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(11) EP 2 054 051 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A61K 31/192 (2006.01) A61K 31/10 (2006.01) 27.02.2013 Bulletin 2013/09 A61P 17/00 (2006.01) A61K 31/60 (2006.01) A61K 31/618 (2006.01) (21) Application number: 07788425.2 (86) International application number: (22) Date of filing: 15.08.2007 PCT/EP2007/058438

(87) International publication number: WO 2008/020025 (21.02.2008 Gazette 2008/08)

(54) USE OF 2,5-DIHYDROXYBENZENE DERIVATIVES FOR TREATING ACTINIC KERATOSIS VERWENDUNG VON 2,5-DIHYDROXYBENZOLDERIVATEN ZUR BEHANDLUNG VON AKTINISCHER KERATOSE UTILISATION DE DÉRIVÉS DE 2,5-DIHYDROXYBENZÈNE POUR LE TRAITEMENT DE LA KÉRATOSE ACTINIQUE

(84) Designated Contracting States: • SÁENZ DE TEJADA GORMAN, Iñigo AT BE BG CH CY CZ DE DK EE ES FI FR GB GR E-28036 Madrid (ES) HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE • ANGULO FRUTOS, Javier SI SK TR E-28340 Valdemoro - Madrid (ES) Designated Extension States: • VALVERDE LÓPEZ, Serafín BA HR RS E-28036 Madrid (ES) • ROMERO GARRIDO, Antonio (30) Priority: 16.08.2006 ES 200602219 E-28400 Collado Villalba - Madrid (ES) 02.07.2007 ES 200701857 • LOZANO PUERTO, Rosa María E-28036 Madrid (ES) (43) Date of publication of application: 06.05.2009 Bulletin 2009/19 (74) Representative: ABG Patentes, S.L. Avenida de Burgos, 16D (73) Proprietor: AMDERMA PHARMACEUTICALS, LLC Edificio Euromor Piscataway NJ 08854 (US) 28036 Madrid (ES)

(72) Inventors: (56) References cited: • CUEVAS SÁNCHEZ, Pedro WO-A-96/17589 WO-A-2006/029484 E-28035 Madrid (ES) US-A1- 2005 175 559 US-A1- 2007 149 618 • GIMÉNEZ GALLEGO, Guillermo US-B1- 6 281 203 E-28036 Madrid (ES)

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 2 054 051 B1

Printed by Jouve, 75001 PARIS (FR) 1 EP 2 054 051 B1 2

Description tives for removing wrinkles and stimulating collagen syn- thesis. RELATED APPLICATIONS [0009] There is a need to find alternative treatments to the current ones for skin photoaging; both from the aes- [0001] This application claims the benefit of priority of 5 thetic and therapeutic points of view, based on the use ES Application No. P200602219, filed August 16, 2006 of active principles. andof ES Application No. P200701857, filed July 2, 2007. SUMMARY OF THE INVENTION FIELD OF THE INVENTION 10 [0010] Surprisingly, the inventors have found that [0002] The present invention relates to the use of 2,5-dihydroxybenzene derivatives, the pharmaceutically 2,5-dihydroxybenzene derivatives to manufacture med- acceptable salts and solvates thereof, as well as isomers icaments useful to prevent and/or treat t pathologies as- and prodrugs thereof are useful to prevent and/or thera- sociated to skin photoaging such as actinic keratosis,. peutically treat skin aging due to exposure to ultraviolet 15 rays B (UVB), or to exposure to sun rays in general, to BACKGROUND OF THE INVENTION treat pathologies associated to said skin photoaging such as actinic keratosis. [0003] Chronic exposure to ultraviolet B rays (UVB: [0011] Therefore, a first aspect of the present invention 290-320 nm wavelength), or solar rays, in general (com- relates to the use of a compound selected from the group prising, among other wavelengths, that of UVB radiation) 20 consisting of 2,5-dihydroxybenzenesulfonic acid (Do- produces skin aging (photoaging) due to the accumula- besilate), 2-(acetyloxy)-5-hydroxybenzenesulfonic acid, tion of DNA damage and on the structural proteins of the 5-(acetyloxy)-2-hydroxybenzenesulfonic acid; 2,5-bis skin, evidencing in the form of fine wrinkles, laxity with (acetyloxy)benzenesulfonic acid, or a pharmaceutically loss of skin elasticity, elastosis, yellowish staining with acceptable salt, solvate or isomer thereof to prepare a localizad areas of melanin hyperpigmentation (solar, ac- 25 medicament for the therapeutic and/or prophylactic treat- tinic or senile lentigo). Besides, skin photoaging is asso- men of actinic keratosis. ciated with the appearance of comedoes that are more [0012] A second aspect of the present invention relates evident in the "cutis romboidalis" at the rear part of the to a compound selected from 2,5-dihydroxybenzenesul- neck. In the histological test, an epidermic atrophy and fonicacid, 2-(acetyloxy)- 5-hydroxybenzenesulfonic acid, degenerative changes in elastic fibers of the dermis may 30 5-(acetyloxy)-2-hydroxybenzenesulfonic acid, 2,5-bis be observed (Pearse AD, Gaskell SA, Marks R. J Invest (acetyloxy)benzenesulfonic acid or a pharmaceutically Dermatol1987; 88 83-87;Berton TR, MitchellDL, Fischer acceptable salt, solvate or isomer thereof, for use in the SM, Looniskar MF. J Invest Dermatol 1997;109: 340- therapeutic and/or prophylactic treatment of actinic kera- 347). Furthermore, chronic exposure to UVB rays is a tosis. risk factor for the appearance of benign lesions, such as 35 seborrheic keratosis, and premalignant lesions such as BRIEF DESCRIPTION OF THE DRAWINGS actinic keratosis (Kripke ML. Cancer Res 1994; 54: 6102- 6105). Currently, there is no effective treatment for skin [0013] photoaging (Dermatol Surg. Special Issue: Cosmeceuti- cals. Invited editors: Draelos ZD, Brody HJ. 2005). 40 Figure 1. Appearance of aged skin before treatment [0004] WO 2006/029484 discloses the use of phos- (A) and after topical treatment during three months phorilated polyphenols for treating different sings of ag- with a cream containing 5% of 2,5-dihydroxybenze- ing, such as keratoses. However, this document does nesulfonic acid (B). The arrows indicate the same not mention actinic keratosis among all the different signs area of actinic lentigo before and after treatment. of aging and fails to disclose 2,5-dihydroxybenzenesul- 45 Figure 2. The topical treatment with a cream con- phonic acid and its acetyloxy derivatives. taining 5% 2,5-dihydroxybenzenesulfonic acid [0005] US 2005/0175559 discloses the use of ide- (twice a day for 11 days) on human skin (basal con- benone and its esters for the treatment of hyperpigmen- ditions shown in Figure 2A) produces a reduction in tation. the seborrheic keratosis (big circle) as well as in the [0006] US 2007/149618, which was published after the 50 viral warts (small circle), as shown in Figure 2B. priority date of the present invention, discloses the use Figure 3. Appearance of a patient’s facial seborrheic of 2,5-dihydroxybenzenesulfonic acid compounds for the keratosis before being treated (A) and after topical treatment of skin cancer, which may be selected from treatment of the affected skin area with an emulsion actinic keratosis. containing 2.5% 2,5-dihydroxybenzenesulfonic acid [0007] US 6,281,203 discloses compositions compris- 55 for 4 weeks (B). There is a reduction of the seborrheic ing salicylic acid derivatives for the treatment of skin dis- keratosis. orders. Figure 4. Development of actinic keratosis in the [0008] WO 96/17589 discloses benzoic acid deriva- scalp of a patient before being treated (A, a), after

2 3 EP 2 054 051 B1 4 topical treatment of the affected skin area with an resents the value of the control cells, whereas the emulsion containing 2.5% 2,5-dihydroxybenze- black bar shows the value in the presence of 5A- nesulfonic acid for 2 weeks (B, b) and after this 4- 2HBS (500 mM), and the stripped bar shows the val- week-treatment (C, c). There is a reduction of the ue in the presence of 2A-SHBS (500 mM). *** indi- actinic keratosis after two weeks of treatment, and 5 cates p < 0.001 regarding control through a one- fac- a more noticeable lightening of the lesion after 4 tor analysis of variance (ANOVA) followed by a Stu- weeks of treatment. dent-Newmann-Keuls post-analysis. Figure 5. Appearance of a different lesion of actinic Figure 12. Co-crystallized potassium 5-ace- keratosis in the same patient as in Figure 4, before toxy-2-hydroxybenzenesulfonic acid with fibroblast being treated (A, a) and after topical treatment of the 10 growth factor-1. The electron density of the com- affected skin area with an emulsion containing 2.5% pound, contoured at 1 σ (panel C), enables the local- 2,5-dihydroxybenzenesulfonic acid for 4 weeks (B, ization and recognition of the compound orientation b). In this lesion, there is also a clear recovery as a regarding the protein (panels A and B), as well as result of the treatment. the confirmation that the compound maintains the Figure 6. Appearance of a lesion of actinic keratosis 15 acetoxyl group in position 2 when it binds to the pro- in the ear of a patient before being treated (A), after tein. The compound is located at a site very close to 12 days of topical treatment of the affected skin area the site that, as described, is occupied by the 2,5- with a cream containing 5% 2,5-dihydroxybenze- dihydroxybenzenesulfonic acid, which aromatic ring nesulfonic (B) and after 15 days with this treatment forms a cation-π bond with the Nε group of lysine (C). There is a reduction of the actinic keratosis after 20 132, marked in panel A as reference. Panel B shows, 12 days of treatment, and the lesion was cleared in the form of a mesh, the Van der Waals volume of after 15 days of treatment. 2-acetoxy-5-hydroxybenzenesulfonic acid, over- Figure 7. Appearance of a lesion of actinic keratosis lapped to its representation in the form of rods. In in the nose of a patient before being treated (A), and panels A and B, the protein surface is colored ac- after 30 days of topical treatment of the affected skin 25 cording to its electrostatic potential (light grey: neg- area with a cream containing 5% 2,5-dihydroxyben- ative charge; dark grey: positive charge; white: lack zenesulfonate (B). There is a reduction of the actinic of charge). keratosis after 30 days of treatment. Circle delimi- Figure 13. Co-crystallized potassium 5-ace- tates the treated area. toxy-2-hydroxybenzenesulfonic acid with fibroblast Figure 8. Inhibition of the mitogenesis induced by 30 growth factor-1. The electron density of the com- fibroblast growth factor-1 in Balb/c 3T3 fibroblast qui- pound, contoured at 1 σ (panel C), enables the local- escent cultures by calcium 2-acetoxy- 5-hydroxyben- ization and recognition of the compound orientation zenesulfonate (2A-5HBS) and potassium 2,5-dihy- regarding the protein (panels A and B) as well as the droxybenzenesulfonate (DHBS). confirmation that the compound maintains the ace- Figure 9. Inhibition of the mitogenesis induced by 35 toxyl group in position 5 when it binds to the protein. fibroblast growth factor-1 in Balb/c 3T3 fibroblast qui- The compound is located at a site very close to the escent cultures by potassium 5-acetoxy-2-hydroxy- site that, as described, is occupied by the 2,5-dihy- benzenesulfonate (5A-2HBS) and potassium 2,5- di- droxybenzenesulfonic acid, which aromatic ring hydroxybenzenesulfonate (DHBS). forms a cation-π bond with the Nε group of lysine Figure 10. Inhibition of the mitogenesis induced by 40 132, marked in panel A as reference. Panel B shows, fibroblast growth factor-1 in Balb/c 3T3 fibroblast qui- in the form of a mesh, the Van der Waals volume of escent cultures by potassium 2,5-diacetoxybenzene 2-acetoxy-5-hydroxybenzenesulfonic acid, over- sulfonate (DABS) and potassium 2,5- dihydroxyben- lapped to its representation in the form of rods. In zene sulfonate (DHBS). panels A and B, the protein surface is colored ac- Figure 11. Shows the effect of the treatment with 45 cording to its electrostatic potential (light grey: neg- potassium 5-acetoxy-2-hydroxybenzenesulfonate ative charge; dark grey: positive charge; white: lack (5-mono acetylated dobesilate; 5A-2HBS) and the of charge). potassium 2-acetoxy-5-hydroxybenzenesulfonate Figure 14. Co-crystallized 2,5-diacetoxybenze- (2-mono acetylated dobesilate; 2A-5HBS) on the nesulfonic acid with fibroblast growth actor-1. The proliferation of mouse C6 glioma cells. 5A- 2HBS and 50 electron density of the compound, contoured at 1σ 2A-5HBS were administered or not (control) after (panel C), enables the localization and recognition seeding C6 cells in 24 well plates (10 4 per well) until of the compound orientation regarding the protein they were fixed 48 hours later. Data is expressed as (panels A and B) as well as the confirmation that the the meantSEM of the percentage of absorbance at compound maintains the acetoxyl groups in posi- 595 nm obtained in control cultures, which is propor- 55 tions 2 and 5 when it binds to the protein. The com- tional to the number of cells stained with crystal vi- pound is located at a site very close to the site that, olet. Data were obtained from 3 cultures for each as described, is occupied by the 2,5-dihydroxyben- treatment and 6 control cultures. The white bar rep- zenesulfonic acid, which aromatic ring forms a cati-

3 5 EP 2 054 051 B1 6

on-π bond with the Nε group of lysine 132, marked includes local or systemic infusion techniques. in panel A as reference. Panel B represents, in the [0023] The expression "penetration enhancement" or form of a mesh, the Van der Waals volume of 2,5- "permeation enhancement" refers to the increase in the diacetoxybenzenesulfonic acid, overlapped to its permeability of the skin or mucous tissue to a pharma- representation in the form of rods. In panels A and 5 cologically active compound selected in such a way that B, the protein surface is colored according to its elec- it increases the penetration rate through the skin or mu- trostatic potential (light grey: negative charge; dark cous tissue. grey: positive charge; white: lack of charge). [0024] "Excipients" or "vehicles" refers to the vehicle materials suitable for compound administration and in- DETAILED DESCRIPTION OF THE INVENTION 10 clude any such material known in the art such as, for example, any liquid, gel, solvent, liquid diluent, solubiliz- [0014] The definitions of the terms and the chemical er, or the like, which is non-toxic and which does not show groups comprised in the formulas herein are as follows: harmful interaction with any component of the composi- tion. The term"patient" refers toanimals, preferably mam- 15 [0025] Theexpression "sustained release" refersto the mals, and more preferably humans, and includes release of an active compound and/or composition such males and females, children and adults. that the blood levels of the active compound are main- tained within a desirable therapeutic range over a period [0015] The expression "effective amount" refers to the of time. The sustained release formulation may be pre- amount of compound and/or composition effective to20 pared using any conventional known method by a skilled achieve the desired purpose. in the art in order to obtain the desired release charac- [0016] The terms "treat" or "treatment" refer to the pro- teristics. phylactic use of compounds or compositions of the [0026] "Actinic keratosis" refers to dry, squamous le- present invention to avoid the symptoms of the disease sions with gritty texture formed in the external layer of or condition, or the therapeutical use to improve an ex- 25 the skin after years of exposure to ultraviolet light, such isting condition. as solar rays. [0017] The term "prodrug" refers to compounds that [0027] "Seborrheic keratosis" refers to non-cancerous rapidly convert in vivo into pharmacologically active com- growth of the external layer of the skin. pounds. Prodrug design is generally studied in Hardma [0028] "Skin photoaging" refers to an alteration in the et al. (eds.), Goodman and Gilman’s The Pharmacolog- 30 skin caused by solar exposure. ical Basis of Therapeutics, 9th ed., pages 11-16 (1996). [0029] The term "therapeutic agent" includes any ac- A thorough study is presented in Higuchi et al., Prodrugs tive agent that can be used to treat or prevent a disease as Novel Delivery Systems, vol. 14, ASCD Symposium described herein. "Therapeutic agents" include but are Series, and in Roche (ed.), Bioreversible Carriers in Drug not limited to immunomodulatory treatments, such as a Design, American Pharmaceutical Association and Per- 35 tacrolimus ointment or a pimecrolimus cream and the gamon Press (1987). like; topical corticosteroids (cream, unguents, ointments [0018] The compounds used in the invention having or gels) or systemic corticosteroids; topical or systemic one or more asymmetric carbon atoms may exist as op- immunosupressants,such as,for example, cyclosporine, tically pure enantiomers, pure diastereomers, mixtures metrotrexate, azathioprine, and the like; phototherapy; of enantiomers, mixtures of diastereomers, racemic mix- 40 emollients such as for example, white petrolatum, eucer- tures of enantiomers, diastereomeric racemates or mix- in, urea cream, mineral oil, aluminum acetate, and the tures of diastereomeric racemates. It should be clearly like; barrier creams, sucha as, for example, zinc oxide understood that the invention contemplates and includes paste, and the like; moisturizing agents, such as, for ex- these isomers and mixtures thereof within its scope. ample, menthol, camphor, and the like; local anesthetics, [0019] The term "topical" refers to the administration 45 such as, for example, lidocaine, and the like; topical cor- of a compound by applying it on the body surface and ticoids, such as, for example, triamcinolone acetate, and includes, but is not limited to, transdermal administration the like; systemic corticoids, such as, for example, pred- and administration through the mucosa. nisone, and the like; antihistamines, such as, for exam- [0020] The term "transdermal" refers to the delivery of ple, diphenhydramine, hydroxyzine, and the like; podo- a compound that enters into the bloodstream through the 50 phylline resin, locally applied; cantharin, only combined skin. with podophylline; salicylic acid, locally applied; imiqui- [0021] The expression "through the mucosa" refers to mod;bleomycin; exfoliating agents, such as,for example, the delivery of a compound that enters into the blood- alpha hydroxy acids (glycolic acid, salicylic acid, lactic stream through the mucous tissue. acid), trichloroacetic, phenols (carbolic acid, croton oil, [0022] The term "parenteral" refers to the administra- 55 and the like; diclofenac gel; fluorouracyl, 5- bleaching tion of a compound by means of a subcutaneous, intra- agents and photoprotectors, and the like. A therapeutic venous, intramuscular, intracardiac, intradermic, intra- agent includes pharmaceutically acceptable salts there- peritoneal, intrathecal or intrasternal injection; and also of, prodrugs and pharmaceutical derivatives thereof.

4 7 EP 2 054 051 B1 8

[0030] A first aspect of the present invention relates to droxybenzenesulfonate (calcium dobesilate) and potas- theuse of a compound selectedfrom thegroup consisting sium 2,5-dihydroxybenzenesilfonate (potassium dobesi- of 2,5-dihydroxybenzenesulfonic acid (Dobesilate), 2- late). (acetyloxy)-5-hydroxybenzenesulfonic acid, 5-(acetylo- [0038] The compounds used in the invention may be xy)-2-hydroxybenzenesulfonic acid; 2,5-bis(acetyloxy) 5 synthesized by one skilled in the art using conventional benzenesulfonic acid, or a pharmaceutically acceptable and commercially available methods. The synthesis of salt, solvate or isomer thereof to prepare a medicament the compounds is disclosed in, for example, US Patent for the therapeutic and/or prophylactic treatment of ac- No 5,082,941; and "The Merck Index" 13th. edition, Mer- tinic keratosis. ck & Co., R. Railway, NJ., USA, 2001; US Patents No [0031] 2,5-diydroxybenzenic derivatives are used to 10 5,082,841, 4,814,110, 4,613,332 and 4,115,648. preprare a medicament for the cosmetic, therapeutic [0039] Compounds used in the invention also may be and/or prophylactic treatment of actinic keratosis. More in the form of solvates, particularly in the form of hydrates. particularly, the actinic keratosis is selected from the The preparation of the compounds of Formula (I), as well group consisiting of hypertrophic, atrophic, bowenoid, as the solvates thereof may be carried out by one skilled and acantholythic keratosis. 15 in the art using conventional methods and commercially [0032] In this case, when the medicament is for the available reagents. therapeutic and/or prophylactic treatment of actinic kera- [0040] The scope of the present invention encompass- tosis, 2,5-dihydroxybenzene derivatives of the invention es any salt of the compounds, especially any pharma- may be used optionally and jointly with at least one of ceutically acceptable salt of the compounds. The phrase the following therapeutic agents: imiquimod, diclofenac, 20 "pharmaceutically acceptable salts" includes metal salts glycidic acid, trichloroacetic acid, colchicine, T4 endonu- or the addition salts that may be used in pharmaceutical clease, 5-fluorouracil, isotretinoin, acitretin, cidofoir, 5- forms. For example, the pharmaceutically acceptable aminolevulinic acid, methyl aminolevulinate, hypericin, salts of the compounds provided herein may be acid ad- chemotherapeutic agent, a corticosteroid, an antibiotic, dition salts, base addition salts or metal salts and they an analgesic, an immunomodulator, an immunosuppres- 25 may be synthesized from the parenteral compounds con- sant, an anti- angiogenic (including anti- VEGF, anti-FGF, taining a base or acid residue using conventional chem- anti-EGF and anti-HGF), a leukotriene modifier, an ami- ical processes. Generally, those salts are prepared, for nosalicylate, an anesthetic, a non-steroidal anti-inflam- example, by the reaction of free base or acid forms of matory, a therapy of the solubilized interleukin receptor, these compounds with a stoichiometric amount of the a cytotoxic, inhibitors of tyrosin- kinase receptors, protein 30 appropriate base or acid in water or in an organic solvent, kinase C inhibitors, and combinations of two or more or in a mixture of both. Generally, non aqueous mediums thereof. such as ether, ethyl acetate, ethanol, isopropanol or ac- [0033] In a particular embodiment, the therapeutic etonitrile are preferred. The examples of acid addition agent is selected from the group consisting of 5-aminole- salts include addition salts of mineral acids such as, for vulinic, methyl aminolevulinate and hypericine. 35 example, hydrochloride, bromhydrate, iodide hydrate, [0034] In another particular embodiment the therapeu- sulfate, nitrate, phosphate, addition salts of organic acids tic and/or prophylactic treatment of actinic keratosis fur- such as, for example, acetate, maleate, fumarate, citrate, ther comprises photodynamic therapy, cryotherapy, cu- oxalate, succinate, tartrate, malate, mandelate, meth- rettage and surgery as a coadjuvant therapy. anesulfonate and p-toluenesulfonate. The examples of [0035] 2,5-dihydroxybenzene derivatives may be op- 40 alkali addition salts include inorganic salts such as, for tionally used combined with each other in the same or in example, ammonium salts and organic alkaline salts a different ratio. Said combinations may be in the same such as, for example, diethylamine, ethylenediamine, formulation or in formulations that would be used sequen- ethanolamine, N,N-dialkylenethanolamine, triethano- tially. lamine, glutamine and basic amino acid salts. The ex- [0036] In a preferred embodiment, the compounds45 amples of metal salts include, for example, sodium, po- used in the invention are selected from the group con- tassium, calcium, magnesium, aluminum, and lithium sisiting of: salts. [0041] In some embodiments, the invention provides calcium 2,5-dihydroxybenzenesulfonate (calcium a composition comprising an ester of dobesilate, such Dobesilate); 50 as 2-acetyloxy-5-hydroxybenzenesulfonic acid, 5- acety- potassium 2,5-dihydroxybenzenesulfonate (potas- loxy-2-hydroxybenzenesulfonic acid, or 2,5-bis-acety- sium Dobesilate); loxybenzenesulfonic acid. In some embodiments, it will magnesium 2,5-dihydroxybenzenesulfonate (mag- be desirable to formulate a composition of the invention nesium Dobesilate); with an active agent such as an ester of dobesilate, for diethylamine 2,5-dihydroxybenzenesulfonate 55 example, where the ester demonstrates greater thera- (Ethamsylate). peutic efficacythan the parent compound inthe treatment or prevention of a condition described herein. In other [0037] Particularly preferred are calcium 2,5-dihy- embodiments, the invention encompasses the use of an

5 9 EP 2 054 051 B1 10 ester of dobesilate as a prodrug, for example, to treat a [0050] In another aspect, the present invention refers condition described herein, wherein the ester is metab- to the use of 2 2,5-dihydroxybenzenesulfonic (dobesi- olized to the parent compound in a patient to achieve late) or the pharmaceutically acceptable esters or salts therapeutic efficacy in the patient. thereof to prepare a drug, medicine or composition in- [0042] The phrase "pharmaceutically acceptable" re- 5 tended for the treatment of actinic keratosis. fers to physiologically tolerable molecular entities and [0051] The duration of treatment will typically depend compositions which do not typically produce an allergic on the particular condition, its severity, the condition of or similar adverse reaction, such as gastric upset, dizzi- the patient, and the like, and will readily be determined ness, and the like, when administered to a human. Pref- by one of skill in the art. Illustrative courses of therapy erably, as used herein, the term "pharmaceutically ac- 10 include 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 ceptable" means that it is approved by a regulatory agent weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, of the Federal or a state government or listed in the U.S. 12 weeks, 3.5 months, 4 months, 4.5 months, 5 months, Pharmacopeia or other generally recognized pharmaco- 6 months, 9 months, a year, or longer as needed. poeia as suitable for use in animals, and more particu- [0052] In treating a subject suffering from a disorder larly, in humans. 15 described herein, treatment may be continued until at [0043] It would be obvious to those skilled in the art least a 10 % improvement is effected in a symptom as- thatthe scope ofthe presentinvention also encompasses sociated with the condition. In other embodiments, treat- salts that are not pharmaceutically acceptable as possi- ment is continued until the subject in need of such treat- ble media to obtain pharmaceutically acceptable salts. ment experiences an improvement of at least about 20 [0044] As used herein, the term "solvate" shall refer to 20 %, at least about 30 %, at least about 40 %, preferably any form of the active compound according to the inven- at least about 50 %, preferably at least about 60 %, more tion that exhibits another molecule (most probably, a po- preferably at least about 70 %, more preferably at least lar solvent) bound to it through a non-covalent bond. Ex- about 80 %, even more preferably 90 % or greater in a amples of solvates include hydrates and alcoholates, symptom associated with a disorder described herein. 25 preferably,C 1-C6 alcoholates,for example, methanolate. [0053] In a particular embodiment of the invention, the [0045] The pharmaceutically acceptable salts may be compounds used in the invention are administered at prepared from organic or inorganic acids or bases by least once per week. In other embodiments, said com- conventional methods through the reaction of the appro- pounds are administered at least once per day. In yet priate acid or base with the compound. other embodiments, said compounds are administered [0046] A medicament comprising a compound used in 30 twice per day. In another particular embodiment, said the invention may be presented in any suitable form for compounds are administered over a period of at least administration, for example, for systemic, transdermal, about one week. In other embodiments, said compounds oral, parenteral, buccal, nasal (e.g., by inhalation), topi- are administered over a period of at least about four cal, rectal, or intravaginal administration; therefore, a weeks. medicament of the invention may include the acceptable 35 [0054] Therapeutic amounts can be empirically deter- pharmaceutical excipients or vehicles necessary to be mined and will vary with the particular condition being formulated in the desired form of administration. In a pre- treated, the subject, the particular formulation compo- ferred embodiment, the pharmaceutical composition is nents, dosage form, and the like. administered topically. [0055] In a particular embodiment, the compounds [0047] The application of the 2,5-dihydroxybenzene 40 used in the invention are present in a pharmaceutical compounds may be made independently or, in a pre- composition in an amount of at least about 1% w/w. In ferred aspect, simultaneously with the use of equivalent other embodiments, said compounds are present in a or different mixes of other 2,5-dihydroxybenzene com- pharmaceutical composition in an amount of at least pounds (including pharmaceutically acceptable salts and about 2.5% w/w, at least about 5% w/w, at least about esters) and these compounds may be in the same for- 45 10% w/w, or at least about 15% w/w. mulation or in independent formulations that would be [0056] In one aspect of the invention, the compounds simultaneously or sequentially administered. used in the invention may be administered topically in a [0048] In one embodiment,the present invention refers formulation comprising from about 0.001 % to about 30 to the use of 2,5-dihydroxybenzensulfonic acid (dobesi- % (w/w) of the compounds. In a preferred aspect of the late) or the salts or esters thereof, to prepare a drug or 50 invention, the compounds used in the invention may be medicine intended for the therapeutic treatment of actinic administered topically in a formulation comprising from keratosis. about 0.01 % to about 20 % (w/w) of the compounds. In [0049] Similarly, as previously explained in the de- anotherpreferred aspect ofthe invention, the compounds scription, the present invention refers to the use of the used in the invention may be administered topically in a 2,5-dihydroxybenzene derivatives, the pharmaceutically 55 formulation comprising from about 0.1 % to about 15 % acceptable esters or salts thereof to prepare a drug, med- (w/w) of the compounds. In a preferred aspect of the in- icine or composition intended for the treatment of actinic vention, the compounds used in the invention may be keratosis. administered topically in a formulation comprising from

6 11 EP 2 054 051 B1 12 about 0.5 % to about 10 % (w/w) of the compounds. In III. Triglycerides, such as animal and vegetable fats anotherpreferred aspectof theinvention, thecompounds and oils. Examples include, but are not limited to, used in the invention may be administered topically in a castor oil, cod liver oil, corn oil, olive oil, almond oil, formulation comprising from about 1 % to about 5 % (w/w) palm oil, sesame oil, cotton seed oil and soybean oil. of the compounds. In another preferred aspect of the 5 IV. Acetoglyceride esters, such as acetylated mono- invention, the compounds used in the invention may be glycerides. administered topically in a formulation comprising from V. Ethoxylated glycerides, such as ethoxylated glyc- about 2.5 % to about 4 % (w/w) of the compounds. The erol monostearate. topic formulation of the compounds may be administered VI. Alkyl esters of fatty acids having 10 to 20 carbon as a single dose once a day or in multiple doses several 10 atoms. Methyl, isopropyl and butyl esters of fatty ac- times a day. In a preferred aspect of the invention, the ids are useful herein. Examples inclue, but are not topical formulation which comprises 30 %, 20 %, 15 %, limited to, hexyl laurate, isohexyl laurate, isohexyl 10 %, 5%, 2.5%, 1%, 0.5%, 0.1% or 0.001% of the com- palmitate, isopropyl palmitate, decyl oleate, isodecyl pounds, is administered four times a day. In another pre- oleate, hexadecyl stearate, decyl stearate, isopropyl ferred aspect of the invention, the topical formulation15 isostearate, diisopropyl adipate, diisohexyl adipate, which comprises about 30 %, 20 %, 15 %, 10 %, 5 %, dihexyldecyl adipate, diisopropyl sebacate, lauryl 2.5%, 1%, 0.5%, 0.1% or 0.001% of the compounds, is lactate, myristoyl lactate and cetyl lactate. administered three times a day. In another preferred as- VII. Alkenyl esters of fatty acids having 10 to 20 car- pect of the invention, the topical formulation which com- bon atoms. Examples thereof include, but are not prises about 30 %, 20 %, 15 %, 10 %, 5 %, 2.5%, 1 %, 20 limited to, oleyl myristate, oleyl stearate and oleyl 0.5 %, 0.1 % or 0.001% of the compounds is administered oleate. twice a day. In another preferred aspect of the invention, VIII. Fatty acids having 10 to 20 carbon atoms. Suit- the topical formulation which comprises about 30 %, 20 able examples include, but are not limited to, pelar- %, 15 %, 10 %, 5 %, 2.5%, 1%, 0.5%, 0.1% or 0.001 % gonic, lauric, myristic, palmitic, stearic, isostearic, of the compounds, is administered once a day. 25 hydroxystearic, oleic, linoleic, ricinoleic, arachidonic, behenic and erucic acids. Topical Compositions IX. Fatty alcohols having 10 to 20 carbon atoms. Lau- ryl, myristoyl, palmitoyl, stearyl, isostearyl, hydroxy- [0057] The product of the present invention is useful stearyl, oleyl, ricinoleyl, behenyl, erucyl and 2-octyl for topical application on the skin. The compositions com- 30 dodecanol alcohols are appropriate examples of fat- prise an effective amount of the compounds used in the ty alcohols. invention, preferably from about 0.001 to 30 %. Further- X. Fatty alcohol ethers. Ethoxylated fatty alcohols more, the composition comprises a pharmaceutical ac- having 10 to 20 carbon atoms include, but are not ceptable vehicle. The appropriate vehicles remain in the limited to, lauryl, cetyl, stearyl, isostearyl, oleyl and place of application on the skin forming a continuous film 35 cholesterol alcohols having attached thereto from 1 resistant to water immersion and perspiration. Generally, to 50 ethylene oxide groups or 1 to 50 propylene the vehicle is organic and capable of containing the for- oxide groups. mulation of the invention in a diluted or dispersed form. XI. Ether-esters, such as fatty acid esters of ethox- Lotions, creams, solutions, gels and solids are the usual ylated fatty alcohols. physical forms of the composition. 40 XII. Lanolin and derivatives. Lanolin, lanolin oil, lan- [0058] Topical application means depositing or sprea- olin wax, lanolin alcohols, lanolin fatty acids, isopro- ding the compound and the compositions over the epi- pyl lanolate, ethoxylated lanolin, ethoxylated lanolin dermic tissue (including skin and oral, gingival, nasal, alcohols, ethoxylated cholesterol, propoxylated lan- etc. tissues). olin alcohols, acetylated lanolin, acetylated lanolin 45 alcohols, lanolin alcohols linoleates, lanolin alcohols Lotions ricinoleate, acetate of lanolin alcohols ricinoleate, hydrogenolysis of lanolin, and liquid or semisolid lan- [0059] Lotions contain from about 0.001 % to about olinabsorption bases are illustrativeexamples of lan- 30% of the compounds of used in the invention, from 1% olin derived emollients. to 25% of an emollient and the appropriate amount of 50 XIII. Polyhydric alcohols and polyether derivatives. water. Examples of emollients are: Propylene glycol, dipropylene glycol, polypropylene glycol 2000 and 4000, polyoxyethylene polypropyl- I. Hydrocarbon waxes and oils such as mineral oils, ene glycols, glycerol, ethoxylated glycerol, propox- petrolatum, paraffin, ceresin, microcrystalline wax, ylated glycerol, sorbitol, ethoxylated sorbitol, hydrox- polyethylene and perhydrosqualene. 55 ypropyl sorbitol, polyethylene glycol 200-6000, II. Silicone oils such as dimethylpolysiloxanes, meth- methoxy polyethylene glycols 350, 550, 750, 2000, ylphenylpolysiloxanes and water-soluble and alco- 5000, poly(ethylene oxide) homopolymers hol-soluble glycol-silicone copolymers. (100,000-5,000,000), polyalkylene glycols and de-

7 13 EP 2 054 051 B1 14

rivatives, hexylene glycol (2-methyl-2,4-pentanedi- [0061] Some emollients previously described also ol), 1,3-butylene glycol, 1,2,6-hexanetriol, ethohex- have emulsifying properties. When a lotion contains one adiol USP (2- ethyl-1,3-hexanediol), and polyoxypro- of these emollients, an additional emulsifier is not need- pylene derivatives of trimethylolpropane are suitable ed, though it can be included in the formulation. examples. 5 [0062] The balance of the composition is water. The XIV. Polyhydric alcohol esters. Mono- and di-acyl lotions are formulated by simply admixing all of the com- esters of ethylene glycol, mono- and di-acyl esters ponents together. Preferably, the compounds of Formula of diethylene glycol, mono- and di- acyl esters of pol- I are dissolved in the emollient and the resulting mixture yethylene glycol (200-6000), mono- and di-acyl es- is added into the water. Optional components such as ters of propylene glycol, polypropylene glycol 2000 10 the emulsifier or common additives may be included in monooleate, polypropylene glycol 2000 monostear- the composition. A common additive is a thickening agent ate, ethoxylatedpropylene glycol monostearate, mo- included at a level of 1% to 30% by weight of the com- no- and di- acyl esters of glycerol, poly- acyl esters of position. Examples of suitable thickening agents are: poly glycerol, ethoxylated glycerol monostearate, Cross-linked carboxypolymethylene polymers, methyl 1,3-butylene glycol monostearate, 1,3-butylene gly- 15 cellulose, polyethylene glycols, gums and bentonite. col distearate, acyl ester of polyoxyethylene polyol, acyl esters of sorbitan, and acyl esters of polyox- Creams yethylene sorbitan are suitable examples. XV. Waxes such as beeswax, spermaceti, myristoyl [0063] The compositions of the present invention may myristate and stearyl stearate. 20 be also formulated in the form of a cream. Creams contain XVI. Beeswax derivatives, such as polyoxyethylene from 0.001% to 30% of the compounds used in the in- sorbitol beeswax. These are reaction products of vention, from 5% to 50% of an emollient and the remain- beeswax with ethoxylated sorbitol of varying ethyl- der is water. The emollients, as described above, can ene oxide content that form a mixture of ether-esters. also be used in the cream formulation. Optionally, the XVII. Vegetable waxes, including, but not limited to, 25 cream may contain an emulsifier at a level from 3% to carnauba and candelilla waxes. 50%. The previously described emulsifiers would also be XVIII.Phospholipids suchas lecithin and derivatives. adequate in this case. XIX. Sterols. Examples include, but are not limited to, cholesterol and acyl esters of cholesterol. Solutions XX. Amides, such as fatty acid amides, ethoxylated 30 acyl amides and solid fatty acid alkanolamides. [0064] The compositions of the present invention may also be formulated in the form of a solution. Solutions [0060] The lotions of the invention would further con- contain from 0.001% to 30% of the compounds used in tain from 1% to 30% of an emulsifier. The emulsifiers can the inevntion, and the adequate amount of an organic be anionic, cationic or non-ionic. Examples of non-ionic 35 solvent. Organic substances useful as the solvent or a emulsifiers include, but are not limited to, fatty alcohols part of the solvent system are as follows: propylene gly- having 10 to 20 carbon atoms, fatty alcohols having 10 col, polyethylene glycol (200-600), polypropylene glycol to 20 carbon atoms condensed with 2 to 20 moles of (425-2025), glycerine, sorbitol esters, 1,2,6-hexanetriol, ethylene oxide or propylene oxide, alkyl phenols with 6 ethanol, isopropanol, diethyl tartrate, butanediol, and to 12 carbons in the alkyl chain condensed with 2 to 20 40 mixtures thereof. Such solvent systems can also contain moles of ethylene oxide, mono- and di- acyl esters of eth- water. These compositions are applied on the skin in the ylene glycol, wherein the fatty acid contains from 10 to form of a solution, or solutions are formulated in the form 20 carbons, monoglycerides wherein the fatty acid con- of aerosol and applied on the skin as a spray. Composi- tains from 10 to 20 carbons, diethylene glycol, polyeth- tions in the form of aerosol additionally contain from 25% ylene glycols of molecular weight 200 to 6000, polypro- 45 to 80% of a suitable propellant. Examples of propellants pylene glycol of molecular weight 200 to 3000, glycerol, include, but are not limited to: chlorinated, fluorinated and sorbitol, sorbitan, polyoxyethylene sorbitol, polyoxyeth- fluorochlorinated lowmolecular weight hydrocarbons. Ni- ylene sorbitan and hydrophilic wax esters. Suitable ani- trous oxide and carbon dioxide are also used as propel- onic emulsifiers include, but are not limited to, fatty acids lant gases. Enough quantity to expel the content of the saponified (soaps) with potassium, sodium, or trieth-50 cartridge is used. anolamine, wherein the fatty acid contains from 10 to 20 carbons. Other suitable anionic emulsifiers include, but Gels are not limited to, alkali metals, ammonium or substituted ammonium with alkyl sulfates, alkyl arylsulfonates and [0065] The composition in the form of gel might be sim- alkyl ethoxy ether sulfonates having 10 to 30 carbons in 55 ply obtained by the addition of a suitable thickening agent the alkyl chain and from 1 to 50 ethylene oxide units. to the composition in the form of a solution as described Suitable cationic emulsifiers include quaternary ammo- above. Suitable thickening agents have been described nium and morpholinium and pyridinium compounds. in the chapter referring to lotions.

8 15 EP 2 054 051 B1 16

[0066] Gel formulations contain from about 0.001% to may be readily envisioned, and would depend upon the about 30% of the compounds used in the invention, 5% particular dosage form, recommended dosage, intended to 75% of a suitable organic solvent, 0.5% to 20% of a patient population, and the like. The packaging may be suitable thickening agent and the required amount of wa- in any form commonly employed for the packaging of ter. 5 pharmaceuticals, and may utilize any of a number of fea- tures such as different colors, wrapping, tamper-resistant Solids packaging, blister packs or strips, and the like. [0073] The following non-limiting examples further de- [0067] The compositions in the present invention may scribe and enable one of ordinary skill in the art to make also be formulated in solid form. Such forms have the 10 and use the present invention. shape of a bar intended for the application on the lips or other parts of the body. These compositions contain from EXAMPLES about 0.001% to about 30% of the compounds used in the invention, and from about 50% to about 98% of an Example 1: Assay of 2,5-dihydroxybenzenesulfonic emollient such as the one already described. The com- 15 acid on photoaged skin. position may be further contain from about 1% to about 20% of a suitable thickening agent, such as those already [0074] In this study, 2,5-dihydroxybenzenesulfonic ac- described, and, optionally, emulsifiers and water. id, potassium salt, formulated as 5% by weight in the form [0068] Additives usually found in topical compositions, of cream has been used, since this type of formulation such as preservatives (for example, methyl and ethyl pa- 20 is a habitual procedure for topical treatments of the skin. raben), dyes and perfumes may be included in any of the Distilled water has been used as the aqueous phase of formulations described herein. the cream. The oil phase thereof may comprise cetyl al- cohol, stearyl alcohol or vaseline. Span (sorbitan mo- Application Method nooleate) is an effective emulsifier to make the cream. 25 [0075] The following example illustrates the formula- [0069] The effective amount of the compounds used tion of an effective cream for the topical treatment of skin topically will vary according to the specific circumstances photoaging and must not be construed as a limitation of of application, the duration of exposure and similar con- the scope of the invention. siderations. Generally, the amount will vary from 0.01 [0076] Active principle: 2,5-dihydroxybenzenesulfonic microgram to 50 milligrams of the compounds used in 30 acid, potassium salt, 5% by weight. the invention, per square centimeter of the epidermis ar- Excipients: cetyl alcohol (2.5%), stearyl alcohol (2.5%), ea. The amount of topical composition (the compounds liquid vaseline (30%), white soft vaseline (20%), sorbitan used in the invention and the vehicle) applied on the af- monooleate (5%) and distilled water (q.s. to 100 g). fected area may be easily determined according to the [0077] Continuous topical treatment of human photo- amount of the compounds contained therein. 35 aged skin with this cream (twice a day for 3 months) (Fig- ure 1A) results in a reduction of the wrinkles and the Kits disappearance of actinic lentigo areas, as shown in Fig- ure 1B. Lightening of skin hyperpigmentation after the [0070] In yet other embodiments, the invention pro- application of 2,5-dihydroxybenzenesulfonic acid may vides a kit or package comprising a compound as those 40 justify the use thereof to treat melanic dyschromia with used in the invention, in packaged form, accompanied hyperpigmentation as occurs, among other pathologies, by instructions for use. The compound used in the inven- in the Mongolian spot, the nevus of Ota, the nevus of Ito, tion may be packaged in any manner suitable for admin- the nevus of Becker, ephelides (freckles), the lentigo ma- istration, so long as the packaging, when considered lign melanoma, the cafe- au-lait macules or the melasma along with the instructions for administration, indicates 45 (chloasma). the manner in which the compound is to be admininis- tered. Example2: Effect of2,5-dihydroxybenzenesulfonate [0071] For example, a kit may comprise a compound on seborrheic keratosis, actinic keratosis and viral as those used in the inventionin unit dosage form, along warts. with instructions for use. For example, such instructions 50 may indicate that administration of the compound is use- [0078] Topical treatment with the above described ful in the treatment of actinic keratosis. The compound cream (twice a day for 11 days) of human skin (basal may be packaged in any manner suitable for administra- conditions shown in Figure 2A) results in a reduction of tion. For example, when the compound is in oral dosage seborrheic keratosis (big circle) as well as of viral warts form, e.g., is in the form of a coated tablet, then the kit 55 (small circle), as shown in Figure 2B. maycomprise a sealedcontainer of coatedtablets, blister [0079] Another example of the effect of 2,5-dihydroxy- strips containing the tablets, or the like. benzenesulfonate on seborrheic keratosis is shown in [0072] Various embodiments according to the above Figure 3, wherein the seborrheic keratosis is reduced

9 17 EP 2 054 051 B1 18 due to the topical treatment on the affected area for 4 pounds were used in the form of potassium salt, except weeks, twice a day, with an emulsion containing the in the first case in which calcium salt was used. The ex- 2,5-dihydroxybenzenesulfonic acid, 2.5% by weight. periments were carried out as described in Femåndez- The emulsion containing 2.5% by weight of 2,5-dihy- Tomero C et al. J Biol Chem, 2003. droxybenzenesulfonic acid was also used to treat two 5 lesions of actinic keratosis in the same patient, as shown Example 6: Effect of monoesters of 2,5-dihydroxy- in Figures 4 and 5. After 15 days of treatment with said benzenesulfonate on the proliferation of mouse gli- emulsion (twice a day), the actinic keratosis was clearly oma C6 cells. reduced, and the effect was even more evident after 4 weeks of treatment. 10 [0087] The following example shows the efficacy of the monoesters of 2,5-dihydroxybenzenesulfonic, potassi- Example 3: Effect of an emulsion of 2.5% 2,5-dihy- um 2-acetoxy-5-hydroxybenzenesulfonate (2A-5HBS) droxybenzenesulfonate on actinic keratosis. and potassium 5-acetoxy-2-hydroxybenzenesulfonate (5A-2HBS) to reduce the proliferation capacity of glioma [0080] An emulsion containing 2.5% by weight of15 cells and supports the use of the compounds in the treat- 2,5-dihydroxybenzenesulfonic acid was used to treat two ment of gliomas. lesions of actinic keratosis in the same patient, as shown [0088] The cell line used was that of mouse glioma C6 in Figures 4 and 5. After 15 days of treatment with said cells. Once attached, the cells were treated o not treated emulsion (twice a day), the actinic keratosis was clearly (controls) with (5A-2HBS) (500 mM) or (2A-5HBS) (500 reduced, and the effect was even more evident after 4 20 mM) and they were allowed to proliferate during 48 hours. weeks of treatment. Calcium salt of 2,5-dihydroxybenze- After that, the proliferation of glioma cells was evaluated nesulfonic acid was used. by staining the fixed cells with crystal violet. The number of cells is proportional to the amount of retained dye, Example 4 : Effect of a cream of 5% 2,5-dihydroxy- which was spectrophotometrically determined by meas- benzenesulfonate on actinic keratosis. 25 uring absorbance at 595 nm after removing the dye from the cells. Both monoesters of 2,5- dihydroxybenzenesul- [0081] In this study, 2,5-dihydroxybenzenesulfonic ac- fonate, (5A-2HBS) and (2A-5HBS) caused the inhibition id, potassium salt, formulated as 5% by weight in the form of the proliferation of mouse glioma cells (Figure 11). of cream has been used, since this type of formulation is a habitual procedure for topical treatments of the skin. 30 Example 7: Analysis of the structural interaction of Distilled water has been used as the aqueous phase of the esters of 2,5-dihydroxybenzenesulfonate with the cream. The oil phase thereof may comprise cetyl al- the fibroblasts growth factor-1 (FGF-1). cohol, stearyl alcohol or vaseline. Span (sorbitan mo- nooleate) is an effective emulsifier to make the cream. [0089] As of the diffraction from complex crystals of [0082] The following example illustrates the formula- 35 FGF-1: 2-acetoxy-5-hydroxybenzenesulfonic acid, tion of an effective cream for the topical treatment of skin FGF-1: 5-acetoxy-2-hydroxybenzenesulfonic acid and photoaging and must not be construed as a limitation of FGF-1 :2,5- diacetoxybenzenesulfonic acid, the complex the scope of the invention. structures were calculated and represented. In Figures [0083] Active principle: 2,5-dihydroxybenzenesulfonic 12, 13 and 14, representing the surface of the protein acid, potassium salt, 5% by weight. 40 cloured according to its electrostatic potential (red: neg- [0084] Excipients: cetyl alcohol (2.5%), stearyl alcohol ative charge, blue: positive charge, white: areas with no (2.5%), liquid vaseline (30%), white soft vaseline (20%), charge), the interaction form of the 2- acetoxy-5-hydroxy- sorbitan monooleate (5%) and distilled water (q.s. to 100 benzenesulfonic acid, 5-acetoxy-2-hydroxybenzenesul- g). fonic acid and 2,5-diacetoxybenzenesulfonic acid, re- [0085] The mentioned formulation was topically ap- 45 spectively, with the FGF-1 may be observed. The elec- plied to actinic keratosis lesions in two different patients, tronic density of the compound, contoured at 1 σ (Figures demonstrating the efficacy of the cream containing 5% 12-14, panels C), enabled the localization and determi- 2,5-dihydroxybenzenesulfonate to treat actinic keratosis nation of the orientations of the compounds regarding (Figures 6 and 7). the protein (Figures 12-14, panels A and B), as well as 50 the confirmation that the compounds keep the acetoxyl Example 5: Inhibition of fibroblasts mitogenesis in- groups in positions 2, 5 and, 2 and 5, respectively, when duced by the fibroblast growth factor-1 (FGF-1). they bind to the protein. The compounds are located at a site very close to the site that, as described, is occupied [0086] Inhibition of FGF-1 mitogenesis was observed by the 2,5-dihydroxybenzenesulfonic acid, which aro- in quiescent cultures of Balb/c 3T3 fibroblasts by 2-ace- 55 matic ring forms a cation-π bond with the Nε group of toxy-5-hydroxybenzenesulfonate (Figure 8), 5-acetoxy- lysine 132, marked in Figures 12-14, panels A, as refer- 2-hydroxybenzenesulfonate (Figure 9) and 2,5-diace- ence. toxybenzenesulfonate (Figure 10). The evaluated com- [0090] While specific embodiments of the subject in-

10 19 EP 2 054 051 B1 20 vention have been discussed, the above specification is of the solubilized interleukin receptor, a cytotoxic, illustrative and not restrictive. One skilled in the art will inhibitors of tyrosin-kinase receptors, protein kinase appreciate that numerous changes and modifications C inhibitors, and combinations of two or more there- can be made to the invention, and that such changes and of. modifications can be made without departing from the 5 spirit and scope of the invention. The full scope of the 7. Use according to any preceding claim, wherein the invention should be determined by the claims. therapeutic and/or prophylactic treatment is associ- ated to photodynamic therapy, cryotherapy, curet- tage and surgery as a coadjuvant therapy. Claims 10 8. A compoundselected from 2,5- dihydroxybezenesul- 1. Use of a compound selected from the group consist- fonic acid, 2-(acetyloxy)-5-hydroxybenzenesulfonic ing of 2,5-dihydroxybezenesulfonic acid (Dobesi- acid, 5-(acetyloxy)-2-hydroxybenzenesulfonic acid, late), 2-(acetyloxy)-5-hydroxybenzene sulfonic acid, 2,5-bis(acetyloxy)benzenesulfonic acid or a phar- 5-(acetyloxy)-2-hydroxybenzenesulfonic acid; 2,5- 15 maceutically acceptable salt, solvate or isomer bis(acetyloxy) benzenesulfonic acid, or a pharma- thereof, for use in the therapeutic and/or prophylactic ceutically acceptable salt, solvate or isomer thereof treatment of actinic keratosis. to prepare a medicament for the therapeutic and/or prophylactic treatment of actinic keratosis 9. A compound according to claim 8, which is selected 20 from: 2-(acetyloxy)-5-hydroxybenzenesulfonic acid, 2. Use according to claim 1, wherein the compound is 5-(acetyloxy)-2-hydroxybenzenesulfonic acid, 2,5- selected from the group consisting of: bis(acetyloxy)benzenesulfonic acid, calcium 2,5-di- hydroxybenzenesulfonate (calcium Dobesilate), po- calcium 2,5-dihydroxybenzenesulfonate (calci- tassium 2,5- dihydroxybenzenesulfonate (potassium um Dobesilate); 25 Dobesilate), magnesium 2,5-dihydroxybenzenesul- potassium 2,5-dihydroxybenzenesulfonate (po- fonate (magnesium Dobesilate) and diethylamine tassium Dobesilate); 2,5-dihydroxybenzenesulfonate (Ethamsylate), for magnesium 2,5-dihydroxybenzenesulfonate use in the therapeutic and/or prophylactic treatment (magnesium Dobesilate); of actinic keratosis. diethylamine 2,5-dihydroxybenzenesulfonate 30 (Ethamsylate). Patentansprüche 3. Use according to any preceding claim, wherein ac- tinic keratosis is selected from the group consisting 1. Verwendung einer Verbindung, ausgewählt aus der of hypertrophic, atrophic, bowenoid, and acan-35 Gruppe bestehend aus 2,5-Dihydroxybenzolsulfon- tholythic keratosis. säure (Dobesilat), 2-(Acetyloxy)-5-hydroxybenzol- sulfonsäure, 5-(Acetyloxy)-2-hydroxybenzolsulfon- 4. Use according to any preceding claim wherein the säure, 2,5-Bis(acetyloxy)benzolsulfonsäure oder ei- medicament is administered topically, orally, bucca- nem pharmazeutisch annehmbaren Salz, Solvat lly, parenterally, by inhalation, rectally, intravaginally 40 oder Isomer davon zur Herstellung eines Medika- or transdermally. ments zur therapeutischen und/oder prophylakti- schen Behandlung von aktinischer Keratose. 5. Use according to any preceding claim wherein the medicament comprises at least one additional ther- 2. Verwendung gemäss Anspruch 1, wobei die Verbin- apeutic agent. 45 dungaus der Gruppeausgewählt ist, bestehendaus:

6. Use according to claim 5, wherein the at least one Calcium-2,5-dihydroxybenzolsulfonat (Calci- additional therapeutic agent is selected from the umdobesilat); group consisting of imiquimod, diclofenac, glycidic Kalium-2,5-dihydroxybenzolsulfonat (Kalium- acid, trichloroacetic acid, colchicine, T4 endonucle- 50 dobesilat); ase, 5-fluorouracil, isotretinoin, acitretin, cidofoir, 5- Magnesium-2,5-dihydroxybenzolsulfonat (Ma- aminolevulinic acid, methyl aminolevulinate, hyper- gnesiumdobesilat); icin, a chemotherapeutic agent, a corticosteroid, an Diethylamin-2,5-dihydroxybenzolsulfonat antibiotic, an analgesic, an immunomodulator, an (Ethamsylat). immunosuppressant, an anti-angiogenic (including 55 anti-VEGF, anti-FGF, anti-EGF and anti-HGF), a 3. Verwendung gemäss irgendeinem der vorherge- leukotriene modifier, an aminosalicylate, an anes- henden Ansprüche, wobei die aktinische Keratose thetic, a non-steroidal anti-inflammatory, a therapy aus der Gruppe ausgewählt ist, bestehend aus hy-

11 21 EP 2 054 051 B1 22

pertropher, atropher, bowenoider und acantholythi- lat), zur Verwendung bei der therapeutischen scher Keratose. und/oder prophylaktischen Behandlung von ak- tinischer Keratose. 4. Verwendung gemäss irgendeinem der vorherge- henden Ansprüche, wobeidas Medikament, topisch, 5 oral, bukkal, parenteral, durch Inhalation, rektal, in- Revendications travaginal oder transdermal verabreicht wird. 1. Utilisation d’un composé sélectionné dans le groupe 5. Verwendung gemäss irgendeinem der vorherge- constitué par l’acide 2,5-dihydroxybenzènesulfoni- henden Ansprüche, wobei das Medikament minde- 10 que (dobésilate), l’acide 2-(acétyloxy)-5-hydroxy- stens ein zusätzliches therapeutisches Mittel um- benzènesulfonique, l’acide 5-(acétyloxy)-2-hydro- fasst. xybenzènesulfonique ; l’acide 2,5-bis(acétyloxy) benzènesulfonique, ou d’un sel, d’un solvate ou d’un 6. Verwendung gemäss Anspruch 5, wobei das min- isomère pharmaceutiquement acceptable de celui- destens eine zusätzliche therapeutische Mittel aus 15 ci pour préparer un médicament destiné au traite- der Gruppe ausgewählt ist, bestehend aus Imiqui- ment thérapeutique et/ou prophylactique de la kéra- mod, Diclofenac, Glycidsäure, Trichloressigsäure, tose actinique. Colchicin, T4-Endonuklease, 5-Fluoruracil, Isotreti- noin, Acitretin, Cidofoir, 5-Aminolevulinsäure, Me- 2. Utilisation selon la revendication 1, dans laquelle le thylaminolevulinat, Hypericin, einem chemothera- 20 composé est sélectionné dans le groupe constitué peutischen Mittel, einem Corticosteroid, einem An- par : tibiotikum, einem Analgetikum, einem Immunmodu- lator, einem Immunsuppressivum, einem Anti-An- le 2,5-dihydroxybenzènesulfonate de calcium giogenikum (einschliesslich Anti-VEGF, Anti-FGF, (dobésilate de calcium) ; Anti-EGF und Anti-HGF), einem Leukotrienmodifi- 25 le 2,5-dihydroxybenzènesulfonate de potas- kator, einem Aminosalicylat, einem Anästhetikum, sium (dobésilate de potassium) ; einem nichtsteroidalen entzündungshemmenden le 2,5-dihydroxybenzènesulfonate de magné- Mittel, einer Therapie des solubilisierten Interleukin- sium (dobésilate de magnesium) ; Rezeptors, einem cytotoxischen Mittel, Inhibitoren le 2,5-dihydroxybenzènesulfonate de diéthyla- von Tyrosin-Kinase-Rezeptoren, Protein-Kinase C- 30 mine (étamsylate). Inhibitoren und Kombinationen von zwei oder mehreren davon. 3. Utilisation selon l’une quelconque des revendica- tions précédentes, dans laquelle la kératose actini- 7. Verwendung gemäss irgendeinem der vorherge- que est sélectionnée dans le groupe constitué par henden Ansprüche, wobei die therapeutische und/ 35 la kératose hypertrophique, atrophique, bowénoïde oder prophylaktische Behandlung mit photodynami- et acantholythique. scher Therapie, Cryotherapie, Kürettage und Chir- urgie als Coadjuvanstherapie in Verbindung steht. 4. Utilisation selon l’une quelconque des revendica- tions précédentes, dans laquelle le médicament est 8. Verbindung, ausgewählt aus 2,5-Dihydroxybenzol- 40 administré par voie topique, orale, buccale, paren- sulfonsäure, 2-(Acetyloxy)-5-hydroxybenzolsulfon- térale, par inhalation, par voie rectale, intravaginale säure, 5-(Acetyloxy)-2-hydroxybenzolsulfonsäure, ou transdermique. 2,5-Bis(acetyloxy)benzolsulfonsäure oder einem pharmazeutisch annehmbaren Salz, Solvat oder 5. Utilisation selon l’une quelconque des revendica- Isomer davon, zur Verwendung bei der therapeuti- 45 tions précédentes, dans laquelle le médicament schen und/oder prophylaktischen Behandlung von comprend au moins un agent thérapeutique supplé- aktinischer Keratose. mentaire.

9. Verbindung gemäss Anspruch 8, ausgewählt aus: 6. Utilisation selon la revendication 5, dans laquelle l’au 50 moins un agent thérapeutique supplémentaire est 2-(Acetyloxy)-5-hydroxybenzolsulfonsäure, sélectionné dans le groupe constitué par l’imiqui- 5-(Acetyloxy)-2-hydroxybenzolsulfonsäure, mod, le diclofénac, l’acide glycidique, l’acide trichlo- 2,5-Bis(acetyloxy)benzolsulfonsäure, Calcium- roacétique, la colchicine, la T4 endonucléase, le 5- 2,5-dihydroxybenzolsulfonat (Calcium Dobes- fluorouracile, l’isotrétinoïne, l’acitrétine, le cidofoir, ilat), Kalium-2,5-dihydroxybenzolsulfonat (Kali- 55 l’acide 5-aminolévulinique, l’aminolévulinate de mé- umdobesilat), Magnesium-2,5-dihydroxyben- thyle, l’hypéricine, un agent chimiothérapeutique, un zolsulfonat (Magnesiumdobesilat) und Diethyl- corticostéroïde, un antibiotique, un analgésique, un amin-2,5-dihydroxybenzolsulfonat (Ethamsy- immunomodulateur, un immunosuppresseur, un an-

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ti-angiogénique (parmi lesquels un anti-VEGF, un anti-FGF, un anti- EGF et un anti- HGF), un modifica- teur des leucotriènes, un aminosalicylate, un anes- thétique, un anti-inflammatoire non stéroïdien, une thérapie du récepteur soluble de l’interleukine, un 5 cytotoxique, les inhibiteurs des récepteurs de la ty- rosine kinase, les inhibiteurs de la protéine kinase C et des combinaisons de deux ou plus de ceux- ci.

7. Utilisation selon l’une quelconque des revendica- 10 tions précédentes, dans laquelle le traitement thé- rapeutique et/ou prophylactique est associé à une thérapie photodynamique, une cryothérapie, un cu- retage et une chirurgie en tant que thérapie co ad- juvante. 15

8. Composé sélectionné parmi l’acide 2,5-dihydroxy- benzènesulfonique, l’acide 2-(acétyloxy)-5-hy- droxybenzènesulfonique, l’acide 5-(acétyloxy)-2- hydroxybenzènesulfonique, l’acide 2,5-bis(acéty- 20 loxy)benzènesulfonique ou sel, solvate ou isomère pharmaceutiquement acceptable de celui-ci, pour une utilisation dans le traitement thérapeutique et/ou prophylactique de la kératose actinique. 25 9. Composé selon la revendication 8, qui est sélection- né parmi : l’acide 2-(acétyloxy)-5-hydroxybenzène- sulfonique, l’acide 5-(acétyloxy)-2-hydroxybenzè- nesulfonique, l’acide 2,5-bis(acétyloxy)benzènesul- fonique, le 2,5-dihydroxybenzènesulfonate de cal- 30 cium (dobésilate de calcium), le 2,5-dihydroxyben- zènesulfonate de potassium (dobésilate de potas- sium), le 2,5- dihydroxybenzènesulfonate de magné- sium (dobésilate de magnésium) et le 2,5- dihydroxy- benzènesulfonate de diéthylamine (étamsylate),35 pour une utilisation dans le traitement thérapeutique et/ou prophylactique de la kératose actinique.

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REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

Patent documents cited in the description

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• PEARSE AD ; GASKELL SA ; MARKS R. J Invest • Goodman and Gilman’s The Pharmacological Basis Dermatol, 1987, vol. 88, 83-87 [0003] of Therapeutics. 1996, 11-16 [0017] • BERTON TR ; MITCHELL DL ; FISCHER SM ; • Prodrugs as Novel Delivery Systems. HIGUCHI et al. LOONISKAR MF. J Invest Dermatol, 1997, vol. 109, Bioreversible Carriers in Drug Design. American 340-347 [0003] Pharmaceutical Association and Pergamon Press, • KRIPKE ML. Cancer Res, 1994, vol. 54, 6102-6105 1987, vol. 14 [0017] [0003] • R. RAILWAY, NJ. The Merck Index. Merck & Co, • Dermatol Surg. Special Issue: Cosmeceuticals. 2005 2001 [0038] [0003] • FEMÅNDEZ-TOMERO C et al. J Biol Chem, 2003 [0086]

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