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WO2006045156A1 - Traitement de micro-organismes a resistance pleiotrope - Google Patents

Traitement de micro-organismes a resistance pleiotrope Download PDF

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Publication number
WO2006045156A1
WO2006045156A1 PCT/AU2005/001668 AU2005001668W WO2006045156A1 WO 2006045156 A1 WO2006045156 A1 WO 2006045156A1 AU 2005001668 W AU2005001668 W AU 2005001668W WO 2006045156 A1 WO2006045156 A1 WO 2006045156A1
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Prior art keywords
antimicrobial peptide
colistin
macrolide
microorganism
drug resistant
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PCT/AU2005/001668
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English (en)
Inventor
Jian Li
Craig Robert RAYNOR
Roger Leigh NATION
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Monash University
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Monash University
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Priority claimed from AU2004906272A external-priority patent/AU2004906272A0/en
Application filed by Monash University filed Critical Monash University
Publication of WO2006045156A1 publication Critical patent/WO2006045156A1/fr
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    • 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
    • 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
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents

Definitions

  • This invention relates to pharmaceutical preparations with antimicrobial and anti-pathogenic activity against multi-drug resistant microorganisms, in particular, multi ⁇ drug resistant gram-negative microorganisms.
  • the invention particularly targets polymyxin-resistant bacteria such as Pseudomonas aeruginosa..
  • polymyxin-resistant P. aeruginosa has been isolated from patients 10 with eye infections, 9 ear infections, 7 and particularly in the sputum of patients with cystic fibrosis 11 ' 12 ' 13 .
  • the appearance of polymyxin-resistant gram-negative pathogens is of great concern.
  • the present invention addresses the need for an effective therapy against multi-drug resistant microorganisms. It also specifically addresses multi-drug resistant gram-negative bacteria. Examples of these are polymyxin-resistant gram-negative bacteria such as P. aeruginosa, and A. baumannii.
  • the present invention also addresses the problem of pathogenic factors released by living microorganisms in a host which lead to destructive effects on tissues at the site(s) of infection.
  • the present invention provides a pharmaceutical or veterinary composition
  • a pharmaceutical or veterinary composition comprising an antimicrobial peptide and a macrolide component together with a pharmaceutically or veterinarily acceptable carrier, wherein said antimicrobial peptide and macrolide component interact synergistically against a multi-drug resistant microorganism.
  • the present invention also provides a pharmaceutical or veterinary composition, comprising an antimicrobial peptide and a macrolide component together with, a pharmaceutically or veterinarily acceptable carrier, wherein said antimicrobial peptide and macrolide component interact with bi-directional antimicrobial synergy against a multi-drug resistant microorganism.
  • the pharmaceutical or veterinary composition may conveniently foe in the form of a kit in which the antimicrobial peptide and macrolide component axe held separately for separates, sequential or simultaneous use, It has been found that the antimicrobial peptide and macrolide component are effective in preventing or inhibiting growth of a multi-drug resistant microorganism.
  • the invention further provides the use of an antimicrobial peptide and a macrolide component, for preventing or inhibiting growth of a multi-drug resistant microorganism.
  • the invention still further provides a method of preventing or inhibiting growth of a multi-drug resistant microorganism comprising the step of administering to a subject in need thereof an effective amount of an antimicrobial peptide and a macrolide component.
  • the method above includes the administration of an effective amount of an antimicrobial peptide and a macrolide component separately, simultaneously or sequentially to a subject in need thereof.
  • It also provides a method of killing and/or preventing or inhibiting growth of a multi-drug resistant microorganism comprising the step of administering to a subject in need thereof an effective amount of a pharmaceutical or veterinary composition comprising an antimicrobial peptide and a macrolide component together with a pharmaceutically or veterinarily acceptable carrier, wherein said antimicrobial peptide and macrolide component interact synergisti ⁇ ally against a multi-drug resistant microorganism.
  • the invention also provides the use of an antimicrobial peptide and a macrolide component, in the manufacture of a medicament, for preventing or inhibiting growth of a multi-drug resistant microorganism.
  • an antimicrobial peptide in the manufacture of a medicament for preventing or inhibiting growth of a multi-drug resistant microorganism in a subject being treated with a medicament comprising a macrolide component.
  • a macrolide component in the manufacture of a medicament for preventing or inhibiting growth of a multi-drug resistant microorganism in a subject being treated with a medicament comprising an antimicrobial peptide.
  • an antimicrobial peptide and a macrolide component in the manufacture of a medicament for the killing, prevention or inhibition of growth of a multi-drug resistant microorganism, wherein said antimicrobial peptide and macrolide component interact synergistically against said multi-drug resistant microorganism.
  • being treated includes a subject who has been treated, is currently being treated and/or is going to be treated.
  • antimicrobial peptide and macrolide component are effective in the treatment and/or prophylaxis of an infection caused by a multi-drug resistant microorganism.
  • the present invention also provides the use of an antimicrobial peptide and macrolide component in the treatment and/or prophylaxis of an infection caused by a multi-drug resistant microorganism.
  • the invention still further provides a method for treatment and/or prophylaxis of an infection caused by a multi-drug resistant microorganism comprising the step of administering to a subject in need thereof an effective amount of an antimicrobial peptide and a macrolide component.
  • the method above includes the administration of an effective amount of an antimicrobial peptide and macrolide component separately, simultaneously or sequentially to a subject in need thereof.
  • It also provides a method of treatment and/or prophylaxis of an infection caused by a multi-drug resistant microorganism comprising the step of administering to a subject in need thereof an effective amount of a pharmaceutical or veterinary composition comprising an antimicrobial peptide and a macrolide component together with a pharmaceutically or veterinarily acceptable carrier, wherein ⁇ aid antimicrobial peptide and macrolide component interact synergistically against a multi-drug resistant microorganism.
  • the invention also provides the use of an antimicrobial peptide and a macrolide component in the manufacture of a medicament for the treatment and/or prophylaxis of an infection caused by a multi-drug resistant microorganism.
  • an antimicrobial peptide and a macrolide component in the manufacture of a medicament for the treatment or prophylaxis of an infection caused by a multi-drug resistant microorganism, wherein said antimicrobial peptide and macrolide component interact synergistically against said multi-drug resistant microorganism.
  • an antimicrobial peptide in the manufacture of a medicament for the treatment and/or prophylaxis of an infection caused by a multi-drug resistant microorganism in a subject being treated with a medicament comprising a macrolide component.
  • a macrolide component in the manufacture of a medicament for the treatment and/or prophylaxis of an infection caused by a multi-drug resistant microorganism in a subject being treated with a medicament comprising an antimicrobial peptide.
  • infections referred to above include infections selected from the group consisting of bacterial wound infections, mucosal infections, enteric infections, septic conditions, infections in airways, cerebrospinal fluid, blood, eyes, ears and skin.
  • the multi-drug resistant organisms are multi-drug resistant gram-negative microorganisms.
  • the multi-drug resistant microorganism is resistant to at least one member of the polymyxin class of antibiotics and synthetic derivatives thereof. polymyxin class of antibiotics and synthetic derivatives thereof.
  • the multi-drug resistant microorganism is polymyxin resistant Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, Salmonella spp, Klebsiella pneumonia, and/or Shigella spp.
  • antimicrobial peptide and macrolide component are effective in increasing the established anti-pathogenic effects of macrolides alone.
  • the present invention provides a pharmaceutical or veterinary composition
  • a pharmaceutical or veterinary composition comprising an antimicrobial peptide and a macrolide component, together with a pharmaceutically or veterinarily acceptable carrier, wherein said antimicrobial peptide enhances the anti-pathogenic activity of the macrolide component.
  • the invention further provides the use of an antimicrobial peptide and a macrolide component for inhibiting or preventing production of at least one pathogenic factor by a microorganism.
  • the invention further provides a method of inhibiting or preventing production of at least one pathogenic factor by a microorganism comprising the step of administering to a subject in need thereof an effective amount of an antimicrobial peptide and a macrolide component.
  • the invention further provides the use of an antimicrobial peptide and a macrolide component in the manufacture of a medicament for inhibiting or preventing production of at least one pathogenic factor by a microorganism.
  • an antimicrobial peptide in the manufacture of a medicament for inhibiting or preventing production of a pathogenic factor by a microorganism in a subject being treated with a medicament comprising a macrolide component. It also provides the use of a macrolide component in the manufacture of a medicament for inhibiting or preventing production of a pathogenic factor by a microorganism in a subject being treated with a medicament comprising an antimicrobial peptide.
  • the antimicrobial peptide has effective antipathogenic effects against a polymyxin- susceptible and -resistant microorganism.
  • the present invention provides a pharmaceutical or veterinary composition
  • a pharmaceutical or veterinary composition comprising an antimicrobial peptide, together with a pharmaceutically or veterinarily acceptable carrier, wherein, said antimicrobial peptide inhibits or prevents production of a pathogenic factor by a microorganism.
  • the invention further provides the use of an antimicrobial peptide for inhibiting or preventing production of a. pathogenic factor by a microorganism.
  • the invention further provides a method of inhibiting or preventing production of a pathogenic factor by a microorganism comprising the step of administering to a subject in need thereof an effective amount of an antimicrobial peptide.
  • an antimicrobial peptide in the manufacture of a medicament for inhibiting or preventing production of a pathogenic factor by a microorganism.
  • the microorganism referred to in this aspect of the invention is resistant to said antimicrobial peptide.
  • the pathogenic factors referred to above include production of biofilm, alginate production, expression of flagellin, cytokine production, alteration of polymorphonuclear cell function , and/or pyocyanin production.
  • the terms "antimicrobial peptide” and “macrolide component” as used herein include pharmaceutically acceptable salts or derivatives, pro-drugs, tautomers and/or isomers thereof.
  • an antimicrobial peptide includes mixtures of antimicrobial peptides
  • a macrolide component includes mixtures of two or more such components, and the like.
  • microorganism includes any microscopic organism or taxonomically related macroscopic organism within the categories algae, bacteria, fungi, yeast and . protozoa or the like. It includes susceptible and resistant microorganisms. Examples of infections produced by such microorganisms are provided herein.
  • microorganisms targeted in the first aspect of the present invention the prevention or inhibition of growth of, and/or the treatment and/or prophylaxis of an infection caused by, multi-drug resistant microorganisms.
  • gram-negative microorganisms are targeted.
  • the anti-pathologenic aspects of the invention target the broader class of "microorganism" as defined herein.
  • the antimicrobial peptide and macrolide component in the context of the anti-pathogenic aspect of the invention is most suited to treating a multi-drug resistant microorganism.
  • microorganism alone is not limited to “multi-drug resistant organism”, and encompasses both drug-susceptible and drug-resistant microorganisms.
  • multi-drug resistant microorganism refers to those organisms that are, at the very least, resistant to more than two antibiotics in different antibiotic classes. This includes those microorganisms that have more resistance i.e. those that are resistant to three or more antibiotics in a single antibiotic class. This also includes microorganisms that are resistant to a wider range of antibiotics, i.e. microorganisms that are resistant to one or more classes of antibiotics.
  • MIC breakpoints for an antibiotic are determined by the MIC distributions of pathogens in a clinical indication and its pharmacokinetics and pharmacodynamics in humans. While a microorganism may literally be susceptible to a high concentration of an antibiotic in vitro, the microorganism may in fact be resistant to that antibiotic at physiologically realistic concentrations.
  • the concentration of drug required to inhibit growth of or kill the microorganism is greater than the concentration that can safely be achieved without toxicity to the subject, the microorganism is considered to be resistant to the antibiotic
  • NCLS National Committee for Clinical Laboratory Standards
  • MIC values indicate resistance or susceptibility of a microorganism.
  • a suitable multi-drug resistant microorganism is an antimicrobial peptide resistant organism and/or a macrolide component resistant microorganism.
  • An antimicrobial peptide resistant microorganism is a microorganism resistant to at least the antimicrobial peptide being used against it in the invention.
  • a macrolide component resistant microorganism is a microorganism resistant to at least the macrolide component being used against it in the invention.
  • the multi-drug resistant microorganism is selected from the class of gram-negative microorganisms.
  • the antimicrobial peptide is a member of the polymyxin class of antibiotics.
  • polymyxin resistant refers to those microorganisms that are resistant to the member of the polymyxin class of antibiotics being used in the embodiment.
  • prevent or inhibit growth of a multi-drug resistant microorganism refers to the interference with growth or replication of the microorganism, which can include but does not necessarily extend to killing of the microorganism.
  • treatment and/or prophylaxis refers generally to affecting a subject, tissue or cell to obtain a desired pharmacologic and/or physiologic effect.
  • the effect may be prophylactic in terms of completely or partially preventing a disease or sign or symptom thereof, and/or may be therapeutic in terms of. a partial or complete cure of a disease.
  • the term “synergy” refers to the total increase in activity of both antimicrobial components over their additive antimicrobial activity. It includes the increase in activity of only one of the antimicrobial components.
  • the antimicrobial peptide and macrolide component show at least synergistic anti- pathogenic activity.
  • bidirectional synergy refers to the increase in activity of each antimicrobial component when used in conjunction with the other antimicrobial component, and not merely an increase in activity of one of the antimicrobial components.
  • the antimicrobial peptide and macrolide component show at least synergistic antimicrobial activity.
  • the antimicrobial peptide and macrolide component show bidirectional synergistic antimicrobial activity.
  • the antimicrobial peptide and macrolide component have surprisingly been found to act synergistically to reverse the substantial resistance to each of these when used alone against a multi-drug resistant microorganism, in particular, a multi-drug resistant gram-negative microorganism, including polymyxin resistant bacteria, such as polymyxin resistant P. aeruginosa.
  • a multi-drug resistant gram-negative microorganism including polymyxin resistant bacteria, such as polymyxin resistant P. aeruginosa.
  • an antimicrobial peptide such as colistin and a macrolide component such as erythromycin
  • antimicrobial peptides such as polymyxins have a permeabilising effect on the bacterial outer membrane of gram-negative microorganisms. Without wishing to be bound by theory, it is hypothesized that this permeabilising effect allows greater access to less water-soluble antibiotics such as macrolides which are not themselves active against certain gram-negative microorganisms.
  • synergistic activity is obtained in a multi-drug resistant microorganism, particularly a multi-drug resistant gram-negative microorganism such as a polymyxin resistant "microorganism," (some highly resistant isolates with MIC of colistin (sulfate) ⁇ 128 mg/L) e.g. P. aeruginosa.
  • a macrolide or macrolide component would be useful against a microorganism such as polymyxin-resistant bacteria, particularly given that the macrolide or macrolide component is not generally active against a microorganism such as polymyxin-susceptible gram-negative bacteria.
  • the minimum inhibitory concentration (MIC) mentioned. above is the lowest concentration of antibiotic at which, with the inoculum of 10 5 to 10 6 CFU/ml, there is no visible growth after 18 to 24h incubation at 35°C.
  • anti-pathogenic refers to activity in inhibiting or preventing production of a pathogenic factor released by living microorganisms in a host which leads to destructive effects of tissues at the site(s) of infection. This includes the inhibition of the production of biofilm by disruption of a quorum sensing system 14 , and consequent inhibition of alginate production, inhibition of the expression of flagellin in P. aeruginosa., perturbing of cytokine production and altering action of polymorphonucteer cell functions in vivo and ex vivo, and/or the inhibition of production of pyocyanin, which is produced by microorganisms such as P. aeruginosa and is a blue pigment which disrupts human ciliary beating in vitro 15 , inhibits epidermal cell growth 15 and also impedes lymphocyte proliferation.
  • biofilm Some bacteria, including P. aeruginosa, avidly form tightly arranged multi-cell structures in vivo known as biofilm.
  • the production of biofilm is important for the persistence of infectious processes such as seen in pseudomonal lung infections in patients with cystic fibrosis and diffuse panbronchiolitis and many other diseases.
  • Biofilm is resistant to phagocytosis by host immune cells and the effectiveness of antibiotics at killing bacteria in biofilm structures may be reduced by 10 to 1000 fold.
  • Biofilm production and arrangement is governed by quorum sensing systems.
  • the disruption of the quorum sensing system in bacteria such as P. aeruginosa is an important anti-pathogenic activity as it disrupts the biofilm formation and also inhibits alginate production.
  • Alginate induces antigen-antibody reactions and increases the fluid viscosity in the airways of CF patients.
  • the anti-pathogenic effect of alginate inhibition is beneficial by reducing the inflammation response and increasing phagocytosis as well as the ability for antibiotics to reach sensitive bacteria, resulting in enhanced clearance of bacteria.
  • compositions are therefore useful in the treatment of conditions associated with microbial infections.
  • antimicrobial peptide refers to peptides which have antimicrobial activity.
  • the polymyxin class is one class of antimicrobial peptides.
  • the term "polymyxin” is used in its broadest sense to encompass all members of the well known polymyxin class of antibiotics and synthetic derivatives thereof. Derivatives within this class are the non-cyclic derivatives of cyclic polymyxins, derivatives containing amino acid variations, derivatives containing substitutes of the fatty acid components with other fatty acids or substituents, derivatives with D- and L- amino acid conversions, and derivatives substituted with any one or more optional substituents identified below.
  • Classic polymyxins include polymyxin A, B1, B2, C, D1, D2, E1 and/or E2, F, G, M, P, S and T.
  • the polymyxins are cationic detergents and are relatively simple basic peptides with molecular masses of about 1000-1200 daltons.
  • members of the polymyxin class are polymyxin B (B 1 and B 2 ) and polymyxin E (colistin A and B) . Both these members include a cyclic heptapeptide ring with a tripeptide side chain. It is envisaged that declyclisation of the ring may result in a peptide with effective antimicrobial activity.
  • non-cyclic derivatives of the polymyxins and similar peptides are encompassed within the term "antimicrobial peptide” .
  • antiimicrobial peptide also included within the scope of "antimicrobial peptide” are all the components of polymyxin B and polymyxin E, as well as synthetic derivatives thereof.
  • polymyxin B (B 1 and B 2 ) is as follows:
  • Dab ⁇ , ⁇ -diaminobutyric acid, wherein ⁇ and ⁇ indicate the respective -NH 2 involved in the peptide linkage.
  • Polymyxin E (colistin) has many different components.
  • Dab ⁇ , ⁇ -diaminobutyric acid wherein ⁇ and ⁇ indicate the respective -NH 2 involved in the peptide linkage. It is envisaged that variation of these components, for example, by substituting a D-amino acid residue for the same or different L-amino acid residue or vice versa, varying the R substituents and/or conservative amino acid substitutions, while maintaining the synergistic antimicrobial activity with the macrolide component of the invention, is encompassed within the scope of the invention.
  • Minor components of colistin include the polymyxin E 3 and E 4 , norvaline-polymyxin E 1 , valine-polymyxin E 1 , and valine-polymyxin E 2 , isoleucine-polymyxin E 1 , isoleucine- polymyxin E 1 , polymyxin E 7 and isoleucine-polymyxin E 0 .
  • the antimicrobial peptide comprises any one or more components of colistin, further preferably colistin A and/or colistin B.
  • the proportion of colistin A and colistin B in commercial material varies between pharmaceutical suppliers and batches, but it is generally between 4.5:1 to 0.9:1.
  • Colistin is available commercially in two forms, colistin sulphate and sodium colistin methanesulphonate.
  • Sodium colistin methanesulphonate hydrolyses in aqueous media and forms a complex mixture of partially sulphomethylated derivatives plus colistin.
  • One or more of the above forms of colistin or its derivatives are encompassed within the scope of the invention.
  • antimicrobial peptide comprises salts of colistin, preferably salts of pharmaceutically acceptable cations such as sodium, potassium, lithium and the like, acid addition salts of pharmaceutically acceptable inorganic acids such as hydrochloric, orthophosphoric, sulphuric and the like, and/or salts of pharmaceutically acceptable organic acids such as acetic, propionic, methanesulphonic, and the like.
  • antimicrobial peptide comprises colistin methanesulphonate and/or colistin sulphate. In a further preferred embodiment, antimicrobial peptide comprises colistin sulphate.
  • amino acid within the scope of the present invention is used in its broadest sense and is meant to include naturally occurring L ⁇ -amino acids or residues.
  • the commonly used one and three letter abbreviations for naturally occurring amino acids are used herein. 17
  • the term includes D-amino acids as well as chemically modified amino acids such as amino acid analogs, naturally occurring amino acids that are not usually incorporated into proteins such as norleucine, and chemically synthesized compounds having properties known in the art to be characteristic of an amino acid.
  • analogs or mimetics of phenylalanine or proline, which allow the same conformational restriction of the peptide compounds as natural Phe or Pro are included, within the definition of amino acid.
  • Such analogs and mimetics are referred to herein as "functional equivalents" of an amino acid.
  • Other examples of amino acids are listed by Roberts and Vellaccio. 18 which is incorporated herein by reference.
  • Peptides synthesized by, for example, standard solid phase synthesis techniques are not limited to amino acids encoded by genes. Commonly encountered amino acids which are not encoded by the genetic codes, include, for example, those described in International Publication No.
  • WO 90/01940 such as, for example, 2-amino adipic acid (Aad) for GIu and Asp; 2-aminopimelic acid (Apm) for GIu and Asp; 2-aminobutyric (Abu) acid for Met, Leu, and other aliphatic amino acids; 2-aminoheptanoic acid (Ahe) for Met, Leu and other aliphatic amino acids; 2- aminoisobutyric acid (Aib) for GIy; cyclohexylalanine (Cha) for VaI, and Leu and Ile; homoarginine (Har) for Arg and Lys; 2,3-diaminopropionic acid (Dpr) for Lys, Arg and His; N-ethylglycine (EtGIy) for GIy, Pro, and Ala; N- ethylglycine (EtGIy) for GIy, Pro, and Ala; N- ethylasparigine
  • Conservative amino acid substitutions are shown in Table 1 under the heading of "exemplary substitutions” and “preferred substitutions” . If preferred substitutions do not result in a decrease or change in antimicrobial activity, then more substantial changes, denominated "exemplary substitutions" in Table 1, or as further described herein, may be introduced and the products tested for antimicrobial activity.
  • the term "macrolide component” refers to a component having a lactone ring and which has antimicrobial activity.
  • the term “macrolide” includes the well known class of macrolide antibiotics, including naturally occurring and synthetic derivatives thereof.
  • Classic macrolides are the 14-15 lactone ring-based compounds to the 16-19 membered lactone ring-based compounds to which is attached one or more deoxy sugars, and which has antimicrobial activity. Examples of these are erythromycin, clarithromycin and azithromycin, included in the term “macrolide” are lincosamides (clindamycin), azalides (azithromycin) and ketolides (telithromycin) .
  • erythromycin and clarithromycin differ from each other in that erythromycin and clarithromycin contain a 14-membered lactone ring and azithromycin contains a 15-membered lactone ring.
  • Clarithromycin differs from erythromycin only by methylation of the hydroxyl group at the 6-position, and azithromycin differs by the addition of a methyl substituted nitrogen atom into the lactone ring.
  • the macrolide lactone ring may be optionally substituted with various organic substituents.
  • optionally substituted means that one or more members of the lactone ring or a substituent on the lactone ring may or may not be further substituted with, one or more groups selected from alkyl, alkenyl, alkynyl, aryl, halo, haloalkyl, haloalkenyl, haloalkynyl, haloaryl, hydroxy, alkoxy, alkenyloxy, .
  • Clarithromycin and colistin sulphate are preferred over erythromycin and colistin sulphate.
  • Clarithromycin and colistin methanesulphonate are preferred over erythromycin and colistin methanesulphonate.
  • Colistin sulphate and a macrolide component are preferred over colistin methanesulphonate and a macrolide component as they showed 80% lower FlC values, thus expressing greater synergy. Therefore colistin sulphate and clarithromycin are preferred over colistin methanesulphonate and erythromycin.
  • the antimicrobial peptide and/or the macrolide may be used in the form of their salts, derivatives, pro-drugs, tautomers and/or isomers thereof.
  • the salts of the antimicrobial peptide and macrolide component are preferably pharmaceutically acceptable, but it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the present invention since they are useful as intermediates in the preparation of pharmaceutically acceptable salts.
  • Examples of pharmaceutically acceptable salts include salts of pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, ammonium and alkylammonium; acid addition salts of pharmaceutically acceptable inorganic acids such as hydrochloric, orthophosphoric, sulphuric, phosphoric, nitric, carbonic, boric, sulfamic and hydrobromic acids; or salts of pharmaceutically acceptable organic acids except as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, trihalomethanesulphonic, toluenesulphonic, benzenesulphonic, salicylic, sulphanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, as
  • antimicrobial peptides and/or macrolide components of the present invention may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention.
  • pharmaceutically acceptable derivative is meant any pharmaceutically acceptable salt, hydrate or any other compound which, upon administration to the subject, is capable of providing (directly or indirectly) an antimicrobial peptide and/or macrolide component or residue thereof.
  • pro-drug is used herein in its broadest sense to include those compounds which are converted in vivo to antimicrobial peptides and/or macrolide components of the present invention.
  • tautomer is used herein in its broadest sense to include antimicrobial peptides and/or macrolide components which are capable of existing in a state of equilibrium between two isomeric forms. Such compounds may differ in the bond connecting two atoms or groups and the position of these atoms or groups in the compound.
  • antimicrobial peptides and/or macrolide components may have one or more chiral centres, they are capable of existing in enantiomeric forms.
  • microorganism includes any microscopic organism or taxonomically related macroscopic organism within the categories algae, bacteria, fungi, yeast and protozoa, or the like.
  • the microorganisms targeted in the first aspect of the present invention are multi-drug resistant microorganisms. Preferably, gram-negative microorganisms are targeted.
  • Bacterial infections include, but are not limited to, infections caused by Bacillus cereus, Bacillus anthracis, Clostridium botulinum, Clostridium difficile, Clostridium tetani, Clostridium perfringens, Corynebacteria diphtheriae, Enterococcus (Streptococcus D) , Listeria monocytogenes, Pneumococcal infections (Streptococcus pneumoniae) , Staphylococcal infections and Streptococcal infections ; Gram-negative bacteria including Bacteroides, Bordetella pertussis, Brucella, Campylobacter infections, enterohaemorrhagic Escherichia coli (EHEC/E.
  • EHEC/E enterohaemorrhagic Escherichia coli
  • coli 0157 H7) enteroinvasive Escherichia, coli (EIEC) , enterotoxigenic Escherichia coli (ETEC) , Haemophilus influenzae, Helicobacter pylori, Klebsiella, pneumoniae, Legionella, spp., Moraxella catarrhalis, Neisseria gonnorrhoeae, Neisseria meningitidis, Proteus spp., Pseudomonas aeruginosa, Salmonella spp., Shigella spp.,
  • Vibrio cholera and Yersinia acid fast bacteria including Mycobacterium tuberculosis, Mycobacterium avium- intracellulars, Myobacterium johnei, Mycobacterium leprae, atypical bacteria, Chlamydia, Mycoplasma, Rickettsia, Spirochetes, Treponema pallidum, Borrelia recurrentis,
  • the microbial infection is caused by gram-negative bacterium, for example, P. aeruginosa., A. baumannii, Salmonella spp, Klebsiella pneumonia, Shigella spp. and/or Stenotrophomonas maltophilia.
  • gram-negative bacterium for example, P. aeruginosa., A. baumannii, Salmonella spp, Klebsiella pneumonia, Shigella spp. and/or Stenotrophomonas maltophilia.
  • microbial infections include bacterial wound infections, mucosal infections, enteric infections, septic conditions, pneumonia, trachoma, ornithosis, trichomoniasis and salmonellosis, especially in veterinary practice.
  • infections caused by P. aeruginosa include:
  • infections caused by A. baumannii include: Nosocomial infections; 1. Bacteraemia and sepsis; 2. Respiratory tract infections in mechanically ventilated patients; 3. Post-surgery infections on invasive devices;
  • infections caused by Stenotrophomonas maltophilia include bacteremia, pneumonia, meningitis, wound infections and urinary tract infections.
  • Some hospital breaks are caused by contaminated disinfectant solutions, respiratory devices, monitoring instruments and ice machines. Infections usually occur in debilitated patients with impaired host defense mechanisms.
  • infections caused by Klebsiella pneumoniae include community-acquired primary lobar pneumonia, particularly in people with compromised pulmonary function and alcoholics. It also caused wound infections, soft tissue infections and urinary tract infections.
  • infections caused by Salmonella, spp. are acquired by eating contaminated food products. Infections include enteric fever, enteritis and. bacteremia. Examples of infections caused, by Shigella, spp. include gastroenteritis (shigellosis) .
  • antimicrobial peptides and/or macrolide components of the invention may also be used in various fields where antiseptic treatment or disinfection of materials is required, for example, surface disinfection.
  • subject refers to any animal having a disease or condition which requires treatment with a pharmaceutically-active agent.
  • the subject may be a mammal, preferably a human, or may be a domestic or companion animal. While it is particularly contemplated that the antimicrobial peptides and/or macrolide components of the invention are suitable for use in medical treatment of humans, it ie also applicable to veterinary treatment, including treatment of companion animals such as dogs and cats, and domestic animals such as horses, ponies, donkeys, mules, llama, alpaca, pigs, cattle and sheep, or zoo animals such as primates, felids, canids, bovids, and ungulates.
  • Suitable mammals include members of the orders Primates, Rodentia, Lagomorpha, cetacea, Carnivora, Perissodactyla and Artiodactyla. Members of the orders Perissodactyla and Artiodactyla are particularly preferred because of their similar biology and economic importance.
  • Artiodactyla comprises approximately 150 living species distributed through nine families: pigs (Suidae) , peccaries (Tayassuidae) , hippopotamuses (Hippopotamidae) , camels (Camelidae) , chevrotains (Tragulidae) , giraffes and okapi (Giraffidae) , deer (cervidae) , pronghorn (Antilocapridae) , and cattle, sheep, goats and antelope (Bovidae) . Many of these animals are used as feed animals in various countries.
  • the term "effective amount” is meant an amount of antimicrobial peptides and/or macrolide components of the present invention effective to yield a desired antibiotic activity.
  • the specific "effective amount” will, obviously, vary with such factors as the particular condition being treated, the physical condition of the subject, the type of subject being treated, the duration of the treatment, the nature of concurrent therapy (if any) , and the specific formulations employed, the ratio of the antimicrobial peptides and/or macrolide components to each other, the structure of each of these components or their derivatives.
  • the antimicrobial peptides and/or macrolide components of the present invention may additionally be combined with other medicaments to provide an operative combination. It is intended to include any chemically compatible combination of pharmacologically-active agents, as long as the combination does not eliminate the activity of the antimicrobial peptides and/or macrolide components. It will be appreciated that the antimicrobial peptides and/or macrolide components of the invention and the other medicament may be administered separately, sequentially or simultaneously.
  • medicaments which may be used when treating bacterial infections include salbutamol, ipratropium, dornase alpha, for example, for use in inhalation for respiratory infections such as cystic fibrosis.
  • a “pharmaceutical carrier” is a pharmaceutically acceptable solvent, suspending agent or vehicle for delivering the combination of antimicrobial peptides and/or macrolide components to the subject.
  • the carrier may be liquid or solid and is selected with the planned manner of administration in mind.
  • Each carrier must be pharmaceutically “acceptable” in the sense of being compatible with other ingredients of the composition and non injurious to the subject.
  • the antimicrobial peptides and/or macrolide components may be administered topically, including local delivery to the gastrointestinal tract and other membrane surfaces including aerosol delivery for administration to lungs or nasal cavity, parenterally or orally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, and vehicles .
  • parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrathecal, intraventricula, intracranial, injection or infusion techniques.
  • Suitable disintegrating agents include corn starch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar.
  • Suitable flavouring agents include peppermint oil, oil of wintergreen, cherry, orange or raspberry flavouring.
  • Suitable preservatives include sodium benzoate, vitamin E, alphatocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite.
  • Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc.
  • Suitable time delay agents include glyceryl monostearate or glyceryl distearate.
  • the tablets contain the active combination in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets.
  • excipients may be, for example, (1) inert diluents, such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating and disintegrating agents, such as corn starch or alginic acid; (3) binding agents, such as starch, gelatin or acacia; and (4) lubricating agents, such as magnesium stearate, stearic acid or talc.
  • inert diluents such as calcium carbonate, lactose, calcium phosphate or sodium phosphate
  • granulating and disintegrating agents such as corn starch or alginic acid
  • binding agents such as starch, gelatin or acacia
  • lubricating agents such as magnesium stearate, stearic acid or talc.
  • These tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.
  • a time delay material such as gly
  • Coating may also be performed using techniques described in the U.S. Pat. Nos. 4,256,108; 4,160,452; and 4,265,874 to form osmotic therapeutic tablets for control release.
  • the antimicrobial peptides and/or macrolide components can be administered, for in vivo application, parenterally by injection or by gradual infusion over time independently or together. Administration may be intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, intracavity, transdermal, inhalation, intracisternal, intraventricular, interathecal (intra-CSF) or infusion by, for example, infusion pump. Also included are ear drops, eye drops, gels for skin infections etc.
  • Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions.
  • non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.
  • Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media.
  • Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride
  • lactated Ringer's intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose) , and the like.
  • Preservatives and other additives may also be present such as, for example, anti-oxidants, chelating agents, growth factors and inert gases and the like.
  • Treating covers any treatment of, or prevention of disease in a vertebrate, a mammal, particularly a human, and includes: (a) preventing the disease from occurring in a subject that may be predisposed to the disease, but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; or (c) relieving or ameliorating the effects of the disease, i.e., cause regression of the effects of the disease.
  • the invention includes various pharmaceutical compositions useful for ameliorating disease.
  • the pharmaceutical compositions according to one embodiment of the invention are prepared by bringing antimicrobial peptides and/or macrolide components, analogues, derivatives or salts thereof, or combinations of these compounds and one or more other medicaments into a form suitable for administration to a subject using carriers, excipients and additives or auxiliaries.
  • carriers or auxiliaries include magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, milk protein, gelatin, starch, vitamins, cellulose and its derivatives, animal and vegetable oils, polyethylene glycols and solvents, such as sterile water, alcohols, glycerol and polyhydric alcohols .
  • Intravenous vehicles include fluid and nutrient replenishers.
  • Preservatives include antimicrobial, anti-oxidants, chelating agents and inert gases.
  • Other pharmaceutically acceptable carriers include aqueous solutions, non-toxic excipients, including salts, preservatives, buffers and the like, as described, for instance, in Remington's Pharmaceutical Sciences, 20th ed. Williams &. Williams (2000) , the British National Formulary, 43 rd edition (British Medical Association and Royal Pharmaceutical Society of Great Britain, 2000) , the contents of which are hereby incorporated by reference. The pH and exact concentration of the various components of the pharmaceutical composition are adjusted according to routine skills in the art. 19
  • the pharmaceutical compositions are preferably prepared and administered in dose units.
  • Solid dose units may be tablets, capsules and suppositories.
  • different daily doses can be used for treatment of a subject. Under certain circumstances, however, higher or lower daily doses may be appropriate.
  • the administration of the daily dose can be carried out both by single administration in the form of an individual dose unit or else several smaller dose units and also by multiple administration of subdivided doses at specific intervals.
  • dosages used in vitro may provide useful guidance in the amounts useful for in situ administration of the pharmaceutical composition, and animal models may be used to determine effective dosages for treatment of the bacterial infections.
  • Formulations for oral use may be in the form of hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.
  • Aqueous suspensions normally contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspension.
  • excipients may be (1) suspending agent such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; (2) dispersing or wetting agents which may be (a) naturally occurring phosphatide such as lecithin; (b) a condensation product of an alkylene oxide with a fatty acid, for example, polyoxyethylene stearate; (c) a condensation product of ethylene oxide with a long chain aliphatic alcohol, for example, heptadecaethylenoxycetanol; (d) a condensation product of ethylene oxide with a partial ester derived from a fatty acid and hexitol such as polyoxyethylene sorbitol monooleate, or (e) a condensation product of ethylene oxide with a
  • compositions may be in the form of a sterile injectable aqueous or oleagenous suspension.
  • This suspension may be formulated according to known methods using those suitable dispersing or wetting agents and suspending agents which have been mentioned above.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1, 3-butanediol.
  • a non-toxic parenterally-acceptable diluent or solvent for example, as a solution in 1, 3-butanediol.
  • acceptable vehicles and solvents that may be employed are water,
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono-or diglycerides.
  • fatty acids such as oleic acid find use in the preparation of injectables.
  • the antimicrobial peptides and/or macrolide components of the present invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles.
  • liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.
  • antimicrobial peptides and/or macrolide components of the present invention may also be presented for use in the form of veterinary compositions, which may be prepared, for example, by methods that are conventional in the art.
  • veterinary compositions include those adapted for:
  • oral administration external application, for example drenches (e.g. aqueous or non-aqueous solutions or suspensions) ; tablets or boluses; powders, granules or pellets for admixture with feed stuffs; pastes for application to the tongue;
  • drenches e.g. aqueous or non-aqueous solutions or suspensions
  • tablets or boluses e.g. aqueous or non-aqueous solutions or suspensions
  • pastes for application to the tongue for example drenches (e.g. aqueous or non-aqueous solutions or suspensions) ; tablets or boluses; powders, granules or pellets for admixture with feed stuffs; pastes for application to the tongue;
  • parenteral administration for example by subcutaneous, intramuscular or intravenous injection, e.g. as a sterile solution or suspension; or (when appropriate) by intramammary injection where a suspension or solution is introduced in the udder via the teat;
  • topical applications e.g. as a cream, gel, ointment or spray applied to the skin, delivery for local activity in the gut and lung, vagina, rectum, intrarectally as a suppository, cream or foam.
  • Dosage levels of the polymyxin and macrolide of the present invention may vary.
  • the amount of the active ingredients that may be combined with the carrier materials to produce a single dosage will vary depending upon the host treated and the particular mode of administration.
  • Figure 1 is a box and whisker plot for colistin sulphate in combination with macrolides against colistin- resistant and colistin-susceptible P. aeruginosa isolates demonstrating that total FICs were dramatically decreased (clarithromycin>erythromycin and resistant>sensitive) .
  • Figure 2 FIC colistin sulphate and FIC erythromycin in combination against colistin-resistant and colistin- susceptible P. aeruginosa isolates, demonstrating the substantial bidirectional synergy, and the greater synergy with colistin-resistant isolates.
  • Figure 3 FIC colistin sulphate &nd FIC clarithromycin in combination against colistin-resistant and colistin- susceptible P. aeruginosa isolates, demonstrating the substantial bidirectional synergy (greater synergy with clarithromycin compared to erythromycin in Figure 2) , and the greater synergy with colistin-resistant isolates.
  • Figure 4 Box and whisker plots for colistin methanesulphonate in combination with macrolides against colistin-resistant and colistin-susceptible P. aeruginosa isolates demonstrating that total FICs were dramatically decreased (clarithromycin>erythromycin and resistant>sensitive) .
  • Figure 5 FIC colistin methanesulphonate and FIC erythromycin in combination against colistin-resistant and colistin- susceptible P. aeruginosa isolates, demonstrating the substantial bidirectional synergy, and the greater synergy with colistin-resistant isolates.
  • Figure 6 FIC colistin methanesulphonate and FIC clarithromycin in combination against colistin-resistant and colistin- susceptible P. aeruginosa isolates, demonstrating the substantial bidirectional synergy, and the greater synergy with colistin-resistant isolates.
  • Figure 7 Inhibition of pyocyanin by the combination for P. aeruginosa ATCC27853 demonstrating substantial enhancement of erythromycin-induced anti-pathogenic effect at a colistin sulphate concentration of 0.5mg/L (or 1/2XMIC) .
  • Figure 8 Inhibition of pyocyanin by the combination for P. aeruginosa 19453 muc, demonstrating substantial enhancement of erythromycin-induced anti-pathogenic effects at increasing colistin sulphate concentrations at low fractions of the colistin sulphate MIC (MIC ⁇ 128 mg/L) .
  • Figure 9 Inhibition of pyocyanin by the combination for P. aeruginosa. 19626 muc demonstrating substantial enhancement of erythromycin-induced anti-pathogenic effects at increasing colistin sulphate concentrations at low fractions of the colistin sulphate MIC (MIC ⁇ 128 mg/L) , and substantial concentration-dependent anti- pathogenic effects of colistin sulphate alone (where erythromycin concentration was 0 mg/L.
  • Figure 10 FIC colistin sulphate and FIC erythromycin in combination against colistin-resistant and colistin- susceptible A. baumannii isolates, demonstrating the substantial bidirectional synergy, and the greater synergy with colistin-resistant isolates.
  • Figure 11 FIC colistin sulphate and FIC erythromycin in combination against colistin-resistant and colistin- susceptible Stenotrophomonas maltophilia isolates, demonstrating the bidirectional synergy.
  • Figure 12 FIC colistin methanesulphonate and FIC erythromycin in combination against colistin-resistant and colistin- susceptible A. baumannii isolates, demonstrating the modest bidirectional synergy, and the greater synergy with colistin-resistant isolates.
  • Figure 13 Killing kinetics of colistin sulphate/erythromycin against P. aeruginosa (Table 1) .
  • Figure 14 Killing kinetics of colistin sulphate /erythromycin and colistin methanesulphonate/erythromycin against P. aeruginosa (Tables 2 and 3) EXAMPLES
  • the stock solutions of colistin sulphate and colistin methanesulphonate were passed through a 0.22 ⁇ m filter.
  • the stock solution was diluted with cation-adjusted Mueller-Hinton broth (CAMHB) using the National Committee for Clinical Laboratory Standards (NCCLS) protocol.
  • FICs were determined at concentrations of 0.25 - 16 mg/L for colistin sulphate or 1.0 - 64 mg/L for colistin methanesulphonate with 0.125 - 128 mg/L for the macrolides using 96-well micro-plates.
  • the initial inocula of P. aeruginosa were 5x10 5 to 10 6 CFU/mL. Results were read at 24 h after incubation at 35°C.
  • FICs of combinations of colistin sulphate or colistin methanesulphonate with erythromycin were tested against 22 P. aeruginosa isolates including 12 colistin-resistant isolates.
  • FICs of combinations of colistin sulphate or colistin methanesulphonate with clarithromycin were tested against 10 P. aeruginosa isolates including 8 colistin-resistant isolates, a subset of those tested with erythromycin combinations described above.
  • FICs of combinations of colistin sulphate with erythromycin were tested against 5 Acinetobacter baumannii isolates including 3 colistin-resistant isolates.
  • FICs of combinations of colistin methanesulphonate with erythromycin were tested against 2 Acinetobacter baumannii isolates including 1 colistin-resistant isolates.
  • FICs of combinations of colistin sulphate with erythromycin were tested against 3 Stenotrophomonas maltophilia isolates including 2 colistin-resistant isolates .
  • the FIC was calculated by
  • MIC is the minimum inhibitory concentration defined as the lowest concentration without visible growth after overnight incubation at 35°C with an inoculum of 5x10 5 to 10 6 CFU/mL.
  • Killing curves were determined using a range of concentrations of colistin sulphate (or colistin methanesulphonate) together with a macrolide in a checkerboard design. Briefly, two days before an experiment, isolates stored at -80°C were cultured on horse blood agar and incubated at 35°C for 18 h for P. aeruginosa ATCC27853 (colistin susceptible and macrolide resistant) or 24 h for 20844 n/tnS (clinical isolate, resistant to both colistin and macrolide) . On the day before an experiment, 10 mL CAMHB was inoculated and incubated in a shaking water bath (80 cycles per minute) at 35°C.
  • colistin sulphate or colistin methanesulphonate
  • MST mean survival time (in hours)
  • AUMC area- under-the-curve of CFU/mL versus time multiplied by time of sampling in hours
  • AUC area-under-the-curve of CFU/mL versus time. Areas were determined using the trapezoidal rule.
  • results below demonstrate the effectiveness of the antimicrobial peptide/macrolide component combination as described.
  • In vivo testing can be conducted according to the following assay.
  • the assay is described with reference to the example of azithromycin, erythromycin and/or clarithromycin as the representative macrolide.
  • Colistin methane sulphonate and/or colistin will be used as the representative antimicrobial peptide.
  • Using the mouse thigh infection model 21 combinations of the representative macrolide and antimicrobial peptide will be examined against polymyxin-resistant P. aeruginosa.
  • the subcutaneous doses of colistin methanesulphonate and colistin will be designed to achieve clinically relevant plasma concentrations (approximately 0.4 - 25 mg/L for CMS and 0.2 - 3.5 mg/L for colistin) 22
  • the subcutaneous dose of macrolide will be based not only upon the results from in vitro studies but also on knowledge of clinically achievable plasma concentrations (for example, in the case of azithromycin, approximately 0.2 - 4.5 mg/L; Product Information, Pfizer) .
  • Both the antimicrobial peptide and the macrolide referred to throughout the specification are approved for administration to humans. Accordingly their toxicity profiles, bioavailability, half-life following administration and other relevant properties following administration in humans are well known to those in the art.
  • the appropriate optimal dosage of each of the antimicrobial peptide and macrolide component to treat a particular condition in a subject can be readily determined by those in the art on the basis of information available in the art and described herein.
  • a macrolide-group alone will not be included because it is intrinsically inactive against P. aeruginosa.
  • the mice will be killed and the thigh removed. Following homogenisation, counting for viable organisms will be conducted to conclude the antibacterial effect and the concentrations of the bacterial pathogenic factor, pyocyanin, will be determined by HPLC.
  • FIC clarithromycin and FIC erythromycin reached 0.0039 and 0.0078, respectively ( Figures 4-6) . According to standard definitions substantial bidirectional synergy was observed. Interestingly, clarithromycin and colistin methanesulphonate combinations showed even greater synergy than colistin methanesulphonate and erythromycin combinations.
  • Table 1 Killing kinetics of colistin sulphate/erythromycin against P. aeruginosa ATCC27853 (colistin-sensitive and macrolide-resistant)
  • Table 3 Killing kinetics of colistin methanesulphonate/erythromycin demonstrating significant yet weaker synergy than colistin sulphonate/erythromycin (Table 2) against P. aeruginosa 20844 n/mucS (colistin and macrolide-resistant

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Abstract

L'invention concerne une composition pharmaceutique ou vétérinaire renfermant un peptide anti-microbien et un composant macrolide, conjointement avec un excipient acceptable sur le plan pharmaceutique ou vétérinaire, le peptide anti-microbien et le composant macrolide interagissant de manière synergique contre un micro-organisme à résistance pléiotrope, une méthode de traitement et/ou de prophylaxie d'une infection engendrée par ledit micro-organisme, au moyen de la composition dans la fabrication d'un médicament destiné audit traitement.
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US20100028334A1 (en) * 2006-12-15 2010-02-04 Trustees Of Boston University Compositions and methods to potentiate colistin activity
WO2008076806A3 (fr) * 2006-12-15 2008-11-27 Univ Boston Compositions et procédés pour potentialiser l'activité de la colistine
WO2010103119A1 (fr) * 2009-03-13 2010-09-16 Da Volterra Compositions et procédés d'élimination de bactéries gram négatives
CN102413826A (zh) * 2009-03-13 2012-04-11 达·沃尔泰拉公司 用于消除革兰氏阴性菌的组合物和方法
CN103923190B (zh) * 2013-01-14 2018-04-03 上海医药工业研究院 从多黏菌素b的混合组分中分离纯化多黏菌素b1的方法
CN103923190A (zh) * 2013-01-14 2014-07-16 上海医药工业研究院 从多黏菌素b的混合组分中分离纯化多黏菌素b1的方法
USRE48335E1 (en) 2014-04-01 2020-12-01 Monash University Polymyxin derivatives as antimicrobial compounds
WO2015149131A1 (fr) * 2014-04-01 2015-10-08 Monash University Dérivés de polymyxine utilisés comme composés antimicrobiens
US10047126B2 (en) 2014-04-01 2018-08-14 Monash University Polymyxin derivatives as antimicrobial compounds
AU2015240435B2 (en) * 2014-04-01 2019-09-19 Monash University Polymyxin derivatives as antimicrobial compounds
WO2016100578A2 (fr) 2014-12-16 2016-06-23 Micurx Pharmaceuticals, Inc. Polymyxines antimicrobiennes pour le traitement d'infections bactériennes
US9771394B2 (en) 2014-12-16 2017-09-26 Micurx Pharmaceuticals, Inc. Antimicrobial polymyxins for treatment of bacterial infections
CN104888220A (zh) * 2015-06-03 2015-09-09 杨亮 一种有效降低铜绿假单胞菌生物膜耐药性的用药方法
US11225505B2 (en) 2015-09-29 2022-01-18 Monash University Antimicrobial polymyxin derivative compounds
WO2021150792A1 (fr) 2020-01-21 2021-07-29 Micurx Pharmaceuticals, Inc. Nouveaux composés et composition pour la thérapie ciblée de cancers associés au rein
CN112316112A (zh) * 2020-11-30 2021-02-05 河南农业大学 阿奇霉素在逆转食品动物源致病菌抗生素耐药性的用途
CN116509872A (zh) * 2023-03-29 2023-08-01 中国农业大学 抑制mst1/2或敲除巨噬细胞中的mst1/2在感染多黏菌素耐药细菌疾病治疗中的应用

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