Compounds and composition for DNA protection

Abstract

The present disclosure relates to compounds and composition for DNA protection. More particularly, the invention discloses compound and composition to reduce DNA damage, to repair the damaged DNA and to enhance the DNA repair wherein such DNA damage relate to ageing related pathologies or diseases.

Description

D E S C R I P T I O NCOMPOUNDS AND COMPOSITION FOR DNA PROTECTIONTECHNICAL FIELD

[0001] The present disclosure relates to compounds and composition for DNA protection. More particularly, the invention discloses compound and composition to reduce DNA damage, to repair the damaged DNA and to enhance the DNA repair wherein such DNA damage relate to ageing related pathologies or diseases. BACKGROUND

[0002] Aging is a complex and multifaceted biological process that involves the gradual deterioration of a variety of mechanisms and cellular and molecular components over time[1]. The accumulation of these defects along with the lack of countermeasures, lead to the quick decline of cellular functions. Several interconnected mechanisms contribute to aging process, influencing cellular function and tissue integrity[1]. The accumulation of those contributions results in phenotypical onset, that have been originated due to months of molecular and cellular deterioration. This phenotypical onset result on age-related features such as cognitive changes in memory, executive function, processing speed; physical changes in musculoskeletal, sensory and cardiovascular changes; among others[2]. Those manifestations are the result of cellular senescence, mitochondrial dysfunction, telomere shortening, epigenetic changes, protein homeostasis and also, but not only, DNA damage and mutation induction[3][4][5].

[0003] The accumulation of damage in the DNA results in the formation of mutations that can interfere or originate other problems such does mentioned early. Organisms developed different ways in order to overcome this problem and different molecular pathway are responsible for correct or adjust different types of damage on the DNA[6]. Humans have several mechanisms that are responsible to deal with damage within transcriptional coupled damage, global genome damage, for the excision of nucleotides, of base pairs, double strand and single strand breaks, being originated from different sources[6]. For that, different proteins are constitutively being express in order to keep DNA homeostasis,protein such as CSB, CSA, XPC, XPA, XPB, TFIIH, among other proteins that are responsible to guide different damage repair pathways[7][8].

[0004] CSB and CSA proteins are strongly associated with the transcriptional coupled – nucleotide excision repair (TC-NER) pathway[7]. These protein act in one of the first steps of the mechanism and the absence of these proteins result in the accumulation of damage on genes. Mutations in these proteins result in non-functional proteins, which compromises the pathway and lead to the development of a disease called Cockayne syndrome[7][9]. Furthermore, this is not exclusive of the TC-NER pathway, the lack of the protein XPC, directly relate with the global genome – nucleotide excision repair (GG-NER) pathway results in a similar phenotypical disease called xeroderma pigmentosum

[0010] . None of those proteins are exclusive of the pathways previously mentioned, CSB for instance also act on base excision repair (BER), double strand break repair (DSR) and others[8]. However, the lack of repair mechanisms in both cases resulted in age related disease in which its carriers have a short life expectancy presenting age-related phenotypical features since early age[7][9]

[0010] .

[0005] These facts are disclosed in order to illustrate the technical problem addressed by the present disclosure. GENERAL DESCRIPTION

[0006] The present invention relates to novel compounds and compositions designed for mitigating DNA damage, facilitating DNA repair processes, and enhancing overall DNA repair. More particularly, the present disclosure discloses compound and composition to reduce DNA damage, to repair the damaged DNA and to enhance the DNA repair wherein such DNA damage relate to ageing related pathologies or diseases namely addressing genetic mutations related to the CSB (Cockayne syndrome group B) / CSA (Cockayne syndrome group A) gene and associated ageing-related pathologies or diseases.

[0007] The present disclosure aims to address the underlying mechanisms of DNA damage, repair, and maintenance to combat the effects of CSB / CSA mutations, ultimately reducing the impact of age-related diseases associated with DNA damage.

[0008] The disclosed compounds and compositions exhibit potent DNA protective properties, promoting DNA repair mechanisms, and augmenting the overall efficiency ofrepairing DNA lesions induced by various environmental factors, genetic predispositions, and ageing. The present disclosure demonstrate efficacy in minimizing the detrimental effects of CSB / CSA mutations on DNA integrity, thereby presenting promising prospects in managing and potentially ameliorating diseases or conditions linked to CSB / CSA mutations and ageing-related pathologies. Geroprotective effects

[0009] A compound with geroprotective effects, as depicted and described herein.

[0010] The use of the compound for protecting biological systems from environmental stressors.

[0011] In the present disclosure was used the specific feature of the presence or absence of damage on the DNA to perform a pharmacological screening, test and identify geroprotective, namely through the correction or reduction of DNA damage

[0011] . The present disclosure also shows the expression of the protein CSB in order to understand which drugs promote DNA damage repair through CSB dependent and independent pathways. It was performed a single cell gel electrophoresis in order to assess DNA damage in a population of cells, and western blot analysis for CSB, to detect alterations in the level of this protein.

[0012] 1. An aspect of the present disclosure relates to compound sfor use in the prevention or treatment of DNA repair-deficiency disorder caused by ageing related diseases, disorders or symptoms, wherein the compound is selected from a list consisting of: Hydralazine hydrochloride, Mafenide hydrochloride, Iproniazide phosphate, (S,+) Camptothecine, Tolfenamic acid, Fenspiride hydrochloride, Mebendazole, Tropicamide, Riluzole hydrochloride, Clemizole hydrochloride, Atracurium besylate, Sulfacetamide sodic hydrate, Heptaminol hydrochloride, Sulfathiazole, Hydrochlorothiazide, Idoxuridine, Isoflupredone acetate, Amiloride hydrochloride dihydrate, Amprolium hydrochloride, Betazole hydrochloride, Isoxicam, Ticlopidine hydrochloride, Dicyclomine hydrochloride, Levodopa, and combinations thereof.

[0013] In an embodiment, said use is for the prevention or treatment of DNA repair- deficiency disorder, disease, or symptoms caused by ageing related diseases, disorders or symptoms from the group comprising: Cockayne Syndrome type A, Cockayne Syndrome type B, Xeroderma pigmentosum, Hutchinson-Gilford progeria syndrome.

[0014] In an embodiment, the compound of the present disclosure may be use for reducing DNA damage associated with ageing-related pathologies and / or CSB mutations.

[0015] In an embodiment, the compound of the present disclosure may be use for enhancing DNA repair mechanisms influenced by mutations in the CSB gene, CSA gene, XPC gene, XPA gene, XPB gene, TFIIH gene; preferably CSB gene.

[0016] These genes are crucial in maintaining DNA integrity and repairing damage caused by various environmental factors, particularly UV radiation from sunlight. Namely: CSB gene - Cockayne syndrome group B protein gene, associated with Cockayne syndrome, a rare genetic disorder characterized by sensitivity to sunlight and premature aging. CSA gene - Cockayne syndrome group A protein gene, also associated with Cockayne syndrome. XPC gene: -Xeroderma pigmentosum complementation group C protein gene, involved in nucleotide excision repair, which helps repair DNA damaged by UV radiation. XPA gene - Xeroderma pigmentosum complementation group A protein gene, also involved in nucleotide excision repair. XPB gene- Xeroderma pigmentosum complementation group B protein gene, a subunit of the TFIIH complex involved in DNA unwinding during nucleotide excision repair. TFIIH gene - Transcription factor IIH gene, a complex of proteins involved in both transcription and DNA repair, including roles in nucleotide excision repair.

[0017] In an embodiment, said mutation is in a gene selected from de group consisting of: CSB, CSA, XPC, XPA, XPB, TFIIH; preferably CSB gene.

[0018] In an embodiment, the compound of the present disclosure may be used in the prevention or disorders / diseases related with mutation of the CSB gene or CSA gene; preferably CSB gene.

[0019] In an embodiment, the compound of the present disclosure may be used in the prevention or treatment of Premature Aging symptoms, UV Sensitivity Disorders, Neurological Disorders, Cockayne syndrome.

[0020] Another aspect of the present disclosure relates to a pharmaceutical composition for use in the prevention or treatment of DNA repair-deficiency disorder caused by ageing related diseases, disorders, or symptoms, comprising at least one compound disclosed in the present disclosure in a therapeutical amount and a pharmaceutically acceptable carrier, adjuvant, excipient, emulsification agent or mixtures thereof.

[0021] In an embodiment, the compound, at least one of the compounds of the present disclosure may be use in the treatment of ageing related diseases, comprising for use in the treatment of ageing related diseases, comprising at least one compound according to any in a therapeutical amount and a pharmaceutically acceptable carrier, adjuvant, excipient, emulsification agent or mixtures thereof.

[0022] The present disclosure relates to use a compound for the manufacture of a medicament for the treatment of prevention or treatment of DNA repair-deficiency disorder caused by ageing related diseases, disorders or symptoms; wherein at least a compound is the compound disclosed in the present disclosure.

[0023] A method for treating or preventing DNA repair-deficiency disorder caused by ageing related diseases, disorders or symptoms in a subject, wherein the method comprising administering at least one of the compounds of any of the present disclosure to the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The following figures provide preferred embodiments for illustrating the disclosure and should not be seen as limiting the scope of invention.

[0025] Figure 1: Graphic representation with standard deviation of the values of CSB expression in each condition. In ANOVA analysis none of the drugs presented significant difference on the CSB expression when compared with the control group (P>0.05).

[0026] Figure 2: Immunoblot analysis of CSB expression in HEK-293T cells. Total protein was extracted from cells and western blot was performed with 50 microg of protein per sample in 7.5% acrylamide gel. Blot was stained with polyclonal antibody (A301-345A, BETHYL) for CSB / ERCC6 (168 kDa) and polyclonal antibody for B-tubulin (#0000184173, merch) (55 kDa). No statistical different was observed in any condition compared with the control group (P>0.05).

[0027] Figure 3. Among the all the drugs that have been tested on this study, 31,5% of them presented significant results regarding reduce DNA damage (namely the 23 drugs of the present disclosure) and 68,5% did not present any significant variation. DETAILED DESCRIPTION

[0028] Approximately 23 mL of Agarose Normal Melting Point (ANMP) in PBS 1x was prepared along with 6 mL of agarose Low Melting (ALMP) in PBS 1x and let at 60°C in a water bath. A piece of Gel Bound Film was cut with 7,9 x 11,9 cm. 1,5 mL of ANMP was added to a multiwell lid and the Gel Bound Film with the hydrophobic side facing down was place on top of it and let it stay for 5 minutes at room temperature. On top of the film 13,5 ml of ANMP were added and immediately the microporous gel template was placed on top of it and let it cooldown for 15 minutes without move or touch it. Approximately 8x10^4 cells were seeded in T-flasks 75cm^2 and kept at 37ºC and 5% CO2, until reach 70% confluence. A cell suspension with approximately 4x10^6 cells was prepared in 3,85 ml of DMEM. Cells were seeded in a 96 well microporous gel at a concentration of 40.000 cells per well. Cells were incubated at 37°C for 1 hour in order to precipitate in the microwells. After the incubation, the gel was washed with PBS 1x and then covered with ALMP warmed to 37°C and incubated at room temperature for 10 minutes, followed by 5 minutes at 4°C. Cells were treated with drugs diluted DMSO (1 microM) followed by a final dilution in DMEM to achieve a concentration of 0,2 microM. Cells were exposed for 30 minutes at 4°C to the different compounds and after rinsed with PBS 1x. After, the gel was submerged on lysis solution (2.5 M NaCl, 100 mM Na2EDTA, 10 mM Tris-Base) (pH = 10) overnight at 4°C. On the following day, gel was rinsed with PBS 1x and place in the comet assay tina apparatus with the gel side facing down. Alkaline Electrophoresis buffer (0.3 M NaOH, 1 mM Na2EDTA) (pH>13) was added until it covers the gel. The gel was then equilibrated in Alkaline Electrophoresis buffer for 1 hour at 4°C and an electrophoresis was performed at constant 21 V for 50 minutes. The volume buffer was adjusted in order to obtain a 0,3 A. After the electrophoresis, the gel was immerged on Tris-Base 0,4M for 15 minutes twice at 4°C followed by incubation for 30 minutes at Tris-Base 20 microM at 4°C. After the gel was DNA stained with SYBR gold 1x for 1 hour in the dark at 4°C.Microscopy image acquisition

[0029] Microscopy images were acquired with an Imager Z2 (Zeiss) microscope with motorized stage using 5x objectives with tiles. Images were acquired using the Axiocam 705 mono camera (Zeiss) resorting to Zen 3.4 software (Zeiss). Images were then uploaded on ImageJ software (version 2.15.0) for western blot bands densitometric analyze and Comet Assay analysis with OpenComet plugin (v1.3) Western Blot

[0030] Samples were treated and diluted in RIPA buffer [20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1% NP-40, 1% sodium deoxycholate] and Protease Inhibitor Cocktail (made in-house). 50 microg protein / lane was separated on 7,5% acrylamide gels. The electrophoresis was performed at 80V for 20 minutes followed by 2:25 hours at 120V in running buffer (0.02M Tris + 0.02M Bicine + 3.46mM SDS, pH 8,3). A PVDF membrane was activated in a methanol bath for 20 seconds, washed in distilled water for 5 minutes, followed by immersion in CAPS 1x methanol 10x for 15 minutes. The acrylamide gel was submerged in CAPS 1X methanol 10x for 15 minutes during the same time in order to equilibrate the gel. Both gel and membrane were mounted in the transfer device followed by wet transfer at 4°C at 750mA for 2 hours and 15 minutes. After transfer membrane was blocked with 5% milk in TBS-T with agitation for 1 hour at room temperature. Primary antibody against CSB rabbit was diluted 1:2000 on blocking solution (5% milk in TBS-T) and primary antibody against B-tubulin was diluted in 1:5000 on blocking solution (5% milk in TBS-T), both were incubated overnight at 4°C with agitation. Membrane was washed three times in TBS-T 1x, incubated with conjugated antibody diluted in blocking solution (5% milk in TBS-T) 1:10000 at room temperature with agitation for 2 hours. By chemiluminescence with the assistance of Enhanced Chemiluminscence (ECL prime) results were obtained in Chemidoc (Molecular Imager, Chemidoc XRS+, Bio rad). Compounds

[0031] Different compounds were obtained from Prestwick Chemical Library® of 1520 FDA- EMA approved compounds, which represent a diverse array of pharmacological activities. 49 Drugs (active ingredient) such as Hydralazine hydrochloride

[0012] , Mafenidehydrochloride

[0013] , Iproniazide phosphate

[0014] , (S,+) Camptothecine

[0015] , Tolfenamic acid

[0016] , Fenspiride hydrochloride

[0017] , Mebendazole

[0018] , Tropicamide

[0019] , Riluzole hydrochloride

[0020] , Clemizole hydrochloride

[0021] , Atracurium besylate

[0022] , Sulfacetamide sodic hydrate

[0023] , Heptaminol hydrochloride

[0024] , Sulfathiazole

[0025] , Hydrochlorothiazide

[0026] , Idoxuridine

[0027] , Isoflupredone acetate

[0028] , Amiloride hydrochloride dihydrate

[0029] , Amprolium hydrochloride

[0030] , Betazole hydrochloride

[0031] Isoxicam

[0032] , Ticlopidine hydrochloride

[0033] , Dicyclomine hydrochloride

[0034] , Levodopa

[0035] Meticrane

[0036] , Hydroflumethiazide

[0037] , Azaguanine-8

[0038] , Allantoin

[0039] , Acetazolamide

[0040] , Diflunisal

[0041] , Procaine hydrochloride

[0042] , Moxisylyte hydrochloride

[0043] , Amyleine hydrochloride

[0044] , Lidocaine hydrochloride

[0045] , Ethosuximide

[0046] , Mafenide hydrochloride

[0047] , Nitrofurantoin

[0048] , Tranylcypromine hydrochloride

[0049] , Aceclofenac

[0049] , Sulfamethoxazole

[0050] , Mephenesin

[0051] , Phenformin hydrochloride

[0052] , Pargyline hydrochloride

[0053] , Pentolinium bitartrate

[0054] , Aminopurine

[0055] , 6-benzyl

[0056] , Lomefloxacin hydrochloride

[0057] , Orphenadrine hydrochloride

[0058] , Dilazep dihydrochloride

[0059] , Tolbutamide

[0060] Nimesulide, Salbutamol

[0061] , Prilocaine hydrochloride

[0062] , (S,+) Camptothecine

[0053] , Ranitidine hydrochloride

[0054] , Tiratricol

[0055] , Flufenamic acid

[0056] , Flumequine

[0057] , Tibolone

[0058] , Trimethoprim

[0059] , Piroxicam

[0060] , Gemfibrozil

[0061] , Mefexamide hydrochloride

[0062] , Fenbufen

[0063] , Ketoprofen

[0064] , Norfloxacin

[0065] , Antimycin A

[0066] , Nifenazone

[0067] , Griseofulvin

[0068] , Phentolamine hydrochloride

[0069] , Etodolac

[0070] , Scopolamin-N-oxide hydrobromide

[0071] , Ofloxacin

[0072] were tested) and only 23 compound demonstrated activity in the prevention or treatment of DNA repair-deficiency disorder caused by ageing related diseases.

[0032] . Among those drugs twelve class of drugs were tested, varying from allergology treatments to oncologic drugs treatment (table 1

[0033] Table 1. Classification of tested drugs within name, therapeutical class, and literature reference.Drug Therapeutic class Reference

[0034] In the present disclosure, statistical analyses were performed using both GraphPad Prism (Graph-Pad Software, La Jolla, USA) and SPSS (IBM SPSS Statistic software v29). ANOVA test, multiple comparison tests and optimal binding tests were performed with a minimal significant threshold of P<0.05.

[0035] In the present disclosure, it was evaluated both the ability of the studied drugs to correct or mitigate DNA damage and to change the levels of the CSB protein. In order to define whether the drugs were having a positive impact in reducing DNA damage, quantitative analysis was performed through comet assay taking into consideration three parameters: i) Tail DNA percent, ii) Olive moment and iii)Tail moment. In order to assess this information OpenComet software was used. Once those information’s were accessed, optimal cut off points were created through optimal binding, allowing to identify in specific interval of values where the majority of each condition was placed. Once the majority of each condition was scored with values lower than the control in all the three parameters the drug was considered beneficial to repair DNA damage. Among all the drugs that have being study 23 of than have present promising values as it can be seeing in table 2.

[0036] Table 2 - Representative data of (A) Tail DNA percent, (B) tail moment and (C) olive moment within optimal distribution. Values are shown in percentage, depicting the distribution of the majority of the values according with each optimal point, that was defined based on the change of the value’s normal distribution. A Tail DNA Percent 183

[0037] The drugs that presented a high number of values scoring lower than the control group, were selected for western blot analyses. To investigate the expression of CSB, total protein was extracted from cells that have been treated with the drugs, along with a control group. All the 23 drugs did not present a significant increase in the expression of CSB statistically confirmed by one-way ANOVA (P>0.05). (see figure 1 and 2).

[0038] Despite the 23 drugs that have shown significant result other 49 drugs that been tested in this study presented no significant different in any of the three different parameters (Tail moment, Olive Moment, Tail DNA %), showing no improvement in the correction of DNA damage. (Table 3) Table 3 – Statistical representation of all the drugs that have been tested taking into consideration tail moment, olive moment and tail DNA percent in which non presented statical significant difference, only 23 drugs of the 49 tested drugs have shown activity in the prevention or treatment of DNA repair-deficiency disorder caused by ageing related diseasesDrug Tail DNA percentage Tail Moment Olive MomentMeticrane >0.05 >0.05 >0.05Hydroflumethiazide >0.05 >0.05 >0.05Azaguanine-8 >0.05 >0.05 >0.05Allantoin >0.05 >0.05 >0.05Acetazolamide >0.05 >0.05 >0.05Diflunisal >0.05 >0.05 >0.05Procaine hydrochloride>0.05 >0.05 >0.05Moxisylyte hydrochoride>0.05 >0.05 >0.05Amyleine hydrochloride>0.05 >0.05 >0.05Lidocaine hydrochloride>0.05 >0.05 >0.05Ethosuximide >0.05 >0.05 >0.05Mafenide hydrochloride>0.05 >0.05 >0.05Nitrofurantoin >0.05 >0.05 >0.05Tranylcypromine hydrochloride>0.05 >0.05 >0.05Aceclofenac >0.05 >0.05 >0.05Sulfamethoxazole >0.05 >0.05 >0.05Mephenesin >0.05 >0.05 >0.05Phenformin hydrochloride>0.05 >0.05 >0.05Pargyline hydrochloride>0.05 >0.05 >0.05Pentolinium bitartrate >0.05 >0.05 >0.05Aminopurine, 6-benzyl >0.05 >0.05 >0.05Lomefloxacin hydrochloride>0.05 >0.05 >0.05Orphenadrine hydrochloride>0.05 >0.05 >0.05Dilazep dihydrochloride>0.05 >0.05 >0.05Drug Tail DNA percentage Tail Moment Olive MomentTolbutamide >0.05 >0.05 >0.05Nimesulide >0.05 >0.05 >0.05Salbutamol >0.05 >0.05 >0.05Prilocaine hydrochloride >0.05 >0.05 >0.05(S,+) Camptothecine >0.05 >0.05 >0.05Ranitidine hydrochloride >0.05 >0.05 >0.05Tiratricol >0.05 >0.05 >0.05Flufenamic acid >0.05 >0.05 >0.05Flumequine >0.05 >0.05 >0.05Tibolone >0.05 >0.05 >0.05Trimethoprim >0.05 >0.05 >0.05Piroxicam >0.05 >0.05 >0.05Gemfibrozil >0.05 >0.05 >0.05Mefexamide hydrochloride >0.05 >0.05 >0.05Fenbufen >0.05 >0.05 >0.05Ketoprofen >0.05 >0.05 >0.05Norfloxacin >0.05 >0.05 >0.05Antimycin A >0.05 >0.05 >0.05Nifenazone >0.05 >0.05 >0.05Griseofulvin >0.05 >0.05 >0.05Phentolamine hydrochloride >0.05 >0.05 >0.05Etodolac >0.05 >0.05 >0.05Scopolamin-N-oxide hydrobromide >0.05 >0.05 >0.05

[0039] The goal of this the present disclosure was to identify known drugs from a with (a) the ability of a drug to reduce DNA damage and (b) its relation of the levels of CSB expression. Using the comet assay to access the DNA damage, it was found that among the studied drugs, the 23 tested drugs of the present disclosure (Hydralazine hydrochloride, Mafenide hydrochloride, Iproniazide phosphate, (S,+) Camptothecine, Tolfenamic acid, Fenspiride hydrochloride, Mebendazole, Tropicamide, Riluzole hydrochloride, Clemizole hydrochloride, Atracurium besylate, Sulfacetamide sodichydrate, Heptaminol hydrochloride, Sulfathiazole, Hydrochlorothiazide, Idoxuridine, Isoflupredone acetate, Amiloride hydrochloride dihydrate, Amprolium hydrochloride, Betazole hydrochloride, Isoxicam, Ticlopidine hydrochloride, Dicyclomine hydrochloride, Levodopa) significantly reduced the levels of damage on the DNA. These drugs had scored lower values in all the parameters that were analyzed. Within the optimal binding test was possible to place the majority of each condition values within an interval.

[0040] Three particular class of treatment were extensively present among the drugs that have shown a geroprotective effect, among them cardiovascular, infectiology and central nervous therapeutic class that together some 52% of all drugs found on the present disclosure (table 1) what could indicate a pattern. However, is worth to mention that the number of drugs among therapeutical class in the Prestwick library is not the same, varying based on each drug class. In this library four particular groups present the most dominance among the different therapeutic class, namely metabolic, cardiovascular, central nervous system and infectiology treatments, what could indicate that the pattern observed in this study was only based on the amount of each drug present on the library.

[0041] Another goal of the present disclosure were to identify drugs that impacted CSB levels. Through western blot was possible to see that all drugs - Hydralazine hydrochloride, Mafenide hydrochloride, Iproniazide phosphate, (S,+) Camptothecine, Tolfenamic acid, Fenspiride hydrochloride, Mebendazole, Tropicamide, Riluzole hydrochloride, Clemizole hydrochloride, Atracurium besylate, Sulfacetamide sodic hydrate, Heptaminol hydrochloride, Sulfathiazole, Hydrochlorothiazide, Idoxuridine, Isoflupredone acetate, Amiloride hydrochloride dihydrate, Amprolium hydrochloride, Betazole hydrochloride, Isoxicam, Ticlopidine hydrochloride, Dicyclomine hydrochloride, Levodopa- did interfere with the CSB expression level, by both increasing or decreasing its expression however none of them interfered significantly with it.

[0042] In the present disclosure it was possible to identify 23 drugs that presented a positive effect on the reduction of DNA damage. These drugs were tested to identify if they mechanism of action was CSB dependent or independent take into consideration increase on the expression level of this protein. However, none of the drugs present a significant increase in the expression of CSB indicating a possible therapeutical approach for the treatment of disease that have as its main caused the lack of CSB protein such as Cockayne Syndrome. Nevertheless, more importantly was the fact that those drugs p- Hydralazine hydrochloride, Mafenide hydrochloride, Iproniazide phosphate, (S,+) Camptothecine, Tolfenamic acid, Fenspiride hydrochloride, Mebendazole, Tropicamide, Riluzole hydrochloride, Clemizole hydrochloride, Atracurium besylate, Sulfacetamide sodic hydrate, Heptaminol hydrochloride, Sulfathiazole, Hydrochlorothiazide, Idoxuridine, Isoflupredone acetate, Amiloride hydrochloride dihydrate, Amprolium hydrochloride, Betazole hydrochloride, Isoxicam, Ticlopidine hydrochloride, Dicyclomine hydrochloride, Levodopa - resented the capacity to assist on the recovery of a hallmark of aging being able to reduce the damage present on the DNA, possibly reducing the formation of mutations that could lead to miss function of cell homeostasis.

[0043] The term "comprising" whenever used in this document is intended to indicate the presence of stated features, integers, steps, components, but not to preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.

[0044] Furthermore, where the claims recite a composition, it is to be understood that methods of using the composition for any of the purposes disclosed herein are included, and methods of making the composition according to any of the methods of making disclosed herein or other methods known in the art are included, unless otherwise indicated or unless it would be evident to one of ordinary skill in the art that a contradiction or inconsistency would arise.

[0045] The disclosure should not be seen in any way restricted to the embodiments described and a person with ordinary skill in the art will foresee many possibilities to modifications thereof.

[0046] The above-described embodiments are combinable.

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Claims

C L A I M S1. Compound for use in the prevention or treatment of DNA repair-deficiency disorder caused by ageing related diseases, disorders or symptoms, wherein the compound is selected from a list consisting of: Hydralazine hydrochloride, Mafenide hydrochloride, Iproniazide phosphate, (S,+) Camptothecine, Tolfenamic acid, Fenspiride hydrochloride, Mebendazole, Tropicamide, Riluzole hydrochloride, Clemizole hydrochloride, Atracurium besylate, Sulfacetamide sodic hydrate, Heptaminol hydrochloride, Sulfathiazole, Hydrochlorothiazide, Idoxuridine, Isoflupredone acetate, Amiloride hydrochloride dihydrate, Amprolium hydrochloride, Betazole hydrochloride, Isoxicam, Ticlopidine hydrochloride, Dicyclomine hydrochloride, Levodopa and combinations thereof.

2. Compound for use according to the previous claim, wherein said use is for the prevention or treatment of DNA repair-deficiency disorder, disease, or symptoms caused by ageing related diseases, disorders or symptoms from the group comprising: Cockayne Syndrome type A, Cockayne Syndrome Type B, Xeroderma pigmentosum, Hutchinson-Gilford progeria syndrome.

3. Compound for use according to any the previous claims for reducing DNA damage associated with ageing-related pathologies and / or CSB mutations.

4. Compound for use according to any the previous claims for enhancing DNA repair mechanisms influenced by mutations in the CSB gene, CSA gene, XPC gene, XPA gene, XPB gene, TFIIH gene; preferably CSB gene.

5. Compound for use according to the previous claim, wherein the gene is CSB gene, CSA gene; preferably CSB gene.

6. Compound for use according to any of the previous claims, for use in the prevention or treatment of Premature Aging symptoms, UV Sensitivity Disorders, Neurological Disorders, Cockayne syndrome.

7. Pharmaceutical composition for use in the prevention or treatment of DNA repair- deficiency disorder caused by ageing related diseases, disorders or symptoms, comprising at least one compound according to any of the present claims in a therapeutical amount and a pharmaceutically acceptable carrier, adjuvant, excipient, emulsification agent or mixtures thereof.

8. Composition according to the previous claim, for use in the treatment of ageing related diseases, comprising for use in the treatment of ageing related diseases, comprising at least one compound according to any of the previous claim in a therapeutical amount and a pharmaceutically acceptable carrier, adjuvant, excipient, emulsification agent or mixtures thereof.

9. The use a compound for the manufacture of a medicament for the treatment of prevention or treatment of DNA repair-deficiency disorder caused by ageing related diseases, disorders or symptoms; wherein the compound is at least a compound according to any of the previous claims 1 – 6.

10. A method for treating or preventing DNA repair-deficiency disorder caused by ageing related diseases, disorders or symptoms in a subject, wherein the method comprising administering at least a compound according to any of the previous claims 1 – 6 to the subject.

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