USO08088935B2 (12) United States Patent (10) Patent No.: US 8,088,935 B2 Pears0n et al. (45) Date of Patent: Jan. 3, 2012 (54) COMPOUNDS AND METHODS FOR THE FOREIGN PATENT DOCUMENTS TREATMENT OF ASTHMA DE 2326227 2, 1974 ES 3983.19 4, 1975 Inventors: James Pearson, Cambridge, MA (US); FR 2196795 3, 1974 (75) FR 2196796 3, 1974 John J. Talley, Somerville, MA (US); FR 2210398 7, 1974 Mark G. Currie, Sterling, MA (US) GB 1297264 11, 1972 GB 1374981 11, 1974 (73) Assignee: Ironwood Pharmaceuticals, Inc., WO O193867 12/2001 Cambridge, MA (US) WO O3O806O7 10, 2003 OTHER PUBLICATIONS (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 Wang, 2002, Role of iNOS and eNOS in modulating proximal tubule U.S.C. 154(b) by 1229 days. transport and acid-base balance. Pizza et 1998. Nitric Oxide synthase inhition reduces muscle inflam (21) Appl. No.: 10/587,054 mation. Patrick Vallance 1998. Student BM.J. (22) PCT Fled: Dec. 23, 2004 * cited by examiner (86) PCT NO.: PCT/US2004/043O82 Primary Examiner — Rita Desai S371 (c)(1), (74) Attorney, Agent, or Firm — Heslin Rothenberg Farley (2), (4) Date: Nov. 22, 2006 & Mesiti P.C. (87) PCT Pub. No.: WO2OOS/O63732 (57) ABSTRACT PCT Pub. Date: Jul. 14, 2005 Compounds and methods for the treatment of asthma are disclosed. The methods involve mast cell stabilization (65) Prior Publication Data together with selective inhibition of iNOS. The compounds are combinations of a mast cell inhibiting moiety and an US 2007/O249644 A1 Oct. 25, 2007 inhibitor of iNOS. An example is: Related U.S. Application Data (60) Provisional application No. 60/531,957, filed on Dec. HOOC O O COOH. 23, 2003. (51) Int. C. CO7D 3L/22 (2006.01) A6 IK3I/353 (2006.01) O O ~~ O (52) U.S. Cl. ......................... 549/401; 549/402:514/456 O (58) Field of Classification Search .................. 549/401, 549/402: 514/456 CH O See application file for complete search history. )=NH NH2 (56) References Cited HN U.S. PATENT DOCUMENTS 3,790,580 A 2, 1974 Johnson et al. 2003/O1995.29 A1 10/2003 Garvey et al. 16 Claims, No Drawings US 8,088,935 B2 1. 2 COMPOUNDS AND METHODS FOR THE (1) A neuronal NOS(NOS-1 or nNOS) was originally isolated TREATMENT OF ASTHMA and cloned from nerve tissue in which it is a constitutive enzyme. nNOS produces NO in response to various physi CROSS REFERENCE TO RELATED ological stimuli, such as the activation of membrane recep APPLICATIONS tors, according to a mechanism dependent on calcium and on calmodulin, nNOS-derived NO serves as a neurotransmitter. This application is a 371 filing of PCT International Appli cation No. PCT/US04/43082 filed Dec. 23, 2004, published (2) An inducible NOS(NOS-2 or iNOS) can be induced in in English as WO 2005/063732 on Jul. 14, 2005. PCT/US04/ response to immunological stimuli Such as, for example, 43082 claimed the priority of U.S. Provisional Application 10 cytokines or bacterial antigens in various cells Such as, for 60/531,957 filed Dec. 23, 2033. The disclosures of both are example macrophages, epithelial cells, hepatocytes, glial incorporated herein by reference in their entirety. cells, and other cell types. The activity of this isoform is not regulated by calcium. Once induced, it produces large FIELD OF THE INVENTION 15 amounts of NO over prolonged periods. (3) An endothelial NOS(NOS-3 or eNOS) is constitutive and The invention relates to compounds and methods for the calcium/calmodulin-dependent. It was originally identified in treatment of asthma. The methods involve mast cell stabili vascular endothelial cells, in which it generates NO in zation together with selective inhibition of iNOS. The com response to physiological stimuli such as the activation of pounds are combinations of a mast cell inhibiting moiety and membrane receptors. an inhibitor of iNOS. Nitric oxide produced by eNOS and nNOS plays a critical BACKGROUND OF THE INVENTION role in cellular signaling and acts to control numerous physi ologic functions including vasodilation and bronchodilation Asthma is a chronic airway inflammatory disorder charac 25 in the lung. In the asthmatic lung, eNOS and nNOS are terized by bronchial hyper-reactivity and bronchospasm, downregulated, and thus contribute to edema and bronchoc among other abnormalities. Lungs of asthmatic patients have onstriction. Contemplating the problem of inadequate eNOS increased numbers of inflammatory cells in bronchioalveolar and nNOS activity, in an approach which is the opposite of fluid and in lung tissues. These inflammatory cells include that taken in the present invention, Garvey et al. (US pub eosinophils, basophils, neutrophils, macrophages, and lym 30 lished application 2003/0199529) have attached stimulators phocytes. In asthmatic lungs, the epithelium, including cili of endogenous NO production to mast cell inhibitors. ated columnar epithelial cells, is damaged. IgE-antigen-mast The NO produced in large amounts by the inducible iso cell interactions represent the early molecular and cellular form iNOS is involved in pathological phenomena associated events that cause inflammatory conditions of asthma. 35 with acute and chronic inflammatory processes in a large Mast Cell Stabilizing Agents provide one approach to the variety of tissues and organs. NO is highly reactive and, prophylaxis and/or treatment of asthma. The prototype drug, together with Superoxide, forms peroxynitrite which dam disodium cromoglycate was synthesized in 1965 and was ages tissues. In asthma this results in epithelial cell extrusion, approved in the United States in 1973 as a prophylactic, 40 sloughing, and cessation of cilia function. An excessive pro nonbronchodilating anti-inflammatory drug for the therapy of duction of NO by induction of iNOS thus plays a part in allergic disorder. Cromolyn is an odorless, white, hygro degenerative pathologies with inflammatory components, scopic crystalline powder that is freely soluble in water up to Such as asthma. 5%. Animal and human studies show it to be excreted In conditions in which an overproduction of NO is delete unchanged in bile and urine. When inhaled into the pulmo 45 rious, it would be desirable to reduce the production of NO by nary tree, as for treatment of asthma, only about 8% of a dose administering substances capable of inhibiting iNOS. How is deposited in the lung and absorbed. Peak plasma levels ever, given the important physiological roles played by the occur within 15 minutes, the biologic half-life is 46-99 min constitutive isoforms, selective inhibition of iNOS is utes. Oral administration in humans results in approximately required. 1% being systemically absorbed. Cromolyn toxicity studies 50 show an impressively low order of acute toxicity, and adverse effects tend to be minimal and reversible. Cromolyn has a SUMMARY OF THE INVENTION unique, purely prophylactic action with no intrinsic bron chodilator or antihistaminic activity. Nedocromil was intro 55 In a composition aspect, the invention relates to agents for duced Subsequent to cromolyn. It is the other standard mast treating a pulmonary disorder represented by the structure: cell stabilizer used in the treatment of asthma. Its chemical properties and therapeutic characteristics are similar. Nitric oxide (NO) is a diffusible radical involved in many physiological and pathological processes. It is synthesized in 60 G)--G) Vivo by oxidation of L-arginine. The synthesis is catalyzed by a family of enzymes known as nitric oxide synthases or NO wherein synthases (NOSs), which are referenced in the international A is a mast-cell stabilizer; enzyme nomenclature system under the number 65 E.C.1.14.13.39. Three NOS isoforms, two of which are con L is a covalent linkage; stitutive and one inducible, are known: B is an iNOS inhibitor. US 8,088,935 B2 3 4 Examples of Such agents are compounds of formula I or II G)--G) In a second method aspect, the invention relates to a method for treating a pulmonary disorder comprising co administering a mast-cell stabilizer and an iNOS inhibitor in the form of a salt, in which one of the mast-cell stabilizer and the iNOS inhibitor is a cation or dication, and the other is an anion ordianion. DETAILED DESCRIPTION OF THE INVENTION 15 Agents for treating a pulmonary disorder according to the invention are represented by the structure: II 2O G)--G) In one embodiment, L is chosen from —CONH , CONH- and —(C=O)OCH(R)O(C=O) and the com pound is represented by a structure chosen from: wherein R" and Rare chosen from hydroxy, C.C.C.C.C.s, and C. straight and branched alkoxy, -G-O(C=O)R. R. NHR, OR7 and OX", wherein X" is a pharmaceu tically acceptable cation; 35 losso —O(C=O)R’ is the deshydrogen residue of a carboxylic acid, the parent of which, RCOOH, is an inhibitor of . Grro ro inducible nitric oxide synthase (iNOS); —(C=O)R’ is the deshydroxy residue of a carboxylic acid, the parent of which, RCOOH, is an inhibitor of iNOS; 45 G) Grr. O R is - O R' U, wherein U is chosen from hydrogen, O O G)—o (1,2-dithiolan-3-yl) and phenyl, and R' is a divalent C to r and Coalkane or Oxaalkane residue. O O —NHR is the deshydrogen residue of an amine, the parent of 50 which, RNH is an inhibitor of iNOS; in which A' is a mast-cell stabilizer having a carboxylic acid —OR7 is the deshydrogen residue of an alcohol, the parent of substituent; A is a mast-cell stabilizer having an amine Sub which, ROH, is an inhibitor of iNOS: stituent; A is a mast-cell stabilizer having an alcohol Sub G is a linking moiety cleavable under physiologic conditions.