JP2006515306A - Oligosaccharides and conjugates thereof for the treatment of Pseudomonas bacterial infections - Google Patents

Abstract

In connection with Pseudomonas bacteria, compositions and methods are provided. The compositions and methods can be used for the diagnosis and treatment of medical conditions associated with infection by Pseudomonas bacteria. Such infectious diseases include Pseudomonas aeruginosa in the lungs of patients with cystic fibrosis. Compounds useful in the present methods can be conjugated to therapeutic agents. Pseudomonas bacteria can be inhibited by blocking colonization, by preventing the growth of the bacteria, or by killing the bacteria.

Description

(Background of the Invention)
(Field of Invention)
The present invention generally relates to the diagnosis of disease in warm-blooded animals (eg, humans) involved in infection and colonization by Pseudomonas bacteria, including Pseudomonas aeruginosa, in the lungs of patients with cystic fibrosis. It relates to compounds, compositions and methods for treatment. The present invention more specifically relates to the use of one or more compounds to selectively bind to Pseudomonas bacteria. These compounds are useful for diagnosis and / or therapeutic intervention of colonization of Pseudomonas bacteria, or can be conjugated to drugs to target Pseudomonas bacteria and effectively block or kill Pseudomonas bacteria.

(Description of related technology)
Pseudomonas infections can occur in a variety of medical conditions and can be life threatening. Pseudomonas is an opportunistic bacterium. Examples of individuals at risk include cystic fibrosis patients and burn patients. Cystic fibrosis is described below as a representative example of a medical condition that may include infection with Pseudomonas bacteria.

  Cystic fibrosis (CF) is the most common lethal genetic disease among the Caucasian population. CF is caused by mutations in the gene encoding a cystic fibrosis transmembrane conductance regulator (CFTR) and acts as a chloride channel. Genetic mutations in CFTR that alter ion migration also affect N-glucosylation of CFTR and other cell surface molecules. All exocrine glands of the patient are affected; however, the lung is a major part of morbidity and mortality. General changes in glucosylation result in an increase in Lewis fucosylation and a decrease in sialylation. Salivary and respiratory mucins from CF patients also contain higher levels of Lewis-type oligosaccharides, including sialylated and sulfated Lewis x / a structures.

  The main cause of morbidity and mortality in CF patients is chronic lung colonization by this bacterium, Pseudomonas aeruginosa, which causes a marked pulmonary infection with a strong neutrophilic inflammatory response leading to lung destruction and death. Arise. P. Colonization by aeruginosa is initiated by the binding of ciliate and flagellar lectins on this bacterium to a Lewis-type sugar chain structure on the lung cell surface. These lectins, known as PA-IL and PA-IIL, represent potential molecular targets that bind with high affinity to these oligosaccharide structures and prevent the initial steps of bacterial colonization. Patients who are not well colonized by this bacterium maintain a good long-term prognosis. Due to the difficulties in current approaches in the art for preventing colonization in individuals by Pseudomonas bacteria, compounds, compositions and methods need to be improved.

(Simple Summary of Invention)
Briefly, the present invention relates to compounds, compositions and methods for utilizing lectins expressed on Pseudomonas bacteria for the detection of Pseudomonas bacteria, as well as diseases involving Pseudomonas bacteria (including human diseases). Provide diagnosis and treatment. For example, P.I. A Lewis-structured sugar mimetic with high affinity binding to a lectin on aeruginosa has a beneficial therapeutic effect on CF patients. In addition, these sugar mimetics can be conjugated with strong antibiotics, for example, to increase efficacy and reduce dosage, thereby avoiding the known harmful side effects of these powerful antibiotics Can be done. Considering that these binding moieties are critical to the colonization and pathogenicity of this bacterium, mutations in this target that are resistant to this conjugate therapy are non-pathogenic forms of this bacterium. Must produce.

  One embodiment of the present invention provides a method of inhibiting Pseudomonas bacteria in warm-blooded animals, wherein the method is effective for inhibiting compounds comprising the compounds described in FIG. 1 or FIG. 2 to inhibit the bacteria. A step of administering to the animal in an amount.

  In another embodiment, the present invention provides a conjugate comprising a therapeutic agent conjugated to a compound described in FIG. 1 or FIG.

  In another embodiment, the present invention provides a method of detecting Pseudomonas bacteria, wherein a method includes a diagnostic agent and a sample bound to a compound comprising a compound described in FIG. 1 or FIG. 2 in the sample. If present, contacting the compound under conditions sufficient to bind to the bacterium; and detecting the agent present in the sample, wherein the agent in the sample The presence of comprises a step indicating the presence of Pseudomonas bacteria.

  In another embodiment, the present invention provides a method of immobilizing Pseudomonas bacteria on a solid support, the method comprising, on a solid support, a sample containing Pseudomonas bacteria under conditions sufficient to bind. Contacting with a compound comprising an immobilized compound as described in FIG. 1 or FIG. 2; and separating the sample from the solid support.

  In other embodiments, the compounds and conjugates described herein can be used in the preparation of a medicament for the inhibition of Pseudomonas bacteria.

  These and other aspects of the invention will be apparent upon reference to the following detailed description and attached drawings. All references disclosed herein are incorporated by reference in their entirety as if each was individually incorporated.

(Detailed description of the invention)
As described above, the present invention relates to P.I. Provided are compounds and compositions that can be used in the diagnosis and treatment of disease in combination with aeruginosa.

(Sugar mimetic compound)
As used herein, the term “sugar mimetic compound” refers to P.I. A molecule that specifically binds to aeruginosa. The structure of the sugar mimetic compound encompassed by the present invention is shown in FIGS. 1 and 2, and also this compound does not contain a mimic for sialic acid shown as a terminal cyclohexyl lactate moiety. Includes compounds disclosed herein. This is accomplished, for example, by removing steps that include the addition of intermediate E in certain reaction schemes. All compounds (or conjugates thereof) useful in the present invention include physiologically acceptable salts thereof.

  For certain embodiments, it may also be beneficial to conjugate a diagnostic or therapeutic agent, such as a drug, with a glycomimetic compound to form a conjugate (the linkage is covalent). As used herein, the term “therapeutic agent” refers to a warm-blooded animal (eg, such as a human) to prevent or treat a disease or other undesired condition, or to promote therapeutic success. Any biologically active agent intended for administration to a mammal. Therapeutic agents include antibiotics, hormones, growth factors, proteins, peptides, genes, non-viral vectors and other compounds.

(Glucose mimetic compound formulation)
The sugar mimetic compounds described herein can be present in a pharmaceutical composition. The pharmaceutical composition comprises one or more sugar mimetic compounds in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. Such a composition can be a buffer (eg, neutral buffered saline or phosphate buffered saline), carbohydrate (eg, glucose, mannose, sucrose, or dextran), mannitol, protein, polypeptide, or It may contain amino acids such as glycine, antioxidants, chelating agents (eg EDTA or glutathione), adjuvants (eg aluminum hydroxide) and / or preservatives. In still other embodiments, the compositions of the invention can be formulated as lyophilizates. The compositions of the invention are formulated for any suitable mode of administration, including, for example, aerosol administration, topical administration, oral administration, nasal administration, intravenous administration, intracranial administration, intraperitoneal administration, subcutaneous administration, or intramuscular administration. Can be done.

  A pharmaceutical composition can also include one or more active agents, such as drugs that can alternatively be conjugated to a sugar mimetic compound or free within the composition. The binding of the drug to the glycomimetic compound can be covalent or non-covalent.

  The compositions described herein can be administered as part of a sustained release formulation (ie, a formulation such as a capsule or sponge that provides a slow release of the modulating agent after administration). Such formulations are generally prepared using well-known techniques and can be administered, for example, by oral, rectal or subcutaneous implantation, or implantation at the desired target site. Carriers for use within such formulations are biocompatible and may also be biodegradable; preferably the formulation provides a relatively constant level of controlled drug release. provide. The amount of glycomimetic compound included in the sustained release formulation depends on the site of implantation, release rate, and expected duration of release, and the nature of the condition being treated or prevented.

  The sugar mimetic compound is generally present in the pharmaceutical composition in a therapeutically effective amount. A therapeutically effective amount is an amount that provides a discernible benefit for the patient, such as a measurement of a condition associated with Pseudomonas infection or an observed response.

(How to use sugar mimetic compounds)
In general, the glycomimetic compounds described herein are used to achieve diagnostic and / or therapeutic results that result in diseases (eg, human diseases) that are associated with infection by Pseudomonas (eg, P. aeruginosa) bacteria. Can be used. Such diagnostic and / or therapeutic results can be achieved in vitro and / or in vivo in animals, preferably mammals such as humans. However, Pseudomonas (eg, P. aeruginosa) is ultimately contacted with a glycomimetic compound in an amount and time sufficient to achieve a recognizable diagnostic or therapeutic result. In the context of the present invention, treatment outcome is associated with, for example, prevention of lung infection. In some conditions, the outcome of treatment is inhibition of Pseudomonas (eg, P. aeruginosa) (eg, preventing the growth of such bacteria or killing the bacteria, or preventing colonization by the bacteria). Related). As used herein, therapeutic or therapeutic outcome includes treatment or prevention.

  The glycomimetic compounds of the invention can be administered in a manner appropriate to the disease to be treated or prevented. The appropriate dosage and the appropriate duration and frequency of administration can be determined by factors such as the patient's condition, the type and severity of the patient's disease and the method of administration. In general, an appropriate dosage and treatment regimen provides the modulating agent in an amount sufficient to provide a treatment and / or prevention benefit. In a particularly preferred embodiment of the invention, the glycomimetic compound is administered daily at a dosage ranging from 0.001 mg / kg body weight to 1000 mg / kg body weight (more typically from 0.01 mg / kg to 1000 mg / kg). It can be administered in single or multiple dose regimens. Appropriate dosages can generally be determined using experimental models and / or clinical trials. In general, the use of the minimum dosage that is sufficient to provide effective therapy is preferred. Patients can generally be monitored for therapeutic efficacy using assays appropriate to the condition to be treated or prevented that are familiar to those skilled in the art.

  Glucose mimetic compounds can also be used to target substances to Pseudomonas bacteria (eg, P. aeruginosa). Such substances include therapeutic agents and diagnostic agents. Therapeutic agents can be demonstrated to alter the properties of molecules, viruses, viral components, cells, cellular components, or target cells, so as to treat or prevent disorders or to control the physiology of a patient It can be any other substance that can provide the benefit of. The therapeutic agent can also be a prodrug that results in an agent having biological activity in vivo. Molecules that can be therapeutic agents can be, for example, polypeptides, amino acids, nucleic acids, polynucleotides, steroids, polysaccharides, or inorganic compounds. Such molecules can function in any of a variety of ways including, for example, enzymes, enzyme inhibitors, hormones, receptors, antisense oligonucleotides, catalytic polynucleotides, antiviral agents, antitumor agents, anti-tumor agents, Bacterial agents, immunomodulators, and cytotoxic agents (eg, radionuclides such as iodine, bromine, lead, rhenium, homium, palladium, or copper). Diagnostic agents include imaging agents such as metals and radiopharmaceuticals (eg gallium, technetium, indium, strontium, iodine, barium, bromine and phosphorus containing compounds), contrast agents, dyes (eg fluorescent dyes or fluorophores) And enzymes that catalyze colorimetric or fluorometric reactions. In general, therapeutic and diagnostic agents can be conjugated to glycomimetic compounds using a variety of techniques, such as those described above. For targeting purposes, glycomimetic compounds can be administered to a patient as described herein.

  Glucose mimetic compounds can also be used in vitro (eg, in various well-known cell culture and cell separation methods). For example, a glycomimetic compound can be immobilized on a solid support for use in immobilizing Pseudomonas bacteria for screening, assay or growth in culture. (For example, linked to the internal surface of a tissue culture plate or other cell culture support). Such a connection can be made by any suitable technique, such as the method described above, and other standard techniques. Also, the use of glycomimetic compounds facilitates cell identification and in vitro sorting and may allow selection of such bacterial cells. Preferably, a glycomimetic compound for use in such a method is linked to a diagnostic agent that is a detectable marker. Suitable markers are well known in the art and include radionuclides, luminescent groups, fluorescent groups, enzymes, dyes, immunoglobulin constant domains, and biotin. In one preferred embodiment, a glycomimetic compound linked to a fluorescent marker such as fluorescein is contacted with cells and then analyzed by a fluorescence activated cell sorter (FACS).

  Such in vitro methods generally involve contacting a sample (eg, a biological preparation) with any one of the sugar mimetic compounds and detecting the compound in the sample. Is included. If desired, one or more washing steps can be added to the method. For example, the sample can be contacted with the sugar mimetic compound prior to detection of the compound, and then the sample can be washed (ie, contacted with a fluid and then removed the fluid and removed unbound sugar mimetic compound). To do). Alternatively or in addition, a washing step can be further added during this detection process. For example, if the glycomimetic compound possesses a marker (diagnostic agent) that can bind to a detectable substance, the sample is washed after contacting the sample with the detectable substance prior to the detection. It may be desirable to do so. As used herein, the phrase “detection of a compound (or drug) in this sample” is to detect this compound (or drug) where this compound (or drug) remains bound to this sample. Or detecting the compound (or drug) bound to the sample after the compound (or drug) has been separated from the sample.

  The following examples are presented for purposes of illustration and not for purposes of limitation.

Example 1
(Synthesis of 3 (Fig. 3))
(Formation of intermediate C :)
Compound A (5.00 g, 12.74 mmol) and B (4.50 g, 19.11 mmol) and NIS (3.58 g, 15.93 mmol) were dissolved in CH ₂ Cl ₂ (50 ml) and cooled to 0 ° C. To do. Of trifluoromethanesulfonic acid solution _(CH 2 Cl ₂ solution of 0.15 M), is added dropwise with stirring. When the solution changes color from orange to dark brown, the addition of TMS-OH is stopped. The solution is then washed with saturated NaHCO ₃ (30 ml) and the organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. The resulting syrup is purified by silica gel chromatography (hexane / ether, 1: 1) and used in the next step.

The compound obtained above is dissolved in THF (40 ml) and carbon-supported Pd (10%) (1/10 by mass) is added. The solution was degassed, to generate an atmosphere of H _2. The reaction is allowed to proceed at room temperature until disappearance of starting material is confirmed by TLC. The solution is filtered through a celite bed and the filtrate is concentrated in vacuo to give compounds with 4 and 6 position OH. This compound is then dissolved in pyridine (25 ml) and cooled to 0 ° C. Ph ₃ CCl (1.2 eq) is added dropwise and the reaction is allowed to proceed for 6 hours at room temperature. Ethyl acetate (50 ml) is then added and the solution is washed with 0.1 N HCl (2 × 50 ml), saturated NaHCO ₃ (1 × 50 ml) and saturated NaCl (1 × 50 ml). The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. Intermediate C is obtained by silica gel chromatography.

(Formation of 20 :)
Compound C (800 mg, 1.41 mmol) and Et ₄ NBr (353 mg, 1.69 mmol) are dissolved in DMF / CH ₂ Cl ₂ (10 ml, 1: 1 with molecular sieves) and cooled to 0 ° C. Br ₂ (298 mg, 1.86 mmol, in CH ₂ Cl ₂ ) is added dropwise at 0 ° C. to another solution of compound D (808 mg, 1.69 mmol) in CH ₂ Cl ₂ . After 30 minutes, the Br ₂ / D solution is quenched with cyclohexene (0.2 ml) and added immediately (within 10 minutes) to the C solution. The mixture is allowed to react for 65 hours at room temperature. Ethyl acetate (100 ml) is added, the solution is filtered and the filtrate is washed with saturated NaS ₂ O ₃ (2 × 50 ml) and saturated NaCl (2 × 50 ml). The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. The resulting syrup is then dissolved in ether (50 ml) and formic acid (10 ml) is added with stirring. After completion of the reaction (as confirmed by TLC), the solution is washed with saturated NaHCO ₃ (2 × 50 ml) and saturated NaCl (1 × 50 ml). The organic layer is then dried over Na ₂ SO ₄ and then evaporated to dryness. Compound 20 is then purified by silica gel chromatography.

(Formation of intermediate F :)
Compound 20 (1 g, 1.02 mmol) is dissolved in MeOH / dioxane (10 ml, 20: 1) and NaOMe (0.10 mmol) is added with stirring. The reaction is allowed to proceed for 20 hours at 50 ° C. and then 2 drops of acetic acid are added. The solution is evaporated to dryness, dissolved in ethyl ether (25 ml) and washed with saturated NaCl (1 × 50 ml). The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. The final product is purified by silica gel chromatography. This product (0.980 mmol) and Bu ₂ Sn (1.08 mmol) are suspended in MeOH (15 ml) and heated to reflux for 2 hours. The resulting clear solution is then evaporated to dryness, dissolved in pentane (10 ml) and evaporated to give a colorless foam. The foam is dissolved in 1,2-dimethoxyethane (DME, 15 ml), compound E (1.96 mmol) and CsF (1.18 mmol) are added and the reaction is stirred at room temperature for 2 hours. After 2 hours, 1M KH ₂ PO ₄ (50 ml) and KF (1 g) are added with stirring, followed by extraction with ethyl acetate (2 × 25 ml). The organic layer is washed with 10% KF (2 × 50 ml) and saturated NaCl (2 × 50 ml), dried over Na ₂ SO ₄ and evaporated to dryness under reduced pressure. Compound F is obtained via silica gel chromatography.

(Formation of 3 :)
Compound F is dissolved in CH ₃ OH (50 ml) and Pd on carbon (10%) (1/10 by weight) is added. The solution was degassed, to generate an atmosphere of H _2. The reaction is allowed to proceed at room temperature until disappearance of starting material is confirmed by TLC. The solution is filtered through a celite bed and the filtrate is concentrated in vacuo to give compound 3.

(Example 2)
(Synthesis of 21 (FIG. 4))
(Formation of intermediate H :)
G (15.0 g, 66.9 mmol) and Bu ₂ SnO (20.0 g, 80.3 mmol) are suspended in MeOH (450 ml) and heated to reflux for 2 hours. The resulting clear solution is then evaporated to dryness, dissolved in pentane and evaporated again to give a colorless foam. The foam was dissolved in 1,2-dimethoxyethane (DME, 120 ml), E (39.6 g, 100.3 mmol) and CsF (12.2 g, 80.3 mmol) were added and the reaction was allowed to come to room temperature. For 2 hours. After 2 hours, 1M KH ₂ PO ₄ (700 ml) and KF (25 g) are added with stirring, followed by extraction with ethyl acetate (3 × 250 ml). The organic layer is washed with 10% KF (2 × 250 ml) and saturated NaCl (1 × 250 ml), dried over Na ₂ SO ₄ and evaporated to dryness under reduced pressure. The compound (19.3 g, 41.2 mmol) is purified by silica gel chromatography and immediately dissolved in pyridine (210 ml) with crystalline DMAP. The solution is cooled to 0 ° C. and benzoyl chloride (52.1 g, 370.7 mmol) is added dropwise with stirring. The solution is slowly warmed to room temperature and the reaction is allowed to proceed for 20 minutes at room temperature. The solution was evaporated to dryness, dissolved in ethyl acetate (500 ml), 0.1 M HCl solution (2 x 250 ml), saturated NaHCO ₃ solution (2 x 250 ml) and saturated NaCl solution (1 x 250 ml). Wash with. The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. H is obtained by silica gel chromatography.

(Formation of Intermediate I :)
H (10.0g, 12.82mmol) and B was dissolved (6.05g, 25.64mmol) in a _CH 2 _Cl 2 (75ml), at -10 ° C., with stirring, 0.15M _CF 3 _SO 3 H (In CH ₂ Cl ₂ ) is added dropwise. The addition is stopped when the orange solution turns brown. Ethyl acetate (500 ml) is added and the solution is washed with saturated NaHCO ₃ (4 × 250 ml) and saturated NaCl (250 ml). The organic layer is then dried over Na ₂ SO ₄ and evaporated under reduced pressure. The compound (7.96 g, 9.19 mmol) is then purified by silica gel chromatography and then dissolved in DMF (55 ml). TBDMS-Cl (1.52 g, 10.1 mmol) and imidazole (0.94 g, 13.8 mmol) are then added and the reaction is allowed to proceed for 1 hour at room temperature. Ethyl acetate (250 ml) is added and the solution is washed with saturated NaHCO ₃ (5 × 250 ml) and saturated NaCl (1 × 250 ml). The organic layer is then dried over Na ₂ SO ₄ and purified by silica gel chromatography to give compound I.

(Formation of intermediate J :)
Compound I (7.71 g, 7.87 mmol) and Et ₄ NBr (2.00 g, 9.45 mmol) are dissolved in DMF / CH ₂ Cl ₂ (60 ml, containing 1: 1, 12 g molecular sieves) Cool to 0 ° C. Br ₂ (1.90 g, 11.8 mmol) in CH ₂ Cl ₂ (11 ml) is added dropwise at 0 ° C. to another solution of compound D (4.5 g, 9.45 mmol) in CH ₂ Cl _2. . After 30 minutes, the Br ₂ / D solution is quenched with cyclohexene (2.5 ml) and added immediately (within 10 minutes) to the solution of I above. The mixture is allowed to react for 65 hours at room temperature. CH ₂ Cl ₂ (250 ml) was added, the solution was filtered, and the filtrate was washed with saturated NaHCO ₃ (2 × 50 ml), 0.5M HCl (2 × 250 ml) and saturated NaCl (2 × 250 ml). ). The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. This mixture is dissolved in MeCN (85 ml) at room temperature and Et ₃ N (0.21 ml) and H ₂ SiF ₆ (1.3 ml, 35%) in MeCN (17 ml) are added and stirred for 2 hours. . CH ₂ Cl ₂ (250 ml) is added and the solution is washed with saturated NaHCO ₃ (3 × 250 ml) and saturated NaCl (1 × 250 ml). The organic layer is then dried over Na ₂ SO ₄ and then evaporated to dryness, and J is purified by silica gel chromatography.

(Formation of intermediate K :)
J (12.5 g, 9.75 mmol) is dissolved in pyridine (80 ml) and methanesulfonyl chloride (3.35 g, 29.2 mmol) is added dropwise over 5 minutes with stirring. The reaction is allowed to proceed for 30 minutes and then ethyl acetate (500 ml) is added. The solution is washed with 1N HCl (250 ml). The organic layer is dried over Na ₂ SO ₄ and evaporated. The resulting syrup (12.95 g, 9.52 mmol) is dissolved in DMF (40 ml) and NaN ₃ (4.64 g, 74.4 mmol) is added. The reaction is allowed to proceed for 35 hours at 65 ° C. under an argon atmosphere. The solution is diluted with ethyl acetate (500 ml) and washed with H ₂ O (300 ml) and saturated NaCl (150 ml). The organic layer is dried over Na ₂ SO ₃ and evaporated to dryness. The compound is purified by silica gel chromatography. The purified product (12.2 g, 9.33 mmol) was then suspended in a MeOH / H ₂ O (200 ml / 20 ml) solution and LiOH—H ₂ O (5.1 g, 121.3 mmol) was added. . The reaction is allowed to proceed for 20 hours at 65 ° C. Ethyl acetate (500 ml) was added. The solution is then washed with saturated NaCl (200 ml). The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. Compound K is purified by silica gel chromatography.

(Formation of 21 :)
Compound K (8.45 g, 9.33 mmol) is dissolved in dioxane / H ₂ O (250 ml / 50 ml) and Pd on carbon (10%) (3.4 g) is added. The solution was degassed, to generate an atmosphere of H _2. The reaction is allowed to proceed at room temperature until disappearance of starting material is confirmed by TLC. The solution is filtered through a celite bed and the filtrate is concentrated in vacuo to give compound 21.

(Example 3)
(Synthesis of 15 (FIG. 5))
(Formation of intermediate L :)
Compound 20 (10 mmol) is dissolved in CH ₂ Cl ₂ (30 ml), DMSO (20 mmol) is added and the solution is cooled to −60 ° C. Oxalyl chloride (11 mmol) is added slowly to 20 stirred solutions. The reaction is allowed to proceed for 30 minutes under N ₂ atmosphere. The solution is washed with 0.1 M HCl, saturated NaHCO ₃ and saturated NaCl. The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness under reduced pressure. The resulting syrup is taken up in tBuOH (20 ml) and 2-methyl-2-butene (10 ml) and NaH ₂ PO ₄ (20 mmol) are added with stirring. The reaction is allowed to proceed for 3 hours, then evaporated, dissolved in CH ₂ Cl ₂ and washed with 0.1 M HCl, saturated NaHCO ₃ and saturated NaCl. The resulting compound is purified by silica gel chromatography to give compound L.

(Formation of intermediate N :)
Compound L (10 mmol) is dissolved in DMF (15 ml) and compound M (10 mmol), HBTU (12 mmol) and Et ₃ N (20 mmol) are added with stirring. The reaction is allowed to proceed for 24 hours at room temperature. Ethyl acetate (100 ml) is added and the solution is washed with 0.1 M HCl (1 × 100 ml), saturated NaHCO ₃ (1 × 100 ml) and saturated NaCl (1 × 100 ml). The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. Compound N is isolated by silica gel chromatography.

(Formation of intermediate O :)
Compound N (10 mmol) is dissolved in MeOH (35 ml) and NaOMe (1 mmol) is added with stirring. The reaction is allowed to proceed for 20 hours at room temperature. The solution is evaporated to dryness, dissolved in ethyl ether (50 ml) and washed with saturated NaCl (1 × 50 ml). The organic layer is dried over Na ₂ SO ₄ and evaporated to dryness. The final product is purified by silica gel chromatography. The product (0.980 mmol) and Bu ₂ Sn (1.08 mmol) are suspended in MeOH (15 ml) and heated to reflux for 2 hours. The resulting clear solution is then evaporated to dryness, dissolved in pentane (10 ml) and evaporated to give a colorless foam. The foam is dissolved in 1,2-dimethoxyethane (DME, 15 ml), compound E (1.96 mmol) and CsF (1.18 mmol) are added and the reaction is stirred at room temperature for 2 hours. After 2 hours, 1M KH ₂ PO ₄ (50 ml) and KF (1 g) are added with stirring, followed by extraction with ethyl acetate (2 × 25 ml). The organic layer is washed with 10% KF (2 × 50 ml) and saturated NaCl (2 × 50 ml), dried over Na ₂ SO ₄ and evaporated to dryness under reduced pressure. Compound O is obtained by silica gel chromatography.

(Formation of 15 :)
Compound O (9 mmol) is dissolved in MeOH (200 ml) and Pd on carbon (10%) (3 g) is added. The solution was degassed, to generate an atmosphere of H _2. The reaction is allowed to proceed at room temperature until disappearance of starting material is confirmed by TLC. The solution is filtered through a celite bed and the filtrate is concentrated in vacuo to give compound 15.

Example 4
(Acylation of 21 (FIG. 6))
(Reaction with 21 acid chlorides :)
Compound 21 (20 mg, 0.033 mmol) is dissolved in a solution of THF / H ₂ O (2 ml, 1: 1) containing 1N NaOH (pH adjusted between 8-10) and cooled to 0 ° C. To do. Cyclohexylcarbonyl chloride (0.049 mmol) is then added dropwise with stirring. The reaction is continued at 0 ° C. for 3 hours. The solution is quenched with ice and the solution is evaporated to dryness. Compound 1 is purified by reverse phase chromatography.

(Reaction with 21 isocyanate :)
Compound 21 (30 mg, 0.049 mmol) is dissolved in 0.5N aqueous NaOH (1 ml) and cooled to 0 ° C. Ethyl isocyanate (1.2 eq) is then added dropwise with stirring. The reaction is continued for 3 hours at room temperature. The solution is quenched with ice and the solution is evaporated to dryness. Compound 2 is purified by reverse phase chromatography.

(Reaction of 21 with chloroorthoformate :)
Compound 21 (20 mg, 0.033 mmol) is dissolved in a THF / H ₂ O (2 ml, 1: 1) solution containing NaOH (pH adjusted between 8-10) and cooled to 0 ° C. . Benzylchloroorthoformate (0.049 mmol) is then added dropwise with stirring. The reaction is continued at 0 ° C. for 3 hours. The solution is quenched with ice and the solution is evaporated to dryness. Compound 11 is purified by reverse phase chromatography.

(Reaction of 21 with sulfonyl chloride :)
Compound 21 (20 mg, 0.033 mmol) is dissolved in saturated aqueous NaHCO ₃ / toluene (2 ml, 1: 1) and cooled to 0 ° C. P-Toluenesulfonyl chloride (0.049 mmol) is then added dropwise with stirring. The reaction is continued at 0 ° C. for 3 hours. The solution is quenched with ice and the solution is evaporated to dryness. Compound 9 is purified by reverse phase chromatography.

(Example 5)
(Synthesis of Compound 22 (FIG. 7))
Compound 1 (2.77 mmol) is dissolved in a solution of compound 2 (5.55 mmol) in diethyl ether / dichloromethane (1 ml, 1: 1 mixture). 0.1M TMSOTf (2.8 ml) solution is then added with stirring. After 30 minutes, the solution is neutralized by the addition of sodium bicarbonate (0.5 g), filtered and concentrated to dryness. Compound 3 is purified by column chromatography (toluene / acetone, 4: 1 mixture).

Compound 3 (1.96 mmol) is stirred with 4A molecular sieves, TBABr (968 mmol) and CuBr ₂ (529 mmol) in DMF / dichloromethane (50 ml, 1: 5 mixture) for 1 hour. Compound 4 (2.94 mmol) is dissolved in dichloromethane (5 ml) and added dropwise to the above solution. The solution is allowed to react for 24 hours, filtered and washed with saturated sodium bicarbonate and water (50 ml each). The organic layer is isolated, dried over sodium sulfate and evaporated to dryness. Compound 5 is purified by column chromatography (hexane / ethyl acetate, 4: 1 mixture).

  Compound 22 is obtained by dissolving compound 5 in methanol followed by addition of 1M sodium methoxide solution. The solution is allowed to react for 3-5 hours and then neutralized with Amberlite IR-120 resin, filtered and concentrated in vacuo. Compound 22 is purified by column chromatography (dichloromethane / methanol, 20: 1 mixture).

(Example 6)
(Synthesis of Compounds 4, 6, 7, 8, 10, 12, 13, 14, 16, 17, 18, 19 and 20)
Starting from compound 21 (Example 2; FIG. 4), these compounds were obtained from the literature (Helvetica Chimica Acta Vol. 83, pp. 2893-2907, 2000; Angew Chem. Int. Ed. Vol. 40, No. 19). , Pp. 3644-3647, 2001).

(Example 7)
(Synthesis of Compounds 23, 24, 25, 26, 27 and 28)
Synthesis of intermediate 23: Compound i (2.77 mmol) is dissolved in a solution of diethyl ether / dichloromethane (1 ml, 1: 1 mixture) containing compound 2 (5.55 mmol). A 0.1 M TMSOTf (2.8 ml) solution is then added dropwise with stirring. After 30 minutes, the solution is neutralized by the addition of sodium bicarbonate (0.5 g), filtered and concentrated to dryness. 3 is purified by silica gel chromatography to give compound iii.

Compound iii (1.96 mmol) is stirred with 4A molecular sieves, TBABr (968 mmol) and CuBr ₂ (529 mmol) in DMF / dichloromethane (50 ml, 1: 5 mixture) for 1 hour. Compound xv (2.94 mmol) is dissolved in dichloromethane (5 ml) and added dropwise to the above solution. The solution is allowed to react for 24 hours, filtered and washed with saturated sodium bicarbonate and water (50 ml each). The organic layer is isolated, dried over sodium sulfate and evaporated to dryness. iv is then purified by silica gel chromatography.

  Compound iv is de-O-acetylated with 0.1 N NaOMe in MeOH to give v and then treated with dibutyltin oxide in MeOH under reflux conditions to give vi (after evaporation of the solvent), The vi is used in the next step without further purification.

  Compound vi (1 mmol) is dissolved in DME (20 ml) and then compound vii (2.5 mmol) and CsF (1.4 mmol) are added. The resulting reaction mixture is stirred at room temperature for 8 hours, diluted with ethyl acetate and the organic layer is washed with water. The organic layer is concentrated to dryness and purified by silica gel chromatography to give compound viii.

  Synthesis of compound 23: Compound viii is hydrogenated in the presence of palladium in carbon to give compound 23.

  Synthesis of intermediate xx: Starting from N-acetylglucosamine (5,50 g), compound xx (50% overall process yield) was prepared according to published procedures (Bioorg. Med. Chem. Lett. 11, pp. 199 923-925, 2001; Carbohydr. Res. 197, 75, 1990).

  Synthesis of intermediate xv: Compound xv (15 g) is synthesized from L-fucose according to the procedure described in the literature (Carbohydr. Res. 201, 15-30, 1990).

  Synthesis of intermediate xix: Compound xix is prepared from commercially available β-D-galactose pentaacetate as described (WO9701569; Chem. Astr., 126 186312, 1997).

Synthesis of intermediate xxxiv: commercially available N-acetylneuraminic acid (xxx, 10 g) is suspended in MeOH—H ₂ O (60 ml; 9: 1) and an aqueous solution of cesium carbonate is added To adjust its pH to 8.1. The solvent is removed and the residue is repeatedly evaporated with ethanol and then with hexane. The material is dissolved in DMF (65 ml) and benzyl bromide (3.5 ml) is added within 20 minutes. The mixture is stirred for 16 hours, after which dichloromethane (100 ml) is added and washed with water (50 ml). The solvent is distilled off and purified by silica gel chromatography to give xxx in 68% yield.

  To a solution of compound xxxi (7 g) in pyridine (50 ml) is added acetic anhydride (48 ml) and the reaction mixture is stirred at room temperature for 16 hours. The solvent is distilled off and the residue (xxxii) is dissolved in dry DMF (25 ml). To the mixture is added ground ammonium carbonate (2 g) and the mixture is stirred at 28 degrees Celsius for 12 hours. This mixture is added to an ice-cold solution of 1N HCl in water (50 ml) and dichloromethane (100 ml) is added. After solvent extraction, the organic layer is distilled off and then dried under vacuum for 24 hours. The residue is purified by silica gel chromatography to give xxxiii in 71% yield.

  Compound xxxiii is dissolved in dry dichloromethane and 2,6-di-tert-butylpyridine (5 g) is added. The solution is cooled to −20 degrees Celsius and trifluoromethanesulfonic anhydride (7 g) is added in portions over 10 minutes. The mixture is stirred for 4 hours, diluted with dichloromethane (100 ml) and added to a solution of potassium hydrogen phosphate (500 ml). The layers are separated and the organic layer is dried (sodium sulfate) and the solvent is evaporated to give xxxiv, which is used in the next step without further purification.

  Synthesis of intermediate xxxv: To a mixture of compound xx (10 g) and compound xv (15 g) in dichloromethane (100 ml) is added molecular sieves (4A, 8 g). Stir at room temperature for 1 hour, then add tetraethylammonium bromide (5 g). A solution of bromine (1 g) in dichloromethane (25 ml) is added dropwise during 1 hour. The reaction mixture is kept stirring for 3 hours, filtered through a celite bed and washed successively with cold water, a saturated aqueous solution of sodium bicarbonate and water. The solvent is distilled off and subjected to silica gel chromatography.

  The product was treated with sodium cyanoborohydride in THF and HCl in ether to give compound xxxv in 70% overall process yield after silica gel chromatography.

  Synthesis of intermediate xxxvi: To a mixture of xxxv (10 g) and xix (7 g) in dichloromethane (80 ml) is added N-iodosuccinimide (15 g) and molecular sieves (4A, 8 g). The reaction mixture is placed in an ice bath. The solution is stirred at 0-5 degrees for 30 minutes and a solution of trifluoromethanesulfonic acid (0.2 ml) in dichloromethane (25 ml) is added dropwise with stirring for 1 hour. Stirring is continued for 2 hours, filtered through a celite bed and washed sequentially with cold water, a cold saturated solution of sodium bicarbonate and water. Evaporate the solvent and purify by silica gel chromatography to give compound xxxvi in 68% yield.

  Synthesis of intermediate xxxvii: Compound xxxvi (8 g) is treated with 0.05 N NaOEt in MeOH (100 ml) for 4 hours and after neutralization with IR120 (hydrogen type) resin, the reaction mixture is filtered off. The solvent is distilled off to give compound xxxvii in 96% yield.

  Synthesis of intermediate xxxviii: Compound xxxvii (5 g) is treated with dibutyltin oxide (1 g) in MeOH for 4 h under reflux. The solvent is distilled off and co-evaporated several times with toluene and finally the residue is dried under high vacuum for 24 hours.

  The crude reaction mixture is dissolved in dimethoxyethane (DME, 100 ml) and CsF (1.7 g) and compound xxxiv (2.5 g) are added. The reaction mixture is stirred at room temperature for 8 hours and ethyl acetate (100 ml) is added. The organic layer is washed with water and the organic solvent is distilled off. The product is purified by silica gel chromatography to give xxxviii in 64% yield.

  Synthesis of intermediate xxxix: Compound xxxviii (2 g) was de-O-acetylated using 0.01 N NaOMe in MeOH (100 ml, 1 h) and the crude reaction mixture was neutralized with IR120 (hydrogen type) resin. The solvent is distilled off.

  The product from the above reaction is dissolved in dioxane-water (1: 1, 50 ml) and 10% PD-C is added. The reaction mixture is stirred vigorously under a hydrogen atmosphere for 22 hours, filtered through a celite bed and the solvent is distilled off. Silica gel chromatography of the resulting syrup gives xxxix in 77% yield.

Synthesis of Compound 24: To a solution of the compound xxxix in _{MeOH-H 2} O, was added a solution of NaOMe in MeOH, and the reaction mixture is stirred for 2 hours at room temperature. Neutralization with IR120 resin and evaporation to dryness gives compound 24.

  Synthesis of xxx: Compound xxxix is treated with ethylenediamine (500 mg) at 70 degrees Celsius for 4 hours. The solvent is distilled off and the syrupy residue is purified by silica gel chromatography to give compound xxx in 77% yield.

  Synthesis of compound xxxv: 3-nitrobenzyl iodide is added to a commercially available aqueous solution of 8-aminonaphthalene-1,3,5-trisulfonic acid (xxxxi) (pH 11) at room temperature with stirring. The pH of the solution is adjusted to 1 and the product xxxiii is precipitated from ethanol after evaporation of the solvent.

  Platinum-catalyzed hydrogenation of compound xxxiii gives compound xxxiv in 96% yield.

  To a solution of compound xxxiv in phosphate buffer (pH 7.1) is added commercially available squaric acid and the reaction mixture is stirred at room temperature for 4 hours. It is then purified by reverse phase HPLC to give xxxv.

  Synthesis of compound 25: Compound xxx (0.2 g) is dissolved in carbonate buffer (2 ml, pH 8.8) and compound xxxv (0.4 g) is added. The reaction mixture is stirred at room temperature for 24 hours. Another batch (0.2 g) of compound xxxv is added and stirring is continued for 20 hours at room temperature. The solvent is distilled off and the mixture is purified by reverse phase HPLC to give 25.

  Synthesis of compound 26: Compound xxxix (0.1 g) is reacted with compound xxxiv to give compound 26 after purification by HPLC.

Synthesis of intermediate xxxxiii: commercially available compound xxxvi (4- (4,4,5,5-tetramethyl- [1,3,2] dioxaborolan-2-yl) benzoic acid, 1 equivalent) and KOAc (3 eq) is taken up in THF (25 ml). To the resulting slurry is added PdCl ₂ and commercially available p-bromonitrobenzene (xxxvii, 1.2 eq) with stirring, and the mixture is heated gently to 80 degrees Celsius. After 6 hours, the reaction mixture is evaporated to dryness, dissolved in dichloromethane (30 ml) and washed with distilled water and a saturated solution of sodium bicarbonate. The resulting biphenyl compound xxxviii is directly subjected to the next step.

  Compound xxxviii (1 eq), dimethylaminopyridine (catalytic amount, 1 crystal) and EDCl (1.05 eq) are dissolved in DMF or THF (20 ml) and allowed to react at room temperature for 10 minutes. The commercially available compound xxxix (8-aminonaphthalene-1,3,5-trisulfonic acid) is added to the reaction mixture with stirring and the reaction is allowed to proceed for 48 hours at room temperature under nitrogen. . The reaction mixture is then evaporated to dryness and purified by reverse phase chromatography to give compound xxxxxx.

  To a solution of compound xxxxxxx in EtOAc, PD-C is added and the reaction mixture is stirred for 2 hours under a hydrogen atmosphere. The reaction mixture is filtered through a celite bed and evaporated to dryness to give compound xxxxxxx.

  To a solution of compound xxxxxxx in phosphate buffer (pH 7.1) is added commercially available squaric acid and the reaction mixture is stirred at room temperature for 4 hours. It is then purified by reverse phase HPLC to give xxxiii.

  Synthesis of Compound 27: Compound xxx (0.2 g) is dissolved in carbonate buffer (2 ml, pH 8.8) and Compound xxxii (0.4 g) is added. The reaction mixture is stirred at room temperature for 24 hours. Another batch (0.2 g) of compound xxxiii is added and stirring is continued for 20 hours at room temperature. The solvent is distilled off and the mixture is purified by reverse phase HPLC to give 27.

  Synthesis of compound 28: Compound xxxix is reacted with compound xxxxxxx to give compound 28, which is purified by HPLC.

(References on synthesis)
Synthetic protocols for the preparation of specific compounds included within this application are exemplified in the following references: Helvetica Chema Acta Vol. 83, pp. 2893-2907 (2000) and Angew. Chem. Int. Ed. Vol. 40, no. 19, pp. 3644-3647 (2001).

(Example 8)
(Assay)
Purified oligosaccharide (new glycoprotein) chemically conjugated to albumin is coated into plastic microtiter wells. After blocking with BSA, the wells are then incubated with purified PA-IIL lectin and allowed to bind to purified PA-IIL lectin. Bound PA-IIL lectin is detected with anti-PA-IIL rabbit antiserum followed by HRP-labeled anti-rabbit Ig and TMB reagent using color development with 1M H ₃ PO ₄ (FIG. 18). Structure and activity are presented in Table 1.

Table 1. Structure of immobilized neutral oligosaccharide screened for binding to PA-IIL lectin
The results of screening for immobilized neutral oligosaccharide are shown in FIG. 18 and summarized below. Type 1 lacto-series chains (eg, Le ^a structures) bind to PA-IIL lectins better than lacto-series type 2 chains (eg, Le ^x structures). As in Le ^a / Le ^x , extension of the type 1 structure results in the most active compounds in this series.

（表２．ＰＡ−ＩＩＬレクチンへの結合についてスクリーニングされた固定化酸性オリゴ糖の構造）
固定化酸性オリゴ糖のスクリーニングの結果を、図１９に記載し、以下にまとめる。フコースを含むシアリル化オリゴ糖のみが、ＰＡ−ＩＩＬレクチンに結合する。ジシアリルＬｅ^ａ上の第２のシアリル基は、結合を阻害するように見える。

Table 2. Structure of immobilized acidic oligosaccharide screened for binding to PA-IIL lectin
The results of screening for immobilized acidic oligosaccharides are described in FIG. 19 and summarized below. Only sialylated oligosaccharides containing fucose bind to PA-IIL lectins. Disialyl second sialyl groups on Le ^a appears to inhibit the binding.

アルブミンに化学的に結合させた精製オリゴ糖（新しい糖タンパク質）を、プラスチック製マイクロタイターウェル中にコーティングする。ＢＳＡでブロックした後、そのウェルを次いで精製ＰＡ−ＩＩＬレクチンとともにインキュベートし、精製ＰＡ−ＩＩＬレクチンに結合させる。結合されたＰＡ−ＩＩＬレクチンを、１Ｍ Ｈ_３ＰＯ_４による発色を用いて、抗ＰＡ−ＩＩＬウサギ抗血清、続いてＨＲＰ標識化抗ウサギＩｇおよびＴＭＢ試薬で検出する（図１９）。構造および活性を表２に提示する。

Purified oligosaccharide (new glycoprotein) chemically conjugated to albumin is coated into plastic microtiter wells. After blocking with BSA, the wells are then incubated with purified PA-IIL lectin and allowed to bind to purified PA-IIL lectin. Bound PA-IIL lectin is detected with anti-PA-IIL rabbit antiserum followed by HRP-labeled anti-rabbit Ig and TMB reagents using color development with 1M H ₃ PO ₄ (FIG. 19). Structure and activity are presented in Table 2.

Fucose biotinylated polymer (Fuc-PAA-biotin, GlycoTech Corp.) is coupled with HRP labeled streptavidin (KPL labs) at 4 ° C. overnight. PA-IIL lectin is immobilized in plastic microtiter wells and reacted with HRP labeled fucosylated polymer. Binding is detected with TMB substrate followed by color development with 1M H ₃ PO ₄ (FIG. 20). The conditions chosen for the assay were incubation of the inhibitor with a coating of 3 μg PA-IIL per ml and 2 μg fucosylated polymer per ml.

Compound 23 inhibited PA-IIL with an IC ₅₀ of 420 nM (FIG. 22), whereas the natural saccharide inhibitor mannose inhibited PA-IIL lectin with an IC ₅₀ of 95 μM and was a negative control saccharide. Some galactose showed no inhibition (data not shown).

  While specific embodiments of the present invention have been described herein for purposes of illustration, it will be appreciated that various modifications can be made without departing from the spirit and scope of the invention. It is understood from.

FIG. 1A shows the structure of a sugar mimetic compound. FIG. 1B shows the structure of the sugar mimetic compound. FIG. 1C shows the structure of the sugar mimetic compound. FIG. 1D shows the structure of a sugar mimetic compound. FIG. 1E shows the structure of a sugar mimetic compound. FIG. 2A shows the structure of additional glycomimetic compounds. FIG. 2B shows the structure of additional glycomimetic compounds. FIG. 3 illustrates the synthesis of a representative glycomimetic compound 3. FIG. 4 illustrates the synthesis of sugar mimetic compound 21. FIG. 5 illustrates the synthesis of a representative sugar mimetic compound 15. FIG. 6 illustrates the acylation of intermediate 21 to give various representative sugar mimetic compounds. FIG. 7 illustrates the synthesis of compound 22. FIG. 8A illustrates the synthesis of compounds 4, 6, 7, 8, 10, and 12. FIG. 8B illustrates the synthesis of compounds 13, 14, 16, 17, 18, and 19. FIG. 9 illustrates the synthesis of compounds 22 and 23. FIG. 10 illustrates the synthesis of the intermediate. FIG. 11 illustrates the synthesis of intermediate and compound 24. FIG. 12 illustrates the synthesis of intermediates xxxiv and xxxv. FIG. 13 illustrates the synthesis of compound 25. FIG. 14 illustrates the synthesis of compound 26. FIG. 15 illustrates the synthesis of intermediates xxxxxxxx and xxxiiii. FIG. 16 illustrates the synthesis of compound 27. FIG. 17 illustrates the synthesis of compound 28. FIG. 18 illustrates the binding of PA-IIL lectin to the immobilized neutral sugar chain structure. FIG. 19 illustrates the binding of PA-IIL lectin to the immobilized acidic sugar chain structure. FIG. 20 illustrates determination of assay conditions for IC ₅₀ values for PA-IIL lectin inhibition. FIG. 21 shows a schematic representation of an assay developed to determine IC ₅₀ values for PA-IIL lectin glycomimetic inhibitors. FIG. 22 illustrates inhibition of PA-IIL lectin by glycomimetic compound 23.

Claims (17)

A method of inhibiting a Pseudomonas bacterium in a warm-blooded animal comprising the step of administering to the animal a compound comprising the compound described in FIG. 1 or FIG. 2 in an amount effective to inhibit the bacterium. Method. The method of claim 1, wherein the compound is conjugated to a therapeutic agent. 3. The method of claim 1 or 2, wherein the compound is combined with a pharmaceutically acceptable carrier or diluent. The method according to claim 1, wherein the bacterium is Pseudomonas aeruginosa. A conjugate comprising a therapeutic agent conjugated to the compound of FIG. 1 or FIG. 6. The conjugate according to claim 5, wherein the conjugate is combined with a pharmaceutically acceptable carrier or diluent. A method of detecting a Pseudomonas bacterium comprising: a diagnostic agent conjugated to a compound comprising a compound described in FIG. 1 or FIG. 2 and a sample, wherein the compound is Contacting under conditions sufficient for binding; and detecting the agent present in the sample, wherein the presence of the agent in the sample indicates the presence of Pseudomonas bacteria A method comprising the steps. The method according to claim 7, wherein the bacterium is Pseudomonas aeruginosa. A method of immobilizing a Pseudomonas bacterium on a solid support, the method being described in FIG. 1 or FIG. 2 immobilized on a solid support and a Pseudomonas bacterium-containing sample under conditions sufficient for binding. Contacting with a compound comprising a compound of: and separating the sample from the solid support. A compound comprising the compound of FIG. 1 or FIG. 2 for use in a method of inhibiting Pseudomonas bacteria. 11. A compound according to claim 10 in combination with a pharmaceutically acceptable carrier or diluent. The compound according to claim 10 or 11, wherein the bacterium is Pseudomonas aeruginosa. 7. A conjugate according to claim 5 or 6 for use in a method of inhibiting Pseudomonas bacteria. 14. The conjugate according to claim 13, wherein the bacterium is Pseudomonas aeruginosa. Use of a compound comprising the compound described in FIG. 1 or FIG. 2 in the preparation of a medicament for the inhibition of Pseudomonas bacteria. Use of a conjugate according to claim 5 in the preparation of a medicament for the inhibition of Pseudomonas bacteria. Use according to claim 15 or 16, wherein the bacterium is Pseudomonas aeruginosa.

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