US 20010044419A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2001/0044419 A1 VOn Borstel et al. (43) Pub. Date: Nov. 22, 2001

(54) ANTIMUTAGENIC COMPOSITIONS FOR ation of application No. 08/963,831, filed on Nov. 4, TREATMENT AND PREVENTION OF 1997. PHOTODAMAGE TO SKIN Publication Classification (75) Inventors: Reid W. von Borstel, Potomac, MD (US); Fedor Romantsev, Gaithersburg, MD (US) (51) Int. Cl." ...... A61K 48/00; A61K 31/7072; A61K 31/7076; A61K 31/708 Correspondence Address: Nixon & Vanderhye P.C. 8th Floor (52) U.S. Cl...... 514/44; 514/45; 514/50, 514/46 1100 N. Glebe Rd. Arlington, VA 22201 (US) (57) ABSTRACT (73) Assignee: Pro-Neuron, Inc. A method of improving DNA repair and reducing DNA damage and for reducing mutation frequency in Skin for the (21) Appl. No.: 09/871,973 purpose of reducing consequences of exposure to Solar or (22) Filed: Jun. 4, 2001 ultraViolet radiation is disclosed. The methods comprise administering to the skin a composition containing deoxyri Related U.S. Application Data bonucleosides in concentrations Sufficient to enhance DNA repair or reduce mutation frequency in a vehicle capable of (60) Division of application No. 09/185,084, filed on Nov. delivering effective amounts of deoxyribonucleosides to the 3, 1998, now Pat. No. 6,255,290, which is a continu necessary skin cells. US 2001/0044419 A1 Nov. 22, 2001

ANTMUTAGENC COMPOSITIONS FOR (Yarosh et al., Cancer Res. 52:4227-31, 1992). A bacterial TREATMENT AND PREVENTION OF extract has been reported to increase the rate of unscheduled PHOTODAMAGE TO SKIN DNA synthesis, which is often used as an index of DNA repair activity (Kludas and Heise, U.S. Pat. No. 4,464,362), 0001. This application is a continuation-in-part of U.S. in UV-exposed skin; however, this effect was not confirmed application Ser. No. 08/963831 filed Nov. 4, 1997, herein in a Subsequent study (Natarajan et al., Mutation Research incorporated by reference. 206:47-54, 1988). FIELD OF THE INVENTION 0006 The key issue in DNA repair, however, is not 0002 This invention relates generally to treatment and necessarily the rate of lesion excision, but the fidelity of prevention of photodamage, genetic damage, and tumori repair. Agents which accelerate the excision Step of DNA genesis in Skin and other tissues caused by exposure to Solar repair can actually exacerbate damage if the cells are inca or ultraViolet radiation or other mutagens, comprising pable of accurate repair Synthesis at a rate that matches the administration of deoxyribonucleosides or esters of deox rate of excision of damaged segments of DNA (Collins and yribonucleosides to a mammal Such as a human. These Johnson, J. Cell Physiol. 99:125-137, 1979). compounds are capable of reducing DNA damage, mutation 0007 Deoxyribonucleosides or deoxyribonucleotides frequency, and tumorigenesis when applied topically before, have been added to cells in culture with variable or divergent during, or after exposure to mutagenic radiation or chemical effects on DNA damage or mutagenesis in response to mutagens. irradiation of the cells. In Some cell types, e.g. lymphocytes, which have limited capabilities for de novo deoxyribonucle BACKGROUND OF THE INVENTION otide Synthesis, exogenous deoxynucleosides are reported to improve cell Survival after exposure to UV radiation (Yew 0003 Exposure of skin to ultraviolet (or ionizing) radia and Johnson, Biochim. Biophys. Acta, 562:240-251, 1979; tion damageS DNA, which if unrepaired or improperly Green et al., Mutation Research, 350:239-246, 1996) or repaired, can lead to carcinogenesis as well as contribute to ionizing radiation (Petrovic et al., Int. J. Radiat. Biol., acceleration of the aging process. DNA damage and conse 18:243, 1970); no significant improvement in survival was quent genomic instability are defining characteristics of both seen after addition of deoxyribonucleosides to UV-irradiated carcinogenesis and biological aging. Patients with defective normal human fibroblasts (Green et al., Mutation Research, DNA repair capabilities in diseases like Xeroderma pigmen 350:239-246, 1996). A crucial point is that increasing cell tosa display premature Skin aging and a very high incidence Survival after genomic damage caused by UV radiation or of skin cancers (Robbins and Moshell, J. Inv. Dermatol, other mutagens is not necessarily desirable. The process of 73:102-107, 1979) on Sun-exposed areas of the skin. Phar programmed cell death, or apoptosis, is integrated with macological intervention in damage to skin due to Solar or cellular mechanisms for detecting DNA damage. Thus, ultraViolet radiation has heretofore been largely restricted to genomic damage which by itself is not Sufficient to cause agents like SunScreens or free-radical Scavengers intended to cell death, can trigger apoptosis, an active cellular Suicide prevent damage, or agents like retinoic acid or glycolic acid process, So that the DNA damage in the cell is not perpetu which are intended to remodel the Surface of radiation ated in Subsequent cell generations, with tumorigenesis as a damaged skin without necessarily addressing the most fun possible outcome as genomic damage accumulates. The damental mechanisms of cell or tissue damage and repair at mechanisms for detecting genomic damage and inducing the level of genomic integrity. apoptosis involve cell-cycle regulating proteins Such as the 0004. In practice, preventive measures like sunscreen use tumor-Suppressor protein p53. Therefore, agents which pro are not completely effective, and exposure to Sunlight is not mote cell Survival (e.g. by inhibiting apoptosis) after irra always anticipated. The incidence of skin cancers in the diation are not necessarily anticarcinogenic, and may actu United States approaches 1,000,000 cases per year. There ally enhance mutation frequency and risk of malignant fore, there exists a need for a therapeutic agent which will transformation by permitting Survival of damaged cells that reduce the risk of development of skin cancer or other would otherwise be eliminated by apoptosis. A significant consequences of skin photodamage even when applied after illustration of this principle is the demonstration that embry exposure to Sunlight has already occurred. Sunscreens and onic p53 knockout mice exposed to ionizing radiation in agents which induce or improve tanning are not useful in utero have a higher survival rate (live birth) than wild-type Such situations, Since they are only useful if applied prior to controls, but also have a much higher frequency of congeni exposure to UV radiation. Moreover, there are situations tal defects (Norimura et al., Nature Medicine, 2:577-580). wherein SunScreens and even endogenous melanin can actu 0008. In studies where the effect of exogenous deoxyri ally enhance UV-induced DNA damage through photody bonucleosides on mutation frequency in UV-irradiated cells namic Sensitization. has been explicitly Studied, variable results have been 0005 There have been several attempts to improve or obtained. Bianchi and Celotti (Mutation Research 146:277 accelerate DNA repair and to reduce the consequences of 284, 1985) reported that thymidine or deoxycytidine at high DNA damage in skin cells after damage has already concentrations increased the mutation frequency in UV occurred. The first major step in DNA repair is detection and irradiated V79 Chinese hamster cells; no reduction in muta excision of damaged portions of DNA. The viral enzyme T4 tion frequency was observed at any concentrations of added endonuclease V can accomplish this step with Some forms of nucleosides. Musk et al. (Mutation Research 227:25-30, DNA damage. T4 endonuclease V, when packaged in epi 1989) reported that a mixture of deoxyribonucleosides dermis-penetrating liposomes, has been shown to accelerate which included exceSS deoxycytidine relative to the other the rate of excision of pyrimidine dimers, the most common nucleosides, reduced the mutation frequency in response to form of photolesion, in photodamaged skin of mice in vivo UV-C (254 nm) irradiation in MM96L melanoma cells, a US 2001/0044419 A1 Nov. 22, 2001

cell line with a known constitutive excess of purine deox and congeners adenosine, cyclic-AMP, uridine, cytidine yribonucleotides. In the same Study, exogenous deoxyribo increased the period of latency for extravasation of Systemi nucleosides had no effect on mutation frequency in another cally administered Evan's Blue dye in the skin in the first neoplastic cell line, human HeLa cells, after exposure to few hours after UV exposure, indicating a reduction in acute UV-C radiation. It is important to note that UV-C radiation UV-induced edema. In this system, DNA administered after is not a component of Solar radiation at the Surface of the irradiation had no effect on extravasation of dye. The author earth, since it is blocked effectively by the atmosphere also explicitly States that nucleobases incorporated into (Pathak, 1974, in Sunlight and Man, ed. by T. B. Fitzpatrick, ointments do not penetrate the horny layer (the Stratum University of Tokyo Press, Tokyo, Japan, p. 815). The effect corneum, the Outer layer of enucleated keratinocytes com of deoxyribonucleosides on mutation frequency in cells prising the main moisture barrier of skin) of human epider exposed to Solar radiation or UV radiation at wavelengths S. that are present in Solar radiation was not tested, and the authors explicitly conclude their discussion with the State OBJECTS OF THE INVENTION ment “... the lowermutation frequency in Sun-irradiated compared with UVC-irradiated MM96L cells suggests that 0013. It is an object of the invention to provide compo Sunlight either does not perturb the deoxynucleoside pools Sitions and methods for reducing mutation frequency, or it induces a cellular response that is insensitive to nucleo photaging, and tumorigenesis in Skin, thereby attenuating Side levels.” In addition to agents which inhibit apoptosis or consequences of exposure to Solar and ultraViolet radiation improve DNA repair sufficiently to permit cell survival but and to other mutagens including endogenous oxidants. not necessarily for correction of potentially tumorigenic 0014. It is an object of the invention to provide a com mutations, growth factors in general (including those that position that enhances DNA repair and prevents conse are involved in normal wound healing responses like TGF-B quences of mutagenic radiation even when administered or PDGF) act as tumor promoters. after damage or exposure to radiation or other mutagens has 0009 U.S. Pat. No. 5,246,708 discloses the methods and already occurred. compositions involving the use of mixtures of deoxyribo 0015. It is a primary object of this invention to provide nucleosides for promotion of the healing of wounds, ulcers, compositions and methods for effectively preventing or and burns, including those caused by ultraViolet or Solar treating consequences of exposure of the skin to Solar and radiation. ultraViolet radiation and other environmental mutagens. 0.010 Acyl derivatives of deoxyribonucleosides have been taught as delivery molecules for promoting entry of 0016. It is a further object of the invention to provide deoxyribonucleosides into the skin, as disclosed in U.S. compositions and methods for improving the activity of patent application Ser. No. 466,379. It is disclosed that acyl chemical Sunscreens. derivatives of deoxyribonucleosides can improve cellular 0017. It is a further object of the invention to provide repair and cell Survival after damage to skin caused by compositions and methods for reducing deleterious effects radiation. of Sunscreens and other compounds, exogenous and endog 0.011 Oligodeoxyribonucleotides have been proposed as enous, which act as photosensitizing or photodynamic melanogenic Stimuli, based on the idea that DNA damage, or enhancers of UV-induced damage to skin. excision products of DNA damage, might be cellular signals 0018. It is a further object of the invention to provide for increasing melanin production in the skin to help protect compositions and methods for reducing Some consequences against Subsequent damage. Gilchrest et al. (U.S. Pat. No. of inflammatory skin and mucosal conditions, including 5,470,577; WO Application Ser. No. 95/01773) proposed pSoriasis, dermatitis and inflammatory bowel disease. that exogenous DNA photodamage products may stimulate melanogenesis without actual damage to cellular DNA as a SUMMARY OF THE INVENTION necessary intermediate Step. The Stated intention was to mimic the presence of cyclobutane pyrimidine dimerS or 0019. The subject invention involves methods and com other DNA photodamage products in order to provide the positions for improving DNA repair (or genomic fidelity) cell with false DNA damage Signals that might trigger and reducing photodamage in Skin exposed to ultraViolet induction of melanogenesis in the absence of actual DNA radiation, ionizing radiation, or chemical mutagens by topi damage. Treatment of melanoma cells in vitro and guinea cal administration of compositions containing effective pig skin in Vivo with thymidine dinucleotide resulted in amounts of a Source of deoxyribonucleosides. The compo increases in melanin production. The authors Stated that they Sitions are capable of delivering deoxyribonucleosides to the believed that DNA fragments entered the cells, and even necessary target cells. Such compositions are applied to the their nuclei, intact. They proposed that Sunless tanning skin before, during or after exposure to Solar, ultraViolet, or accomplished over a period of weeks by topical adminis ionizing radiation, or chemical mutagens including but not tration of oligodeoxyribonucleotides, especially thymidine limited to endogenous or exogenous Sources of free radicals dinucleotide, could protect Skin by inducing melanin Syn or nitric oxide. Treatment with compositions of the inven thesis, with consequent reduction of passage of UV radiation tion improves the net fidelity of DNA repair and thereby into and through the skin. reduces mutation frequency and the risk of tumorigenesis in response to Solar or ultraViolet radiation or other mutagens. 0012 Wiskemann (1974; in Sunlight and Man, ed. by T. B. Fizpatrick, University of Tokyo Press, Tokyo, Japan, p. 0020. The invention provides methods and compositions 51) reported that Systemic (intraperitoneal) administration for delivering deoxyribonucleosides to Skin cells in concen the deoxyribonucleoside thymidine or the ribonucleosides trations Sufficient to Support and improve repair of damaged US 2001/0044419 A1 Nov. 22, 2001

DNA and to reduce deleterious consequences of exposure of verted to deoxyribonucleosides by endogenous enzymes, skin to radiation or chemical mutagens. especially esterases. Examples include acyl derivatives of deoxyribonucleosides (carboxylic acid esters), deoxyribo 0021. In addition to prevention of consequences of expo nucleotides (phosphate esters), or oligodeoxyribonucle Sure to Sunlight, compounds and compositions of the inven otides (phosphate diesters). Since esterase activity (involv tion are useful for treating skin lesions caused by Sunlight ing various enzymes capable of cleaving carboxylic acid like actinic keratoses or Solar lentigenes. esters and phosphate esters) is ubiquitous in mammalian 0022. The deoxyribonucleosides are administered either tissues including skin, these esters of deoxyribonucleosides as free deoxyribonucleosides, or as derivatives thereof are converted to deoxyribonucleosides when applied to skin. which are converted to deoxyribonucleosides after applica Similarly, a “source of at least one ribonucleoside” refers to tion to the skin. Such derivatives include deoxyribonucle a ribonucleoside or ribonucleoside ester, including a ribo otides, oligonucleotides, DNA itself, and acyl derivatives of nucleotide, an oligoribonucleotide, or an acyl derivative of deoxyribonucleosides or other derivatives of deoxyribo a ribonucleoside. nucleosides which are converted to free deoxyribonucleo 0.031) The term “ester of a deoxyribonucleoside” (or Sides by endogenous enzymes. deoxyribonucleoside ester) refers to either an acyl derivative 0023 Methods and compositions of the invention also of deoxyribonucleosides as described above or to a phos improve activity and reduce Side effects of other agents used phate ester of a deoxyribonucleoside (or deoxyribonucleo on Skin for prophylactic, therapeutic, or cosmetic purposes, sides), e.g. deoxyribonucleotides, oligodeoxyribonucle including but not limited to Sunscreens, retinoids, alpha otides, or polydeoxyribonucleotides. hydroxy acids, methylxanthines, and DNA repair enzymes. 0032. The term “photosensitization” in the context of the 0024. The invention also relates to compositions and Subject invention refers to the process whereby light-absorb methods for reducing deleterious consequences (e.g. cellular ing (UV or visible light) molecules directly transfer the damage, especially to DNA, which can result in increased energy of an excited State, generally a triplet State, to a target likelihood of mutations or other potentially carcinogenic molecule, resulting in damage to DNA and other cellular damage to the genome) of endogenous and exogenous structures. The target molecule can be DNA itself or another photochemically-active compounds or chromophores which target which results in damage to DNA, e.g. membranes act as photoSensitizers or photodynamic enhancers of DNA components of lySOSomes, which contain deoxyribonu damage caused by Solar or ultraViolet radiation. clease. 0.025 The invention, as well as other objects, features and 0033. The term “photodynamic sensitization” herein advantages thereof will be understood more clearly and fully refers to the proceSS whereby UV-absorbing molecules gen from the following detailed description, when read with erate free radical Species or other diffusible reactive inter reference to the accompanying results of the experiments mediates as a result of excitation by UV or visible radiation. discussed in the examples below. 0034. The term “Sunscreen agents” refers to a UV-ab Sorbing chemicals that are intended to be used in SunScreen DETAILED DESCRIPTION OF THE products as active ingredients for reducing exposure of the INVENTION skin to the UV component of Solar radiation. Examples of Sunscreen agents currently used as Such in commercial 0.026 DNA damage and repair is involved in the devel products include avobenzone (t-butyl dimethoxydibenzoyl opment of Skin cancer and photoaging. The Subject inven methane), oxybenzone (benzophenone-3), dioxybenzone tion provides compounds which Successfully improve the (benzophenone-8), Sulisobenzone (benzophenone-4; 2-hy net fidelity of DNA repair. The subject invention will have droxy-4-methoxybenzophenone-5-Sulfonic acid), octoc important consequences in health care and will improve the rylene (2-ethylhexyl-2-cyano-3,3-diphenylacrylate), octyl cosmetic appearance of skin. methoxycinnamate (2-ethylhexyl p-methoxycinnamate), 0027 A. Definitions octyl salicylate (2-ethylhexylsalicylate), homosalate (homo 0028. The term “deoxyribonucleoside” refers to any one menthyl Salicylate), trolamine Salicylate (triethanolamine of the four principle nucleoside constituents of DNA: deoxy Salicylate), phenylbenzimidazole Sulfonic acid, PABA (para adenosine, deoxycytidine, deoxyguanosine, and thymidine. aminobenzoic acid), roXadimate (ethyl 4-bis hydroxypropyl The term “ribonucleoside” refers to any one of the four aminobenzoate), lisadimate (glyceryl PABA), Padimate O major nucleoside constituents of RNA:adenosine, cytidine, (octyldimethyl PABA), menthyl anthranilate, and Parsol guanosine, and uridine. 1789 (butyl methoxydibenzoylmethane). 0035. The term “energy scavenger” refers to a compound 0029. The term “acyl derivative of a deoxyribonucleo which absorbs energy from the excited States of SunScreen Side” refers to deoxyribonucleosides bearing acyl Substitu agents or endogenous photoSensitizing or photodynamic ents derived from carboxylic acids (which modify the phar enhancers of DNA damage, reducing damage to target macokinetics and bioavailability of the free biological molecules like DNA. In this context, energy deoxyribonucleosides), as disclosed in U.S. patent applica Scavengers should be nontoxic at effective concentrations tion Ser. No. 466,379, hereby incorporated by reference in and should yield a net reduction in damage to DNA when its entirety. present during UV irradiation in the presence of a photo 0030 The term “source of at least one deoxyribonucleo Sensitizer or photodynamic enhancer of DNA damage. In the side' or “deoxyribonucleoside source” in the context of the context of this invention, "energy Scavengers' refer particu Subject invention refers to deoxyribonucleosides themselves larly to compounds with lowest triplet States with energies or derivatives of deoxyribonucleosides which can be con less than or equal to those of the nucleobase constituents of US 2001/0044419 A1 Nov. 22, 2001 genomic DNA. The primary quality of energy Scavengers of protecting the skin against mutagens. The composition com the invention is that, by virtue of photochemical energy State prises 1) an effective amount of a Source of one or more properties, or mass action (concentration), they prevent deoxyribonucleosides, and optionally 2) an effective amount damage to genomic DNA that would otherwise occur due to of a pharmaceutically acceptable topical carrier capable of energy transfer for excited chromophores, whether endog delivering the deoxyribonucleosides or their precursors to enous (e.g. melanin) or exogenous (e.g. Sunscreens). Energy appropriate target cells in the skin under in Vivo conditions. Scavengers with photochemical properties similar to those of 0045 While individual deoxyribonucleosides, especially Structural constituents of DNA are advantageous, and deoxycytidine (see Example 7) have Some activity in attenu include Sources of at least one deoxyribonculeoside or ating UV-induced tumorigenesis, two or more deoxyribo ribonucleoside, like a deoxyribonucleoside, an acyl deox nucleosides, or preferably all four, are typically included in yribonucleoside, a deoxyribonucleotide, an oligodeoxyribo a formulation of the invention. Encompassed by the inven nucleotide, a ribonucleoside, a ribonucleotide, an oligoribo tion are compositions containing deoxyadenosine, deoxycy nucleotide, and an acyl ribonucleoside. tidine, deoxyguanosine, or thymidine, either as Single 0.036 The term “deleterious consequences” as used agents, or in all possible combinations of two, three, or all herein refers to cellular damage in a mammal caused by a four of these compounds. Compositions containing deoxy mutagen, especially damage to the genome, resulting in an cytidine are particularly advantageous. The concentrations increased chance of developing skin cancer or other skin of individual deoxyribonucleosides in compositions encom lesions like Solar lentigines, actinic keratoses, or other signs passed by the invention, whether present individually or in of photoaging like Skin wrinkles or "age spots”. Mutagens combination with other deoxyribonucleosides or deoxyribo capable of causing Such deleterious consequences include nucleoside precursors (Such as deoxyribonucleoside esters), Solar radiation, ultraViolet radiation, ionizing radiation, free and normalized to the amount of free nucleoside or nucleo radicals (whether produced as a result of irradiation of a Side moiety in the case of deoxyribonucleoside phosphates photochemically active chromophore or from Some other or oligonucleotides or prodrugs like acylated deoxyribo Source, including normal metabolic processes), nitric oxide, nucleoside derivatives, range from 0.1 to 10 mg/ml, advan and environmental mutagens. tageously 1 to 5 mg/ml. 0037 B. Compounds of the Invention 0046. In the case of deoxyribose or acyl derivatives of 0.038. The compounds of the invention are primarily the deoxyribose, appropriate concentrations in a composition of major deoxyribonucleoside constituents of DNA: deoxyad the invention range from 0.1 to 100 millimolar, advanta enosine, deoxycytidine, deoxyguanosine, and thymidine. geously 10 to 50 millimolar. The invention also includes the use of effective amounts of 0047. In order to permit access of the deoxyribonucleo precursors of these deoxyribonucleosides, e.g. oligodeox Sides and related compounds of the invention to deeper yribonucleotides, DNA, deoxyribonucleotides and acyl lying skin cells, vehicles which improve their penetration derivatives of deoxyribonucleosides, and, particularly for through the outer layers of the Skin, e.g. the Stratum cor minimization of effects of photodynamic Sensitizers and neum, are useful. Vehicle constituents which improve the photoSensitizing agents on DNA, ribonucleosides and their penetration of compounds of the invention into the skin congeners, e.g. oligoribonucleotides, ribonucleotides, and include but are not limited to: ethanol, isopropanol, dieth acyl derivatives of ribonucleosides. ylene glycol etherS Such as diethylene glycol monoethyl ether, aZone (1-dodecylazacycloheptan-2-one), oleic acid, 0.039 2-Deoxyribose and acyl derivatives of 2-deoxyri linoleic acid, propylene glycol, hypertonic concentrations of bose are also useful compounds of the invention. They are glycerol, lactic acid, glycolic acid, citric acid, and malic particularly advantageous for use in treatment of existing acid. Sunlight-induced Skin lesions like actinic keratoses. 0048. Appropriate concentrations of diethylene glycol 0040. While not wishing to be bound by a theory, it is monoehtyl ether in compositions of the invention range from believed that the active agents of the invention that pass into 2 to 20 percent, advantageously from 5 to 15 percent, on a cells are the deoxyribonucleosides or acyl derivatives of weight/weight basis. deoxyribonucleosides, Since the anionic phosphate moiety on deoxyribonucleotides or oligodeoxyribonucleotides 0049. In addition to promoting absorption of agents into impedes passage acroSS cell membranes. Phosphorylated the skin, use of topical alpha-hydroxy acids (AHA), e.g. deoxyribonucleoside precursors are converted to free deox lactic acid and glycolic acid, can affect the ability of the skin yribonucleosides by enzymatic and nonenzymatic degrada to reduce the penetration of ultraViolet light into the Vulner tion before or after application to the Skin, prior to their entry able basal layers of the epidermis. Thus, there is also an into cells. increased need for agents which reduce the consequences of exposure to Solar or ultraViolet radiation in people using 0041. The deoxyribonucleosides are produced by any of AHA's, e.g. for promoting exfoliation of epidermal cells. several methods. They are produced by degradation of DNA Since penetration of UV-absorbing Sunscreen agents into the from biological Sources, e.g. fish Sperm, by chemical Syn skin is undesirable because of possible photosensitization thesis, or by fermentation technology. and photodynamic enhancement of UV-induced damage to 0.042 Also encompassed by the invention are pharma cells, the compounds of the invention are uniquely Suitable ceutically acceptable Salts of the above-noted compounds. for combination with AHA's, either in the same formulation or a separate one, for improving skin resistance to damaging 0.043 C. Compositions of the Invention effects of Solar or ultraviolet radiation. 0044) The invention includes pharmaceutical composi 0050. One embodiment of the invention is a hydrogel tions for improving the net fidelity of DNA repair and for formulation, comprising an aqueous or aqueous-alcoholic US 2001/0044419 A1 Nov. 22, 2001 medium and a gelling agent, and a deoxyribonucleoside Zone (benzophenone-4; 2-hydroxy-4-methoxybenzophe Source. Suitable gelling agents include but are not limited to none-5-Sulfonic acid), octocrylene (2-ethylhexyl-2-cyano-3, methylcellulose, carboxymethylcellulose, hydroxypropylm 3-diphenylacrylate), octyl methoxycinnamate (2-ethylhexyl ethylcellulose, carbomer (carbopol), hypan, polyacrylate, p-methoxycinnamate), octyl Salicylate(2-ethylhexylsalicy and glycerol polyacrylate. late), homosalate (homomenthyl Salicylate), trolamine Sali cylate (triethanolamine Salicylate), phenylbenzimidazole 0051 Concentrations of gelling agents are selected Sulfonic acid, PABA (para-aminobenzoic acid), roXadimate according to their effect on Viscosity and pharmaceutical and (ethyl 4-bis hydroxypropyl aminobenzoate), lisadimate cosmetic propoerties. Suitable concentrations of a carbomer (glyceryl PABA), Padimate O (octyldimethyl PABA), men gelling agent, e.g. carbomer 934P range from 1 to 15%, thyl anthranilate, or Parsol 1789 (butyl methoxydibenzoyl advantageously 2 to 10% on a weight/weight basis. methane). 0.052 Liposomes are microscopic lipid vesicles which 0059 Alternatively, the compounds of the invention are can contain pharmacologically active agents either enclosed formulated in a base which is Suitable for application to the in the aqueous space within the vesicle or in the lipid skin prior to, or after, application of a SunScreen. This membrane itself, depending on the lipophilicity of the agent. embodiment of the invention permits the benefits of deox Liposomes are capable of delivering a pharmacologic agent yribonucleosides, in reducing consequences of UV damage through the Stratum comeum into deeper layers of the skin, and in attenuating photodynamic enhancement of cellular and are therefore Suitable vehicles for compounds and damage caused by Sunscreen agents themselves, to be compositions of the invention. obtained in conjunction with use of a wide variety of 0.053 Niosomes are lipid vesicles similar to liposomes commercial Sunscreen products. with membranes consisting largely of non-ionic lipids, Some forms of which are effective for transporting compounds 0060 D. Therapeutic Uses of the Compounds and Com acroSS the Stratum corneum. positions of the Invention 0054. In one embodiment of the invention, lipophilic acyl 0061 Reduction of Deleterious Consequences of Expo derivatives of deoxyribonucleosides, e.g. oleic or palmitic Sure of Skin to Ultraviolet or Solar Radiation acid esters of deoxyribonucleosides are incorporated into 0062) The compounds and composition of the invention, membranes of nioSome or lipoSome membranes, in addition when applied or administered before, during, or after expo to or instead of being enclosed within the vesicular mem Sure of the skin of a mammal to mutagenic radiation, have branes. the unexpected activity of improving the net fidelity DNA repair, thereby reducing mutation frequency, photoaging, 0055. Other agents which are advantageous for incorpo ration into a composition of the invention include corticOS and the chance of developing skin cancers. teroids, especially hydrocortisone in concentrations of 0.05 0063 DNA repair proceeds by several steps. A chemical to 1%, other anti-inflammatory corticosteroids at therapeu lesion in DNA, which can be caused by ultraviolet radiation, tically effective concentrations, topical anesthetics including ionizing radiation, free radicals, or chemical mutagens, is but not limited to benzocaine, lidocaine, and benzyl alcohol, detected by the DNA repair System. A Segment of nucle aloe Vera and aloe barbadensis, retinoids, antioxidants like otides including the damaged region is excised, generally Vitamins C and E, flavins, polyphenols (e.g. extracted from along with a number of Surrounding nucleotides. The green tea or black tea), allantoin, liposomal DNA repair excised Strand is then resynthesized from free deoxyribo enzymes, antibacterial agents (e.g. quaternary ammonium nucleotides, using the intact DNA Strand as a template. compounds, bacitracin, neomycin, polymyxin), Zinc salts, and methylxanthines. All of these listed agents have Some 0064. It is generally believed that improvement of repair utility in treating or attenuating various aspects of skin of DNA in skin cells (e.g. keratinocytes, melanocytes, injury or discomfort caused by ultraViolet radiation or fibroblasts) would require alteration in the activity of inflammatory skin conditions, and are therefore complemen enzymes or other proteins involved in the detection and excision of DNA lesions. Thus, there have been attempts to tary to the unique actions of the deoxyribonucleosides of the improve DNA repair in skin by delivering DNA repair invention. enzymes via topically-applied liposomes (U.S. Pat. No. 0056 Benzyl alcohol, which is known to have anesthetic (Yarosh et al., Cancer Res. 52:4227-31, 1992). and preservative properties, has the unexpected effect of improving aqueous Solubility of the relatively insoluble 0065 Far more important than the initial rate of excision purine deoxyribonucleosides, deoxyadenosine and deox of lesions is the fidelity or accuracy of repair. Agents which yguanosine; preferred concentrations of benzyl alcohol in accelerate the excision Step of DNA repair can actually topical formulations of deoxyribonucleosides are 0.5 to 5%. exacerbate damage if the cells are incapable of accurate This is very important in permitting high concentrations of repair Synthesis at a rate that matches the rate of excision of the deoxyribonucleosides of the invention to be stably damaged segments of DNA (Collins and Johnson, J. Cell incorporated into aqueous vehicles. Physiol. 99:125-137, 1979). 0066 Compounds and compositions of the invention, 0057 Sunscreens when applied to skin before, during or after exposure to Solar 0.058. The compounds of the invention are advanta or ultraViolet radiation or other environmental mutagens, geously incorporated into the same formulation as a UV reduces the mutation frequency otherwise induced by the absorbing chemical Sunscreen agent Such as: avobenzone mutagen (Example 1). This reduces the damage to cells (t-butyl dimethoxydibenzoylmethane), oxybenzone (ben caused by the mutagen, and reduces the chance of develop Zophenone-3), dioxybenzone (benzophenone-8), Sulisoben ment of Skin cancers, as shown in Examples 2 and 7. The US 2001/0044419 A1 Nov. 22, 2001 effect on genomic fidelity is in principle mediated by actual thus producing a short-lived excited Singlet State, return to improvement in the fidelity of repair in an individual cell or the ground State is accompanied by photon emission at by improvement of the elimination of cells with irreparable longer wavelengths that are Supposed to be leSS harmful than DNA damage. Either mechanism results in a net improve the incident radiation. Photon emission during the rapid ment in genomic fidelity in skin exposed to mutagens. return of a molecule from an excited Singlet State to a ground State is known as “fluorescence'. However, Sunscreens or 0067. The compounds of the invention have unantici other exogenous or endogenous UV-absorbing molecules pated benefits when used in combination with other agents can also be excited to longer-lived triplet States which can known to be useful in various aspects of skin care, including facilitate further reactions (the energy-emission that occurs but not limited to Sunscreens, methylxanthines, retinoids, during return of a molecule from an excited triplet State to DNA repair enzymes, exfoliants, and protease inhibitors, a ground State is known as “phosphorescence', and typically corticosteroids and nonsteroidal anti-inflammatory agents. occurs over a much longer time span than fluorescence). The 0068 The compounds and compositions of the invention, consequence is that Some UV-absorbing agents, especially when applied Soon enough, e.g. within about 3 days after those with a lowest triplet State that has a higher energy level exposure to ultraViolet or Solar radiation, improve the repair than the lowest triplet State of genomic DNA constituents, of cellular and macromolecular damage and improve net can absorb photons and actually exacerbate damage to DNA genomic fidelity, thereby reducing the chance of develop by direct or indirect energy transfer (e.g. from a triplet ment and Severity of macroscopically visible deleterious excited State) rather than by simple fluorescence, or photon consequences of Such exposure, including but not limited to emission at harmless wavelengths. photoaging, Sunburn Symptoms, actinic keratoses, Solar len tigines, “age Spots”, and skin cancer, e.g. basal cell carci 0074 Benzophenone, a close structural analog of oxy noma, Squamous cell carcinoma, melanoma. The com benzone, increases the yield of Strand breaks and pyrimidine pounds and compositions of the invention are also optionally dimers in UV-irradiated DNA, and is known to produce free applied before or during exposure to Solar or ultraViolet radicals upon irradiation with UV light (Charlier et al., radiation to shorten the time gap between damage and onset Photochemistry and Photobiology, 15:527-536, 1972). The of repair enhancement by the compounds of the invention. results presented in Examples 3, 4, 5 and 6 indicate that a Similar phenomenon also occurs with approved SunScreen 0069 Treatment of skin with compounds and composi ingredients, and that the deoxyribonucleosides of the inven tions of the invention results in a reduced chance of devel tion attenuate this deleterious consequence of Sunscreen use. opment of skin cancers and other deleterious consequences of exposure to Solar or UV radiation like photaging even 0075) When present in concentrations sufficient to block when the compounds of the invention are applied even after access of UV radiation to cells, Sunscreen agents are pro irradiation, e.g. after unintended exposure to potentially tective. However, if present in Very low concentrations, damaging doses of Solar radiation. This type of activity is not insufficient to adequately block UV transmission, photon shared by conventional SunScreens or agents which might absorbing agents, including common Sunscreen ingredients, act by enhancing melanogenesis, which are useful only if can operate as energy-transfer molecules, efficiently trap applied before irradiation. ping UV energy and transferring it to cell components, either directly (photosensitization) or by catalyzing formation of 0070 Compounds and compositions of the invention are reactive oxygen radicals (photodynamic sensitization). furthermore useful for treating existing inflammatory or Thus, low concentrations of oxybenzone, for example, hyperproliferative skin lesions, especially those caused by enhance DNA damage induced by ultraViolet radiation, exposure to Sunlight or ultraViolet radiation, including but whereas higher concentrations, Sufficient to block UV acceSS not limited to actinic keratoses, Solar lentigines, and to the cells or their immediate microenvironment altogether, Wrinkles. Example 9 illustrates this activity in a patient, in protect against DNA damage (see Example 3). which topical application of a composition of the invention resulted in complete regression of an an existing actinic 0076. The expected activity in vivo is that an oxyben keratosis. Zone-containing Sunscreen would protect cells from damage if present in a layer Sufficient to block access of light to the 0071 Improvement of Sunscreen Activity and Attenua target cells altogether. However, at lower concentrations, tion of Photodynamic Enhancement of UV Damage by insufficient to prevent penetration of UV radiation to target SunScreens cells, and especially if Some oxybenzone has been absorbed 0.072 Sunscreens are typically designed and tested on the into the critical cell layers, either via passage through the basis of prevention of Sunlight-induced erythema. While Stratum corneum or through hair follicles, there may be erythema and its attenuation by SunScreens is an important potentiation of damage to DNA in vivo. In mice treated Short-term effect, reduction of erythema and inflammation topically with commercial Sunscreen containing oxyben by SunScreens does not necessarily mean that they produce Zone, effects consistent with this hypothesis are in fact a proportionate protection of DNA (or prevention of skin observed after exposure to UV (see Example 6). Classes of cancers and Some features of photoaging Secondary to DNA Sunscreens other than benzophenone derivatives also exac damage). Sunscreens are certainly useful in preventing Some erbate UV-induced damage to DNA when present at low manifestations of photoaging and UV-related carcinogen concentrations during irradiation. esis, but do not provide complete protection, and in Some 0077. A strain of transgenic mice (v-HA-ras transgenic Situations may actually exacerbate photoinjury by acting as TG.AC mice) which is very susceptible to a variety of photodynamic Sensitizers (see Example 3). carcinogens, including UV radiation has been developed 0.073 Chemical sunscreens are intended to act by absorb recently (Leder et al., Proc. Nat. Acad. Sci. USA, 87.9178 ing photons at mutagenic (or erythmogenic) wavelengths, 9182, 1990). In response to a relatively small exposure to US 2001/0044419 A1 Nov. 22, 2001

UV radiation, these mice reliably develop cutaneous papil psoralens, which are present in Some perfume oils (berga lomas within a few weeks. When a circular patch of com mot), and which are in fact used to enhance Sunlight-induced mercial Sunscreen is applied to the back of Such a mouse, the tanning and UV phototherapy of psoriasis through exacer center of the protected region does in fact have lower bation of cellular injury. Many therapeutic drugs or their incidence of UV-induced papillomas than the unprotected metabolites are photochemically active chromophores Side, but often, along the margin of the applied Sunscreen, which can produce skin adverse reactions when a patient is there is a very high incidence (Sometimes higher than in exposed to Solar, visible, or ultraViolet radiation. Pigments unprotected areas) of papillomas (See Example 6). A layer and other light-absorbing constituents of cosmetics are also of Sunscreen Sufficient to block UV access to target cells is photochemically active chromophores which can exacerbate protective, but low concentrations, e.g. at the margin of a cellular photodamage. patch of Sunscreen) can act as photosensitizers increasing 0081. The deoxribonucleosides and related compounds the incidence of a UV-induced Skin cancer beyond that Seen of the invention (e.g. deoxyribonucleotides, oligonucle in completely “unprotected skin. Exposure of relevant skin otides, or DNA itself) are useful for attenuating cellular cells to low photosensitizing (rather than protective) con damage caused by excited light-absorbing molecules, centrations of Sunscreens clearly must occur during ordinary including exogenous photochemically active chromophores usage, e.g. at the margin of an applied patch, or as a like Sunscreens and cosmetic pigments, and also from protective layer is washed or worn off. endogenous chromophores like tryptophan, porphyrins, uro 0078. A benefit of deoxyribonucleosides or related com canic acid and melanin. pounds added to conventional Sunscreen formulations (or other cosmetics containing Sunscreens), beyond the Support 0082 Furthermore, since the photodynamic enhancement of DNA repair, is to Synergize with conventional SunScreen of DNA damage caused by benzophenone derivatives is in compounds by acting as energy Scavengers which trap part mediated by production of free radicals (Charlier et al., energy emitted by (or radicals produced by) Sunscreen Photochemistry and Photobiology, 15:527-536, 1972), com agents that is chemically similar to the cellular target, DNA. pounds and compositions of the invention are useful for Exogenous deoxyribonucleosides (in addition to their direct protecting the skin and mucosa from free radical damage, absorbance of UV energy) serve as “decoys” for energy whether or not the free radicals (e.g. hydroxyl radicals, captured by Sunscreen agents or other photoSensitizers that peroxide radicals, or lipoperoxide radicals) are initiated or would otherwise be transferred to cellular targets, including produced by photodynamic phenomena. Examples 3,4,5 and DNA. Thus, deoxyribonucleosides provide a dose-depen 6 provide evidence that the deoxyribonucleosides of the dent reduction in damage caused to cellular DNA by UV invention protect against DNA caused by free radicals. radiation in the presence of low concentrations of photo 0083. The deoxyribonucleosides and related compounds Sensitizing agents like oxybenzone or other Sunscreen agents of the invention are advantageously incorporated into the (Example 4). Same formulation as chemical SunScreen agents, which 0079 A defining characteristic of suitable energy scav include but are not limited to: avobenzone (t-butyl enging agents is that their lowest triplet energy State is equal dimethoxydibenzoylmethane), oxybenzone (benzophenone to or lower than that of DNA constituents in situ. Because of 3), dioxybenzone (benzophenone-8), Sulisobenzone (ben the Similarity of physicochemical properties of deoxyribo Zophenone-4), octocrylene (2-ethylhexyl-2-cyano-3,3- nucleosides (or deoxyribonucleotides, acyl deoxyribo diphenylacrylate), octyl methoxycinnamate (2-ethylhexyl nucleosides, or oligodeoxyribonucleotides) and genomic p-methoxycinnamate), octyl salicylate (2-ethylhexylsalicy DNA, the compounds of the invention are particularly late), homosalate (homomenthyl Salicylate), trolamine Sali Suitable as energy-Scavenging agents to protect genomic cylate (triethanolamine Salicylate), phenylbenzimidazole DNA from damage due to energy transfer from photosen Sulfonic acid, PABA (para-aminobenzoic acid), roXadimate Sitizers or photodynamic Sensitizers. In this embodiment, (ethyl 4-bis hydroxypropyl aminobenzoate), lisadimate ribonucleosides, ribonucleotides, oligoribonucleotides and (glyceryl PABA), Padimate O (octyldimethyl PABA), men acyl derivatives of ribonucleosides are within the Scope of thyl anthranilate, or Parsol 1789 (butyl methoxydibenzoyl the invention. Appropriate concentrations of Such Scaven methane). Such Sunscreen agents are present in formulations gers in a composition for topical application range from 0.1 at concentrations that are in accord with regulatory guide to 100 milligrams per milliliter (normalized to the amount of lines and Standard use. free nucleoside present in the case of nucleotides or oligo 0084. Similarly, in another embodiment of the invention, nucleotides or acyl derivatives of nucleosides). Advanta compounds of the invention are incorporated into cosmetics geously, Such Scavengers are present in concentrations rang containing photochemically active chromophores to mini ing from 0.1 to 20 mg/ml or especially 1 to 5 mg/ml. mize deleterious consequences of combined exposure of skin to Solar or ultraViolet radiation and Such cosmetic 0080. The problem of photodynamic enhancement of ingredients. damage to DNA extends beyond SunScreens. Other com pounds including endogenous molecules in the skin, can 0085 Alternatively, the compounds of the invention are absorb UV radiation at wavelengths that do not necessarily applied to the skin in a separate composition, e.g. a spray, directly damage DNA significantly, and transfer that energy lotion, roll-on, Stick, or gel, before or after a SunScreen to cellular targets including DNA, or generate free radicals product or cosmetic is applied. that damage cellular DNA. Examples of endogenous pho toSensitizing or photodynamically active skin constituents 0.086 Methylxanthines (photochemically active chromophores) include but are not 0087 Methylxanthines, such as caffeine, theophylline, limited to porphyrins, tryptophan, riboflavin, and melanin. aminophylline or isobutylmethylxanthine, have been pro Exogenous photodynamically active compounds include posed as "Sunless' tanning agents, which act by modulating US 2001/0044419 A1 Nov. 22, 2001 activity of the biochemical pathways involved in melano generally exposed to Sunlight, of patients receiving either genesis. Their proposed mechanism involves inhibition of oral or topical treatment with nonsteroidal anti-inflamma cyclic-AMP phosphodiesterase, thus enhancing the biologi tory agents, compositions of the invention overcome a cal activity of cyclic AMP in the pathways regulating deleterious consequence associated with this widely-used melanogenesis. Methylxanthines can enhance the produc class of drugs. tion of melanin in melanocytes, acting either alone or in combination with tanning Stimulants like ultraViolet or Solar 0.095 Ornithine Decarboxylase Inhibitors radiation. However, methylxanthines are also known to 0096. One consequence of UV irradiation of skin is an exacerbate DNA damage caused by ultraViolet radiation, increase in cell proliferation rate and in the activity of which has been attributed to an impairment of DNA repair enzymes necessary for cell proliferation, one of which is or disruption of cell cycle control mechanisms (Kastan et al., ornithine decarboxylase (ODC). Difluoromethylornithine Cancer Research, 51:6304-6311, 1991). (DFMO) is an inhibitor of ODC, which is necessary for 0088 Compounds of the invention are useful for reduc polyamine Synthesis, which in turn is necessary for DNA ing the deleterious effect of methylxanthines on DNA dam replication. DFMO is useful for reducing the incidence of age caused by exposure of skin to ultraViolet or Solar skin cancer and precancerous actinic lesions. Inhibition of radiation. The compounds of the invention thus improve the cell cycling after exposure to Solar or UV radiation may give Safety of skin tanning products that contain methylxanthines cells more time to repair DNA before mutagenic lesions are as active ingredients. Compounds of the invention are fixed by cell division, but cell cycle Stasis also generally applied either Separately or in the same formulation as the results in reduced levels of deoxyribonucleosides, which are methylxanthines. necessary for repair of DNA damage. Deoxyribonucleosides are therefore useful in conjunction with DFMO and other 0089 Exfoliants antiproliferative agents which act via mechanisms not directly involving induced depletion or imbalance of deox 0090 Cosmetics containing alpha-hydroxy acids (AHA) yribonucleotide pools. Deoxyribonucleosides of the inven Such as lactic acid, glycolic acid, citric acid, or malic acid are widely used. They have moisturizing and exfoliant proper tion are optionally incorporated into the same formulation as ties. Products containing high concentrations of AHA are DFMO (or other inhibitors of skin cell proliferation that act also used to produce more extreme “skin peels', in which through mechanisms other than impairment of DNA precur the Outer layers of the epidermis are removed, essentially by Sor Synthesis), or are applied in a separate formulation. means of a chemical burn. New epidermis growing in is 0097 Treatment of Skin During Exposure to Endogenous often Softer and Smoother than the skin layers that were Nitric Oxide removed. Beta-hydroxy acids like Salicylic acid are also 0.098 Nitric oxide (NO) is a biologically active mediator useful exfoliants. Retinoic acid is used for Similar purposes released by endothelial cells, macrophages and other cell through its exfoliant actions and through Stimulation of types, especially during inflammatory episodes. Nitric oxide epidermal cell turnover and alteration of epidermal metabo is also an important mediator of erythema associated with lism. UV exposure. Inhibitors of NO synthetase attenuate the 0.091 Exfoliants are reported to reduce the Sun-blocking increase in skin blood flow following exposure (Delicon capabilities of the Stratum corneum, and furthermore stantinos et al., J Cardiovasc Pharmacol 20 Suppl 12:S63-5, increase the permeability of the Skin to other agents. Thus, 1992; Deliconstantinos et al., BrJ Pharmacol 114(6):1257 there is a need for use of UV protection in conjunction with 65, 1995; Warren, FASEB Journal, 8(2):247-51, 1995). AHA's, yet the increased skin permeability produced by 0099 Nitric oxide is a potent inhibitor of the enzyme AHA requires caution in the Selection of Sun protection ribonucleotide reductase (RR), which is the key enzyme for agents, Since absorbed SunScreen agents can produce pho de novo synthesis of deoxyribonucleotides (Kwon et al., J todynamic enhancement of DNA damage. The compounds Exp Med 174(4):761-7, 1991; Lepoivre et al., J Biol Chem of the invention are effective in reducing deleterious con 269(34):21891-7, 1994) The antiproliferative effects of Sequences of exposure to Sunlight or ultraViolet radiation or nitric oxide are in part attributable to inhibition of ribonucle other environmental mutagens in Subjects using exfoliants, otide reductase. This can be beneficial in an inflammatory including but not limited to alpha-hydroxy acids, beta response to an infectious microorganism or a neoplasm, but hydroxy acids, and retinoids. is deleterious in cells in need of capability for DNA repair, 0092. Nonsteroidal Anti-inflammatory Agents e.g. skin exposed to inflammatory mediators elicited by UV 0093. Nonsteroidal anti-inflammatory agents are com exposure. The amounts of NO released from macrophages monly used for treatment of arthritis and other anti-inflam are sufficient to inhibit ribonucleotide reductase and thereby matory agents. Moreover, Some members of this class, e.g. induce cytostasis in neighboring cells. diclofenac (2,6-dichloro-phenyl-amino-phenylacetate) are 0100. The best-known inhibitor of RR, hydroxyurea under investigation as topical agents for reducing Some (HU), has structural Similarities to N-omega-hydroxy-1- aspects of skin photodamage. arginine, a physiological intermediate in NO production. 0094. One of the prototypical members of this class of Hydroxyurea can act as an NO-like nitroSating reactant drugs, acetaminophen, inhibits DNA repair after damage (LePoivre et al., J Biol Chem 2.69(34):21891-7, 1994). Both caused by UV radiation by inhibiting the enzyme ribonucle NO and hydroxyurea inhibit RR by quenching a tyrosyl otide reductase, which converts ribonucleoside diphosphates radical in the active Site of the enzyme. to deoxyribonucleoside diphosphates (Hongslo et al., 0101. A discovery first disclosed herein is that NO sen Mutagenesis, 8:423-429, 1993). By supplying deoxyribo sitizes cells to UV-induced DNA damage via mechanisms nucleosides to skin, especially to areas of the Skin that are that are reversible with exogenous deoxyribonucleosides US 2001/0044419 A1 Nov. 22, 2001

(see Example 8). Moreover, NO by itself, in the absence of 0106 Inhibition of DNA precursor synthesis by hydrox UV exposure, causes DNA damage that is prevented or yurea leads to enhanced activity and leakage of hydrolytic reversed by compounds of the invention (Example 8). lySOSomal enzymes which participate in extracellular dam age, e.g. in inflammatory skin conditions or photodamage 0102) The inflammatory response to UV exposure (which (Malec et al., Chem. Biol. Interact., 57:315-324, 1986). The also plays an important role in UV-associated immunoSup compounds of the invention prevent this component of pression), via inhibition of ribonucleotide reductase by NO, inflammatory tissue injury, especially when Such inhibition exacerbates the mutagenic and cytotoxic effects of UV of DNA precursor Synthesis is mediated by endogenously exposure. The Subject invention provides a means for com produced NO, which is functionally similar to hydroxyurea. pensating for this deleterious effect of NO. Exogenous deoxyribonucleosides, which enter into DNA metabolism 0107 A further contribution to genomic damage is that downstream of ribonucleotide reductase, compensate for exposure of cells to ultraViolet radiation results in the release reduced RR activity. NO, although deleterious in cells of enzymes, including deoxyribonuclease, from lysosomes. needing deoxyribonucleotides, has Some beneficial effects in Deoxyribonuclease II, which is present in lySOSomes, is an Sun-exposed skin. Inhibitors of NO production also impair endonuclease that can produce Strand breaks in nuclear DNA the healing of Skin damaged by exposure to excessive UV following lySOSome disruption. Leakage of enzymes from radiation, i.e., UV doses that produce a Sufficiently Severe lySOSomes also occurs in inflammatory conditions in gen Sunburn for wound healing processes to be called into action eral. The compounds and compositions of the invention are (Benrath et al., Neurosci Lett, 1995). Thus, NO is useful for useful for preserving genomic integrity after chromosomal tissue repair (probably by improving blood flow and perhaps damage caused by deoxyribonuclease released from lysos also by Stimulating growth factor release from macrophages omes in inflammatory skin conditions and after exposure of or keratinocytes), but may simultaneously exacerbate UV skin to Solar or ultraViolet radiation. related damage to DNA by reducing cellular capacities for 0108) By limiting some of the deleterious consequences production of deoxyribonucleotides for DNA repair. of skin inflammation, compounds and compositions of the 0103) NO-mediated inhibition of RR provides a mecha invention are useful as anti-inflammatory agents, and are nism for a disproportionate increase in DNA damage with optionally administered (either as separate compositions or, out a corresponding increase in other Symptoms of Sun advantageously, in the same formulation) in conjunction exposure during repeated exposure to Strong Sunlight. Topi with other topical or Systemic anti-inflammatory agents cal application of the compounds and compositions of the including but not limited to corticosteroids like hydrocorti invention provides a method for ameliorating this form of Sone and its congenerS. conditional hyperSensitivity. 0109 Similarly, compounds and compositions encom 0.104) NO participates in skin inflammatory reactions that passed by the invention are useful for treatment of mucosal do not necessarily involve exposure to Solar or ultraViolet inflammatory conditions, including but not limited to radiation. NO, which is released from activated macroph inflammatory bowel disease, ulcerative colitis, or Crohn's ages, is component of most inflammatory reactions. The disease, or mucositis anywhere in the gastrointestinal tract. compounds and methods of the invention provide a means of The preferred mode of treatment is by topical administra overcoming some deleterious effects of NO in inflammatory tion, in this case via enema or Suppository, for which skin conditions, including but not limited to pSoriasis, der purposes deoxyribonucleosides or other compounds of the matitis, allergic dermatitis, contact dermatitis (e.g. reactions invention are incorporated into Suitable vehicles. to poison ivy and poison oak), eczema and acne. In these 0110 Treatment of Skin and Mucosal Tissues Exposed to conditions, NO sensitizes some cell types to UV-induced Ionizing Radiation DNA damage by inhibiting deoxyribonucleotide synthesis. Compounds and compositions of the invention ameliorate 0111 Patients receiving therapeutic treatment (e.g. for this deleterious consequence of combined UV exposure and cancer) with ionizing (X-Ray or gamma) radiation can Suffer inflammatory skin conditions. Since NO and other endog damage to Skin overlying an internal tumor, leading to enous oxidants that cause DNA damage are present in deSquamation and poor healing. Compounds and composi inflammatory skin conditions like psoriasis or dermatitis tions of the invention are useful for treating damage to skin even in the absence of Significant exposure to UV radiation, and mucosal Surfaces caused by intentional or accidental the deoxyribonucleosides of the invention are useful for exposure to ionizing radiation. For treatment of Skin, com protecting genetic integrity of skin cells in inflammatory positions of the invention are applied topically before or after radiation treatment. For treatment of mucosal Surfaces, conditions. Compounds of the invention prevent or repair e.g. in the mouth, gastrointestinal tract, urethra or vagina, DNA damage caused by NO alone, without exposure to UV appropriate compositions of the invention are also applied radiation (Example 8). topically. Suitable compositions for treatment of mucosal 0105 Ribonucleosides are also useful for treating inflam Surfaces include gels, lotions, ointments, Suppositories, matory skin an mucosal conditions, in part by providing orally-administered capsules, pills or dragees, or Solutions. higher concentrations of the ribonucleotide Substrates for ribonucleoside reductase. Ribonucleosides (or ribonucleo 0112 E. Administration and Formulation of Compounds Side esters) in this context are used in the same ways that and Compositions of the Invention deoxyribonucleosides, advantageously in topically-applied 0113 Compounds of the invention are formulated in compositions, with ribonucleosides (or ribonucleoside pharmaceutically acceptable vehicles that deliver the com esters) incorporated in concentrations ranging from 0.1 to 20 pounds to the necessary cell populations in Skin at concen mg/ml, advantageously 1 to 5 mg/ml. Adenosine is particu trations adequate to accomplish the objectives of reducing larly useful for treatment of inflammatory conditions. mutation frequency and chance of developing cancer. US 2001/0044419 A1 Nov. 22, 2001

0114 Compositions of the invention are applied before, bowel disease, e.g. ulcerative colitis or Crohn's disease. For during, or after exposure to Sunlight or other mutagens. A treatment of mucositis in other parts of the gastrointestinal, lotion or hydrogel containing deoxyribonucleosides (0.1 to Such as the mouth, Standard pharmaceutically acceptable 10 mg/ml, advantageously 1 to 5 mg/ml) is applied to skin vehicles for that route of administration are used, e.g. as a thin film. The composition should be applied within mouthwashes or adherent hydrocolloids. about 48 or 72 hours after exposure to damaging doses of Sunlight or ultraViolet radiation, in order to provide Support 0120 F. Synthesis of the Compounds of the Invention for DNA repair prior to the first cell divisions after irradia 0121 Deoxyribonucleosides, being constituents of DNA, tion, although application before, during, or within 12 hours are present in all living organisms, and can therefore in after exposure to intense Sunlight is advantageous. Compo principle be extracted from a variety of Sources. In practice, Sitions of the invention are also effective when applied the most convenient biological Sources at present are fish before exposure to radiation or other mutagens, as long as milt, which contains a relatively high concentration of DNA. the deoxyribonucleosides So provided, or their anabolites, Fish milt sacs are homogenized, and the DNA therein is are available to cells in need of their beneficial effects at the partially purified and treated with deoxyribonucleases and time of exposure to a mutagen. Advantageously, composi phosphatases to degrade it to the level of nucleosides, which tions of the invention are applied within about 12 hours are then purified by chromatography and recrystallization. before exposure of the skin to a Solar radiation or other 0.122 Since mixtures of deoxyribonucleosides are used in mutagens. Some embodiments of the invention, purified deoxyribo 0115 Compositions of the invention are advantageously nucleosides are recombined in appropriate proportions. applied as a daily-use skin treatment, once to Several times Alternatively, deoxyribonuclosides are not separated from per day, especially on Sun-exposed parts of the body, or Sites each other during purification from other fish milt compo of inflammatory skin conditions. Exposure to Solar radiation nents (or other contaminants if the DNA is derived from leading to skin photodamage and photoaging is generally a other biological Sources); appropropriate quantities of indi cumulative process, involving repeated exposure to Sunlight, vidual deoxyribonucleosides are added to Such a mixture, if even daily, over a period of years. In this context, use of the necessary, to adjust the relative proportions of deoxyribo compounds and compositions of the invention to prevent or nucleosides. treat photodamage to the skin involves treatment of existing lesions due to prior Sun exposure, as well as prevention of, 0123 Deoxyribonucleosides can also be synthesized or attenuation of the Severity of, damage due to present and chemically from Simpler precursors. future exposure to Sunlight or other mutagens. 0124 Acyl derivatives of deoxyribonucleosides, as dis 0116. By improving repair of molecular damage to DNA closed in U.S. patent application Ser. No. 466,379, are useful as it occurs or before it is permanently established in the for 1) providing Sustained availability of deoxyribonucleo genome by cell division, or by preventing initial damage Sides due to gradual deacylation by nonspecific esterases in through energy Scavenging, compositions of the invention the skin, and 2) improved penetration through hydrophobic prevent or delay the manifestation of deleterious conse biological membranes or extracellular media, e.g. the inter quences of radiation, free radicals, or chemical mutagens, cellular lipids in the Stratum corneum of the epidermis. Such as grossly visible skin damage, photoaging, actinic 0.125. It will be obvious to the person skilled in the art keratoses, and Skin cancer. Thus, compositions and methods that other methods of Synthesis may be used to prepare the of the invention reduce the rate of appearance and the compounds of the invention. incidence of Signs of Skin photodamage, especially when 0.126 The following examples are illustrative, but not administered regularly, e.g. daily, or especially before, dur limiting of the methods and compositions of the present ing, or after exposure to Solar radiation. invention. Other Suitable modifications and adaptations of a 0117 Compositions of the invention are also useful for variety of conditions and parameters normally encountered promoting regression of established Sunlight-induced and in clinical therapy which are obvious to those skilled in the other inflammatory and hyperproliferative skin lesions, e.g. art are within the Spirit and Scope of this invention. actinic keratoses, contact dermatitis, psoriasis, eczema, or acne, or skin cancers like melanoma, basal cell carcinoma, EXAMPLE 1. or Squamous cell carcinoma. In Such conditions, topical gels, 0127 Post-irradiation topical deoxyribonucleosides creams, ointments or lotions are applied to the affected areas improve DNA repair in mouse skin after UVB exposure The once or twice per day as needed. incidence of mutations in Skin in response to ultraViolet 0118. In one embodiment of the invention, a composition radiation was determined using the “Big Blue' transgenic containing deoxyribonucleosides is applied to Skin prior to mouse test System. These mice carry approximately 40 application of a SunScreen, as an alternative to use of copies per cell of a lambda phage shuttle vector containing formulations containing both conventional SunScreens and a lad gene as a target for mutagenesis, as well as the lacI deoxyribonucleosides or related compounds of the inven promoter, the lac operator, and the Clac7, reporter gene. tion. 0128. Following exposure of the mice to ultraviolet radia 0119 For treatment of colon mucosal inflammation, e.g. tion and treatment with compounds of the invention or inflammatory bowel disease, compositions of the invention vehicle, genomic DNA from skin Samples is extracted and are administered as a Suppository or enema, approximately the shuttle vectors are packaged in lambda Virus heads. The once per day according to clinical need. Volumes of 10 to phage lambda Viruses containing the shuttle vectors are 500 ml of a Suitable enema composition containing deox plated on E. Coli with the color reagent X-Gal, which turns yribonucleosides are Suitable for treatment of inflammatory blue when enzymatically altered by galactosidase, the prod US 2001/0044419 A1 Nov. 22, 2001 uct of the Clac7 gene. Viruses containing nonmutated lacI (0.136) In 9 mice exposed to UV-B radiation (0.3-1.25 genes produce white plaques, mutation of the lacI gene kJ/m X3 q2d) and treated after irradiation with vehicle results in blue plaques. The mutation frequency is deter (propylene glycol), a total of 35 papillomas were found 4 mined by counting the relative numbers of white and blue weeks after exposure. plaques. 0.137 Among 7 mice exposed to the same doses of UV-B 0129. Stratagene BigBlueTM transgenic mice (n=7/group) radiation and treated with deoxyribonucleosides (4 mg/ml of were shaved and then irradiated the next day with UVB each major deoxyribonucleoside in propylene glycol) after radiation (85% of energy at 313 nm), 1.25 kJ/m2. irradiation, only 1 papilloma was observed (Table 2). 0130 Deoxyribonucleosides (“dNsides”; 4 mg/ml each of deoxyadenosine, deoxycytidine, deoxyguanosine, and TABLE 2 thymidine) or vehicle (propylene glycol) were applied topi Topical deoxyribonucleosides reduce UV-induced tumorigenesis cally 30 minutes after irradiation and again each day for 5 days. Control: 3.89 tumors/mouse dNside-treated: 0.14 tumors/mouse 0131 Mice were sacrificed on day 5 after irradiation. DNA was extracted from dorsal (irradiated) and ventral (non-irradiated) skin, packaged into lambda phage and 0.138. The beneficial effect of deoxyribonucleosides in plated on E. Coli along with X-Gal. Colonies with mutations reducing UV-induced tumorigenesis must be due to were blue. >200,000 colonies were counted in each group. improvement of repair phenomena or cellular proofreading, or to inhibition of tumor promotion, and not to prevention of initial damage, Since the deoxyribonucleosides were applied after irradiation. This observation indicates that deoxyribo TABLE 1. nucleoside treatment after (or presumably also during) expo Topical deoxyribonucleosides reduce mutation sure to LWV-B radiation has important inhibitory effects on frequency in UV-irradiated skin tumorigenesis, as a consequence of improved maintenance of genomic fidelity. Spontaneous mutation frequency (nonirradiated skin): Average: 4.5 x 10 0.139. This antitumorigenic effect of topical deoxyribo Mutation frequency in UVB-irradiated skin: nucleosides even when applied after irradiation is particu larly Surprising in View of the reported efficacy of deoxyri Total Increment due to UV bonucleosides in accelerating wound healing (U.S. Pat. No. Control 34.9 x 10 30.4 x 10 5,246,708), since other growth factors known to accelerate dNsides 7.8 x 10 3.3 x 10 wound healing, like platelet-derived growth factor (PDGF) or transforming growth factor beta (TGF-B), act as tumor promoters. 0.132. As shown in Table 1, post-irradiation treatment with topical deoxyribonucleosides reduced the incidence of 0140. The antitumorigenic effect of deoxyribonucleo mutations caused by UV-B radiation by a factor of nearly 10 Sides in this experiment is also unexpected in View of the beneficial effect of deoxyribonucleosides on Survival of cells in this experiment (30.4x10 versus 3.3x10) in culture when the deoxyribonucleosides are applied after EXAMPLE 2 exposure of the cells to ultraViolet or ionizing radiation. Prevention of apoptosis of damaged cells would be expected 0.133 Post-irradiation Treatment with Topical Deoxyri to increase the likelihood of tumor development, as occurs bonucleosides Prevents Development of UV-induced Papil in animals with defective p53-related mechanisms. lomas in v-Ha-ras Transgenic TG.AC Mice 0134 Example 1 demonstrated that post-irradiation topi EXAMPLE 3 cal treatment can reduce the frequency of UV-induced 0141 Low Concentrations of Oxybenzone Exacerbate mutations in a reporter gene in “Big Blue' transgenic mice, UTV-induced Damage to DNA through Support and improvement of DNA repair processes. One of the consequences of reduced mutation frequency in 0.142 Confluent human fibroblasts in T25 flasks were response to a carcinogen like UV radiation should be a washed 3 times with HBSS (Hank's Balanced Salt Solution) and incubated with vehicle or with various concentrations of reduction of UV-induced tumorigenesis. oxybenzone (OB) for 2 hours. Media was aspirated and cells 0135 A strain of transgenic mice has been developed were covered with a 1 mm layer of HBSS and irradiated which is extremely Sensitive to carcinogens. It permits rapid from above with UV-B (50J/m). The medium was aspirated determination of carcinogenic potential of various chemical and cells were incubated for three hours with 2 mM hydrox agents and other treatments. Normal mice require repeated yurea. Medium was again aspirated, and cells were exposure to ultraViolet radiation over a number of weeks in trypsinized with 0.25% trypsin/EDTA. Cells were centri order to reliably develop skin tumors. In contrast, V-Ha-ras fuged at 4 C., resuspended in 50 microliters of HBSS and TG.AC transgenic mice can develop tumors rapidly after a incubated at room temperature with 200 microliters of 1N Single exposure, or Small number of exposures to UV NaOH for 15 minutes. DNA damage (single strand breaks) radiation. was assessed by alkaline Sucrose gradient centrifugation. US 2001/0044419 A1 Nov. 22, 2001

"Nucleoid position' in the Sucrose gradient is proportional EXAMPLE 5 to the number of DNA single strand breaks. 0147 Effect of Individual Versus Combined Deoxribo 0143 UV irradiation without oxybenzone results in a nucleosides on Photodynamically-enhanced, UV-induced three to four-fold increase in the nucleoid position over DNA Damage baseline (Table 3). Oxybenzone at the higher concentrations 0.148 Human fibroblasts were prepared and treated as in tested, 20 and 200 micromolar, protected cellular DNA Example 4, and the effects of individual deoxyribonucleo against damage from the UV irradiation, as the nucleoid sides on photodynamically enhanced DNA damage (2 uM position for these groups is close to that of non-irradiated oxybenzone) were determined. Individual deoxyribonucleo cells. However, cells exposed to 2 micromolar OB display Sides were tested at concentrations of 20 uM, and the substantially more damage than cells irradiated with no OB combination of all four deoxyribonucleosides contained at all; nucleoid position values are 10-fold greater than those thymidine, deoxycytidine, deoxyadenosine and deoxygua of non-irradiated cells. Thus, oxybenzone Strongly enhances nosine at 20 uM each. DNA damage when present at low concentrations, whereas it protects cells at higher concentrations. In practice, even TABLE 5 under conditions of proper SunScreen use, there will always Effect of individual versus combined be areas of skin exposed to low, potentially deleterious deoxyribonucleosides on UV-induced DNA damage concentrations of Sunscreen, either at the edge of a patch of DNA Strand Breaks applied Sunscreen, or as an applied layer wears off over the Group (Breaks/47 MDa) course of a day. Control (no UV) O UV + vehicle 325 TABLE 3 UV + thymidine 13 UV + deoxycytidine .12 LOW concentrations of Oxybenzone enhance UV-induced DNA damage UV + deoxyadenosine 17 UV + deoxyguanosine 17 Group Nucleoid Position (mm) UV + dNsides (20 uM each) O1 No UV 3 UV 50 J/m2 11 UV 50 J/m2 + 2 uMOB 32 0149. As shown in Table 5, each of the individual deox UV 50J/m2 + 20 uMOB 4. yribonucleosides attenuated DNA damage; the combination UV 50 J/m2 + 200 uMOB 3 of all four deoxyribonucleosides was substantially more effective than any individual compound.

EXAMPLE 4 EXAMPLE 6 0144) Deoxribonucleosides Attenuate Photodynamic 0150 Sunscreen-induced Exacerbation of UV-induced Enhancement of DNA Damage Caused by Oxybenzone Tumorigenesis and its Prevention with Deoxyribonucleo Sides 0145 Confluent human fibroblasts were exposed to 2 micromolar oxybenzone (OB), as in Example 3, prior to 0151. A circular patch of commercial sunscreen (Cop exposure to UV-B radiation (50 J/m). Different flasks of pertone SPF 8, which includes oxybenzone) was applied to cells also were exposed to increasing concentrations of the backs of TG.AC mice prior to exposure to 125 J/mi deoxyribonucleosides. Cells were processed for determina UV-B radiation. tion of nucleoid position in a Sucrose density gradient, a 0152 Around the circular margin of the area that was measure of DNA single strand breaks. covered with Sunscreen during irradiation, 8 papillomas per 0146). As shown in Table 4, deoxyribonucleosides pro mouse were present at 10 dayS. 3 tumors per mouse were duce a dose-dependent reduction in the yield of DNA Single observed in animals not treated with Sunscreen. In mice strand breaks induced by UV exposure plus 2 uM OB. exposed to UV radiation after application of the same Deoxyribonucleosides at 2 uM slightly reduce DNA dam commercial Sunscreen to which deoxyribonucleosides had age; at 200 micromolar, the deoxyribonucleosides almost been added, no papillomas were elicited (Table 6). completely abrogate the DNA damage. TABLE 6 TABLE 4 Deoxyribonucleosides attenuate tumorigenesis Deoxyribonucleosides attenuate photodynamic enhancement of in in mice treated with UV plus sunscreen DNA damage caused by UV plus oxybenzone UV Radiation 3 tumors/mouse Sunscreen + UV radiation: 8 tumors/mouse Group Nucleoid Position (mm) Sunscreen containing dNsides + UV radiation: Otumors/mouse UV + 2 uM OB 56 UV + 2 uM OB + 2 uM dNsides 47 UV + 2 uM OB + 2.0 uM dNsides 23 UV + 2 uM OB + 100 uM dNsides 22 0153. Thus, at the margin of the applied patch of Sun UV + 2 uM OB + 200 uM dNsides 7 Screen where the SunScreen concentration diminishes in a rapid gradient toward Zero, tumorigenic UV damage in fact appears to be enhanced rather than reduced by the SunScreen US 2001/0044419 A1 Nov. 22, 2001 agents, leading to formation of more premalignant papillo Groups of cells were also exposed to deoxyribonucleosides mas than on Skin not treated with Sunscreen at all. of the invention before, or before and after, exposure to NO 0154 Addition of deoxyribonucleosides to a commercial or NO+UV radiation; control groups were treated identically Sunscreen abrogated the deleterious effect of the SunScreen except for addition of deoxyribonucleosides. on tumorigenesis. 0169 DNA was extracted from cells and subjected to pulsed field gel electrophoresis to determine the incidence of EXAMPLE 7 DNA strand breaks. 0155 Effect of Individual Deoxyribonucleosides Versus 0170 As shown in Table 8, NO alone produced a sig a Combination on UV-induced Tumorigenesis nificant incidence of DNA strand breaks, which were further 0156 Thirty v-Ha-ras TGAC mice aged twelve weeks increased by exposure to UV radiation. The deoxyribo were, shaved and Subjected to ultraviolet radiation, 1.25 nucleosides of the invention Strongly reduced the incidence kJ/mon days 0, 6, 8 and 11, at a dose rate of 12.5 W/m2. of DNA strand breaks caused by NO, as well as those caused O157 Mice were divided into groups of five animals each by the combination of NO plus UV radiation. and treated, beginning 30 minutes after irradiation, with: TABLE 8 0158 1. Vehicle (propylene glycol) 0159 2. dNsides-Deoxyribonucleosides (4 mg/ml Deoxyribonucleosides attenuate Nitric Oxide-induced DNA damage each of deoxyadenosine, deoxycytidine, deoxygua DNA Strand Breaks nosine, and thymidine) Groups (breaks/105 MDa) Control O.OO 0160 3. dC-Deoxycytidine (4 mg/ml) NO O.13 NO + UV 50J/m2 O.21 0161 4. dG-Deoxyguanosine (4 mg/ml) NO + UV 300 J/m2 O.51 0162 5. dA-Deoxyadenosine (4 mg/ml) NO+ dNsides before and after UV O.OO NO + UV 50 J/m' + dNsides before/after UV O.O3 0163 6.dT Thymidine (4 mg/ml) NO + UV 300 J/m2 + dNsides before/after UV O.O3 NO+ dNsides after UV O.O6 0164. Animals were observed for 8 weeks, during which NO + UV 50 J/m' + dNsides after UV O.O6 time the development of papillomas was observed. NO + UV 300 J/m2 + dNsides after UV O.O3

TABLE 7 EXAMPLE 9 Effect of individual versus combined deoxyribonucleosides on UV-induced tumorigenesis 0171 Treatment of Existing Actinic Keratosis with Topi Papillomas/mouse at the end of 8 weeks: cal Deoxyribonucleosides Vehicle 18 dNsides O.O 0172 Actinic keratosis (AK) is a form of intradermal dC O.25 neoplasia, a Sunlight-induced lesion which can progreSS to dG 1.O dA O.67 become Squamous cell carcinoma. A 46 year-old woman dT 1.6 with very light skin presented with a persistent rose colored, rough-Surfaced lesion 5x10 mm in size on her forehead. The lesion was diagnosed as an actinic keratosis by a board 0165. As shown in Table 7, the mixture of all four certified dermatologist, who prescribed topical 5-fluorou deoxyribonucleosides provided the best activity in terms of racil for its treatment. Since 5-fluororacil, a Standard treat prevention of tumor development. Deoxycytidine also pro ment for AK, can produce pain and unsightly skin erosion Vided protection; deoxyadenosine and deoxyguanosine were which takes Several weeks to heal and is also potentially less protective but nonetheless had activity. Thymidine did genotoxic, the Subject elected to first try to treat the AK with not have significant protective actions. topical deoxyribonucleosides (4 mg/ml each of deoxycyti dine, deoxyadenosine, deoxyguanosine, and thymidine) in a EXAMPLE 8 hydrogel formulation, of which 0.1 ml was applied to the 0166 Nitric Oxide Causes and Enhances DNA Damage lesion twice daily. Within 10 days, the AK on her forehead by a Deoxyribonucleoside-reversible Mechanism had begun to visibly regreSS, and after three weeks, there was no trace of the AK, nor was there any Scar or discomfort 0167 Nitric oxide is a mediator of UV-induced erythema, during or after treatment. 5-fluorouracil treatment and other and is also present in inflammatory skin conditions. NO is painful or invasive measures like excision or freezing with mutagenic, and may exacerbate UV-induced DNA damage, liquid nitrogen were thus not needed. The AK has not in part by inhibiting ribonucleotide reductase, the enzyme recurred after more than Six months. responsible for conversion of ribonucleotides to deoxyribo nucleotides. EXAMPLE 10 0168 Human melanocytes were exposed to 160 uM 0173 Formulation of a Composition of the Invention DETA NONOate (Alexis Corporation, Cat # 430-014 M005), a reagent which spontaneously produces nitric oxide 0.174. A suitable clinical formulation of a composition of when exposed to water. Cells were exposed to NO alone, or the invention comprises the following ingredients. The batch NO plus increasing doses of UV radiation (50 and 300 J/m). Size is optionally Scaled to any desired Volume. US 2001/0044419 A1 Nov. 22, 2001 14

8. A method as in claim 3 wherein said at least one deoxyribonucleoside is administered in a vehicle containing 1. 2'-Deoxycytidine Hydrochloride 0.4640 g from 0.1 to 10 milligrams per milliliter of each deoxyribo 2. 2'-Deoxyadenosine Monohydrate 0.4290 g nucleoside. 3. 2'-Deoxyguanosine Monohydrate 0.4270 g 9. A method as in claim 3 wherein said at least one 4. Thymidine 0.4000 g deoxyribonucleoside is administered in a vehicle containing 5. Edetate Disodium Dihydrate, USP 0.0100 g from 1.0 to 5 milligrams per milliliter of each deoxyribo 6. Benzethonium Chloride, USP 0.0250 g 7. Butylated Hydroxytoluene, NF 0.0005 g nucleoside. 8. Carbomer 934P, NF 0.7200 g 10. A method as in claim 3 wherein said at least one 9. Glycerin, USP 10.000 g deoxyribonucleoside is a mixture of deoxycytidine, deoxy 10. Alcohol, USP 5.000 g adenosine, deoxyguanosine, and thymidine. 11. Diethylene glycol monoethyl ether 10.000 g 12. NaOH, NF (1 N Solution) as needed for pH adjustment 11. A method as in claim 10 wherein said deoxyribo 13. HCl, NF (1 N Solution) as needed for pH adjustment nucleoside is administered in a vehicle containing from 0.1 14. Purified Water, USP q.S. to 100.00 g to 10 milligrams per milliliter of each deoxyribonucleoside. 12. A method as in claim 10 wherein said deoxyribo nucleoside is administered in a vehicle containing from 1 to 0175 Preparation: 5 milligrams per milliliter of each deoxyribonucleoside. 13. A method for reducing mutation frequency in skin of 0176 1. Item 8 (Carbomer 934P) is dissolved in 35 a mammal exposed to a mutagen comprising administering g of Purified Water. to Said Skin a Source of at least one deoxyribonucleoside. 0177 2. Item 5 is dissolved into the solution. 14. A method as in claim 13 wherein Said mutagen is Selected from the group consisting of ultraViolet or Solar 0.178 3. Into a separate container, Items 9, 10, 11 are radiation, a chemical mutagen, a free radical, or ionizing mixed with 15 g Purified Water. radiation. 0179 4. Items 1, 2, 3, 4, 6, and 7 are dissolved in the 15. A method as in claim 13 wherein said source is Solution of Step 3, Selected from the group consisting of at least one free or acyl deoxyribonucleoside. 0180 5. The solutions of step 2 and 4 are mixed 16. A method as in claim 13 wherein said source is together. Selected from the group consisting of a deoxyribonucleotide, 0181 6.8.85g of Item 12 are added to the solution an oligodeoxyribonucleotide, and a poly deoxyribonucle of step 5. otide. 17. A method as in claim 13 wherein said Source of at least 0182 7. The resulting gel is blended for 15 minutes, one deoxyribonucleoside is administered in a vehicle at a and the pH is adjusted to 7.0+/-0.5 with Item 12 or concentration of from 1.0 to 20 milligrams per milliliter. 13 as needed. 18. A method as in claim 15 wherein said at least one deoxyribonucleoside is free or acyl deoxycytidine. 0183 8. The final weight of the batch is brought to 19. A method as in claim 15 wherein said at least one 100 g with Purified Water followed by gentle blend deoxyribonucleoside is free or acyl deoxyadenosine. ing for 15 minutes. 20. A method as in claim 15 wherein said at least one 0184 The foregoing is intended as illustrative of the deoxyribonucleoside is free or acyl deoxyguanosine. present invention but not limiting. Numerous variations and 21. A method as in claim 15 wherein Said at least one modifications may be effected without departing from the deoxyribonucleoside is free or acyl thymidine. true Spirit and Scope of the invention. 22. A method as in claim 15 wherein Said at least one deoxyribonucleoside is administered in a vehicle containing from 0.1 to 10 milligrams per milliliter of each deoxyribo What is claimed is: nucleoside. 1. A method of improving DNA repair in the skin or 23. A method as in claim 15 wherein said at least one mucosa of a mammal comprising administering to Said deoxyribonucleoside is administered in a vehicle containing mammal at least one deoxyribonucleoside, deoxyribonucle from 1.0 to 5 milligrams per milliliter of each deoxyribo otide, or oligodeoxyribonucleotide. nucleoside. 2. A method as in claim 1 wherein Said at least one 24. A method as in claim 15 wherein Said at least one deoxyribonucleoside is administered topically. deoxyribonucleoside is a mixture of free or acyl deoxycy 3. A method as in claim 1 wherein Said at least one tidine, deoxyadenosine, deoxyguanosine, and thymidine. deoxyribonucleoside is Selected from the group consisting of 25. A method as in claim 24 wherein said at least one deoxycytidine, deoxyadenosine, deoxyguanosine, and thy deoxyribonucleoside is administered in a vehicle containing midine. from 0.1 to 10 milligrams per milliliter of each deoxyribo 4. A method as in claim 3 wherein Said at least one nucleoside. deoxyribonucleoside is deoxycytidine. 26. A method as in claim 24 wherein Said at least one 5. A method as in claim 3 wherein said at least one deoxyribonucleoside is administered in a vehicle containing deoxyribonucleoside is deoxyadenosine. from 1 to 5 milligrams per milliliter of each deoxyribo 6. A method as in claim 3 wherein Said at least one nucleoside. deoxyribonucleoside is deoxyguanosine. 27. A method for reducing the chance of developing skin 7. A method as in claim 3 wherein said at least one cancer in a mammal due to exposure to a mutagen compris deoxyribonucleoside is thymidine. ing administering to the skin of Said mammal a Source of at US 2001/0044419 A1 Nov. 22, 2001

least one deoxyribonucleoside wherein Said Source is admin 48. A method as in claim 43 wherein said at least one istered Such that Said at least one deoxyribonucleoside is deoxyribonucleoside is administered in a vehicle containing present on Skin of Said mammal during or after exposure to from 0.1 to 10 milligrams per milliliter of each deoxyribo Said mutagen in an amount Sufficient to reduce the delete nucleoside. rious consequences of Said exposure. 49. A method as in claim 43 wherein said at least one 28. A method as in claim 27 wherein said source is deoxyribonucleoside is administered in a vehicle containing administered within 3 days after exposure to a mutagen. from 1.0 to 5 milligrams per milliliter of each deoxyribo 29. A method as in claim 28 wherein said mutagen is nucleoside. Selected from the group consisting of Solar, ultraViolet, or 50. A method as in claim 43 wherein said at least one ionizing radiation. deoxyribonucleoside is a mixture of deoxycytidine, deoxy 30. A method as in claim 27 wherein said source is adenosine, deoxyguanosine, and thymidine. Selected from the group consisting of at least one free or acyl 51. A method as in claim 50 wherein said at least one deoxyribonucleoside. deoxyribonucleoside is administered in a vehicle containing 31. A method as in claim 27 wherein said source is from 0.1 to 10 milligrams per milliliter of each deoxyribo Selected from the group consisting of a deoxyribonucleotide, nucleoside. an oligodeoxyribonucleotide, and a poly deoxyribonucle 52. A method as in claim 50 wherein said at least one otide. deoxyribonucleoside is administered in a vehicle containing 32. A method as in claim 27 wherein said Source of at least from 1 to 5 milligrams per milliliter of each deoxyribo one deoxyribonucleoside is administered in a vehicle at a nucleoside. concentration of from 1.0 to 20 milligrams per milliliter. 53. A method for reducing the chance of developing 33. A method as in claim 30 wherein said at least one actinic keratoses in a mammal comprising administering to deoxyribonucleoside is free or acyl deoxycytidine. the skin of Said mammal a Source of at least one deoxyri bonucleoside. 34. A method as in claim 30 wherein said at least one deoxyribonucleoside is free or acyl deoxyadenosine. 54. A method as in claim 53 wherein said source is Selected from the group consisting of a deoxyribonucleo 35. A method as in claim 30 wherein said at least one Side, a deoxyribonucleotide, an oligodeoxyribonucleotide, deoxyribonucleoside is free or acyl deoxyguanosine. and an acyl derivatives of deoxyribonucleoside. 36. A method as in claim 30 wherein said at least one 55. A method of reducing the deleterious consequences of deoxyribonucleoside is free or acyl thymidine. photosensitization or photodynamic Sensitization on the skin 37. A method as in claim 30 wherein Said at least one of a mammal caused by an endogenous or exogenous deoxyribonucleoside is administered in a vehicle containing photochemically active chromophore comprising adminis from 0.1 to 10 milligrams per milliliter of each deoxyribo tering to Said skin an energy Scavenging agent with a lowest nucleoside. triplet State energy less than or equal to that of nucleobases 38. A method as in claim 30 wherein said at least one in DNA. deoxyribonucleoside is administered in a vehicle containing 56. A method as in claim 55 wherein said exogenous from 1.0 to 5 milligrams per milliliter of each deoxyribo chromophore is a SunScreen agent. nucleoside. 57. A method as in claim 56 wherein said Sunscreen agent 39. A method as in claim 30 wherein said at least one is selected from the group consisting of avobenzone (t-butyl deoxyribonucleoside is a mixture of free or acyl deoxycy dimethoxydibenzoylmethane), oxybenzone (benzophenone tidine, deoxyadenosine, deoxyguanosine, and thymidine. 3), dioxybenzone (benzophenone-8), Sulisobenzone (ben 40. A method as in claim 39 wherein said at least one Zophenone-4; 2-hydroxy-4-methoxybenzophenone-5-Sul deoxyribonucleoside is administered in a vehicle containing fonic acid), octocrylene (2-ethylhexyl-2-cyano-3,3- from 0.1 to 10 milligrams per milliliter of each deoxyribo diphenylacrylate), octyl methoxycinnamate (2-ethylhexyl nucleoside. p-methoxycinnamate), octyl salicylate (2-ethylhexylsalicy 41. A method as in claim 39 wherein said at least one late), homosalate (homomenthyl Salicylate), trolamine Sali deoxyribonucleoside is administered in a vehicle containing cylate (triethanolamine Salicylate), phenylbenzimidazole from 1 to 5 milligrams per milliliter of each deoxyribo Sulfonic acid, PABA (para-aminobenzoic acid), roXadimate nucleoside. (ethyl 4-bis hydroxypropyl aminobenzoate), lisadimate 42. A method as in claim 27 wherein Said skin cancer is (glyceryl PABA), Padimate O (octyldimethyl PABA), men Selected from the group comprising basal cell carcinoma, thyl anthranilate, or Parsol 1789 (butyl methoxydibenzoyl melanoma, and Squamous cell carcinoma. methane) 58. A method as in claim 55 wherein said energy scav 43. A method for reducing the rate of skin photoaging in enging agent is Selected from the group consisting of DNA, a mammal comprising administering to the skin of Said an oligodeoxyribonucleotide, a ribonucleoside, a deoxyri mammal at least one deoxyribonucleoside. bonucleoside, a ribonucleotide, a deoxyribonucleotide, an 44. A method as in claim 43 wherein said at least one acyl deoxyribonucleoside, and an acyl ribonucleoside. deoxyribonucleoside is deoxycytidine. 59. A method as in claim 58 wherein said energy scav 45. A method as in claim 43 wherein said at least one enging agent is administered in a vehicle containing 0.1 to deoxyribonucleoside is deoxyadenosine. 20 mg/ml. 46. A method as in claim 43 wherein said at least one 60. A method as in claim 58 wherein said energy scav deoxyribonucleoside is deoxyguanosine. enging agent is a mixture comprising free or acylated 47. A method as in claim 43 wherein said at least one deoxycytidine, deoxyadenosine, deoxyguanosine, and thy deoxyribonucleoside is thymidine. midine. US 2001/0044419 A1 Nov. 22, 2001

61. A method as in claim 60 wherein said mixture is 84. A method as in claim 78 wherein said at least one administered in a vehicle containing from 0.1 to 10 milli deoxyribonucleoside is administered in a vehicle containing grams per milliliter of each deoxyribonucleoside. from 0.1 to 10 milligrams per milliliter of each deoxyribo 62. A method for reducing the deleterious consequences nucleoside. of exposure of a mutagen to the Skin of a mammal com 85. A method as in claim 78 wherein said at least one prising administering to Said mammal at least one deoxyri deoxyribonucleoside is administered in a vehicle containing bonucleoside, deoxyribonucleotide, or oligodeoxyribo from 1.0 to 5 milligrams per milliliter of each deoxyribo nucleotide. nucleoside. 63. A method as in claim 62 wherein Said mutagen is Solar 86. A method as in claim 78 wherein said at least one or ultraViolet radiation. deoxyribonucleoside is a mixture of free or acyl deoxycy 64. A method as in claim 62 wherein Said mutagen is tidine, deoxyadenosine, deoxyguanosine, and thymidine. ionizing radiation. 87. A method as in claim 86 wherein said at least one 65. A method as in claim 62 wherein Said mutagen is a deoxyribonucleoside is administered in a vehicle containing free radical. from 0.1 to 10 milligrams per milliliter of each deoxyribo 66. A method as in claim 62 wherein said at least one nucleoside. deoxyribonucleoside is administered topically. 88. A method as in claim 86 wherein said at least one 67. A method as in claim 62 wherein said at least one deoxyribonucleoside is administered in a vehicle containing deoxyribonucleoside is Selected from the group consisting of from 1 to 5 milligrams per milliliter of each deoxyribo deoxycytidine, deoxyadenosine, deoxyguanosine, and thy nucleoside. midine. 68. A method as in claim 67 wherein said at least one 89. A method of treating skin inflammation in a mammal deoxyribonucleoside is deoxycytidine. comprising administering to inflamed skin a Source of at 69. A method as in claim 67 wherein said at least one least one deoxyribonucleoside or ribonucleoside. deoxyribonucleoside is deoxyadenosine. 90. A method as in claim 89 wherein said skin inflam 70. A method as in claim 67 wherein said at least one mation is Selected from the group consisting of dermatitis, deoxyribonucleoside is deoxyguanosine. pSoriasis, eczema, and acne. 71. A method as in claim 67 wherein said at least one 91. A method as in claim 89 wherein said skin inflam deoxyribonucleoside is thymidine. mation is due to exposure to Solar or ultraViolet radiation. 72. A method as in claim 67 wherein said at least one 92. A method as in claim 89 wherein said Source of at least deoxyribonucleoside is administered in a vehicle containing one deoxyribonucleoside or ribonucleoside is Selected from from 0.1 to 10 milligrams per milliliter of each deoxyribo the group consisting of DNA, an oligodeoxyribonucleotide, nucleoside. a ribonucleoside, a deoxyribonucleoside, a ribonucleotide, a 73. A method as in claim 67 wherein said at least one deoxyribonucleotide, an acyl deoxyribonucleoside, and an deoxyribonucleoside is administered in a vehicle containing acyl ribonucleoside. from 1.0 to 5 milligrams per milliliter of each deoxyribo 93. A method as in claim 92 wherein said Source of at least nucleoside. one deoxyribonucleoside or ribonucleoside is administered 74. A method as in claim 67 wherein said at least one in a vehicle containing from 0.1 to 20 mg/ml. deoxyribonucleoside is a mixture of deoxycytidine, deoxy 94. A method as in claim 92 wherein said Source of at least adenosine, deoxyguanosine, and thymidine. one deoxyribonucleoside is a mixture comprising free or 75. A method as in claim 74 wherein said deoxyribo acylated deoxycytidine, deoxyadenosine, deoxyguanosine, nucleoside is administered in a vehicle containing from 0.1 and thymidine. to 10 milligrams per milliliter of each deoxyribonucleoside. 95. A method as in claim 89 wherein said Source of at least 76. A method as in claim 74 wherein said deoxyribo one deoxyribonucleoside is deoxycytidine. nucleoside is administered in a vehicle containing from 1 to 96. A method as in claim 89 wherein said Source of at least 5 milligrams per milliliter of each deoxyribonucleoside. one ribonucleoside is adenosine. 77. A method of reducing the deleterious consequences of 97. A method as in claim 89 wherein said adenosine is exposure of skin to endogenously produced nitric oxide administered in a vehicle containing from 0.1 to 10 mg/ml. comprising administering to Said Skin a Source of at least one 98. A method of treating mucosal inflammation in a deoxyribonucleoside. mammal comprising administering to Said mucosal inflam 78. A method as in claim 77 wherein said Source of at least mation a Source of at least one deoxyribonucleoside or one deoxyribonucleoside is Selected from the group consist ribonucleoside. ing of a deoxyribonucleoside, a deoxyribonucleotide, an 99. A method as in claim 98 wherein said mucosal skin oligodeoxyribonucleotide, or an acyl deoxyribonucleoside. inflammation is Selected from the group consisting of 79. A method as in claim 77 wherein said Source of at least inflammatory bowel disease, ulcerative colitis, Crohn's dis one deoxyribonucleoside is administered in a vehicle at a ease, Stomatitis or mucositis. concentration of from 1.0 to 20 milligrams per milliliter. 100. A method as in claim 98 wherein said source of at 80. A method as in claim 78 wherein said at least one least one deoxyribonucleoside or ribonucleoside is Selected deoxyribonucleoside is free or acyl deoxycytidine. from the group consisting of DNA, an oligodeoxyribonucle 81. A method as in claim 78 wherein said at least one otides, a ribonucleoside, a deoxyribonucleoside, a ribonucle deoxyribonucleoside is free or acyl deoxyadenosine. otide, a deoxyribonucleotide, an acyl deoxyribonucleoside, 82. A method as in claim 78 wherein said at least one and an acyl ribonucleoside. deoxyribonucleoside is free or acyl deoxyguanosine. 101. A method as in claim 100 wherein said Source of at 83. A method as in claim 78 wherein said at least one least one deoxyribonucleoside is administered in a vehicle deoxyribonucleoside is free or acyl thymidine. containing from 0.1 to 20 mg/ml. US 2001/0044419 A1 Nov. 22, 2001

102. A method as in claim 100 wherein said Source of at 117. A composition as in claim 109 wherein said at least least one deoxyribonucleoside is a mixture comprising free one deoxyribonucleoside is a mixture comprising free or acylated deoxycytidine, deoxyadenosine, deoxygua deoxycytidine, deoxyadenosine, deoxyguanosine, and thy nosine, and thymidine. midine. 103. A method as in claim 98 wherein said Source of at 118. A composition as in claim 117 wherein each deox least one deoxyribonucleoside is deoxycytidine. yribonucleoside is present in a concentration of 0.1 to 10 104. A method of reducing the chance of developing skin milligrams per milliliter. cancer in a mammal due to exposure to Solar or ultraViolet 119. A composition as in claim 117 wherein each deox radiation comprising topically administering a composition yribonucleoside is present in a concentration of 1 to 5 comprising a Sunscreen agent and an energy Scavenging milligrams per milliliter. agent. 120. A composition as in claim 109 wherein said agent 105. A method as in claim 104 wherein said Sunscreen that enhances penetration is Selected from the group con agent is Selected from the group consisting of avobenzone sisting of ethanol, isopropanol, azone (1-dodecylazacyclo (t-butyl dimethoxydibenzoylmethane), oxybenzone (ben heptan-2-one), oleic acid, linoleic acid, propylene glycol, Zophenone-3), dioxybenzone (benzophenone-8), Sulisoben hypertonic glycerol, lactic acid, glycolic acid, citric acid, Zone (benzophenone4; 2-hydroxy-4-methoxybenzophenone malic acid, and diethylene glycol monoethyl ether. 5-Sulfonic acid), octocrylene (2-ethylhexyl-2-cyano-3,3- 121. A composition as in claim 120 wherein Said agent diphenylacrylate), octyl methoxycinnamate (2-ethylhexyl that enhances penetration is diethylene glycol monoethyl p-methoxycinnamate), octyl salicylate (2-ethylhexylsalicy ether. 122. A composition as in claim 121 wherein Said dieth late), homosalate (homomenthyl Salicylate), trolamine Sali ylene glycol monoethyl ether is present in a concentration of cylate (triethanolamine Salicylate), phenylbenzimidazole 2 to 20%. Sulfonic acid, PABA (para-aminobenzoic acid), roXadimate 123. A composition as in claim 121 wherein said dieth (ethyl 4-bis hydroxypropyl aminobenzoate), lisadimate ylene glycol monoethyl ether is present in a concentration of (glyceryl PABA), Padimate O (octyldimethyl PABA), men 5 to 15%. thyl anthranilate, or Parsol 1789 (butyl methoxydibenzoyl 124. A composition as in claim 109 wherein said com methane) position is a hydrogel. 106. A method as in claim 104 wherein said energy 125. A composition as in claim 124 wherein the gelling Scavenging agent is Selected from the group consisting of agent for Said hydrogel is Selected from the group consisting DNA, an oligodeoxyribonucleotide, a ribonucleoside, a of methylcellulose, carboxymethylcellulose, hydroxypropy deoxyribonucleoside, a ribonucleotide, a deoxyribonucle lmethylcellulose, carbomer, Hypan, polyacrylate, and glyc otide, an acyl deoxyribonucleoside, and an acyl ribonucleo erine polyacrylate. Side. 126. A composition as in claim 124 wherein the gelling 107. A method as in claim 106 wherein said energy agent for Said hydrogel is carbomer. Scavenging agent is administered in a vehicle containing 127. A composition as in claim 126 wherein the concen from 0.1 to 20 mg/ml. tration of Said carbomer is 2 to 10%. 108. A method as in claim 106 wherein said energy 128. A composition comprising Scavenging agent is a mixture comprising free or acylated deoxycytidine, deoxyadenosine, deoxyguanosine, and thy a) deoxyguanosine and deoxyadenosine and midine. b) benzyl alcohol. 109. A composition comprising 129. A composition as in claim 128 wherein the concen tration of said benzyl alcohol is between 0.1 and 5%. a) at least one deoxyribonucleoside and 130. A composition as in claim 128 wherein the concen b) an agent that enhances penetration of Said at least one trations of deoxyguanosine and deoxyadenosine are between deoxyribonucleoside into the Skin. 0.1 and 10 mg/ml. 110. A composition as in claim 109 wherein said skin is 131. A composition as in claim 128 further containing human skin. deoxycytidine and thymidine. 111. A composition as in claim 109 wherein said at least 132. A composition as in claim 131 wherein the concen one deoxyribonucleoside is deoxycytidine. trations of deoxycytidine and thymidine are between 0.1 and 10 mg/ml. 112. A composition as in claim 109 wherein said source 133. A composition comprising a methylxanthine and of at least one deoxyribonucleoside is deoxyadenosine. Source of at least one deoxyribonucleoside. 113. A composition as in claim 109 wherein said source 134. A composition as in claim 133 wherein said source of at least one deoxyribonucleoside is deoxyguanosine. of at least one deoxyribonucleoside is Selected from the 114. A composition as in claim 109 wherein said source group consisting of at least one deoxyribonucleoside and at of at least one deoxyribonucleoside is thymidine. least one acyl derivative of a deoxyribonucleoside. 115. A composition as in claim 109 wherein said at least 135. A composition comprising one deoxyribonucleoside is present in a concentration of from 0.1 to 10 milligrams per milliliter of each deoxyribo a) a Sunscreen agent nucleoside. b) an energy Scavenging agent. 116. A composition as in claim 109 wherein said at least 136. A composition as in claim 135 wherein said Sun one deoxyribonucleoside is present in a concentration of Screen agent is Selected from the group consisting of from 1.0 to 5 milligrams per milliliter of each deoxyribo avobenzone (t-butyl dimethoxydibenzoylmethane), oxyben nucleoside. Zone (benzophenone-3), dioxybenzone (benzophenone-8), US 2001/0044419 A1 Nov. 22, 2001

Sulisobenzone (benzophenone-4; 2-hydroxy-4-methoxyben 147. A method as in claim 145 wherein said Source is Zophenone-5-Sulfonic acid), octocrylene (2-ethylhexyl-2- Selected from the group consisting of a deoxyribonucleotide, cyano-3,3-diphenylacrylate), octyl methoxycinnamate an oligodeoxyribonucleotide, and a poly deoxyribonucle (2-ethylhexyl p-methoxycinnamate), octyl salicylate (2-eth otide. ylhexylsalicylate), homosalate (homomenthyl salicylate), trolamine Salicylate (triethanolamine Salicylate), phenylben 148. A method of reducing the rate of development of skin Zimidazole Sulfonic acid, PABA (para-aminobenzoic acid), photodamage in a mammal due to exposure to Solar or roxadimate (ethyl 4-bis hydroxypropyl aminobenzoate), ultraViolet radiation comprising topically administering a lisadimate (glyceryl PABA), Padimate O (octyldimethyl composition comprising a SunScreen agent and an energy PABA), menthyl anthranilate, and Parsol 1789 (butyl meth Scavenging agent. oxydibenzoylmethane). 149. A method as in claim 148 wherein said energy 137. A composition as in claim 135 wherein said energy Scavenging agent is Selected from the group consisting of Scavenging agent is Selected from the group consisting of a DNA, an oligodeoxyribonucleotide, a ribonucleoside, a Source of at least one deoxyribonucleoside. deoxyribonucleoside, a ribonucleotide, a deoxyribonucle 138. A composition as in claim 137 wherein said source otide, an acyl deoxyribonucleoside, and an acyl ribonucleo of at least one deoxyribonucleoside is Selected from the Side. group consisting of a deoxyribonucleoside, a deoxyribo 150. A method for inducing regression of inflammatory or nucleotide, an oligodeoxyribonucleotide, or an acyl deox hyperproliferative skin lesions due to exposure to Solar or yribonucleoside. ultraViolet radiation comprising topically administering a 139. A composition as in claim 138 wherein said at least one deoxyribonucleoside is a mixture comprising free or composition comprising a Source of at least one deoxyribo acylated deoxycytidine, deoxyadenosine, deoxyguanosine, nucleoside or deoxyribose. and thymidine. 151. A method as in claim 150 wherein said skin lesion is 140. A composition as in claim 138 wherein each deox Selected from the group comprising actinic keratosis, Solar yribonucleoside is present in a concentration of from 0.1 to lentigines, pSoriasis, dermatitis, eczemea, melanoma, basal 10 milligrams per milliliter. cell carcinoma, and Squamous cell carcinoma. 141. A composition as in claim 138 wherein said at least 152. A method as in claim 150 wherein said Source of at one deoxyribonucleoside is free or acyl deoxycytidine. least one deoxyribonucleoside is Selected from the group 142. A composition as in claim 141 wherein Said free or consisting of a deoxyribonucleoside, a deoxyribonucleotide, acyl deoxycytidine is present in a concentration of from 0.1 an oligodeoxyribonucleotide, or an acyl deoxyribonucleo to 100 milligrams per milliliter. Side. 143. A composition as in claim 135 wherein Said energy 153. A method as in claim 152 wherein said at least one Scavenging agent is Selected from the group consisting of a deoxyribonucleoside is a mixture comprising free or acy Source of at least one ribonucleoside. lated deoxycytidine, deoxyadenosine, deoxyguanosine, and 144. A composition as in claim 143 wherein Said Source thymidine. of at least one deoxyribonucleoside is Selected from the 154. A method as in claim 152 wherein each deoxyribo group consisting of a ribonucleoside, a ribonucleotide, an nucleoside is present in a concentration of from 0.1 to 10 oligoribonucleotide, or an acyl ribonucleoside. milligrams per milliliter. 145. A method for reducing the rate of development of 155. A method as in claim 152 wherein said at least one skin photodamage in a mammal exposed to Solar or ultra deoxyribonucleoside is free or acyl deoxycytidine. Violet radiation comprising administering to the skin of Said mammal a Source of at least one deoxyribonucleoside 156. A method as in claim 155 wherein said free or acyl wherein Said Source is administered Such that Said deoxyri deoxycytidine is present in a concentration of from 0.1 to bonucleoside is present on skin of Said mammal during or 100 milligrams per milliliter. after exposure to Said radiation in an amount Sufficient to 157. A method as in claim 150 wherein said deoxyribose reduce the deleterious consequences of Said exposure. is present in a topical formulation in a concentration of 0.1 146. A method as in claim 145 wherein said Source is to 100 millimolar. Selected from the group consisting of at least one free or acyl deoxyribonucleoside.