the at least one nitro and publication cited herein is hereby incorporated by refer sated angiotensin-converting enzyme inhibitor, nitrosated ence herein in its entirety. beta-adrenergic blocker, nitrosated cholesterol reducer, nit 0197) Although the invention has been set forth in detail, rosated calcium channel blocker, nitrosated angiotensin II one skilled in the art will appreciate that numerous changes receptor antagonist, nitrosated endothelin antagonist, nitro and modifications can be made to the invention without sated renin inhibitor, or mixture of two or more thereof, are departing from the spirit and scope thereof. orally administered as a Solid dose. What is claimed is: 9. The method of claim8, wherein the solid dose is a tablet 1. A method of treating and/or preventing a vascular dis or a capsule. ease characterized by nitric oxide insufficiency in a patient in 10. The method of claim 9, wherein the tablet is a sustained need thereof comprising administering a therapeutically release tablet or the capsule is a Sustained release capsule. effective amount of at least one nitrosated angiotensin-con 11. The method of claim 1, wherein the nitrosated angio Verting enzyme inhibitor, nitrosated beta-adrenergic blocker, tensin-converting enzyme inhibitor is a nitrosated alacepril, a nitrosated cholesterol reducer, nitrosated calcium channel nitrosated benazepril, a nitrosated captopril, a nitrosated blocker, nitrosated angiotensin II receptor antagonist, nitro ceronapril, a nitrosated cilaZapril, a nitrosated delapril, a nit sated endothelin antagonist, nitrosated renin inhibitor, or a rosated duinapril, a nitrosated enalapril, a nitrosated enalap mixture of two or more thereof. rilat, a nitrosated fosinopril, a nitrosated imidapril, a nitro 2. The method of claim 1, further comprising administer sated lisinopril, a nitrosated moveltipril, a nitrosated ing a pharmaceutically acceptable carrier. naphthopidil, a nitrosated pentopril, a nitrosated perindopril, 3. The method of claim 1, wherein the vascular disease a nitrosated quinapril, a nitrosated ramipril, a nitrosated rent characterized by nitric oxide insufficiency is a cardiovascular ipril, a nitrosated spirapril, a nitrosated temocapril, a nitro disease; a disease resulting from oxidative stress; low-renin sated trandolapril, a nitrosated urapidilora nitrosated Zofeno hypertension; salt-sensitive hypertension; low-renin, salt pril. sensitive hypertension; primary pulmonary hypertension; 12. The method of claim 1, wherein the nitrosated beta thromboembolic pulmonary hypertension; pregnancy-in adrenergic blocker is a nitrosated acebutolol, a nitrosated duced hypertension; renovascular hypertension; hyperten alprenolol, a nitrosated amoSulalol, a nitrosated arotinolol, a Sion-dependent end-stage renal disease; heart failure; nitrosated atenolol, a nitrosated betaxolol, a nitrosated microvascular cardiac ischemia; left ventricular hypertrophy bethanidine, a nitrosated bevantolol, a nitrosated bisoprolol, a with disproportionate microvascularization or diastolic dys nitrosated bopindolol, a nitrosated bucumolol, a nitrosated function. bufetolol, a nitrosated bufuralol, a nitrosated bunitrolol, a 4. The method of claim 3, wherein the cardiovascular dis nitrosated bupranolol, a nitrosated butafilolol, a nitrosated ease is congestive heart failure, hypertension, pulmonary carazolol, a nitrosated carteolol, a nitrosated carvedilol, a US 2009/0226407 A9 Sep. 10, 2009

nitrosated celiprolol, a nitrosated cetamolol, a nitrosated dil one antioxidant or a pharmaceutically acceptable salt thereof evalol, a nitrosated epanolol, a nitrosated esmolol, a nitro and/or a therapeutically effective amount of at least one com sated indenolol, a nitrosated labetalol, a nitrosated mepin pound that donates, transfers, or releases nitric oxide, or dolol, a nitrosated metipranolol, a nitrosated metoprolol, a induces the production of endogenous nitric oxide or endot nitrosated moprolol, a nitrosated nadolol, a nitrosated nadox helium-derived relaxing factor or is a substrate for nitric olol, a nitrosated nebivolol, a nitrosated nifenalol, a nitrosated oxide synthase or a pharmaceutically acceptable salt thereof. nipradillol, a nitrosated oXprenolol, a nitrosated penbutolol, a 24. The method of claim 23, wherein the antioxidant is a nitrosated pindolol, a nitrosated practolol, a nitrosated prone Small-molecule antioxidant, or a pharmaceutically accept thalol, a nitrosated propranolol, a nitrosated Sotalol, a nitro able salt thereof, or an antioxidant enzyme. sated Sulfinalol, a nitrosated talinolol, a nitrosated tertatolol, a 25. The method of claim 24, wherein the small-molecule nitrosated tilisolol, a nitrosated timolol, a nitrosated toliprolol antioxidant is a compound of formula (I), a glutathione, a or a nitrosated Xibenolol, Vitamin C, a vitamin E, a cysteine, a N-acetyl-cysteine, a 13. The method of claim 1, wherein the nitrosated choles carotene, an ubiquinone, an ubiquinol-10, a tocopherol, a terol reducer is a nitrosated HMG-CoA reductase inhibitor, a coenzyme Q, or a mixture of two or more thereof; wherein the nitrosated sequestrant or a nitrosated inhibitor of cholesterol compound of formula (I) is: absorption. 14. The method of claim 13, wherein the nitrosated HMG CoA reductase inhibitor is a nitrosated lovastatin, a nitrosated simvastatin, a nitrosated pravastatin, a nitrosated fluvastatin, a nitrosated cerivastatin or a nitrosated atorvastatin. 15. The method of claim 13, wherein the nitrosated seques trant is a nitrosated cholestyramine, a nitrosated colestipol or a nitrosated alkylaminoalkyl derivatives of cross-linked dex whereina, b and c are independently a single or double bond; tran. R and R2 are each independently a hydrogen, an alkyl, an 16. The method of claim 13, wherein the nitrosated inhibi ester or a heterocyclic ring; R and Rare each independently tor of cholesterol absorption is a nitrosated beta-sitosterol, a alone pair of electrons or a hydrogen; with the proviso that at nitrosated acyl CoA-cholersterol acyltransferase inhibitor or least one of R. R. R. and R is not a hydrogen. a nitrosated melinamide. 26. The method of claim 23, wherein the antioxidant 17. The method of claim 1, wherein the nitrosated calcium enzyme is a Superoxide dismutase, a catalase, a glutathione channel blocker is a nitrosated amlodipine, a nitrosated arani peroxidase, or a mixture of two or more thereof. dipine, a nitrosated bamidipine, a nitrosated benidipine, a 27. The method of claim 25, wherein the compound of nitrosated cilnidipine, a nitrosated clentiaZem, a nitrosated formula (I) is budralazine, cadralazine, dihydralazine, diltiazen, a nitrosated efonidipine, a nitrosated fantofarone, a endralazine, hydralazine, pildralazine, todralazine or a phar nitrosated felodipine, a nitrosated isradipine, a nitrosated maceutically acceptable salt thereof. lacidipine, a nitrosated lercanidipine, a nitrosated manid 28. The method of claim 25, wherein the compound of ipine, a nitrosated mibefradil, a nitrosated nicardipine, a nit formula (I) is hydralazine hydrochloride. rosated nifedipine, a nitrosated nilvadipine, a nitrosated 29. The method of claim 23, wherein the at least one nisoldipine, a nitrosated nitrendipine, a nitrosated semotiadil compound that donates, transfers, or releases nitric oxide, or or a nitrosated Veraparmil. induces the production of endogenous nitric oxide or endot 18. The method of claim 1, wherein the nitrosated angio helium-derived relaxing factor or is a substrate for nitric tensin II receptor antagonist is a nitrosated ciclosidomine, a oxide synthase is an S-nitrosothiol. nitrosated eprosartan, a nitrosated furosemide, a nitrosated 30. The method of claim 29, wherein the S-nitrosothiol is irbesartan, a nitrosated losartan, a nitrosated Saralasin or a S-nitroso-N-acetylcysteine, S-nitroso-captopril, S-nitroso nitrosated Valsartan. N-acetylpenicillamine, S-nitroso-homocysteine, S-nitroso 19. The method of claim 1, wherein the nitrosated endot cysteine, S-nitroso-glutathione or S-nitroso-cysteinyl-gly helin antagonist is a nitrosated bosentan, a nitrosated Sulfona cine. mide endothelin antagonists, a nitrosated BQ-123 or a nitro 31. The method of claim 29, wherein the S-nitrosothiol is: sated SQ28.608. 20. The method of claim 1, wherein the nitrosated renin (i) HS(C(R)(R)), SNO; inhibitor is a nitrosated enalkrein, a nitrosated RO42-5892, a (ii) ONS(C(R)(R)), nesR.; and nitrosated A 65317, a nitrosated CP 80794, a nitrosated ES 1005, a nitrosated ES 8891 or a nitrosated SQ34017. (iii) HN CH(COH)–(CH), C(O)NH 21. The method of claim 1, further comprising administer CH(CHSNO) C(O)NH CH, COH: ing to the patientatherapeutically effective amount of at least wherein m is an integer from 2 to 20; R and Rare each one compound used to treat a cardiovascular disease or a independently a hydrogen, an alkyl, a cycloalkoxy, a pharmaceutically acceptable salt thereof. halogen, a hydroxy, anhydroxyalkyl, an alkoxyalkyl, an 22. The method of claim 21, wherein the compound used to arylheterocyclic ring, an alkylaryl, a cycloalkylalkyl, a treat the cardiovascular disease is an angiotensin-converting heterocyclicalkyl, an alkoxy, a haloalkoxy, an amino, an enzyme inhibitor, a beta-adrenergic blocker, a cholesterol alkylamino, a dialkylamino, an arylamino, a diary reducer, a calcium channel blocker, an angiotensin II receptor lamino, an alkylarylamino, an alkoxyhaloalkyl, a antagonist, an endothelin antagonist, a renin inhibitor, or a haloalkoxy, a Sulfonic acid, a Sulfonic ester; an alkylsul mixture of two or more thereof. fonic acid, an arylsulfonic acid, an arylalkoxy, an alky 23. The method of claim 1, further comprising administer lthio, an arylthio, a cycloalkylthio, a cycloalkenyl, a ing to the patientatherapeutically effective amount of at least cyano, an aminoalkyl, an anoaryl, an aryl, an arylalkyl, US 2009/0226407 A9 Sep. 10, 2009 18

an alkylaryl, a carboxamido, a alkylcarboxamido, an independently a polypeptide, an amino acid, a Sugar, an arylcarboxamido, an amidyl, a carboxyl, a carbamoyl, a oligonucleotide, a straight or branched, Saturated or carbamate, an alkylcarboxylic acid, an arylcarboxylic unsaturated, aliphatic or aromatic, Substituted or unsub acid, an alkylcarbonyl, an arylcarbonyl, an ester, a car stituted hydrocarbon, or a heterocyclic group, and M+is boxylic ester, an alkylcarboxylic ester, an arylcarboxylic an organic or inorganic cation. ester, a haloalkoxy, a Sulfonamido, an alkylsulfonamido, an arylsulfonamido, a Sulfonic ester, a urea, a phospho 35. The method of claim 34, wherein the compound com ryl, a nitro, -T-Q, or —(C(R)(R))-T-Q, or R and Rf prising at least one ON O—, ON N— or ON C group taken together with the carbons to which they are is an ON O-polypeptide, an ON N-polypeptide, an attached form a carbonyl, a methanthial, a heterocyclic ON C-polypeptide, an ON O-amino acid, an ON N ring, a cycloalkyl group or a bridged cycloalkyl group;Q amino acid, an ON-C-amino acid, an ON-O-Sugar, an is —NO or —NO; and T is independently a covalent ON N-sugar, an ON C-sugar, an ON O-oligonucle bond, a carbonyl, an oxygen, —S(O) or otide, an ON N-oligonucleotide, an ON C-oligonucle —N(R)R , wherein o is an integer from 0 to 2, R is a otide, a straight or branched, Saturated or unsaturated, Substi lone pair of electrons, a hydrogen oran alkyl group; R is tuted or unsubstituted, aliphatic or aromatic ON O a hydrogen, an alkyl, an aryl, an alkylcarboxylic acid, an hydrocarbon, a straight or branched, saturated or unsaturated, aryl carboxylic acid, an alkylcarboxylic ester, an aryl substituted or unsubstituted, aliphatic or aromatic ON N carboxylic ester, an alkylcarboxamido, an arylcarboxa hydrocarbon, a straight or branched, saturated or unsaturated, mido, an alkylaryl, an alkylsulfinyl, an alkylsulfonyl, an substituted or unsubstituted, aliphatic or aromatic ON C arylsulfinyl, an arylsulfonyl, a Sulfonamido, a carboxa hydrocarbon, an ON-O-heterocyclic compound, an mido, a carboxylic ester, an amino alkyl, an amino aryl, ON N-heterocyclic compound or a ON C-heterocyclic —CH2—C(TQ)(R)(R), or —(NO )oM", wherein compound. M" is an organic or inorganic cation; with the proviso 36. The method of claim 34, wherein compound compris that when R is —CH2—C(T-Q)(R)(R) or —(NO )'M', then"-T-Q' can be a hydrogen, an alkyl group, an ing at least one ON O—, ON N - ON S or alkoxyalkyl group, an aminoalkyl group, a hydroxy ON C group is an ON O-polypeptide, an ON N group or an aryl group. polypeptide, an ON S-polypeptide, an ON-C-polypep 33. The method of claim 23, wherein the at least one tide, an ON O-amino acid, ON N-amino acid, ON.— compound that donates, transfers, or releases nitric oxide, or S-amino acid, an ON-C-amino acid, an ON-O-sugar, an induces the production of endogenous nitric oxide or endot ON-N-sugar, ON S-Sugar, an ON C-Sugar, an helium-derived relaxing factor, or is a substrate for nitric ON-O-oligonucleotide, an ON-N-oligonucleotide, an oxide synthase is L-arginine, L-homoarginine, N-hydroxy-L- ON S-oligonucleotide, an ON-C-oligonucleotide, a arginine, nitrosated L-arginine, nitrosylated L-arginine, nit straight or branched, Saturated or unsaturated, aliphatic or rosated N-hydroxy-L-arginine, nitrosylated N-hydroxy-L- aromatic, substituted or unsubstituted ON O-hydrocar arginine, citrulline, omithine, glutamine, lysine, polypeptides bon, a straight or branched, Saturated or unsaturated, aliphatic comprising at least one of these amino acids oran inhibitor of or aromatic, substituted or unsubstituted ON N-hydrocar the enzyme arginase. bon, a straight or branched, Saturated or unsaturated, aliphatic 34. The method of claim 23, wherein the at least one or aromatic, substituted or unsubstituted ON S-hydrocar compound that donates, transfers, or releases nitric oxide, or bon, a straight or branched, Saturated or unsaturated, aliphatic induces the production of endogenous nitric oxide or endot or aromatic, substituted or unsubstituted ON C-hydrocar helium-derived relaxing factor, or is a substrate for nitric bon, an ON O-heterocyclic compound, an ON N-het oxide synthase is: erocyclic compound, an ON S-heterocyclic compound or (i) a compound that comprises at least one ON-O-, an ON-C-heterocyclic compound. ON N or ON C group; 37. The method of claim 36, wherein compound compris ing at least one ON O—, ON N - ON S or (ii) a compound that comprises at least one ON-O-, ON-C - group is isosorbide mononitrate and/or isosor ON N. , ON S or ON C group; bide dinitrate. (iii) a N-oxo-N-nitrosoamine having the formula: R'R''N N(O-M') NO, wherein R' and Rare each