Antibiotic compounds for treating bacterial infections associated with a staphylococcal species

Abstract

The present invention provides compounds of general Formula (I) which are suitable for treating bacterial infections associated with a Staphylococcal species.

Description

Antibiotic compounds for treating bacterial infections associated with a Staphylococcal speciesField of the Invention

[0001] The present invention provides compounds for use in a method of treating bacterial infections associated with a Staphylococcal species, in particular infections associated with Staphylococcus aureus. Moreover, the present invention provides a novel antibiotic compound that is useful for treating bacterial infections associated with a Staphylococcal species.Background of the Invention

[0002] In recent years, resistance to standard antibiotic agents has risen dramatically, especially in relation to bacterial infections associated with Staphylococcal infections and, in particular, infections associated with Staphylococcus aureus (5. aureus). For example, such resistant 5. aureus includes MRSA, resistant to methicillin, vancomycin, linezolid and many other classes of antibiotics, or the newly discovered New Delhi metallo-beta-lactamase-1 (NDM-1) type resistance that has shown to afford bacterial resistance to most known antibacterials, including penicillins, cephalosporins, carbapenems, quinolones and fluoroquinolones, macrolides, etc. This situation threatens our ability to effectively treat and prevent an ever-increasing range of infections. As a result, there is an ongoing need for the development of novel antibiotic compounds that are effective in cases where established antibiotics fail.

[0003] A recently developed new class of antibiotics compounds are Fabl inhibitors. These compounds inhibit the NADH-dependent enoyl reductase (Fabl) from the type II bacterial fatty acid biosynthesis pathway (FAS-II) and thus selectively affect bacteria relying on this enzyme without affecting the host (patient). Fabl inhibitors therefore provide an alternative approach for treating bacterial infections in cases where established antibiotics fail. Advantageously, this mode of action is not expected to display any cross-resistance to established antibiotics. For example, WO 2020 / 0993341 Al describes Fabl inhibitor compounds showing antibiotic activity against N. gonorrhoeae. However, these known Fabl inhibitor compounds are not described in the context of treating Staphylococcal infections. Furthermore, Fabl inhibitor compounds showing antibiotic activity against 5. aureusinfections are described in the state of the art. However, these compounds may not be well tolerated.

[0004] Accordingly, there is still a need for compounds and pharmaceutical compositions comprising the same which can show antibiotic activity (especially in cases where established antibiotics fail) against a Staphylococcal species, in particular against 5. aureus. It is preferable that such compounds do not give rise to cross-resistance to established antibiotics, and it is desirable that such compounds give rise to a low / acceptable rate of side effects.

[0005] It is therefore an object of the present invention to address one or more of the aforementioned needs. Further objectives and problems underlying the present invention may become apparent from the subsequent description.Summary of the Invention

[0006] The present invention is based on the discovery of compounds showing antibacterial activity against bacterial infections associated with a Staphylococcal species. In particular, the inventors have surprisingly found that the compounds claimed herein show excellent antibacterial activity against bacterial infections associated with a Staphylococcal species, in particular against infections associated with Staphylococcus aureus, while at the same time being well tolerated and less prone to give rise to undesirable side effects. The presently claimed compounds can therefore be used in the treatment of bacterial infections associated with a Staphylococcal species.

[0007] Thus, the present invention provides in particular the following:

[0008] 1. A compound for use in a method of treating a bacterial infection selected from infections associated with a Staphylococcal species in a patient, wherein the compound is represented by Formula IFormula I or a pharmaceutically acceptable salt or prodrug or solvate thereof.

[0009] 2. The compound for use according to item 1, which is represented by Formulawherein X is selected from the group consisting of H and a pharmaceutically acceptable cation such as Na+or K+.

[0010] 3. The compound for use according to item 2, wherein the pharmaceutically acceptable cation is Na+.

[0011] 4. The compound for use according to any one of items 1 to 3, wherein the bacterial infection is associated with Staphylococcus aureus, preferably methicillin-resistant Staphylococcus aureus.

[0012] 5. The compound for use according to any one of items 1 to 4, wherein the compound is administered at a dosage of 40 mg / day to 3000mg / day.

[0013] 6. The compound for use according to any one of items 1 to 5, wherein the compound is administered orally or intravenously.

[0014] 7. The compound for use according to any one of claims 1 to 6, wherein the patient is a human.

[0015] 8. The compound for use according to any one of items 1 to 7, wherein the method comprises administration of one or more further antibiotics or antibacterial agents.

[0016] 9. The compound for use according to any one of items 1 to 8, wherein the bacterial infection is selected from the group consisting of acute bacterial skin and skin structure infections, bone and joint infections and diabetic foot infections.

[0017] 10. A method of treatinga bacterial infection selected from infections associated with a Staphylococcal species, comprising administering to a patient in need thereof a pharmaceutically effective amount of a compound represented by Formula I:or a pharmaceutically acceptable salt or prodrug or solvate thereof, wherein preferably the method is specified in any one of items 2 to 9.

[0018] 11. A pharmaceutical composition for use in a method of treating a bacterial infection selected from infections associated with a Staphylococcal species in a patient, wherein the composition comprises the compound specified in any one of items 1 to 9 and a pharmaceutically acceptable excipient.

[0019] 12. A compound represented by the following Formula II:wherein X is a pharmaceutically acceptable cation such as Na+or K+.

[0020] 13. A compound according to item 12, wherein the pharmaceutically acceptable cation is Na+.

[0021] 14. A pharmaceutical composition comprising the compound specified in item12 or 13 and a pharmaceutically acceptable excipient.Brief Description of the Drawings

[0022] Figure 1 shows plasma concentration of compound 1 (closed symbols, unbroken lines) and compound 2 (open symbols, dashed lines) in the Staphylococcus aureus neutropenic murine thigh model according to Pharmacological Example 2. Doses listed in the figure are compound 1 equivalent values. The letter 'h' stands for 'hour' and the abbreviation 'LLOQ' stands for 'lower limit of quantification'.

[0023] Figure 2 shows in vivo efficacy of compound 1 (dosed as compound 2, data presented in open squares) in a Staphylococcus aureus neutropenic murine thigh infection model according to Pharmacological Example 3. Vehicle (open circes) and linezolid (open triangles) were introduced as a control. The abbreviation 'CFU' stand for 'colony forming unit'.Detailed Description of the InventionDefinitions

[0024] The following definitions are provided to assist the reader. Unless otherwise defined, all terms of art, notations, and other scientific or medical terms or terminology used herein are intended to have the meanings commonly understood by those of skill in thechemical and medical arts. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not be construed as representing a substantial difference over the definition of the term as generally understood in the art.

[0025] In some embodiments, the term "about" refers to a deviation of ± 10 % from the recited value. When the word "about" is used herein in reference to a number, it should be understood that still another embodiment of the invention includes that number not modified by the presence of the word "about".

[0026] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0027] The term "including" is used to mean "including but not limited to". "Including" and "including but not limited to" are used interchangeably. The term "comprising" is used to have the same meaning as "including". The term "consisting of" is used to indicate that the listed element(s) is / are present but no other unmentioned elements. The term "comprising" is used to include the meaning of "consisting of" as a preferred embodiment.

[0028] "Administering" or "administration of" a drug to a patient (and grammatical equivalents of this phrase) refers to direct administration, which may be administration to a patient by a medical professional or may be self-administration, and / or indirect administration, which may be the act of prescribing a drug. E.g., a physician who instructs a patient to self-administer a drug or provides a patient with a prescription for a drug is administering the drug to the patient.

[0029] The terms "treatment" and "therapy", as used in the present application, refer to a set of hygienic, pharmacological, surgical and / or physical means used with the intent to cure and / or alleviate a disease and / or symptoms with the goal of remediating the health problem. The terms "treatment" and "therapy" include preventive and curative methods, since both are directed to the maintenance and / or reestablishment of the health of an individual or animal. Regardless of the origin of the symptoms, disease and disability, the administration of a suitable medicament to prevent, alleviate and / or cure a health problem should be interpreted as a form of treatment or therapy within the context of this application.

[0030] The term "treating" includes any significant effect, e.g., lessening, reducing, modulating, or eliminating, that results in the improvement of the condition, disease, disorder and the like.

[0031] The term "prophylactic" or "therapeutic" treatment is art-recognized and refers to administration to the host of one or more of the subject compositions. If it is administered prior to clinical manifestation of the unwanted condition (e.g., disease or other unwanted state of the host animal) then the treatment is prophylactic, i.e., it protects the host against developing the unwanted condition, whereas if administered after manifestation of the unwanted condition, the treatment is therapeutic (i.e., it is intended to diminish, ameliorate or maintain the existing unwanted condition or side effects therefrom).

[0032] A "patient," "subject" or "host" to be treated by the subject method may mean either a human or non-human animal. Non-human animals include companion animals (e.g. cats, dogs) and animals raised for consumption (i.e. food animals), such as cows, pigs, chickens. Non-human animals are preferably mammals. In one aspect, the patient, subject or host is diagnosed with a bacterial infection and is in need of treatment.

[0033] The term "mammal" is known in the art, and exemplary mammals include humans, primates, bovines, porcines, canines, felines, and rodents (e.g., mice and rats).

[0034] "Dose" and "dosage" refer to a specific amount of active or therapeutic agents for administration. Such amounts are included in a "dosage form," which refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active agent calculated to produce the desired onset, tolerability, and therapeutic effects, in association with one or more suitable pharmaceutical excipients such as carriers.

[0035] "Unit dosage form" as used herein refers to a physically discrete unit of therapeutic formulation appropriate for the subject to be treated. It will be understood, however, that the total daily usage of the compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular subject or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of specific active agent employed; specific composition employed; age, body weight, general health, sex and diet of the subject; time of administration, and rate of excretion of the specific active agent employed; duration of the treatment; drugs and / or additional therapies used in combinationor coincidental with specific compound(s) employed, and like factors well known in the medical arts.

[0036] The term "Fabl" is art-recognized and refers to the bacterial enzyme believed to function as an enoyl-acyl carrier protein (AGP) reductase in the final step of the four reactions involved in each cycle of bacterial fatty acid biosynthesis. This enzyme is believed to be widely distributed in bacteria and plants.

[0037] The term "enzyme inhibitor" refers to any compound that prevents an enzyme from effectively carrying out its respective biochemical roles. Therefore a "Fabl inhibitor" is any compound that inhibits Fabl from carrying out its biochemical role. The amount of inhibition of the enzyme by any such compound will vary and is described herein and elsewhere.

[0038] The terms "antibiotic agent", "antibiotic" or "antibacterial agent" shall mean any drug that is useful in treating, preventing, or otherwise reducing the severity of any bacterial disorder, or any complications thereof, including any of the conditions, disease, or complications arising therefrom and / or described herein. An "antibiotic agent" or "antibiotic" can kill or inhibit the growth of microorganisms, such as bacteria, fungus or parasites. On the other hand, an "antibacterial agent" specifically acts on bacteria only. Antibiotics and antibacterial agents include, for example, cephalosporins, quinolones and fluoroquinolones, penicillins and beta lactamase inhibitors, carbapenems, monobactams, macrolides, lipopeptides and lincosamides, glycopeptides, rifampin, oxazolidinones, tetracyclines, aminoglycosides, streptogramins, sulfonamides, and the like. Other antibiotic or antibacterial agents are disclosed herein, and are known to those of skill in the art. In certain embodiments, the terms "antibiotic" and "antibacterial agent" do not include an agent that is a Fabl inhibitor, so that the combinations of the present invention in certain instances will include one agent that is a Fabl inhibitor and another agent that is not. In a preferable embodiment, the terms "antibiotic" and "antibacterial agent" do not include an agent that is a compound of Formula I, so that the combinations of the present invention in certain instances will include one agent that is a compound of Formula I and another agent that is not.

[0039] The term "drug" as used herein refers to any substance falling within at least one of the definitions given in Article 1, Items 2(a), 2(b) or 3a. of Directive 2001 / 83 / EC of November 6, 2001 in the version of November 16, 2012 or in Article 1, Items 2(a) or 2(b) ofDirective 2001 / 82 / EC of November 6, 2001 in the version of August 7, 2009 and in Article 2 of Regulation (EC) No. 726 / 2004 of March 31, 2004.

[0040] The term "chiral" is art-recognized and refers to molecules which have the property of non-superimposability of the mirror image partner, while the term "achiral" refers to molecules which are superimposable on their mirror image partner. A "prochiral molecule" is a molecule which has the potential to be converted to a chiral molecule in a particular process.

[0041] The compounds of the disclosure may contain one or more chiral centers and / or double bonds and, therefore, exist as geometric isomers, enantiomers or diastereomers. The enantiomer and diastereomers may be designated by the symbols "(+)", "(-)", "R" or "S," depending on the configuration of substituents around the stereogenic carbon atom, but the skilled artisan will recognize that a structure may denote one or more chiral centers implicitly. Mixtures of enantiomers or diastereomers may be designated "(±)" in nomenclature, but the skilled artisan will recognize that a structure may denote a chiral center implicitly. Geometric isomers, resulting from the arrangement of substituents around a carbon-carbon double bond or arrangement of substituents around a cycloalkyl or heterocyclic ring, can also exist in the compounds of the present invention.

[0042] Substituents around a carbon-carbon double bond are designated as being in the "Z" or "E" configuration wherein the terms "Z" and "E" are used in accordance with IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both the "£" and “Z" isomers. Substituents around a carbon-carbon double bond alternatively can be referred to as "cis" or "trans," where "cis" represents substituents on the same side of the double bond and "trans" represents substituents on opposite sides of the double bond. The arrangement of substituents around a carbocyclic ring can also be designated as "cis" or "trans." The term "cis" represents substituents on the same side of the plane of the ring and the term "trans" represents substituents on opposite sides of the plane of the ring. Mixtures of compounds wherein the substituents are disposed on both the same and opposite sides of plane of the ring are designated "cis / trans" or "Z / E."

[0043] The term "stereoisomers" when used herein consist of all geometric isomers, enantiomers or diastereomers. The present invention encompasses various stereoisomers of these compounds and mixtures thereof. Conformational isomers and rotamers of disclosed compounds are also contemplated.

[0044] It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction.

[0045] The term "substituted" is also contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein above, e.g. in connection with substituted alkyls. The permissible substituents may be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. In this context, the term "permissible substituents" means any substituent that can be bonded to the core molecule without contravening general principles of chemical bond formation such as the maximum number of valence electrons for an atom of interest, and without making the compound so toxic for the patient that inacceptable toxicity is found even at the minimum dosage required for achieving a therapeutic effect.

[0046] For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.

[0047] Unless specified otherwise, the term "protective group" is used herein to characterize a group that is bonded to a functional group to prevent this functional group from participating in a contemplated chemical reaction. The protective group must be inert under the conditions of the contemplated chemical reaction, but it must be possible to remove the protetive group from the compound such that no further transformations take place in other parts of the molecule. Suitable protective groups are described for each functional group in "Greene's Protective Groups in Organic Synthesis", Peter G. M. Wuts, Theodora W. Greene, John Wiley & Sons, 20 Dec 2012.

[0048] The term "prodrug" refers to a derivative of an active compound (drug) that undergoes a transformation under the conditions of use, such as within the body, to releasethe active drug. Prodrugs are frequently, but not necessarily, pharmacologically inactive until converted into the active drug. For example, a prodrug of the compound of Formula I may be a compound wherein the hydroxyl group in the Formula I is converted into a phosphate group. In a preferable embodiment, a prodrug of the compound of Formula I is represented by the Formula II.

[0049] The term "pharmaceutically-acceptable salts" is art-recognized and refers to the relatively non-toxic, inorganic and organic acid addition salts, or inorganic or organic base addition salts of compounds, including, for example, those contained in compositions of the present invention, and including those present in other approved drugs (wherein approval may be by any competent authority in the EU, USA, CA, JP, CN or KR). Pharmaceutically acceptable salts are meant to be encompassed by the present invention. Hence, all references to compounds of the invention are to be understood as references not only to the compounds as such, but also to pharmaceutically acceptable salts of the respective compounds. According to one aspect, the pharmaceutically acceptable salts may be selected from the salts acknowledged as pharmaceutically acceptable in the literature at the filing date, and in particular as described in G.S. Paulekuhn et al. in J. Med. Chem. 2007, 50, 6665-6672 and references cited therein. If the compound of the invention is an acidic compound, and in particular a prodrug containing a phosphate group, said compound of the invention may be provided in the form of pharmaceutically acceptable salts, wherein the compound forms the anion part and the counterion is selected from Na+, K+, Mg2+, Ca2+, NHs+(thereby forming a "pharmaceutically acceptable cation"), or the protonated forms of the organic bases ethanolamine, meglumine, tromethamine (i.e. 2-amino-2-(hydroxymethyl)propane-l,3-diol), and deanol (i.e. 2-(dimethylamino)ethanol). In a preferable embodiment, the compound of the present invention forms the anion part and the counterion is not meglumine. The systematic IUPAC name of meglumine is (2 / ?,3 / ?,4 / ?,5S)-6-(methylamino)hexane-l,2,3,4,5- pentol. The stoichiometry of the salts is not particularly restricted. For instance, salts may be formed with the phosphate prodrugs of the invention (having 2 acidic protons) in any stoichiometric ratio of from 0 to 2 equivalents. Generally, pharmaceutically acceptable salts are formed such that the net charge of the salt is zero, i.e. that the total number of positive charges equals the total number of negative charges. Pharmaceutically acceptable salts can be prepared from compounds of Formula I by conventional methods. For example, the salts may be prepared by treating a compound of Formula I with an aqueous solution of the desiredpharmaceutically acceptable cation and evaporating the resulting solution to dryness. Alternatively, a lower alkyl alcohol solution of the compound of Formula I may be mixed with an alkoxide of the desired metal and subsequently evaporating the solution to dryness.

[0050] The term "solvate" is art-recognized and refers to an aggregate that consists of an active compound (drug) and one or more solvent molecules. For example, a solvate within the meaning of the present invention may refer to an aggregate of a compound of Formula I and one or more solvent molecules. Suitable solvents are for example pharmaceutically acceptable solvents such as water, ethanol, and the like.

[0051] The term "bioavailable" is art-recognized and refers to a form of the subject disclosure that allows for it, or a portion of the amount administered, to be absorbed by, incorporated to, or otherwise physiologically available to a subject or patient to whom it is administered.

[0052] The term "pharmaceutically acceptable carrier" is art-recognized and refers to a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting any subject composition or component thereof from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the subject composition and its components and not injurious to the patient. Some examples of materials which may serve as pharmaceutically acceptable carriers include: (1) sugars, such as dextrose, lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch as well as starch derivatives such as cyclodextrins and modified cyclodextrins including preferably (2-hydroxypropyl)-|3-cyclodextrin and sulfobutylether-|3-cyclodextrin; (3) cellulose, and its derivatives, such as microcrystalline cellulose, sodium carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropylmethyl cellulose (HPMC), and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) matrix-forming polymeric excipients such as polyvinyl pyrrolidine (PVP), e.g. PVP K30, acrylic polymers and copolymers such as the different grades of Eudragit and preferably Eurdragit L100, hydroxypropylmethyl cellulose acetate succinate (HPMCAS), other copolymers such as polyethylene glycol-based copolymers like Soluplus; (9) excipients, such as cocoa butter and suppository waxes; (10) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (11) glycols, such as propylene glycol; (12) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (13) esters, such as ethyl oleate, glycerylbehenate and ethyl laurate; (14) agar; (15) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (16) alginic acid; (17) pyrogen-free water; (18) isotonic saline; (19) Ringer's solution; (20) ethyl alcohol; (21) phosphate buffer solutions; and (22) other non-toxic compatible substances employed in pharmaceutical formulations. The disclosed excipients may serve more than one function. For example, fillers or binders may also be disintegrants, glidants, anti-adherents, lubricants, sweeteners and the like.

[0053] The term "solvent" is used herein to mean a liquid chemical substance that is capable of dissolving a significant quantity of another substance of interest, the "solute", to thereby generate a clear homogeneous solution. The term "significant quantity" is determined by the intended use of the solution in such a manner that the intended use must be possible by the dissolved quantity of the solute. For instance, if it is intended to administer a compound of the present invention in the form of a solution by injection, the solvent must be capable of dissolving the compound in such amounts, to make administration of a therapeutic dose possible.

[0054] The terms "acid" and "base" are used to have their conventional meanings as proton donators and proton acceptors, respectively (i.e. Broensted acids and bases). A "strong base" is meant to be any base having a basicity of t-BuOK in THF or stronger. A "mild acid" is meant to be any acid having acidity of IM H2SO4 or weaker.

[0055] The term "therapeutic effect" is art-recognized and refers to a local or systemic effect in animals, particularly mammals, and more particularly humans caused by a pharmacologically active substance. The term thus means any measurable effect in the diagnosis, cure, mitigation, treatment or prevention of disease or in the enhancement of desirable physical or mental development and / or conditions in an animal or human. The phrase "therapeutically-effective amount" means that amount of such a substance that produces some desired local or systemic effect at a reasonable benefit / risk ratio applicable to any treatment. The therapeutically effective amount of such substance will vary depending upon the subject and disease condition being treated, the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. For example, certain compositions of the present invention may be administered in a sufficient amount to produce a therapeutic effect at a reasonable benefit / risk ratio applicable to such treatment.

[0056] The term "ICso" is art-recognised and refers to the effectiveness of a substance in inhibiting a given biological or biochemical process (or component of a process, i.e. an enzyme, cell, cell receptor or microorganism). IC50 represents the concentration of a drug e.g. a compound of the invention, that is required for 50% inhibition in vitro.

[0057] The term "MIC" is art-recognised and refers to the Minimum Inhibitory Concentration, that is the lowest concentration of an antimicrobial that will inhibit the visible growth of a microorganism following overnight incubation, usually reported as mg / L or pg / mL.

[0058] The term "ED50" is art-recognized. In certain embodiments, ED50 means the effective dose of a drug which produces 50% of its maximum response or effect, or alternatively, the dose which produces a pre-determined response in 50% of test subjects or preparations.

[0059] The term "LD50" is art-recognized. In certain embodiments, LD50 means the dose of a drug which is lethal in 50% of test subjects.

[0060] The term "therapeutic index" is an art-recognized term that refers to the therapeutic index of a drug, defined as ED50 / LD50.

[0061] The term "Ki" is art-recognized and refers to the dissociation constant of the enzyme-inhibitor complex.

[0062] The term "antimicrobial" is art-recognized and refers to the ability of the compounds disclosed herein to prevent, inhibit or destroy the growth of microbes such as bacteria, fungi, protozoa and viruses.

[0063] The term "antibacterial" is art-recognized and refers to the ability of the compounds disclosed herein to prevent, inhibit or destroy the growth of microbes of bacteria.

[0064] The term "microbe" is art-recognized and refers to a microscopic organism. In certain embodiments the term microbe is applied to bacteria. In other embodiments the term refers to pathogenic forms of a microscopic organism.

[0065] The term "illness" as used herein refers to any illness caused by or related to infection by an organism.

[0066] The term "bacterial illness" as used herein refers to any illness caused by or related to infection by bacteria.

[0067] The term "Staphylococcal species" refers to bacteria species of the Staphylococcus genus belonging to the family of Staphylococcaceae. Examples ofStaphylococcal species are 5. argenteus, S. aureus, S. capitis, S. caprae, S. cohnii, S. hominis, S. schweitzeri, S. simiae, S. auricularis, S. carnosus, S. epidermidis, S. haemolyticus, S. hyicus, S. intermedius, S. lugdunensis, S. saprophyticus, S. sciuri, S. simulans and 5. warneri .

[0068] The term "infections associated with a Staphylococcal species" is used to characterize any infection wherein a Staphylococcal species is present at the site of infection, and it includes infections that are directly or indirectly related with a Staphylococcal species, such as primary and secondary infections with a Staphylococcal species including coinfections and opportunistic infections. Preferably, infections associated with a Staphylococcal species mean infections by a Staphylococcal species. Similarly, the term "infections associated with Staphylococcus aureus" includes infections that are directly or indirectly related with Staphylococcus aureus. Preferably, infections associated with Staphylococcus aureus mean infections by Staphylococcus aureus, more preferably infections by methicillin-resistant Staphylococcus aureus.

[0069] Unless specified otherwise, all reactions described herein are carried out at reaction temperatures that yield the desired target compound and that provide a reasonable compromise between reaction rate and selectivity.Overview

[0070] Surprisingly, it has been found that antibacterial activity against Staphylococcal species, and in particular against Staphylococcus aureus, can be achieved with a compound of Formula I or a prodrug of Formula II. Furthermore, it has been found that the compound of the present invention and its pharmaceutically acceptable salts, prodrugs or solvates have a low MIC with respect to Staphylococcal species while simultaneously being less prone to interact with PDE3 and DAT receptors than other, structurally closely related Fabl inhibitor compounds. This indicates that the compounds of the present invention are not only effective against these types of bacteria, but are also effective at low dosages, thereby minimizing side effects while being very well tolerated.Compounds of the Invention

[0071] The compound of the present invention that is useful in a method of treating a bacterial infection associated with a Staphylococcal species in a patient is represented by the following Formula I, which includes pharmaceutically acceptable salts, prodrugs and solvates thereof:Formula I

[0072] In a preferable embodiment, the compound of the present invention that is useful in a method of treating a bacterial infection associated with a Staphylococcal species is a prodrug represented by the following Formula II:wherein X is selected from a group consisting of H and a pharmaceutically acceptable cation.

[0073] Pharmaceutically acceptable cations include Na+, K+, Mg2+and Ca2+. In a preferable embodiment, the pharmaceutically acceptable cation is Na+or K+. In the most preferable embodiment, the pharmaceutically acceptable cation is Na+.

[0074] The present disclosure contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (d)-isomers, (l)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. However, the carbon-carbon double bond between the pyridine ring and the amide group in the center of the molecule must be in trans configuration, as shown in the above Formula I.

[0075] If, for instance, a particular enantiomer of a compound disclosed herein is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.

[0076] Moreover, individual enantiomers and diastereomers of compound of the present invention can be prepared synthetically from commercially available startingmaterials that contain asymmetric or stereogenic centers, or by preparation of racemic mixtures followed by resolution methods well known to those of ordinary skill in the art. These methods of resolution are exemplified by (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography and liberation of the optically pure product from the auxiliary, (2) salt formation employing an optically active resolving agent, (3) direct separation of the mixture of optical enantiomers on chiral liquid chromatographic columns or (4) kinetic resolution using stereoselective chemical or enzymatic reagents. Racemic mixtures can also be resolved into their component enantiomers by well known methods, such as chiral-phase gas chromatography or crystallizing the compound in a chiral solvent. Stereoselective syntheses, a chemical or enzymatic reaction in which a single reactant forms an unequal mixture of stereoisomers during the creation of a new stereocenter or during the transformation of a pre-existing one, are well known in the art. Stereoselective syntheses encompass both enantio- and diastereoselective transformations. For examples, see Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH: Weinheim, 2009.

[0077] The compounds disclosed here may exist in single or multiple crystalline forms or polymorphs. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in a crystalline form.

[0078] The invention also embraces an isotopically labeled compound of the invention which is as recited herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as2H,3H,13C,14C,15N,180,170,31P,32P,35S,18F, and36CI, respectively. For example, a compound of the invention may have one or more H atom replaced with deuterium.

[0079] Certain isotopically-labeled disclosed compounds (e.g., those labeled with 3H and 14C) are useful in compound and / or substrate tissue distribution assays. Tritiated (i.e., 3H) and carbon-14 (i.e.,14C) isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e.,2H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g.,increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances. Isotopically labeled compounds of the invention can generally be prepared by following procedures analogous to those disclosed in the e.g., Examples herein by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.ProdrugsProdrugs of the compound of the present invention contain at least one prodrug moiety, i.e. a moiety that is cleaved under physiologic conditions to thereby release the active species. Such prodrug moieties may be attached to the compound of Formula I at the hydroxy position or at any one of the secondary amine positions of the 7-membered heterocyclic ring. Suitable prodrug moieties are phospates or other solubilizing moieties as described in "Prodrugs: design and clinical applications" (Rautio et al. Nature Reviews Drug Discovery, 2008, 7, 255). One specific prodrug modification is the conversion of an amine group to an N-oxide group. The above Formula I illustrates suitable positions for prodrug modifications by means of the phosphate moiety. It should be understood that these prodrug moieties may be replaced by other prodrug moieties. According to a preferred embodiment of the present invention, the prodrugs of the invention contain exactly one prodrug moiety. In the most preferred embodiment, the prodrug is represented by Formula II as described above.Pharmaceutically acceptable Salts, Solvates, Polymorphs

[0080] The compound of the present invention may be used in the free form or, alternatively, in the form of pharmaceutically acceptable salts. Acid addition salts are particularly suitable. Particularly effective salts may be hydrochloride salts e.g. hydrochloride or dihydrochloride salts, or fluoroacetate salts e.g. trifluoroacetate salts.

[0081] Furthermore, pharmaceutically acceptable salts of the compound of the present invention include compounds, wherein the compound represented by Formula I or its prodrug forms the anion part and the counterion is selected from Na+, K+, Mg2+, Ca2+(thereby forming a pharmaceutically acceptable cation), or the protonated forms of the organic bases ethanolamine, meglumine, tromethamine (i.e. 2-amino-2- (hydroxymethyl)propane-l,3-diol), and deanol (i.e. 2-(dimethylamino)ethanol). In a preferable embodiment, the compound represented by Formula II forms the anion part and the counterion is not meglumine. The stoichiometry of the salts is not particularly restricted. For instance, salts may be formed with the phosphate prodrugs of the invention (having 2 acidic protons) in any stoichiometric ratio of from 0 to 2 equivalents. Generally,pharmaceutically acceptable salts are formed such that the net charge of the salt is zero, i.e. that the total number of positive charges equals the total number of negative charges.

[0082] The prodrugs of the present invention may also be provided in the free form or in the form of pharmaceutically acceptable salts. Suitable are pharmaceutically acceptable salts well-known to the skilled person, e.g. as described in the literature cited above.

[0083] The compound of the present invention can exist in solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms.

[0084] The compound of the invention may exist in single or multiple crystalline forms or polymorphs. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In another embodiment, the compound is in a crystalline form. Methods of Manufacture

[0085] The compounds of the present invention can be prepared using established organic chemistry synthetic methods and procedures and / or information described in WO 2020 / 099341 Al, the contents of which is hereby incorporated by reference. In particular, the compounds of the present invention can be prepared following the methods described in the chapter of WO 2020 / 099341 Al named "Methods of manufacture", including the sections "Final Step Amide Coupling", "Manufacture of Right-Hand Side Precursor", "Manufacture of H-NA1A2 Precursor" and "Final Step Heck Coupling".

[0086] The preparation of prodrugs of the compounds of the invention is typically accomplished by converting the compound of Formula I to a compound of the same structure except that the hydrogen atom of the hydroxy moiety or any one of the secondary amine moieties of the 7-membered heterocyclic ring represents a prodrug moiety that is cleavable under physiologic conditions for instance a phosphate-containing group as specified above. The prodrug moiety is preferably a phosphate moiety. Such prodrug moieties and suitable reaction conditions for manufacturing phosphate prodrugs are described in WO 2020 / 099341 Al. When seeking to include a prodrug moiety to only one of the hydroxy moiety or any one of the secondary amine moieties of the 7-membered heterocyclic ring, the other two reactive groups would have to be suitably protected.Pharmaceutical Compositions

[0087] The compounds of the present invention may be comprised in pharmaceutical compositions together with a pharmaceutically acceptable excipient.

[0088] Pharmaceutical compositions of the disclosure may be administered by various means, depending on their intended use, as is well known in the art. For example, if compositions of the disclosure are to be administered orally, they may be formulated as tablets, capsules, granules, powders or syrups. Alternatively, formulations disclosed herein may be administered parenterally as injections (intravenous, intramuscular or subcutaneous), drop infusion preparations or suppositories. For application by the ophthalmic mucous membrane route, the compositions disclosed herein may be formulated as eye drops or eye ointments. These formulations may be prepared by conventional means, and, if desired, the compositions may be mixed with any conventional additive, such as an excipient, a binder, a disintegrating agent, a lubricant, a corrigent, a solubilizing agent, a suspension aid, an emulsifying agent or a coating agent. The disclosed excipients may serve more than one function. For example, fillers or binders may also be disintegrants, glidants, anti-adherents, lubricants, sweeteners and the like.

[0089] In formulations of the disclosure, wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may be present in the formulated agents.

[0090] Subject compositions may be suitable for oral, nasal (e.g., by inhalation using a dry powder formulation or a nebulized formulation), topical (including buccal and sublingual), pulmonary (including aerosol administration), rectal, vaginal, aerosol and / or parenteral (e.g., by injection, for example, intravenous, intramuscular, or subcutaneous injection) administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of a compound disclosed herein that may be combined with a carrier material to produce a single dose may vary depending upon the identity of the compound, the subject being treated, and the particular mode of administration.

[0091] Methods of preparing these formulations include the step of bringing into association compositions of the disclosure with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimatelybringing into association agents with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0092] Formulations suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or nonaqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia), each containing a predetermined amount of a subject composition thereof as an active ingredient. Compositions of the disclosure may also be administered as a bolus, electuary, or paste.

[0093] In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the subject composition is mixed with one or more pharmaceutically acceptable excipients selected from: (1) fillers or extenders, such as starches, dextrose, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, celluloses (e.g., microcrystalline cellulose, methyl cellulose, hydroxypropylmethyl cellulose (HPMC) and carboxymethylcellulose), alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as croscarmellose sodium, sodium carboxymethyl starch (sodium starch glycolate), crosslinked polyvinylpurrolidone (crospovidone), gellan gum, xanthan gum, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid and sodium alginate, certain silicates and especially calcium silicate, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) coloring agents; (11) complexing agents such as cyclodextrins and modified cyclodextrins including preferably (2-hydroxypropyl)-|3-cyclodextrin and sulfobutylether-p-cyclodextrin; (12) matrix-forming polymeric excipients such as polyvinyl pyrrolidine (PVP), e.g. PVP K30, acrylic polymers and co-polymers such as the different grades of Eudragit and preferably Eudragit L100, hydroxypropylmethyl cellulose acetate succinate (HPMCAS), other copolymers such as polyethylene glycol-based copolymers like Soluplus; and (13) carriers, such as sodium citrate or dicalcium phosphate. In the case ofcapsules, tablets and pills, the compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like. The disclosed excipients may serve more than one function. For example, fillers or binders may also be disintegrants, glidants, anti-adherents, lubricants, sweeteners and the like. It is possible in accordance with the present invention to use two or more excipients, wherein said two or more excipients may belong to the same and / or different categories. There is no restriction in this respect.Preferred Oral FormulationsBinders

[0094] A binder is advantageously used for increasing the particle size of active ingredient alone or with excipients and improve its handling properties. There is no particular limitation on the binder material that can be employed in the present invention.

[0095] Suitable binder materials include povidone (polyvinylpyrrolidone), copovidone (Poly(l-vinylpyrrolidone-co-vinyl acetate)), maltodextrin, poloxamer (a block copolymer with a first polyethylene oxide) block, a second and central polypropylene oxide) block and a third polyethylene oxide) block), polyethylene glycol, polyethylene oxide, magnesium aluminosilicate, gelatin, acacia, alginic acid, carbomer (e.g. carbopol), dextrin, dextrates (a purified mixture of saccharides developed from the controlled enzymatic hydrolysis of starch), guar gum, hydrogenated vegetable oil, liquid glucose, wax, starch (pregelatinized and plain), sodium alginate and mixtures thereof.

[0096] The use of povidone and copovidone is preferred.

[0097] The binder may be present in a relative amount of from 0.5 wt% to 15 wt%, preferably from 1 wt% to 12 wt% and more preferably 4 wt% to 10 wt%.Diluents

[0098] Diluents are advantageously used for increasing the bulk of the pharmaceutical composition and for facilitating handling of the composition. There is no particular limitation on the diluent material that can be employed in the present invention.

[0099] Suitable diluent materials include mannitol, isomalt, histidine, lactose (including anhydrous or monohydrate forms), calcium phosphate (including dibasic and tribasic calcium phosphate), calcium carbonate, calcium sulfate, sucrose, fructose, maltose, xylitol, sorbitol, maltitol, aluminium silicate, dextrose, starch (pregelatinized or plain),glucose, dextrates (a purified mixture of saccharides developed from the controlled enzymatic hydrolysis of starch), magnesium carbonate, and mixtures thereof.

[0100] The use of mannitol, xylitol, sorbitol, isomalt and / or histidine is preferred. Mannitol is particularly preferred.

[0101] The diluent may be present in a relative amount that is not particularly restricted. Suitable amounts may range from 2 wt% to 85 wt%, preferably from 8 wt% to 80 wt% and more preferably 10 wt% to 50 wt%.Surfactant

[0102] A surfactant may advantageously be used for assisting wettability of the tablet and of the active ingredient. The surfactant is an optional but preferred component. There is no particular limitation on the surfactant material that can be employed in the present invention.

[0103] Suitable surfactant materials include sodium lauryl sulfate, poloxamer, sodium docusate, sorbitan esters, polyethylene oxide, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80 (ethoxylated sorbitan esterified with fatty acids wherein the number indicates the number of repeating units of polyethylene glycol), and mixtures thereof.

[0104] The use of sodium lauryl sulfate is preferred.

[0105] The surfactant may be present in a relative amount that is not particularly restricted. Suitable amounts may range from 0 wt% or more to 7 wt%, preferably from 0.1 wt% to 6.5 wt% and more preferably 1 wt% to 6 wt%.Disintegrant

[0106] A disintegrant is used for accelerating disintegration of the pharmaceutical composition to thereby assist in dissolution and uptake of the active ingredient. There is no particular limitation on the disintegrant material that can be employed in the present invention.

[0107] Suitable disintegrant materials include crosslinked polyvinylpurrolidone (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), croscarmellose sodium, gellan gum, xanthan gum, magnesium aluminosilicate, sodium alginate, pregelatinized starch, alginic acid, guar gum, homo- and copolymers of (meth)acrylic acid and salts thereof such as polacri II i n potassium, and mixtures thereof.

[0108] The use of crospovidone is preferred.

[0109] The disintegrant may be present in a relative amount that is not particularly restricted. Suitable amounts may range from 0 wt% or more to 20 wt%, preferably from 1 wt% to 15 wt% and more preferably 2 wt% to 10 wt%.Glidant

[0110] A glidant is advantageously used for improving flowability of the pharmaceutical composition to thereby improve its handling properties. The glidant is an optional but preferred component. There is no particular limitation on the glidant material that can be employed in the present invention.

[0111] Suitable glidant materials include colloidal silica dioxide, magnesium oxide, magnesium silicate, tribasic calcium phosphate, and mixtures thereof.

[0112] The use of colloidal silica dioxide is preferred.

[0113] The glidant may be present in a relative amount that is not particularly restricted. Suitable amounts may range from 0 wt% or more to 5 wt%, preferably from 0.1 wt% to 4 wt% and more preferably 0.2 wt% to 1 wt%.Lubricant

[0114] Lubricants are advantageously used to facilitate tableting, in particular by preventing sticking of the tablets to the tablet punch. The lubricant is an optional but preferred component. There is no particular limitation on the lubricant material that can be employed in the present invention.

[0115] Suitable lubricant materials include magnesium stearate, sodium stearyl fumarate, talc, stearic acid, leucine, poloxamer, polyethylene glycol, glyceryl behenate, glycerin monostearate, magnesium lauryl sulfate, sucrose esters of fatty acids, calcium stearate, aluminum stearate, hydrogenated castor oil, hydrogenated vegetable oil, mineral oil, sodium benzoate, zinc stearate, palmitic acid, carnauba wax, sodium lauryl sulfate, polyoxyethylene monostearates, calcium silicate, and mixtures thereof.

[0116] The use of a lubricant selected from magnesium stearate and sodium stearyl fumarate, and combinations thereof is preferred.

[0117] The lubricant may be present in a relative amount that is not particularly restricted. Suitable amounts may range from 0 wt% or more to 7 wt%, preferably from 0.1 wt% to 4 wt% and more preferably 0.5 wt% to 3.5 wt%.Matrix forming polymers and copolymers

[0118] Suitable matrix-forming polymers and copolymers include polyvinyl pyrrolidine (PVP), acrylic polymers and co-polymers such as the different grades of Eudragit, hydroxypropylmethyl cellulose acetate succinate (HPMCAS), as well as other copolymers such as polyethylene glycol-based copolymers like Soluplus.

[0119] Preferred matrix-forming polymers and copolymers are HPMC AS and Soluplus.

[0120] The matrix-forming polymers and copolymers may be present in a relative amount that is not particularly restricted. Suitable amounts may range from 0.1 g to 10 g, preferably from 0.2 g to 5 g and more preferably from 0.3 g to 4 g.Complexing agents

[0121] Suitable complexing agents include cyclodextrins and modified cyclodextrins.

[0122] Preferred complexing agents include (2-hydroxypropyl)-|3-cyclodextrin and sulfobutylether-p-cyclodextrin.

[0123] The complexing agents may be present in a relative amount that is not particularly restricted. Suitable amounts may range from 0.1 g to 24 g, preferably from 0.1 g to 10 g and more preferably from 0.1 g to 5 g.Other types of excipients

[0124] The composition of the present invention may contain further excipients that are commonly used in the art.

[0125] Such further excipients may include release rate modifiers, plasticizer, film forming agent, colorant, anti-tacking agent and / or pigment for coating the compositions of the present invention. Further types of excipients, which may be present, include flavoring agents, sweeteners, antioxidants, absorption accelerators and / or bulking agents. Relative amounts of such excipients are not particularly limited. They may be determined by the skilled person based on common general knowledge and routine procedures.

[0126] Film forming agents are advantageously used for providing the tablets of the invention with a coherent coating. Suitable film forming agents include isomalt, polyvinyl alcohol, polyethylene glycol, maltodextrin, sucrose, xylitol, maltitol, enteric coating agents such as materials selected from the group consisting of methyl acrylate-methacrylic acid copolymers, polyvinyl acetate phthalate (PVAP), methyl methacrylate-methacrylic acid copolymers, shellac, sodium alginate and zein. It is preferred to use a combination of film forming agents comprising polyvinyl alcohol and one or more second agents selected fromisomalt, maltodextrin, sucrose, xylitol, and maltitol. It is particularly preferred to use a combination of film forming agents comprising at least polyvinyl alcohol and isomalt.

[0127] Suitable plasticizers include sorbitol, triacetin, poloxamer, polyethylene glycol, glycerin, propylene glycol, polyethylene glycol monomethyl ether, acetyl tributyl citrate, acetyl triethyl citrate, castor oil, glyceryl monostearate, diacetylated monoglyerides, dibutyl sebacate, diethyl phthalate, triethyl citrate, and tributyl citrate.

[0128] For each of the above-mentioned categories of excipients it is possible to use only a single substance or a combination of two or more substances belonging to the same category. Of course, it is not necessary that members of each and every category are present.

[0129] Formulations and compositions of the invention may include the compounds disclosed herein in the form of particles of amorphous substance or in any crystalline form. The particle size is not particularly limited. For instance, formulations and compositions may include micronized crystals of the disclosed compounds. Micronization may be performed on crystals of the compounds alone, or on a mixture of crystals and a part or whole of pharmaceutical excipients or carriers. Mean particle size of micronized crystals of a disclosed compound may be for example about 5 to about 200 microns, or about 10 to about 110 microns. The compounds of the invention may also be present in the form of a molecular dispersion within a polymeric matrix. In yet another embodiment, the compounds of the invention may be complexed with suitable complexing agents such as cyclodextrins.

[0130] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin, microcrystalline cellulose, or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the subject composition moistened with an inert liquid diluent. Tablets, and other solid dosage forms, such as dragees, capsules, pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. The disclosed excipients may serve more than one function. For example, fillers or binders may also be disintegrants, glidants, anti-adherents, lubricants, sweeteners and the like.

[0131] It will be appreciated that a disclosed composition may include lyophilized or freeze dried compounds disclosed herein. For example, disclosed herein are compositions that disclosed compounds crystalline and / or amorphous powder forms. Such forms may be reconstituted for use as e.g., an aqueous composition.

[0132] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the subject composition, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, cyclodextrins and mixtures thereof.

[0133] Suspensions, in addition to the subject composition, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum meta hydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0134] Formulations for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing a subject composition with one or more suitable non-irritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the body cavity and release the active agent. Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0135] Dosage forms for transdermal administration of a subject composition includes powders, sprays, ointments, pastes, creams, lotions, gels, solutions, and patches. The active component may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

[0136] The ointments, pastes, creams and gels may contain, in addition to a subject composition, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch,tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0137] Powders and sprays may contain, in addition to a subject composition, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0138] Compositions and compounds of the disclosure may alternatively be administered by aerosol. This is accomplished by preparing an aqueous aerosol, liposomal preparation or solid particles containing the compound. A non-aqueous (e.g., fluorocarbon propellant) suspension could be used. Sonic nebulizers may be used because they minimize exposing the agent to shear, which may result in degradation of the compounds contained in the subject compositions.

[0139] Ordinarily, an aqueous aerosol is made by formulating an aqueous solution or suspension of a subject composition together with conventional pharmaceutically acceptable carriers and stabilizers. The carriers and stabilizers vary with the requirements of the particular subject composition, but typically include non-ionic surfactants (Tweens, pluronics, or polyethylene glycol), innocuous proteins like serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars or sugar alcohols. Aerosols generally are prepared from isotonic solutions.

[0140] It should be noted that excipients given as examples may have more than one function. For example, fillers or binders can also be disintegrants, glidants, anti-adherents, lubricants, sweeteners and the like.

[0141] Pharmaceutical compositions of this disclosure suitable for parenteral administration comprise a subject composition in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents. For example, provided herein is an aqueous composition that includes a disclosed compound, and may further include for example,dextrose (e.g., about 1 to about 10 weight percent dextrose, or about 5 weight percent dextrose in water (D5W).

[0142] Examples of suitable aqueous and non-aqueous carriers which may be employed in the pharmaceutical compositions of the disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate and cyclodextrins. Proper fluidity may be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0143] It will be appreciated that contemplated formulations, such as oral formulations (e.g. a pill or tablet), may be formulated as controlled release formulation, e.g., an immediate release formulation, a delayed release formulation, or a combination thereof.

[0144] In certain embodiments, the subject compounds may be formulated as a tablet, pill, capsule or other appropriate ingestible formulation (collectively hereinafter "tablet"). In certain embodiments, a therapeutic dose may be provided in 10 tablets or fewer. In another example, a therapeutic dose is provided in 50, 40, 30, 20, 15, 10, 5 or 3 tablets.

[0145] In a certain embodiment, a disclosed compound is formulated for oral administration as a tablet, capsule, or an aqueous solution or suspension. In another embodiment of a tablet form the tablets are formulated such that the resulting amount of antibacterial agent (or antibacterial agents) provided in 20 tablets, if taken together (e.g., over time) once administered, would provide a therapeutically effective dose and / or a dose of at least the median effective dose (EDso), e.g., the dose at which at least 50% of individuals exhibited the quantal effect of inhibition of bacterial cell growth or protection (e.g., a statistically significant reduction in infection). In a further embodiment, tablets may be formulated such that the total amount of antibacterial agent (or antibacterial agents) provided upon administration in 10, 5, 2 or 1 tablets would provide a therapeutically effective dose and / or at least an EDso dose to a patient (human or non-human mammal). In other embodiments, the amount of antibacterial agent (or antibacterial agents) provided, upon administration, in 20, 10, 5 or 2 tablets taken in a 24 hour time period would provide a dosage regimen providing, on average, a mean plasma level of the antibacterial agent(s) of a therapeutically effective dose and / or at least the EDso concentration (the concentration for50% of maximal effect of, e.g., inhibiting bacterial cell growth). In other embodiments less than 100 times, 10 times, or 5 times the ED50 is provided. In other embodiments, a single dose of tablets (1-20 tablets) provides about 40 mg to 3000 mg of compound(s). These specified unit dose amounts apply also for other oral dosage forms.

[0146] Likewise, compounds disclosed herein can be formulated for parenteral administration, as for example, for subcutaneous, intramuscular or intravenous injection, e.g., the antibacterial agent can be provided in a sterile solution or suspension (collectively hereinafter "injectable solution"). The injectable solution may be, in some embodiments, formulated such that the amount of antibacterial agent (or antibacterial agents) provided in, for example, in about 0.1 to about 200cc bolus injection, or a dose administered intravenously, would provide a dose of at least the median effective dose, or less than 100 times the ED50, or less than 10 or 5 times the ED50. The injectable solution may be formulated such that the total amount of antibacterial agent (or antibacterial agents) provided (upon administration) in 100, 50, 25, 10, 5, 2.5, or 1 cc injections would provide a therapeutically effective amount and / or an ED50 dose to a patient, or less than 100 times the ED50, or less than 10 or 5 times the ED50. In other embodiments, the amount of antibacterial agent (or antibacterial agents) provided, upon administration, in a total volume of lOOcc, 50, 25, 5 or 2cc to be injected at least twice in a 24 hour time period would provide a dosage regimen providing, on average, a mean plasma level of the antibacterial agent(s) of a therapeutically effective amount and / or at least the ED50 concentration, or less than 100 times the ED50, or less than 10 or 5 times the ED50. In other embodiments, a single dose injection provides about 40 mg to 3000 mg, or about 100 mg to about 1000 mg of antibacterial agent. In case of i.m. administration, the same amount indications apply, in principle, too. However, the upper end of the unit dose range may be lower depending on the solubility of the drug compound and the maximum tolerable amount for injection.

[0147] The compound of the present invention is preferably administered orally or intravenously.Drug combinations

[0148] The compound of the present invention may also be administered in combination with one or more further antibiotics or antibacterial agents.

[0149] Thus, compositions are also contemplated herein that include one or more of the disclosed compounds with a second component. Second components in suchcompositions of the present disclosure are usually an antibiotic agent other than a disclosed compound. Additional components may also be present, including Fabl inhibitors or other antibiotic agents. The contemplated methods of treatment disclosed herein, in some embodiments, may further comprise administering another agent such as one described below. For example, a method of treating a bacterial infection is provided that comprises administering a disclosed compound and further comprises administering an antibiotic agent or antibacterial agent described below. The compound disclosed herein and the second component may be part of the same dosage form or may be formulated in two separate dosage forms. If they are formulated in two separate dosage forms, the dosage form with the second component may be administered at the same time, before or after the dosage form with the compound disclosed herein.

[0150] Non-limiting examples of antibiotic agents that may be used in the antibacterial compositions of the disclosure include polymyxins, cephalosporins, quinolones and fluoroquinolones, penicillins, penicillins and beta lactamase inhibitors, carbepenems, monobactams, macrolides and lincosamines, glycopeptides, rifampin, oxazolidonones, tetracyclines, aminoglycosides, streptogramins, sulfonamides, and others. Each family comprises many members. In a preferred embodiment, the further antibiotic agent is a polymyxin or aminoglycoside.

[0151] Polymyxins refer to a class of non-ribosomal peptide antibiotics that are naturally produced by Gram-positive bacteria such as Paenibacillus polymyxa and that are effective in treating Gram-negative bacterial infections. A typical representative is colistin methanesulfonate, the sodium salt thereof being commercially available as Colistimethat- Natrium. Another typical representative is SPR206, which is disclosed for instance in P. Brown et al., ACS Infect Dis. 2019 Oct 11; 5(10): 1645-1656; doi: 10.1021 / acsinfecdis.9b00217.

[0152] Cephalosporins can be further categorized by generation. Non-limiting examples of cephalosporins by generation include the following. Examples of cephalosporins: First generation compounds include Cefadroxil, Cefazolin, Cephalexin, Cephalothin, Cephapirin, and Cephradine. Second generation compounds include Cefaclor, Cefamandol, Cefonicid, Cefotetan, Cefoxitin, Cefprozil, Ceftmetazole, Cefuroxime, Cefuroxime axetil, and Loracarbef. Third generation include Cefdinir, Ceftibuten, Cefditoren, Cefetamet, Cefpodoxime, Cefprozil, Cefuroxime (axetil), Cefuroxime (sodium),Cefoperazone, Cefixime, Cefotaxime, Cefpodoxime proxetil, Ceftazidime, Ceftizoxime, and Ceftriaxone. Fourth generation compounds include Cefepime.

[0153] Non-limiting examples of quinolones and fluoroquinolones include Cinoxacin, Ciprofloxacin, Enoxacin, Gatifloxacin, Grepafloxacin, Levofloxacin, Lomefloxacin, Moxifloxacin, Nalidixic acid, Norfloxacin, Ofloxacin, Sparfloxacin, Trovafloxacin, Oxolinic acid, Gemifloxacin, and Perfloxacin.

[0154] Non-limiting examples of penicillins include Amoxicillin, Ampicillin, Bacampicillin, Carbenicillin Indanyl, Mezlocillin, Piperacillin, and Ticarcillin.

[0155] Non-limiting examples of penicillins and beta lactamase inhibitors include Amoxicillin-Clavulanic Acid, Ampicillin-Sulbactam, Benzylpenicillin, Cioxacillin, Dicloxacillin, Methicillin, Oxacillin, Penicillin G (Benzathine, Potassium, Procaine), Penicillin V, Piperacillin+Tazobactam, Tica rci II i n+Clavu la nic Acid, and Nafcillin. Non-limiting examples of carbepenems include Imipenem-Cilastatin and Meropenem.

[0156] A non-limiting example of a monobactam includes Aztreonam. Non-limiting examples of macrolides and lincosamines include Azithromycin, Clarithromycin, Clindamycin, Dirithromycin, Erythromycin, Lincomycin, and Troleandomycin. Non-limiting examples of glycopeptides include Teicoplanin and Vancomycin. Non-limiting examples of rifampins include Rifabutin, Rifampin, and Rifapentine. A non-limiting example of oxazolidonones includes Linezolid. Non-limiting examples of tetracyclines include Demeclocycline, Doxycycline, Methacycline, Minocycline, Oxytetracycline, Tetracycline, and Chlortetracycline.

[0157] Non-limiting examples of aminoglycosides include Amikacin, Arbakacin, Gentamicin, Kanamycin, Sisomicin, Arbekacin, Neomycin, Netilmicin, Streptomycin, Tobramycin, and Paromomycin. A non-limiting example of streptogramins includes Quinopristin+Dalfopristin.

[0158] Non-limiting examples of sulfonamides include Mafenide, Silver Sulfadiazine, Sulfacetamide, Sulfadiazine, Sulfamethoxazole, Sulfasalazine, Sulfisoxazole, Trimethoprim- Sulfamethoxazole, and Sulfamethizole.

[0159] Non-limiting examples of other antibiotic agents include Bacitracin, Chloramphenicol, Colistimethate, Fosfomycin, Isoniazid, Methenamine, Metronidazole, Mupirocin, Nitrofurantoin, Nitrofurazone, Novobiocin, Polymyxin B, Spectinomycin, Tobramycin, Tigecycline, Trimethoprim, Colistin, Cycloserine, Capreomycin, Pyrazinamide, para-Aminosalicyclic acid, and Erythromycin ethylsuccinate + sulfisoxazole.Therapeutic UsesMedical indications

[0160] The compounds of the present invention may be used for treating bacterial infections selected from infections associated with a Staphylococcal species in a patient. They are in particular suitable for use in treating bacterial infection associated with Staphylococcus aureus.

[0161] The compounds of the present invention are particularly useful for treating methicillin-resistant Staphylococcus aureus.Patients

[0162] The compounds of the present invention may be used for treating bacterial infections in patients that are human patients or non-human animals, preferably humans and non-human mammals. In a preferred embodiment, the compounds of the present invention may be used for treating bacterial infections in a human patient (in need of treatment).Dosages

[0163] The dosage of any disclosed compound or composition will vary depending on the symptoms, age and body weight of the patient, the nature and severity of the disorder to be treated or prevented, the route of administration, and the form of the subject composition. Any of the subject formulations may be administered in a single dose or in divided doses. Dosages for the compositions may be readily determined by techniques known to those of skill in the art or as taught herein.In certain embodiments, the dosage of the subject compounds will generally be in the range of about 0.01 ng to about 10 g per kg body weight, specifically in the range of about 1 ng to about 0.1 g per kg, and more specifically in the range of about 100 ng to about 10 mg per kg.

[0164] The compound of the present invention may generally be administered at a dosage of 40 mg / day to 3000 mg / day. In a preferable embodiment, the compound of the present invention is administered at a dosage of 50 mg / day to 3000 mg / day, more preferably 100 mg / day to 2000 mg / day, even more preferably 150 mg / day to 1000 mg / day and most preferably 200 mg / day to 500 mg / day.

[0165] If treatment of the patient by the pharmaceutical compositions of the present invention is by means of oral administration, a single unit dose of the pharmaceutical composition of the present invention is typically administered one, two or three times a day.The daily dosage is determined by the physician in accordance with the above guidance taking severity of the infection, gender, weight, age and general condition of the patient into account. Preferred oral daily dosages range from 40 to 3000 mg, preferably 50 mg to 3000 mg, and more preferably 100 mg to 2000 mg. By consequence, typical unit dosages may range from 50 to 2000 mg, depending on the intended frequency of administration.

[0166] In case of parenteral administration (for instance in i.v. or i.m. administration), the pharmaceutical compositions of the present invention are typically administered two, three or more times a day. Preferred daily dosages are in the range of from 40 to 3000 mg, so that typical unit dosages are from 40 to 3000 mg and preferably 100 to 1000 mg. The upper limits of the specified ranges are subject to their feasibility. For instance, in case of i.m. administration, it may happen that the maximum dose that can be administered in a single shot is restricted due to low solubility and correspondingly increased volume of the drug solution. In such a case, the maximum unit dosages are limited by the maximum tolerated dose.

[0167] An effective dose or amount, and any possible effects on the timing of administration of the formulation, may need to be identified for any particular composition of the disclosure. This may be accomplished by routine experiment as described herein, using one or more groups of animals (preferably at least 5 animals per group), or in human trials if appropriate. The effectiveness of any subject composition and method of treatment or prevention may be assessed by administering the composition and assessing the effect of the administration by measuring one or more applicable indices, and comparing the posttreatment values of these indices to the values of the same indices prior to treatment.

[0168] The precise time of administration and amount of any particular subject composition that will yield the most effective treatment in a given patient will depend upon the activity, pharmacokinetics, and bioavailability of a subject composition, physiological condition of the patient (including age, sex, disease type and stage, general physical condition, responsiveness to a given dosage and type of medication), route of administration, and the like. The guidelines presented herein may be used to optimize the treatment, e.g., determining the optimum time and / or amount of administration, which will require no more than routine experimentation consisting of monitoring the subject and adjusting the dosage and / or timing.

[0169] While the subject is being treated, the health of the patient may be monitored by measuring one or more of the relevant indices at predetermined times during the treatment period. Treatment, including composition, amounts, times of administration and formulation, may be optimized according to the results of such monitoring. The patient may be periodically reevaluated to determine the extent of improvement by measuring the same parameters. Adjustments to the amount(s) of subject composition administered and possibly to the time of administration may be made based on these reevaluations.

[0170] Treatment may be initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage may be increased by small increments until the optimum therapeutic effect is attained.

[0171] The use of the subject compositions may reduce the required dosage for any individual agent contained in the compositions because the onset and duration of effect of the different agents may be complimentary.

[0172] Toxicity and therapeutic efficacy of subject compositions may be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 and the ED50.

[0173] The data obtained from the cell culture assays and animal studies may be used in formulating a range of dosage for use in humans. The dosage of any subject composition lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity and / or that give rise to a statistically significant reduction in infection in at least 50% e.g. 60%, 70%, 80%, 90%, 100%, of individuals with little or no toxicity, wherein little toxicity may for instance mean non serious and / or predictable and transient toxicity. For instance, the dosage of the subject composition may be chosen such that a reasonable benefit / risk ratio is accomplished by the treatment. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For compositions of the disclosure, the therapeutically effective dose may be estimated initially from cell culture assays.Administration frequency

[0174] The compounds and compositions disclosed herein may be administered once or multiple times a day, in particular once daily (qd), twice daily (bid), three times a day (tid) or four times a day (qid).Duration of treatment

[0175] The compounds and compositions disclosed herein may be administered for an unlimited period of time. It is advantageous that they are administered for a period of time to eradicate the bacterial infection completely or at least to such an extent that the patient's immune system can cope with any remaining pathologic bacteria. Typical durations of administration are from 1 day to 2 weeks and especially from 1 to 5 days. In case of i.m. administration, typical durations of administration may range from 1 shot to 4 shots and preferably 1 shot. Multiple shots may be administered on the same day, on consecutive days or on in intervals with days of no administration in between.Abbreviations

[0176] The following abbreviations are used in the present disclosure. g Gram pH Potential of Hydrogen mol Mole mmol Millimole w / w Weight / Weight m / z Mass to charge ratio°C degree CelsiusHPLC High performance liquid chromatography h hour min Minute mL Milliliter eq. EquivalentM Mass pm Mikrometer nm NanometerMe Methyl, CH3MeOH MethanolAcOH Acetic acidTHF TetrahydrofuranEtOH EthanolNH3AmmoniaAq. AqueousRt. Retention timeNMR Nuclear Magnetic Resonance s singlet d doublet t tripletm multiplet dd double of doubletMHz Megahertz ppm parts per millionH ProtonP PhosphorJ Coupling constantMS Mass spectrometryUPLC-MS Ultra-performance liquid chromatography-tandem mass spectrometryLC-MS / MS Liquid Chromatography Tandem Mass SpectrometryDMSO Dimethyl sulfoxideD2O Deuterated waterSCX Strong Cation Exchange ChromatographyLCMS Liquid Chromatography Mass SpectrometryHPLC High-performance liquid chromatographyPDE PhosphodiesteraseDAT Transporter DopamineMIC Minimal Inhibitory ConcentrationIC50 50% Inhibitory concentration pM Mikromolar nM NanomolarKi dissociation constant of the enzyme-inhibitor complexIM or i.m. Intramuscular administrationLLOQ Lower limit of quantificationCFU Colony forming unitsThe following examples are in no way intended to limit the scope of the present invention, but are provided only to illustrate the inventive compounds and their preparation. ExamplesGeneral Procedures

[0177] All starting materials and solvents were obtained either from commercial sources or prepared according to the literature citation. Unless otherwise stated all reactions were stirred. Organic solutions were routinely dried over anhydrous magnesium sulfate or sodium sulfate.

[0178] Column chromatography was performed on pre-packed silica (230-400 mesh, 40-63 pm) cartridges using the eluent indicated. SCX was purchased from Silicycle and treated with IM hydrochloric acid prior to use. Unless stated otherwise the reaction mixture to be purified was first diluted with MeOH and made acidic with a few drops of AcOH. This solution was loaded directly onto the SCX and washed with MeOH. The desired material was then eluted by washing with 0.7 M NH3 in MeOH.Analytical Methods

[0179] Mass Spectrometry Method : Performed based on MS Tune parameters:Routine. ipr as shown below.

[0180] The method is carried out based on the information "MS Method: pos_neg_3mn_30v". Points per peak: 3.846. Total run time: 3.0 min. MS scan, Time 0.00 to 3.00, Mass 100.00 to 900.00 ES+.

[0181] UPLC-MS mass spectra were recorded on a SQ detector Acquity Waters (Waters Corporation, Waters Milford, 34 Maple St., Milford, USA) equipped with software Empower 2 pro in a positive ionization mode. Ionization conditions: Capilary 2.25 KV, Cone 160 V, Source 100 °C, Desolvation 150 °C; Gas flowDesolvation 500 L / h, Cone 50 L / h.XH NMR Spectroscopy

[0182] TH NMR Spectra were acquired on a Bruker Avance III spectrometer at 400 MHz using residual undeuterated solvent as reference.31P NMR Spectroscopy

[0183] 31P NMR Spectra were acquired on Bruker Avance III spectrometer at 161 MHz using phosphoric acid as reference.

[0184] Example 1. Synthesis of (E)-3-((2R,3S)-3-hydroxy-2-methyl-4-oxo-2,3,4,5- tetrahydro-lH-pyrido[2,3-b][l,4]diazepin-8-yl)-N-methyl-N-((3-methylbenzofuran-2- yl)methyl)acrylamide (compound 1)

[0185] The compound 1 of this example was prepared following the procedure as described in Example 25 of WO 2020 / 099341 A.Compound 1

[0186] Example 2. Synthesis of sodium (2R,3S)-2-methyl-8-((E)-3-(methyl((3- methylbenzofuran-2-yl)methyl)amino)-3-oxoprop-l-en-l-yl)-4-oxo-2,3,4,5-tetrahydro-lH- pyrido[2,3-b][l,4]diazepin-3-yl hydrogen phosphate (compound 2)

[0187] To a suspension of compound 1 (1.01 g, 2.40 mmol, 1 eq.) in dry dichloromethane (24.4 mL) at 0 °C was added bis(2-cyanoethyl)-diisopropylphosphoramidite (1.88 mL, 7.21 mmol) and lH-tetrazole (16.0 mL, 7.21 mmol, 0.45M in acetonitrile). The mixture was stirred at 0 °C for 3 h. Then a solution of 0.2 M iodine solution in water / pyridine / tetrahydrofuran (1 / 19 / 80) was added at 0 °C until persistence of iodine coloration. The mixture was stirred at 0 °C for 5 min. A solution of sodium thiosulfate (20% w / w in water) was added until removal of iodide coloration. Water was added and the organic phase was extracted with ethyl acetate. After drying and concentration of the organic phase, the residue was purified on silica gel eluting with 0-10% methanol in dichloromethane and evaporated resulting in bis(2-cyanoethyl) [(2R,3S)-2-methyl-8-[(E)-3- [methyl-[(3-methylbenzofuran-2-yl)methyl]amino]-3-oxo-prop-l-enyl]-4-oxo-l,2,3,5- tetrahydropyrido[2,3-b][l,4]diazepin-3-yl] phosphate (1.35 g, 2.23 mmol, 92.6%) as a yellow solid.

[0188] To a solution of this solid (3.45 g, 5.69 mmol, 1 eq.) in dry acetonitrile (42 mL) at 0 °C were added trimethylsilyl (lE)-2,2,2-trifluoro-N-trimethylsilyl-ethanimidate (5.91 mL, 28.4 mmol) and 2-tert-butyl-l,l,3,3-tetramethylguanidine (7.28 mL, 28.4 mmol). After 10minutes, formic acid (2.15 mL, 56.9 mmol) was added and the mixture was evaporated. Purification by reverse phase C18 column yielded a yellow powder (2.62 g) after lyophilisation, which was dissolved in water (10 mL) and triethylamine (0.29 ml, 2.15 mmol). The solution was stirred at room temperature for two minutes. Sodium 2-ethylhexanoate (2.49 g, 14.6 mmol) was added and the reaction mixture was stirred at for two minutes. The reaction mixture was diluted with acetonitrile (100 mL). The precipitate was filtered and dissolved again in water three times, then finally dissolved in water and lyophilized to afford compound 2 (1.72 g, 3.16 mmol, 56% for 2 steps) as a disodium salt. Rt 1.22 min m / z 501.3 [M + H]+(ES+).XH NMR (400 MHz, D2O): 6 7.94 - 7.86 (m, 1H), 7.47 - 6.77 (m, 7H), 4.71 - 4.53 (m, 2H), 4.23 - 4.12 (m, 1H), 3.05 (s, 1.7H), 2.97 (s, 1.3H), 2.16 - 2.09 (m, 3H), 1.31 - 1.21 (m, 3H).31P (161 MHz, D2O) 6 3.4.Compound 2

[0189] Reference Example 3. Synthesis of (R,E)-3-(3-hydroxy-3-methyl-4-oxo-2,3,4,5- tetrahydro-lH-pyrido[2,3-b][l,4]diazepin-8-yl)-N-methyl-N-((3-methylbenzofuran-2- yl)methyl)acrylamide (compound 3)

[0190] The compound 3 of this reference example was prepared following the procedure as described in Example 8 of WO 2020 / 099341 A.Compound 3

[0191] Reference Example 4. Synthesis of (S,E)-3-(3-hydroxy-4-oxo-2,3,4,5- tetrahydro-lH-pyrido[2,3-b][l,4]diazepin-8-yl)-N-methyl-N-((3-methylbenzofuran-2- yl)methyl)acrylamide (compound 4)

[0192] The compound 4 of this reference example was prepared following the procedure as described in Example 46 of WO 2020 / 099341 A.Compound 4

[0193] Reference Example 5. Synthesis of (S,E)-3-(7-amino-8-oxo-6,7,8,9-tetrahydro-5H-pyrido[2,3-b]azepin-3-yl)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)acrylamide hydrochloride (compound 5)

[0194] The compound 5 of this reference example was prepared following the procedure as described in Example 7 of WO 2021 / 123372 A.Compound 5Pharmacological Examples

[0195] Pharmacological Example 1: Antibacterial, phosphodiesterase PDE3A and transporter dopamine activities of example compounds a) Minimal Inhibitory Concentration (MIC):

[0196] MIC values were determined using broth microdilution according to Clinical and Laboratory Standards Institute (CLSI) guidelines (M07-A11). The reference to the method is the following: CLSI. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard— Eleventh Edition. CLSI document M07- All. CLSI, 950 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087, USA, 2018. b) Phosphodiesterase PDE3A (Human) assay:

[0197] Human recombinant PDE3A expressed in insect Sf9 cells are used. Test compound and / or vehicle is preincubated with 20 ng / ml enzyme in Tris-HCI buffer pH 7.2 for 15 minutes at 25 °C. The reaction is initiated by addition of 100 nM fluorescein lableled cAMP for another 30 minutes incubation period and terminated by addition of IMAP binding solution. IMAP complexes with phosphate groups on nucleotide monophosphate generated from cyclic nucleotides through PDE activity. Determination of the amount of complexformed is read spectrofluorimetrically at 470 nm / 525 nm. Compounds are screened at 10 pM. If binding is positive, additional concentrations are tested in order to calculate an IC50.

[0198] The Phosphodiesterase PDE3A (Human) assay is conducted in accordance with Hambleton, R., Krall, J., Tikishvili, E., Honeggar, M., Ahmad, F., Manganiello, V. C. and Movsesian, M. A. "Isoforms of cyclic nucleotide phosphodiesterase PDE3 and their contribution to cAMP hydrolytic activity in subcellular fractions of human myocardium", J. Biol. Chem. 280(47): 39168-39174, 2005, as shown below and in Table 1:■ 152300 Phosphodiesterase PDE3ASource: Human recombinant insect Sf9 cells Pre-Incub. Tlme / Temp: 15 minutes @ 25°CSubstrate: 0.10 pM FAM-cAMP Incubation Tlme / Temp: 30 minutes @ 25°CVehicle: 1.00% DMSO Incubation Buffer: 10 mM Tris-HCI, pH 7.2, 10 mM MgClz,0 05% NaNs, 0 1% phosphate-free BSASignificance Grit.: >50% of max stimulation or inhibition Quantitation Method: Quantitation of Fluorescein-AMP-IMAPTable 1. Reference data:*lndicates standard reference agent used. c) Transporter dopamine (DAT) assay:

[0199] Human recombinant dopamine transporters expressed in CHO-S cells are used in modified Tris-HCI buffer pH 7.4. A 0.4 pg aliquot is incubated with 0.15 nM [125l] RTI-55 for 3 hours at 4 °C. Non-specific binding is estimated in the presence of 10 pM nomifensine. Transporter are filtered and washed, the filters are then counted to determine [125l]RTI-55 specifically bound. Compounds are screened at 10 pM.

[0200] The Transporter dopamine (DAT) assay is conducted in accordance with the particulars as shown below and Table 2:■ 220320 Transporter, Dopamine (DAT)Table 2. Reference data:*lndicates the standard reference agent used.

[0201] The results of these experiments is summarized in Table 3.Table 3.

[0202] It is demonstrated by the above results that the tested example compound, compound 1, of the present invention exhibits high antibacterial activity against 5. aureus bacteria while being less prone to interact with PDE3 and DAT receptors than other Fabl inhibitors within the same compound family. Therefore, these results show that compound 1 is better tolerated than other, structurally closely related Fabl inhibitors.

[0203] Pharmacological Example 2: Plasma concentration profiles of compound 1 and compound 2 in the Staphylococcus aureus neutropenic murine thigh model

[0204] Neutropenic female ICR mice were inoculated intramuscularly (IM) with 5. aureus strain ATCC 29213. Compound 2 formulated in 5% dextrose was administered as single oral doses of 10, 20, 40 and 80 mg / kg (compound 1 equivalent concentrations), 2 hours after infection in volumes of 10 mL / kg / . Animals were sacrificed 0.083, 0.25, 0,5, 1, 2, 4, 8, and 24 hours post-dose and blood collected by cardiac puncture. Blood was drawn into tubes coated with K2EDTA, mixed and centrifuged at 2,500 xg for 15 minutes at 4°C. Plasma were transferred to LoBind polypropylene tubes containing a ratio of sample mouse plasma volume to human plasma (with protease inhibitor; complete™ protease inhibitor cocktail) volume of 1:1. Compound 2 and compound 1 concentrations were determined using LC-MS / MS.

[0205] Figure 1 shows the plasma concentration of compound 1 and compound 2 in relation to the time that has passed since their administration. These results show thatwhen dosing animals with compound 2, which is the prodrug of compound 1, systemic exposure of compound 1 is substantially higher than that of compound 2.

[0206] Pharmacological Example 3: In vivo efficacy of compound 1 (dosed as compound 2) in a Staphylococcus aureus neutropenic murine thigh infection model

[0207] The 5. aureus neutropenic murine thigh infection model was performed essentially as described in Lepak, A. J., Zhao, M. & Andes, D. R. Comparative Pharmacodynamics of Telavancin and Vancomycin in the Neutropenic Murine Thigh and Lung Infection Models against Staphylococcus aureus. Antimicrob Agents Chemother 61, doi:10.1128 / aac.00281-17 (2017). Female ICR mice were rendered neutropenic via intraperitoneal injection of cyclophosphamide four days before infection (150 mg / kg) and again one day before infection (100 mg / kg). Animals were anesthetized with isoflurane (3-5%) then inoculated intramuscularly with 5. aureus ATCC 29213 (~1 x 105CFU / mouse). Animals received compound 2 (in 5% dextrose) was administered via oral gavage twice daily. For control, animals received linezolid three times per day or 5% dextrose as a vehicle controls two times per day, by oral gavage. Animals were sacrificed via CO2 asphyxiation at either 2, 26 or 50 hours post infection. Thigh tissues were harvested and homogenized and bacterial counts were assessed by plating serial 10-fold dilutions and agar plates. The results are shown in Figure 2. These results show that compound 1 (dosed as compound 2) can achieve comparable reduction in CFU as linezolid at lower daily dosages.

Claims

Claims1. A compound for use in a method of treating a bacterial infection selected from infections associated with a Staphylococcal species in a patient, wherein the compound is represented by Formula I:Formula I or a pharmaceutically acceptable salt or prodrug or solvate thereof.

2. The compound for use according to claim 1, which is represented by Formula II:wherein X is selected from the group consisting of H and a pharmaceutically acceptable cation such as Na+or K+.

3. The compound for use according to claim 2, wherein the pharmaceutically acceptable cation is Na+.

4. The compound for use according to any one of claims 1 to 3, wherein the bacterial infection is associated with Staphylococcus aureus, preferably methicillin-resistant Staphylococcus aureus.

5. The compound for use according to any one of claims 1 to 4, wherein the compound is administered at a dosage of 40 mg / day to 3000 mg / day.

6. The compound for use according to any one of claims 1 to 5, wherein the compound is administered orally or intravenously.

7. The compound for use according to any one of claims 1 to 6, wherein the patient is a human.

8. The compound for use according to any one of claims 1 to 7, wherein the method comprises administration of one or more further antibiotics or antibacterial agents.

9. The compound for use according to any one of claims 1 to 8, wherein the bacterial infection is selected from the group consisting of acute bacterial skin and skin structure infections, bone and joint infections and diabetic foot infections.

10. A method of treatinga bacterial infection selected from infections associated with a Staphylococcal species, comprising administering to a patient in need thereof a pharmaceutically effective amount of a compound represented by Formula I:Formula I or a pharmaceutically acceptable salt or prodrug or solvate thereof, wherein preferably the method is specified in any one of claims 2 to 9.

11. A pharmaceutical composition for use in a method of treating a bacterial infection selected from infections associated with a Staphylococcal species in a patient, wherein the composition comprises the compound specified in any one of claims 1 to 9 and a pharmaceutically acceptable excipient.

12. A compound represented by the following Formula II:wherein X is a pharmaceutically acceptable cation such as Na+or K+.

13. A compound according to claim 12, wherein the pharmaceutically acceptable cation is Na+.

14. A pharmaceutical composition comprising the compound specified in claim 12 or 13 and a pharmaceutically acceptable excipient.

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