USOO9623 OOOB2

(12) United States Patent (10) Patent No.: US 9.623,000 B2 Kindler et al. (45) Date of Patent: Apr. 18, 2017

(54) COMPOSITIONS AND METHODS FOR (2013.01); A61L 3 1/16 (2013.01); A61 L TREATING INFLAMMATORY DSORDERS 2300/222 (2013.01); A61 L 2300/41 (2013.01) (58) Field of Classification Search (75) Inventors: Seth Kindler, Tel Aviv (IL); Ascher None Shmulewitz, Tel Aviv (IL) See application file for complete search history. (73) Assignee: DEKEL PHARMACEUTICALS LTD, Bnei-Brak (IL) (56) References Cited (*) Notice: Subject to any disclaimer, the term of this U.S. PATENT DOCUMENTS patent is extended or adjusted under 35 2,554,736 A 5/1951 Haefliger et al. U.S.C. 154(b) by 160 days. 3.420,851 A 1/1969 Bloom et al. 3,438,981 A 4/1969 Stach et al. (21) Appl. No.: 13/056,872 3,505.321 A 4, 1970 Schutz et al. 3,534,041 A 10/1970 Van Der Burg et al. (22) PCT Fed: Jul. 29, 2009 3,934,013 A 1/1976 Poulsen (Continued) (86) PCT No.: PCT/IL2009/OOO739 FOREIGN PATENT DOCUMENTS S 371 (c)(1), (2), (4) Date: Apr. 28, 2011 EP O550006 A2 7, 1993 EP 1452179 A1 9, 2004 (87) PCT Pub. No.: WO2O1 O/O13240 (Continued) PCT Pub. Date: Feb. 4, 2010 OTHER PUBLICATIONS (65) Prior Publication Data International Preliminary Report on Patentability, WIPO (Feb. 1. US 2011 FO195096 A1 Aug. 11, 2011 2011), 6 pages.* Related U.S. Application Data (Continued) (60) Provisional application No. 61/085,008, filed on Jul. Primary Examiner — Brian-Yong Kwon 31, 2008, provisional application No. 61/098,802, Assistant Examiner — Kyung Sook Chang filed on Sep. 22, 2008. (74) Attorney, Agent, or Firm — Vorys, Sater, Seymour & (51) Int. C. Pease LLP A6 IK 3L/20 (2006.01) A 6LX 3/573 (2006.01) (57) ABSTRACT A6 IK 45/06 (2006.01) Compositions which include a corticosteroid in combination A6IL 27/54 (2006.01) with an additional compound active in treatment of an A6IL 3L/6 (2006.01) inflammatory disorder are provided. (52) U.S. C. CPC ...... A61K 31/20 (2013.01); A61K 3 1/573 (2013.01); A61K 45/06 (2013.01); A61L 27/54 7 Claims, 3 Drawing Sheets

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(56) References Cited Kuehl et al., (1957) The identification of N-2 (hydroxyethyl)- palmitamide as a naturally occurring anti-inflammatory agent. JAm U.S. PATENT DOCUMENTS Chem Soc 79: pp. 5577-5578. O. Abdel-Salam, et al.; "Studies on the anti-inflammatory effect of 4,013,639 A 3, 1977 Kitamura et al. floxetine in the rat' Pharmacological Research vol. 49; 2004; pp. 4,105,785 A 8/1978 Mauvernay et al. 119-131. 4,929,637 A 5, 1990 Baldwin et al. 5,006,560 A 4, 1991 Kreutner et al. H. Adalsteinsson, et al.; “Generation and Evaluation of Putative 5,059,613 A 10/1991 Pierce, Jr. Neuroregenerative Drugs. Part 2—Screening Virtual Libraries of 5,242.937 A 9/1993 Pierce, Jr. Novel Polyketides which Possess the Binding Domain of 5,248,678 A 9, 1993 Costa et al. Rapamycin' Bioorganic & Medicinal Chemistry vol. 8; 2000; pp. 5,276,025 A 1/1994 Baldwin et al. 625-635. 5,464,831 A 11/1995 Dean et al. 5,473,067 A 12/1995 Dean et al. A. Bella, et al., “FK1706 Enhances the Recovery of Erectile 5,506,224 A 4, 1996 della Valle et al. Function Following Bilateral Cavernous Nerve Crush Injury in the 5,508,311 A 4, 1996 Yu et al. Rat”; J Sex Med vol. 4; 2007: pp. 341-347. 5,596,106 A 1/1997 Cullinan et al. A. Bella, et al.; “Emerging Neuromodulatory molecules for the 5,646,142 A 7, 1997 Dantanarayana et al. treatment of neurugenic erectile dysfunction caused by cavernous 5,747,524. A 5, 1998 Cullinan et al. nerve injury'; Asian Journal of Andrology vol. 10 Chapter 1; 2008; 5,792,799 A 8, 1998 Sherman-Gold Shanghai Institute of Materia Medica, Chinese Academy of Sci 5,846,979 A 12/1998 Hamilton et al. 6,017,919 A 1/2000 Inaba et al. ences; pp. 54-59. 6,054,452 A 4/2000 Hamilton et al. A. Brown: “Novel Cannabinoid Receptors'; British Journal of 6,077,839 A 6, 2000 WoldeMussie et al. Pharmacology vol. 152; published Online Oct. 2007: pp. 567-575. 6,331,537 B1 12/2001 Hamilton et al. B. Gold; “FK506 and the Role of Immunophilins in Nerve Regen 6,368,814 B1 4/2002 Ghoshal et al. eration”; Molecular Neurobiology vol. 15; 1997; pp. 285-306. 6.432,984 B1 8, 2002 Barth et al. B. Gold: “Neuroimmunophilin ligands—evaluation of their thera 6,472.423 B1 10/2002 Ross et al. peutic potential for the treatment of neurological disorders'; Expert 6,486,151 B2 11/2002 Hamilton et al. 6,509,367 B1 1/2003 Martin et al. Opinion on Investigational Drugs vol. 9; 2000; pp. 2331-2342. 6,569.470 B2 5, 2003 Williams et al. B. Gold, eet al., “A Nonimmunosuppressant FKBP-12 Ligand 6,645,985 B2 11/2003 Barth et al. Increases Nerve Regeneration”: Experimental Neurology vol. 147. 6,656.972 B2 12/2003 Calignano et al. Article No. EN976630; 1997; pp. 269-278. 6,720,348 B2 4/2004 Mylari X. Guo, et al., “Neuroimmunophilin ligands exert neuroregenera 6,821,985 B2 11/2004 Chenard et al. tion and neoroprotection in midbrain dopaminergic neurons'; Euro 6,897.206 B2 * 5/2005 Sackeyfio et al...... 514,171 7,030,250 B2 4/2006 Losada et al. pean Journal of Neuroscience, vol. 13; 2001; pp. 1683-1693. 7,109,353 B2 9/2006 Gurjar et al. A. Heiman, et al., “New Steroidal anti-inflammatory antedrugs'; 7,153,883 B2 12/2006 Hamilton et al. Steroids, Elsevier Science Inc., vol. 62; 1997; pp. 491–499. 7,169,942 B2 1/2007 Moore, II et al. K. Jonsson, et al., “Effects of homologues and analogues of 7,335,371 B2 2/2008 Sackeyfio et al. palmitoylethanolamide upon the inactivation of the endocan 7,348,338 B2 3/2008 Arnold et al. nabinoid anandamide'; British Journal of Pharmacology, vol. 133; 7,544,714 B2 6, 2009 Burstein et al. 2001.0034362 A1 10, 2001 Ross et al. Nature Publishing Group; 2001; pp. 1263-1275. 2002/0173550 A1 11/2002 Calignano et al. D. Long; et al.: “Incorporation of Q Into the Cell Constituents of 2002fO198250 A1 12/2002 Steiner et al. Pseudomonas and E coli Communicaions to the Editor'; vol. 79,

2003/0049220 A1* 3/2003 Bailey ...... A61K 8/34 Oct. 1957; pp. 5577-5578. 424.70.1 J. Kuzmiski; “Topiramate Inhibits the Initiation of Plateau Poten 2003/0186961 A1 10, 2003 Hamilton et al. tials in CA1 Neurons by Depressings R-type Calcium Channels'; 2004/0106975 A1 6/2004 Solovay et al. Epilepsia vol. 46, Chapter 4: Blackwell Publishing, Inc.; Interna 2004/0224876 A1 11/2004 Jost-Price et al. tional League Against Epilepsy; 2005; pp. 481-489. 2005, OO32747 A1* 2, 2005 Bartolini ...... CO7D 209.28 D. Lambert, et al., “Analogues and homologues of 514.80 N-palmitoylethanolamide a putative endogenous CB2 cannabinoid 2005.0054730 A1 3/2005 Fu et al. as potential ligands for the cannabinoid”; Biochemica et Biophysica 2007/0110685 A1 5/2007 Auspitz et al. Acta vol. 1440; 1999: pp. 266-274. 20070203209 A1* 8, 2007 Bartolini et al...... 514,367 D. Lambert, et al.; "The Palmitoylethanolamide Family A New 2009/0118503 A1* 5/2009 Sprott ...... CO7D 209.18 Class of Anti-Inflammatory Agents”; Current Medicinal Chemistry 544, 143 vol. 9; Bentham science Publishers Ltd; 2002: pp. 663-674. 2009/0123504 A1* 5/2009 Feldkamp ... A61K90014 M. Namazi; "Cannabinoids loratadine and allopurinol as novel 424/402 aditions to the antipsoriatic ammunition; European Academy of Dermatology and Venereology; JEADV vol. 19, 2005; pp. 319-322. FOREIGN PATENT DOCUMENTS “Hydrocotarnine” XP-002651747; vol. 4809; p. 855. G. Re, et al.; “Palmitoylethanolamide endocannabinoids and related EP 1645270 A2 4/2006 cannabimimetic compounds in protection agains tissue inflamma WO 9412466 A1 6, 1994 tion and pain Potential use in companion animals'. The Veterinary WO 952O393 A1 8, 1995 Journal vol. 173; 2007: pp. 21-30. WO WOO2O8977O A2 * 11, 2002 E. Roe, et al., “Fatty Acid Amides': Notes vol. 74; Jul. 1952: pp. WO 2005099.720 A2 10, 2005 3442-3443. WO 2007O67.036 A2 6, 2007 WO 2007071313 A2 6, 2007 S. Luan; "Introducing Immuniphilins from Organ Transplantaion to WO 2009 133574 A1 11, 2009 plant Biology': Plant Physiology, vol. 134; Apr. 2004; pp. 1241 1243. D. Sabatini, et al.; “Neural Roles of Immunophilins and Their OTHER PUBLICATIONS Ligands': Molecular Neuobiology vol. 15; 1997; pp. 223-239. H. Sauer, et al.: "Systemic treatment with GPI 1046 improves WO2002089770A2 (claim-english machine translation) (2002).* spatial memory and reserves cholinergic neuron atrophy in the WO2002089770A2 (description—english machine translation) medial septal nucleus of aged mice'; Brain Research vol. 842; 1999; (2002).* pp. 109-118. US 9,623,000 B2 Page 3

(56) References Cited OTHER PUBLICATIONS C. Sich, et al.; "Solution structure of a neotrophic ligand bound to FKBP12 and its effects on protein dynamics': European Journal of Biochemistry vol. 267; Feb. 2000; pp. 5342-5354. J. Steiner, et al., “Neurotrophic immuniphilin ligands stimulate structural and functional reeovery in neurodegenerative animal models”; Proceedings of the National Academy of Sciences USA vol. 94; Neurobiology; Mar. 1997: pp. 2019-2024. J. Steiner, et al., “Neurotrophic actions of nonimmunosuppressive analogues of immunosupressive drugs FK506 rapamycin and cyclosporin A'; Nature Medicine vol. 3, No. 4; Nature Publishing Group; Apr. 1997; pp. 421-428. F. Sun, et al: “Design and Structure-Based Study of New Potential FKBP12 Inhibitors”; Biophysical Journal vol. 85; Nov. 2003; pp. 3194-32O1. G. Sun, et al.; "Carbamazepine and Topiramate Modulation of Transient and Persistent Sodium Currents Studied in HEK293 Cells Expressings the Na 1.3a-Subunit”: Epilepsia vol. 48; Blackwell Publishing, Inc., International League Against Epilepsy; 2007, pp. 774-782. T. Rhen, et al., “Antiinflammatory Action of Glucocorticoids New Mechanism for Old Drugs'. The New England Journal of Medicine vol. 353; Massachusetts Medical Society 2005; pp. 1711-1723. * cited by examiner

US 9,623,000 B2 1. 2 COMPOSITIONS AND METHODS FOR Metal (Lambert DM, DiPaolo FG, Sonveaux P. Kanyonyo TREATING INFLAMMATORY DSORDERS M, Govaerts SJ, Hermans E, Bueb J, Delzenne N M, Tschirhart E. J. Biochim Biophys Acta. 1999; 1440:266-74): CROSS REFERENCE TO RELATED Brown AJ (BrJ Pharmacol. 2007: 152(5):567-75); and U.S. APPLICATIONS 5 Pat. No. 5,506,224 and United States patent application 2005/0054730. None of the above references discloses or This application is a 35 U.S.C. S371 National Phase Entry Suggests use of Such compounds in combination with cor Application from PCT/IL2009/000739, filed Jul. 29, 2009, ticosteroids and the use thereof in treating inflammatory and designating the United States, which claims the benefit diseases or disorders. of U.S. Provisional Patent Application No. 61/085,008, filed 10 Tri-cyclic antidepressants, described interalia in U.S. Pat. on Jul. 31, 2008 and the benefit of U.S. Provisional Patent Nos. 2,554,736, 3,438,981, 3,420,851, 3,534,041, 3,505, Application No. 61/098,802, filed on Sep. 22, 2008, which 321, 4,013,639, 4,105,785, 6,368,814, have been used for are incorporated herein in their entireties. years as anti-depression agents. Topical 5% doxepin hydro chloride (ZonalonTM) cream is also used for treatment of FIELD AND BACKGROUND OF THE 15 moderate pruritus in adult patients with atopic dermatitis or INVENTION lichen simplex chronicus. U.S. Pat. No. 7,335,371, assigned to CombinatoRX, discloses compositions formulated for The present invention relates to compositions suitable for topical administration, comprising as the Sole active ingre treatment of inflammatory disorders and to methods utilizing dients a tricyclic antidepressant and a corticosteroid, and same in treatment of inflammatory disorders such as cuta their use in treating inflammatory disorders. This patent neous inflammatory diseases. neither discloses nor Suggests in any way the ability of Inflammation occurs when tissues are injured by viruses, compositions of the present invention to utilize a low bacteria, trauma, chemicals, heat, cold or any other harmful potency steroid in therapeutic situations wherein a mid stimulus. Chemicals including bradykinin, histamine, sero potency or high-potency steroid would otherwise be tonin and others are released, attracting tissue macrophages 25 required; or to utilize a mid-potency steroid wherein a and white blood cells to localize in an area to engulf and high-potency Steroid would otherwise be required. destroy foreign Substances. During this process, chemical Sulfonamide-class carbonic anhydrase inhibitors such as mediators such as TNFC. are released, giving rise to inflam Sulfamate-substituted monosaccharides, of which topira mation. Inflammatory disorders are those in which the mate is an example, have been long used as anti-convulsive inflammation is Sustained or chronic. One example of an 30 compounds. Topical formulations containing Such com inflammatory disorder is osteoarthritis. pounds are proposed for use in treatment of glaucoma and in Local inflammatory diseases include, for example, the assessment of corneal function, as described in U.S. Pat. asthma, oral mucosal, gastrointestinal inflammation, ocular, Nos. 5,059,613 and 5,242.937, and for eczema, as described nasal and aural inflammation and other steroid responsive in WO2007/067036. None of the above references discloses inflammatory disorders and conditions. Cutaneous inflam 35 or Suggests use of Such compounds in combination with matory diseases include for example, psoriasis, atopic der corticosteroids and their use in treating cutaneous inflam matitis, Scleroderma and other steroid responsive cutaneous matory diseases and disorders. inflammatory disorders conditions such as uremic pruritus Monoamine oxidase inhibitors (MAOI) and used as anti and skin conditions associated with exposure to radiation depressive treatments and are described inter alia in U.S. and chemotherapy as well as exposure to environmental 40 Pat. Nos. 5,508,311, 5,792,799, 6,472,423, and 6,569,470. radiation and irritants. None of the above references discloses or Suggests use of Topical steroids have been used in the treatment of Such compounds in combination with corticosteroids and inflammatory disorders, as described in U.S. Pat. No. 3,934, their use in treating cutaneous inflammatory diseases or 013 to Poulsen and references cited therein. Typically, disorders. moderate to severe cases of psoriasis require use of a 45 Cannabinoids are described inter alia in U.S. Pat. Nos. mid-potency steroid such as mometasone furoate (EloconTM) 5,747,524, 5,596,106, 6,017,919, 6,432,984, 6,509,367, or even a high-potency steroid Such as halobetasol (Ultra 6,645,985, and 7,169,942. Use of such compounds for vate"M). Topical application of glucocorticoids may suppress treating psoriasis has been suggested (Namazi M R. J. Eur the body's own production of cortisol by the adrenal glands, Acad Dermatol Venereol. 2005 May: 19(3):319-22). None however, especially in the case of relatively high potent 50 of the above references discloses or Suggests use of Such products. Accordingly, compositions for treating inflamma compounds in combination with corticosteroids and the use tory disorders without utilizing mid-potency or high-po thereof in treating cutaneous inflammatory diseases or dis tency steroids or with use of substantially reduced steroid orders. concentrations would be very useful. While reducing the present invention to practice, the Palmitoylethanolamine (PEA), a member of the class of 55 present inventors have uncovered that combined use of compounds known as N-acylethanolamines (NAEs), was corticosteroids and PEA, as well as other compounds pro discovered in 1957 by Kuehl et al (The Identification of duces unexpected results in treatment of inflammatory dis N-(2-hydrooxytheyl)-palmitamide as a naturally occurring orders and that PEA has the potential to augment the anti-inflammatory agent, JAm Chem Soc 1957, 79:5577). therapeutic properties of steroids including increasing PEA, or N-(2-hydroxyethyl)hexadecanamide (also known 60 therapeutic activity as well as reducing Steroid associated as palmitoylethanolamide or palmidrol), is a naturally occur adverse effects. ring C16:0 fatty acid derivative wherein the carboxylate function is amidated by the primary amine of ethanolamine. SUMMARY OF THE INVENTION N-acylethanolamine compounds are known to have anti inflammatory and anti-nociceptive effects, as described in 65 According to one aspect of the present invention there is Lambert et al (Lambert D M. Vandevoorde S. Jonsson KO. provided a composition-of-matter comprising an N-acyle Fowler C.J. Curr Med Chem. 2002; 9:663-74); Lambert D thanolamine compound and a corticosteroid. US 9,623,000 B2 3 4 According to further features in preferred embodiments of from the group consisting of amitriptyline, maprotiline, the invention described below, the N-acylethanolamine clomipramine, desipramine, imipramine, trimipramine, nor compound is N-palmitoylethanolamine. triptyline, and protriptyline According to yet another aspect of the present invention According to still further features in the described pre there is provided a composition-of-matter comprising a ferred embodiments the corticosteroid is a low potency GABA agonist and a corticosteroid. corticosteroid. According to still further features in the described pre According to still further features in the described pre ferred embodiments the GABA agonist is selected from the ferred embodiments the composition-of-matter of claim 1 group consisting of topiramate, muscimol, progabide, rilu and a pharmaceutically acceptable carrier. Zole, baclofen, gabapentin, vigabatrin, valproic acid, According to still further features in the described pre 10 tiagabine, lamotrigine, pregabalin, phenyloin, and carbam ferred embodiments the pharmaceutically acceptable carrier aZepine. is Suitable for topical administration. According to yet another aspect of the present invention According to still further features in the described pre there is provided a composition-of-matter comprising a monoamine oxidase inhibitor (MAOI), and a corticosteroid. ferred embodiments the pharmaceutically acceptable carrier 15 is Suitable for mucosal administration. According to still further features in the described pre ferred embodiments the monoamine oxidase inhibitor is According to still further features in the described pre selected from the group consisting of moclobemide, clorgi ferred embodiments the pharmaceutically acceptable carrier line, iproclozide, iproniazid, isocarboxazid, minaprine, nia is suitable for oral administration. lamide, pargyline, phenelzine, rasagiline, selegiline, According to still further features in the described pre toloxatone, tranylcypromine, furazolidone, and procarba ferred embodiments the N-acylethanolamine compound is Z10. N-palmitoylethanolamine. According to still another aspect of the present invention According to still further features in the described pre there is provided a composition-of-matter comprising a ferred embodiments the corticosteroid is hydrocortisone. cannabinoid receptor agonist and a corticosteroid. 25 According to still further features in the described pre According to another aspect of the present invention there ferred embodiments the cannabinoid receptor agonist is is provided a medical device comprising the composition selected from the group consisting of cannabidiol (CBD), of-matter described herein. cannabidivarol (CBDV), cannabinol (CBN), cannabigerol According to still further features in the described pre (CBG), cannabivarol (CBV), cannabicyclol (CBL), tetrahy ferred embodiments the composition-of-matter forms a coat drocannabinol (THC), tetrahydrocannabinol-C4. (THC-C4), ing of the medical device. 30 tetrahydrocannabivarin, 11-Hydroxy-A-tetrahydrocannabi According to still another aspect of the present invention nol, (11-OH-THC), and 11-nor-9-Carboxy-A9-tetrahydro there is provided a method for treating an inflammatory cannabinol. disorder comprising administering to a Subject in need According to still another aspect of the present invention thereof a composition comprising an N-acylethanolamine there is provided a composition-of-matter comprising an 35 inhibitor of an immunophilin and a low-potency corticos compound and a corticosteroid, thereby treating the inflam teroid. matory disorder. According to still further features in the described pre According to still further features in the described pre ferred embodiments the inhibitor of an immunophilin is ferred embodiments the N-acylethanolamine compound is selected from the group consisting of FK 1706, GPI 1046, N-palmitoylethanolamine. 40 GPI 1485, GM-284, (3R)-4-(p-Toluenesulfonyl)-1,4-thiaz According to still further features in the described pre ane-3-carboxylic acid-L-Leucine ethyl ester and (3R)-4-(p- ferred embodiments the corticosteroid is a low-potency Toluenesulfonyl)-1,4-(thiazane-3-carboxylic acid-L-phenyl corticosteroid. alanine benzyl ester. According to still further features in the described pre According to yet another aspect of the present invention ferred embodiments the corticosteroid is hydrocortisone. 45 there is provided a pharmaceutical composition comprising According to still further features in the described pre any of the composition-of-matters described hereinabove ferred embodiments the inflammatory disorder is caused by and a pharmaceutically acceptable carrier selected for topi a cutaneous inflammatory disease or disorder and the admin cal delivery. istering is effected via topical delivery. The present invention successfully addresses the short According to still further features in the described pre 50 comings of the presently known configurations by providing ferred embodiments the cutaneous inflammatory disease is a composition for treating inflammatory disorder which selected from the group consisting of psoriasis, atopic der employs Substantially reduced doses of a corticosteroid. matitis and Scleroderma. Unless otherwise defined, all technical and scientific According to still further features in the described pre terms used herein have the same meaning as commonly ferred embodiments the inflammatory disorder is caused by 55 understood by one of ordinary skill in the art to which this asthma and the administering is effected via inhalation. invention belongs. Although methods and materials similar According to still further features in the described pre or equivalent to those described herein can be used in the ferred embodiments the inflammatory disorder is caused by practice or testing of the present invention, Suitable methods gastrointestinal inflammation. and materials are described below. In case of conflict, the According to still further features in the described pre 60 patent specification, including definitions, will control. In ferred embodiments the inflammatory disorder is caused by addition, the materials, methods, and examples are illustra ocular inflammation. tive only and not intended to be limiting. According to still another aspect of the present invention there is provided a composition-of-matter comprising a BRIEF DESCRIPTION OF THE DRAWINGS corticosteroid and a tricyclic antidepressant. 65 According to still further features in the described pre The invention is herein described, by way of example ferred embodiments the tricyclic antidepressant is selected only, with reference to the accompanying drawings. With US 9,623,000 B2 5 6 specific reference now to the drawings in detail, it is stressed teroid and an additional compound selected for complement that the particulars shown are by way of example and for ing the anti-inflammatory effect of the corticosteroid. purposes of illustrative discussion of the preferred embodi According to a presently preferred embodiment of the ments of the present invention only, and are presented in the present invention, the composition of the present invention cause of providing what is believed to be the most useful and includes a corticosteroid and an N-acylethanolamine com readily understood description of the principles and concep pound such as palmitoylethanolamide (PEA). tual aspects of the invention. In this regard, no attempt is N-acylethanolamine compounds are well known in the made to show structural details of the invention in more art, and are described, interalia, in Lambert et al (Lambert detail than is necessary for a fundamental understanding of DM, Vandevoorde S, Jonsson KO, Fowler C. J. Curr Med the invention, the description taken with the drawings mak 10 Chem. 2002: 9:663-74) and in U.S. Pat. No. 5,506,224 and ing apparent to those skilled in the art how the several forms United States patent application 2005/0054730, which are of the invention may be embodied in practice. incorporated by reference herein. In the drawings: Preferably, the N-acylethanolamine compound of meth FIG. 1 is a graph representing mean group ear Swelling (in ods and compositions of the present invention is palmitoy mm). Earthickness was measured prior to (baseline), and 3 15 lethanolamine (PEA) hours following, croton oil induced ear sensitization. The N-acylethanolamine compound can also be OEA, FIG. 2a is a graph representing left Earthickness (in mm) Me-PEA or PIA (Jonsson et al. Vandevoorde S. Lambert D 6 hours post sensitization. Thickness of sensitized ears was M, Tiger G, Fowler C.J. Br J Pharmacol. 2001; 133:1263 measured 24 hours prior to (baseline), and 6 hours follow 75): ing, croton oil induced ear sensitization FIG.2b is a graph representing mean left Earthickness (in mm) 6 hours post sensitization. Thickness of sensitized ears was measured 24 hours prior to (baseline), and 6 hours OH following, croton oil induced ear sensitization 1N1 FIG. 3 is a graph representing left ear thickness (in mm) 25 H 6 hours and 9 hours post sensitization. DESCRIPTION OF THE PREFERRED Palmitoylethanolamide (PEA) EMBODIMENTS O 30 The present invention is of compositions which can be 1N1OH used to treat inflammatory disorders. The principles and operation of the present invention may be better understood with reference to the drawings and Oleoylethanolamide (OEA) accompanying descriptions. 35 Before explaining at least one embodiment of the inven tion in detail, it is to be understood that the invention is not limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being 40 cr practiced or carried out in various ways. Also, it is to be understood that the phraseology and terminology employed Palmitoylisopropylamide (PIA) herein is for the purpose of description and should not be regarded as limiting. Inflammatory disorders are typically treated with corti 45 costeroids which can lead to serious side effects which mimic Cushing's disease, a malfunction of the adrenal cirr glands resulting in an overproduction of cortisol. Other R-palmitoyl-(2-methyl)ethanolamide potential side effects associated with use of corticosteroids (Me-PEA) include increased appetite and weight gain, deposits offat in 50 chest, face, upper back, and stomach and water and salt retention leading to Swelling and edema. Thus, there is a The N-acylethanolamine compound can have a side chain great need to decrease the use of corticosteroids in treatment length of 16 carbon units. Alternatively, the N-acyletha of inflammatory disorders. nolamine compound can be decanoylethanolamide (C10:0) The present inventors have devised compositions which 55 lauroylethanolamide (C12:0), or myristoylethanolamide include a corticosteroid and a second compound which (C14:0). synergistically complements the corticosteroid in treatment The N-acylethanolamine compound can be an N-15 acy of inflammatory disorders. As a result of this synergism, the lethanolamine derivative. The ethanolamide group of the present compositions can be used to treat inflammatory N-acylethanolamine compound can be replaced with a moi disorders with substantially lower concentrations of corti 60 ety Such as, butylamide, isopro-pylamide, cyclohexamide costeroids or with low potency corticosteroids, thereby and (2-methyl)ethanolamide. Each possibility represents a augmenting the therapeutic properties of steroids by increas separate embodiment of the present invention. ing its therapeutic activity and/or reducing steroid associated The N-acylethanolamine compound can be a palmitoyle adverse effects. As a result the present compositions provide thanolamine derivative. The ethanolamide group of palmi a Substantial corticosteroid sparing effect. 65 toylethanolamine can be replaced with a moiety such as Thus according to one aspect of the present invention butylamide, isopropylamide, cyclohexamide and (2-methyl) there is provided a composition which includes a corticos ethanolamide. The N-acylethanolamine compound can be, US 9,623,000 B2 7 8 for example, palmitoylcyclohexamide, palmitoylbutylam A non-limiting example of a very-high-potency topical ide, palmitoylisopropylamide, palmitoylethanolamide, or corticosteroid is clobetasol-17-propionate (available as a 5 Me-palmitoylethanolamide. Each possibility represents a 5 0.05% cream under the name DermovateR). The corticos separate embodiment of the present invention. teroid of the compositions of the present invention can The N-acylethanolamine compound of the composition of 5 exhibit glucocorticoid activity and/or mineralocorticoid the present invention can be, for example, oleoylethanol activity. The corticosteroid can be, for example, mometa amide (OEA) or a derivative thereof. The ethanolamide Sone and a salt thereof, hydrocortisone, hydrocortisone group of OEA can be replaced with a moiety Such as, acetate, hydrocortisone butyrate, hydrocortisone sodium butylamide, isopro-pylamide, cyclohexamide or (2-methyl) phosphate, hydrocortisone sodium Succinate, hydrocorti ethanolamide. 10 Sone cypionate, hydrocortisone probutate, and hydrocorti Sone Valerate. OEA, its derivatives, and methods for synthesizing same The corticosteroid can also be, for example, dexametha are well known in the art, and are described, inter alia, in Sone, betamethasone, triamcinolone, triamcinolone U.S. Pat. Nos. 6,656,972 and 7,348,338 and United States acetonide, triamcinolone diacetate, triamcinolone hexac patent application 2002/0173550, which are incorporated 15 etonide, beclomethasone, dipropionate, beclomethasone herein by reference. dipropionate monohydrate, flumethasone pivalate, diflora Methods for synthesizing N-acylethanolamine com Sone diacetate, fluocinolone acetonide, fluorometholone, pounds are well known in the art. As described in Lambert fluorometholone acetate, clobetasol propionate, desoxi et al. PEA was initially synthesized by refluxing etha methasone, fluoxymesterone, fluprednisolone, cortisone nolamine with palmitic acid (Roe et al., J Am Chem Soc acetate, paramethasone 25 acetate, methylprednisolone, 1952, 74:3442), yielding white crystals melting at 98-99 C. methylprednisolone acetate, methylprednisolone sodium Due to the simplicity of structure, various syntheses of PEA Succinate, prednisolone, prednisolone acetate, prednisolone have been described: the acyl chloride is the most common, Sodium phosphate, prednisolone tebutate, clocortolone piva but activating agents such as dicyclohexylcarbodiimide and late, flucinolone, dexamethasone 21-acetate, betamethasone carbonyldiimidazole allow the condensation between the 25 17-valerate, isoflupredone, 9-fluorocortisone, 6-hydroxy acid and the ethanolamine in very good yields (>80%). dexamethasone, dichlorisone, meclorisone, flupredidene, As is described in the Examples section which follows, doxibetasol, halopredone, halometasone, 30 clobetaSone, the present inventors have shown, for the first time, that diflucortolone, isoflupredone acetate, fluorohydroxyandros N-acylethanolamine compounds exhibit a steroid-sparing tenedione, beclomethasone, flumethasone, diflorasone, fluo effect. Such effects enable use of a low-potency steroid in 30 cinolone, clobetasol, cortisone, paramethasone, clocor instances wherein a mid-potency or high-potency steroid is tolone, prednisolone 21-hemisuccinate free acid, typically required or use of lower doses of a high potency prednisolone metasulphobenzoate, prednisolone terbutate, steroid, in addition to mitigating Steroid associated cutane and triamcinolone acetonide 21-palmitate. ous adverse effects. Each possibility represents a separate The corticosteroid can be any naturally occurring or embodiment of the present invention. Corticosteroid and 35 synthetic steroid hormone that can be chemically derived N-acylethanolamine compounds useful in the invention from cholesterol and is characterized by a hydrogenated include those described herein in any of their pharmaceuti cyclopentanoperhydrophenanthrene ring system. Naturally cally acceptable forms, including isomers such as diaste occurring corticosteriods are typically generally produced reomers and enantiomers, salts, Solvates, and polymorphs by the adrenal cortex. Synthetic corticosteriods may be thereof, as well as racemic mixtures of the compounds 40 halogenated. Functional groups required for activity include described herein. a double bond at a C3 ketone, and a C20 ketone. Corticosteroids that form a part of the present composi The compositions of the present invention can be formu tion include low to very high potency corticosteroids. lated into a pharmaceutical composition. Non-limiting examples of low-potency topical corticos As used herein, a “pharmaceutical composition” refers to teroids include hydrocortisone (available as 0.5% and 1% 45 a preparation of the active ingredients described herein with cream under the names Cortate(R), Unicort(R), and Cor other chemical components such as physiologically Suitable tisporinR). carriers and excipients. The purpose of a pharmaceutical Nonlimiting examples of mid-potency topical corticoster composition is to facilitate administration of a compound to oids include betamethasone Valerate, (available as 0.05% an organism. and 0.1% cream under the names Betnovate(R) and Celesto 50 Hereinafter, the phrases “physiologically acceptable car derm(R), clobetasone-17-butyrate (available as 0.05% cream rier' and “pharmaceutically acceptable carrier, which may under the name EumovateR), Flucinonide (available as be used interchangeably, refer to a carrier or a diluent that 0.05% and 0.01% cream under the names 15 Lidex(R) and does not cause significant irritation to an organism and does Lidemol(R), fluocinolone acetonide (available as 0.025% not abrogate the biological activity and properties of the and 0.01% cream under the names Synalar R, Synamol(R), 55 administered composition. An adjuvant is included under and Derma-Smooth R), and triamcinolone acetonide (avail these phrases. able as 0.025% and 0.1% cream under the names Aristocort Herein, the term "excipient” refers to an inert substance DR), Aristocort RR, Kenalog(R), and Vicoderm-KC(R). added to a pharmaceutical composition to further facilitate Non-limiting examples of high-25 potency topical corti administration of an active ingredient. Examples, without costeroids include betamethasone-17-benzoate (available as 60 limitation, of excipients include calcium carbonate, calcium a 0.025% cream under the name Beben(R), betamethasone phosphate, various Sugars and types of starch, cellulose dipropionate (available as a 0.025% cream under the name derivatives, gelatin, vegetable oils, and polyethylene gly Propaderm R) and as a 0.05% cream under the names cols. Diprosone(R) and Diprolene GlycolR), halcinonide (avail Techniques for formulation and administration of drugs able as a 0.1% cream under the name Halog(R), and triam 65 may be found in the latest edition of “Remington’s Phar cinolone acetonide (available as a 0.5% cream under the maceutical Sciences.” Mack Publishing Co., Easton, Pa., name 30 Aristocort C(R). which is herein fully incorporated by reference. US 9,623,000 B2 10 Suitable routes of administration may, for example, binders such as starches, lubricants such as talc or magne include topical, oral, rectal, transmucosal, especially trans sium Stearate, and, optionally, stabilizers. In soft capsules, nasal, intestinal, or parenteral delivery, including intramus the active ingredients may be dissolved or Suspended in cular, Subcutaneous, and intramedullary injections, as well Suitable liquids, such as fatty oils, liquid paraffin, or liquid as intrathecal, direct intraventricular, intravenous, intraperi polyethylene glycols. In addition, stabilizers may be added. toneal, intranasal, or intraocular injections. All formulations for oral administration should be in dos Alternately, one may administer the pharmaceutical com ages Suitable for the chosen route of administration. position in a local rather than systemic manner, for example, For buccal administration, the compositions may take the via injection of the pharmaceutical composition directly into form of tablets or lozenges formulated in conventional a tissue region of a patient. 10 manner or in adhesive carriers. Pharmaceutical compositions of the present invention For administration by inhalation, the active ingredients may be manufactured by processes well known in the art, for use according to the present invention are conveniently e.g., by means of conventional mixing, dissolving, granu delivered in the form of an aerosol spray presentation from lating, dragee-making, levigating, emulsifying, encapsulat a pressurized pack or a nebulizer with the use of a suitable ing, entrapping, or lyophilizing processes. 15 propellant, e.g., dichlorodifluoromethane, trichlorofluo Pharmaceutical compositions for use in accordance with romethane, dichloro-tetrafluoroethane, or carbon dioxide. In the present invention thus may be formulated in conven the case of a pressurized aerosol, the dosage may be deter tional manner using one or more physiologically acceptable mined by providing a valve to deliver a metered amount. carriers comprising excipients and auxiliaries, which facili Capsules and cartridges of, for example, gelatin for use in a tate processing of the active ingredients into preparations dispenser may be formulated containing a powder mix of the that can be used pharmaceutically. Proper formulation is compound and a suitable powder base. Such as lactose or dependent upon the route of administration chosen. starch. For topical, the active ingredients of the pharmaceutical The pharmaceutical composition described herein may be composition may be formulated in cremes, ointments, solu formulated for parenteral administration, e.g., by bolus tions, patches, sprays, lotions, liniments, varnishes, Solid 25 injection or continuous infusion. Formulations for injection preparations such as silicone sheets, and the like. may be presented in unit dosage form, e.g., in ampoules or For injection, the active ingredients of the pharmaceutical in multidose containers with, optionally, an added preserva composition may be formulated in aqueous solutions, pref tive. The compositions may be suspensions, solutions, or erably in physiologically compatible buffers such as Hank’s emulsions in oily or aqueous vehicles, and may contain Solution, Ringer's Solution, or physiological salt buffer. For 30 formulatory agents such as Suspending, stabilizing, and/or transmucosal administration, penetrants appropriate to the dispersing agents. barrier to be permeated are used in the formulation. Such Pharmaceutical compositions for parenteral administra penetrants are generally known in the art. tion include aqueous solutions of the active preparation in For oral administration, the pharmaceutical composition water-soluble form. Additionally, suspensions of the active can be formulated readily by combining the active com 35 ingredients may be prepared as appropriate oily or water pounds with pharmaceutically acceptable carriers well based injection Suspensions. Suitable lipophilic solvents or known in the art. Such carriers enable the pharmaceutical vehicles include fatty oils such as sesame oil, or synthetic composition to be formulated as tablets, pills, dragees, fatty acid esters such as ethyl oleate, triglycerides, or lipo capsules, liquids, gels, syrups, slurries, Suspensions, and the Somes. Aqueous injection Suspensions may contain Sub like, for oral ingestion by a patient. Pharmacological prepa 40 stances that increase the Viscosity of the Suspension, such as rations for oral use can be made using a solid excipient, sodium carboxymethyl cellulose, sorbitol, or dextran. optionally grinding the resulting mixture, and processing the Optionally, the Suspension may also contain Suitable stabi mixture of granules, after adding Suitable auxiliaries as lizers or agents that increase the solubility of the active desired, to obtain tablets or dragee cores. Suitable excipients ingredients, to allow for the preparation of highly concen are, in particular, fillers such as Sugars, including lactose, 45 trated Solutions. Sucrose, mannitol, or Sorbitol; cellulose preparations such Alternatively, the active ingredient may be in powder as, for example, maize starch, wheat starch, rice starch, form for constitution with a Suitable vehicle, e.g., a sterile, potato starch, gelatin, gum tragacanth, methyl cellulose, pyrogen-free, water-based solution, before use. hydroxypropylmethyl-cellulose, and Sodium carbomethyl The pharmaceutical composition of the present invention cellulose; and/or physiologically acceptable polymers such 50 may also be formulated in rectal compositions such as as polyvinylpyrrolidone (PVP). If desired, disintegrating Suppositories or retention enemas, using, for example, con agents, such as cross-linked polyvinyl pyrrolidone, agar, or ventional Suppository bases such as cocoa butter or other alginic acid or a salt thereof. Such as Sodium alginate, may glycerides. be added. The present compositions can also be delivered using an Dragee cores are provided with suitable coatings. For this 55 in-situ formed depot (ISFD). Examples of in-situ formed purpose, concentrated Sugar Solutions may be used which depots include semi-solid polymers which can be injected as may optionally contain gum arabic, talc, polyvinyl pyrroli a melt and form a depot upon cooling to body temperature. done, carbopol gel, polyethylene glycol, titanium dioxide, The requirements for such ISFD include low melting or lacquer Solutions, and Suitable organic solvents or solvent glass transition temperatures in the range of 25-658 C and an mixtures. Dyestuffs or pigments may be added to the tablets 60 intrinsic viscosity in the range of 0.05-0.8 dl/g 12-14. or dragee coatings for identification or to characterize dif Below the viscosity threshold of 0.05 dl/g no delayed ferent combinations of active compound doses. diffusion could be observed, whereas above 0.8 dl/g the Pharmaceutical compositions that can be used orally ISFD was no longer injectable using a needle. At injection include push-fit capsules made of gelatin, as well as Soft, temperatures above 378 C but below 658 C these polymers sealed capsules made of gelatin and a plasticizer, Such as 65 behave like viscous fluids which solidify to highly viscous glycerol or Sorbitol. The push-fit capsules may contain the depots. Drugs are incorporated into the molten polymer by active ingredients in admixture with filler such as lactose, mixing without the application of solvents. Thermoplastic US 9,623,000 B2 11 12 pastes (TP) can be used to generate a Subcutaneous drug Determination of a therapeutically effective amount is reservoir from which diffusion occurs into the systemic well within the capability of those skilled in the art, espe circulation. cially in light of the detailed disclosure provided herein. In situ cross-linked polymer systems utilize a cross-linked For any preparation used in the methods of the invention, polymer network to control the diffusion of macromolecules the dosage or the therapeutically effective amount can be over a prolonged period of time. Use of in situ cross-linking estimated initially from in vitro and cell culture assays. For implants necessitate protection of the bioactive agents dur example, a dose can be formulated in animal models to ing the cross-linking reaction. This could be achieved by achieve a desired concentration or titer. Such information encapsulation into fast degrading gelatin microparticles. can be used to more accurately determine useful doses in 10 humans. An ISFD can also be based on polymer precipitation. A The dosage of each compound of the claimed combina water-insoluble and biodegradable polymer is dissolved in a tions depends on several factors, including: the administra biocompatible organic solvent to which a drug is added tion method, the disease to be treated, the severity of the forming a solution or Suspension after mixing. When this disease, whether the disease is to be treated or prevented, formulation is injected into the body the water miscible 15 and the age, weight, and health of the person to be treated. organic solvent dissipates and water penetrates into the Additionally, pharmacogenomic (the effect of genotype on organic phase. This leads to phase separation and precipi the pharmacokinetic, pharmacodynamic or efficacy profile tation of the polymer forming a depot at the site of injection. of a therapeutic) information about a particular patient may One example of such a system is AtrigeleTM (ARTIX Labo affect dosage used. ratories). Continuous daily dosing may not be required; a therapeu Thermally induced gelling systems can also be used as tic regimen may require cycles, during which time a drug is ISFDs. Numerous polymers show abrupt changes in solu not administered, or therapy may be provided on an as bility as a function of environmental temperature. The needed basis during periods of acute inflammation. prototype of a thermosensitive polymer is poly(N-isopropyl Dosages for PEA and various steroids for topical appli acryl amide), poly-NIPAAM, which exhibits a rather sharp 25 cation are exemplified below. lower critical Solution temperature. Topical Steroid Group I Thermoplastic pastes Such as the new generation of PEA (0.3-5%)--Clobetasol diproprionate (0.01-0.05%) poly(ortho esters) developed by AP Pharma can also be used PEA (0.3-5%)--Betamethasone diproprionate 0.05-0.25% for depot drug delivery. Such pastes include polymers that PEA (0.3-5%)--Halbetasol proprionate 0.01-0.05% are semi-solid at room temperature, hence heating for drug 30 PEA (0.3-5%)--Diflorasone diacetate 0.01-0.05% incorporation and injection is no longer necessary. Injection Topical Steroid Group II is possible through needles no larger than 22 gauge. The PEA (0.3-5%)--Fluocinonide (0.01 0.05%) drug can be mixed into the systems in a dry and, therefore, PEA (0.3-5%)--Halcinonide (0.01-0.05%) stabilized state. Shrinkage or Swelling upon injection is PEA (0.3-5%)--Amcinonide (0.01 0.05%) thought to be marginal and, therefore, the initial drug burst 35 PEA (0.3-5%)--Desoximetasone (0.05-0.25%) is expected to be lower than in the other types of ISFD. An Topical Steroid Group III additional advantage is afforded by the self-catalyzed deg PEA (0.3-5%)--Triamcinalone acetonide (0.01-0.5%) radation by Surface erosion. PEA (0.3-5%)--Mometasone furoate (0.02-0.1%) The compositions of the present invention can also be PEA (0.3-5%)--Fluticasone proprionate (0.001-0.005%) delivered from medical devices, such as orthopedic 40 PEA (0.3-5%)--Betamethasone diproprionate (0.01-0.05%) implants, contact lenses, micro needle arrays, patches and Topical Steroid Group IV the like. PEA (0.3-5%)--Fluocinolone acetonide (0.05-0.2%) Sustained-release (SR), extended-release (ER, XR, or PEA (0.3-5%)--Hydrocortisone valerate (0.05-0.2%) XL), time-release or timed-release, controlled-release (CR), PEA (0.3-5%)--Hydrocortisone butyrate (0.02-0.1%) or continuous-release (CR or Contin) pills are tablets or 45 PEA (0.3-5%)--Flurandrenolide (0.01-0.05%) capsules formulated to dissolve slowly and release a drug PEA (0.3-5%)--Triamcinalone acetonide (0.02-0.1%) over time. Sustained-release tablets are formulated so that PEA (0.3-5%)--Mometasone furoate (0.02-0.1%) the active ingredient is embedded in a matrix of insoluble Topical Steroid Group V Substance (e.g. acrylics, polysaccharides etc) Such that the PEA (0.3-5%)--Triamcinalone acetonide (0.02-0.1%) dissolving drug diffuses out through the holes in the matrix. 50 PEA (0.3-5%)--Fluticasone propionate (0.01-0.05%) In some SR formulations the matrix physically swells up to PEA (0.3-5%)--Desonide (0.01-0.05%) form a gel, so that the drug has first to dissolve in matrix, PEA (0.3-5%)--Fluocinolone acetonide (0.005-0.025%) then exit through the outer surface. Difference between PEA (0.3-5%)--Hydrocortisone valerate (0.04-0.2%) controlled release and sustained release is that controlled Topical Steroid Group VI release is perfectly Zero order release that is, the drug 55 PEA (0.3-5%)--Prednicarbate (0.01-0.05%) releases with time irrespective of concentration. On the other PEA (0.3-5%)--Triamcinalone acetonide (0.005-0.025%) hand, Sustained release implies slow release of the drug over PEA (0.3-5%)--Fluocinolone acetonide (0.002-0.01%) a time period. It may or may not be controlled release. PEA (0.3-5%)--Desonide (0.01-0.05%) Pharmaceutical compositions suitable for use in the con Topical Steroid Group VII text of the present invention include compositions wherein 60 PEA (0.3-5%)--Hydrocortisone (0.5-2.5%) the active ingredients are contained in an amount effective to PEA (0.3-5%)--Hydrocortisone (0.2-1%) achieve the intended purpose. More specifically, a “thera An Inhalation formulation of a corticosteroid (ICS) and peutically effective amount’ means an amount of active PEA for treatment of for example, Asthma, COPD and other ingredients (e.g., a nucleic acid construct) effective to pre steroid responsive respiratory diseases and conditions can vent, alleviate, or ameliorate symptoms of a disorder (e.g., 65 include a dose of PEA of about 1 microgram to about 500 ischemia) or prolong the Survival of the Subject being milligrams per kilogram of the Subject, about 100 micro treated. grams to about 200 milligrams per kilogram of the Subject, US 9,623,000 B2 13 14 about 100 micrograms to about 5 milligrams per kilogram of individual characteristics and route of administration. Detec the Subject, or about 200 micrograms to about 2 milligrams tion assays can be used to determine plasma concentrations. per kilogram of the Subject. Depending on the severity and responsiveness of the The composition can be delivered by dispenser, wherein condition to be treated, dosing can be of a single or a the dispenser delivers a dose for inhalation of PEA of 1 5 plurality of administrations, with course of treatment lasting microgram to about 500 milligrams per kilogram of a from several days to several weeks, or until cure is effected Subject in need of asthma treatment. or diminution of the disease state is achieved. As an inhalant, PEA can be delivered in combination with The amount of a composition to be administered will, of any of the following steroids: course, be dependent on the Subject being treated, the 10 severity of the affliction, the manner of administration, the Beclometasone dipropionate (BDP) 40 to 320 mcg twice judgment of the prescribing physician, etc. daily Compositions of the present invention may, if desired, be Budesonide (BUD) Budesonide 180 to 720 mcg twice presented in a pack or dispenser device, such as an FDA daily approved kit, which may contain one or more unit dosage Ciclesonide (CIC): 80-320 mcg twice daily. 15 forms containing the active ingredient. The pack may, for Fluticasone propionate (FP) 100-500 mcg twice daily. example, comprise metal or plastic foil. Such as a blister Mometasone furoate (MF) 220-440 mcg once daily. pack. The pack or dispenser device may be accompanied by A nasal spray formulation of the present composition can instructions for administration. The pack or dispenser device include, for example, a dose of PEA of about 1 microgram may also be accompanied by a notice in a form prescribed to about 500 milligrams per kilogram of the subject, about by a governmental agency regulating the manufacture, use, 100 micrograms to about 200 milligrams per kilogram of the or sale of pharmaceuticals, which notice is reflective of Subject, about 100 micrograms to about 5 milligrams per approval by the agency of the form of the compositions for kilogram of the Subject, or about 200 micrograms to about human or veterinary administration. Such notice, for 2 milligrams per kilogram of the Subject. example, may include labeling approved by the U.S. Food As a nasal formulation, PEA can be delivered in combi 25 and Drug Administration for prescription drugs or of an nation with the following steroids: approved product insert. Compositions comprising a prepa Beclomethasone dipropionate, monohydrate, Nasal spay, ration of the invention formulated in a pharmaceutically (84-336 mcg/day). acceptable carrier may also be prepared, placed in an appro fluticasone propionate Nasal Spray, (50-200 mcg/day). priate container, and labeled for treatment of an indicated Triamcinolone Acetonide (110-220 mcg per day 30 inflammatory disorder, as further detailed above. The present composition can also be formulated for According to another aspect of the present invention there ophthalmic delivery (e.g. as drops) for treatment of ophthal is provided a method of treating an inflammatory disorder. mic diseases. The method is effected by administering to a subject in need An ophthalmic formulation of the present composition a therapeutically effective amount of the present composi can include: 35 tion. As used herein, the phrase “subject in need refers to PEA (0.3-5%)--loteprednoletabonate ophthalmic suspen a mammal, preferably a human that is Suffering from or is sion, 0.2%); predisposed to an inflammatory disorder. PEA (0.3-5%)--Betamethasone sodium phosphate (0.05 Local and systemic inflammatory disorders can be treated 0.1%); or with the pharmaceutical composition of the present inven PEA (0.3-5%)--Dexamethasone sodium phosphate 0.05 40 tion. Examples of local inflammatory disorders include, but O.1% are not limited to, asthma, gastrointestinal inflammation, Aural preparations of the present invention can include: ocular inflammation and cutaneous inflammatory disorder. PEA (0.3-5%)--Betamethasone sodium phosphate (0.05 Gastrointestinal inflammation can be caused by gastritis, 0.1%); or enteritis, proctitis, inflammatory bowel disease, Crohn dis PEA (0.3-5%)--Dexamethasone sodium phosphate 0.05 45 ease (CD) or ulcerative colitis (UC). Ocular inflammation O.1% can be caused by uvetis or iritis. Cutaneous inflammatory One skilled in the art will be able to ascertain suitable disorder can be caused by psoriasis, atopic dermatitis, and dosages for other steroid compounds for any of the above Scleroderma. described formulations. The present invention also envisages use of a corticos Toxicity and therapeutic efficacy of the active ingredients 50 teroid along with a second active agent Such as a tricyclic described herein can be determined by standard pharmaceu antidepressant, a GABA agonist, a monoamine oxidase tical procedures in vitro, in cell cultures or experimental inhibitor, a cannabinoid receptor agonist, and an immuno animals. The data obtained from these in vitro and cell philin inhibitor. The beneficial effects of such second active culture assays and animal studies can be used in formulating agent enable use of a low-potency Steroid in instances a range of dosage for use in human. The dosage may vary 55 wherein a mid-potency or high-potency Steroid is typically depending upon the dosage form employed and the route of required, use of a mid-potency steroid in instances wherein administration utilized. The exact formulation, route of a high-potency Steroid is typically required or use of lower administration, and dosage can be chosen by the individual doses of a high-potency steroid. physician in view of the patients condition. (See, e.g., Fingl, Thus, according to another aspect of the present invention E. et al. (1975), “The Pharmacological Basis of Therapeu 60 there is provided a composition-of-matter which includes a tics.' Ch. 1, p. 1.) corticosteroid and at least one additional active ingredient Dosage amount and administration intervals may be selected form the group consisting of a tricyclic antidepres adjusted individually to provide sufficient plasma or brain sant, a GABA agonist, a monoamine oxidase inhibitor, a levels of the active ingredient to suppress inflammation (i.e., cannabinoid receptor agonist, and an immunophilin inhibi minimally effective concentration, MEC). The MEC will 65 tOr. vary for each preparation, but can be estimated from in vitro The tricyclic antidepressant, GABA agonist, monoamine data. Dosages necessary to achieve the MEC will depend on oxidase inhibitor, and cannabinoid receptor agonist com US 9,623,000 B2 15 16 pounds useful in the invention include those described which are incorporated herein by reference in their entirety. hereinunder in any of their pharmaceutically acceptable Each possibility represents a separate embodiment of the forms, including isomers such as diastereomers and present invention. enantiomers, salts, Solvates, and polymorphs thereof, as well In another embodiment, “tricyclic antidepressant refers as racemic mixtures of the compounds described herein. A to a compound having one of the formulas: having one the corticosteroid suitable for use with this aspect of the present formulas (I), (II), or (III): invention is described hereinabove. Compositions including any of the combinations described above can be formulated as for topical adminis (I) tration for treatment of a cutaneous inflammatory disorder 10 X X that otherwise (in the absence of the tricyclic antidepressant) X Y. X requires a mid-potency steroid or high-potency steroid). In another embodiment, the tricyclic antidepressant and corti costeroid are the sole active ingredients in the composition. 15 X Z X In another embodiment, one or more additional active ingre X A X dients are present. In another embodiment, the tricyclic a N(B) antidepressant and corticosteroid are present in amounts (II) that, when administered together to a patient having cuta X X neous inflammatory disease or disorder, inhibit or reduce inflammation caused therefrom. In another embodiment, the X Y X active ingredients are present in amounts that treat the disorder. Each possibility represents a separate embodiment of the present invention. X X In another embodiment, the present invention provides a 25 X A X method for treating a cutaneous inflammatory disease or SN(B), disorder in a Subject in need thereof, the method comprising (III) the step of topically administering to the Subject a compo X X sition comprising a tricyclic antidepressant and a low potency corticosteroid, thereby treating a cutaneous inflam 30 matory disease or disorder. In another embodiment, the cutaneous inflammatory disease or disorder is a disease or disorder that otherwise (in the absence of the tricyclic antidepressant) requires a mid-potency Steroid or high 35 potency steroid). In another embodiment, a method of the present invention enables use of a mid-potency steroid in situations wherein a high-potency steroid would otherwise be required. In another embodiment, the method is used to alleviate a cutaneous inflammatory disease or disorder in a 40 subject in need thereof. In another embodiment, the topical administration is performed at a site afflicted by the inflam wherein each X is, independently, H, Cl, F, Br, I, CH, matory disorder. In another embodiment, the tricyclic anti CF, OH, OCH, CHCH, or OCHCH;Y is CH, O, NH, depressant and corticosteroid are the sole active ingredients S(O) (CH), (CH), CHO, CH-NH, CHN, or in the composition. In another embodiment, one or more 45 CHS: Z is C or S.; A is a branched or unbranched, additional active ingredients are present. In another embodi saturated or monounsaturated hydrocarbon chain having ment, the tricyclic antidepressant and corticosteroid are between 3 and 6 carbons, inclusive; each B is, indepen present in the composition in amounts that, when adminis dently, H, Cl, F, Br, I, CX CHCH, OCX, or OCX2CX: tered together to a patient having cutaneous inflammatory and D disease or disorder, inhibit or reduce inflammation caused 50 is CH, O, NH, S(O). therefrom. In another embodiment, the active ingredients are In another embodiment, each X is, independently, H., Cl, present in amounts that treat the disorder. Each possibility or F: Y is (CH), Z is C; A is (CH); and each B is, represents a separate embodiment of the present invention. independently, H., Cl, or F. The tricyclic antidepressant of methods and compositions In another embodiment of methods and compositions of of the present invention is, in Some preferred embodiments, 55 the present invention, a topical composition comprises an amitriptyline (available as (Elavil(R). Non-limiting examples additional active ingredient besides the tricyclic antidepres of tricyclic antidepressants useful in methods and composi sant and corticosteroid. In another embodiment, more than tions of the present invention are amoxapine, 8-hy one additional active ingredients are present. In another droxyamoxapine, 7-hydroxyamoxapine, loxapine, loxapine embodiment, two additional active ingredients are present. Succinate, loxapine hydrochloride, 8-hydroxyloxapine, dox 60 In another embodiment, more than two additional active epin, maprotiline, clomipramine, desipramine, imipramine, ingredients are present. Each possibility represents a sepa trimipramine, nortriptyline, and protriptyline. In another rate embodiment of the present invention. embodiment, the tricyclic antidepressant is any other tricy In another embodiment, the present invention provides a clic antidepressant known in the art. Tricyclic antidepres composition formulated for topical administration, the com sants are well known in the art, and are described in, for 65 position comprising a GABA agonist and a corticosteroid. In example, U.S. Pat. Nos. 2,554,736, 3,438,981, 3,420,851, another embodiment, the topical administration is in situ 3,534,041, 3,505,321, 4,013,639, 4,105,785, 6,368,814, topical administration. In another embodiment, the topical US 9,623,000 B2 17 18 administration is in situ topical administration. In another which are incorporated herein by reference in their entirety. embodiment, the corticosteroid is a low-potency corticos Each possibility represents a separate embodiment of the teroid. In another embodiment, the composition is indicated present invention. for a cutaneous inflammatory disease or disorder that oth In another embodiment, the present invention provides a erwise (in the absence of the GABA agonist) requires a composition formulated for topical administration, the com mid-potency steroid or high-potency steroid). In another position comprising a carbonic anhydrase inhibitor and a embodiment, the GABA agonist and corticosteroid are the corticosteroid. In another embodiment, the corticosteroid is sole active ingredients in the composition. In another a low-potency corticosteroid. In another embodiment, the embodiment, one or more additional active ingredients are cutaneous inflammatory disease or disorder is a disease or 10 disorder that otherwise (in the absence of the carbonic present. In another embodiment, the GABA agonist and anhydrase inhibitor) requires a mid-potency steroid or high corticosteroid are present in amounts that, when adminis potency steroid). In another embodiment, a method of the tered together to a patient having cutaneous inflammatory present invention enables use of a mid-potency steroid in disease or disorder, inhibit or reduce inflammation caused situations wherein a high-potency steroid would otherwise therefrom. In another embodiment, the active ingredients are 15 be required. In another embodiment, the topical administra present in amounts that treat the disorder. Each possibility tion is in situ topical administration. In another embodiment, represents a separate embodiment of the present invention. the carbonic anhydrase inhibitor and corticosteroid are the In another embodiment, the present invention provides a sole active ingredients in the composition. In another method for treating a cutaneous inflammatory disease or embodiment, one or more additional active ingredients are disorder in a Subject in need thereof, the method comprising present. In another embodiment, the carbonic anhydrase the step of topically administering to the Subject a compo inhibitor and corticosteroid are present in amounts that, sition comprising a GABA agonist and a corticosteroid, when administered together to a patient having cutaneous thereby treating a cutaneous inflammatory disease or disor inflammatory disease or disorder, inhibit or reduce inflam der. In another embodiment, the method is used to alleviate mation caused therefrom. In another embodiment, the active a cutaneous inflammatory disease or disorder in a Subject in 25 ingredients are present in amounts that treat the disorder. need thereof. In another embodiment, the topical adminis Each possibility represents a separate embodiment of the tration is in situ topical administration. In another embodi present invention. ment, the corticosteroid is a low-potency corticosteroid. In In another embodiment, the present invention provides a another embodiment, the cutaneous inflammatory disease or method for treating a cutaneous inflammatory disease or 30 disorder in a Subject in need thereof, the method comprising disorder is a disease or disorder that otherwise (in the the step of topically administering to the Subject a compo absence of the GABA agonist) requires a mid-potency sition comprising a carbonic anhydrase inhibitor and a steroid or high-potency steroid). In another embodiment, the corticosteroid, thereby treating a cutaneous inflammatory topical administration is performed at a site afflicted by the disease or disorder. In another embodiment, the method is inflammatory disorder. In another embodiment, the GABA 35 used to alleviate a cutaneous inflammatory disease or dis agonist and corticosteroid are the sole active ingredients in order in a subject in need thereof. In another embodiment, the composition. In another embodiment, one or more the topical administration is in situ topical administration. In additional active ingredients are present. In another embodi another embodiment, the corticosteroid is a low-potency ment, the GABA agonist and corticosteroid are present in the corticosteroid. In another embodiment, the cutaneous composition in amounts that, when administered together to 40 inflammatory disease or disorder is a disease or disorder that a patient having cutaneous inflammatory disease or disorder, otherwise (in the absence of the carbonic anhydrase inhibi inhibit or reduce inflammation caused therefrom. In another tor) requires a mid-potency steroid or high-potency steroid). embodiment, the active ingredients are present in amounts In another embodiment, the topical administration is per that treat the disorder. Each possibility represents a separate formed at a site afflicted by the inflammatory disorder. In embodiment of the present invention. 45 another embodiment, the carbonic anhydrase inhibitor and In another embodiment of methods and compositions of corticosteroid are the sole active ingredients in the compo the present invention, a topical composition comprises an sition. In another embodiment, one or more additional active additional active ingredient besides the GABA agonist and ingredients are present. In another embodiment, the carbonic corticosteroid. In another embodiment, more than one addi anhydrase inhibitor and corticosteroid are present in the tional active ingredients are present. In another embodiment, 50 composition in amounts that, when administered together to two additional active ingredients are present. In another a patient having cutaneous inflammatory disease or disorder, embodiment, more than two additional active ingredients are inhibit or reduce inflammation caused therefrom. In another present. Each possibility represents a separate embodiment embodiment, the active ingredients are present in amounts of the present invention. that treat the disorder. Each possibility represents a separate “GABA agonist” refers, in another embodiment, to an 55 embodiment of the present invention. activator of a GABA-gated chloride channel. In another The carbonic anhydrase inhibitor of methods and com embodiment, the GABA agonist of methods and composi positions of the present invention is, in some preferred tions of the present invention is, in some preferred embodi embodiments, topiramate. In another embodiment, the car ments, topiramate. Non-limiting examples of GABA ago bonic anhydrase inhibitor is a Sulfonamide-class carbonic nists useful in methods and compositions of the present 60 anhydrase inhibitor. Non-limiting examples of carbonic invention are topiramate, muscimol, progabide, riluzole, anhydrase inhibitors useful in methods and compositions of baclofen, gabapentin, vigabatrin, valproic acid, tiagabine, the present invention are topiramate and those compounds lamotrigine, pregabalin, phenyloin, and carbamazepine. In described in U.S. Pat. Nos. 4,929,637, 5,276,025, 5,464,831, another embodiment, the GABA agonist is any other GABA 5,473,067, 5,646,142, 7,030,250, 7,109,353, which are agonist known in the art. GABA agonists are well known in 65 incorporated herein by reference in their entirety. In another the art, and are described, for example, in U.S. Pat. Nos. embodiment, the carbonic anhydrase inhibitor is any other 5,006,560, 5,248,678, 6,077,839, 6,720,348, and 6,821,985, carbonic anhydrase inhibitor known in the art. Carbonic US 9,623,000 B2 19 20 anhydrase inhibitors are well known in the art, and are In another embodiment of methods and compositions of described, for example, in the above patents. Each possibil the present invention, a topical composition comprises an ity represents a separate embodiment of the present inven additional active ingredient besides the sulfamate-substi tion. tuted monosaccharide and corticosteroid. In another In another embodiment of methods and compositions of 5 embodiment, more than one additional active ingredients are the present invention, a topical composition comprises an present. In another embodiment, two additional active ingre additional active ingredient besides the carbonic anhydrase dients are present. In another embodiment, more than two inhibitor and corticosteroid. In another embodiment, more additional active ingredients are present. Each possibility than one additional active ingredients are present. In another represents a separate embodiment of the present invention. 10 In another embodiment, the present invention provides a embodiment, two additional active ingredients are present. composition formulated for topical administration, the com In another embodiment, more than two additional active position comprising a monoamine oxidase inhibitor ingredients are present. Each possibility represents a sepa (MAOI), and a corticosteroid. In another embodiment, the rate embodiment of the present invention. topical administration is in situ topical administration. In In another embodiment, the present invention provides a 15 another embodiment, the topical administration is in situ composition formulated for topical administration, the com topical administration. In another embodiment, the corticos position comprising a sulfamate-substituted monosaccha teroid is a low-potency corticosteroid. In another embodi ride and a corticosteroid. In another embodiment, the topical ment, the composition is indicated for a cutaneous inflam administration is in situ topical administration. In another matory disease or disorder that otherwise (in the absence of embodiment, the corticosteroid is a low-potency corticos the monoamine oxidase inhibitor) requires a mid-potency teroid. In another embodiment, the composition is indicated steroid or high-potency steroid). In another embodiment, the for a cutaneous inflammatory disease or disorder that oth monoamine oxidase inhibitor and corticosteroid are the sole erwise (in the absence of the sulfamate-substituted mono active ingredients in the composition. In another embodi saccharide) requires a mid-potency steroid or high-potency ment, one or more additional active ingredients are present. steroid). In another embodiment, a method of the present 25 In another embodiment, the monoamine oxidase inhibitor invention enables use of a mid-potency Steroid in situations and corticosteroid are present in amounts that, when admin wherein a high-potency steroid would otherwise be required. istered together to a patient having cutaneous inflammatory In another embodiment, the Sulfamate-substituted monosac disease or disorder, inhibit or reduce inflammation caused charide and corticosteroid are the sole active ingredients in therefrom. In another embodiment, the active ingredients are the composition. In another embodiment, one or more 30 present in amounts that treat the disorder. Each possibility additional active ingredients are present. In another embodi represents a separate embodiment of the present invention. ment, the sulfamate-substituted monosaccharide and corti In another embodiment, the present invention provides a costeroid are present in amounts that, when administered method for treating a cutaneous inflammatory disease or together to a patient having cutaneous inflammatory disease disorder in a Subject in need thereof, the method comprising or disorder, inhibit or reduce inflammation caused there 35 the step of topically administering to the Subject a compo from. In another embodiment, the active ingredients are sition comprising a monoamine oxidase inhibitor and a present in amounts that treat the disorder. Each possibility corticosteroid, thereby treating a cutaneous inflammatory represents a separate embodiment of the present invention. disease or disorder. In another embodiment, the method is In another embodiment, the present invention provides a used to alleviate a cutaneous inflammatory disease or dis method for treating a cutaneous inflammatory disease or 40 order in a subject in need thereof. In another embodiment, disorder in a Subject in need thereof, the method comprising the topical administration is in situ topical administration. In the step of topically administering to the Subject a compo another embodiment, the corticosteroid is a low-potency sition comprising a sulfamate-substituted monosaccharide corticosteroid. In another embodiment, the cutaneous and a corticosteroid, thereby treating a cutaneous inflam inflammatory disease or disorder is a disease or disorder that matory disease or disorder. In another embodiment, the 45 otherwise (in the absence of the monoamine oxidase inhibi method is used to alleviate a cutaneous inflammatory disease tor) requires a mid-potency steroid or high-potency steroid). or disorder in a subject in need thereof. In another embodi In another embodiment, the topical administration is per ment, the topical administration is in situ topical adminis formed at a site afflicted by the inflammatory disorder. In tration. In another embodiment, the corticosteroid is a low another embodiment, the monoamine oxidase inhibitor and potency corticosteroid. In another embodiment, the 50 corticosteroid are the sole active ingredients in the compo cutaneous inflammatory disease or disorder is a disease or sition. In another embodiment, one or more additional active disorder that otherwise (in the absence of the sulfamate ingredients are present. In another embodiment, the mono Substituted monosaccharide) requires a mid-potency steroid amine oxidase inhibitor and corticosteroid are present in the or high-potency steroid). In another embodiment, the topical composition in amounts that, when administered together to administration is performed at a site afflicted by the inflam 55 a patient having cutaneous inflammatory disease or disorder, matory disorder. In another embodiment, the Sulfamate inhibit or reduce inflammation caused therefrom. In another Substituted monosaccharide and corticosteroid are the sole embodiment, the active ingredients are present in amounts active ingredients in the composition. In another embodi that treat the disorder. Each possibility represents a separate ment, one or more additional active ingredients are present. embodiment of the present invention. In another embodiment, the Sulfamate-substituted monosac 60 The monoamine oxidase inhibitor of methods and com charide and corticosteroid are present in the composition in positions of the present invention is, in some preferred amounts that, when administered together to a patient hav embodiments, moclobemide. In another embodiment, the ing cutaneous inflammatory disease or disorder, inhibit or monoamine oxidase inhibitor is a monoamine oxidase A reduce inflammation caused therefrom. In another embodi inhibitor. Non-limiting examples of monoamine oxidase ment, the active ingredients are present in amounts that treat 65 inhibitors useful in methods and compositions of the present the disorder. Each possibility represents a separate embodi invention are clorgiline, iproclozide, iproniazid, isocarbox ment of the present invention. azid (Marplan), minaprine, nialamide, pargyline, phenelzine US 9,623,000 B2 21 22 (Nardil), rasagiline, selegiline (Eldepryl), toloxatone, tranyl another embodiment, the cutaneous inflammatory disease or cypromine (Parnate), furazolidone (Furoxone), and procar disorder is a disease or disorder that otherwise (in the bazine (Matulane). In another embodiment, the monoamine absence of the RIMA compound) requires a mid-potency oxidase inhibitor is any other monoamine oxidase inhibitor steroid or high-potency steroid). In another embodiment, the that increases endogenous concentrations of epinephrine topical administration is performed at a site afflicted by the (adrenaline). In another embodiment, the monoamine oxi inflammatory disorder. In another embodiment, the RIMA dase inhibitor is any other monoamine oxidase inhibitor that compound and corticosteroid are the sole active ingredients increases endogenous concentrations of norepinephrine (no in the composition. In another embodiment, one or more radrenaline). In another embodiment, the monoamine oxi additional active ingredients are present. In another embodi dase inhibitor is any other monoamine oxidase inhibitor that 10 ment, the RIMA compound and corticosteroid are present in increases endogenous concentrations of dopamine. In the composition in amounts that, when administered another embodiment, the monoamine oxidase inhibitor is together to a patient having cutaneous inflammatory disease any other monoamine oxidase inhibitor that increases or disorder, inhibit or reduce inflammation caused there endogenous concentrations of serotonin. In another embodi from. In another embodiment, the active ingredients are ment, the monoamine oxidase inhibitor is any other mono 15 present in amounts that treat the disorder. Each possibility amine oxidase inhibitor known in the art to exhibit mono represents a separate embodiment of the present invention. amine oxidase A inhibitory activity. Monoamine oxidase The RIMA compound of methods and compositions of the inhibitors are well known in the art, and are described in, for present invention is, in another embodiment, selected from example, U.S. Pat. Nos. 5,508,311, 5,792,799, 6,472,423, the group consisting of befloxatone, brofaromine, and and 6,569.470, which are incorporated herein by reference in cimoxatone. In another embodiment, the RIMA compound their entirety. Each possibility represents a separate embodi is a beta-carboline RIMA compound. In another embodi ment of the present invention. Each possibility represents a ment, the RIMA compound is harmaline). In another separate embodiment of the present invention. embodiment, the RIMA compound is any other RIMA In another embodiment of methods and compositions of compound known in the art to exhibit monoamine oxidase A the present invention, a topical composition comprises an 25 inhibitory activity. Each possibility represents a separate additional active ingredient besides the monoamine oxidase embodiment of the present invention. inhibitor and corticosteroid. In another embodiment, more In another embodiment of methods and compositions of than one additional active ingredients are present. In another the present invention, a topical composition comprises an embodiment, two additional active ingredients are present. additional active ingredient besides the RIMA compound In another embodiment, more than two additional active 30 and corticosteroid. In another embodiment, more than one ingredients are present. Each possibility represents a sepa additional active ingredients are present. In another embodi rate embodiment of the present invention. ment, two additional active ingredients are present. In In another embodiment, the present invention provides a another embodiment, more than two additional active ingre composition formulated for topical administration, the com dients are present. Each possibility represents a separate position comprising a reversible inhibitor of monoamine 35 embodiment of the present invention. oxidase A (RIMA compound) and a corticosteroid. In In another embodiment, the present invention provides a another embodiment, the topical administration is in situ composition formulated for topical administration, the com topical administration. In another embodiment, the topical position comprising a cannabinoid receptor agonist and a administration is in situ topical administration. In another corticosteroid. In another embodiment, the topical adminis embodiment, the corticosteroid is a low-potency corticos 40 tration is in situ topical administration. In another embodi teroid. In another embodiment, the composition is indicated ment, the topical administration is in situ topical adminis for a cutaneous inflammatory disease or disorder that oth tration. In another embodiment, the topical administration is erwise (in the absence of the RIMA compound) requires a in situ topical administration. In another embodiment, the mid-potency steroid or high-potency steroid). In another corticosteroid is a low-potency corticosteroid. In another embodiment, the monoamine oxidase inhibitor and corticos 45 embodiment, the composition is indicated for a cutaneous teroid are the sole active ingredients in the composition. In inflammatory disease or disorder that otherwise (in the another embodiment, one or more additional active ingre absence of the cannabinoid receptor agonist) requires a dients are present. In another embodiment, the monoamine mid-potency steroid or high-potency steroid). In another oxidase inhibitor and corticosteroid are present in amounts embodiment, the cannabinoid receptor agonist and corticos that, when administered together to a patient having cuta 50 teroid are the sole active ingredients in the composition. In neous inflammatory disease or disorder, inhibit or reduce another embodiment, one or more additional active ingre inflammation caused therefrom. In another embodiment, the dients are present. In another embodiment, the cannabinoid active ingredients are present in amounts that treat the receptor agonist and corticosteroid are present in amounts disorder. Each possibility represents a separate embodiment that, when administered together to a patient having cuta of the present invention. 55 neous inflammatory disease or disorder, inhibit or reduce In another embodiment, the present invention provides a inflammation caused therefrom. In another embodiment, the method for treating a cutaneous inflammatory disease or active ingredients are present in amounts that treat the disorder in a Subject in need thereof, the method comprising disorder. Each possibility represents a separate embodiment the step of topically administering to the Subject a compo of the present invention. sition comprising a reversible inhibitor of monoamine oxi 60 In another embodiment, the present invention provides a dase A (RIMA compound) and a corticosteroid, thereby method for treating a cutaneous inflammatory disease or treating a cutaneous inflammatory disease or disorder. In disorder in a Subject in need thereof, the method comprising another embodiment, the method is used to alleviate a the step of topically administering to the Subject a compo cutaneous inflammatory disease or disorder in a Subject in sition comprising a cannabinoid receptor agonist and a need thereof. In another embodiment, the topical adminis 65 corticosteroid, thereby treating a cutaneous inflammatory tration is in situ topical administration. In another embodi disease or disorder. In another embodiment, the method is ment, the corticosteroid is a low-potency corticosteroid. In used to alleviate a cutaneous inflammatory disease or dis US 9,623,000 B2 23 24 order in a subject in need thereof. In another embodiment, CBD) receptor. In another embodiment, the cannabinoid the topical administration is in situ topical administration. In receptor agonist targets an anandamide receptor. Each pos another embodiment, the corticosteroid is a low-potency sibility represents a separate embodiment of the present corticosteroid. In another embodiment, the cutaneous invention. inflammatory disease or disorder is a disease or disorder that In another embodiment of methods and compositions of otherwise (in the absence of the cannabinoid receptor ago the present invention, a topical composition comprises an nist) requires a mid-potency steroid or high-potency Ste additional active ingredient besides the cannabinoid receptor agonist and corticosteroid. In another embodiment, more roid). In another embodiment, the topical administration is than one additional active ingredients are present. In another performed at a site afflicted by the inflammatory disorder. In embodiment, two additional active ingredients are present. another embodiment, the cannabinoid receptor agonist and 10 In another embodiment, more than two additional active corticosteroid are the sole active ingredients in the compo ingredients are present. Each possibility represents a sepa sition. In another embodiment, one or more additional active rate embodiment of the present invention. ingredients are present. In another embodiment, the can In another embodiment, the present invention provides a nabinoid receptor agonist and corticosteroid are present in composition formulated for topical administration, the com the composition in amounts that, when administered 15 position comprising an immunophilin inhibitor and a corti together to a patient having cutaneous inflammatory disease costeroid. In another embodiment, the topical administration or disorder, inhibit or reduce inflammation caused there is in situ topical administration. In another embodiment, the from. In another embodiment, the active ingredients are topical administration is in situ topical administration. In present in amounts that treat the disorder. Each possibility another embodiment, the topical administration is in situ represents a separate embodiment of the present invention. topical administration. In another embodiment, the corticos The cannabinoid receptor agonist of methods and com teroid is a low-potency corticosteroid. In another embodi positions of the present invention is, in Some preferred ment, the composition is indicated for a cutaneous inflam embodiments, cannabidiol (CBD). In another preferred matory disease or disorder that otherwise (in the absence of embodiment, the cannabinoid receptor agonist is an endo the immunophilin inhibitor) requires a mid-potency steroid cannaboid compound. Non-limiting examples of cannabi 25 or high-potency steroid). In another embodiment, the immu noid receptor agonists useful in methods and compositions nophilin inhibitor and corticosteroid are the sole active ingredients in the composition. In another embodiment, one of the present invention are cannabidivarol (CBDV), can or more additional active ingredients are present. In another nabinol (CBN), cannabigerol (CBG), cannabivarol (CBV), embodiment, the immunophilin inhibitor and corticosteroid cannabicyclol (CBL), tetrahydrocannabinol (THC), tetrahy are present in amounts that, when administered together to drocannabinol-C4. (THC-C4), tetrahydrocannabivarin (also 30 a patient having cutaneous inflammatory disease or disorder, known as tetrahydrocannabivarol, THCV, and THV), inhibit or reduce inflammation caused therefrom. In another 11-Hydroxy-A-tetrahydrocannabinol, (11-OH-THC), embodiment, the active ingredients are present in amounts 11-nor-9-Carboxy-A9-tetrahydrocannabinol (11-COOH that treat the disorder. Each possibility represents a separate THC, THC-COOH, THC-11-oic acid), arachidonoyl etha embodiment of the present invention. nolamide (Anandamide, AEA), 2-arachidonoylglycerol 35 In another embodiment, the present invention provides a (2-AG), 2-Arachidonyl glyceryl ether (noladin ether), virod method for treating a cutaneous inflammatory disease or hamine, N-arachidonoyl-dopamine (NADA), oleamide, disorder in a Subject in need thereof, the method comprising A-41988, ajulemic acid, AM-087, AM-411, AM-855, the step of topically administering to the Subject a compo AM-905, AM-906, AM-919, AM-938, AM-4030, AMG-1, sition comprising an immunophilin inhibitor and a corticos AMG-3, AMG-36, AMG-41, dexanabinol (HU-211), 40 teroid, thereby treating a cutaneous inflammatory disease or DMHP dronabinol, HU-210, JWH-051, JWH-133, JWH disorder. In another embodiment, the method is used to 139, L-759,633, L-759,656, levonantradol, nabilone, Nabi alleviate a cutaneous inflammatory disease or disorder in a tan, O-806, O-823, O-1057, O-1125, O-1238, O-2545, subject in need thereof. In another embodiment, the topical O-2694, parahexyl, THC-O-acetate, THC-O-phosphate. CP administration is in situ topical administration. In another 47,497, CP 55.244, CP 55,940, HU-308, 2-Isopropyl-5- 45 embodiment, the corticosteroid is a low-potency corticos methyl-1-(2,6-dihydroxy-4-nonylphenyl)cyclohex-1-ene, teroid. In another embodiment, the cutaneous inflammatory AM-630, AM-1241, JWH-015, JWH-018, JWH-073, JWH disease or disorder is a disease or disorder that otherwise (in 081, JWH-200, L-768,242, pravadoline, WIN 55, 212-2, the absence of the immunophilin inhibitor) requires a mid JWH-030, JWH-147, JWH-307, AM-883, Arachidonyl-2'- potency steroid or high-potency steroid). In another embodi chloroethylamide, arachidonylcyclopropylamide, meth 50 ment, the topical administration is performed at a site anandamide, O-585, O-689, O-1812, O-1860, O-1861, BAY afflicted by the inflammatory disorder. In another embodi 38-7271, BAY 59-3074, JWH-171, and O-2220. Cannabi ment, the immunophilin inhibitor and corticosteroid are the noid receptor agonists are well known in the art, and are sole active ingredients in the composition. In another described in, for example, U.S. Pat. Nos. 5,747,524, 5,596, embodiment, one or more additional active ingredients are 106, 6,017,919, 6,432,984, 6,509,367, 6,645,985, and 7,169, 55 present. In another embodiment, the immunophilin inhibitor 942, which are incorporated herein by reference in their and corticosteroid are present in the composition in amounts entirety. In another embodiment, the cannabinoid receptor that, when administered together to a patient having cuta agonist is any other agonist of a cannabinoid receptor known neous inflammatory disease or disorder, inhibit or reduce in the art. Each possibility represents a separate embodiment inflammation caused therefrom. In another embodiment, the of the present invention. 60 active ingredients are present in amounts that treat the In another embodiment, the cannabinoid receptor agonist disorder. Each possibility represents a separate embodiment of methods and compositions of the present invention targets of the present invention. a type 1 (CB1) receptor. In another embodiment, the can In another embodiment of methods and compositions of nabinoid receptor agonist targets a type 2 (CB2) receptor. In the present invention, a topical composition comprises an another embodiment, the cannabinoid receptor agonist tar 65 additional active ingredient besides the immunophilin gets a WIN receptor. In another embodiment, the cannabi inhibitor and corticosteroid. In another embodiment, more noid receptor agonist targets an abnormal-cannabidiol (abn than one additional active ingredients are present. In another US 9,623,000 B2 25 26 embodiment, two additional active ingredients are present. EXAMPLES In another embodiment, more than two additional active ingredients are present. Each possibility represents a sepa Reference is now made to the following examples, which rate embodiment of the present invention. together with the above descriptions, illustrate the invention The immunophilin inhibitor of methods and compositions in a non limiting fashion. of the present invention is, in another embodiment, a non immunosuppressive immunophilin inhibitor. Non-immuno Example 1 suppressive immunophilin inhibitors are well known in the art, and include Astellas Pharmaceuticals FK 1706; Guild A study was undertaken in order to evaluate the effect of 10 topical preparations of various test items (TI) and TI com ford (now MGI) Pharmaceuticals' GPI 1046 and GPI 1485; bined with Croton oil on ear swelling. Ear thickness was 2,2'-(1,3,4-oxadiazole-2,5-diyl)bis 1-(3.3-dimethyl-1,2-di measured before croton oil induced ear swelling and 3 hours oxopentyl)-pyrrolidine (GM-284); (3R)-4-(p-Toluenesulfo post ear edema induction. The various TIs were applied nyl)-1,4-thiazane-3-carboxylic acid-L-Leucine ethyl ester, topically 1 hour prior to ear edema sensitization and 2 hours and (3R)-4-(p-Toluenesulfonyl)-(-14thiazane-3-carboxylic 15 post ear edema sensitization. The difference in ear thickness acid-L-phenylalanine benzyl ester. Additional non-immuno of the intact ear was subtracted from the ear thickness of the suppressive immunophilin inhibitors are described in Adal inflamed ear. This value is defined as “ear swelling. steinsson, H., and T. C. Bruice. 2000. Bioorg. Med. Chem. Materials and Methods 8:625-635: Gold B G. Expert Opin Investig Drugs. 2000 Test System October 9(10):2331-42. PMID: 11060810; Gold, B. G., M. Species/Strain: Mice/Balb/c Zeleny-Pooley, M. S. Wang, P. Chaturvedi, and D. M. Source: Harlan Israel, Ltd. Armistead. 1997. Exp. Neurol. 147:269-278; Guo, X. V. L. Age: Young adults, 6-7 weeks of age at study initiation. Dawson, and T. M. Dawson. 2001. Eur. J. Neurosci. Body Weight: Weight variation of animals at the time of 13:1683-1693; Sabatini, D. M., M. M. Lai, and S. H. Snyder. treatment initiation did not exceed-t15% of the mean weight. 1997. Mol. Neurobiol. 15:223-239; Sauer, H., J. M. Francis, 25 Animals Health: The health status of the animals used in this H. Jiang, G. S. Hamilton, and J. P. Steiner. 1999. Brain Res. study was examined on arrival. Only animals in good health 842:109-118; Sich, C. S. Improta, D. J. Cowley, C. Guenet, were acclimatized to laboratory conditions and were used in J. P. Merly, M.Teufel, and V. Saudek. 2000. Eur. J. Biochem. the study. 267:5342-5355; Steiner, J. P., M. A. Connolly, H. L. Valen Acclimation: 5 days. tine, G. S Hamilton, T. M. Dawson, L. Hester, and S. H. 30 Housing: During acclimation and following dosing, animals Snyder. 1997. Nat. Med. 4:412–428. Additional non-immu were housed within a limited access rodent facility in nosuppressive immunophilin inhibitors are described in U.S. conventional cages and kept in groups of maximum 10 mice, Pat. Nos. 7,153,883, 6,486,151, 6,331,537, 6,054,452, and in polypropylene cages, fitted with solid bottoms and filled 5,846,979 and U.S. Patent application numbers 2003/ with sterile wood shavings as bedding material. 0186961, 2002/0198250, and 2001/0034362, which are 35 Food and Water: Animals were provided with ad libitum—a incorporated herein by reference. commercial sterile rodent diet and free access to sterile In another embodiment, the immunophilin inhibitor is any drinking water, Supplied to each cage via polyethylene other immunophilin inhibitor known in the art. Non-limiting bottles with stainless steel sipper tubes. Feed lot analysis of examples of immunophilin inhibitors are cyclophilin, the diet batch used in the study was included in the archives FK506 binding protein (FKBP), rapamycin, and analogues 40 with the study data. thereof. Each immunophilin inhibitor represents a separate Environment: Automatically controlled environmental con embodiment of the present invention. ditions were set to maintain temperature at 20-24°C. with a It is expected that during the life of this patent many relative humidity (RH) of 30-70%, a 12:12 hour light: dark relevant corticosteroids will be developed and the scope of cycle and 10-30 air changes/hr in the study room. Tempera the term corticosteroid is intended to include all such new 45 ture and RH were monitored daily. technologies a priori. Identification: Animals were given a unique animal identi As used herein the term “about” refers to +10%. fication ear number. This number also appeared on a cage Additional objects, advantages, and novel features of the card, visible on the front of each cage. The cage card also present invention will become apparent to one ordinarily contained the study number, and all other relevant details as skilled in the art upon examination of the following 50 to treatment group and dose level. examples, which are not intended to be limiting. Addition Randomization: Animals were randomly assigned to experi ally, each of the various embodiments and aspects of the mental groups. present invention as delineated hereinabove and as claimed Termination: At the end of the study surviving animals were in the claims section below finds experimental Support in the euthanized by Isoflurane. following examples. EXAMPLES Reference is now made to the 55 Test Groups and Dosages following examples, which together with the above descrip The Experimental groups used in this study are listed in tions, illustrate the invention in a non limiting fashion. Table 1 below. TABLE 1. the 12 experimental groups comprising the study

Ear Group Group Test Volume Thickness & Gender Size Item Route Concentration (ml) Regime Measurement 1F 4 Vehicle Topical O 0.1 1 hour 24 hours 2F 4 Amitriptyline 2 mg/ml prior to prior to US 9,623,000 B2 27 28 TABLE 1-continued the 12 experimental groups comprising the study

Ear Group Group Test Volume Thickness & Gender Size Item Route Concentration (ml) Regime Measurement 3F 4 Topiramate 2 mg/m sensitization sensitization 4F 4 Moclobimide 2 mg/m and 2 hours and 3 5F 4 PEA 5 mg/m post hours post 6F 4 CBD 5 mg/m sensitization sensitization 7F 4 Hydrocortisone 600 nmolear 8F 4 Hydrocortisone + 600 nmolear + Amitriptyline 2 mg/m 9F 4 Hydrocortisone + 600 nmolear + Topiramate 2 mg/m 1OF 5 Hydrocortisone + 600 nmolear + Moclobimide 2 mg/m 11F 5 Hydrocortisone + 600 nmolear + PEA 5 mg/m 12F 5 Hydrocortisone + 600 nmolear + CBD 5 mg/m

Test Procedure Results 100 mL (50 mL each side) of freshly prepared Croton oil As is shown in Table 3 below and in FIG. 1, the ears of in acetone (2.5% V/v) solution was applied on Left ear on the Vehicle treated animals (Group 1F) swelled significantly 25 over baseline. The difference between the inflamed and Study Day 0 to induce sensitization. Prior to sensitization intact ear thickness was 0.073+0.012 mm. Treatment with animals were anesthetized. Earthickness was measured 24 the topical preparation containing CBD (Group 6F) resulted hours prior to croton oil sensitization; this measurement in a significant (72%) inhibition in ear edema compared to served as a baseline. On sensitization day, the animals were the Vehicle group (0.020+0.010 p<0.05). Treatment with the anesthetized using Ketamine and Xylazine formulation. topical preparation containing PEA (Group 5F) resulted in a Then, the animals were administered with TI topically. One 30 52% inhibition of ear swelling, which was not statistically hour later the animals were introduced to croton oil sensi significant. Treatment with the Hydrocortisone preparation tization (50 uL). Two hours post croton oil sensitization the (Group 7F) inhibited ear swelling by 40% (0.043+0.003 animals were administered with TI again. Three hours post mm) which was not statistically significant compared to the croton oil sensitization the earthickness of the animals were vehicle group. 35 Treatment with a combination of the topical preparation measured again. containing PEA and the preparation containing Hydrocorti TIs were administered twice, the first dose was 1 hour prior sone (Group 11F) resulted in 78% inhibition of ear edema to croton oil sensitization and the second time was 2 hours (0.016+0.004 p<0.01 vs. Vehicle and p<0.01 vs. Hydrocor post croton oil sensitization. All TIs were dissolved in 10% tisone). Ethanol and were applied topically on the surface of the 40 In view of the findings obtained herein it can be concluded sensitized ear. that the CBD test preparation resulted in a significant (72%) Observations and Examinations inhibition in ear edema compared to the vehicle. The PEA Determination of individual body weights of animals was test preparation showed increased (52%) but not significant made one day before sensitization phase. A single person inhibition of ear Swelling in comparison to the hydrocorti was responsible for ear thickness measurement throughout 45 sone group (40%). However, a combination of the PEA test the entire study. The ear thickness of both left and right ears preparation and the Hydrocortisone test preparation demon was measured using analogue calipers (Instrumentation 0.01 strated a significant reduction in ear edema in comparison to mm L-01) on 2 occasions: 24 hours prior to croton oil the vehicle group and the hydrocortisone group. sensitization, and 3 hours post croton oil sensitization. All data are presented in MEAN-SEM. T-test, two tailed, 50 TABLE 2 unpaired was applied to all data. P value was considered Mean Group Body weight (g significant if p <0.05. Treatment No abnormalities were observed following the treatment Groupii & Gender MEAN SEM with all test items. Animal #2 from Group 3F died post 55 anesthesia. The death of this animal was not related to the Vehicle 18.48 O.22 Group 1F treatment. Animal #4 (Group 7F) was excluded from the Amitriptyline 18.05 O41 study as redness on ear appeared prior to sensitization with Group 2F Croton oil. The mean body weight of the animals on the day Topiramate 18.78 O.25 Group 3F of study initiation was 18.48+0.15 g. Ear thickness was 60 Moclobimide 18.78 O.86 measured before Croton oil induced ear sensitization (base Group 4F line) and 3 hours post ear edema induction. Treatment with PEA 18.45 O.S8 various TI was applied topically 1 hour prior to sensitization Group SF CBD 18.68 0.44 and 2 hours post induction. The difference in ear thickness Group 6F of the intact ear was subtracted from the ear thickness 65 Hydrocortisone 18.35 O.26 measured of the inflamed ear. This value was referred to as Group 7F ear Swelling. US 9,623,000 B2 29 30 TABLE 2-continued TABLE 3-continued Mean Group Body weight (g) Mean Group Ear Swelling (mm) and % of Ear Thickness Inhibition (% Treatment 3 Hours Post % of Ear Group# & Gender MEAN SEM Treatment Baseline Croton Oil Thickness Hydrocortisone + 18.25 O.22 Group# & Gender MEAN SEM MEAN SEM Inhibition Amitriptyline Group 8F Hydrocortisone + 1820 O.45 Hydrocortisone + CBD O.212 O.OO2 O.2SO* O.OO4 59 Topiramate Group 9F 10 Group 12F Hydrocortisone + 1834 O.49 *p < 0.05 vs. Vehicle; Moclobimide Group 1 OF **p < 0.01 vs. Vehicle: Hydrocortisone + PEA 1870 O.61 #p < 0.01 vs. Hydrocortisone Group 11F Hydrocortisone + CBD 1870 O.76 15 Conclusions Group 12F The ears of the Vehicle treated animals swelled signifi cantly over baseline and the difference between the inflamed and intact earthickness was 0.073+0.012 mm. Treatment with the topical preparation containing CBD TABLE 3 resulted in a significant (72%) inhibition in ear edema Mean Group Ear Swelling (mm) and compared to the Vehicle group (0.020+0.010 p<0.05) % of Ear Thickness Inhibition (% Hydrocortisone preparation inhibited earswelling by 40% (0.043+0.003 mm). Not statistically significant 3 Hours Post % of Ear Treatment with PEA 0.5% resulted in a 52% inhibition of Treatment Baseline Croton Oil Thickness ear Swelling, which was not statistically significant 25 versus Hydrocortisone or Vehicle (0.035+0.012 mm). Groupii & Gender MEAN SEM MEAN SEM Inhibition Treatment with a combination of Hydrocortisone and the Vehicle O.231 O.OO8 O.283 O.O12 O topical preparation containing PEA (Group 11F) Group 1F resulted in 78% inhibition of ear edema (0.016+0.004 Amitriptyline O.223 O.OO6 O.248* O.OOS 62 Group 2F p<0.01 vs. Vehicle and p-0.01 vs. Hydrocortisone). Topiramate O.223 O.OO9 O.243 O.OO7 63 30 Example 2 Group 3F Moclobimide O.219 O.OO3 O.265 0.015 52 Group 4F A study was undertaken in order to evaluate the effect of PEA O.210 O.OO4 O.268 O.OO9 52 topical preparations of Hydrocortisone, Betamethasone and Group SF PEA at various concentrations (as single agents and in CBD O.225 O.OO3 O.240* O.O11 72 35 various combinations) on ear edema in mice. Earthickness Group 6F was measured before croton oil induced ear Swelling and 6 Hydrocortisone O.230 O.OO7 O.243 O.OO9 40 Group 7F hours post ear edema induction. The preparations were Hydrocortisone + O.230 O.OO8 O.258 O.O10 55 applied topically 1 hour prior to ear edema sensitization. Amitriptyline Group 8F The difference in ear thickness of the intact ear was Hydrocortisone + O.230 O.OO7 O.26S O.O10 62 40 subtracted from the ear thickness of the inflamed ear. This Topiramate Group 9F value is defined as “ear swelling. Hydrocortisone + O.218 O.OO3 O.240* O.OO7 64 Materials and Methods Moclobimide Group 1 OF Hydrocortisone + PEA O.216 O.OO2 O.23O**ii O.OO6 78 Test systems and procedures are as described above in Group 11F Example 1, the experimental groups are listed in Table 4 below. TABLE 4 the 14 experimental groups comprising the study

Ear Group Volume Thickness Group Size Test Item Route Concentration (ml) Regime Measurement

1F 10 O.S90 PEA Topical O.S9/o 25 ul 0.5 hour 24 hours per ear prior to prior to 2F 10 196 PEA Topical 190 25 ul sensitization sensitization per ear and 6 3F 10 5% PEA Topical 59% 25 ul hours per ear post 4F 10 O.S90 Topical O.S9/o 25 ul sensitization PEA + Hydrocortisone per ear EDso 5F 10 190 Topical 190 25 ul PEA + Hydrocortisone per ear EDso 6F 10 59% Topical 59% 25 ul PEA + Hydrocortisone per ear EDso US 9,623,000 B2 31 32 TABLE 4-continued the 14 experimental groups comprising the study

Ear Group Volume Thickness Group Size Test Item Route Concentration (ml) Regime Measurement 7F O O.S90 Topica O.S9/o 25 PEA + Betamethasone be ear EDso 8F O 190 Topica 190 25 PEA + Betamethasone be ear EDso 9F O 59% Topica 59% 25 PEA + Betamethasone be ear EDso 1OF O Hydrocortisone EDso Topica 55 g/25 Jul 25 per ear be ear 11F O Hydrocortisone Topica 200 g/25 Jul 25 EDnax per ear be ear 12F O Betamethasone ED50 Topica 1 g/25 Jul 25 per ear be ear 13F O Betamethasone Topica 4 g/25 Jul 25 EDnax per ear be ear 14F O Vehicle (100% Topica O 25 Ethanol) be ear

Results 25 In view of these results it can be concluded that topical Ear Thickness results are listed in Table 6 and illustrated treatment with PEA (1%) results in a significant reduction in in FIGS. 2a-b); Ear Delta results are listed in Table 7. ear edema compared to the Vehicle. In addition, this study Significant ear Swelling was measured 6 hours post sen also demonstrated that treatment with topical preparations of sitization in Vehicle treated groups (Group 14F: 0.32+0.01 Betamethasone (1 lug/25ul per ear calculated (EDs) and 4 post sensitization vs. 0.26+0.003 baseline; p<0.01). Treat 30 ment with topical preparations of Hydrocortisone at con ug/25 Jul per ear calculated (ED) lead to significant centrations of 55 g/25ul per ear—calculated (EDs) and reduction in ear edema in a dose related manner when 200 ug/25 Jul per ear calculated (ED) was not active compared to vehicle. compared to vehicle in reducing ear thickness in the current model. 35 TABLE 5 Betamethasone at concentrations of 1 Jug/25 ul per ear calculated (EDso) and 4 ug/25ul per ear calculated (ED) Mean Group Body Weight (g demonstrated significant reduction in ear edema compared Treatment and Group # Mean SEM to vehicle in a dose related manner (Betamethasone at EDs 0.30+0.008, p<0.1: Betamethasone at ED, 0.29+0.005 40 0.5% PEA(1F) 18.9 O.38 p<0.05 vs. Vehicle). % PEA(2F) 18.4 O3S 5% PEA(3F) 19.0 O3S Topical preparations of PEA at a concentration of 1% was 0.5% PEA + Hydro. EDs (4F) 18.3 O.31 significantly active in reducing ear thickness by approxi % PEA + Hydro. EDs (5F) 18.4 O.39 mately 50% (0.29+0.004, p<0.05 vs. Vehicle). No significant 5% PEA + Hydro. EDs (6F) 17.6 O.32 activity was detected for the other preparation groups in this 45 0.5% PEA + Beta. EDs (7F) 18.2 O42 current model. % PEA + Beta. EDs (8F) 18.9 O.34 5% PEA + Beta. EDs (9F) 18.4 0.27 Thus, topical treatment with PEA (1%) resulted in a Hydro. EDso (1OF) 17.3 O.33 significant reduction in ear edema compared to the Vehicle. Hydro. ED (11F) 18.5 O4O In addition, treatment with topical preparations of Betame Beta. EDso (12F) 17.5 0.44 thasone (1 lug/25ul per ear calculated (EDs) and 4 g/25ul 50 Beta. ED (13F) 17.6 O4O per ear calculated (ED) demonstrated significant reduc 00% Ethanol (veh.)(14F) 18.9 0.44 tion in ear edema compared to vehicle in a dose related al TABLE 6

Mean Left Ear Thickness

Left Ear Thickness (mm SEM Study Day -1 Study Day O Study Day -1 Study Day 0 (24 H pre (6 H post (24 H pre (6 H post Treatment and Group # sensitization) sensitization) sensitization) sensitization) 0.5% PEA(1F) O.25 O.38 O.OO O.O2 1% PEA(2F) O.26 0.29 * * O.OO O.OO 5% PEA(3F) O.26 O.32 O.OO O.OO 0.5% PEA+ Hydro. EDs (4F) O.26 O.33 O.OO O.O1 1% PEA + Hydro. EDs (5F) O.25 O.34 O.OO O.O1 US 9,623,000 B2 33 34 TABLE 6-continued

Mean Left Ear Thickness

Left Ear Thickness (mm SEM Study Day -1 Study Day O Study Day -1 Study Day 0 (24 H pre (6 H post (24 H pre (6 H post Treatment and Group # sensitization) sensitization) sensitization) sensitization) 5% PEA + Hydro. EDso 6F) O.26 O.31 O.OO O.O1 0.5% PEA + Beta. EDs (7F) O.25 O.32 O.OO O.O1 1% PEA + Beta. EDs (8F) O.25 O.36 O.OO O.O1 5% PEA + Beta. (9F) O.25 O.34 O.OO O.O1 Hydr. EDso (1OF) O.26 O.32 O.OO O.O1 Hydro. ED (11F) O.25 O.31 O.OO O.O1 Beta. EDso (12F) O.25 O.30 * O.OO O.O1 Beta ED (13F) O.26 0.29 * * O.OO O.OO 100% Ethanol (Veh.)(14F) O.26 O.32 O.OO O.O1 * p < 0.1 vs. relevant Vehicle; ** p < 0.01 vs. relevant Vehicle

TABLE 7

Mean Left Ear Delta thickness Hydrocortisone Bethametasone Treatment Group # Delta + 6 h vs baseline SEM Treatment Group # Delta + 6 h vs baseline SEM 0.5% PEA(1F) O.13S 0.017 0.5% PEA(1F) O.13S O.O17 1% PEA(2F) 0.031: 0.004 1% PEA(2F) 0.031: O.OO)4 5% PEA(3F) O.062 0.0035% PEA(3F) O.062 O.OO3 0.5% PEA + Hydro. ED50(4F) O.O68 0.012 0.5% PEA+ Betha. ED50(7F) O.O70 O.OO8 1% PEA+ Hydro. ED50(5F) O.O90 0.010 1% PEA + Betha. ED50(8F) O. 106 O.OO9 5% PEA+ Hydro. ED50(6F) O.OS1 0.013 5% PEA+ Betha. ED50(9F) O.O81 O.O13 Hydro, ED50(1OF) O.O60 0.008 Betha. ED50(12F) O.043 0.007 Hydro. EDmax(11F) O.OS4 0.008 Betha. EDmax(13F) 0.031: O.OO)4 100% Ethanol (veh.)(14F) O.062 0.010 100% Ethanol (veh.)(14F) O.062 O.O10 *p < 0.05 vs. relevant Vehicle

Conclusions No significant activity was detected for the other prepa Significant ear Swelling was measured 6 hours post sen ration groups in this current model. sitization in Vehicle treated groups (Group 14F: 0.32+0.01 post sensitization vs. 0.26+0.003 baseline: 40 Example 3 p<0.01). Treatment with topical preparations of Hydrocortisone at A study was undertaken in order to determine the effect of concentrations of 55 g/25 Jul per ear—calculated topical application of a combined preparation of Betame (ED50) and 200 g/25 ul per ear calculated (EDmax) thasone and PEA on ear edema in mice. Earthickness was were not active compared to vehicle in reducing ear 45 thickness in the current model. measured before croton oil induced ear Swelling and 6 and Betamethasone at concentrations of 1 ug/25 Jul per ear 9 hours post ear edema induction. The preparations were calculated (ED50) and 4 ug/25 ul per ear calculated applied topically 1 hour prior to ear edema sensitization. (EDmax) demonstrated significant reduction in ear The difference in ear thickness of the intact ear was edema compared to vehicle in a dose related manner 50 subtracted from the ear thickness of the inflamed ear. This (Betamethasone at ED50 0.30+0.008, p<0.1: Betame value is defined as “ear swelling. thasone at EDmax 0.29+0.005 p<0.05 vs. Vehicle). Materials and Methods Topical PEA at a concentration of 1% was significantly Test systems and procedures are as described above in active in reducing ear thickness by approximately 50% Example 1, the experimental groups are listed in Table 8 (0.29+0.004, p<0.05 vs. Vehicle). below. TABLE 8

the 4 experimental groups comprising the study

Volume Ear Group (ml) Thickness Group Size Test Item Route Concentration Per ear Regime Measurement

1F 10 Vehicle Topical O 25 Jul 0.5 hour 24 hours (100% Ethanol) prior to prior to 2F 10 19 PEA Topical 1% 25 Jul sensitization sensitization US 9,623,000 B2 35 36 TABLE 8-continued the 4 experimental groups comprising the study

Volume Ear Group (ml) Thickness Group Size Test Item Route Concentration Per ear Regime Measurement 3F 10 19 PEA- Topical 1% + 25 ul and 6 hours Betamethasone 41g25 Jul post EDnax per ear sensitization 4F 10 Betamethasone Topical 4 g/25 ul 25 ul EDnax per ear

Results sensitization (p<0.01 vs. Vehicle) but was not active at 9 Ear Thickness results are listed in Table 10 and illustrated 15 hours post sensitization. Betamethasone 4 Lig/25 l per ear in FIG. 3: Delta Thickness results are listed in Table 11. calculated (ED) (Group 4F) demonstrated significant A significant ear Swelling was measured 6 hours and 9 activityty in inhibitinglung ear swelling9. 6 hoursrs popost sensitization hours post sensitization in Vehicle treated group (Group 1F). but was not active at 9 hours post sensitization. Earthickness increased by 0.9 mm 6 hours post sensitization 20 TABLE 9 and increased by 0.13 mm 9 hours post sensitization. PEA 1% preparation (Group 2F) demonstrated significant Mean Group Body Weight (g) activity in inhibiting ear Swelling 6 hours post sensitization. (p<0.01 vs. Vehicle) but was not active in inhibiting ear Treatment and Group # Mean SEM Swelling 9 hours post sensitization. 25 A single mixture preparation of PEA (1%)+Betametha- 100% Ethanol (Veh.) (1F) 1816 O.42 Sone 4 g/25 Jul per ear calculated (ED) (Group 3F) 1% PEA (2F) 19.08 0.44 demonstrated significant activity in inhibiting ear Swelling 6 1% PEA + Beta. EDmax (3F) 18.62 O.33 hours post sensitization (p<0.001 vs. Vehicle) and main- 30 Beta. EDmax(4F) 1845 O.32 tained significant activity at 9 hours post sensitization (p<0.05 vs. Vehicle). TABLE 10

Mean Left Ear Thickness

Mean Left Ear Thickness (mm Day 0 SEM Day -1 Day O (9 h Day -1 Day O Day O Treatment and (24 h pre (6 h post post (24 h pre (6 h post (9 h post Group # sensitization) sensitization) sensitization) sensitization) sensitization) sensitization) 100% Ethanol O.25 O.34 O.38 O.OO3 O.O13 O.O17 (veh.) (1F) 1% PEA (2F) O.25 O.30** O.39 O.OO3 O.OO8 O.O24 196 O.24 0.27* * * 0.31% O.OO3 O.OO)4 O.007 PEA + Beta.EDmax (3F) Beta.EDmax (4F) O.24 0.27* * * O.33 O.OO3 O.OO6 O.O12 *p < 0.05 vs. relevant Vehicle; **p < 0.01 vs. relevant Vehicle: ***p < 0.001 vs. relevant Vehicle

Betamethasone 4 ug/25 ul per ear calculated (ED) (Group TABLE 11 4F) was active in inhibiting ear swelling 6 hours post ss sensitization (p<0.001 vs. Vehicle) but was not active at 9 Mean Left Ear Delta Thickness hours post sensitization. The single mixture preparation of PEA (1%)+Betametha- p WS+ Delta9 h WS + 6 h 9h Some 4 g/25 ul per ear calculated (ED) (Group 3F) WaS Treatment/Group # baseline baseline SEM SEM the only preparation that demonstrated significant activity at 9 hours post sensitization (p<0.05 vs. Vehicle 60 100% Ethanol (veh.)(1F) 2.98. O.133 O.O13 OO16 In view of the above results it can be concluded that a 1 % PEA(2F) 956. E. O.OO9 O.O24 preparation of PEA (1%)+Betamethasone 4 ug/25ulg4 Luper per ear Beta.1% PEA EDmax(4F) + Beta. EDmax(3F) O.O32O.O36** 0.066**O.O92ii O.OOS0.005 O.OO60.011 (Group 3F) demonstrated significant activity in inhibiting ear Swelling 6 hours post sensitization and continued to #p < 0.1, demonstrate significant activity 9 hours post sensitization 65 p < 0.05 vs. Vehicle, Topical PEA (1%) preparation (Group 2F) demonstrated **p < 0.01 vs Vehicle significant activity in inhibiting ear Swelling 6 hours post US 9,623,000 B2 37 38 Conclusions that fall within the spirit and broad scope of the appended Significant ear Swelling was measured 6 hours and 9 claims. All publications, patents and patent applications hours post sensitization in Vehicle treated group. Ear mentioned in this specification are herein incorporated in thickness increased by 0.9 mm 6 hours post sensitiza their entirety by reference into the specification, to the same tion and increased by 0.13 mm 9 hours post sensitiza extent as if each individual publication, patent or patent tion. application was specifically and individually indicated to be PEA 1% preparation demonstrated significant activity in incorporated herein by reference. In addition, citation or inhibiting ear Swelling 6 hours post sensitization. identification of any reference in this application shall not be (p<0.01 vs. Vehicle) but was not active in inhibiting ear construed as an admission that Such reference is available as Swelling 9 hours post sensitization. 10 prior art to the present invention. Betamethasone 4 ug/25ul per ear calculated was active in What is claimed is: inhibiting ear Swelling 6 hours post sensitization 1. A non-aqueous pharmaceutical composition compris (p<0.001 vs. Vehicle) but was not active at 9 hours post ing 0.5% N-palmitoylethanolamine by weight and 600 sensitization. nmole hydrocortisone and a pharmaceutically acceptable A single mixture preparation of PEA (1%)+Betametha 15 Sone 4 g/25ul per ear demonstrated significant activ carrier. ity in inhibiting ear Swelling 6 hours post sensitization 2. The pharmaceutical composition of claim 1, wherein (p<0.001 vs. Vehicle) and maintained significant activ said pharmaceutically acceptable carrier is Suitable for topi ity at 9 hours post sensitization (p<0.05 vs. Vehicle). cal administration. Combination of PEA and Betamethasone prolonged anti 3. The pharmaceutical composition of claim 1, wherein inflammatory activity said pharmaceutically acceptable carrier is Suitable for mucosal administration. It is appreciated that certain features of the invention, 4. The pharmaceutical composition of claim 1, wherein which are, for clarity, described in the context of separate said pharmaceutically acceptable carrier is Suitable for oral embodiments, may also be provided in combination in a administration. single embodiment. Conversely, various features of the 25 invention, which are, for brevity, described in the context of 5. A medical device comprising the pharmaceutical com a single embodiment, may also be provided separately or in position of claim 1. any Suitable Subcombination. 6. The medical device of claim 5, wherein said pharma Although the invention has been described in conjunction ceutical composition forms a coating of the medical device. with specific embodiments thereof, it is evident that many 30 7. The pharmaceutical composition of claim 1, formulated alternatives, modifications and variations will be apparent to as a creme, an ointment, a solution, a patch, a spray, a lotion, those skilled in the art. Accordingly, it is intended to a liniment, a varnish, or a solid preparation. embrace all such alternatives, modifications and variations k k k k k