WO2018102890A1 - Constructions de visualisation - Google Patents

Constructions de visualisation Download PDF

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Publication number
WO2018102890A1
WO2018102890A1 PCT/AU2017/051365 AU2017051365W WO2018102890A1 WO 2018102890 A1 WO2018102890 A1 WO 2018102890A1 AU 2017051365 W AU2017051365 W AU 2017051365W WO 2018102890 A1 WO2018102890 A1 WO 2018102890A1
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WIPO (PCT)
Prior art keywords
construct
component
visualization
microorganism
linker
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PCT/AU2017/051365
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English (en)
Inventor
Mark Blaskovich
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The University Of Queensland
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Publication date
Priority claimed from AU2016905101A external-priority patent/AU2016905101A0/en
Application filed by The University Of Queensland filed Critical The University Of Queensland
Publication of WO2018102890A1 publication Critical patent/WO2018102890A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • 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
    • A61K38/14—Peptides containing saccharide radicals; Derivatives thereof, e.g. bleomycin, phleomycin, muramylpeptides or vancomycin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0056—Peptides, proteins, polyamino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/088—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins conjugates with carriers being peptides, polyamino acids or proteins
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K17/00—Carrier-bound or immobilised peptides; Preparation thereof
    • C07K17/14—Peptides being immobilised on, or in, an inorganic carrier
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911—Bacteria
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • THE present invention relates to constructs comprising a glycopeptide antibiotic and a visualization component.
  • the invention relates to visualizing of microorganisms or components thereof bound to such constructs.
  • the invention also relates to use of such constructs for the diagnosis or monitoring of diseases related to bacterial infections.
  • Microorganisms such as bacteria cause a vast range of diseases or conditions of living organisms, including plants and animals such as humans. Since the middle of the last century, antibiotics have been effectively used for control of bacterial diseases. However, bacteria have substantial capacity to develop resistance to antibiotics, and disease caused by antibiotic resistant bacteria is currently considered a major global crisis.
  • Bacteraemia and sepsis are related conditions in which microorganisms (typically bacteria) are present in blood.
  • microorganisms typically bacteria
  • bacteraemia a microorganism enters the blood, e.g. through a wound, infection, or surgical procedure.
  • sepsis a microorganism enters and replicates in the blood.
  • Sepsis is typically considered a very serious condition, and is frequently life-threatening. Sepsis is commonly caused by the Gram positive cocci bacteria Staphylococcus species, particularly S. aureus.
  • Other Gram positive bacteria including Streptococcus, and Enterococcus species can cause sepsis, as can certain Gram negative bacteria (e.g. Escherichia, Pseudomonas, and Klebsiella species) and fungi, e.g. Candida species.
  • diagnosis of the site of infection leading to the sepsis can be important.
  • Vancomycin a glycopeptide antibiotic
  • Vancomycin is commonly used for treatment of infections by multidrug-resistant Gram-positive bacteria. Vancomycin is generally considered as the first-line treatment for serious MRS A infections.
  • resistance to glycopeptide antibiotics is being observed, for example in Enterococci species.
  • Reduced susceptibility of MRSA to glycopeptide antibiotics, known as glycopeptide-intermediate S. aureus (GISA) has also been observed and cases of vancomycin-resistant S. aureus (VRSA) have begun to be reported.
  • GISA glycopeptide-intermediate S. aureus
  • VRSA vancomycin-resistant S. aureus
  • Technologies used to visualize cellular structure and dynamics in living cells enable scientists to understand interaction and function of biomolecules within the complex system. In particular, understanding the cellular complexity of bacteria is important in developing new strategies to combat antibiotic -resistant bacteria. Additionally, such technologies may assist in identifying the site of infection in certain conditions such as sepsis.
  • the present invention is directed to constructs comprising an optionally derivatized glycopeptide antibiotic connected to a visualization component.
  • Constructs of this aspect may have the following general structure:
  • A is the optionally derivatized glycopeptide antibiotic
  • Li is the first linker
  • V is the visualization component.
  • the construct comprises:
  • the construct of this aspect is for binding to a microorganism, wherein the optionally derivatized glycopeptide antibiotic binds to the microorganism.
  • the microorganism is a Gram positive bacteria.
  • the visualization component facilitates visualization of a microorganism bound to the construct.
  • the visualization component is a light-emitting component.
  • the light emitting component emits light in the visual spectrum, i.e. in a wavelength range of about 400 nm to about 700 nm. In some embodiments, the light emitting component emits light in the infrared spectrum, i.e. in a wavelength range of about 700 nm to about 1mm.
  • the light emitting component is a luminescent component.
  • the light emitting component comprises a fluorescent compound.
  • the fluorescent compound is an organic or carbon-containing compound.
  • the organic fluorescent compound comprises one or more heterocyclic groups.
  • the heterocyclic groups are N or O-containing heterocyclic groups.
  • the fluorescent component is or comprises a quantum dot semiconductor particle.
  • the fluorescent compound has a neutral charge.
  • the fluorescent compound has a molecular weight between about 100 and about 500 Da. Preferably, the fluorescent compound has a molecular weight between about 150 and about 250 Da.
  • the fluorescent compound is optionally derivatized 7-nitrobenz-2-oxa-l,3-diazol-4-yl (NBD). In one particularly preferred embodiment, the fluorescent compound is optionally derivatized 7- (dimethylamino)-coumarin-4-acetic acid (DMACA). In one particularly preferred embodiment, the fluorescent compound is optionally derivatized 5-(dimethylamino)-l naphthalene- 1-sulfonyl (dansyl). In one particularly preferred embodiment, the fluorescent compound is optionally derivatized 4,4-difluoro-4-bora-3a,4a-diaza-s- indacene (BODIPY).
  • BODIPY 4,4-difluoro-4-bora-3a,4a-diaza-s- indacene
  • the fluorescent compound is optionally derivatized (E)-2-(2-(6-hydroxy-2,3-dihydro-lH-xanthen-4- yl)vinyl)-3,3-dimethyl- l-propyl-3H-indol-l-ium iodide ( ⁇ ).
  • the fluorescent compound is optionally derivatized (E)-3,3- dimethyl-2-(2-(6-(prop-2-yn- l-yloxy)-2,3-dihydro-lH-xanthen-4-yl)vinyl)- l-propyl- 3H-indol-l-ium iodide (PXPI).
  • the visualization component is a Magnetic Resonance Imaging (MRI) visualization component.
  • the MRI component comprises 19 F (e.g. as part of an organic compound, although without limitation thereto).
  • the MRI component comprises gadolinium, preferably Gd(III).
  • Gd(III) is of a macrocycle complex.
  • the visualization component is a Positron Emission Tomography (PET) visualization component.
  • PET Positron Emission Tomography
  • the PET visualization component comprises 64 Cu.
  • the PET visualization component comprises 68 Ga.
  • the 64 Cu or 68 Ga is contained within a macrocycle complex.
  • PET visualization component contains F. In some embodiments, the PET visualization component contains n C. In some embodiments, the PET visualization
  • the PET visualization component contains I.
  • the PET visualization component contains 99m Tc.
  • the 18 F; U C; 124 I; or 99m Tc as per these embodiments may be present as part of an organic compound, although without limitation thereto.
  • the PET visualization component may comprise a chelating compound.
  • the chelating compound is optionally derivatized l,4,7,10-tetraazacyclododecane- l,4,7-triyl)triacetic acid (DOT A), such as 2,2',2"-(10-(2-oxo-2-(prop-2-yn- l-ylamino)ethyl)- 1,4,7, 10-tetraazacyclododecane- l,4,7-triyl)triacetic acid (DOTA-alkyne).
  • DO A 2,2',2"-(10-(2-oxo-2-(prop-2-yn- l-ylamino)ethyl)- 1,4,7, 10-tetraazacyclododecane- l,4,7-triyl)triacetic acid
  • the chelating compound is optionally derivatized 2-[4,7-bis(carboxymethyl)-l,4,7-triazonan- l- yl] acetic acid (NOT A), such as 2- [4,7-bis(carboxymethyl)- l,4,7-triazonan- l-yl] acetic acid propargylamide (NOTA-alkyne) or optionally derivatized NOTA analogs such as 2-[4,7-bis(carboxymethyl)- l,4,7-triazonan- l-yl]butanoic acid or 2-[4,7- bis(carboxymethyl)- l,4,7-triazonan-l-yl](4-carboxy-4-butanoic acid), and their corresponding propargylamide derivatives.
  • NOT A 2-[4,7-bis(carboxymethyl)-l,4,7-triazonan- l- yl] acetic acid
  • NOTA analogs such as 2-[
  • the chelating compound is optionally derivatized 2,2'-(7-(4-(2-((2-aminoethyl)amino)-2- oxoethyl)benzyl)-l,4,7-triazonane-l,4-diyl)diacetic acid (NOD A) such as 2,2'-(7-(4- (2-oxo-2-(prop-2-yn- l-ylamino)ethyl)benzyl)- 1 ,4,7-triazonane- 1 ,4-diyl)diacetic acid (NODA-alkyne) and related analogs such as 2,2'-(7-(4-(2-oxo-2-(prop-2-yn-l- ylamino)ethyl)benzyl)-l,4,7-triazonane-l,4-diyl)diacetic acid.
  • NOD A such as 2,2'-(7-(
  • the visualization component is a radiographic visualization component.
  • the radiographic visualization component contains 123 I. In some embodiments, the radiographic visualization component contains 99m Tc.
  • the visualization component is a Magnetic Particle Imaging (MPI) visualization component.
  • the MPI component comprises a magnetic nanoparticle.
  • the magnetic nanoparticle has been optimised for MPI.
  • the glycopeptide antibiotic of the construct is selected from the group consisting: of vancomycin; teicoplanin; oritavancin; telavancin; chloroeremomycin; and balhimycin.
  • the glycopeptide antibiotic is vancomycin.
  • the first linker of the construct is at least partially hydrophilic.
  • the first linker comprises a polyethylene glycol (PEG) moiety.
  • PEG polyethylene glycol
  • the PEG moiety is at least PEG3.
  • the PEG moiety is PEG3 or PEG4.
  • the first linker of the construct is hydrophobic.
  • the first linker comprises a linear carbon chain of greater than four carbons.
  • the linear carbon chain is C4-C12.
  • the linear carbon chain is C8.
  • the linear carbon chain is Cl l.
  • the first linker comprises one or more nitrogen- containing moieties.
  • the one or more nitrogen-containing moieties include an amine-derived moiety and/or an azide-derived moiety.
  • the first linker comprises a nitrogen-containing moiety at a first end of the linker.
  • the moiety is an amine-derived moiety.
  • the moiety connects the linker to the glycopeptide antibiotic.
  • the nitrogen-containing moiety may be an amide bond formed between an amine group from a first end of a precursor to the first linker and the C-terminal carboxy moiety from the glycopeptide antibiotic.
  • the first linker comprises a nitrogen-containing moiety at a second end of the linker.
  • the first linker is connected to the visualization component via the nitrogen-containing moiety.
  • this moiety is an azide-derived moiety.
  • this moiety is a triazole moiety.
  • the first linker may be connected to the visualization component via a second linker.
  • the second linker comprises a linear carbon chain.
  • the linear carbon chain is a CI to C4 linear carbon chain.
  • the second linker is connected to the first linker via an azide-derived moiety at the second end of the first linker.
  • the azide-derived moiety is a triazole.
  • the triazole moiety may be formed between an azide group from a second end of a precursor to the first linker and an alkyne group of a precursor to the second linker.
  • the visualization component is preferably connected to the first and/or second linker via a moiety selected from the group consisting of an amide; amine; sulphide; urethane; urea; sulphonamide; ether; or thioester moiety.
  • the moiety is of the second linker.
  • the moiety is an amide moiety.
  • the moiety is an amine moiety.
  • a method of producing a construct including the steps of obtaining (i) an optionally derivatized glycopeptide antibiotic; (ii) a visualization component; and (iii) a first linker, and connecting (i) and (ii) using (iii).
  • the method includes the step of:
  • the construct produced according to the method of this aspect is the construct of the first aspect.
  • a construct comprising (i) an optionally derivatized glycopeptide antibiotic; (ii) a visualization component; and (iii) a first linker connecting (i) and (ii), the method including the steps of:
  • the construct is the construct of the first or second aspect.
  • the microorganism of step (a) is a Gram positive bacteria. In some preferred embodiments the microorganism is a pathogenic microorganism.
  • a fourth aspect there is provided a method of visualizing a microorganism or component thereof, the method including the steps of:
  • the construct of step (a) is the construct of the first or second aspects.
  • the microorganism of step (a) is as described for the third aspect.
  • Visualization according to the method of the fourth aspect may be by any suitable approach.
  • the visualization component of the construct is a fluorescent probe, preferably the visualization is by fluorescence detection.
  • the visualization component of the construct is an MRI visualization component, preferably the visualization is by MRI.
  • the visualization component of the construct is a PET visualization component, preferably the visualization is by PET.
  • the visualization component is an MPI visualization component, preferably visualization is by MPI.
  • the visualization component is a radiographic visualization component, preferably the visualization is by a radiological approach.
  • Visualization according to the method of the fourth aspect may be in vitro visualization, or in vivo visualization.
  • step (a) will include adding the construct to a sample containing the microorganism.
  • the sample of step (a) may be a sample obtained from a biological subject.
  • the subject is a human or an animal.
  • the sample may be urine, sputum, blood or a blood product including platelets, plasma, and/or serum.
  • the sample of step (a) is a blood sample.
  • the blood sample is human blood.
  • the blood sample may comprise aggregated red and/or white blood cells.
  • step (a) will include administering the construct to a biological subject.
  • the subject is a human or an animal.
  • a method of analysing a microorganism or component thereof including the steps of:
  • step (d) analysing the microorganism or component thereof based on the visualization of step (c).
  • steps (a)-(c) are as set forth for the fourth aspect.
  • Analysis of step (d) may be in vitro or in vivo analysis.
  • step (c) is will be in vitro visualization.
  • the visualization of step (c) will be in vivo visualization.
  • the in vivo analysis will be performed in a subject to which the construct has been administered for the visualization of step (c).
  • a method of diagnosing and/or monitoring a disease, disorder or condition including the steps of analysing a microorganism or component thereof according to the method of the fifth aspect, and diagnosing a disease, disorder or condition based on the analysis of the microorganism.
  • the disease, disorder or condition is caused by a Gram positive bacteria.
  • the disease, disorder or condition is a Gram positive bacterial infection.
  • the disease, disorder or condition is sepsis.
  • the disease, disorder or condition is a urinary tract infection.
  • a method of treating a disease, disorder or condition including the steps of diagnosing a disease, disorder or condition according to the method of the sixth aspect, and treating the disease or condition based on the diagnosis.
  • a method of inhibiting, controlling, or killing a microorganism including the step of contacting a construct of the first aspect with a microorganism, to thereby inhibit, control, or kill the microorganism.
  • the step of contacting the construct with the microorganism includes the step of selectively binding the glycopeptide antibiotic of the construct to the microorganism.
  • the microorganism is a Gram positive bacteria.
  • the Gram positive bacteria is an antibiotic resistant bacteria.
  • the microorganism shows at least partial resistance to the glycopeptide antibiotic of the construct when present as a free antibiotic.
  • compositions for diagnosing, treating or preventing a disease, disorder, or condition in a subject comprising a construct of the first aspect, or a construct produced according to the method of the second aspect.
  • a method of diagnosing, treating or preventing a disease, disorder, or condition in a subject including the step of administering to a subject an effective amount of the construct of the first aspect, a construct produced according to the method of the second aspect, or a composition of the ninth aspect, to thereby treat or prevent the disease, disorder, or condition in the subject.
  • the invention provides for use of a construct of the first aspect, or a construct produced according to the method of the second aspect, in the manufacture of a composition for the diagnosis, treatment or prevention of a disease, disorder, or condition in a subject.
  • the disease, disorder, or condition according to the ninth to eleventh aspects is a disease caused by a Gram positive bacteria.
  • the disease is bacterial sepsis.
  • the disease is a urinary tract infection.
  • the invention is directed to a method of increasing the activity or efficacy of a glycopeptide antibiotic, the method including the step of connecting a glycopeptide antibiotic to a visualization component by a first and/or second linker.
  • the glycopeptide antibiotic, the visualization component, and the first and/or second linker are as set forth for the first aspect.
  • a method of assessing a compound for activity in disrupting an outer cell membrane of a Gram negative microorganism including the steps of:
  • construct comprising (i) an optionally derivatized glycopeptide antibiotic; (ii) a visualization component; and (iii) a first linker connecting (i) and (ii); and
  • the degree of binding of the construct to the microorganism is related to the degree of activity of the compound in disrupting the cell membrane of the microorganism.
  • the present invention relates to glycopeptide antibiotic adducts.
  • a first aspect of the second broad form provides a glycopeptide antibiotic adduct comprising an optionally derivatized glycopeptide antibiotic bound to a first linker.
  • Compounds of this broad form will be suitable for connection to a visualization component, to form constructs of the first aspect of the first broad form.
  • the glycopeptide antibiotic of the second broad form is selected from the group consisting of vancomycin; teicoplanin; oritavancin; telavancin; chloroeremomycin; and balhimycin.
  • the glycopeptide antibiotic is vancomycin.
  • the first linker of the second broad form comprises a PEG group, preferably at least PEG3.
  • the first linker comprises a linear carbon chain of greater than four carbons.
  • the linear carbon chain is C4-C12.
  • the linear carbon chain is C8.
  • the linear carbon chain is C 11.
  • the first linker comprises one or more nitrogen- containing compounds.
  • the first linker comprises an amine-derived moiety.
  • the amine-derived moiety is at a first end of the first linker.
  • the amine-derived moiety is an amide bond connecting the linker to the glycopeptide antibiotic.
  • the first linker comprises an azide moiety.
  • the azide moiety is at a second end of the first linker opposite the glycopeptide antibiotic.
  • a method of inhibiting, controlling, or killing a microorganism including the step of contacting an adduct of the first aspect of the second broad form with a microorganism, to thereby inhibit, control, or kill the microorganism.
  • compositions for treating or preventing a disease, disorder, or condition in a subject comprising a glycopeptide antibiotic adduct of the first aspect of the second broad form.
  • a method of treating or preventing a disease, disorder, or condition in a subject including the step of administering to a subject an effective amount of the adduct of the first aspect of the second broad form, or a composition of the third aspect of the second broad form, to thereby treat or prevent the disease, disorder, or condition in the subject.
  • the invention provides for use of an adduct of the first aspect of the second broad form in the manufacture of a composition for the treatment or prevention of a disease, disorder, or condition in a subject.
  • the invention is directed to a method of increasing the activity or efficacy of a glycopeptide antibiotic, the method including the step of connecting a glycopeptide antibiotic to a first linker.
  • the first linker comprises a linear carbon chain.
  • the linear carbon chain is C8.
  • indefinite articles “a” and “an” are not to be read herein as singular indefinite articles or as otherwise excluding more than one or more than a single subject to which the indefinite article refers.
  • an antibiotic includes one antibiotic, one or more antibiotics, or a plurality of antibiotics.
  • Figure 1 sets forth the structure of vancomycin (1) and the glycopeptide antibiotic adducts N 3 -C8-Van (2) and N 3 -PEG3-Van (3).
  • Figure 2 sets forth (A) the synthesis of NBD-alkyne (10) and DMACA-alkyne (11); and (B) the structure of constructs NBD-Tz-C8-Van (12), DMACA-Tz-C8-Van (13), and NBD-Tz-PEG3-Van (14), and a schematic representation of the synthesis of these constructs from N 3 -C8-Van (2) or N 3 -PEG3-Van (3), using NBD-alkyne (10) or DMACA-alkyne (11), respectively.
  • reagents and conditions are (i) propargylamine, HATU, DIPEA, DMF, rt; and (ii) propargylamine, Cs2C03, THF.
  • Figure 3 sets forth vancomycin labelled with keys for NMR elucidation, with reference to the data provided in FIGS. 4 and 7.
  • Figure 4 sets forth NMR data of glycopeptide antibiotic adducts N 3 -C8-Van (2) and N 3 -PEG3-Van (3).
  • Figure 5 sets forth MS/MS data for N 3 -C8-Van (2).
  • Figure 6 sets forth MS/MS data for N 3 -PEG3-Van (3).
  • Figure 7 sets forth NMR data of constructs NBD-Tz-C8-Van (12), DMACA- Tz-C8-Van (13), and NBD-Tz-PEG3-Van (14).
  • Figure 8 sets forth MS/MS data for NBD-Tz-C8-Van (12).
  • Figure 9 sets forth MS/MS data for DMACA-Tz-C8-Van (13).
  • Figure 10 sets forth MS/MS data for NBD-Tz-PEG3-Van (14).
  • Figure 11 sets forth antimicrobial activity (measured in MIC) of glycopeptide antibiotic adducts N 3 -C8-Van, N -C3-Van, and N -PEG3-Van, and constructs NBD- Tz-C8-Van, DMACA-Tz-C8-Van, and NBD-Tz-PEG3-Van against Gram-positive bacteria. Note that there is some overlap in the data presented in the respective charts.
  • Figure 12 sets forth SR-SIM fluorescence imaging of S. aureus (ATCC 25923) stained with (A) Green; NBD-Tz-C8-Van (12), Red; FM4-64FX (bacterial membrane), Blue; Hoechst 33342 (nucleic acid) (B) Blue; DMACA-Tz-C8-Van (13), Red; FM4-64FX (bacterial membrane), Green; SYTO 21 (nucleic acid), (C) Green; NBD-Tz-PEG3-Van (14), Red; FM4-64FX (bacterial membrane), Blue; Hoechst 33342 (nucleic acid).
  • Figure 13 sets forth SR-SIM fluorescence imaging of S. aureus cell division
  • A Green; NBD-Tz-C8-Van (12), Red; FM4-64FX (bacterial membrane), Blue; Hoechst 33342 (nucleic acid)
  • B Blue; DMACA-Tz-C8-Van (13), Red; FM4-64FX (bacterial membrane), Green; SYTO 21 (nucleic acid) (C) Green; NBD-Tz-PEG3- Van (14), Red; FM4-64FX (bacterial membrane), Blue; Hoechst 33342 (nucleic acid).
  • Figure 14 sets forth a cross section of fluorescent imaging of S. aureus (ATCC 25923)
  • A Blue; DMACA-Tz-C8-Van (13), Red; FM4-64FX (bacterial membrane), Green; SYTOX green (nucleic acid);
  • B Combination of DMACA-Tz-C8-Van (13) (Blue) with TMP-NBD probe (Green)
  • C Combination of DMACA-Tz-C8-Van (13) with linezolid-NBD probe (Green). Co-stained with Red; FM4-64FX (bacterial membrane).
  • Figure 15 sets forth structures of HXPI (A), PXPI (B), and DOTA-alkyne (C).
  • Figure 16 sets forth structures of PXPI-Tz-C8-Van (A), PXPI-Tz-PEG3-Van
  • Figure 17 sets forth structures of DOTA-Tz-C8-Van (A) and DOTA-Tz-
  • Figure 18 sets forth the degree of construct binding to E. coli exposed to various compounds.
  • Figure 19 sets forth structures of bisamineoxime, tetraamine, NOD A, and NOT A derivatives.
  • This invention relates to the design and production of constructs comprising a glycopeptide antibiotic and a visualization component.
  • constructs include constructs for microorganism visualization, and constructs with antimicrobial activity.
  • the invention also relates to adducts comprising an optionally derivatized glycopeptide antibiotic bound to a linker, suitable for connection to a visualization component to form such constructs.
  • the invention is at least partly predicated on the recognition that such constructs may offer important advantages for visualization of microorganisms or components thereof in biological samples. Furthermore, the invention is at least partly predicated on the surprising discovery that certain constructs comprising a glycopeptide antibiotic and a visualization component, and/or certain glycopeptide antibiotic adducts, may have substantially increased activity or efficacy as compared to the corresponding free antibiotic itself.
  • the invention is also at least partly predicated on the discovery of design parameters with surprising benefits for binding of Gram positive bacteria using constructs as herein described.
  • Such advantages for binding efficiency may lead to advantages for visualization of Gram positive bacteria using constructs of the invention, and/or for antimicrobial activity of constructs of the invention against Gram positive bacteria.
  • Preferred constructs may have particular advantages with regards to selectivity for binding to bacteria in preference to mammalian cells.
  • Constructs of the invention will comprise an optionally derivatized glycopeptide antibiotic connected to a visualization component.
  • the terms "connect”, “connection”, “connected” etc. will be understood to encompass direct connection (e.g. direct binding or direct linkage), or indirect connection (e.g. connection via one or more other molecules).
  • the construct of this aspect is for binding to a microorganism or component thereof, wherein the optionally derivatized glycopeptide antibiotic binds to the microorganism or component thereof.
  • One aspect of the invention relates to a construct comprising: (i) an optionally derivatized glycopeptide antibiotic; (ii) a visualization component; and (iii) a first linker connecting (i) and (ii).
  • a "derivatized" glycopeptide antibiotic broadly encompasses glycopeptide antibiotics comprising one or more modifications or alterations such as transformations of existing functional groups and introduction of temporary protecting groups and the like. The term is also considered to include all salt forms.
  • the derivatized glycopeptide antibiotic will be a biologically active derivative, which retains at least a part of one or more biological activities of a corresponding non- derivatized or unmodified glycopeptide antibiotic.
  • Biological activities of a glycopeptide antibiotic may include Gram positive bacteria and/or antimicrobial activity; and/or peptidoglycan and/or Lipid II binding, although without limitation thereto.
  • the derivatized glycopeptide antibiotic retains at least: 10%; 20%; 30%; 40%; 50%; 60; 70%; 80%; or 90% of one or more biological activities.
  • modifications of glycopeptide antibiotics that may result in biologically active derivatives, the skilled person is directed to Malabarba et al. (1997) Medicinal Research Reviews, 17(1) 69-137, incorporated herein by reference.
  • Constructs of this aspect may have the following general structure:
  • A is the optionally derivatized glycopeptide antibiotic
  • Li is the first linker
  • V is the visualization component.
  • connections between A and Li; and Li and V may be direct or indirect connections.
  • the connection between A and Li is a direct connection.
  • the connection between Li and V is an indirect connection.
  • the construct comprises:
  • A is the optionally derivatized glycopeptide antibiotic
  • Li is the first linker
  • L 2 is the second linker
  • V is the visualization component.
  • connections between A and Li; Li and L 2 ; and L 2 and V may be direct or indirect connections.
  • the connection between A and Li is a direct connection.
  • the connection between and Li and L 2 is a direct connection.
  • the connection between L 2 and V is a direct connection.
  • construct of this aspect may be depicted as follows:
  • A is the optionally derivatized glycopeptide antibiotic
  • W is a functionality of the first linker (Li) linking Li to A;
  • X is a functionality of Li linking Li to the second linker (L 2 );
  • Y is a functionality of L 2 linking L 2 to V;
  • Y is the visualization component.
  • W comprises an amide bond
  • X comprises a triazole
  • Y comprises an amide, amine, sulphide, or thioester, urethane, urea, sulphonamide, or ether moiety
  • constructs of the invention may be depicted as follows:
  • A is the optionally derivatized glycopeptide antibiotic
  • W-Ri-X is the first linker comprising functional groups W and X on first and second ends, respectively, and an internal moiety Ri;
  • R 2 -Y is the second linker comprising moiety R 2 and functional group Y;
  • V is the visualization component.
  • W is an amide bond connecting the first linker to A
  • Ri comprises a PEGN moiety (as hereinbelow described) and/or a linear carbon chain;
  • X is a triazole connecting the first linker and R 2 of the second linker
  • R 2 comprises a linear carbon chain of at least C 1 ;
  • V is an amine, amide, or sulphonamide bond connecting the second linker to
  • W-PEG3-X is the first linker comprising W (amide bond) and X (triazole) on first and second ends respectively, and an internal PEG3 moiety.
  • Cl-Y is the second linker comprising a CI chain and Y (amine or amide bond);
  • V is a fluorescent visualization component selected from the group consisting of optionally derivatized: NBD; DMACA; dansyl and BODIPY.
  • FIG. 2 Exemplary embodiments of constructs in the form A-W-PEG3-X-C1-Y-V are set forth in FIG. 2.
  • constructs of the invention are designed for binding to a microorganism or a component thereof to facilitate visualization or imaging of the microorganism or component thereof.
  • the terms “visualization”, “visualize”, and “visualized” etc., and “imaging”, “image”, and “imaged” etc. refer to the making of a visual representation.
  • binding of preferred constructs to a microorganism or component thereof will provide some suitable signal or tag for visual representation.
  • the visualization may be direct, i.e. facilitated by an emission of light in the visual spectrum from the construct.
  • the visualization may also be indirect, i.e. facilitated by emission of a signal from the construct that can be converted into a visual representation by suitable processing.
  • Preferred constructs of the invention are particularly suited to visualization of Gram positive bacteria.
  • visualization of other microorganisms which bind to glycopeptide antibiotics is also within the scope of the invention.
  • certain glycopeptide antibiotics have been shown to inhibit fungal growth, although the mechanism of inhibition (and in particular, whether this involves binding to fungal components) is presently unclear in at least many instances.
  • preferred constructs of this aspect bind Lipid II and/or peptidoglycan, wherein the optionally derivatized glycopeptide antibiotic binds to the Lipid II and/or peptidoglycan.
  • constructs of this aspect that bind Lipid II and/or peptidoglycan will typically be suitable for visualization of Gram positive microorganisms as set forth above, it will be readily appreciated that such constructs will also typically be suited to visualization of Lipid II and/or peptidoglycan itself, regardless of origin, provided that the optionally derivatized glycopeptide antibiotic of the construct can access and bind the Lipid II and/or peptidoglycan.
  • the Lipid II and/or peptidoglycan may be contained within any suitable microorganism. It will be further appreciated that the Lipid II and/or peptidoglycan may be visualized after being obtained or produced by any suitable means, such as extraction from a microorganism, or by synthetic production.
  • construct of this aspect may have application in visualization of Gram negative bacteria in certain circumstances.
  • visualization constructs of this aspect can be desirable for the assessment of outer membrane damage to Gram negative bacteria.
  • cell walls of Gram negative bacteria also comprise peptidoglycan, however this is located underneath an outer membrane, and typically not accessible to binding by antibiotics while the outer membrane is intact.
  • constructs of the invention can be applied to the assessment of membrane damage in Gram negative bacteria, wherein binding of the construct the Gram negative bacteria indicates that the Gram negative bacteria has incurred membrane damage, and peptidoglycan in the cell wall has been exposed.
  • visualization using constructs of the invention is in vitro visualization.
  • in vitro visualization will be understood to refer to any visualization that occurs outside of a biological subject.
  • In vitro visualization may be performed using any suitable sample containing a suitable microorganism or component thereof, and optionally other sample components.
  • samples may include laboratory samples, such as artificial cultures; and environmental samples such as soil and water samples.
  • samples may also include samples obtained or removed from a biological subject, including samples from plants and animals.
  • the sample is an animal sample.
  • the sample may be from a human; a primate (e.g. apes and monkeys); a canine; a feline; an ungulate (e.g. equine, bovine, and swine); or an avian, although without limitation thereto.
  • the animal may be livestock (e.g. horses, cattle and sheep), a companion animals (e.g. dogs and cats), a laboratory animals (e.g. mice, rats and guinea pigs) or a performance animal (e.g. racehorses, greyhounds and camels), although without limitation thereto.
  • the animal sample may be of a mammalian or non-mammalian species.
  • the animal sample is a human sample.
  • visualization using constructs of the invention is in vivo visualization.
  • in vivo visualization will be understood to refer to any visualization that occurs within a biological subject.
  • in vivo visualization will exclude analysis performed within a sample obtained or removed from a biological subject, which will be considered to be in vitro visualization, as hereinabove described.
  • in vivo visualization is performed within an animal.
  • the animal may be a human; a primate (e.g. apes and monkeys); a canine; a feline; an ungulate (e.g. equine, bovine, and swine); or an avian, although without limitation thereto.
  • the animal may be livestock (e.g. horses, cattle and sheep), a companion animals (e.g. dogs and cats), a laboratory animals (e.g. mice, rats and guinea pigs) or a performance animal (e.g. racehorses, greyhounds and camels), although without limitation thereto.
  • the animal sample may be of a mammalian or non-mammalian species.
  • in vivo visualization is performed within a human.
  • glycopeptide antibiotic will be understood to be a glycosylated peptide with at least some capacity to (i) bind and/or (ii) inhibit growth, proliferation or viability of a microorganism.
  • antimicrobiaF activity the ability to inhibit growth, proliferation and/or viability of a microorganism may be referred to generally as " antimicrobiaF activity.
  • glycopeptide antibiotics comprise cyclic or polycyclic peptides.
  • Glycopeptide antibiotics typically have properties consistent with microbial, nonribosomal origin. However, it will be appreciated that all suitable compounds regardless of origin, e.g. both isolated natural compounds and synthetically produced or recombinant compounds, are encompassed by the term glycopeptide antibiotic as used herein.
  • isolated is meant material that has been removed from its natural state or otherwise been subjected to human manipulation. Isolated material may be substantially or essentially free from components that normally accompany it in its natural state, or may be manipulated so as to be in an artificial state together with components that normally accompany it in its natural state.
  • Glycopeptide antibiotics of constructs of the invention preferably have capacity to bind Gram positive bacteria. It will be appreciated by the skilled person that glycopeptide antibiotics typically bind to peptidoglycan in the bacterial cell wall and to the Lipid II precursor of peptidoglycan. Typically, glycopeptide antibiotics bind selectively to Gram positive bacteria, with limited or absent binding activity towards Gram negative bacteria.
  • the glycopeptide antibiotic of the construct may be any suitable glycopeptide antibiotic.
  • the glycopeptide antibiotic is selected from the group consisting: of vancomycin; teicoplanin; oritavancin; telavancin; dalbavancin; chloroeremomycin; and balhimycin.
  • the glycopeptide antibiotic is vancomycin. It will be appreciated that, typically, binding of peptidoglycan of the cell wall of Gram positive bacteria by glycopeptide antibiotics occurs in a substantially non- species or strain -specific manner. That is, glycopeptide antibiotics will generally have broad binding activity for Gram positive bacteria as a group. It will be further understood that glycopeptide antibiotics will generally bind to Gram positive bacteria regardless of whether the bacteria is sensitive or resistant to the antibiotic.
  • binding of the glycopeptide antibiotic may or may not inhibit growth, proliferation, or viability of the bound microorganism. It will be appreciated that, in some embodiments, the viability of a microorganism bound using the construct may be of limited or no significance for the desired visualization application. For other applications, viable or non-viable microorganisms may be preferred. Accordingly, for these embodiments, a suitable antibiotic that is optimized to either maintain or disrupt viability of a microorganism of interest may be selected.
  • constructs according to certain embodiments of the invention are designed to inhibit or kill microorganisms to which the glycopeptide antibiotic of the construct binds. Accordingly, for these embodiments, a suitable antibiotic that is optimized to have at least partial antimicrobial activity against a microorganism of interest may be selected.
  • a "visualization component” broadly includes any entity with the capacity to be connected to a glycopeptide antibiotic to form a construct of the invention, and to facilitate visualization or imaging using the construct.
  • visualization components of constructs of the invention will be designed to facilitate imaging of microorganisms or components thereof bound to the constructs.
  • Visualization components of the construct of the invention may take a range of suitable forms.
  • the visualization component is a light- emitting component.
  • Light-emitting components broadly include luminescent and incandescent sources of light.
  • the light emitting component is a luminescent component.
  • the light emitting component comprises a fluorescent compound.
  • the light emitting component comprises a quantum dot.
  • the light emitting component emits light in the visual spectrum, i.e. in a wavelength range of about 400 nm to about 700 nm.
  • Visualization components comprising fluorescent compounds emitting light in the visual spectrum may be referred to alternatively herein as "fluorescent probes ' " .
  • the light emitting component emits light in the infrared spectrum, i.e. in a wavelength range of about 700 nm to about 1mm.
  • the infrared spectrum is a near infrared spectrum.
  • the “near infrared” spectrum will be understood to include the wavelength range of about 700 nm to about 1500 nm.
  • the infrared spectrum is a mid infrared spectrum.
  • the “mid infrared' spectrum will be understood to include the wavelength range of about 1500 nm to about 5 ⁇ .
  • the infrared spectrum is a far infrared spectrum.
  • the "far infrared' spectrum will be understood to include the wavelength range of about 5 ⁇ to about 1 mm.
  • infrared probes Visualization components comprising fluorescent compounds emitting light in the infrared spectrum may be referred to alternatively herein as "infrared probes".
  • the fluorescent compound has a neutral charge.
  • the fluorescent compound has a molecular weight between about 100 and about 500 Da.
  • the molecular weight is between about 160 and about 250 Da.
  • the fluorescent compound is an organic or carbon- containing compound.
  • the organic fluorescent compound comprises one or more heterocyclic groups.
  • the heterocyclic groups are N or O -containing heterocyclic groups.
  • the heterocyclic group is or comprises a benzofurazan group. In one preferred embodiment, the heterocyclic group is or comprises a coumarin group. In one preferred embodiment, the heterocyclic group is or comprises a naphthalene group. In one preferred embodiment, the heterocyclic group is or comprises an isochromene group.
  • the fluorescent compound is optionally derivatized 7-nitrobenz-2-oxa-l,3-diazol-4-yl (NBD). In one particularly preferred embodiment, the fluorescent compound is optionally derivatized 7- (dimethylamino)-coumarin-4-acetic acid (DMACA).
  • the fluorescent compound is optionally derivatized 5-(dimethylamino)-l naphthalene- 1-sulfonyl (dansyl). In one particularly preferred embodiment, the fluorescent compound is optionally derivatized 4,4-difluoro-4-bora-3a,4a-diaza-s- indacene (BODIPY).
  • the visualization component is a Magnetic Resonance Imaging (MRI) visualization component.
  • MRI Magnetic Resonance Imaging
  • Suitable components to facilitate visualization by MRI have been described in the art. The skilled person is directed to Pierre et al (2014) Journal of Biological and Inorganic Chemistry, 19 127-131 (incorporated herein by reference) in this regard.
  • the MRI visualization component comprises 19 F.
  • the MRI visualization component comprises a gadolinium complex.
  • the gadolinium is Gd(III).
  • the MRI visualization component comprises a Europium complex.
  • the Europium is Eu(II).
  • the MRI visualization component comprises diethylenetriamine penta-acetic acid (DTPA), such as Gd-DTPA or Eu-DTPA.
  • DTPA diethylenetriamine penta-acetic acid
  • the MRI visualization component comprises a chemical exchange transfer (CEST) agent.
  • the MRI visualization component comprises a paramagnetic chemical exchange saturation transfer (PARACEST) agent.
  • Visualization components that are MRI visualization components may be referred to alternatively herein as "MRI probes" .
  • the visualization component is a Positron Emission
  • PET Tomography
  • the PET visualization component comprises a 64 Cu complex. In some embodiment, the PET visualization component comprises a 68 Ga complex. In some embodiments, the PET visualization component comprises U C. In some embodiments, the PET visualization component comprises 18 F. In some embodiments, the PET visualization component comprises 124 I. In some embodiment, the PET visualization component contains 99m Tc.
  • the PET visualization component may comprise a chelating compound, such as part of a metal complex. Structures of some suitable chelating compounds for PET applications are set forth in FIG. 19.
  • the chelating compound is optionally derivatized l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (DOTA), such as 2,2',2"-(10-(2-oxo-2-(prop-2-yn-l-ylamino)ethyl)- 1,4,7, 10-tetraazacyclododecane- l,4,7-triyl)triacetic acid (DOTA-alkyne).
  • DOTA 2,2',2"-(10-(2-oxo-2-(prop-2-yn-l-ylamino)ethyl)- 1,4,7, 10-tetraazacyclododecane- l,4,7-triyl)triacetic acid
  • the chelating compound is optionally derivatized 2- [4,7-bis(carboxymethyl)-l,4,7-triazonan-l-yl] acetic acid (NOTA), such as 2-[4,7-bis(carboxymethyl)-l,4,7-triazonan-l-yl]acetic acid propargylamide (NOTA-alkyne) or optionally derivatized NOTA analogs such as 2-[4,7- bis(carboxymethyl)-l,4,7-triazonan-l-yl]butanoic acid or 2-[4,7-bis(carboxymethyl)- l,4,7-triazonan-l-yl](4-carboxy-4-butanoic acid), and their corresponding propargylamide derivatives.
  • NOTA 2- [4,7-bis(carboxymethyl)-l,4,7-triazonan-l-yl] acetic acid
  • NOTA analogs such as 2-[4,7- bis(carboxymethyl)-
  • the chelating compound is optionally derivatized 2,2'-(7-(4-(2-((2-aminoethyl)amino)-2-oxoethyl)benzyl)-l,4,7-triazonane- l,4-diyl)diacetic acid (NODA), such as 2,2'-(7-(4-(2-oxo-2-(prop-2-yn-l- ylamino)ethyl)benzyl)-l,4,7-triazonane-l,4-diyl)diacetic acid (NODA-alkyne) and related analogs such as 2,2'-(7-(4-(2-oxo-2-(prop-2-yn-l-ylamino)ethyl)benzyl)- 1,4,7- triazonane- 1 ,4-diyl)diacetic acid.
  • NODA 2,2'-(7-(4-(2-((2-
  • the visualization component is a Single photon emission computed tomography (SPECT) visualization component.
  • SPECT Single photon emission computed tomography
  • the SPECT visualization component may comprise a chelating compound, such as part of a metal complex.
  • the chelating compound is optionally derivatized bisamineoxime (FIG. 19).
  • the chelating compound is optionally derivatized tetraamine (FIG. 19).
  • PET probes PET or SPECT visualization components
  • SPECT probes SPECT probes
  • the visualization component is a radiographic visualization component.
  • the radiographic visualization component contains 123 I.
  • the radiographic visualization component contains 99m Tc.
  • Visualization components that are radiographic visualization components may be referred to alternatively herein as "radiographic probes”.
  • the visualization component is a Magnetic Particle Imaging
  • MPI visualization component. Suitable components to facilitate visualization by MPI have been described in the art. The skilled person is directed to Panagiotopoulos et al (2015) International Journal of Nanomedicine, 10 3097-3114 (incorporated herein by reference) in this regard.
  • the MPI visualization component is preferably a magnetic nanoparticle.
  • the magnetic nanoparticle is a superparamagnetic nanoparticle.
  • the MPI visualization component is superparamagnetic iron oxide nanoparticle (SPION).
  • SPION superparamagnetic iron oxide nanoparticle
  • Visualization components that are MPI visualization components may be referred to alternatively herein as "MPI probes”.
  • Linkers of constructs of the invention facilitate connection between the glycopeptide antibiotic and the visualization component.
  • constructs comprising certain linkers are advantageous for binding to microorganisms such as Gram positive bacteria, to facilitate visualization.
  • Certain linkers which are at least partially hydrophilic may be particularly effective for binding and visualization using constructs of the invention.
  • molecules which are at least partially hydrophilic may be particularly effective for binding to microorganisms to facilitate visualization using constructs of the invention.
  • partially hydrophilic or hydrophilic linkers e.g. those comprising PEG as herein described, may provide an extended tether for the glycopeptide antibiotic, with limited or absent coiling occurring in an aqueous environment.
  • the linker comprises a polyethylene glycol (PEG) group.
  • PEG molecules may be expressed, when located internally within a larger molecule as in constructs of the invention, in the form R a -(0-CH 2 -CH 2 )n- where 'n' is the number of PEG monomers and R a is a carbon chain connecting the PEG.
  • PEG molecules may be expressed in the form 'PEGN', wherein 'N' is the number of PEG monomers.
  • R a comprises a C2-C4, or more preferably C2 linear carbon chain.
  • PEG-containing moieties may take the form -(CH 2 ) m (0- CH 2 -CH 2 ) consult- where 'm' is 2 to 4 carbons e.g. -CH 2 -CH 2 -(0-CH 2 -CH 2 ) 3 -. It will be appreciated that a determination of the particular side or end of the PEG moiety to which a linear carbon chain, such as a C2 chain, is adjacent, may be dependent upon the end of the combined PEG-linear carbon chain molecule that is considered to be the 'start', or 'first end' of the PEG-linear carbon chain molecule.
  • the PEG group is at least PEG3.
  • the PEG group is in the range PEG3 to PEG10, including PEG4; PEG5; PEG6; PEG7; PEG8; and PEG9.
  • the PEG group is PEG3 or PEG4.
  • partially hydrophilic first linkers may comprise other components.
  • the partially hydrophilic first linker may comprise a hydrophobic component.
  • the hydrophobic component may be a linear carbon chain.
  • the linear carbon chain may be a C2-C15 chain, including C3; C4; C5; C6; C7; C8; C9; CIO; Cl l; C12; C13; and C14, although without limitation thereto.
  • hydrophobic linkers may also be suitable according to constructs of the invention. It has been recognised that, where hydrophobic linkers are used, molecules comprising at least a particular linear chain length may be more effective for capture using constructs of the invention.
  • the linker is a hydrophobic molecule comprising a linear carbon chain of at least four carbons.
  • the linear carbon chain is C4- C20, including C5; C6; C7; C8; C9; CIO; Cl l; C12; C13; C14; C15; C16; C17; C18; and C19, or preferably C6-C10.
  • the linear carbon chain may be C6-C16; C6-C14; C8- C14; C8-12; C8-10; or CIO- 12. In one particularly preferred embodiment, the linear carbon chain is C8. In another particularly preferred embodiment the linear carbon chain is CI 1. It will be appreciated that the linear carbon chain may be an alkane, an alkene, or an alkyne, although without limitation thereto.
  • the first linker is directly connected or directly bound to the glycopeptide antibiotic (see, e.g. FIGS. 1-2).
  • the first linker may be connected to the glycopeptide antibiotic via reaction with a glycopeptide moiety selected from the group consisting of: a C-terminal carboxy moiety; a primary or secondary N-terminal moiety; a hydroxyl moiety; a phenolic moiety; and an amine moiety.
  • a glycopeptide moiety selected from the group consisting of: a C-terminal carboxy moiety; a primary or secondary N-terminal moiety; a hydroxyl moiety; a phenolic moiety; and an amine moiety.
  • the first linker is connected to the glycopeptide antibiotic via an amide group formed from the C-terminal carboxy moiety.
  • the first linker comprises one or more nitrogen- containing moieties.
  • the nitrogen-containing moieties may be any suitable moieties.
  • the nitrogen-containing moieties may contain one or a plurality of nitrogens. In some preferred embodiments, the nitrogen-containing moieties contain between 1 and 6 nitrogens, or preferably between 1 and 3 nitrogens.
  • the nitrogen-containing moieties may be linear or cyclic moieties, including heterocyclic moieties.
  • the one or more nitrogen-containing moieties include an amine-derived moiety and/or an azide-derived moiety.
  • amine - derived moiety and azide-derived moiety it is intended that, whatever the actual functionality of the relevant connecting bond or moiety in the construct, it was at least in part derived by the coming together of an amine or an azide with a complimentary reactive functional group.
  • an amide bond in the construct could be termed an amine-derived moiety as it may have been formed from reaction of an amine with a carboxy group.
  • the terms cover situations where the amine or other relevant group was present on precursors to the linker, glycopeptide antibiotic or visualization component prior to reaction to form the final construct.
  • the first linker comprises a nitrogen-containing moiety at a first end of the linker.
  • the moiety is an amine-derived moiety.
  • the moiety connects the linker to the glycope tide antibiotic.
  • the connection is a direct connection.
  • the amine-derived moiety will be derived from an amine upon binding to another component forming part of the construct. It will be further understood that the particular identity of the amine-derived moiety will typically depend upon the component of the construct to which the amine-derived moiety is bound.
  • the amine-derived moiety is bound to the glycopeptide antibiotic by the C-terminal carboxy moiety of the glycopeptide antibiotic, the amine-derived moiety will be an amide (see, e.g. FIG. 2).
  • the amine-derived moiety is bound to the glycopeptide antibiotic by a primary or secondary amine group the amine-derived moiety will be a urea.
  • the amine-derived moiety is bound to the glycopeptide antibiotic by a hydroxyl or phenolic group, the amine-derived moiety will be a urethane.
  • the first linker may be connected to the visualization component via a second linker.
  • the second linker comprises a linear carbon chain.
  • the linear carbon chain is C1-C10, including C2; C3; C4; C5; C6; C7; C8; and C9. It will be understood that, as used in this context, a single internal carbon group will be considered a 'linear CI carbon chain' .
  • the linear carbon chain is CI to C4.
  • the linear carbon chain is CI (see, e.g., FIG. 2).
  • the second linker is connected to the first linker via an azide - derived moiety at the second end of the first linker.
  • the azide-derived moiety is a triazole.
  • the triazole moiety may be formed between an azide group from the second end of a precursor to the first linker and an alkyne group of a precursor to the second linker (see, e.g. FIG. 2).
  • the triazole moiety may be alternatively formed, e.g. between an alkyne group from the second end of the first linker and an azide group of a precursor to the second linker.
  • the formation of the triazole from the precursors of the first and second linkers is achievable by rapid or 'click' reaction chemistry (see, e.g., the Examples).
  • click chemistry including in the context of triazole formation, the skilled person is directed to Kolb et al (2003) Drug Discovery Today, 8(24) 1128-1137.
  • the formation of the triazole from the precursors of the first and second linkers as set forth in the Examples involves the use of copper (Cu) catalysis
  • Cu copper
  • the use of an alkyne group within certain moieties, e.g. a strained cyclooctyne can be particularly favourable for formation of a triazole even in the absence of Cu catalysis, such as under biological conditions.
  • the visualization component is preferably connected to the first and/or second linker via a moiety selected from the group consisting of an amide, amine, sulphide, urethane, urea, ether, or thioester moiety. It will be understood that such moieties may be formed by reaction of an amine, hydroxyl or thiol group that was present on the second end of a precursor to the first or second linker, with a group (e.g. a leaving group or carboxyl group) present on a precursor to the visualisation component as present in the construct.
  • a group e.g. a leaving group or carboxyl group
  • the visualization component is connected to a moiety of the second linker.
  • the moiety is an amide moiety (see, e.g., FIG. 2).
  • the moiety is an amine moiety (see, e.g. FIG. 2).
  • a related aspect of the invention is directed to a method of producing a construct, the method including the steps of obtaining (i) an optionally derivatized glycopeptide antibiotic; (ii) a visualization component; and (iii) a first linker, and connecting (i) and (ii) using (iii).
  • the method includes the step of:
  • the construct produced according to the method of this aspect is the construct of the first aspect.
  • the method of this aspect may be or involve in vitro synthesis, such as described in the Examples.
  • a component of the construct comprising the optionally derivatized glycopeptide antibiotic connected to a precursor to the first linker; and a component of the construct comprising the visualization component connected to a precursor to the second linker may be administered separately in vivo, wherein these respective components connect to form the final construct in vivo.
  • the structure of the respective ends of the first and second linkers will be such that connection (e.g. by way of formation of a triazole group as herein described) occurs by click or rapid chemistry, under biological conditions.
  • use of an alkyne group of a strained cyclooctyne for the formation of a triazole connecting the first and second linker may be particular desirable for such embodiments.
  • the following methods encompass embodiments wherein in vivo production of the construct is performed.
  • the microorganisms may be bound to the glycopeptide antibiotic before or after connection of components of the construct to form the final construct in vivo.
  • the invention further provides a method of binding a construct to a microorganism or component thereof, the construct comprising (i) an optionally derivatized glycopeptide antibiotic; (ii) a visualization component; and (iii) a first linker connecting (i) and (ii), the method including the steps of:
  • the construct is a construct as hereinabove described.
  • the microorganism is a Gram positive microorganism.
  • the microorganism component is peptidoglycan and/or Lipid II. It will be appreciated that, in this context, any peptidoglycan and/or Lipid II will be considered a 'microorganism component' . That is, although peptidoglycan and/or Lipid II will typically be derived from a suitable microorganism, peptidoglycan and/or Lipid II obtained by any other suitable means (e.g. synthetically) also falls within the scope of this aspect.
  • step (b) of the method of this aspect by “selectively binding” or “selectively bound” etc., is meant that the microorganism or component thereof is bound with at least partial specificity as compared to other sample components and/or microorganisms.
  • glycopeptide antibiotics typically bind to Gram positive bacteria as a group selectively (as compared, for example, to Gram negative bacteria, or non-Gram staining bacteria).
  • “selectively binding” etc. will refer to binding to Gram positive bacteria as a group with at least partial specificity (rather than binding to a particular Gram positive bacterial species or strain).
  • the construct and the microorganism may be combined in vitro or in vivo, as hereinabove described.
  • the microorganism bound according to step (b) of this aspect is a pathogenic Gram positive bacteria.
  • the pathogenic Gram positive pathogenic bacteria is selected from the group consisting of: Bacillus; Clostridium; Corynebacterium; Enterococcus; Listeria; Staphylococcus; and Streptococcus.
  • the Gram positive bacteria is a cocci e.g.
  • the Staphylococcus is S. aureus.
  • the S. aureus may be, without limitation thereto, a glycopeptide-sensitive strain (such as ATCC 25923); an MRSA stain (such as ATCC 43300); a glycopeptide-intermediate
  • GISA vancomycin-resistant (VRSA) strain
  • VRSA vancomycin-resistant
  • the S. aureus may also be an NARSA VRS 10 strain; a NARSA
  • VRS 3b strain or an NRS 1 strain.
  • the Staphylococcus is S. epidermis.
  • the S. epidermis may be, without limitation thereto, a glycopeptide-sensitive strain (such as ATCC
  • glycopeptide-intermediate (GISE) strain such as NARSA NRS 60.
  • Streptococcus is S. pneumoniae.
  • the S. pneumoniae may be, without limitation thereto, a glycopeptide sensitive strain
  • the Enterococcus is E. faecium.
  • the E. faecium may be, without limitation thereto, a vancomycin A (Van A) resistant strain (such as ATCC 51559) or a vancomycin B (Van B) resistant strain (such as ATCC 51299).
  • the Enterococcus is E. faecalis.
  • the E. faecalis may be, without limitation, a vancomycin sensitive strain (such as ATCC29212) or a vancomycin resistant strain.
  • Another aspect of the invention relates to a method of visualizing a microorganism or component thereof.
  • the method will include the steps of:
  • steps (a) and (b) are as described above for methods of binding a microorganism.
  • Visualization according to step (c) of the method of this aspect may be by any suitable approach.
  • the visualization component of the construct is a fluorescent probe or infrared probe, preferably the visualization is by fluorescence detection.
  • the visualization component is a fluorescent probe
  • the visualization is by Super-Resolution Structured Illumination Microscopy (SR-SEVI) or another high resolution fluorescent microscopy technique.
  • SR-SEVI Super-Resolution Structured Illumination Microscopy
  • the visualization is by Fluorescence Activated Cell Sorter (FACS) technique.
  • FACS Fluorescence Activated Cell Sorter
  • the visualization component of the construct is an MRI component or MRI probe
  • the visualization is by MRI.
  • Techniques for MRI are known in the art.
  • MRI approaches using probes to visualize biological targets such as microorganisms
  • the skilled person is directed to Pierre et al, supra, and Sosnovik et al (2007) Current Opinion in Biotechnology, 18(1) 4-10 (incorporated herein by reference).
  • the visualization component of the construct is PET component
  • the visualization is by PET.
  • Techniques for PET are known in the art; the skilled person is directed to Positron Emission Tomography: Basic Sciences (Springer- Verlag, 2009, Bailey, Townsend, Valk, and Maisey Eds.), incorporated herein by reference.
  • PET approaches using probes to visualize biological targets such as microorganisms
  • the skilled person is directed, by way of example, to Gowrishankar et al (2014) PLOS ONE, 9(9) el07951 (incorporated herein by reference).
  • visualization is by MPI.
  • MPI a technique for MPI are known in the art. The skilled person is directed Panagiotopoulos et al, supra, and Magnetic Particle Imaging (Elsevier, 2010, Kevin R. Minard), incorporated herein by reference, in this regard.
  • the visualization component is a radiographic visualization component
  • the visualization is radiological visualization.
  • Techniques for radiological visualization are known in the art. In particular regard to approaches using radiographic probes for visualization of biological components, the skilled person is directed to Signore et al (2014) Current Pharmaceutical Design, 20(14) 2338-2345 (incorporated herein by reference).
  • the visualization using a radiographic visualization component may be by conventional nuclear medicine planar imaging or SPECT (Single-photon emission computed tomography), although without limitation thereto.
  • visualization according to the method of this aspect may be in vitro visualization, or in vivo visualization.
  • step (a) will include adding the construct to a sample containing the microorganism or component thereof.
  • the sample of step (a) is a sample obtained from a biological subject.
  • the subject is an animal, as hereinabove described.
  • the animal is a human.
  • the biological sample may take any suitable form.
  • the animal or human sample may be a tissue sample including a muscle; epithelial; connective; or nervous tissue sample.
  • the sample may further be a waste product such as urine, or an excretion such as sputum.
  • the sample is a blood sample.
  • the blood sample is human blood.
  • step (a) will include administering the construct to a biological subject.
  • the subject is a human or an animal, as hereinabove described.
  • the construct may be administered to the biological subject in the form of a pharmaceutically acceptable composition as hereinbelow described.
  • Any safe route of administration may be employed for providing a patient with constructs or compositions of the invention for in vivo visualization.
  • enteral, oral, rectal, parenteral, sublingual, buccal, intravenous, intra- articular, intramuscular, intra-dermal, subcutaneous, inhalational, intraocular, intraperitoneal, intracerebroventricular, transdermal and the like may be employed.
  • the administration occurs directly to a site within the body wherein the in vivo visualization is to occur. Additionally or alternatively, the administration may occur to a site within the body outside of where the in vivo visualization is to occur. In these embodiments, the construct will be transported to the site wherein the in vivo visualization is to occur, typically by bodily processes (e.g. blood flow; lymph flow; digestive movement etc.).
  • bodily processes e.g. blood flow; lymph flow; digestive movement etc.
  • the construct is administered to the blood.
  • the visualization may occur in any part of the body of the human or animal.
  • the in vivo visualization may occur within the respiratory system (e.g. the lungs); the digestive system (e.g. the stomach or intestines); the nervous system (e.g. the spine or brain); or the endocrine system (e.g. the pancreas).
  • the in vivo visualization occurs in the circulatory system.
  • the in vivo visualization may occur within one or more tissue types of the biological subject.
  • the visualization may occur in muscle; epithelial; connective; or nervous tissue, although without limitation thereto.
  • in vivo visualization occurs in blood.
  • the invention further provides a method of analysing a microorganism or component thereof.
  • the method will include the steps of:
  • step (d) analysing the microorganism or component thereof based on the visualization of step (c).
  • steps (a)-(c) are as set forth for the directly preceding aspect directed to methods of visualizing a microorganism or component thereof.
  • Analysis according to step (d) may be any suitable analysis. It will be appreciated that the analysis as per step (d) may be an in vitro analysis or an in vivo analysis.
  • microorganism cellular dynamics e.g. bacterial cell and/or cell wall division
  • step (d) may be in vitro or in vivo analysis.
  • the visualization of step (c) is will be in vitro visualization.
  • the in vitro analysis is of a sample obtained from a biological subject.
  • the sample from the biological subject is a human or animal subject as described for the directly preceding aspect.
  • the visualization of step (c) will be in vivo visualization.
  • the in vivo analysis will be performed in a subject to which the construct has been administered for the visualization of step (c).
  • administration for the visualization of step (c) is as described for the directly preceding aspect.
  • the visualization will occur within the body of a human or animal subject as described for the directly preceding aspect.
  • the invention provides for a method of assessing a compound for activity in disrupting a cell membrane of a microorganism.
  • the method will include the steps of:
  • construct comprising (i) an optionally derivatized glycopeptide antibiotic; (ii) a visualization component; and (iii) a first linker connecting (i) and (ii); and (c) determining if the construct is bound to the microorganism by visualization of the construct, wherein
  • the degree of binding of the construct to the microorganism is related to the activity of the construct in disrupting the cell membrane of the microorganism.
  • said degree of binding is positively related to said activity of the compound.
  • the method of this aspect is particularly adapted for screening of compounds for activity in disrupting outer cell membranes of Gram negative microorganisms.
  • Gram negative microorganisms have a cell wall within which peptidoglycan is covered by an outer cell membrane.
  • peptidoglycan-binding antibiotics such as those of constructs described herein do not substantially bind to intact Gram negative microorganisms with intact out cell membranes.
  • disruption of the outer cell membrane such as by exposure of the microorganism to a compound with relevant activity, can result in peptidoglycan of the Gram negative cell wall becoming accessible for binding. Accordingly, it will be readily understood by the skilled person that visualization of a Gram negative microorganism exposed to a compound according to the method of this aspect is indicative of disruption of the outer cell membrane by activity of the compound.
  • the microorganism according to the method of this aspect is E. coli.
  • Another aspect of the invention is directed to a method of diagnosing and/or monitoring a disease or condition.
  • the method will include the steps of analysing a microorganism or component thereof in a sample of a subject, as described for the directly preceding aspect, and diagnosing and/or monitoring a disease or condition based on the analysis of the microorganism or component thereof.
  • the analysis may include identification of the microorganism.
  • the analysis may be an in vitro or in vivo analysis, as hereinabove described.
  • the disease or condition is an infection with a Gram positive bacteria.
  • the pathogenic Gram positive pathogenic bacteria is selected from the group consisting of: Bacillus; Clostridium; Corynebacterium; Enterococcus; Listeria; Staphylococcus; and Streptococcus.
  • the Gram positive bacteria is a cocci e.g. Staphylococcus; or Streptococcus.
  • the Staphylococcus is S. aureus.
  • the S. aureus may be, without limitation thereto, a glycopeptide-sensitive strain (such as ATCC 25923); an MRSA stain (such as ATCC 43300); a glycopeptide-intermediate (GISA) strain (such as NRS 17); or a vancomycin-resistant (VRSA) strain (such as NARSA VRS4).
  • the Staphylococcus is S. epidermis.
  • the S. epidermis may be, without limitation thereto, a glycopeptide-sensitive strain (such as ATCC 12228); or glycopeptide-intermediate (GISE) strain (such as NARSA NRS 60).
  • the Streptococcus is S. pneumoniae.
  • the S. pneumoniae may be, without limitation thereto, a glycopeptide sensitive strain (such as ATCC 33400); or a glycopeptide resistant strain (such as ATCC 700677).
  • the Enterococcus is E. faecium.
  • the E. faecium may be, without limitation thereto, a vancomycin A (Van A) resistant strain (such as ATCC 51559) or a vancomycin B (Van B) resistant strain (such as ATCC 51299).
  • the Enterococcus is E. faecalis.
  • the E. faecalis may be, without limitation, a vancomycin sensitive strain (such as ATCC29212) or a vancomycin resistant strain.
  • the disease or condition may be selected from the group consisting of a bacterial infection of: the respiratory system (e.g. pneumonia); the digestive tract (e.g. gastroenteritis); the sinus (e.g. sinusitis); the ears (e.g. otitis media); the nervous system (e.g. meningitis); the skin (e.g. cellulitis); or the endocrine system (e.g. bacterial pancreatitis).
  • the respiratory system e.g. pneumonia
  • the digestive tract e.g. gastroenteritis
  • the sinus e.g. sinusitis
  • the ears e.g. otitis media
  • the nervous system e.g. meningitis
  • the skin e.g. cellulitis
  • the endocrine system e.g. bacterial pancreatitis
  • the disease or condition is bacteraemia. In another particularly preferred embodiment the disease or condition is bacterial sepsis. In one preferred embodiment, the disease or condition is bacterial sepsis caused by S. aureus.
  • the subject is a human or an animal subject as hereinabove described.
  • the subject is a human.
  • diagnosing and/or monitoring a condition may involve visualizing infections in vivo within any suitable location within the animal or human body. It will be appreciated that a disease or condition may be monitored by assessing replication or reproduction of a microorganism within the body.
  • diagnosing and/or monitoring a condition according to this aspect may involve identifying the location or source of an infection within the human or animal body by visualizing a microorganism according to the method of this aspect.
  • in vivo visualization may be used to identify a microorganism to identify contamination occurring during a surgical procedure.
  • in vivo visualization may be used to identify a potentially contaminated surgical implant that has been placed within the body.
  • Embodiments of the invention may have particular advantages with respect to time to analyse a microorganism or component thereof in a sample and/or to diagnosis a disease or condition based on this identification.
  • analysis of a microorganism or component thereof as per the directly preceding aspect of the invention can be performed in less than 10 hours; less than 9 hours; less than 8 hours; less than 7 hours; less than 6 hours; less than 5 hours; less than 4 hours; less than 3 hours; less than 2 hours; or less than 1 hours.
  • diagnosis of a disease or condition according to the method of this aspect can be performed in in less than 10 hours; less than 9 hours; less than 8 hours; less than 7 hours; less than 6 hours; less than 5 hours; less than 4 hours; less than 3 hours; less than 2 hours; or less than 1 hours.
  • a related aspect of the invention provides a method of treating a disease or condition, the method including the steps of diagnosing a disease or condition as hereinabove described, and treating the disease or condition based on the diagnosis. It will be appreciated that the particular form of treatment that will be suitable according to the method of this aspect will be related to the particular disease that is diagnosed. Often, in preferred embodiments wherein the disease is a Gram positive bacterial infection, the treatment will comprise administration of an antibiotic.
  • the antibiotic is selected from the group consisting of cloxacillin; dicloxacillin; methlocillin; nafcillin; oxacillin; cefazolin; cefoxitin; cefuroxime; cefepime; cefoperazone; cefotaxime; ceftazidime; ceftizoxime; ceftriaxone; trimethoprim; sulfamethoxazole; amoxicillin; clavulanate; penicillin; penicillin G; streptomycin; amoxicillin; clindamycin; doxycycline; etronidazole; rifampin; and vancomycin, or combinations thereof.
  • the antibiotic is vancomycin.
  • the antibiotic is selected from the group consisting of trimethoprim; sulfamethoxazole; clindamycin; vancomycin; doxycycline; minocycline; linezolid; and rifampin, or combinations thereof.
  • the treatment may additionally or alternatively involve surgical intervention.
  • diagnosis and initial treatment according to the method of this aspect can be performed in less than 10 hours; less than 9 hours; less than 8 hours; less than 7 hours; less than 6 hours; less than 5 hours; less than 4 hours; less than 3 hours; less than 2 hours; or less than 1 hour.
  • the invention also provides compounds comprising an optionally derivatized glycopeptide antibiotic bound to a first linker.
  • Such compounds may be referred to herein as 'glycopeptide antibiotic adducts'.
  • Such compounds of the invention will be suitable for connection to a visualization component, to form constructs as hereinabove described.
  • the glycopeptide antibiotic of such is selected from the group consisting of vancomycin; teicoplanin; oritavancin; telavancin; chloroeremomycin; and balhimycin.
  • the glycopeptide antibiotic is vancomycin.
  • the first linker of the second broad form comprises a PEG group, preferably at least PEG3.
  • the first linker comprises a linear carbon chain greater than four carbons, preferably wherein the linear carbon chain C4-C12.
  • the linear carbon chain is C8.
  • the linear carbon chain is CI 1.
  • the first linker comprises an amine-derived moiety.
  • the amine-derived group is at an end of the first linker.
  • the amine-derived moiety is an amide bond connecting the first linker to the glycopeptide antibiotic.
  • the first linker comprises an azide moiety.
  • the azide moiety is at an end of the linker opposite the glycopeptide antibiotic.
  • the invention provides a method of increasing or enhancing the activity or efficacy of a glycopeptide antibiotic, the method including the step of connecting a first linker to the glycopeptide antibiotic.
  • the invention provides a method of increasing or enhancing the activity or efficacy of a glycopeptide antibiotic, the method including the step of connecting the glycopeptide antibiotic to a visualization component using a first linker.
  • glycopeptide antibiotic is hereinabove described in relation to constructs of the invention.
  • glycopeptide antibiotics connected to particular linkers demonstrate increased activity towards at least certain microorganisms, as compared to the corresponding 'unconnected' or 'free' glycopeptide antibiotic.
  • constructs of the invention comprising particular linkers and a visualization component demonstrate increased activity towards at least certain microorganisms, as compared to the corresponding unconnected or free glycopeptide antibiotic of the construct.
  • the first linker comprises a linear carbon chain of at least four carbons.
  • the first linker comprises C8.
  • the activity or efficacy of the glycopeptide antibiotic is towards a
  • Gram positive bacteria Preferably the bacteria is a pathogenic Gram positive bacteria.
  • Particular preferred Gram positive bacteria according to this aspect are as described above in relation to methods of diagnosing a disease or condition according to the invention.
  • the Gram positive bacteria is a vancomycin resistant bacteria.
  • the vancomycin resistant bacteria may be a Van A or Van B resistant bacteria, although without limitation thereto.
  • the vancomycin resistant bacteria is selected from the group consisting of S. aureus; E. faecium; and E. faecalis.
  • the microorganism shows at least partial resistance to the free or unconnected glycopeptide antibiotic.
  • the increase or enhancement of the activity or efficacy or the antibiotic is a decrease in the Minimum Inhibitory Concentration (MIC) of the antibiotic for a particular microorganism, such as a Gram positive microorganism as described above.
  • MIC Minimum Inhibitory Concentration
  • the decrease in MIC is a fold decrease of between about 1.5 and about 5 to the free or unconnected antibiotic, including a fold decrease of at least: 2; 2.5; 3; 3.5; 4; or 4.5.
  • Another aspect of the invention provides a method of inhibiting, controlling, or killing a microorganism, the method including the steps of contacting a microorganism with (i) a construct of the invention; or (ii) a glycopeptide antibiotic adduct of the invention, to thereby inhibit, control, or kill the microorganism.
  • the microorganism is a Gram positive bacteria.
  • the Gram positive bacteria is a pathogenic Gram positive bacteria.
  • Particularly preferred Gram positive bacteria according to this aspect are as described above in relation to methods of diagnosing a disease or condition according to the invention.
  • compositions and Methods for Diagnosis, Treatment or Prevention of Disease are provided.
  • the invention provides a composition for diagnosing, treating or preventing a disease, disorder, or condition in a subject, the composition comprising (i) a construct of the invention; and/or (ii) a glycopeptide antibiotic adduct of the invention, as hereinabove described.
  • composition may suitably contain one or more pharmaceutically- acceptable carriers, diluents or excipients.
  • pharmaceutically -acceptable carrier, diluent or excipien is meant a solid or liquid filler, diluent or encapsulating substance that may be safely used in systemic administration.
  • a variety of carriers well known in the art, may be used.
  • These carriers may be selected from a group including sugars, starches, cellulose and its derivatives, malt, gelatine, talc, calcium sulphate, vegetable oils, synthetic oils, polyols, alginic acid, phosphate buffered solutions, emulsifiers, isotonic saline and salts such as mineral acid salts including hydrochlorides, bromides and sulphates, organic acids such as acetates, propionates and malonates and pyrogen-free water.
  • a useful reference describing pharmaceutically acceptable carriers, diluents and excipients is Remington's Pharmaceutical Sciences (Mack Publishing Co. N.J. USA, 1991) which is incorporated herein by reference.
  • compositions according to this aspect include tablets, dispersions, suspensions, injections, solutions, oils, syrups, troches, capsules, suppositories, aerosols, transdermal patches and the like. These dosage forms may also include injecting or implanting controlled releasing devices designed specifically for this purpose or other forms of implants modified to act additionally in this fashion. Controlled release of the therapeutic agent may be effected by coating the same, for example, with hydrophobic polymers including acrylic resins, waxes, higher aliphatic alcohols, polylactic and polyglycolic acids and certain cellulose derivatives such as hydroxypropylmethyl cellulose. In addition, the controlled release may be effected by using other polymer matrices, liposomes and/or microspheres.
  • compositions of the present invention suitable for enteral, oral or parenteral administration may be presented as discrete units such as capsules, sachets or tablets each containing a pre-determined amount of one or more therapeutic agents of the invention, as a powder or granules or as a solution or a suspension in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion or a water-in-oil liquid emulsion.
  • Such compositions may be prepared by any of the methods of pharmacy but all methods include the step of bringing into association one or more agents as described above with the carrier which constitutes one or more necessary ingredients.
  • the compositions are prepared by uniformly and intimately admixing the agents of the invention with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired presentation.
  • Any safe route of administration may be employed for providing a patient with constructs, adducts, or compositions of the invention.
  • enteral, oral, rectal, parenteral, sublingual, buccal, intravenous, intra-articular, intra-muscular, intra-dermal, subcutaneous, inhalational, intraocular, intraperitoneal, intracerebroventricular, transdermal and the like may be employed.
  • the construct, adduct or composition may be administered in a manner compatible with the dosage formulation, and in such amount as is pharmaceutically- effective.
  • the dose administered to a patient should be sufficient to effect a beneficial response in a patient over an appropriate period of time.
  • the quantity of agent(s) to be administered may depend on the subject to be treated inclusive of the age, sex, weight and general health condition thereof, factors that will depend on the judgement of the practitioner.
  • treatment methods and pharmaceutical compositions may be applicable to prophylactic or therapeutic treatment animals, inclusive of humans and non-human mammals such as livestock (e.g. horses, cattle and sheep), companion animals (e.g. dogs and cats), laboratory animals (e.g. mice, rats and guinea pigs) and performance animals (e.g. racehorses, greyhounds and camels), although without limitation thereto.
  • livestock e.g. horses, cattle and sheep
  • companion animals e.g. dogs and cats
  • laboratory animals e.g. mice, rats and guinea pigs
  • performance animals e.g. racehorses, greyhounds and camels
  • the disease or condition according to these embodiments is caused by a Gram positive bacteria.
  • Particular preferred Gram positive bacteria according to this aspect are as described above in relation to methods of diagnosing a disease or condition according to the invention.
  • the disease or condition may be selected from the group consisting of a bacterial infection of: the respiratory system (e.g. pneumonia); the digestive tract (e.g. gastroenteritis); the urinary tract (e.g. a urinary tract infection); the sinus (e.g. sinusitis); the ears (e.g. otitis media); the nervous system (e.g. meningitis); the skin (e.g. cellulitis); or the endocrine system (e.g. bacterial pancreatitis).
  • the respiratory system e.g. pneumonia
  • the digestive tract e.g. gastroenteritis
  • the urinary tract e.g. a urinary tract infection
  • the sinus e.g. sinusitis
  • the ears e.g. otitis media
  • the nervous system e.g. meningitis
  • the skin e.g. cellulitis
  • the endocrine system e.g. bacterial pancreatitis
  • the disease or condition is bacteraemia. In another particularly preferred embodiment the disease or condition is bacterial sepsis. In one preferred embodiment, the disease or condition is bacterial sepsis caused by a S. aureus.
  • This Example describes particular preferred constructs of the invention, comprising a glycopeptide antibiotic in the form of vancomycin; a first linker; a second linker; and a fluorophore.
  • the Example also describes particular preferred glycopeptide antibiotic adducts, comprising vancomycin and a first linker. As described, the glycopeptide antibiotic adducts were used for production of the constructs.
  • Commercially available cartridges were used for MPLC chromatography (Reveleris C18 Reversed-Phase 12 g cartridge and 40 g cartridge), while HPLC purifications used an Agilent Eclipse XDB-Phenyl column 30 x 100 mm, 5 ⁇ particle size.
  • 1 H (600 MHz) and 13 C (150 MHz) NMR spectra were obtained using a Bruker Avance-600 spectrometer equipped with a TXI cryoprobe.
  • the glycopeptide antibiotic vancomycin can be modified at a number of regions that do not substantially interfere with binding of vancomycin. These sites have accessible functional groups including the C-terminal carboxy group, primary and secondary amine groups, and hydroxyl and phenolic groups, all of which have been used to generate vancomycin derivatives. Binding of vancomycin depends mainly on the heptapeptide backbone. For this example, vancomycin was modified at the C-terminal carboxy group, as modification of this site should not interfere with binding, nor with vancomycin dimerization, an additional component of vancomycin's mode of action.
  • a Cu-catalyzed azide-alkyne cycloaddition (CuAAC) reaction strategy was used production for production of glycopeptide adducts.
  • the CuAAC reaction is compatible with the multiple unprotected functional groups presented in antibiotics, especially the amine, hydroxyl, and amide groups on vancomycin. Glycopeptide adducts containing vancomycin and different linkers were synthesized.
  • FIG. 2 illustrates the synthesis of the vancomycin adducts. Vancomycin adducts (2 and 3, respectively) were synthesized via amide coupling between an azido-alkyl-amine (N 3 -C8-NH 2 ) or azido-PEG3 -amine (N 3 -PEG3-NH 2) and the carboxy group of vancomycin.
  • Vancomycin (1) was reacted with azide-linkers in the presence of benzotriazol-l-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP) and ⁇ , ⁇ -diisopropylethylamine (DIPEA) in DMF, providing azide- derivatised vancomycin adducts.
  • PyBOP benzotriazol-l-yl-oxytripyrrolidinophosphonium hexafluorophosphate
  • DIPEA ⁇ , ⁇ -diisopropylethylamine
  • Vancomycin adduct 2 is hereinafter referred to as N 3 -C8-Van, or alternatively Van-8C-N 3 .
  • Vancomycin adduct 3 is hereinafter referred to as N 3 -PEG3-Van, or alternatively Van-PEG3-N .
  • - Characteristics of N 3 -C8-Van (2) obtained were as follows
  • (+)-ESI-HRMS calc for C7 4 H 93 Cl 2 Ni 3 02 3 [M+2H] 2+ : 800.7942, found 800.7907.
  • (+)-ESI-HRMS calc for C7 4 H 93 Cl 2 Ni 3 026 [M+2H] 2+ : 824.7866, found 824.7861.
  • Fluorescent vancomycin probes were prepared by a single step synthesis using vancomycin N 3 -C8-Van and N 3 -PEG3-Van, set forth in FIG. 2.
  • fluorophores 7- nitrobenzofurazan (NBD) and 7-(dimethylamino)-coumarin-4-acetic acid (DMACA) were utilised as they have low molecular weight and/or no electronic charges, and it was considered likely that these would cause minimal disturbance to antimicrobial activity. These fluorophores have previously been successfully used to prepare other fluorescent antibiotics [l]-[2].
  • Fluorophores NBD and DMACA were selected to prepare two colours (green and blue) of fluorescent vancomycin probes.
  • NBD-alkyne (labelled ⁇ ') and DMACA-alkyne (labelled ⁇ ⁇ ) precursors were produced.
  • a schematic of the production of 10 and 11 is provided in FIG. 2A. Details of the synthesis are as follows.
  • NBD-alkyne (10) To produce NBD-alkyne (10), to a solution of 4-chloro-7- nitrobenzo[c][l,2,5]oxadiazole (300 mg, 1.5 mmol) in THF (10 mL) was added a solution of propargyl amine (110 ⁇ , 1.65 mmol), Cs 2 C0 3 (480 mg, 1.5 mmol). The reaction mixture was stirred at 50 °C for 4 h. After completion of the reaction, the reaction mixture was diluted with EtOAc (50 mL), washed with H 2 0 (30 mL), brine (30 niL). The organic phase was separated, dried (MgS0 4 ), and evaporated to give the residue. The residue was purified by Si column chromatography (petroleum ether/EtOAc, 7:3) to give 10 (240 mg, 75%).
  • DMACA-alkyne (11) a solution of the 2-(7-(dimethylamino)-2- oxo-2H-chromen-4-yl)acetic acid (0.3 g, 1.21 mmol) in DMF (5 mL) was added HATU in DMF (5 mL) followed by DIPEA (386 ⁇ ), and propargylamine (71 ⁇ , 1.1 mmol). The solution was stirred at RT overnight. The reaction was evaporated under reduced pressure to remove DMF. The residue was diluted with water and extracted with ethyl acetate, dried over MgS0 4 , and concentrated under reduced pressure. The crude compound was recrystallized in CH 2 CI 2 . The solid was filtrated and washed with CH 2 C1 2 to give DMACA-alkyne (11) (0.149 g, 48%) as a green solid.
  • constructs (labelled ⁇ 2', ' 13' . and ' 14') were synthesised then using NBD-alkyne (10) and DMACA-alkyne (11), via CuAAC reactions.
  • acetic acid as a proton source accelerated the conversion of C-Cu bond-containing intermediate (Cu (I) acetylide and 5-cuprated 1,2,3-triazole) in CuAAC reaction, leading to reduction of byproduct since byproduct is increased by prolonging the reaction time.
  • the reaction mixture was stirred in a microwave reactor at 100 °C for 15 min.
  • the reaction mixture was concentrated under reduced pressure to yield the crude product.
  • the crude compounds were pre-purified by MPLC over C18 silica gel (Grace Reveleris, A: H 2 0 (0.1% TFA), B: ACN (0.1% TFA), 0 ⁇ 100% B over 8 min).
  • N 3 -C8-Van (150 mg, 9.37 x 10 "5 mol) was reacted with NBD-alkyne (10) to give visualization construct 12 in the form of an orange powder (34 mg, 20%).
  • Construct 12 is herein referred to as Vanco-8C-Tz-NBD, or alternatively NBD-Tz- C8-Van.
  • Construct 13 is herein referred to as Vanco-8c-Tz-DMACA, or alternatively DMACA-Tz-C8-Van.
  • N 3 -PEG3-Van (150 mg, 9.09 x 10 "5 mol) was reacted with NBD-alkyne (10) to give visualization construct 14 in the form of an orange powder (34 mg, 18%).
  • Visualization construct 14 is herein referred to as Vanco-PEG3-Tz-NBD, or alternatively NBD-Tz-PEG3-Van.
  • (+)-ESI-HRMS calc for C 83 HiooCl 2 Ni 7 0 2 6 [M+3H] 3+ : 606.8801, found 606.8801.
  • the synthesized constructs and glycopeptide adducts were tested for antimicrobial activity against a panel of Gram-positive bacteria including ATCC reference strains and clinical isolates of Staphylococcus aureus (S. aureus), Streptococcus pneumoniae (S. pneumoniae), Enterococcus faecalis (E. faecalis), and Enterococcus faecium (E. faecium) (FIG. 11).
  • Vancomycin and a vancomycin adduct comprising a first linker containing a 3 carbon linear chain (Van-3C-N 3 ) were included as controlled.
  • bacteria were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA), Merck Sharp & Dohme (Kenilworth, NJ), the Coli Genetic Stock Center (CGSC, Yale University), and independent academic clinical isolate collections. Bacteria were cultured in Muller Hinton broth (MHB) (Bacto Laboratories, Cat. no. 211443) at 37 °C overnight. A sample of each culture was then diluted 50-fold in MHB and incubated at 37 °C for 1.5-3 h. The constructs and compounds were serially diluted two-fold across the wells, with concentrations ranging from 0.06 ⁇ g/mL to 128 ⁇ g/mL, plated in duplicate.
  • ATCC American Type Culture Collection
  • VA Manassas, VA, USA
  • Merck Sharp & Dohme Kenilworth, NJ
  • CGSC Coli Genetic Stock Center
  • CGSC Coli Genetic Stock Center
  • MIC Minimum Inhibitory Concentration
  • NBD-Tz-C8-Van (12) and DMACA-Tz-C8-Van (13) showed generally increased activity against this Gram-positive panel compared to vancomycin, while NBD-Tz-PEG3-Van maintained antimicrobial potency (FIG. 11).
  • glycopeptide adduct N 3 -C8-Van exhibited generally increased activity against the Gram positive panel (FIG. 11), including increased activity against VRSA and VRE.
  • constructs of the invention comprising fluorescent probes with small molecular size and/or neutral charge coupled to a glycopeptide antibiotic (vancomycin) may be particularly suitable for binding to Gram positive bacteria.
  • constructs of the invention comprising such fluorescent probes linked to a glycopeptide antibiotic (vancomycin) by particular linkers (e.g. linkers comprising C8) may result in constructs with increased activity towards Gram positive bacteria, as compared to the corresponding free glycopeptide antibiotic.
  • SR-SIM Super-resolution structured illumination microscopy
  • constructs showed strong fluorescence at the dividing septum compared to the lateral wall, confirming binding of the construct via the glycopeptide antibiotic to the nascent peptidoglycan Lipid II.
  • Constructs comprising linkers comprising PEG3 (i.e. NBD-Tz-PEG3-Van; 14) and C8 (i.e. NBD-Tz-C8-Van; 12) did not show substantial differences in selective binding. Notably however, construct 12 was approximately 4-fold more active than construct 14 against the Gram-positive panel.
  • Vancomycin probes have been used to study mechanism of bacterial cell division such as co-labelling with PG synthesis proteins [5], fluorescence microscopy of vancomycin-labelled bacteria showed the dynamics of peptidoglycan assembly in ovococci [6] and the PG biosynthesis in B. subtilis [10].
  • septum is the structure that forms in the middle of the mother cell by invagination of the cell membrane and ingrowth of the cell wall, which splits the mother cell into two identical daughter cells [8]
  • septum is the structure that forms in the middle of the mother cell by invagination of the cell membrane and ingrowth of the cell wall, which splits the mother cell into two identical daughter cells [8]
  • the vancomycin probes are able to spatially discriminate the peptidoglycan layer from the bacterial cell membrane as shown by cross-section measurements in FIG. 14A, which clearly show the peak of probe intensity outside of the peak intensity of a membrane- selective probe.
  • vancomycin probes are potentially useful for assessing the target sites of other antibiotic probes such as TMP and linezolid probes.
  • Co-staining of DMACA-Tz-C8-Van (13) (blue) with TMP and linezolid NBD probes (green) showed that the linezolid probe and TMP probe penetrated within the bacterial cell membrane into the cytosol.
  • Both TMP and linezolid generally showed co- localisation with FM4-64FX (membrane dye) indicating that the membrane dye was quickly endocytosed in S. aureus (FIG. 14B-C).
  • HXPI The structure of HXPI is given in FIG. 15(A).
  • DOTA-alkyne To produce chelating compound 2,2',2"-(10-(2-oxo-2-(prop-2-yn-l- ylamino)ethyl)- 1 ,4,7, 10-tetraazacyclododecane- 1 ,4,7-triyl)triacetic acid (DOTA- alkyne), The NHS ester of DOTA (104.7 mg, 0.137 mmol) was dissolved in 3 mL DMF under argon, then propargylamine (17 ⁇ , 0.27 mmol) was added. The yellow solution was agitated at room temperature overnight, then concentrated under reduced pressure to give a yellow oil.
  • DOTA-alkyne The structure of DOTA-alkyne is given in FIG. 15(C).
  • N 3 -C8-Van adduct was produced as follows. Vancomycin-HCl (511 mg, 0.344 mmol) was dissolved in 25 mL DMF, and PyBOP (199 mg, 0.383 mmol) was added, followed by DiPEA (0.44 mL, 2.53 mmol). 8- Azido-octylamine (142 mg, 0.686 mmol) was dissolved in 1 mL DMF, then added to the vancomycin solution. The reaction was stirred at room temperature for 16 h, then quenched with MeOH and concentrated to roughly half the original volume under reduced pressure. The concentrated reaction was injected directly onto a MPLC reverse phase column (0 - 100% ACN (0.1% TFA) in water (0.1% TFA)) to give vanco-C 8 -N 3 as a white solid.
  • the blue mixture was cooled and injected directly onto a MPLC reverse phase column (0 - 100% ACN (0.1% FA) in water (0.1% FA)) to give semi-pure vanco-Cg-PXPI as a blue solid.
  • PXPI-Tz-PEG3-Van was produced similarly as described for PXPI-Tz-C8-Van, with the exception that N 3 -PEG3-Van was used in place of N 3 -C8-Van.
  • DOTA-C8-Van was produced by dissolving N 3 -C8- Van (10.13 mg, 0.00632 mmol) in 6 mL DMF, and DOTA-alkyne (13.83 mg, 0.0314 mmol) was added and stirred at 50°C for 30 min. Cul (25.26 mg, 0.133 mml), then DiPEA (130 ⁇ L, 0.75 mmol), then AcOH (130 ⁇ L, 2.2 mmol) were added and the reaction stirred for 10 mins. The reaction was cooled and injected directly onto a MPLC reverse phase column (0 - 100% ACN (0.1% FA) in water (0.1% FA)) and all peaks after the initial 0% flush were combined and lyophilised.
  • the resulting solid was dissolved in 1: 1 v/v ACN/H 2 0, and Na 2 S-9H 2 0 was added. The suspension was vortexed and centrifuged to precipitate copper sulfide species. This process was repeated until no more precipitation was observed.
  • DOTA-Tz-PEG3-Van was produced similarly as described for DOTA-Tz-C8-Van, with the exception that N 3 -PEG3-Van was used in place of N 3 -C8-Van.
  • TMP trimethoprim
  • erythromycin and citropin 1.1 and the vancomycin HCl and NBD-Tz- PEG3-Van controls
  • 32 ⁇ g/ml for all other antibiotics were 64 ⁇ g/ml for erythromycin and citropin 1.1 (and the vancomycin HCl and NBD-Tz- PEG3-Van controls) and 32 ⁇ g/ml for all other antibiotics.
  • the compound was serially diluted two-fold across the wells of 96-well micro- titre plates (corning; Cat. No 3370, non-treated polystyrene plate) and NBS 96-well micro-titre plates (corning; Cat. No 3461, non-binding surface), with concentrations ranging from 0.06 ⁇ g/mL to 128 ⁇ g/mL, plated in duplicate.
  • the resultant mid-log phase culture was diluted to 1 x 10 6 CFU/mL, then 50 ⁇ , was added to each well of the compound-containing 96-well plates giving a cell density of 5 x lO 5 CFU/mL, and a final compound concentration range of 0.03 ⁇ g/mL to 64 ⁇ g/mL for citropin 1.1 and of 0.015 ⁇ g/mL to 32 ⁇ g/mL for other antibiotics.
  • the plates were covered and incubated at 37 °C for 22 h. MICs were determined visually, being defined as the lowest concentration showing no visible growth. MICs were determined visually at 22 h incubation and the MIC was defined as the lowest concentration with which no growth was visible after incubation.
  • NBD-Tz-PEG3-Van at 32 ⁇ g/mL in HBSS: 500 ⁇ , for one sample 2.
  • Each antibiotic compound at 0.125, 1.25, 6.25, 12.5, 125 g/mL in HBSS: 1 mL for one sample.
  • E. coli were cultured in LB at 37°C overnight. A sample of each culture was then diluted 50-fold in LB and incubated at 37°C for 1.5-2 h. The resultant mid log phase cultures were harvested at 4000 rpm for 15 min, washed once with HBSS (4000 rpm, for 15 min), and resuspended in HBSS to an OD600 of 1.
  • Results of the fluorescence intensity assessment using flow cytometry are set out in Figure 18.
  • a summary of assay results for the eight antibiotic compounds PMXB (polymyxin B) sulphate, Colistin sulphate, OCT C4 TFA (octapeptin C4 trifluoroacetate salt), Tachyplesin-1, Arenicin-3, Gentamicin, TMP, Citropin 1.1, and Erythromycin is set forth in Table 2.

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Abstract

L'invention concerne une construction de visualisation comprenant : (i) un antibiotique glycopeptidique éventuellement dérivé; (ii) un composant de visualisation; et (iii) un premier lieur liant (i) et (ii). Le composant de visualisation peut être un composant fluorescent, un point quantique, un composant de visualisation IRM, un composant de visualisation PET ou SPECT, un composant de visualisation MPI, ou un composant de visualisation radiographique. L'antibiotique glycopeptidique peut être la vancomycine, la téicoplanine, l'oritavancine, la télavancine, la chloroérémomycine ou la balhimycine. Le premier lieur peut comprendre une fraction polyéthylène glycol (PEG), ou une chaîne carbonée linéaire de plus de quatre carbones. L'invention concerne également des procédés associés de production et d'utilisation de composés, d'adduits et de constructions connexes, tels que des procédés de visualisation de micro-organismes.
PCT/AU2017/051365 2016-12-09 2017-12-11 Constructions de visualisation WO2018102890A1 (fr)

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CN110643034A (zh) * 2018-06-26 2020-01-03 湖南华腾医药有限公司 一种多臂型peg化奥利万星衍生物及其制备
WO2021069685A1 (fr) * 2019-10-10 2021-04-15 Universite De Strasbourg Analogue fluorescent de la vancomycine
WO2021086982A3 (fr) * 2019-10-28 2021-08-05 Beckman Coulter, Inc. Composés d'identification de classes microbiennes et leurs utilisations

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CN110643034A (zh) * 2018-06-26 2020-01-03 湖南华腾医药有限公司 一种多臂型peg化奥利万星衍生物及其制备
CN110643034B (zh) * 2018-06-26 2022-05-06 湖南华腾医药有限公司 一种多臂型peg化奥利万星衍生物及其制备
WO2021069685A1 (fr) * 2019-10-10 2021-04-15 Universite De Strasbourg Analogue fluorescent de la vancomycine
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WO2021086982A3 (fr) * 2019-10-28 2021-08-05 Beckman Coulter, Inc. Composés d'identification de classes microbiennes et leurs utilisations

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