US 2004O1 O1523A1. (19) United States (12) Patent Application Publication (10) Pub. No.: US 2004/0101523 A1 Reitz et al. (43) Pub. Date: May 27, 2004 (54) RENAL-SELECTIVE PRODRUGS FOR Publication Classification CONTROL OF RENAL SMPATHETIC NERVE ACTIVITY IN THE TREATMENT OF (51) Int. Cl. ............................. A61K 38/43; C12N 9/99 HYPERTENSION (52) U.S. Cl. ........................................... 424/94.1; 435/184 (75) Inventors: David B. Reitz, Chesterfield, MO (US); John P. Koepke, St. Louis, MO (US); (57) ABSTRACT Edward H. Blaine, University City, MO (US); Joseph R. Schuh, St. Louis, MO (US); Robert E. Manning, St. Renal-Selective prodrugs are described which are preferen Louis, MO (US); Glenn J. Smits, tially converted in the kidney to compounds capable of Ellisville, MO (US) inhibiting Synthesis of catecholamine-type neurotransmit ters involved in renal Sympathetic nerve activity. The pro drugs described herein are derived from inhibitor com Correspondence Address: pounds capable of inhibiting one or more of the enzymes J. Timothy Keane involved in catecholamine Synthesis, Such compounds being PHARMACIA CORPORATION of Pfizer Inc., classifiable as tyrosine hydroxylase inhibitors, or as dopa Corporate Patent Department decarboxylase inhibitors, or as dopamine-B-hydroxylase P.O. BOX 1027 inhibitors. These inhibitor compounds are linked to a chemi Chesterfield, MO 63006 (US) cal moiety, Such as a glutamic acid derivative, by a cleavable bond which is recognized Selectively by enzymes located predominantly in the kidney. The liberated inhibitor com (73) Assignee: G.D. Searle & Co., Chicago, IL pound is then available in the kidney to inhibit one or more of the enzymes involved in catecholamine Synthesis. Inhi (21) Appl. No.: 10/689,919 bition of renal catecholamine Synthesis can Suppress height ened renal nerve activity associated with Sodium-retention (22) Filed: Oct. 20, 2003 related disorderS Such as hypertension. Conjugates of par Related U.S. Application Data ticular interest are glutamyl derivatives of dopamine-B- hydroxylase inhibitors, of which N-acetyl-y-glutamyl (63) Continuation of application No. 10/151,211, filed on fusaric acid hydrazide (shown below) is preferred. May 20, 2002, now abandoned, which is a continu ation of application No. 09/678,015, filed on Oct. 2, 2000, now abandoned, which is a continuation of N application No. 09/444,888, filed on Nov. 22, 1999, O O CO2H now abandoned, which is a continuation of applica 2n tion No. 08/639,493, filed on Apr. 29, 1996, now N C-N-N-C-CH-CH-CH abandoned, which is a continuation of application No. 08/280,170, filed on Jul. 25, 1994, now abandoned, H. H. -n which is a continuation-in-part of application No. =o PCT/US90/04168, filed on Jul 25, 1990, which is a CH continuation-in-part of application No. 07/386,527, filed on Jul. 27, 1989, now abandoned. Patent Application Publication May 27, 2004 Sheet 1 of 16 US 2004/0101523 A1 Acute In Vivo Effects of Vehicle and Example #3 Conjugate 100 8 O 6 O 4. O -o- vehicle -O- Example #3 2 O O 1S 30 4S 60 Time (minutes) Figure 1 Patent Application Publication May 27, 2004 Sheet 2 of 16 US 2004/0101523 A1 Acute In Vivo Effects of Example #3 Conjugate s-O- Vehicle -O- Example #3 Conjugate O 5 30 4S 60 Tine (minutes) Patent Application Publication May 27, 2004 Sheet 3 of 16 US 2004/0101523 A1 Chronic Infusion of Example #464 Conjugate 2OO Or Vehicle -O- Example #464 Conjugate 190 18O 170 160 150 Time (days) Figure 3 Patent Application Publication May 27, 2004 Sheet 4 of 16 US 2004/0101523 A1 Formation of Fusaric Acid From Example #859 Conjugate by Rat Kidney Homogenate 34.5 2 O O 20 30 40 50 Tine (hours) Patent Application Publication May 27, 2004 Sheet 5 of 16 US 2004/0101523 A1 Enzymatic Formation of Fusaric Acid From Example #859 Conjugate 20 15 O S O 20 40 60 80 OO 120 1 40 Time (hours) Patent Application Publication May 27, 2004 Sheet 6 of 16 US 2004/0101523 A1 Effect of Fusaric Acid and Example #859 Conjugate on Dopamine-6-Hydroxylase Activity In Vitro 0.6 as e N 0.5 2 s - 0. s S. S 03 O2 s s 0.1 -O Fusaric Acid 0.0 -O- Example #859 Conjugate .0 ... 1 10 100 Concentration (M) Figure 6 Patent Application Publication May 27, 2004 Sheet 7 of 16 US 2004/0101523 A1 Dopamine-?3-Hydroxylase Inhibition by Exampie #859 Conjugate and Related Compounds 1 OO Example #859 Conjugate Example #858 Conjugate QP Fusaric Acid Hydrazide us Fusaic Acid Ne 6 O 40 20 Test Compound (20 M) Figure 7 Patent Application Publication May 27, 2004 Sheet 8 of 16 US 2004/0101523 A1 Acute In Vivo Effects of Fusaric Acid or Example #859 Conjugate on Mean Arterial Pressure 18O 160 140 -O- Fusaic Acid -o- Example #859 Conjugate 120 OO 8 O O S 30 4S 60 Tine (minutes) Patent Application Publication May 27, 2004 Sheet 9 of 16 US 2004/0101523 A1 Acute In Vivo Effects of Fusaric Acid and Example #859 Conjugate on Renal Blood Flow TO Fusaric Acid 5 mO Example #859 Conjugate 4. O S 3O 4S 60 Tine (minutes) Patent Application Publication May 27, 2004 Sheet 10 of 16 US 2004/0101523 A1 Chronic In Vivo Effects of Saline, Fusaric Acid and Example #859 Conjugate 200 180 s Vehicle On Fusaric Acid -O-Example #859 Conjugate 160 140 20 100 O 2 3 4 S 6 Time (days) Figure 10 Patent Application Publication May 27, 2004 Sheet 11 of 16 US 2004/0101523 A1 Chronic Infusion of Example #863 Conjugate Or Vehicle 8 O -O- Example #863 Conjugate 170 160 150 140 O 2 3 4 Time (days) Figure 11 Patent Application Publication May 27, 2004 Sheet 12 of 16 US 2004/0101523 A1 Heart Norepinephrine Levels Following 5 Day Infusion of Vehicle, Fusaric Acid, and Example #859 Conjugate 3OOO 2OOO 1 OOO Vehicle Fusaric Acid Example #859 Conjugate Patent Application Publication May 27, 2004 Sheet 13 of 16 US 2004/0101523 A1 Kidney Norepinephrine Levels Following S Day Infusion of Vehicle, Fusaric Acid, and Example #859 Conjugate 1 OOO 68 20 O Vehicle Fusaric Acid ExampleConjugate #859 Figure 13 Patent Application Publication May 27, 2004 Sheet 14 of 16 US 2004/0101523 A1 Mean Arterial Pressure Response to Example #859 Conjugate after I.V. Injection in Dogs 200 Vehicle a 7 mg/kg 150 D 60 mg/kg 125 2 OO % 75 2 50 2% 2 O 2 3 Time (hours) FIGURE 14 Patent Application Publication May 27, 2004 Sheet 15 of 16 US 2004/0101523 A1 Renal Blood Flow Response to Example #859 Conjugate after I.V. Injection in Dogs 200 Vehicle 175 a 7 mg/kg B 20 mg/kg 150 D 60 mg/kg 125 OO 75 50 25 0 1. |2 3 Time (hours) FIGURE 15 Patent Application Publication May 27, 2004 Sheet 16 of 16 US 2004/0101523 A1 Effects of Ex. 859 in DOCA Hypertensive Micropigs 180 -OH vehicleEXAMPLE 200 #859 m/day 5 mg/kg 5 n=3 160 150 140 30 20 110 100 1. 2 3. 4 Time (days) FIGURE 16 US 2004/0101523 A1 May 27, 2004 RENAL-SELECTIVE PRODRUGS FOR CONTROL neys may be an effective therapeutic treatment for chronic OF RENAL SMPATHETIC NERVE ACTIVITY IN Sodium-retaining disorders, Such as hypertension, conges THE TREATMENT OF HYPERTENSION tive heart failure, cirrhosis, and nephrosis. RELATED APPLICATION 0006. One approach to reduce sympathetic nervous sys tem effects on renal function is to inhibit the synthesis of one 0001. This application is a continuation-in-part of U.S. or more compounds involved as intermediates in the "cat Application Ser. No. PCT/US90/04168 filed 25 Jul. 1990, echolamine cascade', that is, the pathway involved in Syn which is a continuation-in-part of U.S. application Ser. No. thesis of the neurotransmitter norepinephrine. Stepwise, 07/386,527 filed 27 Jul 1989. these catecholamines are Synthesized in the following man ner: (1) tyrosine is converted to dopa by the enzyme tyrosine FIELD OF THE INVENTION hydroxylase; (2) dopa is converted to dopamine by the 0002 This invention is in the field of cardiovascular enzyme dopa decarboxylase; and (3) dopamine is converted therapeutics and relates to a class of compounds useful in to norepinephrine by the enzyme dopamine-B-hydroxylase. control of hypertension. Of particular interest is a class of Inhibition of dopamine-B-hydroxylase activity, in particular, compounds which prevent or control hypertension by Selec would increase the renal vasodilatory, diuretic and natri tive action on the renal Sympathetic nervous System. uretic effects due to dopamine. Inhibition of the action of any of these enzymes would decrease the renal vasocon Strictive, antidiuretic and antinatriuretic effects of norepi BACKGROUND OF THE INVENTION nephrine. Therapeutically, these effects oppose chronic 0.003 Hypertension has been linked to increased sympa Sodium retention. thetic nervous System activity Stimulated through any of four mechanisms, namely (1) by increased vascular resistance, 0007 Many compounds are known to inhibit the action (2) by increased cardiac rate, Stroke Volume and output, (3) of the catecholamine-cascade-converting enzymes. For by vascular muscle defects or (4) by Sodium retention and example, the compound C.-methyltyrosine inhibits the action renin release J. P. Koepke et al, The Kidney in Hyperten of the enzyme tyrosine hydroxylase. The compound C.-me Sion, B. M. Brenner and J. H. Laragh (Editors), Vol. 1, p. 53 thyldopa inhibits the action of the enzyme dopa-decarboxy (1987). As to this fourth mechanism in particular, stimula lase, and the compound fuSaric acid inhibits the action of tion of the renal Sympathetic nervous System can affect renal dopamine-B-hydroxylase.