WO2019088917A1

WO2019088917A1 - Imidazolium-quaternary ammonium copolymers as novel antibacterial and antifungal materials

Info

Publication number: WO2019088917A1
Application number: PCT/SG2018/050495
Authority: WO
Inventors: Yugen Zhang; Yuan Yuan
Original assignee: Agency for Science Technology and Research Singapore
Current assignee: Agency for Science Technology and Research Singapore
Priority date: 2017-11-01
Filing date: 2018-09-28
Publication date: 2019-05-09
Anticipated expiration: 2020-05-01
Also published as: CN111344333A; SG11202002936YA

Abstract

The present invention relates to quaternary ammonium-imidazolium copolymers having the following Formula (I): where the various groups are as defined in the specification. The present invention also relates to the polymer having the following Formula (II): where the various groups are as defined in the specification. The present invention also relates to the methods for their preparation, antimicrobial composition, antimicrobial gel containing these polymers of Formulae (I) and (II), and uses of these polymers in the treatment of a microbial infection or disease.

Description

Title of Invention: Imidazolium-Quaternary

Ammonium Copolymers as Novel Antibacterial and Antifungal Materials Cross- Reference to Related Application

This application claims priority to Singapore application number 10201709027U filed on 1 November 2017, the disclosure of which is hereby incorporated by reference.

Technical Field

The present invention generally relates to a polymer of Formula (I) or a polymer of Formula (II). The present invention also relates to a composition, a pharmaceutical composition, methods of producing the polymer of Formula (I) or the polymer of Formula (II), the uses and methods of uses thereof.

Background Art

Infectious diseases and the increasing threat of worldwide pandemics have underscored the importance of antibiotics and hygiene. Although many classes of antibiotics and antimicrobial agents have been developed and used, the emergence of antimicrobial resistance in patients and the environment urges the development of novel antimicrobial materials that circumvent or reduce selection for resistant microbial strains.

Among the various types of antimicrobial agents, the cationic compounds, such as quaternary ammonium compounds (QACs), are the most valuable antiseptics and disinfectants currently. QACs exert antibacterial activity against both Gram-positive and Gram-negative bacteria, as well as against some pathogenic species of fungi and protozoa. However, over the last few years, the development and applications of QACs have been reduced due to the emergence of antimicrobial resistance and potential toxicity toward mammalian cells and the ecosystem.

Recently, imidazolium salts have emerged as new alternatives for antimicrobial applications. Di-imidazolium salts showed good antimicrobial activity and low toxicity to mammalian cells. The selectivity can be tuned by modifying the structure of imidazolium with different functional groups or changing the anions. A series of main-chain imidazolium oligomers and polymers have been developed, which demonstrated high efficacy and high selectivity against a broad range of bacteria and fungi. These polymers and oligomers were designed to capture the essential features of antimicrobial peptides, such as amphiphilic structure of cationic hydrophilic groups and hydrophobic moieties.

In fact, polycationic materials provide a valid approach to address both of the resistance and toxicity problems. Synthetic polymers that target the membranes of many pathogenic species are reported to have low susceptibility for developing resistance, unlike small molecular antibiotics and conventional low-molecular- weight QACs. In topical applications, cationic polymers have limited residual toxicity since they are more difficult to permeate through skin. Compared to low-molecular-weight QACs, cationic polymers have higher positive charge density which promotes initial adsorption onto the negatively charged bacterial surfaces and disruption of cellular membranes, resulting in significantly enhanced antibacterial activity. However, a drawback for some of the cationic polymers is that they can lead to hemolysis, which is one of the more harmful side effects of many cationic polymers.

Accordingly, there is a need for a new series of polymers that have antimicrobial properties and which address or alleviate one or more disadvantages mentioned above.

Summary

According to a first aspect, there is provided a polymer having the following Formula (I):

Formula (I)

wherein

L¹, L² and L³ are independently selected from the group consisting of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted alkenylalkyl, optionally substituted alkylalkenyl, optionally substituted alkylalkenylalkyl, optionally substituted arylalkyl, optionally substituted arylalkenyl, optionally substituted arylalkynyl, optionally substituted alkylaryl, optionally substituted alkenylaryl, and optionally substituted alkynyl aryl;

X is independently selected from a halogen;

n, m and p are independentiy an integer of at least 1 ;

q is 0 or an integer of at least 1 ;

A has the following structure:

wherein R¹, R², R³ and R⁴ are independently selected from an optionally substituted alkyl, or any two of R¹, R², R³ and R⁴ may be taken together to form at least one bridging group, and

x and y are independently an integer of at least 1 , or a salt or hydrate thereof. Advantageously, the ammonium-imidazolium copolymers as defined herein may display antimicrobial activity against a broad range of microbe or microorganism. More advantageously, the ammonium-imidazolium copolymer may also display antifungal activity a broad range of fungal species. Most of the ammonium-imidazolium copolymers may show much higher activity against fungi compared to single component imidazolium or ammonium polymers. More advantageously, the copolymer may be biocompatible and degradable with non-resistance property. These copolymers may be non-hemolytic.

According to another aspect, there is provided a composition comprising the polymer as defined herein or a salt or hydrate thereof, in association with a carrier.

Advantageously, the ammonium-imidazolium copolymers may not suffer from antimicrobial resistance. More advantageously, the ammonium-imidazolium copolymers may be used against microbe or microorganism that has developed a resistance to conventional antimicrobial drugs.

According to another aspect, there is provided use of the polymer as defined herein or the composition as defined herein as a non-therapeutic agent for killing or inhibiting the growth of a microorganism.

According to another aspect, there is provided a pharmaceutical composition comprising the polymer as defined herein or a pharmaceutically acceptable salt or hydrate thereof, in association with a pharmaceutically acceptable carrier.

According to another aspect, there is provided a method for killing or inhibiting the growth of a microorganism, the method comprising administering to a subject the polymer as defined herein or the pharmaceutical composition as defined herein.

According to another aspect, there is provided a polymer as defined herein or a pharmaceutical composition as defined herein, for killing or inhibiting the growth of a microorganism.

According to another aspect, there is provided use of the polymer as defined herein or the pharmaceutical composition as defined herein, in the manufacture of a medicament for killing or inhibiting the growth of a microorganism.

According to another aspect, there is provided a method for treating a microbial infection, the method comprising administering to a subject the polymer as defined herein or the pharmaceutical composition as defined herein.

According to another aspect, there is provided a polymer as defined herein or a pharmaceutical composition as defined herein for use as an antibiotic.

According to another aspect, there is provided use of the polymer as defined herein or the pharmaceutical composition as defined herein, in the manufacture of a medicament for treating a microbial infection.

According to another aspect, there is provided a method for preparing the polymer as defined herein, comprising the steps of:

contacting a diamine having the following structure:

R², R³ and R⁴ are independently selected from an optionally substituted alkyl, or any two of R¹, R², R³ and R⁴ is taken together to form at least one bridging group; and x is an integer of at least 1 ;

with a di-imidazole having the following structure:

ortho -phenylene group, pcra-phenylene group, meia-phenylene group and ethenylene;

and a dihalide having the following structure:

, wherein L⁵ is selected from the group consisting of orf/io-phenylene group,

/?ara-phenylene group, mefa-phenylene group and ethenylene; and X is a halide;

under reaction conditions.

Advantageously, these copolymers may be easy and straightforward to synthesize with relatively low cost.

Definitions

The following words and terms used herein shall have the meaning indicated:

The term "polymer" as used herein refers to a large molecule, or macromolecule, composed of a number of repeating units, up to 30 in total of the same repeating units, whereby the repeating unit may be ammonium, imidazolium, any linkers (L¹, L² and L³) of Formula (I), any linkers (L⁴ and L⁵) of Formula (II) or any combinations thereof.

"Alkyl" as a group or part of a group refers to a straight or branched aliphatic hydrocarbon group to be interpreted broadly, having from 1 to 16 carbon atoms, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 carbon atoms, preferably a C₁-C16 alkyl, C₁-C₁₂ alkyl, more preferably a C₁-C₁₀ alkyl, most preferably Ci-Ce alkyl unless otherwise noted. Examples of suitable straight and branched alkyl substituents include but is not limited to, methyl, ethyl, 1 -propyl, isopropyl, 1-butyl, 2-butyl, isobutyl, tert-butyl, amyl, 1 ,2-dimethylpropyl, 1, 1- dimethylpropyl, pentyl, isopentyl, hexyl, 4-methylpentyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3- dimethylbutyl, 5-methylheptyl, 1-methylheptyl, octyl, nonyl, decyl, undecyl, 2,2,3- trimethyl-undecyl, dodecyl, 2,2-dimethyl-dodecyl, tridecyl, 2-methyl-tridecyl, 2-methyl- tridecyl, tetradecyl, 2-methyl-tetradecyl, pentadecyl, 2-methyl-pentadecyl, hexadecyl, 2- methyl-hexadecyl and the like. The alkyl may be optionally substituted with one or more groups as defined under the term "optionally substituted" below.

"Alkenyl" as a group or part of a group denotes an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched preferably having 2 to 16 carbon atoms, e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or 16 carbon atoms, preferably a Ci-C₁₆alkenyl, Q-C^alkenyl, more preferably a C Ci₀alkenyl, most preferably Cj-Cealkenyl in the normal chain. The group may contain a plurality of double bonds in the normal chain and the orientation about each is independently E or Z. Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl and nonenyl. The group may be a terminal group or a bridging group. The bridging group may be ethenylene or vinylene. The alkenyl may be optionally substituted with one or more groups as defined under the term "optionally substituted" below.

"Alkynyl" as a group or part of a group means an aliphatic hydrocarbon group containing a carbon-carbon triple bond and which may be straight or branched preferably having from 2 to 16 carbon atoms, e.g. 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or 16 carbon atoms, preferably a Q-C16 alkynyl, Q-C₁₂ alkynyl, more preferably a Cj-Cio alkynyl, most preferably C₁-C6 alkynyl in the normal chain. Exemplary structures include, but are not limited to, ethynyl and propynyl. The group may be a terminal group or a bridging group. The alkynyl may be optionally substituted with one or more groups as defined under the term "optionally substituted" below.

"Aryl" as a group or part of a group to be interpreted broadly denotes (i) an optionally substituted monocyclic, or fused polycyclic, aromatic carbocycle (ring structure having ring atoms that are all carbon) preferably having from 5 to 12 atoms per ring, e.g. 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms, wherein the optionally substitution can be di-substitution, or tri- substitution. Examples of aryl groups include phenyl, naphthyl, and the like; (ii) an optionally substituted partially saturated bicyclic aromatic carbocyclic moiety in which a phenyl and a C5-7 cycloalkyl or C_{5 7} cycloalkenyl group are fused together to form a cyclic structure, such as tetrahydronaphthyl, indenyl or indanyl. The group may be a terminal group or a bridging group. Typically an aryl group is a C₆-Ci₈ aryl group. The aryl may be optionally substituted with one or more groups as defined under the term "optionally substituted" below. Preferably the groups may include orfAo-phenylene group, para-phenylene group and mefa-phenylene group where it is used interchangeably with o-phenylene group, /?-phenylene group and m-phenylene group.

"Alkylaryl" refers to an alkyl-aryl group in which alkyl and aryl moieties are as defined herein. Alternatively, "arylalkyl" refers to an aryl-alkyl group in this sequence, in which aryl and alkyl moieties are as defined herein. Preferred alkylaryl groups are Ci-C _t-alkylaryl having 6 or 10 carbon atoms in the aryl. Preferred arylalkyl groups are aryl-Ci-C₄-alkyl having 6 or 10 carbon atoms in the aryl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the aryl group. The alkyl moiety of the alkylaryl or arylalkyl may also be the terminating molecule.

"Alkenylalkyl" refers to an alkenyl-alkyl group in which alkenyl and alkyl moieties are as defined herein. Alternatively, "alkylalkenyl" refers to an alkyl -alkenyl group in this sequence, in which alkyl and alkenyl moieties are as defined herein. Preferred alkenylalkyl groups are C₂-Ce~ alkenylalkyl having 1 to 10 carbon atoms in the alkyl. Preferred alkylalkenyl groups are alkyl-C₂-C₆-alkenyl having 1 to 10 carbon atoms in the alkyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the aryl group. "Alkylalkenylalkyl" refers to alkyl-alkenyl-alkyl group in which alkenyl and alkyl moieties are as defined herein. Preferred alkylalkenylalkyl groups are alkylC₂-C₃-alkenylalkyl having 1 to 10 carbon atoms in the alkyl. Preferred alkylalkenylalkyl groups are alkyl-C₂- Ce- alkenyl- alkyl having 1 to 10 carbon atoms in the alkyl. The group may be a terminal group or a bridging group.

"Alkenylaryl" refers to an alkenyl-aryl group in which alkenyl and aryl moieties are as defined herein. Alternatively, "arylalkenyl" refers to an aryl-alkenyl group in this sequence, in which aryl and alkenyl moieties are as defined herein. Preferred alkenylaryl groups are C₂-C6-alkenylaryl having 6 or 10 carbon atoms in the aryl. Preferred arylalkenyl groups are aryl-C2-C6-alkenyl having 6 or 10 carbon atoms in the aryl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the aryl group. The alkenyl moiety of the alkenylaryl or arylalkenyl may also be the terminating molecule.

"Alkynylaryl" refers to an alkynyl-aryl group in which alkynyl and aryl moieties are as defined herein. Alternatively, "arylalkynyl" refers to an aryl-alkynyl group in this sequence, in which aryl and alkynyl moieties are as defined herein. Preferred alkynylaryl groups are C₂-C₆- alkynylaryl having 6 or 10 carbon atoms in the aryl. Preferred arylalkynyl groups are aryl-C₂- C₆-alkynyl having 6 or 10 carbon atoms in the aryl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the aryl group. The alkynyl moiety of the alkynylaryl or arylalkynyl may also be the terminating molecule.

A "bond" is a linkage between atoms in a compound or molecule. The bond may be a single bond, a double bond, or a triple bond.

"Archaea" refers to a domain of single-celled microorganisms that do not have any cell nucleus or any other organelles inside their cells.

"Bacterium" refers to a member of a large group of unicellular microorganisms which have cell walls but lack a nuclear membrane or membrane-bound organelles and an organized nucleus, including some which can cause disease. Bacteria (plural) are categorized as gram-positive or gram-negative when a cell wall is present. While many bacteria are aerobic requiring the presence of oxygen to survive, others are anaerobic and are able to survive only in the absence of oxygen. Bacterium is any of a domain of chiefly round, spiral, or rod-shaped single-celled prokaryotic microorganisms that typically live in soil, water, organic matter, or the bodies of plants and animals, that make their own food especially from sunlight.

"Bridging group" refers to a group having from 2 to 50 atoms not counting hydrogen atoms, preferably 2 to 40 atoms, 2 to 30 atoms, 2 to 20 atoms, e.g. more preferably 2 to 20 carbon atoms, more preferably 2 to 16 carbon atoms, e.g. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15 or 16 carbon atoms, a Q-Cie alkyl, Ci-C₁₂ alkyl, more preferably a Q-Cio alkyl, most preferably C -Ce alkyl in the normal chain. The alkyl group may be divalent alkyl, alkenyl, alkynyl, aryl group but not limited to this. Exemplary alkyl groups include, but are not limited to, ethenylene (-CH₂CH₂-) group, ortho-phenylene group, para-phenylene group or meta-phenylene group. The bridging group may be optionally substituted with one or more groups as defined under the term "optionally substituted" below. "Fungus" refers to any member of the group of eukaryotic organisms that includes microorganisms such as yeasts and molds.

"Halide" or "halogen" represents chlorine, fluorine, bromine or iodine.

"Microorganism" or "microbe" being used interchangeably refers to an organism that is microscopic (too small to be visible to the naked eye). Microorganisms are often described as single-celled, or unicellular organisms.

"Polydispersity index or value" or "dispersity index (D)" refers is a measure of the distribution of molecular mass in a given polymer sample. D (PDI) of a polymer is calculated: PDI = M M_m where M_w is the weight average molecular weight and M_n is the number average molecular weight. M_n is more sensitive to molecules of low molecular mass, while M_w is more sensitive to molecules of high molecular mass. The dispersity indicates the distribution of individual molecular masses in a batch of polymers. D has a value equal to or greater than 1, but as the polymer chains approach uniform chain length, D approaches unity (1).

"Protist" refers to any eukaryotic organism or a diverse collection of organisms that is not an animal, plant or fungus. The protists do not form a natural group, or clade, since they exclude certain eukaryotes; but, like algae or invertebrates, they are often grouped together for convenience. While exceptions exist, they are primarily microscopic and unicellular, or made up of a single cell. The cells of protists are highly organized with a nucleus and specialized cellular machinery called organelles.

The term "xylylene" as used herein, is used interchangeably with "xylene".

The term "optionally substituted" as used herein means the group to which this term refers may be unsubstituted, or may be substituted with one or more groups independently selected from alkyl, alkenyl, alkynyl, thioalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkylalkenyl, heterocycloalkyl, cycloalkylheteroalkyl, cycloalkyloxy, cycloalkenyloxy, cycloamino, halo, carboxyl, haloalkyl, haloalkynyl, alkynyloxy, heteroalkyl, heteroalkyloxy, hydroxyl, hydroxyalkyl, alkoxy, thioalkoxy, alkenyloxy, haloalkoxy, haloalkenyl, haloalkynyl, haloalkenyloxy, nitro, amino, nitroalkyl, nitroalkenyl, nitroalkynyl, nitroheterocyclyl, alkylamino, dialkylamino, alkenylamine, aminoalkyl, alkynylamino, acyl, alkyloxy, alkyloxyalkyl, alkyloxyaryl, alkyloxycarbonyl, alkyloxycycloalkyl, alkyloxyheteroaryl, alkyloxyheterocycloalkyl, alkenoyl, alkynoyl, acylamino, diacylamino, acyloxy, alkylsulfonyloxy, heterocyclic, heterocycloalkenyl, heterocycloalkyl, heterocycloalkylalkyl, heterocycloalkylalkenyl, heterocycloalkylheteroalkyl, heterocycloalkyloxy, heterocycloalkenyloxy, heterocycloxy, heterocycloamino, haloheterocycloalkyl, alkylsulfinyl, alkylsulfonyl, alkylsulfenyl, alkylcarbonyloxy, alkylthio, acylthio, aminosulfonyl, phosphorus -containing groups such as phosphono and phosphinyl, sulfinyl, sulfinylamino, sulfonyl, sulfonylamino, aryl, heteroaryl, heteroarylalkyl, heteroarylalkenyl, heteroarylheteroalkyl, heteroarylamino, heteroaryloxy, arylalkenyl, alkenylaryl, arylalkyl, alkylaryl, alkylheteroaryl, aryloxy, arylsulfonyl, cyano, cyanate, isocyanate, -C(0)NH(alkyl), and -C(0)N(alkyl)₂. Where the term "substituted" is used, the group to which this term refers to may be substituted with one or more of the same groups mentioned above. The word "substantially" does not exclude "completely" e.g. a composition which is "substantially free" from Y may be completely free from Y. Where necessary, the word "substantially" may be omitted from the definition of the invention.

Unless specified otherwise, the terms "comprising" and "comprise", and grammatical variants thereof, are intended to represent "open" or "inclusive" language such that they include recited elements but also permit inclusion of additional, unrecited elements.

As used herein, the term "about", in the context of concentrations of components of the formulations, typically means +/- 5% of the stated value, more typically +/- 4% of the stated value, more typically +/- 3% of the stated value, more typically, +/- 2% of the stated value, even more typically +/- 1% of the stated value, and even more typically +/- 0.5% of the stated value.

Throughout this disclosure, certain embodiments may be disclosed in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosed ranges. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

Certain embodiments may also be described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the disclosure. This includes the generic description of the embodiments with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

Detailed Disclosure of Optional Embodiments

Exemplary, non-limiting embodiments of a polymer having the following formula (I) will now be disclosed.

The polymer may have the following formula (I):

Formula (I)

wherein

X is independently selected from a halogen;

n, m and p are independently an integer of at least 1 ;

q is 0 or an integer of at least 1;

A has the following structure:

wherein R¹, R², R³ and R⁴ are independently selected from an optionally substituted alkyl, or any two of R¹, R², R³ and R⁴ may be taken together to form at least one bridging group, and x and y are independently an integer of at least 1 , or

a salt or hydrate thereof.

In the polymer having the formula (I), L¹, L² and L³ may be independently selected from the group consisting of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted alkenylalkyl, optionally substituted alkylalkenyl, optionally substituted alkylalkenylalkyl, optionally substituted arylalkyi, optionally substituted arylalkenyl, optionally substituted arylalkynyl, optionally substituted alkylaryl, optionally substituted alkenylaryl, and optionally substituted alkynylaryl. The carbons atoms of alkyl group may be in the range of 1 to 20 carbon atoms, 1 to 16 carbon atoms, 1 to 12 carbon atoms, 1 to 10 carbon atoms or preferably 1 to 6 carbon atoms. The carbons atoms of alkenyl group or alkynyl group may be in the range of 2 to 20 carbon atoms, 2 to 16 carbon atoms, 2 to 12 carbon atoms, 2 to 10 carbon atoms or preferably 2 to 6 carbon atoms. The carbons atoms of aryl group may be in the range of 5 to 18 carbon atoms, 5 to 12 carbon atoms, 5 to 10 carbon atoms, 6 to 18 carbon atoms or preferably 6 to 12 carbon atoms. The aryl group may be selected from the group consisting of o-xylylene, m-xylylene and /7-xylylene.

The alkenyl-alkyl group may be C₂_₆alkenyl-Ci_₆alkyl group. The alkyl-alkenyl group may be C_halky 1-C₂_₆alkenyl group. The alkyl-alkenyl-alkyl group may be Ci_₆alkyl-C₂-6^alk^enyl-C₁_ ₆alkyl group. The aryl-alkyl group may be C6-i₂aryl-Ci_6alkyl group. The alkyl-aryl group may be C^alkyl-Ce-^aryl group. The aryl-alkenyl may be C6-i₂aryl-C_{2 6}alkenyl group. The alkenyl-aryl may be C₂_6alkenyl-C6 i₂aryl group. The aryl-alkynyl may be C6 ₁₂aryl-C₂_ ealkynyl group. The alkynyl-aryl may be C2-6alkynyl-C6 i₂aryl group. The alkyl- aryl- alkyl group may be CYealkyl-phenyl-C^alkyl. The C_{1 6}alkyl group may be methyl, ethyl, propyl, butyl, pentyl or hexyl. The phenyl of the Ci_6alkyl-phenyl-Ci_6alkyl group may be selected from the group consisting of o-xylylene, m-xylylene, 7-xylylene.

In the polymer having the formula (I), X may be a halogen from Group VII of the periodic table. X may be a halide or halogen selected from fluoride, chloride, bromide and iodide. In the polymer having the formula (I), n, m and p may be independently an integer of at least 1, or an integer between 1 to 5,000, between 1 to 1,000, between 1 to 500, between 1 to 100, preferably between 1 to 50, that is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50, more preferably between 1 to 10 or most preferably between 1 to 5. p may preferably be an integer of 1.

In the polymer having the formula (I), q may be 0 or may be an integer of at least 1, or an integer from 1 to 10,000, from 1 to 5,000, from 1 to 1,000, from 1 to 500, from 1 to 100, from 1 to 50, preferably from 1 to 25, that is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25. q may preferably be an integer of 1.

In the polymer having the formula (I), A may be of the following structure:

wherein R¹, R², R³ and R⁴ may be independently selected from an optionally substituted alkyl, or any two of R¹, R², R³ and R⁴ may be taken together to form at least one bridging group, and x and y may independently be an integer of at least 1. The optionally substituted alkyl may be optionally substituted Cusalkyl group, preferably optionally substituted Ci ^alkyl group or more preferably, optionally substituted Q^alkyl group. The any two of R¹, R², R³ and R⁴ may be taken together to form at least one bridging group. R¹ and R² may be taken together to form at least one bridging group. R³ and R⁴ may be taken together to form at least one bridging group. The bridging group may be C₂ to C₁₀ bridging group, preferably C₂ to C5 bridging group or more preferably an ethenylene group. The bridging group may be optionally substituted.

The x and y may independently be an integer of at least 1 or at least between 1 to 10, between 1 to 5, from 1, 2, 3, 4 or 5, or more preferably 1.

In the polymer having the formula (I), A may be selected from the group consisting of the following structures:

In the polymer having the formula (I), the molar ratio between A and L¹ may be in the range of 1 :5 and 5:1, in the range of 1:4 and 5:1, in the range of 1:3 and 5: 1, in the range of 1:2 and 5: 1, in the range of 1 : 1 and 5:1, in the range of 2: 1 and 5: 1, in the range of 3:1 and 5: 1, in the range of 4:1 and 5: 1, in the range of 1:5 and 4:1, in the range of 1 :5 and 3:1, in the range of 1 :5 and 2: 1 or in the range of 1:5 and 1:1, in the range of 1 :5 and 1 :2, in the range of 1 :5 and 1:3 or in the range of 1 :5 and 1:4.

In the polymer having the formula (I), the molar ratio between (L¹ + L²) and L³ is in the range of about 1:5 to about 5:1, in the range of 1:4 and 5:1, in the range of 1 :3 and 5: 1, in the range of 1:2 and 5: 1, in the range of 1: 1 and 5:1, in the range of 2: 1 and 5:1, in the range of 3: 1 and 5:1, in the range of 4:1 and 5: 1, in the range of 1:5 and 4:1, in the range of 1:5 and 3:1, in the range of 1:5 and 2:1 or in the range of 1 :5 and 1 :1, in the range of 1 :5 and 1 :2, in the range of 1 :5 and 1 :3 or in the range of 1 :5 and 1:4.

In the polymer having the formula (I), the polymer may have a molecular weight in the range of about 1,000 to about 20,000, about 1,000 to about 15,000, about 1,000 to about 10,000, about 1,000 to about 9,000, about 1,000 to about 8,000, about 1 ,000 to about 7,000, about 1,000 to about 6,000, about 1,000 to about 5,000, about 1,000 to about 4,000, about 1,000 to about 3,000, about 1,000 to about 2,000, about 2,000 to about 10,000, about 3,000 to about 10,000, about 4,000 to about 10,000, about 5,000 to about 10,000, about 6,000 to about 10,000, about 7,000 to about 10,000, about 8,000 to about 10,000, about 9,000 to about 10,000, about 10,000 to about 20,000, about 10,000 to about 15,000 or about 15,000 to about 20,000. The polymer having the formula (I) may have a high molecular weight when the polymer is synthesized in dimethylformamide (DMF).

In the polymer having the formula (I), the polymer may have a polydispersity value in the range of about 1.2 to about 3.2, about 1.4 to about 3.2, about 1.6 to about 3.2, about 1.8 to about 3.2, about 2.0 to about 3.2, about 2.2 to about 3.2, about 2.4 to about 3.2, about 2.6 to about 3.2, about 2.8 to about 3.2, about 3.0 to about 3.2, about 1.2 to about 3.0, about 1.2 to about 2.8, about 1.2 to about 2.6, about 1.2 to about 2.4, about 1.2 to about 2.2, about 1.2 to about 2.0, about 1.2 to about 1.8, about 1.2 to about 1.6 or about 1.2 to about 1.4.

Advantageously, the polymer having the formula (I) may be characterized by relatively low degree of polymerization (M_w<10,000) and high polydispersity values (1.3 < D < 3.1).

The polymer may have the following formula (II):

Formula (II)

wherein

L⁴ and L⁵ are independently selected from optionally substituted alkenyl or optionally substituted aryl. The optionally substituted alkenyl may be optionally substituted C₂ isalkenyl group, optionally substituted C₂ i₂alkenyl group or preferably optionally substituted C₂ ₆alkenyl group. The optionally substituted alkenyl may be ethenylene. The optionally substituted aryl may be optionally substituted Ce-isaryl group, optionally substituted Ce ^aryl group or preferably optionally substituted Cearyl group. The optionally substituted aryl may be selected from the group consisting of ort/zo-phenylene (o-phenylene) group, /¾¾ra-phenylene (/7-phenylene) group and meto-phenylene (m-phenylene) group.

In the polymer having the formula (II), X may be a halogen from Group VII of the periodic table. X may be a halide or halogen selected from fluoride, chloride, bromide and iodide.

In the polymer having the formula (II):

be selected from the group consisting of the following structures:

In the polymer having the formula (II), p and q may be independently an integer between 1 to 20, between 1 to 10, between 1 to 5, from 1, 2, 3, 4 or 5, or more prelerably 1.

The polymer having the formula (I) or formula (II) may be selected from the group consisting of:

The distribution of molecular weights of the polymer having the formula (I) may be affected by the solubility of the polymer in solvents. The polymer having the formula (I) may have higher solubility in DMF than in THF due to the higher polarity of DMF.

Advantageously, the ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) may display synergistic effect when ammonium and imidazolium are combined together instead of the individual ammonium polymer or imidazolium polymer. Preferably, DABCO and imidazolium may be combined together as the ammonium-imidazolium copolymers.

Advantageously, the ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) may display antimicrobial activity against a broad range of microbe or microorganism. More advantageously, the ammonium-imidazolium copolymer may also display antifungal activity a broad range of fungal species. Most of the ammonium-imidazolium copolymers may show much higher activity against fungi compared to single component imidazolium or ammonium polymers.

The minimum inhibitory concentrations (MICs) of ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) against bacteria may be in the range of about 1 to about 200 μg/mL, about 1 to about 180 μg/mL or about 1 to about 160 μg/mL.

Preferably, the minimum inhibitory concentrations (MICs) of ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) against bacteria (E. coli) may be in the range of about 10 to about 150 μg/mL, about 10 to about 125 μg/mL, about 10 to about 100 μg/mL, about 10 to about 80 μg/mL, about 10 to about 60 μg/mL, about 10 to about 40 μg mL, preferably about 10 to about 30 μg/mL or more preferably about 10 to about 16 μg/mL. The ammonium-imidazolium copolymer may preferably be TMED-imidazolium copolymer with -phenylene linker or DMP -imidazolium copolymer with /?ara-phenylene linker. More preferably, the ammonium-imidazolium copolymer may be DABCO-imidazolium copolymer with irans-butene linker.

Preferably, the minimum inhibitory concentrations (MICs) of ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) against bacteria (5. aureus) may be in the range of about 5 to about 80 μg/mL, about 5 to about 60 μg/mL, about 5 to about 50 μg/mL, about 5 to about 40 μg/mL, about 5 to about 30 μg/mL, about 5 to about 20 μg/mL, preferably about 5 to about 16 μg/mL or more preferably about 8 μg mL. The ammonium-imidazolium copolymer may preferably be TMED-imidazolium copolymer with ara-phenylene linker or DMP-imidazolium copolymer with £>ara-phenylene linker. More preferably, the ammonium- imidazolium copolymer may be DABCO-imidazolium copolymer with trans-b tene linker.

Preferably, the minimum inhibitory concentrations (MICs) of ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) against bacteria (P. aeruginosa) may be in the range of about 10 to about 80 μg/mL, about 10 to about 60 μg/mL, about 10 to about 40 μg/mL, about 10 to about 30 pg/mL, about 10 to about 25 μg/mL, about 10 to about 20 μg mL, preferably about 31 μg/mL, or more preferably 16 μg/mL. The ammonium-imidazolium copolymer may preferably be DMP-imidazolium copolymer with /jara-phenylene linker. More preferably, the ammonium-imidazolium copolymer may be DABCO-imidazolium copolymer with trans-butene linker.

The minimum inhibitory concentrations (MICs) of ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) against fungi species may be in the range of about 1 to about 200 μg/mL, about 1 to about 180 μg/mL, about 1 to about 160 μg/mL or about 1 to about 140 μg/mL.

Preferably, the minimum inhibitory concentrations (MICs) of ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) against fungi species (C. albicans) may be in the range of about 1 to about 130 μg/mL, about 1 to about 115 μg/mL, about 1 to about 100 μg/mL, about 1 to about 70 μg/mL, about 1 to about 40 μg/mL, about 1 to about 20 μg/mL₎ about 1 to about 10 μg/mL, preferably about 1 to about 8 μg/mL or more preferably about 2 μg/mL. The ammonium-imidazolium copolymer may preferably be DMP based polymer with frares-butene linker. More preferably, the ammonium-imidazolium copolymer may be DABCO- imidazolium copolymer with fraras-butene linker.

Preferably, the minimum inhibitory concentrations (MICs) of ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) against fungi species (F. solani) may be in the range of about 20 to about 130 μg mL, about 20 to about 100 μg/mL, about 20 to about 80 μg/mL, about 20 to about 60 μg/mL, about 20 to about 50 μg mL, about 20 to about 40 μg/mL, about 20 to about 35 μg/mL or more preferably about 31 μg/mL. The ammonium- imidazolium copolymer may preferably be TMED-imidazolium copolymer with para- phenylene linker. More preferably, the ammonium-imidazolium copolymer may be DABCO- imidazolium copolymer with iraras-butene linker.

Advantageously, the ammonium-imidazolium copolymers may not suffer from antimicrobial resistance. More advantageously, the ammonium-imidazolium copolymers may be used against microbe or microorganism that has developed a resistance to conventional antimicrobial drugs. More advantageously, the DABCO-imidazolium copolymer with iraras-butene linker may have better minimum inhibitory concentrations (MICs) than conventional antimicrobial drugs. The minimum inhibitory concentrations (MICs) may be in the range of about 1 to about 10 μg/mL, preferably about 1 to about 8 pg/mL or more preferably about 2 pg/mL.

The minimum fungicidal concentration (MFC) of DABCO-imidazolium copolymer with trans- butene linker may be in the range of about 50 to about 90 pg/mL, about 50 to about 80 μg mL, about 50 to about 70 μg/mL, preferably about 55 to about 65 pg/mL or more preferably 62 pg/mL. The ammonium-imidazolium copolymers as defined by Formula (I) or Formula (II) may have better stability than the conventional antimicrobial drugs. Advantageously, the copolymer may be biocompatible and non-hemolytic.

More advantageously, these copolymers may be easy and straightforward to synthesize and relatively low cost. Exemplary, non-limiting embodiments of a composition comprising the polymer as defined herein will now be disclosed.

The composition may comprise of the polymer as defined herein, or a salt or hydrate thereof, in association with a carrier.

Exemplary, non-limiting embodiments of the use of the polymer or the composition as defined herein will now be disclosed.

The use of the polymer as defined herein or the composition as defined herein, may be as a non- therapeutic agent for killing or inhibiting the growth of a microorganism. Alternatively, the polymer may be used as a therapeutic agent for killing or inhibiting the growth of a microorganism.

Exemplary, non-limiting embodiments of a pharmaceutical composition will now be disclosed.

The pharmaceutical composition may comprise of the polymer as defined herein, or a pharmaceutically acceptable salt or hydrate thereof, in association with a pharmaceutically acceptable carrier or a pharmaceutically acceptable excipient.

The polymer may be administered alone or in the form of a pharmaceutical composition in combination with a pharmaceutically acceptable carrier, diluent or excipient.

The amount of the polymer in the composition may be such that it is effective to measurably treat the microbial infection or disease. The amount of polymer in the composition may be such that it is effective to measurably treat the disease, disorder or condition associated with microbial infection. The amount of polymer in the composition may be such that it is effective to measurably treat the disease, disorder or condition associated with microbial infection that is from any of the microorganisms. The composition may be formulated for administration to a subject in need of such composition. The composition may be formulated for administration to a patient in need of such composition. In using the polymers, they may be administered in any form or mode which may make the polymer bioavailable. One skilled in the art of preparing formulations can readily select the proper form and mode of administration depending upon the particular characteristics of the polymer selected, the condition to be treated, the stage of the condition to be treated and other relevant circumstances. The pharmaceutically acceptable carrier or pharmaceutically acceptable excipient may be a nontoxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the polymer with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the composition may include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene - polyoxypropylene-block polymers, polyethylene glycol or wool fat.

Compositions or pharmaceutical compositions as defined above may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this disclosure may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed including synthetic mono- or di- glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

Pharmaceutically acceptable compositions as defined above may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

Pharmaceutical compositions for parenteral injection may comprise pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate. Proper fluidity may be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservative, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of micro-organisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminium monostearate and gelatin. If desired, and for more effective distribution, the polymers may be incorporated into slow release or targeted delivery systems such as polymer matrices, liposomes, and microspheres.

The injectable formulations may be sterilized, for example, by filtration through a bacterial- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use.

Alternatively, pharmaceutically acceptable compositions as defined above may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

Pharmaceutically acceptable compositions as defined above may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations may be readily prepared for each of these areas or organs. Topical application for the lower intestinal tract may be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutically acceptable compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of compounds as defined above may include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutically acceptable compositions may be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers may include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2 octyldodecanol, benzyl alcohol and water. For ophthalmic use, the pharmaceutically acceptable compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutically acceptable compositions may be formulated in an ointment such as petrolatum.

Pharmaceutically acceptable compositions as defined above may also be administered by nasal aerosol or inhalation. Such compositions may be prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.

Most preferably, pharmaceutically acceptable compositions as defined above may be formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions as defined above may be administered without food. In other embodiments, pharmaceutically acceptable compositions as defined above may be administered with food.

The amount of compound that may be combined with the carrier materials to produce a composition in a single dosage form may vary depending upon the host treated, the particular mode of administration. Preferably, the compositions should be formulated so that a dosage of between 0.01 - 100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a polymer of the present disclosure in the composition will also depend upon the particular compound in the composition.

Exemplary, non-limiting embodiments of a method for killing or inhibiting the growth of a microorganism will now be disclosed.

The method for killing or inhibiting the growth of a microorganism, the method may comprise of administering to a subject the polymer as defined herein, or the pharmaceutical composition as defined herein.

The subject may be a cell. The subject may be a human or animal body. The cell may be present in a cell culture in vitro. The cell may be from a cell line. The cell line may be an immortalized cell line, a genetically modified cell line or a primary cell line. The cell may be from a tissue of a subject. The cell may be in a subject.

The method of killing or inhibiting the growth of the microorganism may be a non-therapeutic method whereby the polymer may be formulated as a disinfectant, a sterilizing agent or a surface cleaning agent. The polymer may be used to create an antiseptic environment. Exemplary, non-limiting embodiments of a polymer or a pharmaceutical composition as defined herein will now be disclosed.

The polymer as defined herein or the pharmaceutical composition as defined herein may be for killing or inhibiting the growth of a microorganism. The polymer as defined herein or the pharmaceutical composition as defined herein may be used for killing or inhibiting the growth of a microorganism.

Exemplary, non-limiting embodiments of a use of the polymer or the pharmaceutical composition as defined herein will now be disclosed.

There is provided use of the polymer as defined herein or the pharmaceutical composition as defined herein may be in the manufacture of a medicament for killing or inhibiting the growth of a microorganism.

The microorganism may be a bacterium, archaea, fungus, protist, animal, plant, or any mixture thereof.

The microorganism may be a disease causing microorganism. The disease may be a microbial infection or microbial disease. The disease causing microorganism may be selected from the group consisting of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Fusarium solani and Candida albicans.

Exemplary, non-limiting embodiments of a method for treating a microbial infection will now be disclosed.

The method for treating a microbial infection may comprise the step of administering to a subject the polymer as defined above or the pharmaceutical composition as defined above. The polymer or pharmaceutical composition may treat the microbial infection or disease caused by a microorganism.

Exemplary, non-limiting embodiments of a polymer or a pharmaceutical composition as defined herein will now be disclosed.

The polymer as defined above or a pharmaceutical composition as defined above may be for use as an antibiotic. The antibiotic may be for treating a microbial infection or disease. The antibiotic may be used for treating a microbial infection or disease.

There is provided use of the polymer as defined above or the pharmaceutical composition as defined above may be in the manufacture of a medicament for treating a microbial infection.

The microbial infection or disease may be caused by a bacterium, archaea, fungus, protist, animal, plant, or any mixture thereof. The microbial infection or disease may also be caused by a microbe or microorganism.

The microbe or microorganism may be selected from the group consisting of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Fusarium solani and Candida albicans. The method for inhibiting or method for treating may be an in vitro method. The method of inhibiting or method of treating may also be an in vivo method.

Exemplary, non-limiting embodiments of a method for preparing a polymer having the following formula (II) will now be disclosed.

The method for preparing a polymer as defined herein may comprise the steps of:

contacting a diamine having the following structure:

, wherein R¹, R², R³ and R⁴ are independently selected from an optionally substited alkyl, or any two of R¹, R², R³ and R⁴ is taken together to form at least one bridging group; and x is an integer of at least 1 ;

with a di-imidazole having the following structure:

wherein L is selected from the group consisting of ortho phenylene group, para-phenylene group, meta-phenylene group and ethenylene;

and a dihalide having the following structure:

, wherein L is selected from the group consisting of orfAo-phenylene group, group, meia-phenylene group and ethenylene; and X is a halide; under reaction conditions.

The diamine having the may be selected from the group

diamine may not be limited to these examples.

The di-imidazole having the following

method may be selected from the group consisting

imidazole may not be limited to these examples.

iodide or fluoride.

The X of the dihalide may be a halide or halogen from Group VII of the periodic table. X may be a halide or halogen selected from fluoride, chloride, bromide and iodide.

The reaction conditions of the above method may comprise the temperature at which the reaction is stirred at, the solvent that the reaction is stirred in or the time period or duration that is required for the reaction to be completed.

The temperature of the reaction may be an elevated temperature. The elevated temperature may be in the range of about 60 °C to about 100 °C, about 65 °C to about 100 °C, about 70 °C to about 100 °C, about 75 °C to about 100 °C, about 80 °C to about 100 °C, about 85 °C to about 100 °C, about 90 °C to about 100 °C, about 95 °C to about 100 °C, about 60 °C to about 95 °C, about 60 °C to about 90 °C, about 60 °C to about 85 °C, about 60 °C to about 80 °C, about 60 °C to about 75 °C, about 60 °C to about 70 °C or about 60 °C to about 65 °C. The temperature may preferably be in the range of about 70 to 90 °C.

The time period or duration of the reaction may be in the range of about 18 hours to 60 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, about 44 hours, about 46 hours, about 48 hours, about 50 hours, about 52 hours, about 54 hours, about 56 hours, about 58 hours, about 60 hours, preferably about 18 hours, or more preferably about 24 hours.

The solvent of the reaction may be an organic solvent. The solvent may be a polar solvent or non-polar solvent. The solvent may preferably be a polar organic solvent. The polar organic solvent may be selected from the group consisting of tetrahydrofuran (THF), dimethylformamide (DMF), acetone, dimethyl sulfoxide (DMSO), acetonitrile, dicholormethane and ethyl acetate (EtOAc). The polar organic solvent may preferably be tetrahydrofuran (THF) dimethylformamide (DMF) or any mixture thereof.

The yields of the various copolymers from the above method or reaction may be in the range of about 45% to about 95%, about 49% to about 95%, about 55% to about 95%, about 60% to about 95%, about 65% to about 95%, about 70% to about 95%, about 75% to about 95%, about 80% to about 95%, about 85% to about 95%, about 90% to about 95%, about 45% to about 90%, about 45% to about 85%, about 45% to about 80%, about 45% to about 75%, about 45% to about 70%, about 45% to about 65%, about 45% to about 60%, about 45% to about 55% or about 45% to about 49%.

The diamine and di -imidazole may be contacted at a molar ratio in the range of about 1:5 to about 5: 1, in the range of 1 :4 and 5:1, in the range of 1 :3 and 5: 1, in the range of 1:2 and 5: 1, in the range of 1 : 1 and 5:1, in the range of 2:1 and 5: 1, in the range of 3:1 and 5: 1, in the range of 4: 1 and 5:1, in the range of 1:5 and 4:1, in the range of 1:5 and 3: 1, in the range of 1:5 and 2: 1 or in the range of 1:5 and 1 :1, in the range of 1:5 and 1 :2, in the range of 1 :5 and 1:3 or in the range of 1:5 and 1 :4. Preferably, the molar ratio may be 1 :3, 1 : 1 or 3: 1.

The di-imidazole and dihalide may be contacted at a molar ratio in the range of about 1:6 to about 5:6, in the range of 1 :5 and 5:6, in the range of 1 :4 and 5:6, in the range of 1:3 and 5:6, in the range of 1 :2 and 5:6, in the range of 1 :1 and 5:6, in the range of 2:1 and 5:6, in the range of 3: 1 and 5:6, in the range of 4: 1 and 5:6, in the range of 5: 1 and 5:6, in the range of 5:2 and 5:6, in the range of 5:3 and 5:6, in the range of 5:4 and 5:6, in the range of 5(1):5(1) and 5:6, in the range of 1 :5 and 4:6, in the range of 1 :5 and 3:6, in the range of 1:5 and 2:6 or in the range of 1:5 and 1 :6, in the range of 1 :5 and 5(1):5(1), in the range of 1:5 and 5:4, in the range of 1 :5 and 5:3, in the range of 1 :5 and 5:2 or in the range of 1:5 and 5:1.. Preferably, the molar ratio may be 1 :4, 1 :2 or 3:4.

The di-imidazole, diamine and dihalide may be contacted in any combinations thereof other than mentioned above, at a molar ratio as defined above.

Brief Description of Drawings

The accompanying drawings illustrate a disclosed embodiment and serves to explain the principles of the disclosed embodiment. It is to be understood, however, that the drawings are designed for purposes of illustration only, and not as a definition of the limits of the invention. Fig. 1

[Fig. 1] shows the antifungal activity of IBN-132-3 against C. albicans. Fig. 1A shows the killing efficacy of IBN-132-3 at low concentrations (from 0.5 MIC to 4 MIC, MIC = 2 μg/mL) against C. albicans after 48 hours of treatment. Fig. IB shows the colony forming unit of C. albicans after 24 hours or 48 hours of treatment with IBN-132-3 (T), Amphotericin B (A) and Fluconazole (F) at different concentrations

C. albicans grown in Yeast Mold broth were used as control. Circles indicate no colony observed. The data are expressed as mean ± S.D. of triplicates. *p<0.05; **p<0.01.

Fig. 2

[Fig. 2] shows the resistance acquisition in the presence of sub-MIC levels of copolymers and antibiotics against 5. aureus (Fig. 2A) and C. albicans (Fig. 2B).

Fig. 3

[Fig. 3] is a series of Scanning Electron Microscopy (SEM) images of C. albicans treated with copolymers (125 μg/mL) at room temperature for 24 hours. Fig. 3 A is the control, Fig. 3B is IBN-131-2, Fig. 3C is IBN-132-3, Fig. 3D is IBN-212. The cell wall was disrupted after exposure. C. albicans treated with YMB were used as controls. Scale bars represent 5 μηι.

Fig. 4

[Fig. 4] shows a series of antimicrobial activities of the copolymers against biofilms. Cell viability remained in C. albicans (Fig. 4A), E.coli (Fig. 4B), S. aureus (Fig. 4C) and P. aeruginosa (Fig. 4D) biofilms treated with different copolymers for 24 hours. The data are expressed as mean ± S.D. of quadruplicates. *statistical significance versus control, *p<0.05; **p<0.01.

Examples

Non-limiting examples of the invention will be further described in greater detail by reference to specific Examples, which should not be construed as in any way limiting the scope of the invention.

List of abbreviations used

ACN: acetonitrile

AcOEt: ethyl acetate

Ac OH: acetic acid

NH₄CI: Ammonium chloride

AUC: area under the curve

Brine: saturated aqueous solution of NaCl

bs: broad signal (broad peak) NMR

cat.: catalyst

Cs₂C0₃: Cesium carbonate

CH₂CI₂ or DCM: Methylene chloride or Dichloromethane

DAB CO : 1 ,4-diazabicyclo- [2.2.2] -octane

DBU: l,8-Diazabicycloundec-7-ene

DI: deionized water

DMF: A^A -dimethylformamide

DMSO: dimethylsulf oxide

DMSO-d₆: per-deuterated dimethylsulfoxide

DMP: 1,4-Dimethylpiperazine

Ether: diethylether

EtOH: ethanol

H: hour(s)

HPLC: high pressure liquid chromatography

IPA: Iso-propanol (2-propanol)

KOH: Potassium hydroxide

L: litre(s)

LC-MS: Liquid chromatography-mass spectrometry

Me: methyl

MeOH: methanol

m.p.: melting point

min: minute(s)

MS: mass spectrometry

Et₃N: triethylamine

Na₂C0₃: Sodium carbonate

NaHC0₃: Sodium bicarbonate

NaH: Sodium hydride NaOH: Sodium hydroxide

Na₂S0₄: Sodium sulphate

NMR: Nuclear Magnetic Resonance

PBS: Phosphate buffered saline

Rt: room temperature

TEA: triethylamine

THF: tetrahydrofuran

TLC: thin layer chromatography

TMED: tetramethylethane- 1 ,2-diamine

Materials and Methods

All anhydrous solvents were purchased from Sigma-Aldrich Corp. (St. Louis, Missouri, U.S.A.) and used without further purification. All other reagents were used as received, except where otherwise noted in the experimental text below.

Analytical thin layer chromatography (TLC) was performed using Merck 60 F-254 silica gel plates with visualization by ultraviolet light (254 nm) and/or heating the plate after staining with a solution of 20% KMn04 w/v in H₂0. Flash column chromatography was carried out on Kieselgel 60 (0.040-0.063 mm) supplied by Merck (Burlington, Massachusetts, U.S.A.) under positive pressure.

!H and ¹³C nuclear magnetic resonance (NMR) spectra were recorded on Bruker AV-400 (400 MHz) spectrometer. Chemical shifts (δ) are reported in parts per million (ppm) with the residual solvent peak of tetramethylsilane used as the internal standard at 0.00 ppm. *H NMR data are reported in the following order: chemical shift, multiplicity (br = broad, s = singlet, d = doublet, t = triplet, q = quartet and m = multiplet), integration and assignment. A Waters 2695 gel permeation chromatography (GPC) equipped with two ultrahydrogel columns: 300 mm x 7.8 mm in series and a Waters 2414 differential refractometer detector (Waters Corporation, Massachusetts, U.S.A.) were used to determine the molecular weights of the polymers and copolymers. Details of the mobile phase used are as follows: 54/23/23 (v/v/v%) 0.5 M sodium acetate aqueous solution/methanol/acetic acid, at a flow rate of 1.0 mL/min. Calibration was performed using a series of poly(ethylene glycol) standards of varying molecular weights (633- 20,600) (Polymer Standard Service Inc., Rhode Island, U.S.A.). Mw and D was subsequently calculated from the calibration curve.

Example 1

Structural Design

The copolymers were synthesized by introducing bis-quaternary ammonium components into poly-imidazolium chain to enhance the structural versatility and achieve optimal antibacterial/antifungal activity. Here, three a, co-tertiary diamines including 1 ,4-diazabicyclo- [2.2.2] -octane (DABCO), tetramethylethane-l,2-diamine (TMED) and 1,4-dimethylpiperazine (DMP) were chosen to form bis-quaternary ammonium. The diamines and di-imidazoles were randomly connected by benzylic or allylic dihalide compounds (either butene or xylylene linkers, Scheme 1). With this structural design, various ammonium-imidazolium random copolymers were synthesized. General structure:

( T) = ammonium = (X) x = CI or Br

(^ ) ( 2) = alkyl/aryl linkers

Specific structure:

Scheme 1. Structural design of random ammonium-imidazolium copolymers.

Synthesis of Di-Imidazoles

Scheme 2. Synthesis procedure of di -imidazoles

l,4-bis(N-imidazole-l-ylmethyl)benzene (la). NaOH (0.5 g, 12.5 mmol) was added to a dimethyl sulfoxide (DMSO) solution of imidazole (0.9 g, 13.0 mmol), and the resulting suspension was stirred at 90 °C for 2 hours, α,α '-dichloro-/>-xylylene (0.99 g, 5.7 mmol) was added to the residue. The resulting solution was stirred at 40 °C for 1 hour. The solvent was removed under vacuum. The product was extracted with dichloromethane (DCM), and la was obtained in quantitative yield after removing the solvent. *H NMR (CDC1₃): δ 7.55 (s, 2H), 7.13 (s, 4H), 7.10 (s, 2H), 6.89 (s, 2H), 5.12 (s, 4H). MS (GC-MS) m/z 238 (M⁺).

l,2-bis(N-imidazole-l-ylmethyl)benzene (lb), α,α'-dichloro-o-xylylene was used in the reaction instead of a,a'-dichloro- ?-xylylene to synthesize the desired named product (lb) in quantitative yield. Example 2

Synthesis of Polymers and Ammonium-Imidazolium Copolymers

General procedure for the synthesis of polymers and copolymers

The synthesis of ammonium-imidazolium copolymers was carried out by mixing di-imidazole, α,ω-diamine and dihalide linker in polar organic solvents, such as THF and DMF, at 70 to 90 °C for at least 24 hours. Modest to good yields ranging from 49% to 95% were obtained for various copolymers. Using DABCO as diamine, six copolymer samples (IBN-111, IBN-112, IBN-121, IBN-122, IBN-131 and IBN-132) with different linkers were synthesized (Scheme 3). In addition, the amount of DABCO to di-imidazole ratios in selected copolymers (IBN-131 and IBN-132) was adjusted to pursue higher antimicrobial activity.

An example of a specific procedure for the synthesis of copolymer IBN-132-3 (THF) is provided below:

DABCO (0.336 g, 3.0 mmol) and lb (0.238 g, 1.0 mmol) were dissolved in 5 mL of THF. A solution of trans-\ , dibromo-2-butene (0.856 g, 4.0 mmol) in DMF (5 mL) was then added dropwise at room temperature. After the addition was completed, the reaction mixture heated up to 70 °C and stirred for 24 hours. The co-polymer was obtained by centrifugation (5000 rpm, 5 minutes) and washed with acetone (3 x 15 mL) to afford a white powder in 85% yield.

l,4-diazabicyclo-[2.2.2]-octane (DABCO) solo polymers (IBN-110, IBN-120 and IBN-130) were also synthesized using a method similar to the above. The individual polymers (IBN-110, IBN-120 and IBN-130) were used as comparative polymers for testing against the ammonium- imidazolium copolymers.

The remaining polymers (IBN-110, IBN-120 and IBN-130) and copolymers (IBN-111, IBN- 112, IBN-121, IBN-122, IBN-131 and IBN-132) may be synthesized accordingly to the respective diamine, di-imidazole and aryl/alkyl linkers as indicated in the final compounds as well as the molar ratios as described in the scheme below (Scheme 3).

-112

R = DABCO; R = DABCO; LI, L2 = p-phenylene; R = DABCO; L1 , L2 = p-phenylene;

L1 = p-phenylene; X = Br L3 = p-phenylene; X = Br L3 = o-phenylene; X = Br

R:(L1+L2):L3 = 1:2:1 R:(L1+L2):L3 = 1:2:1

R = DABCO; R = DABCO; L1, L2 = o-phenyl R = DABCO; L1 , L2 = o-phenyl

L1 = o-phenylene; X = CI L3 = p-phenylene; X = CI L3 = o-phenylene; X = CI

R:(L1+L2):L3 = 1:2:1 R:(L1+L2):L3 = 1:2:1

-131 -132

R = DABCO; R = DABCO; L1 , L2 = frans-ethylene; R = DABCO; L1, L2 = frans-ethylene;

L1 = frans-ethylene; X = Br L3 = p-phenylene; X = Br L3 = o-phenylene; X = Br

-131-1: R:(L1 +L2):L3 = 1:4:3 -132-1: R:(L1+L2):L3 = 1:4:3 -131-2: R:(L1+L2):L3 = 1:2:1 -132-2: R:(L1+L2):L3 = 1:2:1 -131-3: R:(L1 +I_2):L3 = 3:4:1 -132-3: R:(L1+L2):L3 = 3:4:1

Scheme 3. Structures of DABCO based polymers and DABCO-imidazolium copolymers.

IBN-110: ¾ NMR (400MHz, DMSO-d₆): δ 7.56-7.80 (m, 4 nH, Fhfl), 4.50-5.08(m, 4 nH, DABCO-C/i₂-Ph), 2.94-4.19 (d, 12 nH, DABCO N⁺-CH₂-).

IBN-111: ¾ NMR (400MHz, DMSO-de): δ 9.30-9.73 (m, 2 nH, Im C2H), 6.90-7.94 (m, 16 nH, Im C4H, Im C5H, PhH), 5.18-5.62 (m, 8 nH, Ph-C¾- Im), 4.52-4.96 (m, 4 nH, Ph-C#₂- DABCO), 2.93-3.97 (m, 12 nH, DABCO N⁺-C¾-).

IBN-112: ¾ NMR (400MHz, DMSO-de): δ 9.19-9.68 (m, 2 nH, Im C2H), 6.91-7.94 (m, 16 nH, Im C4-H, Im C5H, PhH), 5.34-5.75 (m, 8 nH, Ph-Ci ₂-Im), 4.50-5.13 (m, 4 nH, Ph-C#₂- DABCO), 2.93-3.99 (m, 12 nH, DABCO N⁺-C¾-).

IBN-120: *H NMR (400MHz, DMSO-d₆): δ 7.40-8.10 (m, 4 nH, PhH), 4.67-5.69 (m, 4 nH, DABCO-C#₂-Ph), 3.09-4.11 (m, 12 nH, DABCO N⁺-CH₂-).

IBN-121: ¾ NMR (400MHz, DMSO-de): δ 9.56-9.86 (m, 2 nH, Im C2H), 6.99-8.04 (m, 16 nH, Im C4ff, Im C5H, PhH), 5.42-5.77 (m, 8 nH, Ph-C¾-Im), 4.92-5.28 (m, 4 nH, Ph-CH₂- DABCO), 2.98-4.06 (m, 12 nH, DABCO N⁺-C¾-).

IBN-122: ¾ NMR (400MHz, DMSO-de): δ 9.53-9.88 (m, 2 nH, Im C2H), 7.01-7.93 (m, 16 nH, Im-C4H, Im-C5ff, PhH), 4.62-5.88 (m, 12 nH, Ph-CH₂- Im, Ph-C/f₂-DABCO), 2.98-4.06 (m, 12 nH, DABCO-N⁺-C¾-).

IBN-130: ^JH NMR (400MHz, DMSO-d₆): δ 6.16-6.50 (m, 2 nH, -CH₂-C//=C#-CH₂-), 4.30- 4.64 (s, 4 nH, -CH₂-C//=C#-CH₂-), 3.00-4.11 (m, 12 nH, DABCO N⁺-CH₂-).

IBN-131-2: ¾ NMR (400MHz, DMSO-d₆): δ 9.61 -9.68 (m, 2 nH, Im C2H), 7.86-7.99 (m, 4 nH, Im C H, Im C5H), 7.56 (m, 4 nH, PhH), 6.00-6.44 (m, 4 nH, -CH=CH-), 5.53-5.57 (m, 4 nH, Ph-C¾-Im), 4.39-5.04 (m, 8 nH, -CH₂-C=C-CH₂-), 3.09-4.14 (m, 12 nH, DABCO N⁺- CH₂). The NMR data for copolymers IBN-131-1 and IBN-131-3 are the same as copolymer IBN-131-2.

IBN-132-3: ¾ NMR (400MHz, DMSO-d₆): δ 9.16-9.54 (m, 2 nH, Im C2H), 7.64-8.06 (m, 4 nH, Im C4H Im C5H , 7.33-7.49 (m, 4 nH, PhH, 6.93-7.12 (d, Im-C4H Im C5H terminal), 6.02-6.41 (m, 8 nH, -CH=CH-), 5.33-5.79 (m, 4 nH, Ph-CH-Im), 4.37-5.06 (m, 16 nH, -CH₂- C=C-CH-), 3.08-4.10 (m, 36 nH, DABCO N⁺-CH-). The NMR data for copolymers IBN-132- 1 and IBN-132-2 are the same as copolymer IBN-132-3.

For the second group of copolymers, tetramethylethane-l ,2-diamine (TMED) was utilized as the diamine, and copolymers IBN-211 and IBN-212 with para-xylylene linker were obtained using the same general procedure as described above, and based on the molar ratios and the respective diamine, di-imidazoles and aryl linkers as shown in the scheme below (Scheme 4). When ortho- xylylene or frares-butene linker was used, two small molecules (IBN-220 and IBN-230) were formed instead.

IBN-210 IBN-212

IBN-211

= p-phenylene; R = TMED; L1 , L2 = p-phenylene;

R = TMED; R = TMED; L1 , L2

X=Br

L1 = p-phenylene; X = Br L3 = p-phenylene; X=Br L3 = o-phenylene;

R:(L1+L2):L3 = 1 :2:1 R:(L1+L2):L3 = 1 :2:1

IBN-220 IBN-230

R = TMED; R = TMED;

L1 = o-phenylene; X L1 = frans-ethylene; X = Br

Scheme 4. Structures of TMED based polymer, small molecules, and TMED-imidazolium copolymers.

IBN-210: ¾ NMR (400MHz, DMSO-d₆): δ 7.71-7.97 (m, 4 nH, PhH), 4.89 (s, 4 nH, Ph-CH- TMED), 4.34 (s, 4 nH, N⁺-CH-CH-N⁺), 3.10-3.27 (d, 12 nH, N⁺-CH).

IBN-211: ¾ NMR (400Hz, DMSO-d₆): δ 9.51-9.65 (m, 2 nH, Im C2H), 6.94-7.87 (m, 16 nH, Im C4H Im C5H PhH), 5.20-5.58 (m, 8 nH, Ph-CH-Im), 4.89 (s, 4 nH, Ph-CH-TMED), 4.34 (s, 4 nH, N⁺-CH-CH-N⁺), 3.08-3.25 (d, 12 nH, N⁺-CH).

IBN-212: ¾ NMR (400Hz, DMSO-d₆): δ 9.56-9.59 (m, 2 nH, Im C2H), 6.90-7.98 (m, 16 nH, Im C4H, Im C5H PhH), 5.38-5.74 (m, 8 nH, Ph-CH-Im), 4.89 (s, 4 nH, Ph-CH-TMED), 4.34 (s, 4 nH, N⁺-CH-CH-N⁺), 3.09-3.27 (m, 12 nH, N⁺-CH).

As compared to DABCO and TMED, 1 ,4-dimethylpiperazine (DMP) is less reactive. Therefore, no polymer was formed when LI is o-phenylene. Polymers with >ara-xylylene and trans- butene linkers were obtained according to the general procedure as described above, and based on the molar ratios and the respective diamine, di-imidazoles and aryl linkers as shown in the scheme below (Scheme 5). However, their yields were lower than their analogues with DABCO or TMED diamines.

Scheme 5. Structures of DMP based polymers and ammonium-imidazolium copolymers.

IBN-310: ¾ NMR (400MHz, D₂0): δ 7.50-7.90 (m, 4 nH, F H), 4.85-5.03 (m, 4 nH, F -CH₂- DMP), 3.51-4.23 (m, 8 nH, N⁺-(¾-C¾-N⁺), 3.06-3.30 (m, 6 nH, N⁺-C¾).

IBN-330: ¾ NMR (400MHz, D₂0): δ 6.63 (d, 2 nH, , -CH=CH-), 4.52 (d, 4 nH, -CH₂-C=C- CH₂-), 3.81-4.34 (s, 8 nH, N⁺-C¾-C¾-N⁺), 3.19-3.40 (m, 6 nH, N⁺-C¾).

Example 3 - Molecular Weights of the Polymers and Copolymers

Molecular Weight Determination

A Waters 2695 gel permeation chromatography (GPC) equipped with two ultrahydrogel columns: 300 mm x 7.8 mm in series and a Waters 2414 differential refractometer detector (Massachusetts, U.S.A.) were used to determine the molecular weights of the polymers and copolymers. Details of the mobile phase used are as follows: 54/23/23 (v/v/v%) 0.5 M sodium acetate aqueous solution/methanol/acetic acid, at a flow rate of 1.0 mL/min. Calibration was performed using a series of poly(ethylene glycol) standards of varying molecular weights (633- 20,600) (Polymer Standard Service Inc., Rhode Island, U.S.A.). Mw and D was subsequently calculated from the calibration curve.

The molecular weights of the polymers and copolymers were measured with gel permeation chromatography (GPC). Similar to many step-growth polymers, these compounds are characterized by relatively low degree of polymerization (M_w< 10,000) and high dispersity values (1.3 < D <3.1) (Table 1). The distribution of molecular weights was affected by the polymers' solubility in solvents. Generally, the polymers have higher solubility in DMF than in THF due to the higher polarity of DMF. So most of the polymers synthesized in DMF have higher molecular weights.

Table 1. Molecular weight of polymers and copolymers.

Entry Polymer M_W(GPC) M„(GPC) M_w/M„

1 IBN-l lO(THF) 3066 2089 1.47

2 IBN-l l l(THF) 4920 2759 1.78 3 IBN-112(THF) 2844 1359 2.09

4 IBN-120(THF) 1311 994 1.32

5 IBN-121(THF) 2007 1216 1.65

6 IBN-122(THF) 1277 943 1.35

7 IBN-130(THF) 2340 1347 1.74

8 IBN-130(DMF) 2220 1573 1.41

9 IBN-131-1(THF) 3048 1508 2.02

10 IBN-131-2(THF) 2991 1449 2.06

11 IBN-131-3(THF) 2620 1331 1.97

12 IBN-132-1(THF) 4903 2094 2.34

13 IBN-132-2(THF) 3982 1845 2.16

14 IBN-132-3(THF) 2666 1270 2.10

15 IBN-132-1(DMF) 6260 2031 3.08

16 IBN-132-2(DMF) 7827 4622 1.69

17 IBN-132-3(DMF) 6002 3261 1.84

18 IBN-210(DMF) 6589 3453 1.91

19 IBN-211(DMF) 4638 2375 1.95

20 IBN-212(DMF) 4485 2442 1.84

21 IBN-310(DMF) 3442 1703 2.02

22 IBN-311(DMF) 5993 2984 2.01

23 IBN-312(DMF) 5484 2593 2.12

24 IBN-330(DMF) 5604 2254 2.49

25 IBN-331(DMF) 9463 6893 1.37

26 IBN-332(DMF) 8394 3787 2.22

Example 4 - Antimicrobial and Antifungal Studies

Minimum Inhibitory Concentration

Staphylococcus aureus (ATCC 6538, Gram-positive), Escherichia coli (ATCC 8739, Gram- negative), Pseudomonas aeruginosa (ATCC 9027, Gram-negative), and Candida albicans (ATCC 10231, fungus) were used as representative microorganisms to challenge the antimicrobial functions of the imidazolium salts. All bacteria and fungus were stored frozen at - 80 °C, and were grown overnight at 37 °C in Mueller Hinton Broth (MHB, BD Singapore) prior to experiments. Fungus was grown overnight at 22 °C in Yeast Mold broth (YMB, BD Singapore). Subsamples of these cultures were grown for a further 3 hours and diluted to give an optical density (O.D.) value of 0.07 at 600 nm, corresponding to 3 xlO⁸ CFU mL ¹ for bacteria and 10⁶ CFU mL ¹ for fungus (McFarland's Standard 1; confirmed by plate counts).

The polymers were dissolved in MHB or YMB at a concentration of 4 mg mL ¹ and the minimal inhibitory concentrations (MICs) were determined by microdilution assay. Bacterial solutions (100 μL, 3 χ 10⁸ CFU mL ¹) were mixed with 100 μΐ_^ of polymer solutions (normally ranging from 4 mg mL ¹ to 2 μg mL ¹ in serial two-fold dilutions) in each well of the 96-well plate. The plates were incubated at 37 °C for 24 hours with constant shaking speed at 300 rpm. The MIC measurement against Candida albicans is similar to bacteria except that the fungus solution is ~10⁶ CFU mL ¹ in YMB and the plates were incubated at room temperature.

The minimum inhibitory concentrations were taken as the concentration of the antimicrobial oligomer/polymer at which less than 50% microbial growth was observed with the microplate reader (TEC AN). Medium solution containing microbial cells alone were used as control (100% microbial growth). The assay was performed in four replicates and the experiments were repeated at least two times.

Antifungal Activity and Minimum Fungicidal Concentration (MFC)

IBN-132-3 (THF) was dissolved in YMB (2 μg/mL to 62 μg/mL in serial two-fold dilution). A hundred microliters of each solution were placed into a 96-well microplate. A hundred microliters of C. albicans suspension (10⁶ CFU/ml) was then added into each well. Fungus growing in YMB was used as control. For MFC, antibiotics, Amphotericin B and Fluconazole, were also tested as positive control. The 96-well plate was kept in an incubator at room temperature under constant shaking. After incubation for desired period, the respective cell suspensions were collected (100 μί), serially diluted 1: 10, and 100 μΐ_^ of each dilution was spread on two nutrient agar plates (Luria-Bertani broth with 1.5% agar). Colony forming units (CFU) were counted after 48 hours incubation and the CFU/mL was calculated accordingly. Table 2. Antimicrobial activity of controls under conditions in this study.

MIC ^g/ml)

Entry Antimicrobials

E. coli S. aureus P. aeruginosa C. albicans

1 Chlorhexidine 16 - 31 4

2 Vancomycin - 2 - -

3 Amphotericin B - - - 4

4 Fluconazole - - - >500

Benzalkonium

5 31 31 63 16

chloride 6 Chloroxylenol > 100 > 100 >100 >100

Resistance Studies

Drug resistance was induced by treating S. aureus or C. albicans repeatedly with copolymers and control antibiotics. First MICs of the tested copolymers were determined against S. aureus or C. albicans using the broth microdilution method. Then serial passaging was initiated by transferring microbial suspension grown at the sub-MIC of the copolymers (1/2 of MIC at that passage for S. aureus and 1/8 of MIC at that passage for C. albicans) for another MIC assay. After 24 hours of incubation, cells grown at the sub-MIC of the test compounds/antibiotics were once again transferred and assayed for MIC. The MIC against S. aureus or C. albicans was tested for 15 passages.

Drug-resistant behavior was evaluated by recording the changes in the MIC normalized to that of the first passage. Conventional antibiotic Amphotericin B was used as the control against C. albicans and Norfloxacin was used as the control against S. aureus.

Preparation of Biofilm in vitro

Fungus C. albicans cells were cultured in YMB at room temperature. Bacteria cells were cultured in MHB at 37 °C. All the microorganisms were grown overnight to reach mid logarithmic growth phase. The concentrations of the microbe were adjusted to give an O.D. value of 0.07 at 600 nm. The bacterial solutions were then diluted 10³ fold to achieve an initial loading of 3 x 10⁵ CFU/mL while C. albicans cells were used without further dilution. 100 μΐ_^ of the microbial solution was added to each well of 96-flat bottom well plate and incubated at room temperature for the fungus and at 37 °C for the bacteria, under constant shaking. After 24 hours, the media in each well was discarded and 100 μΐ_^ of 3 χ 10⁵ CFU/mL fresh microbial suspension was added to the wells. This was repeated every day for 7 days to allow biofilm formation. For C. albicans, after 24 hours, fresh medium without fungus was added to replace the microbial solution. The incubation went on for 2 more days. After 7 days or 3 days of incubation, the biofilm formed in wells was rinsed three times with phosphate -buffered saline (PBS), before further analysis.

Biofilm Susceptibility Test Using XTT Reduction Assay

A (2-methoxy-4-nitro-5-sulfo-phenol)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay was used to quantify the live microbe on the surface of each well by measuring the mitochondrial enzyme activity in live cells. In this assay, mitochondrial dehydrogenases of the viable microbial cells reduced XTT to an orange colored formazan derivative, and the change in O.D. reading was recorded to analyze the viability of cells on the surfaces. The polymer/copolymer solutions (100 μί-, different concentrations) were added to the wells containing biofilm. Antibiotic solutions and pure medium were used as controls. After 24 hours of incubation, the wells were washed with PBS once and then submerged in a mixture solution of 100 μί- PBS, 10 μΐ_^ XTT (1 mg/mL), and 2 μί menadione (0.4 mM). After incubation at 37 °C for 4 hours, the absorbance of the samples at 490 nm was measured using a microplate reader with 600 nm as reference wavelength. Relative cell viability was calculated using the formula, [(OD490nm-OD6oonm)poiymer]/[(OD₄90nm-OD6oonm)controi] lOO . Data were represented as means + standard deviation of four replicates for each concentration. Haemolysis Study

Fresh rat red blood cells (RBCs) were diluted with PBS buffer to give an RBC stock suspension (4 vol% blood cells). 100 μί. aliquots of RBC suspension were mixed with 100 copolymer solutions of various concentrations (ranging from 4 mg mL ¹ to 2 μg mL ¹ in serial two-fold dilutions in PBS). After 1 hour of incubation at 37 °C, the mixture was centrifuged at 2000 rpm for 5 min. Aliquots (100 μί) of the supernatant were transferred to a 96-well plate. Haemolytic activity was determined as a function of hemoglobin release by measuring absorbance of the supernatant at 576 nm using a microplate reader. A control solution that contained only PBS was used as a reference for 0% haemolysis. Absorbance of red blood cells lysed with 0.5% Triton-X was taken as 100% haemolysis. The data were expressed as mean and S.D. of four replicates, and the tests were repeated two times.

% HaemolySlS [OD576nm (polymer)-ODs76nm (PBS)]/[OD576nm (Triton X)-OD576nm (PBS)]xl00%

Scanning Electron Microscopy (SEM) Observation

C. albicans cells (10⁶ CFU/mL) grown in YMB without or with the copolymers at 125

for 24 hours were collected and centrifuged at 3000 rpm for 5 minutes. The precipitates were washed twice with PBS buffer. Then the samples were fixed with glutaraldehyde (2.5%) for 4 hours followed by washing with deionized (DI) water. Dehydration was performed using a series of ethanol/water solution (35%, 50%c, 75%o, 90%o, 95%o and 100%). The dehydrated samples were mounted on copper tape. After drying for 2 days, the samples were further coated with platinum for imaging with JEOL JSM-7400F (Japan) field emission scanning electron microscope operated at an accelerating voltage of 3 keV.

Statistical Analysis

Data were expressed as means + standard deviation of the mean (S.D. is indicated by error bars). Student's i-test was used to determine significance among groups. A difference with p<0.05 was considered statistically significant.

Example 5 - DABCO-Imidazolium Copolymers

Antimicrobial Activity of DABCO-Imidazolium Copolymers

The antimicrobial activities of DABCO-imidazolium copolymers were evaluated against four different and clinically relevant microbes: S. aureus, E. coli, P. aeruginosa, and C. albicans. Their minimum inhibitory concentrations (MICs) against the four microbes were presented in Table 3. All the DABCO polymers and DABCO-imidazolium copolymers exhibit antimicrobial activities against the tested microbe. Interestingly, the structure of the linkers affects the antimicrobial activity. In general, polymer and copoplymers containing rrares-butenyl linkers are the most active material. Polymer and copoplymers with o-xylenyl linkers are more active than those with />-xylenyl linkers. Therefore, the antimicrobial activity sequence of polymers is of fraras-butene linker > orf/zo-xylylene linker > ara-xylylene linker.

The DABCO-imidazolium copolymers with iraras-butene linker show superior antifungal activity (Table 3). MICs against C. albicans are all less than 10 μg/mL. They are more effective than DABCO polymer IBN-130 or imidazolium polymer PIM-45, implying synergistic effect when DABCO and imidazolium are combined together. Copolymers synthesized in THF, which generally have lower molecular weight than polymer synthesized in DMF, demonstrate higher antifungal activity and lower toxicity (Table 3, entries 15 to 20). Particularly, IBN-132-3, synthesized in THF with DABCO to di-imidazolium ratio is 3, is the most active antifungal compound with MIC of 2 μg/mL. No significant hemolysis was observed at the highest concentration, 2000 μg/mL of IBN-132-3.

Table 3. Minimum inhibitory concentrations (MICs) of DABCO based polymers and copolymers."

Compound MICso ^g/ml) HCio

Entry Mw

IBN- E. C. S. A. P. A. C. A. F. S. ^g/ml)

1 110 (THF) 3066 > 125 62 62 125 - >2000

2 110 (DMF) - >62 16 62 62 - >2000

3 111 (THF) 4920 31 16 31 62 - 62

4 111 (DMF) - 31 16 31 62 62 62

5 112 (THF) 2844 16 16 31 62 - 250

6 112 (DMF) - 31 16 62 62 125 >2000

7 120 (THF) 1311 > 125(125^b) 62(31 ^b) 62 31 - >2000

8 121 (THF) 2007 31(8 ^b) 16(8 ^b) 31 16 - >2000

9 122 (THF) 1277 31(8 ^b) 16(8 ^b) 62 31 - >2000

10 130 (THF) 2340 31 16 31 8 62 >2000

11 130 (DMF) 2220 62 31 62 8 125 >2000

12 131-1 (THF) 3048 16 16 31 8 - 2000

13 131-2 (THF) 2991 16 8 62 4 - >2000

14 131-3 (THF) 2620 31 16 62 4 - 2000

15 132-1 (THF) 4903 16 8 16 8 31 >2000

16 132-1 (DMF) 6260 16 16 16 16 - 1000

17 132-2 (THF) 3982 16 8 31 4 31 >2000

18 132-2 (DMF) 7827 31 16 16 16 - 62

19 132-3 (THF) 2666 16 16 62 2 31 >2000

20 132-3 (DMF) 6002 31 16 31 16 - 62

21 PIM-45 2000 8 8 31 31 125 >2000

22 Amphotericin B - - - 4 125 >2000 ^a MIC tested against ~ 3xl0⁸ CFU/mL bacteria or 10⁶ CFU/mL fungi. E. C. (E. coli), S. A. (S. aureus), P. A. (P. aeruginosa), C. A. (C. albicans), F. S. (F. solani).

b MIC tested against 10^s CFU/mL bacteria.

The killing efficacy of IBN-132-3 (THF) at different concentrations against C. albicans after 48 hours of treatment was shown in Fig. 1A. IBN-132-3 can inhibit the growth of C. albicans at low concentration, even below MIC. At half MIC (1 μg/mL), the growth of C. albicans is slower than control. The inhibition increases when the concentration increases and 3 log reduction was observed at 8 μg/mL concentration. The antifungal activity of IBN-132-3 was compared with two conventional antibiotics which are currently used in clinical treatment, Amphotericin B (AmB) and Fluconazole (Fig. IB and Table 4). The growth of C. albicans in the presence of Fluconazole has no significant difference with untreated control, which means Fluconazole is inactive against our tested strain. The MIC of IBN-132-3 is 2 μg/mL, lower than that of AmB (4 μg mL for 24 hours treatment). Although the minimum fungicidal concentration (MFC) of AmB for 24 hours treatment (16 μg/mL) is lower than that of IBN-132-3 (62 μg/mL), after 48 hours, the concentration of C. albicans increased due to amphotericin B's poor stability.

Table 4. Comparison of MICs and MFCs between IBN-132-3 and antibiotics.

MICso (24 h) MICso (48 h) MFC (24 h) MFC (48 h)

IBN-132-3 2 2 62 62

Amphotericin B 4 16 16 > 16

Fluconazole >500 >500 >500 >500

Example 6 - TMED-Imidazolium and DMP-Imidazolium Copolymers

Antimicrobial Activity of TMED-Imidazolium Copolymers and DMP-Imidazolium Copolymers

The MICs of copolymers with TMED or DMP units were also tested and the results were shown in Table 5 and 6. The polymers and copolymers obtained all show good antimicrobial activity. The two small molecules, IBN-220 and IBN-230 are inactive.

Table 5. Minimum inhibitory concentrations (MICs) of TMED based polymers and copolymers."

MICso (pg/ml)

Entry Compound IBN- Mw HQo ^g/mL)

E. C. S. A. P. A. C. A. F. S.

1 210 (THF) - 31(8^b) 16(4^b) 31 31 62 >2000

2 210 (DMF) 6589 31 16 31 31 62 >2000

3 211 (THF) - 31 16 31 31 125 125

4 211 (DMF) 4638 16 16 31 31 62 125

5 212 (THF) - 16 16 31 31 - 2000

6 212 (DMF) 4485 16 8 31(16^b) 16 31 2000

7 220 >1000 >1000 - - - -

8 230 >1000 >1000 >1000 >2000 - - ^a MIC tested against -3x10^s CFU/mL of bacteria or 10^b CFU/mL of fungi. E. C. (E. coli), S. A. (S. aureus), P. A. (P. aeruginosa), C. A. (C. albicans), F. S. (F. solani).

b MIC tested against 10^s CFU/mL of bacteria. Table 6. Minimum inhibitory concentrations (MIC) of DMP based polymers/copolymers.

Entry Compound IBN- Mw HC₁₀ (Mg/mL)

E. C. S. A. P. A. C. A.

1 310 (THF) - 31(31) 16(8) 31(16^b) 8 >2000

2 310 (DMF) 3442 31 16 31 16 >2000

3 311 (DMF) 5993 16 8 16 8 >2000

4 312 (DMF) 5484 16 8 16 8 >2000

5 330 (THF) - 31(31) 16(8) 62(16) 4 >2000

6 330 (DMF) 5604 31 16 125 4 1000

7 331 (DMF) 9463 31 16 31 8 62

8 332 (DMF) 8394 31 16 31 8 >2000 ^a MIC tested against ~3xl0⁸ CFU/mL of bacteria or 10⁶ CFU/mL of fungi. E. C. (E. coli), S. A.

(S. aureus), P. A. (P. aeruginosa), C. A. (C. albicans), F. S. (F. solani).

b MIC tested against 10^s CFU/mL of bacteria.

Example 7

Drug Resistance Study

The potential for bacterial/fungal cells to develop resistance following repeated exposures to copolymers was investigated by serial passage of S. aureus or C. albicans treated under sub- MIC levels for each polymer. MIC values were measured after each passage. For comparison, antibiotics Norfloxacin and Amphotericin B, were also tested. As shown in Fig. 2 A, the MIC of Norfloxacin against S. aureus increased at the 4^th passage. By the 15^th passage, the MIC of Norfloxacin increased to 124 times of the original MIC. In contrast, the MICs of IBN-131-2, IBN-132-3 and IBN-212 (DMF) remained unchanged over the entire 15 passages. The results indicated the much lower propensity of bacteria to develop resistance toward the DABCO- imidazolium copolymers and TMED-imidazolium copolymers compared to Norfloxacin.

Resistance to antifungal agents has been much less studied than antibacterial resistance. However, the current increase in fungal infections has intensified the exploration of innovative, safer and more efficient agents to combat fungal infections. As shown in Fig. 2B, the MICs of IBN-131-2, IBN-132-3 and IBN-212 against C. albicans remained constant over 15 passages of sub-MIC dose treatment, indicating that C. albicans cannot develop drug resistance against the copolymers in the 15 consecutive days of treatment. The MIC of Amphotericin B increased at the 4^lh passage and quadrupled at the 5^th passage, suggesting that resistance can be induced when fungi were treated with Amphotericin B repeatedly.

Scanning Electron Microscopy (SEM) Observation

The antifungal mechanism was studied by visualizing a typical cell structure with or without treatment via scanning electron microscopy (SEM). The morphological changes of C. albicans after being treated with copolymers are shown in Fig. 3. Compared with the intact cell wall of the control (Fig. 3A), the cell wall of copolymer -treated C. albicans was disrupted and subsequently dissolved after 24 hours of exposure (Fig. 3B to Fig. 3D). It is hypothesized that the copolymers work via an associative mechanism, which required appropriately balanced hydrophobic and hydrophilic regions to kill fungi. The copolymers became integrated within the cellular exterior causing membrane destabilization and lysis. This membrane -lytic mechanism might be the reason for reduced potency of developing drug resistance.

Biofilm Susceptibility

Individual organisms in biofilms are embedded within a matrix of slimy, extracellular polymers and typically display a phenotype that is very different from that of planktonic cells. In particular, bacteria/fungi in biofilms are much more tolerant to antimicrobials than their planktonic counterparts. As a result, drug treatments for biofilms are sometimes futile. It was reported that Candida biofilms are resistant to several clinically important antifungal agents, including Amphotericin B and Fluconazole. Herein, the activities of the copolymers against biofilms were also tested.

As shown in Fig. 4, all the tested copolymers demonstrated strong antifungal and antimicrobial activity against C. albicans (Fig. 4A) biofilm and three bacterial [E.coli (Fig. 4B), S. aureus (Fig. 4C) and P. aeruginosa (Fig. 4D)] biofilms. For example, 80% of C. albicans was eradicated in 24 hours after a single treatment with IBN-132-3 at 31 g/mL. In contrast, C. albicans treated with the same concentration of Amphotericin B did not show significant difference compared to non-treated control. Similarly, 87% of E. coli (Fig. 4B) was eradicated in 24 hours after a single treatment with IBN-212 at 125 μg mL. The results demonstrated that the copolymers were effective against biofilms.

The above examples demonstrated that these ammonium-imidazolium copolymers possess excellent antimicrobial activity against a broad range of microbe or microorganism and related biofilms, and may possess essential degradation and non-resistance properties. Ammonium- imidazolium copolymers comprising iraras-butenyl linkers may be the most active material. The activity of the ammonium-imidazolium copolymers can be adjusted by using different ratio of monomers and linkers with different hydrophobicity and flexibility properties.

Industrial Applicability

The polymers as defined above may be used as a composition in association with a carrier or a pharmaceutical composition in association with a pharmaceutically acceptable carrier. The polymer or the composition as defined above may be used as a non-therapeutic agent for killing or inhibiting the growth of a microorganism. The polymers as defined above may be used in a number of applications due to their ability to inhibit the growth of a microorganism.

The polymers or the pharmaceutical composition as defined above may be used in a number of applications in due to their ability to inhibit the growth of a microorganism or be used in treating a microbial infection or disease whereby the polymer or the pharmaceutical composition is administered to a subject. The polymers or the pharmaceutical composition as defined above may be used for killing or inhibiting the growth of a microorganism. The polymers or the pharmaceutical composition as defined above may be used in the manufacture of a medicament for killing or inhibiting the growth of a microorganism. The polymers or the pharmaceutical composition as defined above may be used for treating a microbial infection or disease. The polymers or the pharmaceutical composition as defined above may also be used as an antibiotic. The polymers or the pharmaceutical composition as defined above may be used in the manufacture of a medicament for treating a microbial infection or disease. The microbial infection or disease may be caused by a microorganism that is selected from the group consisting of bacterium, archaea, fungus, protist, animal, plant, or any mixture thereof.

The polymers as defined above may exhibit antimicrobial activities against the tested microbe (bacterium) e.g. Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, and. The polymers as defined above may exhibit antifungal activities against the fungal species e.g. Candida albicans and Fusarium solani.

The new ammonium-imidazolium copolymers as defined above may have tuneable degradation profiles under different conditions, which would have wide ranging applications in agricultural and environmental disinfection. Bacteria and fungi may have showed lower propensity to develop resistance toward the copolymers compared to conventional antibiotics. Among them, DABCO-imidazolium copolymers exhibited excellent antifungal activity and biocompatibility. Advantageously, these copolymers are easy to synthesize and relatively low cost, with wide ranging applications in medical, agricultural and environmental disinfection. The copolymers may also be used in topical wound treatment, as preservatives or disinfectants for consumer care and personal care products.

It will be apparent that various other modifications and adaptations of the invention will be apparent to the person skilled in the art after reading the foregoing disclosure without departing from the spirit and scope of the invention and it is intended that all such modifications and adaptations come within the scope of the appended claims.

Claims

1. A polymer having the following Formula (I):

Formula (I)

wherein

X is independently selected from a halogen;

n, m and p are independently an integer of at least 1 ;

q is 0 or an integer of at least 1 ;

A has the following structure:

x and y are independently an integer of at least 1 ,

or a salt or hydrate thereof.

2. The polymer according to claim 1, wherein n and m are independently an integer between 1 and 50.

3. The polymer according to claim 1, wherein x and y are independently an integer between 1 and 5.

4. The polymer according to claim 3, wherein x and y are independently 1.

5. The polymer according to any one of the preceding claims, wherein any two of R¹, R², R³ and R⁴ is taken together to form at least one C₂ to C₅ bridging group.

6. The polymer according to claim 5, wherein R¹ and R² are taken together to form an ethenylene group.

7. The polymer according to claim 6, wherein R³ and R⁴ are taken together to form an ethenylene group.

8. The polymer according to any one of claims 5 to 7, wherein A is selected from the group consisting of the following structures:

9. The polymer according to any one of the preceding claims, wherein the molar ratio between A and L¹ is in the range of 1 :5 and 5: 1.

10. The polymer according to any one of the preceding claims, wherein the molar ratio between (L¹ + L²) and L³ is in the range of about 1 :5 to about 5:1.

11. The polymer according to any one of the preceding claims, wherein the polymer has a molecular weight in the range of about 1000 to about 10,000.

12, The polymer according to any one of the preceding claims, wherein the polymer has a polydispersity value in the range of about 1.2 to about 3.2.

13, The polymer according to any one of the preceding claims, having the following Formula (II):

Formula (II)

wherein

L⁴ and L⁵ are independently selected from optionally substituted alkenyl or optionally substituted aryl.

14, The polymer according to claim 13, wherein L⁴ and L⁵ are independently selected from the group consisting of ort/xo-phenylene group, para-phenylene group, meto-phenylene group and ethenylene.

The polymer according to any one of the preceding claims, wherein X is CI or Br.

16. The polymer according to any one of claims 13 to 15, wherein the following portion in

formula (II):

selected from the group consisting of the following structures:

17. The polymer according to any one of the preceding claims, wherein p and q are independently an integer between 1 and 5.

18. The polymer according to claim 17, wherein p and q are independently 1.

19. The polymer according to any one of the preceding claims, selected from the group consisting of:

20. A composition comprising the polymer according to any one of the preceding claims, or a salt or hydrate thereof, in association with a carrier.

21. Use of the polymer according to any one of claims 1 to 19, or the composition according to claim 20, as a non -therapeutic agent for killing or inhibiting the growth of a microorganism.

22. A pharmaceutical composition comprising the polymer according to any one of claims 1 to 19, or a pharmaceutically acceptable salt or hydrate thereof, in association with a pharmaceutically acceptable carrier.

23. A method for killing or inhibiting the growth of a microorganism, the method comprising administering to a subject the polymer of any one of claims 1 to 19, or the pharmaceutical composition of claim 22.

24. A polymer according to any one of claims 1 to 19, or a pharmaceutical composition according to claim 22, for killing or inhibiting the growth of a microorganism.

25. Use of the polymer according to any one of claims 1 to 19, or the pharmaceutical composition according to claim 22, in the manufacture of a medicament for killing or inhibiting the growth of a microorganism.

26. The method, polymer, pharmaceutical composition or use according to any one of claims 21 or 23 to 25, wherein the microorganism is a bacterium, archaea, fungus, protist, animal, plant, or any mixture thereof.

27. A method for treating a microbial infection, the method comprising administering to a subject the polymer of any one of claims 1 to 19, or the pharmaceutical composition of claim

22.

28. A polymer according to any one of claims 1 to 19, or a pharmaceutical composition according to claim 22 for use as an antibiotic.

29. Use of the polymer according to any one of claims 1 to 19, or the pharmaceutical composition according to claim 22, in the manufacture of a medicament for treating a microbial infection.

30. The method, polymer, pharmaceutical composition or use according to any one of claims 27 to 29, wherein the microbial infection is caused by a bacterium, archaea, fungus, protist, animal, plant, or any mixture thereof.

31. A method for preparing the polymer according to any one of claims 13 to 19, comprising the steps of:

contacting a diamine having the following structure:

, wherein R¹, R², R³ and R⁴ are independently selected from an optionally substituted alkyl, or any two of R¹, R², R³ and R⁴ is taken together to form at least one bridging group; and x is an integer of at least 1 ;

with a di-imidazole having the following structure:

, wherein L is selected from the group consisting of or?¾o-phenylene group, /?ara-phenylene group, meto-phenylene group and ethenylene;

and a dihalide having the following structure:

, wherein L⁵ is selected from the group consisting of ortho- phenylene group, para-phenylene group, meia-phenylene group and ethenylene; and X is a halide;

under reaction conditions.

32. The method according to claim 31, wherein the diamine and di-imidazole are contacted at a molar ratio in the range of about 1 :5 to about 5: 1.

33. The method according to claim 31, wherein di-imidazole and dihalide are contacted at a molar ratio in the range of about 1:6 to about 5:6.

34. The method according to any one of claims 31 to 33, wherein the reaction is performed in a polar organic solvent, wherein the polar solvent is tetrahydrofuran (THF), dimethylformamide (DMF) or any mixture thereof.

35. The method according to any one of claims 31 to 34, wherein the reaction is carried out at a temperature in the range of about 60 °C to about 100 °C.

36. The method according to any one of claims 31 to 35, wherein the reaction is carried out for at least 18 hours.