Aztreonam derivative

Abstract

Novel Compound The present invention relates to a pseudopaline analogue-aztreonam conjugate. The present invention further relates to salts, solvates and prodrugs of such compounds, to pharmaceutical compositions comprising such compounds, and to the use of such compounds in the treatment and prevention of medical disorders and diseases, most especially P. aeruginosa bacteria infections.

Description

[0001] AZTREONAM DERIVATIVE

[0002] Field of the Invention

[0003] The present invention relates to a pseudopaline analogue-aztreonam conjugate. The present invention further relates to salts, solvates and prodrugs of such compounds, to pharmaceutical compositions comprising such compounds, and to the use of such compounds in the treatment and prevention of medical disorders and diseases, most especially P. aeruginosa bacteria infections. Background

[0004] The increase and spread of antimicrobial resistance in bacteria is now a global public health problem. In contrast, only a few new antibiotics have been introduced since the 1970s. Metal acquisition is a vital microbial process in metal-scarce environments.

[0005] Staphylopine was discovered and functionally evaluated as a novel type of metallophore that Staphylococcus aureus employs to acquire multiple divalent transition metals. Importantly, homologous biosynthetic enzymes are also encoded by other microbial pathogens, such as the Gram-negative bacteria Pseudomonas aeruginosa. Indeed, a staphylopine like metallophore, named pseudopaline, was produced by P. aeruginosa for harvesting divalent metals.

[0006] Synthetic pseudopaline could promote the import of zinc, cobalt, iron, and nickel under the metal depleted condition, whereas epi-pseudopaline could not, indicating the importance of the configuration of glutaric acid moiety. A pseudopaline analogue-aztreonam conjugate exhibited better antibacterial effect towards P. aeruginosa than aztreonam alone.

[0007] Since the metal acquisition by P. aeruginosa is crucial for its ability to infect the human and cause diseases, the compound of the present invention may represent a potential treatment against P. aeruginosa bacterial infection.

[0008] Summary of the Invention

[0009] A first aspect of the invention provides a compound of formula (I):

[0010]

[0011] A second aspect of the invention provides a pharmaceutically acceptable salt, solvate or prodrug of any compound of the first aspect of the invention.

[0012] The compound of the present invention can be used both in their free base form and their acid addition salt form. For the purposes of this invention, a “salt” of a compound of the present invention includes an acid addition salt. Acid addition salts are preferably pharmaceutically acceptable, non-toxic addition salts with suitable acids, including but not limited to inorganic acids such as hydrohalogenic acids (for example, hydrofluoric, hydrochloric, hydrobromic or hydroiodic acid) or other inorganic acids (for example, nitric, perchloric, sulfuric or phosphoric acid); or organic acids such as organic carboxylic acids (for example, propionic, butyric, glycolic, lactic, mandelic, citric, acetic, benzoic, salicylic, succinic, malic or hydroxysuccinic, tartaric, fumaric, maleic, hydroxymaleic, mucic or galactaric, gluconic, pantothenic or pamoic acid), organic sulfonic acids (for example, methanesulfonic, trifluoromethanesulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, benzenesulfonic, toluene-p-sulfonic, naphthalene-2-sulfonic or camphorsulfonic acid) or amino acids (for example, ornithinic, glutamic or aspartic acid). The acid addition salt may be a mono-, di-, tri- or multi-acid addition salt. A preferred salt is a hydrohalogenic, sulfuric, phosphoric or organic acid addition salt. A preferred salt is a hydrochloric acid addition salt.

[0013] The compounds of the present invention can also be used both, in their free acid form and their salt form. For the purposes of this invention, a “salt” of a compound of the present invention includes one formed between a protic acid functionality (such as a carboxylic acid group) of a compound of the present invention and a suitable cation. Suitable cations include, but are not limited to lithium, sodium, potassium, magnesium, calcium and ammonium. The salt may be a mono-, di-, tri- or multi-salt. Preferably the salt is a mono- or di-lithium, sodium, potassium, magnesium, calcium or ammonium salt. More preferably the salt is a mono- or di-sodium salt or a mono- or dipotassium salt.

[0014] Preferably any salt is a pharmaceutically acceptable non-toxic salt. However, in addition to pharmaceutically acceptable salts, other salts are included in the present invention, since they have potential to serve as intermediates in the purification or preparation of other, for example, pharmaceutically acceptable salts, or are useful for identification, characterisation or purification of the free acid or base.

[0015] The compounds and / or salts of the present invention maybe anhydrous or in the form of a hydrate (e.g. a hemihydrate, monohydrate, dihydrate or trihydrate) or other solvate. Such solvates may be formed with common organic solvents, including but not limited to, alcoholic solvents e.g. methanol, ethanol or isopropanol.

[0016] In some embodiments of the present invention, therapeutically inactive prodrugs are provided. Prodrugs are compounds which, when administered to a subject such as a human, are converted in whole or in part to a compound of the invention. In most embodiments, the prodrugs are pharmacologically inert chemical derivatives that can be converted in vivo to the active drug molecules to exert a therapeutic effect. Any of the compounds described herein can be administered as a prodrug to increase the activity, bioavailability, or stability of the compound or to otherwise alter the properties of the compound. Typical examples of prodrugs include compounds that have biologically labile protecting groups on a functional moiety of the active compound. Prodrugs include, but are not limited to, compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, and / or dephosphorylated to produce the active compound. The present invention also encompasses salts and solvates of such prodrugs as described above.

[0017] The compounds, salts, solvates and prodrugs of the present invention may contain at least one chiral centre. The compounds, salts, solvates and prodrugs may therefore exist in at least two isomeric forms. The present invention encompasses racemic mixtures of the compounds, salts, solvates and prodrugs of the present invention as well as enantiomerically enriched and substantially enantiomerically pure isomers. For the purposes of this invention, a “substantially enantiomerically pure” isomer of a compound comprises less than 5% of other isomers of the same compound, more typically less than 2%, and most typically less than 0.5% by weight.

[0018] The compounds, salts, solvates and prodrugs of the present invention may contain any stable isotope including, but not limited to12C,13C,JH,2H (D),14N,15N,160,170,180,19F and127I, and any radioisotope including, but not limited tonC,14C,3H (T),13N,150,18F,1231,1241,1 31 and131I.

[0019] The compounds, salts, solvates and prodrugs of the present invention may be in any polymorphic or amorphous form. A third aspect of the invention provides a pharmaceutical composition comprising a compound of the first aspect of the invention, or a pharmaceutically acceptable salt, solvate or prodrug of the second aspect of the invention, and a pharmaceutically acceptable excipient. Conventional procedures for the selection and preparation of suitable pharmaceutical formulations are described in, for example, “Aulton’s Pharmaceutics - The Design and Manufacture of Medicines”, M. E. Aulton and K. M. G. Taylor, Churchill Livingstone Elsevier, 4thEd., 2013. Pharmaceutically acceptable excipients including adjuvants, diluents or carriers that may be used in the pharmaceutical compositions of the invention are those conventionally employed in the field of pharmaceutical formulation, and include, but are not limited to, sugars, sugar alcohols, starches, ion exchangers, alumina, aluminium stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycerine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. A fourth aspect of the invention provides a compound of the first aspect of the invention, or a pharmaceutically acceptable salt, solvate or prodrug of the second aspect of the invention, or a pharmaceutical composition of the third aspect of the invention, for use in medicine, and / or for use in the treatment or prevention of a disease, disorder or condition. Typically, the use comprises the administration of the compound, salt, solvate, prodrug or pharmaceutical composition to a subject.

[0020] The term “treatment” as used herein refers equally to curative therapy, and ameliorating or palliative therapy. The term includes obtaining beneficial or desired physiological results, which may or may not be established clinically. Beneficial or desired clinical results include, but are not limited to, the alleviation of symptoms, the prevention of symptoms, the diminishment of extent of disease, the stabilisation (i.e., not worsening) of a condition, the delay or slowing of progression / worsening of a condition / symptoms, the amelioration or palliation of the condition / symptoms, and remission (whether partial or total), whether detectable or undetectable. The term “palliation”, and variations thereof, as used herein, means that the extent and / or undesirable manifestations of a physiological condition or symptom are lessened and / or time course of the progression is slowed or lengthened, as compared to not administering a compound, salt, solvate, prodrug or pharmaceutical composition of the present invention. The term “prevention” as used herein in relation to a disease, disorder or condition, relates to prophylactic or preventative therapy, as well as therapy to reduce the risk of developing the disease, disorder or condition. The term “prevention” includes both the avoidance of occurrence of the disease, disorder or condition, and the delay in onset of the disease, disorder or condition. Any statistically significant (p < 0.05) avoidance of occurrence, delay in onset or reduction in risk as measured by a controlled clinical trial may be deemed a prevention of the disease, disorder or condition. Subjects amenable to prevention include those at heightened risk of a disease, disorder or condition as identified by genetic or biochemical markers. Typically, the genetic or biochemical markers are appropriate to the disease, disorder or condition under consideration and may include for example, inflammatory biomarkers such as C-reactive protein (CRP) and monocyte chemoattractant protein 1 (MCP-i) in the case of inflammation; total cholesterol, triglycerides, insulin resistance and C-peptide in the case of NAFLD and NASH; and more generally IL1P and IL18 in the case of a disease, disorder or condition responsive to NLRP3 inhibition. A fifth aspect of the invention provides the use of a compound of the first aspect, or a pharmaceutically effective salt, solvate or prodrug of the second aspect, in the manufacture of a medicament for the treatment or prevention of a disease, disorder or condition. Typically, the treatment or prevention comprises the administration of the compound, salt, solvate, prodrug or pharmaceutical composition to a subject.

[0021] A sixth aspect of the invention provides a method of treatment or prevention of a disease, disorder or condition, the method comprising the step of administering an effective amount of a compound of the first aspect, or a pharmaceutically acceptable salt, solvate or prodrug of the second aspect, or a pharmaceutical composition of the third aspect, to thereby treat or prevent the disease, disorder or condition. Typically, the administration is to a subject in need thereof.

[0022] An seventh aspect of the invention provides a compound of the first aspect of the invention, or a pharmaceutically acceptable salt, solvate or prodrug of the second aspect of the invention, or a pharmaceutical composition of the third aspect of the invention, for use in the treatment or prevention of a disease, disorder or condition in an individual, wherein the individual has a P. aeruginosa bacterial infection. Typically, the use comprises the administration of the compound, salt, solvate, prodrug or pharmaceutical composition to the individual.

[0023] Any of the medicaments employed in the present invention can be administered by oral, parenteral (including intravenous, subcutaneous, intramuscular, intradermal, intratracheal, intraperitoneal, intraarticular, intracranial and epidural), airway (aerosol), rectal, vaginal or topical (including transdermal, buccal, mucosal and sublingual) administration.

[0024] Typically, the mode of administration selected is that most appropriate to the disorder or disease to be treated or prevented.

[0025] For oral administration, the compounds, salts, solvates or prodrugs of the present invention will generally be provided in the form of tablets, capsules, hard or soft gelatine capsules, caplets, troches or lozenges, as a powder or granules, or as an aqueous solution, suspension or dispersion. Tablets for oral use may include the active ingredient mixed with pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavouring agents, colouring agents and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose. Corn starch and alginic acid are suitable disintegrating agents. Binding agents may include starch and gelatine. The lubricating agent, if present, may be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material, such as glyceryl monostearate or glyceryl distearate, to delay absorption in the gastrointestinal tract. Tablets may also be effervescent and / or dissolving tablets.

[0026] Capsules for oral use include hard gelatine capsules in which the active ingredient is mixed with a solid diluent, and soft gelatine capsules wherein the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin or olive oil.

[0027] Powders or granules for oral use may be provided in sachets or tubs. Aqueous solutions, suspensions or dispersions may be prepared by the addition of water to powders, granules or tablets. Any form suitable for oral administration may optionally include sweetening agents such as sugar, flavouring agents, colouring agents and / or preservatives.

[0028] Formulations for rectal administration maybe presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate.

[0029] Formulations suitable for vaginal administration maybe presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate. For parenteral use, the compounds, salts, solvates or prodrugs of the present invention will generally be provided in a sterile aqueous solution or suspension, buffered to an appropriate pH and isotonicity. Suitable aqueous vehicles include Ringer’s solution and isotonic sodium chloride or glucose. Aqueous suspensions according to the invention may include suspending agents such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone and gum tragacanth, and a wetting agent such as lecithin. Suitable preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate. The compounds of the invention may also be presented as liposome formulations.

[0030] For transdermal and other topical administration, the compounds, salts, solvates or prodrugs of the invention will generally be provided in the form of ointments, cataplasms (poultices), pastes, powders, dressings, creams, plasters or patches.

[0031] Suitable suspensions and solutions can be used in inhalers for airway (aerosol) administration.

[0032] The dose of the compounds, salts, solvates or prodrugs of the present invention will, of course, vary with the disorder or disease to be treated or prevented. In general, a suitable dose will be in the range of o.oi to 500 mg per kilogram body weight of the recipient per day. The desired dose maybe presented at an appropriate interval such as once every other day, once a day, twice a day, three times a day or four times a day. The desired dose may be administered in unit dosage form, for example, containing 1 mg to 50 g of active ingredient per unit dosage form.

[0033] For the avoidance of doubt, insofar as is practicable any embodiment of a given aspect of the present invention may occur in combination with any other embodiment of the same aspect of the present invention. In addition, insofar as is practicable it is to be understood that any preferred, typical or optional embodiment of any aspect of the present invention should also be considered as a preferred, typical or optional embodiment of any other aspect of the present invention.

[0034] Compound synthesis

[0035] Compound 4.29 can be successfully condensed with 4.16 and the resulting imine reacted with 4.15. The target product 4.30 was obtained in 61% yield with a diastereoselectivity of 15 to 1.

[0036] The amine of 4.30 is protected by trifluoroacetylation by treatment with TFAA and pyridine. Saturated sodium bicarbonate solution is then added to the reaction mixture, from withthe target product 4.31 is isolated.

[0037] Next, ozonolysis of the alkene, reductive amination of the resulting aldehyde with a protected histidine derivate and two-step deprotection led to the pseudopaline derivative 4.24 (Scheme 1).

[0038] Compound 4.16 (1 eq) and anhydrous sodium sulfate (2 g) were combined in dichloromethane (40 mL). Compound 4.29 (1.1 eq) in dichloromethane (10 mL) was added at o °C and the mixture stirred at room temperature for 0.5 hour. The mixture was cooled to -78 °C, treated with tin tetrachloride (1.2 eq. 1 M in dichloro methane) and compound 4.15 (2 eq), and stirred at o °C for 4 hours. The mixture was quenched with sodium bicarbonate solution until alkaline, water (50 ml) added, and the product extracted with methylene chloride (3x100ml), washed with saturated brine (30 ml). dried and concentrated, and the residue was further purified by silica gel column chromatography (petroleum ether: ethyl acetate = 7:1) to obtain a compound 4.30 at 61% yield, d.r.=i5:i, as a colorless oily compound.

[0039] 1H NMR (400 MHz, CDCI3) 8 7.39 - 7.28 (m, 15H), 5.99 (s, 1H), 5.73 - 5.60 (m, 1H), 5.18 - 5.05 (m, 8H), 3.48 - 3.29 (m, 4H), 2.58 - 2.47 (m, 1H), 2.40 - 2.27 (m, 1H),

[0040] 2.04 - 1.92 (m, 1H), 1.73 (s, 1H), 1.64 - 1.48 (m, 1H);

[0041] 13C NMR (101 MHz, CDCI3) 8 174.5, 173-6, 156.5, 136.9, 135-4, 135-4, 133-1, 128.6,

[0042] 128.6, 128.4, 128.4, 128.4, 128.3, 127.9, 127.9, 118.9, 66.8, 66.8, 66.3, 58.8, 58.0, 38.1, 37-6, 32.7; [a]D22 -39.4 (c 1, CHCI3)

[0043] Compound 4.31

[0044] Compound 4.30 (1 eq) was dissolved in dichloromethane (8 mL), and pyridine (8 eq) and trifluoroacetic anhydride (0.435 mL, 3.1 mmol, 1.25 eq) were added at o°C, and the mixture stirred at room temperature overnight.

[0045] The reaction was quenched with sodium bicarbonate (10 mL) for 12 hours, diluted with ethyl acetate (30 mL).The organic phase was washed with 1 N hydrochloric acid (5 mL) and saturated saline (5 mL) and dried over anhydrous sodium sulfate. The mixture was concentrated and the residue purified by silica gel column chromatography (petroleum ether: ethyl acetate =10:1) to obtain the colorless oily compound 4.31. 1H NMR (400 MHz, CDCI3) 8 7.41 - 7.26 (m, 12H), 7.25 - 7.11 (m, 3H), 5.88 - 5.71 (m, O.33H), 5-66 - 5.50 (m, 0.63H), 5.18 - 5.00 (m, 7H), 4.66 - 4.47 (m, 2H), 4.43 (dd, J = 8.8, 2.6 Hz, 0.62H), 4.31 (t, J = 6.9 Hz, 0.36H), 3.42 (dd, J = 12.5, 6.2 Hz, 1H), 3.26 - 3.11 (m, 0.70H), 3.10 - 3.00 (m, 0.37H), 2.78 - 2.59 (m, 1.60H), 2.54 - 2.40 (m, 0.55H), 2.20 - 2.08 (m, 1H), 1.96 - 1.85 (m, 0.85H). 13C NMR (126 MHz, CDCI3) 8 169.1, 168.4, 168.2, 167.8, 157.1, 156.8, 156.5, 136.4,

[0046] 135.2, 135.0, 134.8, 134.2, 130.7, 128.5, 128.5, 128.5, 128.4, 128.4, 128.4, 128.3, 128.2, 128.2, 128.1, 128.1, 128.0, 120.4, 118-7, 117-0, 114-8, 67-7, 67-7, 67-6, 67-3, 66-9, 66.8, 59-1, 57-1, 55-4, 39-4, 37-4, 34-7, 33-5, 31-5, 31-1-

[0047] HRMS (ESI): [M+H]+ calculated for C33H34F3N2O7: 627.2313, found: 627.2309; [a]D22 -35.0 (c 1, CHCI3)

[0048] Compound 4.33

[0049] Compound 4.31 (1.0 eq) was dissolved in dichloromethane (10 mL) and then cooled to -

[0050] 78 °C and treated with ozone for 5 minutes. The reaction was quenched with dimethyl sulfide (0.4 mL), concentrated, and the residue was subjected to silica gel column chromatography (petroleum ether: acetic acid ethyl ester = 10:1) to obtain colorless oily compound 4.32 (81%).

[0051] 1H NMR (400 MHz, CDCI3) 8 9.75 (s, 0.69H), 9.61 (s, 0.20H), 7.37 - 7.19 (m, 15H), 5.29 - 5.04 (m, 6H), 4.73 - 4.67 (m, 1.5H), 4.53 - 4.48 (brs, 0.26H), 4.32 (dd, J = 9.4, 3.9 Hz, 0.23H), 3.84 - 3.77 (m, 0.72H), 3.62 (dd, J = 18.6, 9.9 Hz, 0.23H), 2.89 (dd, J = 18.6, 4.2 Hz, 0.26H), 2.76 (brs, 0.45H), 2.66 - 2.58(111, 1H), 2.40 - 2.25 (m, 2.34H), 2.13 - 2.02 (m, 0.82H);

[0052] 13C NMR (101 MHz, CDCI3) 8 197.27, 197.17, 173.20, 171.34, 168.53, 168.02, 167.73, 158.06, 157-76, 157-47, 135-61, 135-36, 134-97, 134-82, 134-68, 128.67, 128.63, 128.59, 128.52, 128.50, 128.49, 128.45, 128.41, 128.39, 128.31, 128.10, 127.83, 116.90, 114.61, 68.08, 67.67, 67.64, 66.86, 66.66, 58.93, 44.61, 30.67, 29.72, 29.65, 25.35, 24.95;

[0053] IR (neat) vmax 2955, 1739, 1697, 1498, 1456, 1386, 1194, 1150, 1084 cm-1;

[0054] HRMS (ESI): [M+H]+ calculated for C32H31F3NO8: 614.1996, found: 614.1988;

[0055] [a]D24 -39 (c 2.5, CHCI3);

[0056] Rf = 0.32 (PE / EA = 5 / I).

[0057] Compound 4.16 (1.5 eq), sodium acetate (5 eq), molecular sieves (70 mg) were combined in isopropanol (2 mL), and 4.31 (1 eq dissolved in 2 mL methanol ) added at room temperature. After to minutes, sodium cyanoborohydride (5 eq) was added at o °C, and the mixture stirred at room temperature for 2 hours.

[0058] The reaction mixture was concentrated to dryness, and ethyl acetate (20 mL) and water (5 mL) added. The organic phase was washed with saturated brine (5 mL), then dried, concentrated, and purified by high performance liquid phase to obtain colorless oily compound 4.33 (73%).

[0059] 1H NMR (400 MHz, CDCI3) 88.12 (s, 0.76H), 8.06 (s, 0.23H), 7.42 - 7.28 (m, 20H),

[0060] 7.19 - 7.09 (m, 10H), 6.94 (s, 0.78H), 6.80 (s, 0.22H), 5.70 (s, 0.52H), 5.55 (s, 0.20H), 5-13 - 4-90 (m, 6H), 4.59 (t, J = 7.2 Hz, 1H), 4.55 - 4.47 (m, 1H), 4.36 - 4.23 (m, 1H),

[0061] 3.69 (s, 3H), 3.66 -3.60 (m, 1H), 3.60 - 3.53 (m, 1H), 3.53 - 3.31 (m, 3H), 3.30 - 3.08 (m, 2H), 2.94 -2.68 (m, 1H), 2.33 -2.13 (m, 1H), 2.08 -1.87 (m, 1H).

[0062] 13C NMR (101 MHz, CDCI3) 8 168.8, 167.1, 161.9, 161.6, 156.8, 139.5, 139-4, 136.4, 136.0, 134.6, 134.6, 129.4, 129.2, 128.8, 128.5, 128.5, 128.4, 128.3, 128.0, 127.9, 127.8, 127.5, 122.8, 117.4, 114-5, 99-9, 78.8, 77.2, 67.9, 67.4, 66.6, 58.9, 53.5, 45.5, 37.4, 25.2.

[0063] HRMS (ESI): [M+H]+ calculated for C58H57F3N5O7: 1024.4103, found: 1024.4085;

[0064] [a]D22 8.1 (c 1, CHCI3).

[0065] Compound 4.24

[0066] Compound 4.33 (1 eq) and anisole (20 eq) were dissolved in dichloromethane (2 mL), , cooled to o °C, and trifluoromethanesulfonic acid (15 eq) added.

[0067] The mixture was stirred at o°C for 0.5 hours, then raised to room temperature, stirred for 2 hours, cooled to o°C, and sodium bicarbonate (30 eq) in water (10 mL) added and the mixture stirred for another 0.5 hour, and wash with dichloro methane (5 mLx3).

[0068] The aqueous phase was concentrated to obtain crude product. The crude product was dissolved in water (1.5 mL) and methanol (1.5 mL), and potassium carbonate (6 eq) was stirred at 66 °C for 24 h, then concentrated in vacuo, and the resulting solid was purified to give compound 4.24 (71 % as a white solid). 1H NMR (400 MHz, D2O) 8 7.75 (d, J = 20.7 Hz, 1H), 7.00 (s, 1H), 3.83 (t, J = 6.0 Hz, 1H), 3.28 - 2.99 (m, 8H), 2.02 - 1.75 (m, 4H);

[0069] 13C NMR (126 MHz, D2O) 8 180.3, 173-7, 136.1, 131.6, 117.2, 62.0, 61.6, 60.8, 45.5, 37- , 30.0, 28.9, 27.4;

[0070] HRMS (ESI): [M+H]+ calculated for C14H24N5O6: 358.1721, found: 358.1724;

[0071] [a]D22 -6.7 (c 0.2, H20)O

[0072] Compound 4.62

[0073] Aztreonam (1 eq), EDCI (1.2 eq) and NHS (1 eq) were dissolved in DMF (1 mL) and DIPEA (4 eq) and stirred for 12 hours. Compound 4.24 (0.2 eq) in PBS buffer (1 mL) was then added and the mixture stirred at room temperature for 1 hour. Compound4.62 (38%) was purified from the crude mixture.

[0074] 1H NMR (400 MHz, D2O) 8 8.64 (s, 1H), 7.38 (s, 1H), 7.13 (s, 1H), 4.68 (d, J = 2.6 Hz, 1H), 4.30 - 4.21 (m, 1H), 4.09 - 4.02 (m, 1H), 3.85 (t, J = 6.2 Hz, 2H), 3.58 - 3.49 (m,

[0075] 1H), 3-49 - 3-38 (m, 2H), 3.38 - 3.29 (m, 3H), 2.35 (dd, J = 14.2, 6.9 Hz, 2H), 2.25 - 2.11 (m, 2H), 1.56 (d, J = 6.2 Hz, 3H), 1.51 (d, J = 4.2 Hz, 6H);

[0076] 13C NMR (151 MHz, D2O) 8 176.6, 172.9, 171.5, 170.8, 170.7, 164.6, 161.6, 144.1, 134.0, 130.4, 126.6, 117.9, 111-9, 85.2, 61.1, 60.2, 59.4, 59.2, 58.4, 43.9, 35.3, 29.4, 26.7, 24.7, 23.4, 23.1, 16.8;

[0077] HRMS (ESI): [M-H]- calculated for C27H37N1OO13S2: 773.1988, found: 773.1984

[0078] Biological studies

[0079] The compound of the invention has been evaluated using the following protocol. The Minimal Inhibitory Concentration (MIC) of compounds was determined by broth microdilution assay according to M07-Ed.11 and Mioo-Ed.32 Clinical Laboratory and Standard Institute (CLSI) guidelines. Solutions of test compounds and antibiotic reference compounds (aztreonam, aztreonam / avibactam, cefiderocol, and ciprofloxacin) were prepared at 50X the final desired concentration in either 100% dimethyl sulfoxide (DMSO, Sigma Aldrich) or water. The test compounds, along with reference compounds, were serially diluted 2-fold across the 96-well plates (Greiner Bio-One): compounds ranged from 64 pg / mL to 0.06 pg / mL or from 32pg / mL to 0.03 pg / mL, standards ranged from 32 pg / mL to 0.03 pg / mL. Bacteria were cultured overnight in Tryptone Soya agar (TSA, Thermo Fisher Diagnostics Spa). The inoculum was prepared by making a direct saline suspension of isolated colonies selected from agar plates. It was adjusted to achieve a turbidity equivalent to the 0.5 McFarland standard. The suspension was then diluted 1:200 in Cation-Adjusted Mueller Hinton II broth (CAMHB, BBL BD). To deplete CAMHB broth of iron or zinc, 100g of Chelex 100 resin (Bio-Rad) was added to 1L for 2 hours before filtering out. To make iron-depleted broth, Zn2+(o.5-i.omg / L final concentration), Mg2+ (io-i2.5mg / L final concentration), and Ca2+ (20-25mg / L final concentration) were supplemented. To make zinc-depleted broth Fe2+(o.25mg / L final concentration), Mg2+ (io-i2.5mg / L final concentration), and Ca2+ (20-25mg / L final concentration) were supplemented. The ion-depleted medium was adjusted to 7.2 pH, and filter-sterilized. Every plate tested included a positive control of just the bacteria and a negative control of only the medium. All the plates were covered and incubated at 35±2 °C in ambient air for 20 to 24 hours. MICs were determined visually, with the MIC defined as the lowest concentration at which no growth was visible after incubation.

Claims

Claims1. A compound of formula (I):

2. A pharmaceutically acceptable salt, solvate or prodrug of a compound as claimed in claim 1.

3. A pharmaceutical composition comprising a compound as claimed in claim 1, or a pharmaceutically acceptable salt, solvate or prodrug as claimed in claim 2, and a pharmaceutically acceptable excipient.

4. A compound as claimed in claim 1, or a pharmaceutically acceptable salt, solvate or prodrug as claimed in claim 2, or a pharmaceutical composition as claimed in claim3, for use in medicine.

5. A compound, pharmaceutically acceptable salt, solvate, prodrug or pharmaceutical composition as claimed in any claims 1 to 4, for use in the treatment or prevention of a P. aeruginosa bacteria infection.

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