(19) TZZ ¥__T (11) EP 2 317 857 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A01N 43/90 (2006.01) A01P 1/00 (2006.01) 28.12.2016 Bulletin 2016/52 A61K 31/4439 (2006.01) A61K 31/409 (2006.01) A61K 41/00 (2006.01) (21) Application number: 09790946.9 (86) International application number: (22) Date of filing: 29.07.2009 PCT/US2009/052128 (87) International publication number: WO 2010/014728 (04.02.2010 Gazette 2010/05) (54) USE OF TETRAKIS(N-ALKYLPYRIDINIUM)-PORPHYRIN DERIVATIVES FOR KILLING MICROBES OR PREVENTING THEIR GROWTH VERWENDUNG VON TETRAKIS(N-ALKYLPYRIDINIUM)-PORPHYRIN-DERIVATEN ZUR MIKROBENABTÖTUNG ODER ZUR VORBEUGUNG VON MIKROBENWACHSTUM UTILISATION DE DÉRIVÉS DE TÉTRAKIS (N-ALKYLPYRIDINIUM) PORPHYRINE POUR DÉTRUIRE DES MICROBES OU EMPÊCHER LEUR CROISSANCE (84) Designated Contracting States: (56) References cited: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR WO-A-01/47932 WO-A-02/02153 HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL WO-A-93/00815 WO-A-03/002077 PT RO SE SI SK SM TR WO-A-03/026646 WO-A-2004/080174 US-A1- 2003 050 296 (30) Priority: 29.07.2008 US 84403 P • KASSAB K ET AL: "PHOTOSENSITIZATION OF (43) Date of publication of application: COLPODA INFLATA CYSTS BY 11.05.2011 Bulletin 2011/19 MESO-SUBSTITUTED CATIONIC PORPHYRINS" PHOTOCHEMICAL AND PHOTOBIOLOGICAL (73) Proprietor: Frontier Scientific, Inc. SCIENCES, ROYAL SOCIETY OF CHEMISTRY, Logan, UT 84321 (US) CAMBRIDGE, GB, vol. 1, no. 8, 1 January 2002 (2002-01-01), pages 560-564, XP008066944 ISSN: (72) Inventors: 1474-905X • PRESTWICH, Glenn • BRISTOW C A ET AL: "Potential of cationic Eastbound porphyrins for photodynamic treatment of WA 98245 (US) cutaneous Leishmaniasis" PHOTODIAGNOSIS • BOMMER, Jerry, C. AND PHOTODYNAMIC THERAPY, ELSEVIER, Franklin vol. 3, no. 3, 1 September 2006 (2006-09-01), ID 83237 (US) pages 162-167, XP024978112 ISSN: 1572-1000 • TESTA, Charles [retrieved on 2006-09-01] Salt Lake City UT 84123 (US) Remarks: Thefile contains technical information submitted after (74) Representative: Pons the application was filed and not included in this Glorieta Ruben Dario 4 specification 28010 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 2 317 857 B1 Printed by Jouve, 75001 PARIS (FR) EP 2 317 857 B1 Description CROSS REFERENCE TO RELATED APPLICATIONS 5 [0001] This application claims priority upon U.S. provisional application Serial No. 61/084,403, filed July 29, 2008. BACKGROUND [0002] The use of antimicrobial agents to kill or prevent the growth of undesirable organisms has been studied exten- 10 sively. In particular, antimicrobial agents such as fungicides, antiviral, and antibacterial compounds have been examined. Although a number of antimicrobial agents are effective, they have drawbacks. For example, they can be very toxic and difficult to handle and not environmentally friendly, which limits their use. Thus, it would be desirable to have an antimi- crobial agent that can be used in a number of different applications such as, for example, aquatic applications, crop protection, and the treatment or prevention of certain diseases caused by microbes. Described herein are methods and 15 compositions that address the shortcomings of current antimicrobial agents. [0003] It has been disclosed porphyrin derivatives which are useful against gram-positive and gram-negative bacteria after photoactivation (see for example US 2003/050296 and WO 03/026646), in the dark (see for example WO 01/47932) and, also, in the dark and after irradiation (see for example Kassab, K. et al Photochemical and Photobiological Sciencies, Royal Society of Chemistry Cambridge 2002, 1, 560-564). 20 [0004] It has been also disclosed the use of porphyrin derivatives as insecticidal agents in the presence and absence of light (see for example WO 2004/080174). SUMMARY 25 [0005] Described herein are articles, compositions, and methods for killing or preventing the growth of microbes. It has been discovered that a class of porphyrins can kill or prevent the growth of microbes. The porphyrins can be used in a number of different applications where microbes grow. The advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the aspects described below. The advantages described below will be realized and attained by means of the elements and 30 combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive. BRIEF DESCRIPTION OF THE DRAWINGS 35 [0006] The accompanying drawings, which are incorporated in and constitute a part of this specification,illustrate several aspects described below. Figure 1 shows gross pictures of mice treated with different concentrations of lauryl methyl pyrifrin (LMP) and TMP. Figure 2 shows erythema scoring of lauryl methyl pyrifrin (LMP) and TMP porphyrin in an abraded area of mice. 40 Figure 3 shows erythema scoring of lauryl methyl pyrifrin (LMP) and TMP porphyrin in an intact area of mice. Figure 4 shows edema scoring of lauryl methyl pyrifrin (LMP) and TMP porphyrin in an abraded area of mice. Figure 5 shows edema scoring of lauryl methyl pyrifrin (LMP) and TMP porphyrin in an intact area of mice. DETAILED DESCRIPTION 45 [0007] In this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings: It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an" and "the" 50 include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a microbe" includes mixtures of two or more such organisms, and the like. [0008] "Optional" or "optionally" means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. 55 For example, the phrase "optionally second disinfectant" means that the second disinfectant may or may not be present. [0009] References in the specification and concluding claims to parts by weight, of a particular element or component in a composition or article, denotes the weight relationship between the element or component and any other elements or components in the composition or article for which a part by weight is expressed. Thus, in a compound containing 2 2 EP 2 317 857 B1 parts by weight of component X and 5 parts by weight component Y, X and Y are present at a weight ratio of 2:5, and are present in such ratio regardless of whether additional components are contained in the compound. [0010] A weight percent of a component, unless specifically stated to the contrary, is based on the total weight of the formulation or composition in which the component is included. 5 [0011] Variables such as R1-R4 and X used throughout the application are the same variables as previously defined unless stated to the contrary. [0012] The term "alkyl group" as used herein is a branched or unbranched saturated hydrocarbon group having 1 to 14 carbon atoms. Described herein is a topical composition comprising the porphyrin of the formula I: 10 15 20 25 30 35 1 2 4 wherein R is a C12 alkyl group, and R -R are methyl; and 40 X- is an anion; for use in the treatment or prevention of a disease caused by a microbe on a mammal; wherein the microbe is selected from a bacterium and a fungus and wherein the topical composition comprises a pharmaceutical carrier. 45 [0013] The porphyrins having the formula I can be prepared using techniques known in the art. For example, U.S. Patent No. 6,573,258 discloses techniques for preparing the porphyrins described herein, which are incorporated by 1 reference. In one aspect, R in formula I is a C 12, straight chain branched or straight alkyl group. As shown below in the Examples, the length of the alkyl group can affect the ability of the porphyrin to kill or prevent the growth of microbes. In one aspect, X is a halide, sulfate, acetate, lactate, nitrate, phosphate, carbonate, bicarbonate, or tosylate. The identity 50 of the counterion can vary depending upon how the porphyrin was synthesized. For example, if the porphyrin was alkylated with methyl iodide, the counterion would be iodide. It is also possible using techniques known in the art to exchange the counterion of the porphyrin with other counterions. For example, ion exchange resins can be used to replace one counterion for another in the porphyrin. [0014] In certain aspects, the porphyrins herein can kill up to 100% of the microbe. In other aspects, the porphyrins 55 herein can prevent the growth of the microbe. The term "prevent" as used herein includes complete cessation of growth or a reduction with the rate of growth. In general, the porphyrins described herein can kill or prevent the growth of a microbe in any environment where the organisms can exist. [0015] The porphyrins can kill or prevent the growth of a variety of different microbes. In certain aspects, the microbes 3 EP 2 317 857 B1 can be pathogenic and cause debilitating diseases to humans, livestock, and crops.