(19) TZZ_¥_T (11) EP 1 765 376 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A61K 31/198 (2006.01) A61K 8/44 (2006.01) 10.04.2013 Bulletin 2013/15 A61K 8/46 (2006.01) A61K 8/49 (2006.01) A61K 9/00 (2006.01) A61Q 19/00 (2006.01) (21) Application number: 05757465.9 (86) International application number: (22) Date of filing: 13.06.2005 PCT/US2005/020555 (87) International publication number: WO 2005/123103 (29.12.2005 Gazette 2005/52) (54) Topical compositions and use thereof for the treatment of epithelial-related conditions Topische Zusammensetzungen und Verwendung davon zur Behandlung von epithelialbedingte Leiden Compositions topiques et utilisation de celles-ci pour lutter contre des maladies epitheliales (84) Designated Contracting States: (74) Representative: ABG Patentes, S.L. AT BE BG CH CY CZ DE DK EE ES FI FR GB GR Avenida de Burgos, 16D HU IE IS IT LI LT LU MC NL PL PT RO SE SI SK TR Edificio Euromor 28036 Madrid (ES) (30) Priority: 12.06.2004 US 579093 P 14.02.2005 US 652921 P (56) References cited: WO-A-2004/087064 US-A- 5 043 268 (43) Date of publication of application: US-A- 5 202 456 US-A- 5 202 456 28.03.2007 Bulletin 2007/13 US-A- 5 705 528 (73) Proprietor: SIGNUM BIOSCIENCES, INC. • CLARKE ET AL: "Ras antagonist Monmouth Junction, NJ 08852 (US) farnesylthiosalicylic acid (FTS) reduces glomerularcellular proliferation and macrophage (72) Inventors: number in rat Thy- 1 nephritis" JOURNAL OF THE • STOCK, Jeffry B. AMERICAN SOCIETY OF NEPHROLOGY, vol. 14, Rocky Hill, NJ 08553 (US) 2003, pages 848-854, XP002518799 • GORDON, Joel • DING J ET AL: "FARNESYL-L-CYSTEINE Princeton Junction, NJ 08550 (US) ANALOGS CAN INHIBIT OR INITIATE • STOCK, Maxwell SUPEROXIDE RELEASE BY HUMAN Rocky Hill, NJ 08553 (US) NEUTROPHILS" JOURNAL OF BIOLOGICAL • STOCK, Gregory CHEMISTRY, AMERICAN SOCIETY OF Princeton, NJ 08542 (US) BIOLOCHEMICAL BIOLOGISTS, BIRMINGHAM,; US, vol. 269, no. 24, 17 June 1994 (1994-06-17), pages 16837-16844, XP001151963 ISSN: 0021-9258 Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 1 765 376 B1 Printed by Jouve, 75001 PARIS (FR) 1 EP 1 765 376 B1 2 Description they interact, or other intracellular signaling proteins, can be used to mitigate leukocyte responses and, theoreti- CROSS REFERENCE TO RELATED APPLICATIONS cally, to treat inflammatory-related conditions. (see e. g. Volker et al., Methods Enzymol., 1995,250, 216-225). [0001] This application claims the benefit of priority of 5 WO 2004/087064 describes a method for the treatment U.S. Provisional Application No. 60/579,093 filed on June of neoplasia, including cancer and other diseases and 12, 2004. This application also claims the benefit of pri- conditions in humans and mammals. More particularly, ority of U.S. Provisional Patent Application No.it provides a method for the use of prenylcysteine analogs 60/652,921 filed on February 14, 2005. The entire dis- for the treatment of neoplasia, hyperproliferative cell closures of U.S. Provisional Patent Application No.10 growth including psoriasis, restenosis following cardio- 60/597,093 and U.S. Provisional Patent Application No. vascular surgery, hyperplasia, including renal hyperpla- 60/652,921 are herein incorporated by reference. sia, chronic inflammatory diseases including rheumatoid and osteoarthritis, among others. US5043268 describes FIELD OF THE INVENTION a specific class of compounds as substrates and/or in- 15 hibitors specifically for prenyl cysteine methyltransferase [0002] The present invention relates to pharmaceuti- enzymes which catalyze the methyl transfer from S-ad- cal, cosmetic and cosmeceutical topical compositions enosylmethionine to the C-termini of proteins and pep- containing polyisoprenyl-protein inhibitor compounds tides with prenylated cysteine residus at their C-termini. and methods useful in the promotion of healthy epitheli- The compounds, as substrates, are described as being um and the treatment of epithelial-related conditions. 20 particularly suitable for use in assays for qualitatively or quantitatively detecting the farnesyl cysteine methyl- BACKGROUND OF THE INVENTION transferase enzymes. [0006] One such compound is N-acetylfar- [0003] Many skin or mucosal membrane conditions or nesyl-cysteine (AFC). AFC has been shown to inhibit disorders result from inflammation caused by, inter alia 25 membrane-associated polyisoprenoid methyl trans- bacteria, fungi, viruses, parasites, autoimmune disor- ferase and to block some neutrophil, macrophage, and ders, allergens, environmental conditions, such as ex- platelet responses in vitro. Unfortunately, AFC requires treme temperatures, wounds, hormones and/or malig- high concentrations for efficacy and is expected to result nant agents. Thus, inflammation can be associated with in generalized systemic effects and multiple side effects numerous underlying conditions ranging from dry skin to 30 since it interferes with a central cell regulation mecha- infections to cancer, as well as being symptomatic of in- nism, characteristics which would seem to preclude its flammatory disorders such as dermatitis. use in vivo. However, because such inhibitory com- [0004] Inflammation is often characterized by a strong pounds have the potential to be highly efficacious, there infiltration of leukocytes at the site of inflammation, par- is a need in the art for compositions containing these ticularly neutrophils (polymorphonuclear cells). These 35 compounds that can act as a safe and effective antidote cells promote tissue damage by releasing toxic substanc- for skin and mucosal membrane conditions. es at the vascular wall or in uninjured tissue. [0007] The invention provides a topical composition [0005] Neutrophil infiltration results from amplifying comprising: cascades of cell-cell communication involving signal transduction proteins such as G-proteins that can facili- 40 at least one polyisoprenyl-protein inhibitor com- tate intracellular regulation and intercellular communica- pound, wherein the at least one polyisoprenyl-pro- tion by interactingwith a widerange of different regulatory tein inhibitor compound comprises a compound of receptor-transducer proteins such as membrane-bound Formula 1; receptors. For these interactions to occur, many of the signal transduction proteins, including virtually all G- pro- 45 teins, must first be modified by the post- translational ad- dition of a C 15 farnesyl or C20 geranylgeranyl polyiso- prenoid moiety in thioether linkage to a cysteine residue located at or near the carboxy terminus within a so- called or its pharmaceutically acceptable salts and esters CAAX box or related cysteine- containing sequence. Car- 50 thereof, wherein boxy terminal polyisoprenoid cysteines that ultimately re- R1 is alkyl of 1 to 3 carbon atoms; 2 sult from these modifications are subject to methylester- R is --COX; wherein X is --OH, --OCH3, --NH2, 4 4 ification by a specific membrane-associated S- adenosyl- --NHR , --N(R )2 or halogen; methionine-dependent polyisoprenyl-S-cysteinyl meth- R3 is a straight or branched chain alkyl of 10 to 25 yltransferase. Compounds that can inhibit these enzy- 55 carbon atoms or a straight or branched chain alkenyl matic reactions or otherwise alter the interactions among of 10 to 25 carbon atoms; and polyisoprenylated signal transduction proteins, such as R4 is an alkyl of 1 to 25 carbon atoms; and a carrier; G-proteins and the protein regulatory targets with which 2 3 EP 1 765 376 B1 4 wherein said topical composition is in the form of a pow- 10 to 25 carbon atoms; and R 4 is an alkyl of 1 to 25 carbon der, an oil, a cream, a gel, or a bandage, and wherein atoms; and the topicalcomposition is formulatedto deliveran amount wherein the topical composition is formulated to deliver of at least one polyisoprenyl-protein inhibitor compound an amount of the at least one polyisoprenyl-protein in- that is effective to inhibit inflammation when applied top- 5 hibitor compound that is effective to inhibit inflammation ically. when applied topically. [0008] In another embodiment, provided herein is a topical composition comprising at least one polyisopre- BRIEF DESCRIPTION OF THE FIGURES nyl-protein inhibitor compound selected from the group consisting of S-farnesylcysteine, N-acetyl-S-geranyl- 10 [0012] FIG 1. Induction of edema by TPA cysteine, N-acetyl-S-farnesylcysteine ("AFC"), also re- [0013] FIG 2. AFC, alone, does not cause mouse ear ferred to as N-acetyl-S-trans, trans-famesyl-L-cysteine, edema. N-acetyl-S-geranylgeranylcysteine ("AGGC"), far-S- [0014] FIG 3. AFC inhibits TPA-induced edema nesyl-2-mercaptoethanesulfonic acid, farnesylthio-S- [0015] FIG 4. AFC treatment produces a dose depend- acetic acid, S-farnesylmercaptosuccinic acid, S-far- 15 ent inhibition of TPA-induced MPO nesylthiotriazole, S-farnesylthiosalicylic acid ("FTS"), S- [0016] FIG 5. Histology of AFC inhibition of neutrophil farnesylthiosuccinic acid, 2-chloro-5-farnesylaminoben- infiltration zoic acid, 2-famesyl-thionicotinic acid ("FTN"), 5-fluoro- [0017] FIG 6. AFC inhibits TPA-induced neutrophil in- FTS, 5-chloro-FTS, 4-chloro-FTS, S-farnesyl-methylthi- filtration osalicylic acid and combinations thereof; and a carrier; 20 [0018] FIG 7. AFC inhibition of TPA-induced MPO ac- wherein said topical composition is in the form of a pow- tivity at different application times der, an oil, a cream, a gel, or a bandage, and wherein [0019] FIG 8A. AFC does not effect TPA-induced MPO the topicalcomposition is formulatedto deliveran amount activity in the contralateral vehicle treated ear. of at least one polyisoprenyl-protein inhibitor compound [0020] FIG 8B. Dexamethasone acts to increase inhi- that is effective to inhibit inflammation when applied top- 25 bition of TPA-induced MPO activity in the contralateral ically.