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WO2018162928A1 - Procédé de restauration de l'efficacité d'un agent antibactérien - Google Patents

Procédé de restauration de l'efficacité d'un agent antibactérien Download PDF

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Publication number
WO2018162928A1
WO2018162928A1 PCT/GB2018/050613 GB2018050613W WO2018162928A1 WO 2018162928 A1 WO2018162928 A1 WO 2018162928A1 GB 2018050613 W GB2018050613 W GB 2018050613W WO 2018162928 A1 WO2018162928 A1 WO 2018162928A1
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Prior art keywords
antibiotic resistance
methyl
antibacterial agent
quinoline
polymyxin
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PCT/GB2018/050613
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English (en)
Inventor
Anthony Coates
Yanmin Hu
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Helperby Therapeutics Ltd
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Helperby Therapeutics Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/50—Cyclic peptides containing at least one abnormal peptide link
    • C07K7/54—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring
    • C07K7/60—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring the cyclisation occurring through the 4-amino group of 2,4-diamino-butanoic acid
    • C07K7/62—Polymyxins; Related peptides
    • 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
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • 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
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/137—Arylalkylamines, e.g. amphetamine, epinephrine, salbutamol, ephedrine or methadone
    • 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
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/138—Aryloxyalkylamines, e.g. propranolol, tamoxifen, phenoxybenzamine
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/336—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having three-membered rings, e.g. oxirane, fumagillin
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/407—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/542—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/545—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A61K31/7072—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/12—Cyclic peptides, e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca

Definitions

  • the present invention relates to a method for restoring the efficacy or prolonging the life of an antibacterial agent by combining the antibacterial agent with at least two antibiotic resistance breakers, wherein each antibiotic resistance breaker is used independently with the antibacterial agent so as to at least partially restore the efficacy thereof and wherein each antibiotic resistance breaker has a different mechanism of action.
  • the invention also provides the use of at least two antibiotic resistance breakers to restore efficacy or prolong the life of an antibacterial agent.
  • ARBs antibiotic resistance breakers
  • ⁇ -lactamase inhibitors such as clavulanic acid
  • ⁇ -lactam antibiotics such as amoxicillin
  • ⁇ - ⁇ 3 ⁇ 3 ⁇ / ⁇ - ⁇ 3 ⁇ 3 ⁇ 33 ⁇ inhibitor combinations include Tazocin® (piperacillin/tazobactam), Avycaz® (ceftazidime/avibactam) and CarbA® (meropenem/vaborbactam). It would, however, be preferable to identify ARBs which restore one or more key members of each mechanistic antibiotic class, for example those antibiotics used against Gram-negative bacteria. According to Nature Reviews, Drug Discovery, 14, 821-832 (2015), the antibiotics that most need ARBs are:
  • cephalosporins and carbapenems cephalosporins and carbapenems
  • polymyxins are of particular interest because polymyxin E (colistin) is one of the agents currently used to combat bacteria that are resistant to the strongest antibiotics.
  • Colistin is commercially available in Europe under the trade name Colomycin® in intravenous form.
  • Intravenous Colomycin® includes colistimethate sodium which undergoes hydrolysis to the active substance colistin in aqueous solution and is indicated for the treatment of serious infections due to selected aerobic Gram-negative pathogens in patients with limited treatment options.
  • WO2012032360 discloses a combination comprising phenoxybenzamine or a pharmaceutically acceptable derivative thereof and a polymyxin selected from polymyxin E and polymyxin B or a pharmaceutically acceptable derivative thereof, and its use in treating a microbial infection.
  • International Patent Application, Publication Number WO2014147405 discloses the use of colistin (polymyxin E) in combination with zidovudine for treating a microbial infection.
  • International Patent Application, Publication Number WO2016097754 discloses a combination comprising suloctidil or a pharmaceutically acceptable derivative or prodrug thereof, and a polymyxin selected from polymyxin E and polymyxin B or a pharmaceutically acceptable derivative thereof, and its use in treating a microbial infection.
  • an antibacterial agent e.g. colistin
  • antibiotic resistance breaker will also, however, inevitably occur.
  • the present invention is therefore based on the unexpected finding that the efficacy of an antibacterial agent can be prolonged and/or restored by using at least two antibiotic resistance breakers with different mechanisms of action for at least two time periods: the first antibiotic resistance breaker being combined with the antibacterial agent for a first time period, and the second antibiotic resistance breaker being combined with the antibacterial agent for a second time period.
  • Effectiveness in the context of the present invention refers to the antibacterial activity of the antibacterial agent against a particular bacterium, preferably a Gram-negative bacterium.
  • a Gram-negative bacterium such as E.coli or K. pneumoniae, preferably a drug-resistant strain of E.coli or K.pneumoniae.
  • the antibacterial activity of the antibacterial agent and the antibiotic resistance breaker against the bacterium of interest is additive or synergistic.
  • the antibacterial activity is synergistic. Synergy in the context of antibacterial drugs is measured in a number of ways that conform to the generally accepted opinion that "synergy is an effect greater than additive".
  • FICI fractional inhibitory concentration index
  • Orhan et al J. Clin. Microbiol. 2005, 43(1): 140 describes the chequerboard method and analysis in the paragraph bridging pages 140-141 , and explains that the FICI value is a ratio of the sum of the MIC (Minimum Inhibitory Concentration) level of each individual component alone and in the mixture.
  • the combination is considered synergistic when the ⁇ FIC is ⁇ 0.5, indifferent when the ⁇ FIC is >0.5 but ⁇ 2, and antagonistic when the ⁇ FIC is ⁇ 2.
  • a first aspect of the invention is a method for restoring efficacy of an antibacterial agent.
  • the method comprises (i) combining the antibacterial agent with a first antibiotic resistance breaker after a first time period; and (ii) combining the antibacterial agent with a second antibiotic resistance breaker after a second time period.
  • the efficacy of the antibacterial agent decreases due to increasing antibiotic resistance
  • each antibiotic resistance breaker at least partially restores the efficacy of the antibacterial agent relative to the end of the preceding time period.
  • the first and second antibiotic resistance breakers have different mechanisms of action.
  • the invention provides the use of at least two antibiotic resistance breakers to prolong the efficacy of an antibacterial agent, where the at least two antibiotic resistance breakers have different mechanisms of action.
  • Figure 1 is a graph of relative efficacy of antibiotic therapy against an approximate historic and anticipated timeline. This graph portrays the invention in a schematic manner showing how antibiotic resistance breakers (ARBs) can be combined with an antibacterial agent after successive time periods so as to at least partially restore the efficacy of the antibacterial agent.
  • ARBs antibiotic resistance breakers
  • Figure 2 contains images of E.coli tolC cells showing the characteristic phenotype produced by treatment with the compounds in Example 3. Each image is entitled with the compound tested and the concentration used (in ⁇ g/ml).
  • Figure 2A cell membranes are shown in red, DNA is shown in blue and green staining indicates permeabilization of the cell membrane.
  • White scale bar is 1 ⁇ .
  • Figure 2B is the greyscale version of Figure 2A.
  • Figure 3 contains images of E.coli ATCC 25922 cells showing the predominant phenotypes produced by treatment in Example 3 with colistin and/or mefloquine for 2 hours at the concentrations indicated ⁇ g/ml).
  • cell membranes are shown in red, DNA in blue and green staining indicates permeabilization of the cell membrane.
  • White scale bar is 1 ⁇ .
  • Figure 3B is the greyscale version of Figure 3A.
  • Figure 4 is a chart which shows the viable cell counts for E.coli ATCC 25922 treated with DMSO, colistin, mefloquine or a combination of colistin and mefloquine at 0, 2 and 4 hours. Concentrations are in ⁇ g/ml.
  • Figure 5 contains images of E.coli ATCC 25922 cells showing the predominant phenotypes produced by treatment in Example 3 with colistin and/or suloctidil for 2 hours at the concentrations indicated ⁇ g/ml).
  • cell membranes are shown in red, DNA in blue and green staining indicates permeabilization of the cell membrane.
  • White scale bar is 1 ⁇ .
  • Figure 5B is the greyscale version of Figure 5A.
  • Figure 6 is a chart which shows the viable cell counts for E.coli ATCC 25922 treated with DMSO, colistin, suloctidil or a combination of colistin and suloctidil at 0, 2 and 4 hours. Concentrations are in ⁇ .
  • Figure 7 contains images of E.coli ATCC 25922 cells showing the predominant phenotypes produced by treatment in Example 3 with colistin and/or zidovudine for 2 hours at the concentrations indicated ⁇ g/ml).
  • cell membranes are shown in red, DNA in blue and green staining indicates permeabilization of the cell membrane.
  • White scale bar is 1 ⁇ .
  • Figure 7B is the greyscale version of Figure 7A.
  • Figure 8 is a chart which shows the viable cell counts for E.coli ATCC 25922 treated with DMSO, colistin, zidovudine or a combination of colistin and zidovudine at 0, 2 and 4 hours. Concentrations are in ⁇ g/ml.
  • Figure 9 contains images of E.coli ATCC 25922 cells showing the predominant phenotypes produced by treatment in Example 3 with colistin and/or phenoxybenzamine for 2 hours at the concentrations indicated ⁇ g/ml).
  • cell membranes are shown in red, DNA in blue and green staining indicates permeabilization of the cell membrane.
  • White scale bar is 1 ⁇ .
  • Figure 9B is the greyscale version of Figure 9A.
  • Figure 10 is a chart which shows the viable cell counts for E.coli ATCC 25922 treated with DMSO, colistin, phenoxybenzamine or a combination of colistin and phenoxybenzamine at 0, 2 and 4 hours. Concentrations are in ⁇ g/ml.
  • Figure 11 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 0.5% DMSO for 30 minutes (left) and 120 minutes (right). In Figure 11A cell membranes are shown in red and DNA in blue. The green staining (SYTOX Green) did not produce any visible results. White scale bar is 1 ⁇ .
  • Figure 1 1 B is the greyscale version of Figure 1 1A.
  • Figure 12 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 0.06 ⁇ g/ml azidothymidine (zidovudine) for 30 minutes (left) and 120 minutes (right).
  • azidothymidine azidovudine
  • Figure 12A cell membranes are shown in red and DNA in blue. The green staining (SYTOX Green) did not produce any visible results.
  • White scale bar is 1 ⁇ .
  • Figure 12B is the greyscale version of Figure 12A.
  • Figures 13A and 13B are the same as Figures 12A and 12B except treatment is with 0.3 ⁇ g/ml azidothymidine (zidovudine).
  • Figure 14 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 0.002 ⁇ g/ml ciprofloxacin for 30 minutes (left) and 120 minutes (right).
  • cell membranes are shown in red and DNA in blue. The green staining (SYTOX Green) did not produce any visible results.
  • White scale bar is 1 ⁇ .
  • Figure 14B is the greyscale version of Figure 14A.
  • Figures 15A and 15B are the same as Figures 14A and 14B except treatment is with 0.01 ⁇ g/ml ciprofloxacin.
  • Figure 16 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 20 ⁇ g/ml cephalexin for 30 minutes (left) and 120 minutes (right).
  • Figure 16A cell membranes are shown in red and DNA in blue. The green staining (SYTOX Green) did not produce any visible results.
  • White scale bar is 1 ⁇ .
  • Figure 16B is the greyscale version of Figure 16A.
  • Figure 17 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 25 ⁇ g/ml cerulenin for 120 minutes (left) and 240 minutes (right).
  • Figure 17A cell membranes are shown in red and DNA in blue.
  • the green staining (SYTOX Green) indicates permeabilization of the cell membrane.
  • White scale bar is 1 ⁇ .
  • Figure 17B is the greyscale version of Figure 17A.
  • Figure 18 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 7.5 ⁇ g/ml rifampicin for 30 minutes (left) and 120 minutes (right).
  • Figure 18A cell membranes are shown in red and DNA in blue. The green staining (SYTOX Green) did not produce any visible results.
  • White scale bar is 1 ⁇ .
  • Figure 18B is the greyscale version of Figure 18A.
  • Figure 19 is a clustergram of azidothymine (zidovudine) and the different classes of antibiotics with E.coli tolC treated with 5X MIC for 120 minutes (or 240 minutes for cerulenin).
  • Figure 20 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 1.5 ⁇ g/ml daunorubicin for 30 minutes (left) and 120 minutes (right).
  • cell membranes are shown in red and DNA in blue. The green staining (SYTOX Green) did not produce any visible results.
  • White scale bar is 1 ⁇ .
  • Figure 20B is the greyscale version of Figure 20A.
  • Figures 21 A and 21 B are the same as Figures 20A and 20B except treatment is with 7.5 ⁇ g/ml daunorubicin.
  • the green staining indicates permeabilization of the cell membrane.
  • Figure 22 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 1 ⁇ g/ml novobiocin for 30 minutes (left) and 120 minutes (right).
  • cell membranes are shown in red and DNA in blue.
  • the green staining (SYTOX Green) did not produce any visible results.
  • White scale bar is 1 ⁇ .
  • Figure 22B is the greyscale version of Figure 22A.
  • Figures 23A and 23B are the same as Figures 22A and 22B except treatment is with 5 ⁇ g/ml novobiocin.
  • Figure 24 contains images of E.coli tolC showing the predominant phenotypes produced by treatment with 0.03 ⁇ g/ml mitomycin C for 30 minutes (left) and 120 minutes (right).
  • cell membranes are shown in red and DNA in blue.
  • the green staining SYTOX Green
  • White scale bar is 1 ⁇ .
  • Figure 24B is the greyscale version of Figure 24A.
  • Figures 25A and 25B are the same as Figures 24A and 24B except treatment is with 0.15 ⁇ g/ml mitomycin C.
  • Figure 26 is a clustergram of azidothymine (zidovudine) and the DNA replication inhibitors with E.coli tolC treated with 5X MIC for 120 minutes.
  • cy or cyclo refers to the cyclic part of the peptide, enclosed within brackets; Dab or Dbu refers to ⁇ , ⁇ -diamino-n-butyryl (i.e. 2,4-diaminobutyryl); Abu refers to 2-aminobutyryl; Thr refers to L-threonine; DThr refers to D-threonine; DPhe refers to D- phenylamine; Leu refers to L-leucine; DSer refers to D-serine and OA refers to octanoyl.
  • a first aspect of the invention relates to a method for restoring efficacy of an antibacterial agent as defined herein.
  • Effectiveness in the context of the present invention refers to the antibacterial activity of the antibacterial agent against a particular bacterium, preferably a Gram-negative bacterium.
  • the antibacterial activity of the antibacterial agent and the antibiotic resistance breaker is additive (indifferent) or synergistic.
  • the antibacterial activity is synergistic.
  • the antibacterial activity is against a drug-resistant Gram-negative bacterium such as E.coli or K.pneumoniae.
  • the method further comprises (iii) combining the antibacterial agent with a third antibiotic resistance breaker after a third time period, where during the third time period the efficacy of the antibacterial agent decreases due to increasing antibiotic resistance, and the third antibiotic resistance breaker at least partially restores the efficacy of the antibacterial agent relative to the end of the second time period.
  • the third antibiotic resistance breaker preferably has a different mechanism of action to both the first and second antibiotic resistance breakers.
  • the method comprises (iv) combining the antibacterial agent with a fourth antibiotic resistance breaker after a fourth time period, where during the fourth time period, the efficacy of the antibacterial agent decreases due to increasing antibiotic resistance, and the fourth antibiotic resistance breaker at least partially restores the efficacy of the antibacterial agent relative to the end of the third time period.
  • the fourth antibiotic resistance breaker preferably has a different mechanism of action to each of the first, second and third antibiotic resistance breakers.
  • antibiotic resistance refers to the ability of bacteria and other microorganisms to resist the effects of an antibiotic to which they were once sensitive. The antibiotic becomes ineffective against the bacteria. When the bacteria become resistant to most antibacterial agents, they are often referred to in the art as "superbugs".
  • Efflux pumps for example are high-affinity reverse transport systems located in the membrane that transport the antibiotic out of the cell.
  • a specific enzyme which modifies the antibiotic in a way that it loses its activity or degrades the antibiotic so it becomes inactive.
  • Bacteria may alternatively be inherently resistant to an antibiotic.
  • an organism may lack a transport system for an antibiotic, or the target for the antibiotic molecule, or as in the case of Gram-negative bacteria, the cell wall may be covered with an outer membrane that establishes a permeability barrier against the antibiotic.
  • antibiotic resistance breaker refers to non-antibiotic compounds that, when combined with existing antibiotics, act to block resistance or enhance antibacterial activity. These compounds are also known in the art as “antibiotic adjuvants", “resistance breakers” and “antibiotic potentiators”. They may be known compounds (repurposed existing drugs) or new chemical entities.
  • at least one of the antibiotic resistance breakers of the present invention is a repurposed existing drug.
  • each of the antibiotic resistance breakers is a repurposed existing drug.
  • One advantage of using a repurposed drug is that these drugs have known toxicology and pharmacology profiles, and therefore can lead to considerable cost savings by eliminating much of the toxicological and pharmacokinetic assessment that would normally be required for approval of a new drug.
  • the antibiotic resistance breakers in the method or use of the present invention may, for example, be selected from the group consisting of zidovudine, suloctidil, mefloquine, phenoxybenzamine or pharmaceutically acceptable derivatives thereof. All of these compounds are known non-antibiotic drugs and in the context of the present invention are repurposed existing drugs.
  • Zidovudine is also known as azidothymidine (AZT) and is an anti retroviral medication used to prevent and treat HIV/AIDs. It is generally recommended for use with other antiretrovirals and has the following chemical structure:
  • lUPAC name is 1-[(2/ : ?,4S,5S)-4-Azido-5-(hydroxymethyl)oxolan-2-yl]-5-methylpyrimidine- 2,4-dione, and it is available by prescription only under the trade name Retrovir®.
  • Suloctidil is a sulfur-containing amino alcohol that was brought to market in the early 1970s as a vasodilator. It has the following chemical structure:
  • Mefloquine is an orally administered medication used in the prevention and treatment of malaria. It is commercially available in Europe under the trade name Lariam® in tablet form. Lariam® tablets include the hydrochloride salt of mefloquine and are indicated for the therapy and prophylaxis of malaria.
  • Phenoxybenzamine is a non-selective, irreversible alpha blocker which is marketed under the trade name Dibenzyline®. It is used in the treatment of hypertension and has the following chemical structure:
  • the first antibiotic resistance breaker is zidovudine or a pharmaceutically acceptable derivative thereof.
  • the second, third and/or fourth antibiotic resistance breakers are selected from the group consisting of suloctidil, mefloquine and phenoxybenzamine or pharmaceutically acceptable derivatives thereof, provided that each of the antibiotic resistance breakers is different.
  • the antibiotic resistance breakers used in the present invention have different mechanisms of action.
  • the term "mechanism of action” describes the specific biochemical interaction through which the ARB produces its pharmacological effect. In other words, how the ARB restores the efficacy (i.e. antibacterial activity) of the antibacterial agent against the bacterium of interest.
  • the mechanism of actions of the ARBs mefloquine, suloctidil, zidovudine and phenoxybenzamine are discussed in more detail in the Examples.
  • Mechanism of action and mode of action are used interchangeably; typically referring to the way in which the drug interacts and produces a medical effect.
  • a mode of action describes functional or anatomical changes, at the cellular level, resulting from the exposure of a living organism to a substance. This differs from a mechanism of action, as it is a more specific term that focuses on the interaction between the drug itself and an enzyme or receptor and its particular form of interaction, whether through inhibition, activation, agonism, or antagonism.
  • antibiotic resistance breakers may work by disrupting the bacterial cell membrane, by destroying the bacterial cell wall, inhibiting resistance mechanisms such as drug efflux from the cell, inhibiting bacterial enzymes which destroy antibiotics, including enhancing access of antibiotics to drug targets, and preventing the direct modification or inactivation of antibiotics.
  • the present invention advantageously allows the lifetime of an antibacterial agent to be extended, despite the inevitable build-up of antibiotic resistance against the antibacterial agent alone and in combination with the antibiotic resistance breakers.
  • the antibacterial agent effectively becomes renewable.
  • the term "combining" or “in combination with” covers both separate and sequential use of the antibacterial agent and each antibiotic resistance breaker.
  • either the antibacterial agent or the antibiotic resistance breaker may be administered first.
  • administration is simultaneous, the compounds may be administered either in the same or a different pharmaceutical composition.
  • the time periods of the present invention will depend on the rate at which antibiotic resistance develops to the antibacterial agent, used alone or in combination with an antibiotic resistance breaker.
  • the time periods can for instance be identical or different in length, for example, the first time period could be longer, shorter or the same length as the second time period.
  • the second time period could be longer, shorter or the same length as the third time period, etc.
  • each of the time periods is between 5 and 50 years, preferably between 10 and 40 years, more preferably between 20 and 30 years.
  • antibacterial agent refers to compounds which kill bacteria or inhibit their growth.
  • Antibacterial agents are used in the treatment and prevention of bacterial infections and are commonly classified based on their mechanism of action, chemical structure or spectrum of activity. For example, penicillins and cephalosporins target the bacterial cell wall, polymyxins target the cell membrane, rifamycins, lipiarmycins, quinolones and sulphonamides interfere with essential bacterial enzymes, and macrolides, lincosamides and tetracyclines target protein synthesis.
  • Suitable antibacterial agents for the present invention include one or more compounds selected from the following:
  • ⁇ -Lactams including:
  • penicillins such as (I) benzylpenicillin, procaine benzylpenicillin, phenoxy-methylpenicillin, methicillin, propicillin, epicillin, cyclacillin, hetacillin, 6-aminopenicillanic acid, penicillic acid, penicillanic acid sulphone (sulbactam), penicillin G, penicillin V, phenethicillin, phenoxymethylpenicillinic acid, azlocillin, carbenicillin, cloxacillin, D-(-)-penicillamine, dicloxacillin, nafcillin and oxacillin,
  • penicillins such as (I) benzylpenicillin, procaine benzylpenicillin, phenoxy-methylpenicillin, methicillin, propicillin, epicillin, cyclacillin, hetacillin, 6-aminopenicillanic acid, penicillic acid, penicillanic acid sulphone (sul
  • penicillinase-resistant penicillins e.g. flucloxacillin
  • antipseudomonal penicillins e.g. carboxypenicillins such as ticarcillin or ureidopenicillins such as piperacillin
  • carboxypenicillins such as ticarcillin or ureidopenicillins such as piperacillin
  • cephalosporins such as cefaclor, cefadroxil, cefalexin (cephalexin), cefcapene, cefcapene pivoxil, cefdinir, cefditoren, cefditoren pivoxil, cefixime, cefotaxime, cefpirome, cefpodoxime, cefpodoxime proxetil, cefprozil, cefradine, ceftazidime, cefteram, cefteram pivoxil, ceftriaxone, cefuroxime, cefuroxime axetil, cephaloridine, cephacetrile, cephamandole, cephaloglycine, ceftobiprole, PPI-0903 (TAK-599), 7-aminocephalosporanic acid, 7-aminodes- acetoxycephalosporanic acid, cefamandole, cefazolin, cefmetazole, ce
  • ⁇ -lactams such as monobactams (e.g. aztreonam), carbapenems (e.g. imipenem (optionally in combination with a renal enzyme inhibitor such as cilastatin), meropenem, ertapenem, doripenem (S-4661) and RO4908463 (CS-023)), penems (e.g. faropenem) and 1-oxa ⁇ -lactams (e.g. moxalactam).
  • monobactams e.g. aztreonam
  • carbapenems e.g. imipenem (optionally in combination with a renal enzyme inhibitor such as cilastatin), meropenem, ertapenem, doripenem (S-4661) and RO4908463 (CS-023)
  • penems e.g. faropenem
  • 1-oxa ⁇ -lactams e.g. moxalactam
  • Tetracyclines such as tetracycline, demeclocycline, doxycycline, lymecycline, minocycline, oxytetracycline, chlortetracycline, meclocycline and methacycline, as well as glycylcyclines (e.g. tigecycline).
  • Aminoglycosides such as amikacin, gentamicin, netilmicin, neomycin, streptomycin, tobramycin, amastatin, butirosin, butirosin A, daunorubicin, dibekacin, dihydrostreptomycin, G 418, hygromycin B, kanamycin B, kanamycin, kirromycin, paromomycin, ribostamycin, sisomicin, spectinomycin, streptozocin and thiostrepton.
  • Aminoglycosides such as amikacin, gentamicin, netilmicin, neomycin, streptomycin, tobramycin, amastatin, butirosin, butirosin A, daunorubicin, dibekacin, dihydrostreptomycin, G 418, hygromycin B, kanamycin B, kanamycin, kirromycin, paromomycin
  • Macro I ides such as azithromycin, clarithromycin, erythromycin, roxithromycin, spiramycin, amphotericins B (e.g. amphotericin B), bafilomycins (e.g. bafilomycin A1), brefeldins (e.g. brefeldin A), concanamycins (e.g. concanamycin A), filipin complex, josamycin, mepartricin, midecamycin, nonactin, nystatin, oleandomycin, oligomycins (e.g. oligomycin A, oligomycin
  • Ketolides such as telithromycin and cethromycin (ABT-773).
  • Lincosamines such as lincomycin.
  • Phenicols such as chloramphenicol and thiamphenicol.
  • Steroids such as fusidic acid (optionally in metal salt form, e.g. in salt form with an alkali metal such as sodium).
  • Glycopeptides such as vancomycin, teicoplanin, bleomycin, phleomycin, ristomycin, telavancin, dalbavancin and oritavancin.
  • Streptogramins such as quinupristin and dalfopristin, or a combination thereof.
  • Peptides such as polymyxins (e.g. polymyxin E (colistin) and polymyxin B), lysostaphin, duramycin, actinomycins (e.g. actinomycin C and actinomycin D), actinonin, 7-aminoactinomycin D, antimycin A, antipain, bacitracin, cyclosporin
  • echinomycin echinomycin
  • gramicidins e.g. gramicidin A and gramicidin C
  • myxothiazol nisin
  • paracelsin valinomycin
  • viomycin echinomycin
  • Sulfonamides such as sulfamethoxazole, sulfadiazine, sulfaquinoxaline, sulfathiazole (which latter two agents are optionally in metal salt form, e.g. in salt form with an alkali metal such as sodium), succinylsulfathiazole, sulfadimethoxine, sulfaguanidine, sulfamethazine, sulfamonomethoxine, sulfanilamide and sulfasalazine.
  • sulfamethoxazole sulfadiazine
  • sulfaquinoxaline e.g. in salt form with an alkali metal such as sodium
  • succinylsulfathiazole sulfadimethoxine
  • sulfaguanidine sulfamethazine
  • sulfamonomethoxine s
  • Trimethoprim optionally in combination with a sulfonamide, such as sulfamethoxazole (e.g. the combination co-trimoxazole).
  • a sulfonamide such as sulfamethoxazole (e.g. the combination co-trimoxazole).
  • Antituberculous drugs such as isoniazid, rifampicin, rifabutin, pyrazinamide, ethambutol, streptomycin, amikacin, capreomycin, kanamycin, quinolones (e.g. those at (q) below), para-aminosalicylic acid, cycloserine and ethionamide.
  • Antileprotic drugs such as dapsone, rifampicin and clofazimine.
  • Nitrofurans such as nitrofurantoin.
  • Quinolones such as nalidixic acid, norfloxacin, ciprofloxacin, ofloxacin, levofloxacin, moxifloxacin, gatifloxacin, gemifloxacin, garenoxacin, DX-619, WCK 771 (the arginine salt of S-(-)-nadifloxacin), 8-quinolinol, cinoxacin, enrofloxacin, flumequine, lomefloxacin, oxolinic acid and pipemidic acid.
  • Amino acid derivatives such as azaserine, bestatin, D-cycloserine, 1 ,10- phenanthroline, 6-diazo-5-oxo-L-norleucine and L-alanyl-L-1-aminoethyl-phosphonic acid.
  • Aureolic acids such as chromomycin A3, mithramycin A and mitomycin C C.
  • Glucosamines such as 1-deoxymannojirimycin, 1-deoxynojirimycin and A/-methyl-1- deoxynojirimycin.
  • Diaminopyrimidines such as iclaprim (AR-100).
  • Taxoids such as paclitaxel.
  • Statins such as mevastatin.
  • Polyethers such as lasalocid A, lonomycin A, monensin, nigericin and salinomycin.
  • Peptidyl nucleosides such as blasticidine S, nikkomycin, nourseothricin and puromycin.
  • Nucleosides such as adenine 9 ⁇ -D-arabinofuranoside, 5-azacytidine, cordycepin, formycin A, tubercidin and tunicamycin.
  • Pleuromutilins such as GSK-565154, GSK-275833 and tiamulin.
  • Peptide deformylase inhibitors such as LBM415 (NVP PDF-713) and BB 83698.
  • Antibacterial agents for the skin such as fucidin, benzamycin, clindamycin, erythromycin, tetracycline, silver sulfadiazine, chlortetracycline, metronidazole, mupirocin, framycitin, gramicidin, neomycin sulfate, polymyxins (e.g. polymyxin B or polymyxin E) and gentamycin.
  • Miscellaneous agents such as methenamine (hexamine), doxorubicin, piericidin A, stigmatellin, actidione, anisomycin, apramycin, coumermycin A1 , L(+)-lactic acid, cytochalasins (e.g. cytochalasin B and cytochalasin D), emetine and ionomycin.
  • Antiseptic agents such as chlorhexidine, phenol derivatives (e.g. thymol and triclosan), quarternary ammonium compounds (e.g.
  • benzalkonium chloride cetylpyridinium chloride, benzethonium chloride, cetrimonium bromide, cetrimonium chloride and cetrimonium stearate), octenidine dihydrochloride, and terpenes (e.g. terpinen-4-ol). or a pharmaceutically acceptable derivative thereof.
  • the antibacterial agent is a polymyxin selected from colistin (polymyxin E), polymyxin B, or pharmaceutically acceptable derivatives thereof. More preferably the antibacterial agent is colistin or a pharmaceutically acceptable derivative thereof.
  • the antibacterial agent is a polymyxin derivative of formula (I):
  • R1 is Dab
  • R2 is Thr
  • R3 is DThr
  • R4 is Dab
  • R5 is Dab
  • R6 is DPhe
  • R7 is Leu
  • R8 is Abu
  • R9 is Dab
  • R10 is Thr
  • R(FA) is octanoyl
  • R1 is absent
  • R2 is Thr
  • R3 is DSer
  • R4 is Dab
  • R5 is Dab
  • R6 is DPhe
  • R7 is Leu
  • R8 is Dab
  • R9 is Dab
  • R10 is Thr; and R(FA) is octanoyl;
  • the polymyxin derivatives of formula (I) are known compounds developed by Northern Antibiotics Oy.
  • Example 1 of WO20161 13470 (A1) discloses the synthesis of NAB815.
  • the disclosure of this reference insofar as it relates to NAB815 is incorporated herein by reference.
  • polymyxin derivative of formula (I) is NAB 815, wherein R1 is Dab; R2 is Thr; R3 is DThr; R4 is Dab; R5 is Dab; R6 is DPhe; R7 is Leu; R8 is Abu; R9 is Dab; R10 is Thr; and R(FA) is octanoyl.
  • polymyxin derivative of formula (I) has R1-R10 representing an amino acid sequence Thr-DSer-cy[Dab-Dab-DPhe-Leu-Dab-Dab-Thr-]. In one embodiment the polymyxin derivative of formula (I) has R1-R10 representing SEQ ID NO. 2 defined herein.
  • SEQ ID NO.2 is Thr-DSer-cy[Dab-Dab-DPhe-Leu-Dab-Dab-Thr-].
  • SEQ ID NO.2 can also be written as Thr-DSer-[cyclo-Dbu-Dbu-DPhe-Leu-Dbu-Dbu-Thr] where Thr refers to Threonine; DSer refers to D-Serine; Dbu refers to 2,4-diaminobutyric acid; DPhe refers to D- Phenylalanine; Leu refers to Leucine and residues 3-9 form the cyclic heptapeptide portion.
  • polymyxin derivative of formula (I) has R1-R10 representing the amino acid sequence Dab-Thr-DThr-cy[Dab-Dab-DPhe-Leu-Abu-Dab-Thr-].
  • polymyxin derivative of formula (I) has R1-R10 representing SEQ ID NO. 1 defined herein.
  • polymyxin derivative of formula (I) has R1-R10 representing SEQ ID N0.1 defined herein.
  • SEQ ID N0.1 is Dab-Thr-DThr-cy[Dab-Dab-DPhe-Leu-Abu-Dab-Thr-].
  • SEQ ID N0.1 can also be written as Dbu-Thr-DThr-[cyclo -Dbu-Dbu-DPhe-Leu-Abu-Dbu-Thr] where Thr refers to Threonine; Dbu refers to 2,4-diaminobutyric acid; DThr refers to D- Threonine; DPhe refers to D-Phenylalanine; Leu refers to Leucine; Abu refers to 2- aminobutyric acid; and residues 4-10 form the cyclic heptapeptide portion.
  • the polymyxin derivative of formula (I) can also be represented as OA-Thr-DSer-cy[Dab- Dab-DPhe-Leu-Dab-Dab-Thr-], i.e. OA-SEQ ID NO. 2 or as OA-Dab-Thr-DThr-cy[Dab-Dab- DPhe-Leu-Abu-Dab-Thr-], i.e. OA-SEQ ID NO. 1.
  • OA-SEQ ID NO. 1 is preferred.
  • Both NAB 815 and NAB 739 have a total number of positive charges at physiological pH of 3.
  • these positive charges are located at positions R5, R8 and R9 of formula (I), i.e. within the heptapeptide ring.
  • NAB 815 two of these positive charges are located within the heptapeptide ring at positions R5 and R9, and one is located on the hydrophobic tail at position R1.
  • physiological pH refers to a pH value of more than 7.0 and below 7.6, such as a pH value in the range of from 7.1 to 7.5, for example in the range of from 7.2 to 7.4.
  • the antibacterial agent is a compound capable of killing log-phase or fast growing bacteria.
  • the antibacterial agent is a compound capable of killing clinically latent microorganisms, preferably a compound capable of killing clinically latent bacteria.
  • kill means a loss of viability as assessed by a lack of metabolic activity and "clinically latent microorganism” means a microorganism that is metabolically active but has a growth rate that is below the threshold of infectious disease expression.
  • the threshold of infectious disease expression refers to the growth rate threshold below which symptoms of infectious disease in a host are absent.
  • the metabolic activity of clinically latent microorganisms can be determined by several methods known to those skilled in the art; for example, by measuring mRNA levels in the microorganisms or by determining their rate of uridine uptake.
  • clinically latent microorganisms when compared to microorganisms under logarithmic growth conditions (in vitro or in vivo), possess reduced but still significant levels of:
  • mRNA e.g. from 0.0001 to 50%, such as from 1 to 30, 5 to 25 or 10 to 20%, of the level of mRNA
  • uridine e.g. [ 3 H]uridine
  • uptake e.g. from 0.0005 to 50%, such as from 1 to 40, 15 to 35 or 20 to 30% of the level of [ 3 H]uridine uptake.
  • Clinically latent microorganisms typically possess a number of identifiable characteristics. For example, they may be viable but non-culturable; i.e. they cannot typically be detected by standard culture techniques, but are detectable and quantifiable by techniques such as broth dilution counting, microscopy, or molecular techniques such as polymerase chain reaction.
  • clinically latent microorganisms are phenotypically tolerant, and as such are sensitive (in log phase) to the biostatic effects of conventional antibacterial agents (i.e.
  • microorganisms for which the minimum inhibitory concentration (MIC) of a conventional antibacterial is substantially unchanged); but possess drastically decreased susceptibility to drug-induced killing e.g. microorganisms for which, with any given conventional antibacterial agent, the ratio of minimum microbiocidal concentration (e.g. minimum bactericidal concentration, MBC) to MIC is 10 or more).
  • Test procedures that may be employed to determine the biological (e.g. bactericidal or antibacterial) activity of the active ingredients include those known to persons skilled in the art for determining:
  • methods for determining activity against clinically latent bacteria include a determination, under conditions known to those skilled in the art (such as those described in Nature Reviews, Drug Discovery 1 , 895-910 (2002), the disclosures of which are hereby incorporated by reference), of Minimum Stationary-cidal Concentration (“MSC”) or Minimum Dormicidal Concentration (“MDC”) for a test compound.
  • MSC Minimum Stationary-cidal Concentration
  • MDC Minimum Dormicidal Concentration
  • WO2000028074 describes a suitable method of screening compounds to determine their ability to kill clinically latent microorganisms.
  • a typical method may include the following steps:
  • the phenotypically resistant sub-population may be seen as representative of clinically latent bacteria which remain metabolically active in vivo and which can result in relapse or onset of disease.
  • methods for determining activity against log phase bacteria include a determination, under standard conditions (i.e. conditions known to those skilled in the art, such as those described in WO 2005014585, the disclosures of which document are hereby incorporated by reference), of MIC or MBC for a test compound.
  • the term "microorganisms” means fungi and bacteria.
  • microorganisms means bacteria.
  • Examples of compounds capable of killing clinically latent bacteria include those compounds disclosed in International Patent Application, Publication Numbers WO2007054693, WO2008117079 and WO2008142384. These applications describe suitable methods for the preparation of such compounds and doses for their administration.
  • an antibacterial agent include a compound selected from the group consisting of:
  • antibacterial agent include a compound selected from the group consisting of:
  • antibacterial agent include a compound selected from the group consisting of:
  • furan-2-carboxylic acid [2-methyl-4-(3-phenylpyrrolidin-1 -yl)quinolin-6-yl]amide
  • antibacterial agents are 4-methyl-1 -(2-phenylethyl)-8-phenoxy-2,3- dihydro-1 H-pyrrolo[3,2-c]-quinoline (Example 9, WO2007054693), 4-(3-benzylpyrrolidin-1 -yl)- 2-methyl-6-phenoxyquinoline (Example 8, WO2008142384), and N-[4-(3-benzylpyrrolidin-1 - yl)-2-methylquinolin-6-yl]benzamide (Example 38, WO2008142384), and pharmaceutically acceptable derivatives thereof.
  • the antibacterial agent is 4-(3-benzylpyrrolidin-1-yl)-2- methyl-6-phenoxyquinoline, N-[4-(3-benzylpyrrolidin-1 -yl)-2-methylquinolin-6-yl]benzamide or a pharmaceutically acceptable derivative thereof.
  • a more preferred antibacterial agent is 4- methyl-1-(2-phenylethyl)-8-phenoxy-2,3-dihydro-1 H-pyrrolo[3,2-c]-quinoline or a pharmaceutically acceptable derivative thereof such as the hydrochloride salt thereof.
  • bacteria includes, but is not limited to, references to organisms (or infections due to organisms) of the following classes and specific types:
  • Gram-positive cocci such as Staphylococci (e.g. Staph, aureus, Staph, epidermidis, Staph. saprophyticus, Staph, auricularis, Staph, capitis capitis, Staph, c. ureolyticus, Staph, caprae,
  • Staphylococci e.g. Staph, aureus, Staph, epidermidis, Staph. saprophyticus, Staph, auricularis, Staph, capitis capitis, Staph, c. ureolyticus, Staph, caprae
  • Streptococci e.g.beta-haemolytic, pyogenic streptococci (such as Strept. agalactiae, Strept. canis, Strept. dysgalactiae dysgalactiae, Strept. dysgalactiae equisimilis,
  • Strept. equi equi Strept. equi zooepidemicus, Strept. iniae, Strept. porcinus and Strept. pyogenes
  • microaerophilic, pyogenic streptococci Streptococcus "milleri", such as Strept. anginosus, Strept. constellatus constellatus, Strept. constellatus pharyngidis and Strept. intermedius
  • oral streptococci of the "mitis” alpha-haemolytic - Streptococcus "viridans", such as Strept. mitis, Strept. oralis, Strept. sanguinis, Strept. cristatus, Strept. gordonii and
  • Strept. parasanguinis "salivarius” (non-haemolytic, such as Strept. salivarius and Strept. vestibularis) and "mutans” (tooth-surface streptococci, such as Strept. criceti, Strept. mutans,
  • Strept. ratti and Strept. sobrinus groups, Strept. acidominimus, Strept. bovis, Strept. faecalis, Strept. equinus, Strept. pneumoniae and Strept. suis, or Streptococci alternatively classified as Group A, B, C, D, E, G, L, P, U or V Streptococcus);
  • Gram-negative cocci such as Neisseria gonorrhoeae, Neisseria meningitidis, Neisseria cinerea, Neisseria elongata, Neisseria flavescens, Neisseria lactamica, Neisseria mucosa, Neisseria sicca, Neisseria subflava and Neisseria weaveri; Bacillaceae, such as Bacillus anthracis, Bacillus subtilis, Bacillus thuringiensis, Bacillus stearothermophilus and Bacillus cereus; Enterobacteriaceae, such as Escherichia coli, Enterobacter (e.g.
  • Enterobacter aerogenes Enterobacter aerogenes, Enterobacter agglomerans and Enterobacter cloacae
  • Citrobacter such as Citrob. freundii and Citrob. divernis
  • Hafnia e.g. Hafnia alvei
  • Erwinia e.g. Erwinia persicinus
  • Morganella morganii Salmonella (Salmonella enterica and Salmonella typhi), Shigella (e.g. Shigella dysenteriae, Shigella flexneri, Shigella boydii and Shigella sonnei), Klebsiella (e.g. Klebs. pneumoniae, Klebs. oxytoca, Klebs.
  • Shigella dysenteriae Shigella flexneri
  • Klebsiella e.g. Klebs. pneumoniae, Klebs. oxytoca, Klebs.
  • Enterococci e.g. Enterococcus avium, Enterococcus casseliflavus, Enterococcus cecorum, Enterococcus dispar, Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Enterococcus flavescens, Enterococcus gallinarum, Enterococcus hirae, Enterococcus malodoratus, Enterococcus mundtii, Enterococcus pseudoavium, Enterococcus raffinosus and Enterococcus solitarius); Helicobacter (e.g.
  • Helicobacter pylori Helicobacter cinaedi and Helicobacter fennelliae
  • Acinetobacter e.g. A. baumanii, A. calcoaceticus, A. haemolyticus, A. johnsonii, A. junii, A. Iwoffi and A. radioresistens
  • Pseudomonas e.g. Ps. aeruginosa, Ps. maltophilia (Stenotrophomonas maltophilia), Ps. alcaligenes, Ps. chlororaphis, Ps. fluorescens, Ps. luteola. Ps. mendocina, Ps. monteilii, Ps.
  • oryzihabitans Ps. pertocinogena, Ps. pseudalcaligenes, Ps. putida and Ps. stutzeri); Bacteriodes fragilis; Peptococcus (e.g. Peptococcus niger); Peptostreptococcus; Clostridium (e.g. C. perfringens, C. difficile, C. botulinum, C. tetani, C. absonum, C. argentinense, C. baratii, C. bifermentans, C. beijerinckii, C. butyricum, C. cadaveris, C. carnis, C. celatum, C.
  • Clostridium e.g. C. perfringens, C. difficile, C. botulinum, C. tetani, C. absonum, C. argentinense, C. baratii, C. bifermentans, C. beijerinckii, C
  • clostridioforme C. cochlearium, C. cocleatum, C. fallax, C. ghonii, C. glycolicum, C. haemolyticum, C. hastiforme, C. histolyticum, C. indolis, C. innocuum, C. irregulare, C. leptum, C. limosum, C. malenominatum, C. novyi, C. oroticum, C. paraputrificum, C. piliforme, C. putrefasciens, C. ramosum, C. septicum, C. sordelii, C. sphenoides, C. sporogenes, C.
  • Mycoplasma e.g. M. pneumoniae, M. hominis, M. genitalium and M. urealyticum
  • Mycobacteria e.g.
  • Mycobacterium tuberculosis Mycobacterium avium, Mycobacterium fortuitum, Mycobacterium marinum, Mycobacterium kansasii, Mycobacterium chelonae, Mycobacterium abscessus, Mycobacterium leprae, Mycobacterium smegmitis, Mycobacterium africanum, Mycobacterium alvei, Mycobacterium asiaticum, Mycobacterium aurum, Mycobacterium bohemicum, Mycobacterium bovis, Mycobacterium branderi, Mycobacterium brumae, Mycobacterium celatum, Mycobacterium chubense, Mycobacterium confluentis, Mycobacterium conspicuum, Mycobacterium cookii, Mycobacterium flavescens, Mycobacterium gadium, Mycobacterium gastri, Mycobacterium genavense, Mycobacterium gordonae, Mycobacterium goodii, Mycobacterium haemophilum
  • Brucella abortus Brucella canis, Brucella melintensis and Brucella suis
  • Campylobacter e.g. Campylobacter jejuni, Campylobacter coli, Campylobacter lari and Campylobacter fetus
  • Listeria monocytogenes Vibrio (e.g.
  • Vibrio cholerae and Vibrio parahaemolyticus Vibrio alginolyticus, Vibrio carchariae, Vibrio fluvialis, Vibrio furnissii, Vibrio hollisae, Vibrio metschnikovii, Vibrio mimicus and Vibrio vulnificus); Erysipelothrix rhusopathiae; Corynebacteriaceae (e.g. Corynebacterium diphtheriae, Corynebacterium jeikeum and Corynebacterium urealyticum); Spirochaetaceae, such as Borrelia (e.g.
  • Pasteurella e.g. Pasteurella aerogenes, Pasteurella bettyae, Pasteurella canis, Pasteurella dagmatis, Pasteurella gallinarum, Pasteurella haemolytica, Pasteurella multocida multocida, Pasteurella multocida gallicida, Pasteurella multocida septica, Pasteurella pneumotropica and Pasteurella stomatis
  • Bordetella e.g.
  • Nocardiaceae such as Nocardia (e.g. Nocardia asteroides and Nocardia brasiliensis); Rickettsia (e.g. Ricksettsii or Coxiella burnetii); Legionella (e.g.
  • Moraxella catarrhalis Moraxella catarrhalis; Cyclospora cayetanensis; Entamoeba histolytica; Giardia lamblia; Trichomonas vaginalis; Toxoplasma gondii; Stenotrophomonas maltophilia; Burkholderia ce
  • Capnocytophaga e.g. Capnocytophaga canimorsus, Capnocytophaga cynodegmi, Capnocytophaga gingivalis, Capnocytophaga granulosa, Capnocytophaga haemolytica, Capnocytophaga ochracea and Capnocytophaga sputtona
  • Bartonella ⁇ Bartonella bacilliformis Bartonella clarridgeiae, Bartonella elizabethae, Bartonella henselae, Bartonella quintana and Bartonella vinsonii arupensis
  • Leptospira e.g.
  • Fusobacterium e.g. F. gonadiaformans, F. mortiferum, F. naviforme, F. necrogenes, F. necrophorum necrophorum, F.
  • Chlamydia e.g. Chlamydia trachomatis
  • Cryptosporidium e.g. C. parvum, C. hominis, C. canis, C. felis, C. meleagridis and C. muris
  • Chlamydophila e.g.
  • Leuconostoc e.g. Leuconostoc citreum, Leuconostoc cremoris, Leuconostoc d
  • the bacteria are Gram-negative bacteria, e.g. Enterobacteriaceae, such as Escherichia coli, Klebsiella (e.g. Klebs. pneumoniae and Klebs. oxytoca) and Proteus (e.g. Pr. mirabilis, Pr. rettgeri and Pr. vulgaris); Haemophilis influenzae; Mycobacteria, such as Mycobacterium tuberculosis; and Enterobacter (e.g. Enterobacter cloacae).
  • the bacteria are Enterobacteriaceae, such as Escherichia coli and Klebsiella (e.g. Klebs. pneumoniae and Klebs. oxytoca).
  • Acinetobacter spp Particularly preferred are Escherichia coli, and Klebs. pneumoniae (e.g. Klebs. pneumoniae subsp. pneumoniae). Also particularly preferred are Acinetobacter spp. (e.g. Acinetobacter baumannii).
  • Suitable acid addition salts include carboxylate salts (e.g. formate, acetate, trifluoroacetate, propionate, isobutyrate, heptanoate, decanoate, caprate, caprylate, stearate, acrylate, caproate, propiolate, ascorbate, citrate, glucuronate, glutamate, glycolate, a-hydroxybutyrate, lactate, tartrate, phenylacetate, mandelate, phenylpropionate, phenylbutyrate, benzoate, chlorobenzoate, methylbenzoate, hydroxy benzoate, methoxybenzoate, dinitrobenzoate, o- acetoxybenzoate, salicylate, nicotinate, isonicotinate, cinnamate, oxalate, malonate, succinate, suberate, sebacate, fumarate, malate, maleate, hydroxymaleate, hippurate, phthalate or tere
  • sulfonate salts e.g. benzenesulfonate, methyl-, bromo- or chloro-benzenesulfonate, xylenesulfonate, methanesulfonate, ethanesulfonate, propanesulfonate, hydroxyethanesulfonate, 1- or 2- naphthalene-sulfonate or 1 ,5-naphthalenedisulfonate salts) or sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate or nitrate salts, and the like.
  • sulfonate salts e.g. benzenesulfonate, methyl-, bromo- or chloro-benzenesulfonate, xylenesulfonate, methanesulf
  • the compound may be colistin sulfate, colistimethate sodium, colistin sodium methanesulfonate, or polymyxin B sulfate. Particularly preferred is colistin, colistin sulfate, colistin sodium methane sulfonate or colistimethate sodium, e.g. colistin or colistimethate sodium.
  • a preferred salt of polymyxin B is the sulfate salt thereof, i.e. polymyxin B sulfate.
  • a preferred salt of polymyxin E is the sulfate salt thereof, i.e. polymyxin E sulfate.
  • a typically used acid for formation of the pharmaceutically acceptable salt of the polymyxin derivative of formula (I) is sulfuric acid.
  • the antibacterial agent for use in the present invention is commercially available or can be prepared by synthesis methods known in the art.
  • polymyxin B, polymyxin B sulfate, polymyxin E sulfate and polymyxin E are commercially available from Sigma Aldrich Limited or Finetech Industry Limited. Suitable dosages and formulations are known in the art.
  • at least one of the antibiotic resistance breakers for use in the present invention is preferably a known non-antibiotic drug.
  • Such drugs are typically commercially available or can be prepared by synthesis methods known in the art.
  • mefloquine, mefloquine hydrochloride, zidovudine, suloctidil and phenoxybenzamine hydrochloride are commercially available from Sigma Aldrich Limited. Suitable dosages and formulations of these drugs are also known in the art.
  • a preferred salt of mefloquine is the hydrochloride salt thereof, i.e. mefloquine hydrochloride.
  • Mefloquine is a chiral molecule with two asymmetric carbon centres, which means that it has four different stereoisomers. Mefloquine is commercially available as a racemate of the (R,S)- and (S,R)-enantiomers:
  • mefloquine includes all enantiomers, tautomers and stereoisomers thereof.
  • the corresponding enantiomers and/or tautomers may also be isolated and/or prepared by methods known in the art.
  • Suloctidil is preferably used in its non-salt form.
  • Phenoxybenzamine is preferably used as the hydrochloride salt.
  • Zidovudine is preferably used in its non-salt form.
  • the compounds used in the present invention may be as the raw materials, but are preferably provided in the form of pharmaceutical compositions or formulations.
  • the antibacterial agent and at least two antibiotic resistance breakers may be used either as separate formulations or as a single combined formulation. When combined in the same formulation it will be appreciated that the two compounds must be stable and compatible with each other and the other components of the formulation.
  • the formulation can be suitable for oral, parenteral (including subcutaneous e.g. by injection or by depot tablet, intradermal, intrathecal, intramuscular e.g. by depot and intravenous), rectal and topical (including dermal, buccal and sublingual) or in a form suitable for administration by inhalation or insufflation administration.
  • the compositions of the invention are formulated for systemic intravenous, intramuscular injection or inhaled, oral or topical administration. Examples
  • Example 1 Microbiological profiling (FIC assay) of suloctidil, phenoxybenzamine hydrochloride and mefloquine in combination with polymyxin B nonapeptide against E. co// " To I C and E. coli D22 mutant strains
  • FIC assay Microbiological profiling (FIC assay) of suloctidil, phenoxybenzamine hydrochloride and mefloquine in combination with polymyxin B nonapeptide against E. co// " To I C and E. coli D22 mutant strains
  • FIC assay Microbiological profiling
  • All compounds were obtained from a commercial source such as Sigma Aldrich.
  • the bacterial strains (wild-type and mutant) were obtained from commercially available bacterial collections such as ATCC.
  • Broth microdilution chequerboard assay was performed following the procedure described in the Clinical Microbiology Procedures Handbook (Isenberg, H.D. Ed.) American Society for Microbiology, Washington, DC. MIC assays were made using EUCAST & CLSI-approved guidelines using CM0473 ISO-SENSITESTTM BROTH medium.
  • the MIC for colistin and suloctidil were obtained against the wild- type strains: E. coli ATCC 25922 and E. coli ATCC BAA 2471.
  • the MIC is the lowest concentration of a drug that will inhibit visible growth of a microorganism after overnight incubation.
  • the MIC values (in mcg/ml) are shown in Table 1 below.
  • E. coli mutant strains To determine whether each of the ARBs suloctidil, phenoxybenzamine and mefloquine had the same mechanism of action when combined with colistin, each ARB was combined with polymyxin B nonapeptide and tested against E.coli mutant strains.
  • Polymyxin antibiotics are known to bind to lipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria and disrupt both the outer and inner membranes. They have a general structure consisting of a cyclic peptide with a hydrophobic tail and disrupt the structure of the bacterial cell membrane by interacting with its phospholipids. The hydrophobic tail is believed to be important in causing the membrane damage.
  • LPS lipopolysaccharide
  • polymyxin B nonapeptide Removal of the hydrophobic tail of polymyxin B produces polymyxin B nonapeptide. This compound still binds to LPS but no longer kills the bacterial cell. Additionally it still detectably increases the permeability of the bacterial cell wall to other compounds.
  • polymyxin B nonapeptide was therefore used instead of colistin.
  • the E. coli TolC mutant strain was derived from the wild E.coli strain ATCC 25922 but unlike the wild strain, it did not express the AcrAB-TolC multidrug efflux pump.
  • This pump assembly includes the outer-membrane channel TolC, the secondary transporter AcrB located in the inner membrane, and the periplasmic AcrA which bridges these two integral membrane proteins.
  • the AcrAB-TolC efflux pump is known to transport a wide variety of compounds out of the bacterial cell, and confer resistance to a broad spectrum of antibiotics.
  • the MIC values (in mcg/ml) for suloctidil, mefloquine, phenoxybenzamine hydrochloride and polymyxin B nonapeptide with this mutant strain are shown in Table 2 below, along with the MIC values with the wild E.coli strain ATCC 25922 from Table 1 above.
  • suloctidil and polymyxin B nonapepitde resulted in synergism with suloctidil at 2 mcg/ml in combination with polymyxin B nonapeptide at 0.5, 1 , 4, 16 or 32 mcg/ml, and with suloctidil at 4 mcg/ml in combination with polymyxin B nonapeptide at 0.25 mcg/ml.
  • the E.coli D22 mutant (lpxC101 proA23 lac-28 tsx-81 trp-30 his-51 rpsL173(strR) tufA1 ampCp-1) was derived from the wild E.coli strain ATCC 25922 but unlike the wild strain had a reduced biosynthesis of lipid A.
  • Lipid A is a lipid component held responsible for the toxicity of gram-negative bacteria. It is the innermost of the three regions of the lipopolysaccharide (LPS), and its hydrophobic nature allows it to anchor the LPS to the outer membrane.
  • suloctidil and polymyxin B nonapeptide resulted in synergism with suloctidil at 2 mcg/ml in combination with polymyxin B nonapeptide at 2, 4, 8, 16 or 32 mcg/ml, and with suloctidil at 4 mcg/ml in combination with polymyxin B nonapeptide at 0.25 or 0.5 mcg/ml.
  • the compounds were obtained from a commercial source, e.g. Sigma Aldrich.
  • the bacterial strains were obtained from a commercially available bacterial collection such as ATCC.
  • Log phase growth of the bacterial strains was carried out as known in the art.
  • the log phase bacterial cultures were then incubated with each of the combinations overnight by diluting with nutrient broth (Oxoid) to 10 7 CFU/ml and adding 280 ⁇ of each culture to each well to make a final concentration of 300 ⁇ . Incubation of the combinations with the bacterial suspensions was carried out for 24 hours.
  • CS colistin sulfate
  • PBNH polymyxin B nonapeptide hydrochloride
  • S suloctidil
  • M mefloquine
  • POBH phenoxybenzamine hydrochloride
  • BAA2469 and BAA2472 were susceptible to colistin sulfate with MIC at 0.031 mg/L for BAA2469 and 0.5 -1 mg/L for BAA2472.
  • Example 1 and Example 2 show three different patterns of response. This is important because it suggests different mechanisms of action for suloctidil, phenoxybenzamine hydrochloride and mefloquine.
  • the patterns can be summarized as follows:
  • Example 1 against the E. coli TolC mutant From the results in Example 1 against the E. coli TolC mutant, it can be seen that the combination with suloctidil has a synergistic effect and that suloctidil may have some antibiotic activity. This compound is also efflux-pump dependent and has ARB activity. From the results in Example 1 against the E. coli D22 mutant, it can be seen that the combination with suloctidil is synergistic and that LPS may be a barrier.
  • suloctidil as an ARB is not dependent on permeability of the bacterial cell wall, is dependent on efflux and LPS, and may have its own antibiotic activity.
  • Example 2 It can be seen from the table in Example 2 that there was no interaction between colistin or polymyxin B hydrochloride nonapeptide with phenoxybenzamine against strain BAA2472. From the results in Example 1 against the E. coli TolC mutant, it can be seen that the combination with phenoxybenzamine hydrochloride did not have a synergistic effect. Against the E. coli D22 mutant, the combination with phenoxybenzamine hydrochloride was, however, synergistic.
  • Example 2 It can be seen from the table in Example 2 that the FICs for colistin and polymyxin B hydrochloride nonapeptide with mefloquine were similar. This suggests a role for increased permeabilization. From the results in Example 1 against the E. coli TolC mutant and the E. coli D22 mutant, it can be seen that the combinations with mefloquine were synergistic. Overall therefore mefloquine has a different synergy profile to suloctidil and phenoxybenzamine hydrochloride and consequently must have a different mechanism of action with the polymyxin antibacterial agent.
  • antibiotic resistance breakers (suloctidil, mefloquine and phenoxybenzamine hydrochloride) have a different mechanism of action with a polymyxin antibacterial agent. They are therefore suitable for use in the present invention to restore efficacy/prolong the lifetime of the antibacterial agent.
  • Samples of bacteria were grown in Lysogeny Broth at 30°C to OD 6 oo ⁇ 0.1. These cultures were then split and each sample treated with the appropriate concentration of the test compound. Samples were collected at two time points: 2 hours and 4 hours. 100 ⁇ _ of each sample was stained with 0.4 ⁇ SYTOX Green, 1.5 ⁇ .
  • DAPI 4,-6-diamidino-2- phenylindole; a blue-fluorescent DNA stain
  • FM 4-64 N-(3- triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl) pyridinium dibromide; a red-fluorescent dye.
  • Cells were centrifuged, re-suspended and transferred to an agarose pad (20% LB and 1 % agarose) for imaging. Membranes are shown in red, DNA in blue, and green staining indicates permeabilization of the cell membrane.
  • White scale bar is 1 ⁇ . The greyscale copy of the images is included as the B version of the relevant Figure.
  • the characteristic phenotype produced by treatment with the indicated compounds for 2 hours is shown in Figure 2 (2A and 2B).
  • the effect of each of the compounds when used alone can be determined by comparing the control (DMSO 1.28%) with the image for the compound in question (e.g. colistin 2.5).
  • FIG. 5 The predominant phenotype produced by treatment of E.coli ATCC 25922 with colistin and/or suloctidil for 2 hours is shown in Figure 5 (5A and 5B).
  • Example 4 Additional Mechanism action studies with zidovudine (azidothymidine) and control compounds
  • additional studies were carried out to obtain more information on the mechanism by which zidovudine (azidothymidine) targets DNA replication.
  • These studies involved first looking at the effect of zidovudine and other control compounds on E.coli tolC, and second comparing zidovudine to other DNA replication inhibitors.
  • the compounds, their class and their target are shown below in Table 9: Table 9
  • BCP Bacterial Cytological Profiling
  • Figures 1 1 to 18 BCP images for the control, zidovudine and a compound of each class of antibiotic tested are contained in Figures 1 1 to 18.
  • Figure 11 (1 1 A and 11 B) includes the images obtained for the DMSO control: E.coli tolC treated with 0.5% DMSO for 30 minutes and 120 minutes.
  • Figure 12 (12A and 12B) includes the images obtained for E.coli tolC treated with 0.06 ⁇ g/mL zidovudine for 30 minutes and 120 minutes.
  • Figure 13 (13A and 13B) includes the images obtained for E.coli tolC treated with 0.3 ⁇ g/mL zidovudine (azidothymidine) for 30 minutes and 120 minutes.
  • Figure 14 includes the images obtained for E.coli tolC treated with 0.002 ⁇ g/mL ciprofloxacin for 30 minutes and 120 minutes.
  • Figure 15 includes the images obtained for E.coli tolC treated with 0.01 ⁇ g/mL ciprofloxacin for 30 minutes and 120 minutes.
  • Figure 16 (16A and 16B) includes the images obtained for E.coli tolC treated with 20 ⁇ g/mL cephalexin for 30 minutes and 120 minutes.
  • Figure 17 (17A and 17B) includes the images obtained for E.coli tolC treated with 25 ⁇ g/mL cerulenin for 120 minutes and 240 minutes.
  • Figure 18 (18A and 18B) includes the images obtained for E.coli tolC treated with 7.5 ⁇ g/mL rifampicin for 30 minutes and 120 minutes.
  • Figure 19 contains a clustergram of zidovudine (azidothymidine) and the different classes of antibiotic: transcription (rifampicin), lipid biosynthesis (cerulenin), cell wall biosynthesis (cephalexin), DNA replication (ciprofloxacin) with E.coli tolC treated at 5X MIC for 120 minutes (or for 240 minutes for cerulenin).
  • transcription rifampicin
  • lipid biosynthesis lipid biosynthesis
  • cephalexin cell wall biosynthesis
  • DNA replication ciprofloxacin
  • Figure 20 (20A and 20B) includes the images obtained for E.coli tolC treated with 1.5 ⁇ g/mL daunorubicin for 30 minutes and 120 minutes.
  • Figure 21 (21A and 21 B) includes the images obtained for E.coli tolC treated with 7.5 ⁇ g/mL daunorubicin for 30 minutes and 120 minutes.
  • Figure 22 (22A and 22B) includes the images obtained for E.coli tolC treated with 1 ⁇ g/mL novobiocin for 30 minutes and 120 minutes.
  • Figure 23 (23A and 23B) includes the images obtained for E.coli tolC treated with 5 ⁇ g/mL novobiocin for 30 minutes and 120 minutes.
  • Figure 24 includes the images obtained for E.coli tolC treated with 0.03 ⁇ g/mL mitomycin C for 30 minutes and 120 minutes.
  • Figure 25 includes the images obtained for E.coli tolC treated with 0.15 ⁇ g/mL mitomycin C for 30 minutes and 120 minutes.
  • Figure 26 contains a clustergram of zidovudine (azidothymidine) and the different types of DNA replication inhibitor: crosslinks DNA (mitomycin C), DNA gyrase A (ciprofloxacin), DNA intercalator (daunorubicin) and DNA gyrase B (novobiocin) with E.coli tolC treated at 5X MIC for 120 minutes.
  • zidovudine asthymidine
  • ciprofloxacin DNA intercalator
  • DNA gyrase B novobiocin

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Abstract

L'invention concerne un procédé pour restaurer l'efficacité d'un agent antibactérien, le procédé comprenant (i) la combinaison de l'agent antibactérien avec un premier agent de rupture de résistance aux antibiotiques après une première période de temps ; et (ii) la combinaison de l'agent antibactérien avec un second agent de rupture de résistance aux antibiotiques après une seconde période de temps, pendant chaque période de temps, l'efficacité de l'agent antibactérien diminue en raison de l'augmentation de la résistance aux antibiotiques, et chaque agent de rupture de résistance aux antibiotiques restaure au moins partiellement l'efficacité de l'agent antibactérien par rapport à la fin de la période de temps précédente. Les premier et second agents de rupture de résistance aux antibiotiques ont différents mécanismes d'action. L'invention concerne également l'utilisation d'au moins deux agents de rupture de résistance aux antibiotiques pour prolonger l'efficacité d'un agent antibactérien, lesdits au moins deux agents de rupture de résistance aux antibiotiques ayant différents mécanismes d'action.
PCT/GB2018/050613 2017-03-10 2018-03-09 Procédé de restauration de l'efficacité d'un agent antibactérien Ceased WO2018162928A1 (fr)

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JP2023508979A (ja) * 2019-12-23 2023-03-06 アルマ マータ ストゥディオールム－ウニヴェルシタ ディ ボローニャ 溶血性尿毒症症候群を処置するための化合物
JP7677977B2 (ja) 2019-12-23 2025-05-15 アルマ マータ ストゥディオールム－ウニヴェルシタ ディ ボローニャ 溶血性尿毒症症候群を処置するための化合物

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