WO2000035904A1 - INHIBITEURS DE β-LACTAMASES ET LEUR UTILISATION - Google Patents

INHIBITEURS DE β-LACTAMASES ET LEUR UTILISATION Download PDF

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Publication number
WO2000035904A1
WO2000035904A1 PCT/US1999/029958 US9929958W WO0035904A1 WO 2000035904 A1 WO2000035904 A1 WO 2000035904A1 US 9929958 W US9929958 W US 9929958W WO 0035904 A1 WO0035904 A1 WO 0035904A1
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Prior art keywords
lower alkyl
halogen atoms
substituted
compound
alkene
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PCT/US1999/029958
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English (en)
Inventor
Brian K. Shoichet
Maria Paola Costi
Donatella Tondi
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Northwestern University
Università Degli Studi Di Modena E Reggio Emilia
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Priority to AU31243/00A priority Critical patent/AU3124300A/en
Publication of WO2000035904A1 publication Critical patent/WO2000035904A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/69—Boron compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
    • C07F5/02—Boron compounds
    • C07F5/025—Boronic and borinic acid compounds

Definitions

  • ⁇ -lactam drugs e.g. , amoxicillin, cephalothin, clavulanate, aztreonam
  • ⁇ -lactams target enzymes that are unique to bacteria and are thus highly selective. They have been widely prescribed.
  • ⁇ -lactams are the first choice for treatment in 45 of 78 common bacterial infections (Goodman & Gilman's The Pharmacological Basis of Therapeutics (Hardman et al., eds., McGraw-Hill, New York, 1996)).
  • the evolution of resistance to these drugs has raised the cost of antibiotic therapy and reduced its effectiveness, leading to increased rates of morbidity and mortality.
  • ⁇ -lactam antibiotics inhibit bacterial cell wall biosynthesis (Tomasz, Rev. Infect.
  • PBPs penicillin binding proteins
  • TEM-1 and AmpC are two ⁇ -lactamases from Escherichia coli.
  • E. coli is an important pathogen in its own right. It is the most common cause of gram-negative bacterial infection in humans (Levine, New Engl. J. Med, 313, 445-447 (1985)), and is the most prevalent hospital-acquired infection (Thornsberry, Pharmacotherapy, 15, S3-8 ( 1995)).
  • E. coli that carry TEM- 1 , or for which AmpC production has been derepressed, are resistant to ⁇ -lactam treatment. As of 1992, as many of 30% of community-isolated E. coli and 40-50% of hospital-acquired E.
  • T ⁇ M-1 and AmpC are major forms of plasmid-based and chromosomal ⁇ -lactamases and are responsible for resistance in a broad host range.
  • the versions of TEM and AmpC (Galleni, et al., Biochem. J, 250, 753-760 (1988)) in other bacterial species share high sequence identity to TEM-1 and AmpC from E. coli.
  • TEM-1 structurally and catalytically resembles the class A ⁇ -lactamase from Staphlococcus aureus.
  • the structures of AmpC from Citrobacter freundii and Enterobacter cloacae have been determined, and they closely resemble the structure of the E. coli enzyme (Usher et al., Biochemistry, 37, 16082-16092 (1998)).
  • soil bacteria have produced ⁇ -lactams that resist hydrolysis by ⁇ -lactamases or have produced ⁇ -lactams that inhibit the ⁇ -lactamases.
  • Streptomyces clavuligeris makes several ⁇ -lactams, including clavulanic acid, a clinically used inhibitor of class A ⁇ -lactamases such as TEM-1.
  • Chromobacterium violaceum makes aztreonam, a clinically used monobactam that resists hydrolysis by many ⁇ -lactamases.
  • bacteria have been able to respond rapidly with "new" resistance mechanisms to ⁇ -lactams, and indeed many classes of antibiotics, is that the mechanisms are not in fact new.
  • Novel inhibitors would escape detection by ⁇ -lactam sensor proteins that up-regulate ⁇ -lactamase transcription, and may be unaffected by porin mutations that limit the access of ⁇ -lactams to PBPs. Such inhibitors would allow current ⁇ -lactam drugs to work against bacteria where ⁇ -lactamases provide the dominant resistance mechanism.
  • boric acid and certain phenyl boronic acids are inhibitors of certain ⁇ -lactamases. See, Kiener and Waley, Biochem. J, 169, 197-204 (1978) (boric acid, phenylboronic acid (2FDB) and w-aminophenylboronate (MAPB));
  • the invention provides non- ⁇ -lactam inhibitors of ⁇ -lactamases.
  • the invention provides ⁇ -lactamase inhibitors having the formula:
  • t is N-lower alkyl, a cyclic alkene or a heterocyclic alkene, wherein the cyclic alkene and heterocyclic alkene may be substituted with one or more substituents R 2 ; and each R 2 is independently H, a halogen atom, lower, alkyl, lower alkyl substituted with one or more halogen atoms, NH 2 , NO, NO 2 , N-lower alkyl, N-lower alkyl substituted with one or more halogen atoms, OH, O-lower alkyl, O-lower alkyl substituted with one more halogen atoms, CO-lower alkyl, CO-lower alkyl substituted with one or more halogen atoms, COOH, lower alkyl-COOH, COO-lower alkyl, CONH 2 , CON-lower alkyl, SO 3 H, SO 2 NH 2 , SO 2 N-lower alkyl, or B
  • the invention also provides a method of treating a ⁇ -lactam-antibiotic-resistant bacterial infection.
  • the method comprises administering to an animal suffering from such an infection an effective amount of a ⁇ -lactamase inhibitor of formula (1), or a pharmaceutically-acceptable salt thereof, and an effective amount of a ⁇ -lactam antibiotic.
  • compositions comprising compounds of formula (1), or pharmaceutically-acceptable salts thereof, and a pharmaceutically-acceptable carrier.
  • the pharmaceutical compositions may also comprise ⁇ -lactam antibiotics.
  • FIGS. 1A-B Diagrams of the synthesis of compounds of formula (1).
  • Fiaure 2 Diagram of scheme for the synthesis of compounds 15-26.
  • “Lower alkyl” means a straight-chain or branched-chain alkyl containing 1-4 carbon atoms.
  • N-lower alkyl means N with one or more lower alkyls attached, such as -NHCH 3 and -N(CH 3 ) 2 .
  • O-lower alkyl means O with a lower alkyl attached, such as -OCH,.
  • Lower alkyl-COOH means -COOH preceded by a lower alkyl radical, such as -CH 2 COOH.
  • COO-lower alkyl means -COO- having a lower alkyl attached to the O, such as p — C-0-CH 3
  • CON-lower alkyl means CON having one or more lower alkyls attached to the N, such as o
  • “SO 2 N-lower alkyl” means SO 2 N having one or more lower alkyls attached to the N, such as -SO 2 NHCH 3 .
  • “Cyclic alkene” means a structure containing 1 or 2 rings, each ring containing 5 or 6 carbon atoms and at least one double bond. One or both of the rings may be aromatic. If the cyclic alkene contains more than one ring, the rings may be fused, connected by a bond, or connected by a linker, L. Preferably, L is a short chain containing up to six atoms in the chain.
  • CH CH-), lower alkyne (e.g., -C ⁇ C-), and combinations thereof.
  • Heterocyclic alkene means a cyclic alkene, as defined above, wherein one or both of the rings contain(s) one or more S, N or O atoms.
  • “Lower alkene” means a straight-chain or branched-chain alkene containing 2-4 carbon atoms.
  • “Lower alkyne” means a straight-chain or branched-chain alkyne containing 2-4 carbon atoms.
  • Compounds of formula (1) according to the invention include all optical isomers.
  • Preferred compounds of formula (1) are those wherein R,. is an cyclic alkene or heterocyclic alkene containing 2 rings linked by a linker L.
  • the preferred linkers are
  • the compounds of formula (1) can be synthesized as described below. Unless otherwise noted, the various chemicals used in the syntheses described below are available from commercial sources including Aldrich Chemical, Milwaukee, WI, Lancaster
  • the synthesis depicted in Figure 1A is performed as follows.
  • the polystyrene resin, P functionalized with a 3-benzyloxy-l,2-propanediol (5 mmol; 1 mmol/g; 5g) as described in Leznoff and Wong, Can. J. Chem., 51:3756-3764 (1973), is swollen in anhydrous acetone.
  • functionalized resins can be purchased from Novabiochem.
  • 3-aminophenylboronic acid (10 mmol; 0.93g) is dissolved in anhydrous acetone and then added to the resin suspension, and the suspension is agitated.
  • Anhydrous sodium sulfate can be used to absorb liberated water.
  • the resin is filtered and washed several times with water (to remove sodium sulfate and ethanol) and then with ether.
  • the resin is divided into batches (20 mg of resin each).
  • the resin is suspended in N-methyl pyrrolidone (NMP) and then the sulfonyl chloride (10 mmol), previously dissolved in NMP, is added to the suspension.
  • Diisopropylamine (DLEA; 20 mmol) is added.
  • the reactions are mixed for 3-4 hours.
  • excess reagents are washed away.
  • the final products are cleaved from the resin using acidic conditions and the resin is then separated by filtration.
  • the solution is evaporated or extracted to give the final product.
  • the compounds of formula ( 1 ) may contain an acidic or basic functional group and are, thus, capable of forming pharmaceutically-acceptable salts with pharmaceutically- acceptable acids and bases.
  • pharmaceutically-acceptable salts in these instances refers to the relatively non-toxic, inorganic and organic acid and base addition salts of compounds of formula (1). These salts can be prepared by reacting the purified compound with a suitable acid or base.
  • Suitable bases include the hydroxide, carbonate or bicarbonate of a pharmaceutically-acceptable metal cation, ammonia, or a pharmaceutically-acceptable organic primary, secondary or tertiary amine.
  • Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like.
  • Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.
  • Representative acid addition salts include the hydrobromide, hydrochloride, sulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthalate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like.
  • the compounds of formula ( 1 ), and the pharmaceutically-acceptable salts thereof, are inhibitors of ⁇ -lactamases.
  • the compounds of formula (1) may also prevent transcriptional up-regulation of ⁇ -lactamases and may be antibacterial by themselves, since it is likely that they will bind to PBPs which resemble ⁇ -lactamases.
  • Assays for the inhibition of ⁇ -lactamase activity are well known in the art. For instance, the ability of a compound to inhibit ⁇ -lactamase activity in a standard enzyme inhibition assay may be used (see, e.g. , Example 2 below and M. G. Page, Biochem J. 295 (Pt. 1) 295-304 (1993)).
  • ⁇ -lactamases for use in such assays may be purified from bacterial sources or, preferably, are produced by recombinant DNA techniques, since genes and cDNA clones coding for many ⁇ -lactamases are known. See, e.g., S.J.
  • a porin mutation reduces the ability of ⁇ -lactam antibiotics to enter bacterial cells in which the mutations occur, thereby making the bacteria resistant to these antibiotics.
  • the presence of a porin mutation can be detected by polymerase chain reaction analysis of porin genes, polyacrylamide gel electrophoresis of a preparation obtained by mild osmotic shock (e.g. , treatment with hypotonic solution containing EDTA, followed by gentle centrifugation and separation of the supernatant) of the bacteria (absence of a protein of the appropriate molecular weight being indicative of a porin mutation), or by determining resistance to infection by bacteriophage Tul A (a standard test for OmpF " porin mutations).
  • ⁇ -lactam-antibiotic-resistant bacterial infections can be used herein to refer to an infection caused by bacteria resistant to treatment with ⁇ -lactam antibiotics due primarily to the action of a ⁇ - lactamase.
  • Resistance to ⁇ -lactam antibiotics can be determined by standard antibiotic sensitivity testing.
  • the presence of ⁇ -lactamase activity can be determined as is well known in the art (see above).
  • the sensitivity of a particular bacterium to the combination of a compound of formula (1), or a pharmaceutically- acceptable salt thereof, and a ⁇ -lactam antibiotic can be determined by standard antibiotic sensitivity testing methods.
  • an animal suffering from such an infection is given an effective amount of a compound of formula (1), or a pharmaceutically-acceptable salt thereof, and an effective amount of a ⁇ -lactam antibiotic.
  • the compound of formula (1), or a pharmaceutically-acceptable salt thereof, and the antibiotic may be give- wately or together. When administered together, they may be contained in separate pharmaceutical compositions or may be in the same pharmaceutical composition.
  • ⁇ -lactam antibiotics include cephalosporins (e.g. , cephalothin), penicillins (e.g., amoxicillin), monobactams (e.g. , aztreonam), carbapenems (e.g. , imipenem), carbacephems (loracarbef), and others, ⁇ -lactam antibiotics are effective.
  • cephalosporins e.g. , cephalothin
  • penicillins e.g., amoxicillin
  • monobactams e.g. , aztreonam
  • carbapenems e.g. , imipenem
  • carbacephems loracarbef
  • ⁇ -lactam antibiotics are effective
  • bacteria of the genus Staphylococcus such as Staphylococcus aureus and Staphylococcus epidermis
  • Streptococcus such as Streptococcus agalactine, Streptococcus penumoniae and Streptococcus faecalis
  • Micrococcus such $ Micrococcus luteus
  • Bacillus such as Bacillus subtilis
  • Listerella such as Listerella monocytogenes
  • Escherichia such as Escherichia coli
  • Klebsiella such as Klebsiella pneumoniae
  • Proteus such as Proteus mirabilis and Proteus vulgaris
  • Salmonella such as Salmonella typhosa
  • Shigella such as Shigella sonnet
  • Enterobacter such as Enterobacter aerogenes and Enterobacter facia
  • Effective doses and modes of administration of ⁇ -lactam antibiotics are known in the art or may be determined empirically as described below for the compounds of formula (1).
  • an effective amount of a compound of formula (1), or a pharmaceutically-acceptable salt thereof is administered to the animal, in combination with a ⁇ -lactam antibiotic.
  • Effective dosage forms, modes of administration and dosage amounts of a compound of formula (1) may be determined empirically, and making such determinations is within the skill of the art.
  • the dosage amount will vary with the activity of the particular compound employed, the severity of the bacterial infection, the route of administration, the rate of excretion of the compound, the duration of the treatment, the identity of any other drugs being administered to the animal, the age, size and species of the animal, and like factors well known in the medical and veterinary arts.
  • a suitable daily dose will be that amount which is the lowest dose effective to produce a therapeutic effect.
  • the total daily dosage will be determined by an attending physician or veterinarian within the scope of sound medical judgment.
  • the effective daily dose of a compound of formula (1), or a pharmaceutically-acceptable salt thereof maybe administered as two, three, four, five, six or more sub-doses, administered separately at appropriate intervals throughout the day.
  • Treatment of a ⁇ -lactam- antibiotic-resistant bacterial infection according to the invention includes mitigation, as well as elimination, of the infection.
  • Animals treatable according to the invention include mammals. Mammals treatable according to the invention include dogs, cats, other domestic animals, and humans.
  • Compounds of formula (1) or pharmaceutically-acceptable salts thereof may be administered to an animal patient for therapy by any suitable route of administration, including orally, nasally, rectally, intravaginally, parenterally, intracisternally and topically, as by powders, ointments or drops, including buccally and sublingually.
  • suitable routes of administration including orally, nasally, rectally, intravaginally, parenterally, intracisternally and topically, as by powders, ointments or drops, including buccally and sublingually.
  • the preferred routes of administration are orally and parenterally.
  • compositions comprise the active ingredient(s) in admixture with one or more pharmaceutically-acceptable earners and, optionally, with one or more other compounds, drugs or other materials.
  • Each carrier must be "acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • compositions of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. Regardless of the route of administration selected, the active ingredient(s) are formulated into pharmaceutically-acceptable dosage forms by conventional methods known to those of skill in the art.
  • the amount of the active ingredient(s) which will be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration and all of the other factors described above.
  • the amount of the active ingredient(s) which will be combined with a carrier material to produce a single dosage form will generally be that amount of the active ingredient(s) which is the lowest dose effective to produce a therapeutic effect.
  • Methods of preparing pharmaceutical formulations or compositions include the step of bringing the active ingredient(s) into association with the carrier and, optionally, one or more accessory ingredients.
  • the formulations are prepared by uniformly and intimately bringing the active ingredient(s) into association with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
  • Formulations of the invention 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 non-aqueous 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) and/or as mouth washes and the like, each containing a predetermined amount of the active ingredient(s).
  • the active ingredient(s) may also be administered as a bolus, electuary or paste.
  • the active ingredient(s) is/are mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pynolidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) abso ⁇ tion accelerators, such as quaternary ammonium compounds; (7) we
  • 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.
  • 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 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 powdered active ⁇ ngredient(s) moistened with an inert liquid diluent.
  • the tablets, and other solid dosage forms ot the pharmaceutical compositions of the present invention may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient(s) therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter.
  • compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner.
  • opacifying agents include polymeric substances and waxes.
  • the active ingredient(s) can also be in microencapsulated form.
  • Liquid dosage forms for oral administration of the active ingredient(s) include pharmaceutically-acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • 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, and mixtures thereof.
  • the oral compositions can also include adjuvants such as we
  • Suspensions in addition to the active ingredient(s), may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.
  • suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.
  • Formulations of the pharmaceutical compositions of the invention for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing the active ingredient(s) with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or salicylate and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active ingredient(s).
  • suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or salicylate and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active ingredient(s).
  • Formulations of the present invention 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.
  • Dosage forms for the topical or transdermal administration of the active ingredient(s) include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.
  • the active ingredient(s) may be mixed under sterile conditions with a pharmaceutically-acceptable carrier, and with any buffers, or propellants which may be required.
  • the ointments, pastes, creams and gels may contain, in addition to the active ingredient(s), 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.
  • 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.
  • Powders and sprays can contain, in addition to the active ingredient(s), excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances.
  • Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.
  • Transdermal patches have the added advantage of providing controlled delivery of the active ingredient(s) to the body. Such dosage forms can be made by dissolving, dispersing or otherwise inco ⁇ orating the active ingredient(s) in a proper medium, such as an elastomeric matrix material. Abso ⁇ tion enhancers can also be used to increase the flux of the active ingredient(s) across the skin. The rate of such flux can be controlled by either providing a rate-controlling membrane or dispersing the active ingredient(s) in a polymer matrix or gel.
  • compositions of this invention suitable for parenteral administration comprise the active ingredient(s) in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous 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, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
  • aqueous and nonaqueous carriers examples 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.
  • polyols such as glycerol, propylene glycol, polyethylene glycol, and the like
  • vegetable oils such as olive oil
  • injectable organic esters such as ethyl oleate.
  • Proper fluidity can 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.
  • compositions may also contain adjuvants such as wetting agents, emulsifying agents and dispersing agents. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like in the compositions.
  • isotonic agents such as sugars, sodium chloride, and the like in the compositions.
  • prolonged abso ⁇ tion of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay abso ⁇ tion such as aluminum monostearate and gelatin.
  • Injectable depot forms are made by forming microencapsule matrices of the active ingredient(s) in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of the active ingredient(s) to polymer, and the nature of the particular polymer employed, the rate of release of the active ingredient(s) can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the active ingredient(s) in liposomes or microemulsions which are compatible with body tissue. The injectable materials can be sterilized for example, by filtration through a bacterial-retaining filter.
  • the formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a lyophilized condition requiring only the addition of the sterile liquid carrier, for example water for injection, immediately prior to use.
  • sterile liquid carrier for example water for injection
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the type described above.
  • compositions of the present invention may also be used in the form of veterinary formulations, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules or pellets for admixture with feed stuffs, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular or intravenous injection as, for example, a sterile solution or suspension or, when appropriate, by intramammary injection where a suspension or solution is introduced into the udder of the animal via its teat; (3) topical application, for example, as a cream, ointment or spray applied to the skin; or (4) intravaginally, for example, as a pessary, cream or foam.
  • oral administration for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules or pellets for admixture with feed
  • Compound 1 has been also synthesized by traditional liquid phase method and in larger amount.
  • 0.3 g (1.61 mmol) of 3-aminophenylboronic acid hemisulfate was dissolved in chloroform (CHC1 3 , 30 mL) followed by pH adjustment to 10 using sodium hydroxide (4 ⁇ NaOH).
  • 4-(benzene- sulfonyl)-thiophene-2-sulphonyl chloride 1.5 eq; 2.415 mmol, 0.779 g
  • acetone 10 mL
  • 3-(3 , 5-Dimethyl-isoxazole-4-sulfonylamino)-phenyl boronic acid (2) 0.2 g ( 1.075 mmol) of 3APB was dissolved in sodium bicarbonate (0.5 M, NaHCO 3 , 20 mL), followed by pH adjustment to 10 using sodium hydroxide (4N NaOH). To the solution, kept under stirring at 30 ° C, was then added 3 , 5-dimethyl-isoxazole-4-sulphonyl chloride ( 1.5 eq; 1.61 mmol, 0.315 g) already dissolved in acetone (10 mL).
  • Compound 3 was prepared according to the method described for 2 starting from 0.2 g (1.075 mmol) of 3 APB and 5-benzenesulfonyl-thiophene-2-sulfonyl chloride (1.5 eq; 1.61 mmol, 0.521 g). Yield 0.140 g, 31%.
  • AmpC was expressed in E. coli JM109 cells (available from American Type Culture Collection, Rockville, MD, accession no. 53323) in which the native AmpC gene was attenuated or completely removed (obtained from Larry Blaszczak, Eli Lilly and Co., Indianapolis, Indiana) as described in Usher etal, Biochemistry, 37, 16082-16092 (1998). Briefly, DNA coding for the enzyme was located on a plasmid under the control of a temperature sensitive repressor. Cells containing this plasmid were grown in 2 liters of LB broth in a fermentor to log phase. Enzyme expression was then induced by temperature shock, and the cells were allowed to grow overnight.
  • AmpC protein was purified from the supernatant over an Affigel-10 aminophenyl boronate affinity column (Bio-Rad Laboratories, 1000 Alfred Nobel Drive, Hercules, CA). The purity of the sample was estimated by HPLC to be 96% or better. The amount of enzyme produced was estimated to be 150 m? based on absorbance at 280 nm.
  • TEM-1 was provided by Natalie Strynadka, University Of Alberta, Edmonton, Canada. Alternatively, TEM-1 may be produced as follows. The TEM-1 gene is cloned into Hpal site of pALTER-EX2 (Promega). The gene is under control of the T7 promoter which is turned on for protein expression.
  • TEM-1 may be expressed in JM109 cells, as well as several other E. coli strains. Cells are grown to late log phase, followed by induction of protein expression. The cells are spun down and the supernatant, into which the enzyme has been exported, is collected. Because the enzyme has been exported into the supernatant, purification may be achieved using standard column chromatography, as described in Matagne et al. Biochem J. 265, 131-146 (1990); Escobar et al, Biochemistry 33, 7619-7626 (1994).
  • Enzymatic testing was typically started at an upper concentration limit determined by the solubility and absorbance profile of the compound.
  • the background rate of cephalothin hydrolysis under these conditions was found to be two to three orders of magnitude less than the rate of the enzyme-mediated cephalothin hydrolysis, so no corcection for background hydrolysis of substrate was used.
  • TEM-1 100 ⁇ M 6- ⁇ - furylacryloylamidopenicillanic acid, triethylammonium salt (FAP), was used as the substrate, the reaction was monitored at 340 nm (FAP ⁇ -lactam absorbance peak) and the cycle time was increased to 25 seconds (since this substrate was somewhat light sensitive). Due to the light sensitivity of FAP, the background rate of hydrolysis for this substrate was found to be minimal, but not insignificant, so all measured control and inhibited cell rates were corrected by subtraction of the FAP background rate. All other conditions for the TEM-1 assays were identical to those for the AmpC assays. DMSO was added to enzyme controls in all cases.
  • FAP triethylammonium salt
  • Standard 1 mm path length quartz spectrophotometric cells (Hellma Cells, Inc., Jamaica, NY) were used in the assays. All assays were performed on the same HP8543 spectrophotometer noted earlier. Linear and quadratic fits to the absorbance data for the full time course of each reaction were used to determine the reaction rate for each spectrophotometric cell. The resulting reaction rate data were used to calculate the inhibition constants for each potential inhibitor using the method of Waley (S.G. Waley, Biochem. J. 205, 631-633 (1982)). Briefly, this method involves the use of the integrated Michaelis-Menten equation to calculate Kj values for enzyme inhibitors from a comparison of the reaction rates of uninhibited and inhibited enzymatic reactions.
  • elastase substrate 1 elastase substrate 1, N ⁇ - methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide, was purchased from Calbiochem, San Diego, CA. All enzymes used for specificity testing were purchased from Sigma Chemical, St. Louis, MO.
  • ⁇ -chymotrypsin 3 ⁇ l of a 1 mg/ml enzyme stock solution (50 mM phosphate buffer, pH 7) was incubated with the boronate being tested for 5 minutes; then the reaction was initialized by addition of 630 ⁇ M BTEE from a DMSO stock solution. The reaction was performed at 25 °C and monitored at 260 nm.
  • ⁇ - trypsin 40 ⁇ l of a 0.8 mg/ml enzyme stock solution (50 mM phosphate buffer, pH 7) was incubated with the boronate being tested for 5 minutes; then the reaction was initialized by addition of 600 ⁇ M BAEE from a DMSO stock solution.
  • elastase 50 ⁇ l of a 1 mg/ml enzyme stock solution (50 mM phosphate buffer, pH 7) was incubated with the boronate being tested for 5 minutes; then the reaction was initialized by addition of 64 ⁇ M elastase substrate 1 from a DMSO stock solution.
  • Tables 1 A and IB The results of the testing are presented in Tables 1 A and IB below. Certain prior art compounds (Table 1 A) were tested for comparative purposes. All of these compounds were obtained from Lancaster Synthesis, Windham, NH, except for MAPB, which was obtained from Sigma, St. Louis, MO. All of these compounds were used as is with no additional purification or verification performed. The compounds listed in Table IB were synthesized as described in Examples 1 and 2.
  • NSULFB /w-(dansylamidophenyl)-boronic acid
  • 4FORMB 4-formylphenylboronate
  • 4MEPB 4-methylphenylboronate
  • MAPB /w-aminophenylboronate
  • 2FDB phenylboronic acid
  • 2FORMB 2-formylphenylboronate.
  • Bacterial cell culture testing was performed and interpreted following the guidelines of the National Committee for Clinical Laboratory Standards (National Committee for Clinical Laboratory Standards. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically. Approved Standard M7-A3. National Committee for Clinical Laboratory Standards, Villanova, Pa. 1993).
  • the tested bacterial strains were all clinical isolates from the Hospital Ramon y Cajal in Madrid, and all of the strains listed in Table 2 below are available from Jesus Blazquez and Fernando Baquero, Servicio deMicrobiologia, Hospital Ramon y Cajal, National Institute ofHealth, Madrid, Spain. After incubation, the growth of the cells was visually inspected. The minimum inhibitory concentration (MIC) is the lowest concentration ( ⁇ g/ml) where no cell growth was observed. The results are presented in Table 2 below.
  • Amx amoxicillin
  • Clav Clavulanic acid
  • Amoxicillin/lnhibitor 1/3 (w/w)
  • Amoxicillin/Clav 2 1 (w/w)
  • the tested bacterial strains do not express class C (AmpC-like) ⁇ -lactamases (see Example 3). Instead, they express class A ⁇ -lactamases, most probably an enzyme that resembles the PCI ⁇ -lactamase from St ⁇ phlococcus aureus. Curiously, the tested bacteria are much more affected by these inhibitors than most bacteria expressing AmpC-like enzymes (data not shown). This probably reflects the fact that Gram positive bacteria lack an outer membrane, apparently a feature of Gram negative bacteria that seriously diminishes efficacy against them, and only Gram negative bacteria are known to express class C ⁇ -lactamases. In view of the results in Table 2, compounds 1, 15, and 16, currently the best inhibitors, were tested against the PCI ⁇ -lactamase of S.

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Abstract

L'invention concerne de nouveaux inhibiteurs non β-lactame de β-lactamases. Elle concerne plus particulièrement des inhibiteurs qui sont des acides boroniques représentés par la formule (1) définie dans les spécifications. Ces composés peuvent être utilisés avec des β-lactamines dans le traitement d'infections bactériennes résistant aux β-lactamines. L'invention se rapporte enfin à une composition pharmaceutique comprenant ces composés.
PCT/US1999/029958 1998-12-16 1999-12-16 INHIBITEURS DE β-LACTAMASES ET LEUR UTILISATION WO2000035904A1 (fr)

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US6858592B2 (en) 2001-06-29 2005-02-22 Genzyme Corporation Aryl boronic acids for treating obesity
US7041280B2 (en) 2001-06-29 2006-05-09 Genzyme Corporation Aryl boronate functionalized polymers for treating obesity
US7112572B2 (en) 2002-09-09 2006-09-26 Trigen Limited Multivalent metal salts of boronic acids
US9012491B2 (en) 2011-08-31 2015-04-21 Rempex Pharmaceuticals, Inc. Heterocyclic boronic acid ester derivatives and therapeutic uses thereof
US9101638B2 (en) 2013-01-04 2015-08-11 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9132140B2 (en) 2013-01-04 2015-09-15 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9156858B2 (en) 2012-05-23 2015-10-13 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9241947B2 (en) 2013-01-04 2016-01-26 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9296763B2 (en) 2010-08-10 2016-03-29 Rempex Pharmaceuticals, Inc. Cyclic boronic acid ester derivatives and therapeutic uses thereof
US9642869B2 (en) 2013-01-04 2017-05-09 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9687497B1 (en) 2014-05-05 2017-06-27 Rempex Pharmaceuticals, Inc. Salts and polymorphs of cyclic boronic acid ester derivatives and therapeutic uses thereof
US9963467B2 (en) 2014-05-19 2018-05-08 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US10206937B2 (en) 2014-07-01 2019-02-19 Qpex Biopharma, Inc. Boronic acid derivatives and therapeutic uses thereof
US10294249B2 (en) 2016-06-30 2019-05-21 Qpex Biopharma, Inc. Boronic acid derivatives and therapeutic uses thereof
US10385074B2 (en) 2014-05-05 2019-08-20 Rempex Pharmaceuticals, Inc. Synthesis of boronate salts and uses thereof
US10561675B2 (en) 2012-06-06 2020-02-18 Rempex Pharmaceuticals, Inc. Cyclic boronic acid ester derivatives and therapeutic uses thereof
US10618918B2 (en) 2015-03-17 2020-04-14 Qpex Biopharma, Inc. Substituted boronic acids as antimicrobials
US10662205B2 (en) 2014-11-18 2020-05-26 Qpex Biopharma, Inc. Cyclic boronic acid ester derivatives and therapeutic uses thereof
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5612378A (en) * 1995-06-06 1997-03-18 3-Dimensional Pharmaceuticals, Inc. Bis-arylsulfonylaminobenzamide derivatives and the use thereof as factor Xa inhibitors
US6008377A (en) * 1996-03-19 1999-12-28 Eli Lilly And Company Synthesis of 3-[4-(2-aminoethoxy)-benzoyl]-2-aryl-6-hydroxy-benzo[B]thiophenes

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5612378A (en) * 1995-06-06 1997-03-18 3-Dimensional Pharmaceuticals, Inc. Bis-arylsulfonylaminobenzamide derivatives and the use thereof as factor Xa inhibitors
US6008377A (en) * 1996-03-19 1999-12-28 Eli Lilly And Company Synthesis of 3-[4-(2-aminoethoxy)-benzoyl]-2-aryl-6-hydroxy-benzo[B]thiophenes

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US7041280B2 (en) 2001-06-29 2006-05-09 Genzyme Corporation Aryl boronate functionalized polymers for treating obesity
US7049304B2 (en) 2001-06-29 2006-05-23 Genzyme Corporation Aryl boronic acids for treating obesity
US6858592B2 (en) 2001-06-29 2005-02-22 Genzyme Corporation Aryl boronic acids for treating obesity
US7371729B2 (en) 2002-09-09 2008-05-13 Trigen Limited Boronic acid salts useful in parenteral formulations
US7112572B2 (en) 2002-09-09 2006-09-26 Trigen Limited Multivalent metal salts of boronic acids
US9296763B2 (en) 2010-08-10 2016-03-29 Rempex Pharmaceuticals, Inc. Cyclic boronic acid ester derivatives and therapeutic uses thereof
US11684629B2 (en) 2010-08-10 2023-06-27 Melinta Subsidiary Corp. Therapeutic uses of pharmaceutical compositions comprising cyclic boronic acid ester derivatives
US11090319B2 (en) 2010-08-10 2021-08-17 Melinta Subsidiary Corp. Therapeutic uses of pharmaceutical compositions comprising cyclic boronic acid ester derivatives
US10183034B2 (en) 2010-08-10 2019-01-22 Rempex Pharmaceuticals, Inc. Therapeutic uses of pharmaceutical compositions comprising cyclic boronic acid ester derivatives
US10639318B2 (en) 2010-08-10 2020-05-05 Rempex Pharmaceuticals, Inc. Therapeutic uses of pharmaceutical compositions comprising cyclic boronic acid ester derivatives
US9694025B2 (en) 2010-08-10 2017-07-04 Rempex Pharmaceuticals, Inc. Cyclic boronic acid ester derivatives and therapeutic uses thereof
US10004758B2 (en) 2010-08-10 2018-06-26 Rempex Pharmaceuticals, Inc. Cyclic boronic acid ester derivatives and methods of making the same
US10172874B2 (en) 2010-08-10 2019-01-08 Rempex Pharmaceuticals, Inc. Pharmaceutical compositions comprising cyclic boronic acid ester derivatives
US9012491B2 (en) 2011-08-31 2015-04-21 Rempex Pharmaceuticals, Inc. Heterocyclic boronic acid ester derivatives and therapeutic uses thereof
US9156858B2 (en) 2012-05-23 2015-10-13 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US11007206B2 (en) 2012-06-06 2021-05-18 Melinta Subsidiary Corp. Cyclic boronic acid ester derivatives and therapeutic uses thereof
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US9132140B2 (en) 2013-01-04 2015-09-15 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9642869B2 (en) 2013-01-04 2017-05-09 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9101638B2 (en) 2013-01-04 2015-08-11 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9241947B2 (en) 2013-01-04 2016-01-26 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US9687497B1 (en) 2014-05-05 2017-06-27 Rempex Pharmaceuticals, Inc. Salts and polymorphs of cyclic boronic acid ester derivatives and therapeutic uses thereof
US10669292B2 (en) 2014-05-05 2020-06-02 Rempex Pharmaceuticals, Inc. Synthesis of boronate salts and uses thereof
US10385074B2 (en) 2014-05-05 2019-08-20 Rempex Pharmaceuticals, Inc. Synthesis of boronate salts and uses thereof
US9963467B2 (en) 2014-05-19 2018-05-08 Rempex Pharmaceuticals, Inc. Boronic acid derivatives and therapeutic uses thereof
US10206937B2 (en) 2014-07-01 2019-02-19 Qpex Biopharma, Inc. Boronic acid derivatives and therapeutic uses thereof
US10662205B2 (en) 2014-11-18 2020-05-26 Qpex Biopharma, Inc. Cyclic boronic acid ester derivatives and therapeutic uses thereof
US10618918B2 (en) 2015-03-17 2020-04-14 Qpex Biopharma, Inc. Substituted boronic acids as antimicrobials
US10570159B2 (en) 2016-06-30 2020-02-25 Qpex Biopharma, Inc. Boronic acid derivatives and therapeutic uses thereof
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