N-alkyl-2-substituted ATP analogs for antibacterial agents

Abstract

The present invention relates to a compound of formula (I): TIFF2025521211000017.tif47170(wherein R l , R 2 , R 3 and X have the same meanings as defined in the claims and the specification), its tautomers, enantiomers or diastereomers. The present invention also relates to the use of such compounds as inhibitors of bacterial adhesion and biofilm formation on surfaces, an ex-vivo method for preventing the growth of bacteria in biofilm formation, and the use of such compounds in the diagnosis or prognosis of bacterial infections.

Description

Technical Field

[0001] The present invention relates to the newly discovered antibacterial properties of N-alkyl-2-substituted ATP analogs that are clinically used for another indication. The newly discovered antibacterial properties of the analogs can be applied to the prevention or treatment of bacterial infections in a subject, and further, the N-alkyl-2-substituted ATP analogs can be used ex-vivo or in vitro and can inhibit the adhesion of bacteria on the surface and biofilm formation.

Background Art

[0002] Cangrelor is a synthetic analog of adenosine triphosphate (ATP) belonging to N-alkyl-2-substituted ATP analogs and is a potent antagonist of the P2Y 12 receptor, which is a G protein-coupled purine receptor and an important component of platelet activation. Cangrelor is represented by the following formula II: TIFF2025521211000002.tif67170 or by its sodium salt represented by formula III: TIFF2025521211000003.tif69170.

[0003] Cangrelor is currently the only intravenous platelet P2Y 12 inhibitor available for clinical use. Cangrelor provides a rapid, potent and reliable antiplatelet effect. Due to such pharmacological properties, it can overcome the limitations of oral P2Y 12 inhibitors, which are characterized by an inevitable delay in onset of action that is enhanced in short-term situations with a higher risk of further impaired gastrointestinal absorption (Non-Patent Document 1).

[0004] Antibiotics are used for the prevention or treatment of bacterial infections. However, due to the misuse of antibiotics and the ability of bacteria to rapidly develop resistance mechanisms, we are currently facing a global spread of resistant bacteria worldwide. The currently available antibiotics are becoming increasingly ineffective, and treatment options for common infections such as urinary tract infections, sepsis, sexually transmitted infections, and some forms of diarrhea are limited. This phenomenon also threatens the safety of surgery, chemotherapy, or other treatments.

[0005] The World Health Organization predicts that antibiotic resistance will cause 10 million deaths annually by 2050. A study in The Lancet (2022) shows that resistant bacteria were directly involved in 1.27 million deaths in 2019 (Non-Patent Document 2).

[0006] Infections caused by Gram-positive bacteria are a major public health burden not only in terms of morbidity and mortality but also in terms of increased expenditure on patient management and implementation of infection control measures. More specifically, Staphylococcus aureus and enterococci are established pathogens in the hospital environment, and their frequent multidrug resistance complicates treatment.

[0007] Staphylococcus aureus is an important and highly virulent pathogen that causes a wide range of clinical symptoms, from relatively benign skin infections to life-threatening conditions such as endocarditis and osteomyelitis. It is also a commensal bacterium (colonizing approximately 30% of the population).

[0008] Two major shifts in S. aureus epidemiology have occurred since the 1990s: an epidemic of community-acquired skin and soft tissue infections (mainly driven by certain methicillin-resistant S. aureus [MRSA] strains), and an increase in the number of healthcare-associated infections (especially infective endocarditis and prosthetic device infections). Patients with MRSA infections are 64% more likely to die than patients with drug-susceptible infections.

[0009] Therefore, in this technical field, a new alternative to antibacterial therapy is urgently needed.

[0010] The pathogenicity of S. aureus depends on the expression of various pathogenicity factors. Among these factors, toxins are major pathogenicity determinants with cytotoxic properties that enable the bacteria to evade the host immune system. The carotenoid pigment staphyloxanthin helps S. aureus resist killing by host neutrophils via reactive oxygen species-dependent mechanisms. Bacterial pathogenicity is also determined by the ability of the bacteria to adhere to host tissues or surfaces through various mechanisms that are the first steps in bacterial colony formation, biofilm formation, and infection.

[0011] Antibacterial therapies that replace conventional antibiotics include agents that can inhibit bacterial pathogenicity factors, so-called anti-pathogenic drugs. Clinical trials are currently testing such anti-pathogenic drugs for the prevention or treatment of multi-drug resistant bacterial infections (Non-Patent Document 3). These anti-pathogenic drugs are valuable alternatives to antibiotics (Non-Patent Document 4). In fact, in contrast to antibiotics, these drugs do not impose a high selective pressure on bacteria, thus limiting the development of resistance and the spread of pathogenicity genes.

[0012] In 2017, C. Oury et al. described in Patent Document 1 a new use of triazolo[4,5-d]pyrimidine derivatives for the prevention and treatment of bacterial infections. However, triazolo[4,5-d]pyrimidine derivatives such as Ticagrelor are generally not soluble in an aqueous environment or medium.

Prior Art Documents

Patent Documents

[0013]

Patent Document 1

Non-Patent Documents

[0014]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Summary of the Invention

[0015] The present invention is based on the unexpected finding that at least one of the above objects can be achieved by N-alkyl-2-substituted ATP analogs that are clinically used for another indication.

[0016] The present invention provides N-alkyl-2-substituted ATP analogs having antibacterial properties. The analogs can inhibit the hemolytic activity of clinically relevant Staphylococcus aureus strains, such as clinical isolates from patients with infective endocarditis or methicillin-resistant strains. Furthermore, the inventors have observed that the analogs of the present invention can inhibit the production of staphyloxanthin, a major bacterial pigment. Since staphyloxanthin has antioxidant activity that helps bacteria avoid killing by reactive oxygen species produced by phagocytic innate immune cells, these data further indicate that the analogs of the present invention can reduce the pathogenicity of Staphylococcus aureus. Furthermore, the inventors have observed that the analogs of the present invention can reduce bacterial adhesion. Therefore, the compounds of the present invention are suitable for use as antibacterial agents, more particularly for the treatment and / or prevention of diseases caused by bacterial infections. As used herein, antibacterial agents also mean anti-pathogenic or anti-infective agents.

[0017] In contrast to triazolo(4,5-D)pyrimidine derivatives such as ticagrelol, the compounds of the present invention are very soluble in an aqueous environment or medium. This property enables administration by the intravenous route, leading to a very rapid onset of action, which can be important particularly in short-term situations where there is a high risk of impaired gastrointestinal absorption. The water-soluble property may also be advantageous for using the compounds of the present invention in aqueous compositions or coatings, such as hydrogels or nanogels or nanodispersions.

[0018] A first aspect of the present invention is of formula I for use in the prevention or treatment of bacterial infection in a subject: TIFF2025521211000004.tif69170(wherein, R l and R 2 are each independently hydrogen or halogen, R 3 and R 4 are each independently phenyl or C 1~6 alkyl, and each phenyl or C 1~6 alkyl is OR5 and C 1~6 alkylthio, NR 6 R 7 , phenyl, COOR 8 It can be substituted with one or more substituents independently selected from the group consisting of or halogen, or may be unsubstituted, R 5 , R 6 , R 7 and R 8 are each independently hydrogen or C 1~6 alkyl, X is an acidic moiety), a tautomer, enantiomer or diastereomer thereof, or a salt or solvate thereof, or a solvate of such a salt.

[0019] The present invention also relates to a pharmaceutical composition comprising a compound of formula (I) for use in the prevention or treatment of bacterial infection in a subject.

[0020] A further related aspect of the present invention provides the use of a compound of formula I described herein as an antibacterial agent, more specifically as an inhibitor of surface bacterial attachment and biofilm formation. In some embodiments, this use is a non-therapeutic use.

[0021] A further related aspect of the present invention provides an ex-vivo method for preventing the growth of bacteria in biofilm formation, comprising applying or grafting an effective amount of a compound of formula I described herein to a surface.

[0022] Yet a further related aspect of the present invention provides a compound of formula I described herein for use in the diagnosis or prognosis of bacterial infection. Such diagnosis or prognosis may be, for example, using a detectable marker or using any suitable method.

[0023] The following description of the figures of specific embodiments of the present invention is essentially merely illustrative and is not intended to limit the teachings of the present invention, their application or use.

Brief Description of the Drawings

[0024]

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BEST MODE FOR CARRYING OUT THE INVENTION

[0025] Before describing the present invention, it should be understood that the present invention is not limited to the specific processes, methods, and compounds described, and such processes, methods, and compounds can, of course, vary. Also, since the scope of the present invention is limited only by the appended claims, it should be understood that the terminology used herein is not intended to be limiting.

[0026] When describing the compounds and processes of the present invention, the terms used are construed in accordance with the following definitions, unless otherwise indicated by the context.

[0027] As used in the specification and the appended claims, the singular forms "a", "an", and "the" include both singular and plural referents unless the context clearly dictates otherwise. By way of example, "a compound" means one compound or a plurality of compounds.

[0028] As used herein, the terms "comprising", "comprises", and "comprised of" are synonymous with "including", "includes", or "containing", "contains", and are inclusive or open-ended and do not exclude additional, unrecited members, elements, or method steps. The terms "comprising", "comprises", and "comprised of" also include the term "consisting of".

[0029] As used herein, the term "about", when referring to a measurable value such as a parameter, quantity, duration, etc., means that such variations, as long as they are appropriate for carrying out the disclosed invention, include variations of not more than ±10%, preferably not more than ±5%, more preferably not more than ±1%, still more preferably not more than ±0.1% of the specific value. It should be understood that the value to which the modifying phrase "about" refers is itself also specifically, preferably, disclosed.

[0030] As used herein, the term "and / or", when used in a list of two or more items, means that any one of the listed items can be used by itself, or any combination of two or more of the listed items can be used. For example, if the list is described as including group A, group B, and / or group C, the list can include A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C.

[0031] The recitation of numerical ranges by endpoints includes all integers and, as appropriate, fractions included within that range (e.g., 1 to 5 can include 1, 2, 3, 4 when referring to a number of elements, for example, and can also include 1.5, 2, 2.75, and 3.80 when referring to measured values, for example). The recitation of endpoints includes the values of the endpoints themselves (e.g., 1.0 to 5.0 includes both 1.0 and 5.0). Any numerical range recited herein is intended to include all sub-ranges subsumed therein.

[0032] Throughout this specification, the reference to "one embodiment" or "an embodiment" means that a particular property, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment, although they may. Further, the particular properties, structures, or characteristics may be combined in any suitable manner in one or more embodiments, as will be apparent to those skilled in the art from this disclosure. Additionally, some embodiments described herein include some properties included in other embodiments but not others, and combinations of features of different embodiments are within the scope of the present invention and form different embodiments that can be understood by those skilled in the art. For example, in the following claims, any combination of the embodiments recited in the claims can be used.

[0033] Unless otherwise defined, all terms used in the disclosure of the present invention, including technical and scientific terms, have the meaning commonly understood by one of ordinary skill in the technical field to which the present invention pertains. Definitions of terms used in the description are included for better understanding of the teachings of the present invention.

[0034] When describing the present invention, the terms used are to be construed in accordance with the following definitions, unless otherwise indicated by the context.

[0035] The above terms and other terms used in this specification are well understood by those skilled in the art.

[0036] Whenever the term "substituted" is used in this specification, it means that one or more hydrogen atoms on the atom indicated by the expression using "substituted" are replaced by selection from the indicated groups, provided that the number of hydrogen atoms does not exceed the normal valence of the indicated atom and the substitution results in a chemically stable compound, i.e., a compound that is sufficiently robust to withstand isolation from the reaction mixture.

[0037] When the group can be substituted, such a group may be substituted with one or more, preferably one, two or three substituents. Preferred substituents include, but are not limited to, for example, halo, hydroxyl, alkyl, alkoxy, trifluoromethyl, trifluoromethoxy, cycloalkyl, aryl, arylalkyl, heterocyclyl, heteroaryl, cyano, amino, nitro, carboxyl, and monoalkylamino or dialkylamino, and can be selected from the group consisting of them.

[0038] The term "halo" or "halogen" as a group or part of a group is a general term for fluoro, chloro, bromo, and iodo.

[0039] The term "alkyl" refers to a hydrocarbyl group of the formula -C n H 2n+1 (wherein n is a number of 1 or more) as a group or part of a group. The alkyl group may be linear or branched and may be substituted as shown herein. Generally, the alkyl groups of the present invention contain from 1 to 6 carbon atoms, preferably from 1 to 5 carbon atoms, preferably from 1 to 4 carbon atoms, more preferably from 1 to 3 carbon atoms, and even more preferably 1 or 2 carbon atoms. In the present specification, when a subscript is used following a carbon atom, the subscript refers to the number of carbon atoms that the named group may contain. For example, "C 1~6 alkyl" includes all linear or branched alkyl groups having from 1 to 6 carbon atoms, and thus includes methyl, ethyl, n-propyl, i-propyl, butyl and its isomers (for example, n-butyl, i-butyl and t-butyl), pentyl and its isomers, hexyl and its isomers. For example, "C 1~5 alkyl" includes all linear or branched alkyl groups having from 1 to 5 carbon atoms, and thus includes methyl, ethyl, n-propyl, i-propyl, butyl and its isomers (for example, n-butyl, i-butyl and t-butyl), pentyl and its isomers. For example, "C 1~4"Alkyl" includes all linear or branched alkyl groups having 1 to 4 carbon atoms, and thus includes methyl, ethyl, n-propyl, i-propyl, butyl and its isomers (e.g., n-butyl, i-butyl and t-butyl). For example, "C 1~3 alkyl" includes all linear or branched alkyl groups having 1 to 3 carbon atoms, and thus includes methyl, ethyl, n-propyl, i-propyl.

[0040] The term "alkoxy" or "alkyloxy" refers to a group having the formula -OR b (wherein R b is alkyl as defined above herein). Non-limiting examples of suitable alkoxys include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy and hexyloxy.

[0041] "C 1~6 alkylthio" refers to a group having the formula -SR b (wherein R b is alkyl as defined above herein). Non-limiting examples of suitable C 1~6 alkylthios include methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, tert-butylthio, pentylthio and hexylthio.

[0042] The term "monoalkylamino or dialkylamino" refers to a group having the formula -N(R o )(R p (wherein R o and R p are each independently selected from hydrogen or alkyl, and at least one of R o or R p is alkyl). Thus, alkylamino includes monoalkylamino groups (e.g., mono C such as methylamino and ethylamino1~6 (alkylamino group), and dialkylamino groups (such as di-C such as dimethylamino and diethylamino 1~6 (alkylamino group). Non-limiting examples of suitable monoalkylamino or dialkylamino groups include n-propylamino, isopropylamino, n-butylamino, i-butylamino, sec-butylamino, t-butylamino, pentylamino, n-hexylamino, di-n-propylamino, di-i-propylamino, ethylmethylamino, methyl-n-propylamino, methyl-i-propylamino, n-butylmethylamino, i-butylmethylamino, t-butylmethylamino, ethyl-n-propylamino, ethyl-i-propylamino, n-butylethylamino, i-butylethylamino, t-butylethylamino, di-n-butylamino, di-i-butylamino, methylpentylamino, methylhexylamino, ethylpentylamino, ethylhexylamino, propylpentylamino, propylhexylamino, and the like.

[0043] Structural isomers are a type of isomer in which molecules with the same molecular formula have different bonding patterns and atomic organizations. When structural isomers can interconvert via a low-energy barrier, tautomeric isomerism (''tautomerism'') may occur. This can take the form of, for example, proton tautomerism in the compounds of the present invention containing an imino group, a keto group, or an oxime group, or so-called valence tautomerism in compounds containing an aromatic moiety. The compounds of formula I may exhibit tautomerism, such as imine-enamine tautomerism at the 6-position of adenine. Further, the compounds contain one or more asymmetric carbon atoms and thus exhibit optical and / or diastereoisomerism.

[0044] The present invention encompasses all possible stereoisomeric compounds of formula I and any subgroups thereof. If a compound is desired as a single enantiomer, such may be obtained by stereospecific synthesis, resolution of the final product or any convenient intermediate, or by chiral chromatography methods each known in the art. Resolution of the final product, intermediate, or starting material may be effected by any suitable method known in the art. See, for example, Stereochemistry of Organic Compounds by E. L. Eliel, S. H. Wilen, and L. N. Mander (Wiley-Interscience, 1994), which is incorporated herein by reference with respect to stereochemistry.

[0045] Preferred descriptions (features) and embodiments of the compounds and processes of the present invention are described below. Each description and embodiment of the invention so defined may be combined with any other description and / or embodiment, unless a separate express designation to the contrary is made. In particular, any feature shown as being preferred or advantageous may be combined with any other feature (s) shown as being preferred or advantageous.

[0046] A first aspect of the present invention is for use in the prevention or treatment of bacterial infection in a subject of formula I: TIFF2025521211000005.tif70170(wherein, R l and R 2 are each independently hydrogen or halogen, R 3 and R 4 are each independently phenyl or C 1~6 alkyl, each phenyl or C 1~6 alkyl being OR 5 , C 1~6 alkylthio, NR 6 R 7 , phenyl, COOR 8It can be substituted with one or more substituents independently selected from the group consisting of or halogen, or may be unsubstituted. R 5 , R 6 , R 7 and R 8 are each independently hydrogen or C 1~6 alkyl, X is an acidic moiety), a compound thereof, a tautomer, an enantiomer or a diastereomer or a salt or solvate thereof, or a solvate of such a salt is provided.

[0047] The present invention also relates to a pharmaceutical composition comprising a compound of formula I for use in the prevention or treatment of bacterial infections in a subject.

[0048] The pharmaceutical composition may contain, in addition to the compound of formula I, adjuvants, preservatives, solvents and / or viscosity modifiers. Solvents mean, for example, water, aqueous saline solution or any other physiological solution, ethanol, glycerol, oils such as vegetable oils or mixtures thereof. Viscosity modifiers mean, for example, carboxymethylcellulose and the like.

[0049] A further related aspect of the present invention provides the use of a compound of formula I described herein as an inhibitor of bacterial adhesion and biofilm formation on surfaces. In some embodiments, this use is a non-therapeutic use.

[0050] A further aspect of the present invention provides an ex-vivo method for preventing the growth of bacteria in biofilm formation, which comprises applying or grafting an effective amount of a compound of formula I described herein onto a surface.

[0051] An even further related aspect of the present invention provides a compound of formula I described herein for use in the diagnosis or prognosis determination of bacterial infections.

[0052] For example, when used for diagnosis or prognosis determination, the compound of formula I may contain a detectable marker. In some embodiments, the compound of formula I contains an isotope. In some embodiments, the isotope is 18 F.

[0053] As used herein, the term "detectable marker" refers to any type of tag that is detectable and thus enables determination of the presence of the compound of formula I. In certain embodiments, the marker is an isotope that enables use of the compound of formula I as a radioactive tracer. In some embodiments, the isotope is 18 F.

[0054] The acidic moiety, which may be represented by X, includes a Bronsted-Lowry acid, i.e., a moiety that acts as a proton donor. The acidic moiety can be monoacidic or polyacidic. In some embodiments, X is selected from the group consisting of -P(O)(OH)2, -SO3H, or -CO2H. In some embodiments, X is -P(O)(OH)2.

[0055] In some embodiments, R 4 is C 1~6 alkyl, and the C 1~6 alkyl is substituted with one, two, or three substituents. In some embodiments, R 4 is C 1~6 alkyl, and the C 1~6 alkyl is substituted with one, two, or three substituents independently selected from halo. In some embodiments, R 4 is C 1~6 alkyl, and the C 1~6 alkyl is substituted with two or three substituents selected from halogen, and the two or three substituents are the same.

[0056] In some embodiments, R 3 is C 1~6 alkyl, and the C 1~6The alkyl is substituted with one, two or three substituents. In some embodiments, R 3 is C 1~6 alkyl, and the C 1~6 alkyl is substituted with one, two or three substituents independently selected from C 1~6 alkylthio. In some embodiments, R 3 is C 1~6 alkyl, and the C 1~6 alkyl is substituted with one substituent selected from C 1~6 alkylthio. In some embodiments, R 3 is C 1~2 alkyl, and the C 1~2 alkyl is substituted with one substituent selected from methylthio.

[0057] In some embodiments, R l and R 2 are the same. In some embodiments, R l and R 2 are the same and represent Cl.

[0058] The compounds of the present invention may be in the form of salts as generally described below. Some preferred but non-limiting examples of suitable organic and / or inorganic acids are hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, acetic acid and citric acid, and other pharmaceutically acceptable acids known per se (see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002), which is incorporated herein by reference).

[0059] When the compounds of the present invention contain an acidic group in addition to a basic group, the compounds of the present invention can also form internal salts, and such compounds are within the scope of the present invention. When the compounds of the present invention contain a hydrogen-donating heteroatom (e.g., NH), the present invention also encompasses salts and / or isomers formed by transferring the above hydrogen atom to a basic group or atom within the molecule.

[0060] Salts of the compounds of formula I can be formed by reacting the free acid or its salt, or the free base, or its salt or derivative, with one or more equivalents of a suitable base or acid. The reaction can be carried out in a solvent or medium in which the salt is insoluble, or in a solvent in which the salt is soluble, such as ethanol, tetrahydrofuran or diethyl ether, and these solvents or media may be removed in vacuo or by lyophilization or other methods known in the art. The reaction may also be a metathetical process or may be carried out on an ion exchange resin.

[0061] The salts of the compound of formula I include its acid addition salts and basic salts. Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / monohydrogen phosphate / dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinafoate. Suitable basic salts are formed from bases that form non-toxic salts. Examples include aluminum salts, arginine salts, benzathine salts, calcium salts, choline salts, diethylamine salts, diolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, olamine salts, potassium salts, sodium salts, tromethamine salts, and zinc salts. Half salts of acids and bases, such as hemisulfate and hemicalcium salts, can also be formed. For a general review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002), which is incorporated herein by reference.

[0062] Examples of salts of the compound of formula I include alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts; salts of Group III elements such as aluminum salts; and ammonium salts. Examples of salts with suitable organic bases include salts with hydroxylamine; salts with lower alkylamines such as methylamine or ethylamine; salts with substituted lower alkylamines such as hydroxy-substituted alkylamines; or salts with monocyclic nitrogen heterocyclic compounds such as piperidine or morpholine; and salts with amino acids such as arginine, lysine, etc. or their N-alkyl derivatives; or salts with amino sugars such as N-methyl-D-glucamine or glucosamine. In some embodiments, the salt is a pharmaceutically acceptable salt. Pharmaceutically acceptable salts constitute non-toxic physiologically acceptable salts of the compound of formula I as described above herein.

[0063] In some embodiments, the compound is a salt, preferably the sodium salt of the compound of formula I.

[0064] In some embodiments, the compound of formula I is TIFF2025521211000006.tif68170, also known as tetrasodium cannabidiol or cannabidiol tetrasodium.

[0065] One aspect of the present invention provides a compound according to formula I described herein for use in the prevention or treatment of bacterial infection in a subject. In some embodiments, the bacterial infection is an infection caused by Gram-positive bacteria or Gram-negative bacteria.

[0066] Accordingly, the present invention also provides a method for treating bacterial infection in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound according to formula I described herein.

[0067] In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is a human host. In certain embodiments, the infection is a surface infection. The body surface includes, but is not limited to, the epidermis and mucous membranes. In some embodiments, the surface is a body surface. In some embodiments, the body surface is selected from an intact epidermal surface, a damaged epidermal surface, or a mucosal surface.

[0068] The damaged epidermal surface can be skin that is blistered, burned by fire, inflamed, pustular, sunburned, bitten, stabbed, or otherwise wounded. Suitable examples of mucosal surfaces include the mucosae of the mouth (including the tongue), nose, eyes, throat, esophagus, stomach, vagina, and rectum.

[0069] In some embodiments, the bacterial infection is an infection caused by Gram-positive bacteria. Examples of Gram-positive bacteria are Staphylococcus aureus, Staphylococcus epidermidis, Bacillus anthracis, Corynebacterium diphtheriae, Enterococcus faecalis, Enterococcus faecium, Erysipelothrix rhusiopathiae, Listeria monocytogenes, the genus Nocardia, Streptococcus pneumoniae, Streptococcus agalactiae. In some embodiments, the bacterial infection is an infection caused by Staphylococcus, preferably an infection caused by Staphylococcus aureus, preferably an infection caused by methicillin-resistant Staphylococcus aureus.

[0070] In some embodiments, the bacterial infection is caused by Gram-negative bacteria such as those of the genus Acinetobacter, for example, Acinetobacter baumannii, Bordetella pertussis, Campylobacter spp.; Enterobacteriaceae, for example, Citrobacter spp., Enterobacter spp., Escherichia coli, Klebsiella spp., Salmonella spp., Serratia marcescens, Shigella spp., Yersinia spp.; Haemophilus influenza, Helicobacter pylori, legionella pneumophila, Neisseria spp., Pseudomonas aeruginosa, Vibrio cholera, etc.

[0071] In some embodiments where the bacterial infection is caused by Gram-negative bacteria, the compounds of formula I described herein are used in combination with a membrane permeabilizing agent. In some embodiments, the membrane permeabilizing agent is selected from the group consisting of polymyxin B and polymyxin E.

[0072] A further related aspect of the invention provides for the use of the compounds of formula I described herein as inhibitors of bacterial adhesion and biofilm formation on surfaces. In some embodiments, this use is a non-therapeutic use.

[0073] The surface refers to not only any type of surface such as a rubber or plastic surface, for example, a surface made of polyethylene, polypropylene, polyurethane, polyvinyl chloride, polyvinyl pyrrolidone, polytetrafluoroethylene, silicone, or the like, or a copolymer, but preferably also a metal surface such as stainless steel, silver, gold, titanium, metal alloy, pyrolytic carbon, etc. In some embodiments, the surface is the surface of a biomaterial of a medical device. The present invention can also be used on the surface of a bioabsorbable or biomaterial such as a biological prosthesis or biological device made of a biological material such as porcine or bovine pericardium.

[0074] Staphylococcus aureus and coagulase-negative staphylococci (often Staphylococcus epidermidis) cause 65% to 75% of generator pocket infections and up to 89% of device-related endocarditis. Episodes occurring within two weeks after transplantation are likely to be caused by S. aureus.

[0075] Prosthetic valve endocarditis (PVE) is a serious infectious disease that can lead to fatal consequences. Bacteria can reach the prosthetic valve either by direct contamination during surgery or via hematogenous spread during the first few days and weeks after surgery. Bacteria have direct access to the perivalvular tissue along the interface between the prosthesis and the annulus and the suture pathway because the valve suture ring, cardiac annulus, and fixation sutures are not endothelialized early after valve implantation. These structures are coated with host proteins such as fibronectin and fibrinogen, and some organisms can attach to these and initiate infection.

[0076] The risk of developing prosthetic valve endocarditis (PVE) is highest in the first 3 months after surgery, remains high until 6 months, and then gradually declines to approximately 0.4% per year after 12 months post - surgery. The percentage of patients who develop PVE within the first year after valve replacement ranges from 1% to 3% in studies with active follow - up, and over 5 years, the cumulative percentage ranges from 3% to 6%.

[0077] The pathogens most frequently encountered in early PVE (within 2 months after implantation) are S. aureus and coagulase - negative staphylococci. The pathogens most frequently encountered in late PVE (after 2 months post - valve implantation) are streptococci and S. aureus, followed by coagulase - negative staphylococci and enterococci. Coagulase - negative staphylococci that cause PVE during the first year after surgery are almost exclusively Staphylococcus epidermidis. 84% to 87% of these organisms are methicillin - resistant and thus resistant to all beta - lactam antibiotics.

[0078] PVE is known to account for about 20% of all infective endocarditis. PVE is associated with health care in about 30% of cases. S. aureus is the leading causative pathogen and is involved in more than 20% of PVE.

[0079] Periprosthetic joint infection (PJI) occurs in 1% to 2% of joint replacements and is a major cause of failure of arthroplasty. Biofilms play an important role in the etiology of PJI. Bacteria within biofilms become resistant to treatment, and as a result, antimicrobial therapy often fails unless the biofilm is physically disrupted or removed by surgical debridement. Management of PJI generally consists of both surgery and antimicrobial therapy.

[0080] Accordingly, the use of the compounds of formula I according to the present invention is advantageous in the treatment, prevention and management of the above-mentioned conditions by administering the compounds of formula I to the surface of a medical device.

[0081] Medical devices include, but are not limited to, any device, tool, instrument, implant, etc. related to the practice of medicine or human or veterinary medicine, or intended for use in curing or treating a disease or condition. Medical devices can include all natural and synthetic materials, as well as both fibrous and non-fibrous materials. For example, the material may be composed of metal, plastic, paper, glass, ceramic, fiber, rubber, polymer, composite material, or any other material or combination of materials. Exemplary medical devices include, but are not limited to, any type of catheter, cannula; needle; stent of any size, shape, or configuration; coil of any size, shape, or configuration; contact lens; IUD; peristaltic pump chamber; endotracheal tube; gastrointestinal nutrition tube; arteriovenous shunt; condom; membranes for artificial lungs and kidneys; gloves; pacemaker lead; wound dressing; metal pins, plates and screws; metal artificial hip joints; artificial knee joints, etc.

[0082] In some embodiments, the surface is the surface of a cardiovascular device or the surface of a catheter. In some embodiments, the surface is the surface of an artificial heart valve, pacemaker, defibrillator, cardiac ablation catheter, cardiovascular formation device, assistive artificial heart, or catheter.

[0083] Examples of cardiovascular devices suitable for the present invention include artificial heart valves, pacemakers, defibrillators, cardiac ablation catheters, cardiovascular formation devices, ventricular assist devices (mechanical pumps), etc.

[0084] As used herein, the term "biomaterial" refers to a substance that has been engineered to interact with a biological system for medical purposes. Biomaterials can be used in applications such as joint replacements, bone plates, intraocular lenses, bone cements, artificial ligaments and tendons, dental implants, vascular prostheses, heart valves, skin repair devices (such as wound dressings), cochlear replacements, contact lenses, breast implants, drug delivery mechanisms, sustainable materials, vascular grafts, stents, nerve conduits, surgical sutures, wound closure clips and staples, pins, fracture stabilization screws, and surgical meshes.

[0085] In certain embodiments, the use according to this aspect of the invention involves applying a compound of formula I as described herein to the surface. In certain embodiments, the application is ensured by spraying a composition comprising the compound of formula I onto the surface.

[0086] The compound of formula I according to the invention may be applied to the surface of the above-mentioned biomaterial or medical device in the form of a coating.

[0087] As used herein, a "coating" is a covering applied to a surface. The coating of the surface can be achieved by any method known in the art, including but not limited to spraying a composition comprising the compound of formula I or immersing the device (or its surface) in a composition comprising the compound of formula I.

[0088] Accordingly, the invention also provides a composition comprising a compound of formula I as described herein, and a solubilizing vehicle.

[0089] Suitable solubilizing vehicles for preparing the compositions according to the invention include water, aqueous solutions, alcohols, dimethyl sulfoxide and mixtures thereof. In some embodiments, the solubilizing vehicle is an aqueous solution. Suitable aqueous solutions for preparing the compositions according to the invention include culture media (such as tryptic soy broth), aqueous saline solutions (in particular, physiological saline solutions), physiological solutions (such as frog Ringer's solution, Krebs solution, Tyrode's solution, Ringer Locke solution, De Jaren solution or artificial cerebrospinal fluid) and buffer solutions. In a preferred embodiment, the composition is in the form of a nano-dispersion or a nano-gel. A nano-dispersion means a composition having at least one phase within the nano-size range. Preferably, the compound of formula I described herein is in the nano-size phase. A nano-gel means a polymer-based composition in which the polymer is cross-linked to form a network. Preferably, the compound of formula I described herein is encapsulated in such a network. Advantageously, the nano-dispersion or nano-gel composition enhances the stability of the compound of formula I.

[0090] In some embodiments, the composition comprises: not including TIFF2025521211000007.tif68170.

[0091] In certain embodiments, the composition is a sprayable composition. Accordingly, the present invention also provides, without limitation, a container equipped with a spraying device such as a spray can containing the composition according to the invention. In certain embodiments, the composition (and / or the container) further comprises a gas propellant.

[0092] The compound of formula I according to the invention may also be encapsulated in a polymer network or a gel-like structure as described, for example, by C. Oury in International Publication No. WO 2018 / 122318.

[0093] Furthermore, a further related aspect of the present invention provides a medical device coated with a compound of formula I or a composition comprising a compound of formula I described herein, preferably a cardiovascular device, most preferably a catheter, an artificial heart valve or a pacemaker.

[0094] A further related aspect of the present invention provides an ex-vivo method for preventing the growth of bacteria in biofilm formation, which comprises applying or grafting an effective amount of a compound of formula I described herein onto a surface.

[0095] Bacteria in biofilms produce extracellular polymeric substances (EPS) mainly composed of polysaccharides, nucleic acids (extracellular DNA) and proteins, which protect the bacteria from external threats including immune system components and antibacterial agents. Furthermore, since the bacteria in biofilms have reduced metabolism, they are less sensitive to antibiotics, which is due to the fact that most antibacterial drugs require a certain degree of cell activity to be effective. Another factor that enhances such resistance is that the presence of the EPS matrix barrier impairs the diffusion of antibacterial drugs throughout the biofilm.

[0096] It has been determined that biofilm formation on any surface occurs in different states or steps. The first step of biofilm formation is binding / attachment to the surface, which is stronger under shear stress conditions. The protein mainly responsible for this attachment is polysaccharide intercellular adhesin (PIA), by which bacteria bind to each other and also to the surface to create a biofilm. The second stage of biofilm formation is the development of community structure and ecosystem, which produces a mature biofilm. The final stage is separation from the surface, as a result of which it spreads to other places. A quorum sensing (QS) system that mediates intercellular communication is involved in all phases of biofilm formation.

[0097] As used herein, "prevention of bacterial growth in biofilm formation" refers to inhibition of biofilm formation at all stages of its formation, starting not only with prevention or inhibition of bacterial attachment on the surface in Step 1, but mainly with inhibition of bacterial growth, propagation, and microcolony formation on the surface in Step 2. Inhibition of biofilm formation during the maturation Step 3 and inhibition of bacterial dispersion from the matrix in the colony formation step are also contemplated in this definition. Also, bacteria may be killed at all steps of biofilm formation.

[0098] Therefore, the use of the compounds of formula I according to the invention is advantageous in inhibiting bacterial biofilm formation on surfaces.

[0099] In some embodiments, a method of controlling bacterial growth in biofilm formation on a surface comprises applying or grafting an effective amount of a compound of formula I described herein to the surface in a preventive step of reducing bacterial attachment and survival on a substrate, at a stage where a biofilm is already present, or even in a maturation step involving matrix formation where a more complex structure of the biofilm that protects bacteria as a barrier against conventional antibacterial agents is established.

[0100] In some embodiments, the method of preventing bacterial growth can also be applied to the surface of an experimental device that requires such an antibacterial or anti-infection treatment.

[0101] A further related aspect of the invention provides a compound of formula I described herein for use in the diagnosis or prognosis of a bacterial infection. Advantageously, the compound of formula I can bind to bacteria and at the same time bind to platelets. Therefore, the compound of formula I has dual properties that enable the diagnosis or prognosis of a bacterial infection. For example, the compound of formula I can bind to lipids present in the bacterial membrane by its alkyl chain. Advantageously, through this dual binding possibility, the compound of formula I accumulates at the site of infection and can thus act as a tracer.

[0102] The compounds of formula I according to the present invention may be prepared by methods known to those skilled in the art. In particular, the synthetic methods described in WO 94 / 18216 may be used.

Examples

[0103] The following examples are provided for the purpose of illustrating the present invention and should in no way be construed as limiting the scope of the present invention.

[0104] Bacterial strains and growth conditions The Staphylococcus aureus strains included previously characterized infective endocarditis (IE) clinical isolates (Liesenborghs L, Meyers S, Lox M, Criel M, Claes J, Peetermans M, Trenson S, Vande Velde G, Vanden Berghe P, Baatsen P, et al. Staphylococcus aureus endocarditis: distinct mechanisms of bacterial adhesion to damaged and inflamed heart valves. Eur Heart J 2019; 40:3248-3259) or JE2 (USA300 MRSA). Bacteria were grown in tryptic soy broth (TSB, Sigma) at 37°C with agitation (200 rpm). Prior to all experiments, one colony from a TSA plate was used to inoculate 4 mL of TSB in a 15 mL polystyrene tube and the bacteria were grown overnight. The liquid culture was then diluted 100-fold with 20 mL of fresh TSB and dispensed into new tubes. For the treatment with sodium tetracangrelol, various concentrations of the drug were added to the corresponding tubes. Sodium tetracangrelol was dissolved in the culture medium, i.e., tryptic soy broth containing proteins and nutrients for growing bacteria, more specifically, casein, soy peptone, sodium chloride, dipotassium phosphate, dextrose, water. The supernatant and bacterial pellet were collected at the designated time (logarithmic growth phase or stationary growth phase). The supernatant was filtered through a 0.22 μM cellulose acetate filter (VWR) and stored at -20°C until further use.

[0105] Labeling of bacteria Logarithmic-phase Staphylococcus aureus JE2 (USA300 MRSA) that was untreated or treated with 1 μg / ml of canagrelol was stained with 30 μg / ml of 5(6)-carboxyfluorescein N-hydroxysuccinimide ester (Sigma-Aldrich) in phosphate-buffered saline (PBS) for 30 minutes at room temperature on a shaker. After incubation, the bacteria were centrifuged at 14,000×g for 5 minutes and the supernatant was removed. Next, the bacteria were resuspended in PBS and the washing step was performed by removing the PBS after centrifugation (14,000×g for 5 minutes). Then, the bacterial pellet was resuspended in 100 μl of 6% bovine serum albumin (BSA). The stained bacteria were stored on ice until further used in von Willebrand factor (VWF) adhesion assay and fibrinogen (Fg) adhesion assay.

[0106] Preparation of erythrocyte suspension and hemolysis assay Citrate-anticoagulated blood freshly drawn from healthy volunteers was centrifuged at 1,000×g for 15 minutes. After discarding the plasma supernatant, the erythrocytes were washed once with phosphate-buffered saline (PBS) and resuspended at 2% in PBS. The supernatant from an overnight bacterial culture was mixed with the freshly prepared human erythrocyte suspension at a final ratio of 1:5 and incubated for 30 minutes with gentle agitation on a table shaker (800 rpm). Then, the samples were centrifuged at 4,000×g for 5 minutes. Hemolytic activity was determined in the supernatant by measuring the absorbance at 570 nm with a spectrophotometer.

[0107] Analysis of staphyloxanthin production Bacterial pellets from overnight cultures were resuspended in 400 μL of methanol and incubated at 37°C for 30 minutes with gentle agitation on a table shaker. The samples were centrifuged at 14,000×g for 5 minutes to remove cell debris. The pigment intensity in the supernatant was analyzed by measuring the absorbance at 470 nm.

[0108] Example 1: Sodium canagrelol inhibited the hemolytic activity of Staphylococcus aureus isolated from patients with infective endocarditis The supernatant was prepared from a Staphylococcus aureus IE clinical isolate grown to the logarithmic phase in the absence (control) or presence of sodium cangrelor (0.5 μg / mL) and added to the erythrocyte suspension for 30 minutes before assessing the hemolytic effect. Sodium cangrelor was purchased from Bio-Techne Ltd. with reference to the AR-C 69931 tetrasodium salt. The 0.5 μg / mL concentration of sodium cangrelor corresponds to the steady-state concentration of the drug achieved in patients receiving conventional antiplatelet administration, i.e., an intravenous bolus (30 μg / kg) followed by an infusion of 4 μg / kg / min (Akers, W.S., Oh, J.J., Oestreich, J.H., Ferraris, S., Wethington, M. and Steinhubl, S.R. Pharmacokinetics and Pharmacodynamics of a Bolus and Infusion of Cangrelor: A Direct, Parenteral P2Y12 Receptor Antagonist. The Journal of Clinical Pharmacology 2010; 50: 27-35). Figure 1 shows that when bacteria were grown in the presence of sodium cangrelor, the supernatant hemolytic activity was lost by 90% compared to the vehicle control (set at 100%). These data indicate that cangrelor can reduce the pathogenicity of clinical Staphylococcus aureus strains.

[0109] Example 2: Sodium cangrelor inhibited the hemolytic activity of methicillin-resistant Staphylococcus aureus The supernatant was prepared from a methicillin-resistant JE2 (USA300) Staphylococcus aureus strain grown to stationary phase in the absence (control) or presence of kanglerin (1 μg / mL) and added to the erythrocyte suspension for 30 minutes before assessing the hemolytic effect. Figure 2 shows that when bacteria were grown in the presence of kanglerin, the supernatant hemolytic activity was completely lost compared to the vehicle control (set at 100%). These data indicate that kanglerin can reduce the pathogenicity of methicillin-resistant Staphylococcus aureus strains.

[0110] Example 3: Tetrasodium kanglerin inhibited staphyloxanthin production by Staphylococcus aureus Protein extracts from Staphylococcus aureus IE clinical isolates grown to logarithmic phase in the absence (control) or presence of kanglerin (0.5 μg / mL) were used to measure the content of staphyloxanthin, the major bacterial pigment. Figures 3 and 4 show that when bacteria were grown in the presence of kanglerin, bacterial staphyloxanthin production was lost by 90% compared to the vehicle control (set at 100%). Since staphyloxanthin has antioxidant activity that helps bacteria avoid killing by reactive oxygen species produced by phagocytic innate immune cells, these data further indicate that kanglerin can reduce the pathogenicity of Staphylococcus aureus.

[0111] Example 4: von Willebrand factor adhesion experiment Cover glass (24 mm × 60 mm) was coated with 50 μg / ml VWF (Haemate P, CSL Behring, Mechelen, Belgium) at room temperature for 4 hours. The coated cover glass was attached to a flow chamber system (Provenzale, I., Brouns, S. L. N., van der Meijden, P. E. J., Swieringa, F. & Heemskerk, J. W. M. Whole blood based multiparameter assessment of thrombus formation in standard microfluidic devices to proxy in vivo haemostasis and thrombosis. Micromachines vol. 10 787 (2019)). Logarithmic phase labeled bacteria were diluted to approximately 10 7 CFU / ml to 10 8 CFU / ml in TSB medium. Using a flow chamber system, 1 ml of the bacterial mixture was perfused onto the VWF-coated surface using a 1 ml syringe. The cover glass was attached to the flow chamber, and the labeled bacteria were perfused at a shear rate of 1000 s -1 for 10 minutes with a high-precision Harvard pump (PHD 11 plus, Harvard Apparatus, Holliston, Massachusetts, USA).

[0112] After perfusion, the coverslip was removed from the system, washed with PBS, and then fixed for 10 minutes with 4% paraformaldehyde (PFA, miliporefore, Merck). The slides were mounted using ProLong™ Diamond Antifade Mountant (Invitrogen). Confocal microscopy was performed at 40× magnification, and fluorescence intensity was measured using QuPath software (Bankhead, P. et al. QuPath: Open source software for digital pathology image analysis. Sci. Rep. 7, 1-7 (2017)). Quantification of bacterial adhesion to VWF was determined by determining the ratio of the fluorescence signal to the proportion of positive pixels corresponding to bacterial adhesion. Figure 5 shows that growth of JE2 bacteria in the presence of 1 μg / ml of kanglerinol decreased bacterial adhesion to von Willebrand factor under flow compared to the vehicle control.

[0113] Example 5: Fibrinogen Adhesion Experiment A 96-well plate (cellstar, greiner) was coated with human plasma fibrinogen (Fg, Merck) at room temperature for 24 hours, followed by a washing step with PBS. Next, a blocking step with 2% BSA was performed for 1 hour, followed by a washing step with PBS. Then, the labeled bacteria in the logarithmic phase were diluted in TSB medium to 10 9Labeled bacteria were obtained at CFU / ml. Subsequently, the bacterial solution was incubated at 37 °C for 1 hour and washed with PBS. The adherent bacteria were fixed with 4% PFA for 10 minutes and washed again. Next, the attachment of bacteria was quantified using a spectrophotometer (Multimode Microplate Reader FilterMax F5, molecular devices) with excitation at 485 nm and emission at 535 nm, and a surface scan of the well was performed. The initial amount of bacteria was seeded on TSA plates to determine the exact CFU / ml. Finally, the fluorescence signal was scaled according to the obtained CFU / ml. Figure 6 shows that when JE2 bacteria were grown in the presence of 1 μg / ml of kanglerol, the attachment of bacteria to fibrinogen decreased compared to the vehicle control.

[0114] Example 6: Bacterial growth curve and metabolic activity The bacterial growth curve and metabolic activity profile of logarithmic-phase JE2 bacteria were obtained after treatment with various concentrations of kanglerol: 1 μg / ml, 20 μg / ml, and 40 μg / ml. The bacterial growth curve was obtained by kinetic readings using a spectrophotometer and was read every 10 minutes for 7 hours at an optical density of 595 nm with continuous shaking. The metabolic activity profile was obtained using calscreener (trademark) (Symcel), which measures the metabolic reaction over time (15 hours) in the presence of various concentrations of kanglerol. Figures 7 and 8 show that growing JE2 bacteria in the presence of various concentrations of kanglerol, including 1 μg / ml, 20 μg / ml, and 40 μg / ml, did not affect bacterial growth or bacterial metabolic activity compared to the vehicle control. Kanglerol does not inhibit the bacterial growth of JE2, but when treated with a clinically relevant dose of 1 μg / ml, it can inhibit the production of pathogenic factors by this strain.

[0115] Example 7: Detectable marker 18 As an N-alkyl-2-substituted ATP analog containing F 18 Preparation of F-kanglerol and bacterial uptake Using the strategy previously described by Josse et al. in Bioorganic and Medicinal Chemistry 9 (2001) 665 and by Cheguillaume et al. in Bioorganic and Medicinal Chemistry 13 (2005) 1357, synthesize a sulfur precursor for 18 F]-labeling by oxidative fluorodesulfurization reaction. Then, 18 prepare

[0116] Next, analyze the selective uptake of 18 F]-cangrelol into the bacterium S. epidermidis. Incubate the bacteria 18 with

[0117] F]-cangrelol and measure the relative radioactivity associated with the bacterial cells. 600 Grow S. epidermidis bacteria overnight at 37 °C in tryptic soy broth (TSB) with shaking at 250 rpm. Dilute the overnight culture to OD 8 0.1 and incubate until the mid-log phase is reached. Resuspend 1 × 10 18 CFU in 1 ml of cell culture medium RPMI 1640 (R7638) provided by Sigma-Aldrich. Incubate the bacteria and a bacteria-free control with 2 MBq of 2 F]-cangrelol at 37 °C for 1 h. Recover the bacteria by centrifugation (600 × g, 5 min) and wash three times by successive centrifugation. After washing, transfer the cells to a scintillation vial. Collect the supernatant also in the scintillation vial. Count the bacteria and the supernatant by a gamma counter (2470 Wizard

[0118] In the bacterial pellet 18 F]-cangrelol activity is observed, indicating the possibility of bacterial uptake and the usefulness of the tracer for the diagnosis of bacterial infections.

Claims

1. A compound of formula I: for use in the prevention or treatment of bacterial infections in a subject (wherein R ｌ and R ２ are each independently hydrogen or halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl can be substituted with one or more substituents each independently selected from the group consisting of OR ５ , C １～６ alkylthio, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted. R ５ 、 R ６ 、 R ７ and R ８ are each independently hydrogen or C １～６ alkyl, X is an acidic moiety), its tautomer, enantiomer or diastereomer, or a salt or solvate thereof, or a solvate of such a salt.

2. X is -P(O)(OH) ２ , -SO ３ H, or -CO ２ H, and is selected from the group consisting of, the compound according to claim 1

3. R ４ is C １～６ alkyl, and the C １～６ alkyl is substituted with one, two or three substituents, the compound according to claim 1 or 2.

4. R ３ is C １～６ alkyl, and said C １～６ alkyl is substituted with one, two or three substituents, the compound according to any one of claims 1 to 3.

5. R ｌ and R ２ The compound according to any one of claims 1 to 4, wherein they are the same.

6. The compound according to any one of claims 1 to 5, wherein the compound is a salt, preferably a sodium salt of the compound of formula I.

7. The compound according to any one of claims 1 to 6, which is tetrasodium cangrelor of formula III:

8. The compound according to any one of claims 1 to 7, wherein the bacterial infection is an infection caused by a Gram-positive bacterium, preferably an infection caused by a staphylococcus, preferably an infection caused by Staphylococcus aureus, preferably an infection caused by methicillin-resistant Staphylococcus aureus.

9. The compound according to any one of claims 1 to 8, wherein the bacterial infection is an infection on the body surface.

10. A compound of formula I: as an antibacterial agent (wherein R ｌ and R ２ are each independently hydrogen or a halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl may be substituted with one or more substituents each independently selected from the group consisting of OR ５ , thio C １～６ alkyl, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted. R ５ 、R ６ 、R ７ and R ８ are each independently hydrogen or C １～６ alkyl, X is an acidic moiety), its tautomer, enantiomer or diastereomer, or a salt or solvate thereof, or a solvate of such a salt for ex-vivo or in vitro use.

11. Use of a compound of formula I: as an inhibitor of bacterial adhesion and biofilm formation on a surface (wherein R ｌ and R ２ are each independently hydrogen or halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl can be substituted with one or more substituents each independently selected from the group consisting of OR ５ , C １～６ alkylthio, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted. R ５ 、 R ６ 、 R ７ and R ８ are each independently hydrogen or C １～６ alkyl, X is an acidic moiety), its tautomer, enantiomer or diastereomer, or a salt or solvate thereof, or a solvate of such a salt.

12. The use according to claim 11, wherein the surface is the surface of a biomaterial or a medical device, preferably the surface of a cardiovascular device or the surface of a catheter, most preferably the surface of a catheter, an artificial heart valve, or a pacemaker.

13. An ex-vivo method for preventing the growth of bacteria in biofilm formation, comprising applying or grafting an effective amount of a compound of formula I: (wherein R ｌ and R ２ each independently represents hydrogen or halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl can be substituted with one or more substituents each independently selected from the group consisting of OR ５ , C １～６ alkylthio, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted. R ５ 、 R ６ 、 R ７ and R ８ are each independently hydrogen or C １～６ alkyl, X is an acidic moiety), its tautomer, enantiomer or diastereomer, or a salt or solvate thereof, or a solvate of such a salt onto a surface.

14. A compound of formula I: for use in the diagnosis or prognosis determination of bacterial infections (wherein R ｌ and R ２ are each independently hydrogen or halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl may be substituted with one or more substituents each independently selected from the group consisting of OR ５ , C １～６ alkylthio, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted. R ５ 、 R ６ 、 R ７ and R ８ each independently is hydrogen or C １～６ alkyl, A compound in which X is an acidic moiety), its tautomer, enantiomer or diastereomer, or a salt or solvate thereof, or a solvate of such a salt. **Claim 15** A medical device, preferably a cardiovascular device, most preferably a catheter, an artificial heart valve or a pacemaker, which has the formula I: (wherein, R ｌ and R ２ are each independently hydrogen or a halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl may be substituted with one or more substituents each independently selected from the group consisting of OR ５ , C １～６ alkylthio, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted. R ５ 、R ６ 、R ７ and R ８ are each independently hydrogen or C １～６ alkyl, A compound in which X is an acidic moiety), its tautomer, enantiomer or diastereomer, or a medical device coated with a composition comprising a salt or solvate thereof, or a solvate of such a salt. **Claim 16** A pharmaceutical composition for use in the prevention or treatment of bacterial infection in a subject, having the formula (I): (wherein, R ｌ and R ２ are each independently hydrogen or a halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl can be substituted with one or more substituents each independently selected from the group consisting of OR ５ , C １～６ alkylthio, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted, R ５ 、 R ６ 、 R ７ and R ８ are each independently hydrogen or C １～６ alkyl, A compound in which X is an acidic moiety), its tautomer, enantiomer or diastereomer, or a salt or solvate thereof, or a solvate of such a salt. **Claim 17** A composition comprising: - a compound of formula I, its tautomer, enantiomer, or diastereomer, and - a solubilizing vehicle, wherein the compound of formula I is a compound having the structure: (wherein, R ｌ and R ２ are each independently hydrogen or halogen, R ３ and R ４ are each independently phenyl or C １～６ alkyl, and each phenyl or C １～６ alkyl can be substituted with one or more substituents each independently selected from the group consisting of OR ５ , C １～６ alkylthio, NR ６ R ７ , phenyl, COOR ８ or halogen, or may be unsubstituted. R ５ , R ６ , R ７ and R ８ are each independently hydrogen or C １～６ alkyl, X is an acidic moiety), or a salt or solvate thereof, or a solvate of such a salt.

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