WO2024107810A1 - Phenyl-free biscationic quaternary phosphonium compounds as antimicrobial agents - Google Patents

Phenyl-free biscationic quaternary phosphonium compounds as antimicrobial agents Download PDF

Info

Publication number
WO2024107810A1
WO2024107810A1 PCT/US2023/079774 US2023079774W WO2024107810A1 WO 2024107810 A1 WO2024107810 A1 WO 2024107810A1 US 2023079774 W US2023079774 W US 2023079774W WO 2024107810 A1 WO2024107810 A1 WO 2024107810A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
alkyl
vial
antimicrobial composition
nmr
Prior art date
Application number
PCT/US2023/079774
Other languages
French (fr)
Inventor
Kevin P.C. Minbiole
William WUEST
Original Assignee
Villanova University
Emory University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Villanova University, Emory University filed Critical Villanova University
Publication of WO2024107810A1 publication Critical patent/WO2024107810A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/54—Quaternary phosphonium compounds
    • C07F9/5449—Polyphosphonium compounds
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
    • A01N57/18—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-carbon bonds
    • A01N57/20—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-carbon bonds containing acyclic or cycloaliphatic radicals
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof

Definitions

  • the disclosure relates to antimicrobial compositions and related methods generally. More particularly, the disclosed subject matter relates to a biscationic quaternary phosphonium compound, a composition comprising the biscationic quaternary phosphonium compound, the methods of making the same, and the methods of using the same as antimicrobial agents.
  • quaternary ammonium compounds have been employed as successful tools to mitigate the transfer of pathogenic bacteria.
  • society has heavily relied on just a handful of these disinfecting amphiphilic compounds, with the most visible commercial disinfectant of this class being benzalkonium chloride (BAC), a mixture of long-chained alkyl dimethylbenzylammonium chlorides.
  • BAC benzalkonium chloride
  • mainstay disinfectants such as BAC have proven highly valuable for decades, they are now showing vulnerabilities as bacteria continue to evolve antimicrobial resistance mechanisms.
  • Notable pathogens such as A. baumarmii and P. aeruginosa have developed sufficient resistance mechanisms towards common disinfectants that they are renewing the problem of bacterial infections in hospital, domestic and commercial settings.
  • the present disclosure provides a biscationic quaternary phosphonium compound, an antimicrobial composition comprising such a compound, the method of making such a compound, the method of making such an antimicrobial composition, and the methods of using such a compound or composition for antimicrobial use.
  • the biscationic quaternary phosphonium compound is phenyl-free in some embodiments.
  • the compound or the composition provided in the disclosure has an ability to kill or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, to attenuate the severity of a microbial infection, or to prevent or inhibit formation of a biofilm or eradicate pre-established biofilms (i.e. antibiofilm use).
  • the present disclosure provides an antimicrobial composition
  • L is a linking group being C 1-10 alkyl or a cycloalkyl
  • R 1 , R 2 , R 3 , or R 4 each is a C 1-12 alkyl or a cycloalkyl, unsubstituted or optional substituted with a functional group selected from the group consisting of -OH, -OR’, -NH 2 , - NHR’, -NR’2, -SH, -SR’, -O-C(O)R’, -C(O)OR’, -C(O)R’, -CF 3 , -OCF 3 , halogen, and any combination thereof;
  • R’ is H or a C 1-4 alkyl
  • X is selected from the group consisting of a halogen, a tosylate, a hydrocarbonate, a carbonate, a sulfate, and an acetate
  • m is 1 or 2.
  • the present disclosure provides a composition or a product described herein.
  • the antimicrobial composition is a disinfectant configured to be applied to a surface of an object in need thereof.
  • the antimicrobial composition is a pharmaceutical composition configured to be administrated to a subject in need thereof.
  • the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an active ingredient.
  • the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an additive for stabilizing the pharmaceutical composition.
  • the present disclosure provides a compound as described herein, a method of making such a compound, and a method of using such a compound.
  • the present disclosure provides a method of making the antimicrobial composition as described herein.
  • such a method comprises preparing the compound having the formula (I) as described. The method may further comprise mixing an effective amount of the compound having the formula (I) and a carrier.
  • the present disclosure also provides different products as described herein, which comprise the antimicrobial composition.
  • the present disclosure provides a method of killing, preventing, or inhibiting microbial growth.
  • the method comprises applying the antimicrobial composition to a surface of an object in need thereof.
  • such a method comprises administrating the antimicrobial composition to a subject in need thereof.
  • the antimicrobial composition or the compound is used to kill, prevent, or inhibit growth of at least one group of microorganisms such as bacteria, viruses, yeast, fungi, and protozoa, or to inhibit formation of a biofilm, or disperse or eradicate a pre-established biofilm.
  • FIG. 1 shows characterization data for structurally analogous bisQAC and bisQPC compounds 12(2)12 and MeP2P-12,12 with bromide counter ions. Shown are dynamic surface tension (mN/rn) data when plotted as a function of surface age (ms) at three concentrations of 100, 300 and 1000 ppm.
  • mN/rn dynamic surface tension
  • the recited range may be construed as including situations whereby any of 1, 2, 3, 4, or 5 are negatively excluded; thus, a recitation of “1 to 5" may be construed as “1 and 3-5, but not 2”, or simply “wherein 2 is not included.” It is intended that any component, element, attribute, or step that is positively recited herein may be explicitly excluded in the claims, whether such components, elements, attributes, or steps are listed as alternatives or whether they are recited in isolation.
  • antimicrobial refers to an ability to kill, prevent, or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, or to attenuate the severity of a microbial infection.
  • the antimicrobial compounds or compositions of the present invention are compounds or compositions that may be used for cleaning or sterilization, or may be used in the treatment of disease and infection.
  • the applications may include both in vitro and in vivo antimicrobial uses.
  • Applying” an antimicrobial composition may include administrating a composition into a human or animal subject.
  • biofilm refers to a film formed by a group of microorganisms adhered together.
  • antibiofilm refers to an ability to kill, disperse and/or eradicate a pre-established biofilm.
  • alkyl refers to a straight chain, cyclic, branched or unbranched saturated or unsaturated hydrocarbon chain containing 1-25 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n- septyl, n-octyl, n-nonyl, cyclo-hexyl, and the like.
  • the alkyl may be straight or branched, and can be optionally substituted.
  • a C 1-12 alkyl refers to an alkyl group having a number of carbon atoms selected from 1 to 12. Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (referred to as an “alkenyl” or “alkynyl,” respectively).
  • Representative straight chain and branched alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl, 1 -pentenyl, 2-pentenyl, 3 -methyl- 1-butenyl, 2-methyl-2- butenyl, 2,3- dimethyl-2-butenyl, and the like; while representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1 -pentynyl, 2-pentynyl, 3- methyl- 1-butynyl, and the like.
  • optionally substituted means that group in question may be unsubstituted or it may be substituted one or several times, such as 1 to 3 times or 1 to 5 times.
  • an alkyl group that is “optionally substituted” with 1 to 5 chloro atoms may be unsubstituted, or it may contain 1, 2, 3, 4, or 5 chlorine atoms.
  • Substituted chemical moieties include one or more substituents that replace hydrogen.
  • microorganism refers to a small (often, but not always, microscopic) organism that is typically, but not exclusively, single cellular, and includes organisms from the kingdoms bacteria, archaea, protozoa, and fungi.
  • the present disclosure is primarily directed to microorganisms that are pathogenic and capable of causing disease. Such microbes may be detected from a sample.
  • microorganism includes bacteria and fungi capable of causing disease, particularly disease in humans and other mammals and animals in need of treatment.
  • object refers to any article having a solid surface.
  • An object may also include a body part of a subject.
  • a composition to be applied to a surface of an object may be in a form of liquid or aerosol.
  • a “subject” refers any animal, preferably a human being such as a patient, livestock, rodent, monkey, or domestic pet. In some embodiments, the subject may be exhibiting symptoms of, at risk of, or diagnosed with a disease or condition by analysis of a sample.
  • sample can refer to a tissue sample, cell sample, a fluid sample, and the like.
  • a sample may be taken from a host subject
  • the tissue sample can include hair, buccal swabs, blood, saliva, semen, muscle, or tissue from any internal organ.
  • the fluid may be, but is not limited to, urine, blood, ascites, pleural fluid, spinal fluid, semen, wound exudates, sputum, fecal matter, saliva, and the like.
  • the body tissue can include, but is not limited to, skin, muscle, endometrial, uterine, and cervical tissue.
  • a sample in the context of the present disclosure, is primarily a biological sample (e.g., from a living host) the sample may also be an environmental sample suspected of contamination by microbes, such as a water sample, food sample, soil sample, and the like.
  • a liquid sample and some solid samples may be used as a test sample without modification for testing directly, if a solid sample is to be made into liquid form for testing and/or a liquid sample is to be diluted, a test sample may be made by reconstituting, dissolving, or diluting the sample in a fluid such as water, buffered saline, and the like.
  • the terms “administration” or “administrating” is understood to encompass any type of application to a subject including in vitro or in vivo administrations.
  • the types of administration include, are not limited to, topical, oral, parenteral, rectal, intramuscular injection, inhalation, intrathecal, sublingual, nasal, transdermal, and any combination thereof.
  • the composition to be administrated may be in a form of a solid dose form such as a tablet or a capsule, a liquid form, or an aerosol.
  • the terms “prevent” and “preventing” include the prevention of the recurrence, spread or onset. It is not intended that the present disclosure be limited to complete prevention. In some embodiments, the onset is delayed, or the severity is reduced. [0028] As used herein, the terms “treat” and “treating” are not limited to the case where the subject (e.g. patient) is cured and the disease is eradicated. Rather, embodiments of the present disclosure also contemplate treatment that merely reduces symptoms, and/or delays disease progression.
  • ⁇ ективное amount refers to that amount of a compound or pharmaceutical composition described herein that is sufficient to effect the intended application including, but not limited to, disease treatment, as illustrated below.
  • the therapeutically effective amount can vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art.
  • the term also applies to a dose that will induce a particular response in target cells, e.g., reduction of platelet adhesion and/or cell migration.
  • the specific dose will vary depending on, for example, the particular compounds chosen, the dosing regimen to be followed, whether it is administered in combination with other agents, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.
  • linking group refers to any variety of molecular arrangements that can be used to bridge molecular moieties together.
  • this disclosure contemplates derivatives of compounds disclosed herein.
  • the term “derivative” refers to a structurally similar compound that retains sufficient functional attributes of the identified analogue.
  • the derivative may be structurally similar because it is lacking one or more atoms, substituted, a salt, or alternative salt, an ester, in different hydration/oxidation states, or because one or more atoms within the molecule are switched, such as, but not limited to, replacing an oxygen atom with a sulfur atom, or replacing an amino group with a hydroxy group.
  • substituted refers to a molecule wherein at least one hydrogen atom is replaced with a substituent When substituted, one or more of the groups are “substituents.”
  • Example substituents may include halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, amine, and ester.
  • cell culture or “growth medium” or “media” refers to a composition that contains components that facilitate cell maintenance and growth through protein biosynthesis, such as vitamins, amino acids, inorganic salts, a buffer, and a fuel, e.g., acetate, succinate, a saccharide/disaccharide/polysaccharide, medium chain fatty acids, and/or optionally nucleotides.
  • a fuel e.g., acetate, succinate, a saccharide/disaccharide/polysaccharide, medium chain fatty acids, and/or optionally nucleotides.
  • Typical components in a growth medium include amino acids (histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine and others); vitamins such as retinol, carotene, thiamine, riboflavin, niacin, biotin, folate, and ascorbic acid; carbohydrate such as glucose, galactose, fructose, or maltose; inorganic salts such as sodium, calcium, iron, potassium, magnesium, zinc; serum; and buffering agents. Additionally, a growth medium may contain phenol red as a pH indication.
  • MIC minimum inhibitory concentration
  • the minimum biofilm eradication concentration (MBEC)” of a compound is defined as the lowest concentration of compound dosed against a previously established bacterial biofilm that leads to a clear well (optical density of less than 0.1) when the treated biofilm is regrown in fresh media, indicating >95% clearance of bacteria.
  • a regrowth assay was used to establish the MBEC of a compound to evaluate the antibiofilm activity.
  • Examples of abbreviations used in the present disclosure include: QAC, quaternary ammonium compound; QPC, quaternary phosphonium compound; AMR, antimicrobial resistance; BAC, benzalkonium chloride; TSC, trivalent sulfonium compound; NMR, nuclear magnetic resonance; HSQC, heteronuclear single quantum coherence spectroscopy; MSSA, methicillin-susceptible Staphylococcus aureus, CA-MRSA, community- acquired methicillin-resistant Staphylococcus aureus; HA-MRSA, hospital-acquired methicillin- resistant Staphylococcus aureus; MIC, minimum inhibitory concentration; DMSO, dimethyl sulfoxide; and PBS, phosphate-buffered saline.
  • QAC quaternary ammonium compound
  • QPC quaternary phosphonium compound
  • AMR antimicrobial resistance
  • BAC benzalkonium chloride
  • TSC trivalent
  • the present disclosure provides a biscationic quaternary phosphonium compound, an antimicrobial composition comprising such a compound, the method of making such a compound, the method of making such an antimicrobial composition, and the methods of using such a compound or composition for antimicrobial use.
  • the biscationic quaternary phosphonium compound is phenyl-free in some embodiments.
  • the compound or the composition provided in the disclosure has an ability to kill, prevent, or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, to attenuate the severity of a microbial infection, or to prevent or inhibit formation of a biofilm or eradicate pre-established biofilms (i.e. antibiofilm use).
  • Scheme 1 shows prior examples of bisQAC and newly synthesized bisQPCs as described in the present disclosure based on the inventors’ research. All compounds have suitable counter ions such as bromide (not shown).
  • Ph P6P-10,10 a structure termed P6P- 10,10.
  • Ph P6P-10,10 a structure termed P6P-10,10 (as shown in Scheme 1).
  • This structure took advantage of the ready availability and stability of phenylsubstituted bisphosphines, which are often employed as ligands for catalysis; presumably, the phenyl substituents stabilize the phosphines and inhibit oxidation and other reactivity.
  • methyl- and ethylsubstituted bisQPC analogs would serve to maximize atom economy within this structural motif and identify minimum structural requirements for strong bioactivity. Further, the methylsubstituted phosphonium compounds would allow for direct comparison of bisQPC to bisQAC activity (Scheme 1, bottom), since the only change in the molecules would be the pnictogen atom (nitrogen or phosphorus) of the cationic head group.
  • the present disclosure provides a phenyl-free biscationic quaternary phosphonium compound, an antimicrobial composition comprising such a compound, and the method of making such an antimicrobial compound or composition, and the method of using such a compound or composition for antimicrobial use.
  • the compound or the composition provided in the disclosure has an ability to kill, prevent, or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, or to attenuate the severity of a microbial infection.
  • the compound or the composition provided in the disclosure has an ability to inhibit or eradicate pre-established biofilms (i.e. antibiofilm use) formed by the microorganisms.
  • the present disclosure provides an antimicrobial composition
  • n is an integer in a range of from 2 to 20
  • L is a linking group being Ci-io alkyl or a cycloalkyl
  • R 1 , R 2, R 3 , or R 4 each is a C 1-12 alkyl or a cycloalkyl, unsubstituted or optional substituted with a functional group selected from the group consisting of -OH, -OR’, -NH 2 , - NHR’, -NR’2, -SH, -SR’, -O-C(O)R’, -C(O)OR’, -C(O)R’, -CF 3 , -OCF 3 , halogen, and any combination thereof;
  • R’ is H or a C 1-4 alkyl
  • X is selected from the group consisting of a halogen, a tosylate, a hydrocarbonate, a carbonate, a sulfate, and an acetate; and m is 1 or 2.
  • X is used for illustration only. It can be monovalent anions or monovalent negative ions such as fluoride ion (F“), chloride ion (Cl-), bromide ion (Br), and iodide (I-). X” may also represent multi-valent anions such as carbonate and sulfate [0048]
  • the present disclosure provides a composition or a product described herein.
  • the antimicrobial composition is a disinfectant configured to be applied to a surface of an object in need thereof.
  • the antimicrobial composition is a pharmaceutical composition configured to be administrated to a subject in need thereof.
  • the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an active ingredient.
  • the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an additive for stabilizing the pharmaceutical composition.
  • n can be in a range of from 6 to 16, from 2 to 12, from 4 to 12, from 6 to 12, from 8 to 12, from 10 to 12, or any other suitable ranges. In some embodiments, n is in a range of from 10 to 12.
  • the linking group L is C1-6 alkyl such as -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, or- CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 -.
  • R 1 , R 2 , R 3 , or R 4 each is a C1-6 alkyl or a cyclohexyl such as cyclohexyl.
  • X is any suitable group for anions.
  • the value of m depends on the valence of the anions. For example, when X a halogen, a tosylate, an acetate, or a hydrocarbonate, m is 2. When X is a carbonate or a sulfate, m is 1. In some embodiments, X is a halogen selected from F, Cl, Br, and I, and m is 2. [0053] In another aspect, the present disclosure provides a compound as described herein, a method of making such a compound, and a method of using such a compound.
  • Examples of a suitable compound include, but are not limited to the compounds from Cl to C21 as shown below:
  • the counter ions may be mX" as described.
  • each of these compounds comprises 2 Br", as counter ions.
  • R PLP-n,n- X the compounds are abbreviated in a format of R PLP-n,n- X.
  • R represents R 1 , R 2 , R 3 , and R 4 , and “Me”, “Ef ’ and “Cy” represent methyl, ethyl and cyclohexyl, respectively.
  • “L” between two P atoms represents the number of the carbon atoms in the linking group or the name of the linking group.
  • X and n are the same as those in the formula (I).
  • Compound (Cl) having the following structure: is coded as M T2P-8,8-Br.
  • the present disclosure provides a method of making the antimicrobial composition as described herein.
  • a method comprises preparing the compound having the formula (I) as described.
  • the compound having the formula [1] can be synthesized as illustrated in Schedule 2.
  • the compound having the formula (I) can be prepared by the alkylation of a suitable bisphosphine starting compound having the formula (II).
  • a suitable electrophile, bearing an alkyl or cycloalkyl backbone as well as a leaving group, can be alkylated under substitution conditions, which can optimize solvent conditions and temperature; two equivalents of the electrophile would be utilized.
  • the method may further comprise mixing an effective amount of the compound having the formula (I) and a carrier.
  • a suitable carrier may include, but are not limited to, a solvent, a carrier, an additive, any other suitable ingredient, or combinations thereof.
  • the present disclosure also provides an antimicrobial composition comprising a compound having the formula (I) as described, and a carrier such as a solvent.
  • the antimicrobial composition can also comprise other ingredients and additives.
  • the content of the compound having the formula (I) can be in any suitable concentration. For example, in some embodiments, such a concentration can be in the range from 0.01 pM to 100 pM, for example, from 0.1 pM to 10 pM.
  • the content of the compound having the formula (I) may be at a concentration of from 0.1 wt% to 5 wt %, for example, in the range of from 0.2 wt.% to 2.5 wt. %.
  • the carrier include but are not limited to a solvent
  • other additives include but are not limited to surfactants, anti-foaming agents, anti-freezing agents, gelling agents, and combinations thereof.
  • the antimicrobial composition may also comprise a pharmaceutically acceptable carrier or excipient
  • a pharmaceutically acceptable carrier or excipient suitable for a solid preparation such as tablets or capsules can be, for example, binders (e.g., acacia, gelatin, dextrin, hydroxypropylcellulose, methylcellulose, polyvinylpyrrolidone), solvents, dispersion media, diluents (e.g., lactose, sucrose, mannitol, com starch, potato starch, calcium phosphate, calcium citrate, crystalline cellulose), lubricants (e.g., magnesium stearate, calcium stearate, stearic acid, talc, anhydrous silicic acid), disintegrants (e.g., com starch, potato starch, carboxymethylcellulose, carboxymethylcellulose calcium, alginic acid), and wetting agents (e.g., sodium laurylsulfate).
  • binders e.g., a
  • a pharmaceutically acceptable carrier or excipient suitable for a liquid preparation can be, for example, aqueous vehicles (e.g., water), suspending agents (e.g., acacia, gelatin, methyl cellulose, carboxymethylcellulose sodium, hydroxymethyl-cellulose, aluminum stearate gel), surfactants (e.g., lecithin, sorbitan monooleate, glycerin monostearate), and non-aqueous vehicles (e.g., glycerin, propylene glycol, vegetable oil).
  • aqueous vehicles e.g., water
  • suspending agents e.g., acacia, gelatin, methyl cellulose, carboxymethylcellulose sodium, hydroxymethyl-cellulose, aluminum stearate gel
  • surfactants e.g., lecithin, sorbitan monooleate, glycerin monostearate
  • non-aqueous vehicles e.g., glycerin, propylene glycol, vegetable oil
  • liquid preparations may contain preservatives (e.g., p-hydroxybenzoic acid methyl ester, p-hydroxybenzoic acid propyl ester), flavors, and/or coloring agents.
  • preservatives e.g., p-hydroxybenzoic acid methyl ester, p-hydroxybenzoic acid propyl ester
  • flavors e.g., p-hydroxybenzoic acid propyl ester
  • coloring agents e.g., p-hydroxybenzoic acid methyl ester, p-hydroxybenzoic acid propyl ester
  • the antimicrobial composition in this disclosure can be formulated to be in any suitable form, including but not limited to liquid, gel and paste.
  • the present disclosure also provides different products as described herein, which comprise the antimicrobial composition.
  • the present disclosure provides a method of killing, preventing, or inhibiting microbial growth.
  • the method comprises applying the antimicrobial composition to a surface of an object in need thereof.
  • a suitable method include but are not limited to pouring, spraying, any other suitable methods and any combinations thereof.
  • such a method comprises administrating the antimicrobial composition to a subject in need thereof.
  • the method of administration may include in vitro or in vivo administration.
  • the types of administration include, are not limited to, topical, oral, parenteral, rectal, intramuscular injection, inhalation, intrathecal, sublingual, nasal, transdermal, and any combination thereof.
  • the antimicrobial composition or the compound is used to kill, prevent, or inhibit growth of at least one group of microorganisms such as bacteria, viruses, yeast, fungi, and protozoa, or to inhibit formation of a biofilm, or disperse or eradicate a pre-established biofilm.
  • this disclosure relates to methods of treating or preventing diseases or conditions comprising administering an effective amount of a composition comprising the quaternary phosphonium compound disclosed herein to a subject in need thereof.
  • the disclosure contemplates that a quaternary phosphonium compound disclosed herein may be used in antimicrobial applications optionally in combination with other antimicrobial agents for prevention of disease onset and treatment.
  • the quaternary phosphonium compound disclosed herein may be used in medical device coatings (medical implants and tools, IV catheters), wound dressings (embedded in gauze bandages), wound rinses (i.e. surgical rinses), wound-vacuum systems, whole body baths (e.g., in combo with bleach baths for treatment of skin flares for atopic dermatitis/eczema), soaps, personal care products (body washes, lotions, soaps) for high-risk patients or for populations with high risk of exposure (e.g. athletes using common sports equipment in gym), and veterinary applications (e.g.
  • this disclosure relates to methods of treating or preventing a microbial infection comprising administering to a subject in need thereof an effective amount of a quaternary phosphonium compound as disclosed herein.
  • the microbial infection is a bacterial, fungal, pest, or viral infection.
  • this disclosure relates to methods of treating or preventing bacterial infections comprising administering or contacting the skin of a subject with formula comprising a quaternary phosphonium compound as disclosed herein to a subject in need thereof.
  • the formula is administered in combination with another antibiotic agent.
  • this disclosure provides a method of using quaternary phosphonium compounds disclosed herein for treating or preventing an Acinetobacter baumannii infection, other bacterial infection, other multidrug resistant bacteria, or other microbial infection by administering an effective amount of quaternary phosphonium compounds disclosed herein to a subject in need thereof.
  • the quaternary phosphonium compound is co-administered with an antibiotic selected from the group comprising of sulfonamides, diaminopyrimidines, quinolones, beta-lactam antibiotics, cephalosporins, tetracyclines, nitrobenzene derivatives, aminoglycosides, macrolide antibiotics, polypeptide antibiotics, nitrofuran derivatives, nitroimidazoles, nicotinic acid derivatives, polyene antibiotics, imidazole derivatives or glycopeptide, cyclic lipopeptides, glycylcyclines and oxazolidinones.
  • an antibiotic selected from the group comprising of sulfonamides, diaminopyrimidines, quinolones, beta-lactam antibiotics, cephalosporins, tetracyclines, nitrobenzene derivatives, aminoglycosides, macrolide antibiotics, polypeptide antibiotics, nitrofur
  • these antibiotics include but are not limited to sulphadiazine, sulphones - [dapsone (DDS) and para-aminosalicylic (PAS)], sulfanilamide, sulfamethizole, sulfamethoxazole, sulphapyridine, trimethoprim, pyrimethamine, nalidixic acids, norfloxacin, ciprofloxacin, cinoxacin, enoxacin, gatifloxacin, gemifloxacin, grepafloxacin, levofloxacin, lomefloxacin, moxifloxacin, ofloxacin, pefloxacin, sparfloxacin, trovafloxacin, penicillins (amoxicillin, ampicillin, azlocillin, carbenicillin, cioxacillin, dicloxacillin, flucioxacillin, hetacillin
  • the subject is diagnosed with a bacterial infection.
  • the subject is diagnosed with bacteremia, pneumonia, staphylococcal food poisoning, necrotizing pneumonia, necrotizing fasciitis, scalded skin syndrome, post-operation bacterial infection, medical device bacterial infection, bacterial infection of the skin, soft tissue bacterial infection, or toxic shock syndrome.
  • this disclosure provides methods of treating or preventing a toxin-mediated bacterial infection comprising administering an effective amount of a quaternary phosphonium compound as disclosed herein to a subject in need thereof, including a subject at risk of, exhibiting symptoms of, or diagnosed with scalded skin syndrome (esp. in neonates), abscesses, necrotizing fasciitis, sepsis, or atopic dermatitis (eczema).
  • scalded skin syndrome esp. in neonates
  • abscesses necrotizing fasciitis
  • sepsis sepsis
  • atopic dermatitis eczema
  • this disclosure provides methods of treating or preventing bacterial infections or acne comprising administering to a subject in need thereof or contacting the skin of a subject in need thereof with a formula comprising of a quaternary phosphonium compound as disclosed herein.
  • the formula is administered in combination with another antibiotic.
  • the subject is at risk of a bacterial infection due to being diagnosed with an abscess, furuncle, cellulitis, folliculitis, atopic dermatitis, psoriasis, impetigo, septic arthritis, brain abscess, bum wound, venous ulcer, diabetic foot ulcer, surgical wound, carbuncle, or meningitis.
  • a flow of wound rinse/irrigation solution can be applied across an open wound surface to achieve wound hydration, to remove deeper debris, and to assist with the visual examination.
  • the disclosure relates to methods of irrigating a wound using a solution comprising a quaternary phosphonium compound as disclosed herein.
  • this disclosure provides methods of using quaternary phosphonium compounds disclosed herein for killing microbes, preventing or inhibiting microbe growth, preventing a biofilm formation, or preventing the spread of an Acinetobacter baumannii infection, other bacterial infection, other multidrug resistant bacteria, or other microbial infection by sanitizing a surface, e.g., by contacting the surface with a solid (e.g., powder), liquid, or spray composition, with a quaternary phosphonium compound disclosed herein in an effective amount.
  • this disclosure provides methods of preventing cellular infections comprising applying a quaternary phosphonium compound disclosed herein on top of or inside a cell growth medium.
  • this disclosure provides methods of preventing plant microbial infections comprising applying a quaternary phosphonium compound disclosed herein to the exterior, leaf, seed, or stem of a plant. In some embodiments, this disclosure provides methods of preventing plant microbial infections comprising applying a quaternary phosphonium compound disclosed herein on top of or into soil, dirt, sand, or other medium from which roots of the plant reside.
  • compositions and devices comprising a quaternary phosphonium compound disclosed herein.
  • this disclosure provides soaps and disinfectant products comprising a quaternary phosphonium compound disclosed herein.
  • Contemplated topical formulations for skin flares i.e., for atopic dermatitis or other infections related to a disrupted skin barrier
  • another drug such as a topical steroid, anti-inflammatory agent, and promoter of skin barrier function or skin moisturizer.
  • this disclosure provides a container configured to create a liquid spray comprising a quaternary phosphonium compound disclosed herein.
  • this disclosure provides a pharmaceutical formulation comprising a quaternary phosphonium compound disclosed herein and a pharmaceutically acceptable excipient.
  • the pharmaceutical formulation is in the form of a lotion, liquid, or gel.
  • the pharmaceutical formulation is in the form of a particle, bead, tablet, capsule, pill, or injectable solution.
  • the injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as water, dimethyl sulfoxide, mannitol, 1,3-butanediol, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.
  • suitable non-toxic, parenterally-acceptable diluents or solvents such as water, dimethyl sulfoxide, mannitol, 1,3-butanediol, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.
  • the pharmaceutical formulation can also include any type of pharmaceutically acceptable excipients, additives, or vehicles.
  • diluents or fillers such as dextrates, dicalcium phosphate, calcium sulphate, lactose, cellulose, kaolin, mannitol, sodium chloride, dry starch, sorbitol, sucrose, inositol, powdered sugar, bentonite, microcrystalline cellulose, or hydroxypropyl methylcellulose, may be added to the composition to increase the bulk of the composition.
  • the formulation is a directly compressible composition comprising a quaternary phosphonium compound disclosed herein but no excipients, additives, or vehicles.
  • the disclosure provides a pharmaceutical or cosmetic formulation comprising a quaternary phosphonium compound disclosed herein and a pharmaceutically acceptable excipient or cosmetically acceptable excipient.
  • the disclosure provides a liquid or gel formulation optionally further comprising an antibacterial agent, a topical steroid, an anti-inflammatory agent, a promoter of skin barrier function, a skin moisturizer, or combinations thereof.
  • the antibacterial agent is daptomycin, linezolid, vancomycin, nafcillin, cefazolin, dicloxacillin, clindamycin, rifampin, or sulfamethoxazole-trimethoprim (Bactrim).
  • the disclosure provides a wound dressings or wound rinse comprising a quaternary phosphonium compound disclosed herein wherein the wound dressing comprises an absorbent pad and optionally an adhesive.
  • the disclosure provides disinfectant sprays or wipes formulation for surfaces and fomites comprising a quaternary phosphonium compound disclosed herein.
  • this disclosure provides a medical device coated with a quaternary phosphonium compound as disclosed herein.
  • the medical device is a screw, pin, plate, rod, disk, needle, catheter, tube, stent, pacemaker, defibrillators (ICDs), artificial hip or knee joint/implant, breast implant, intra-uterine device, ear tube, contact lens, or implantable pump.
  • ICDs defibrillators
  • this disclosure provides a surgical tool coated with a quaternary phosphonium compound as disclosed herein.
  • the surgical tool is a forceps, tweezers, scalpel, knife, scissors, retractor, needle, gauze, sponge, suction, staple, stapler, clip, laparoscopic instrument, electrosurgical cauterizer, ultrasonic device, camera, camera lens, fiber optic cable, insufflator, needle, bronchoscope, cystoscope, saw, or robotic arm.
  • the disclosure provides a wound dressing comprising a quaternary phosphonium compound as disclosed herein wherein the wound dress comprises an absorbent pad and optionally an adhesive optionally in combination with another antibiotic agent.
  • the wound dressing is a foam or compression dressing or a cover dressing such as wraps, gauze and tape.
  • the wound dressing comprises alginate or collagen.
  • the wound dressing is a hydrocolloid dressing, e.g., carboxy-methylcellulose and gelatin optionally in a polyurethane foam or film, optionally comprising one or more agents selected from pectin, a polysaccharide, and an adhesive.
  • the wound dressing is a hydrogel.
  • Hydrogels are polymers that contain a high content of hydroxy and/or carboxyl containing monomers or salts thereof, e.g., vinyl alcohol, acrylic acid, 2-hydroxyethylmethacrylate, ethylene glycol dimethacrylate monomers, which are co-polymers to provide varying degrees of hydration. Due to the hydrophilic monomers, the hydrogels typically absorb water.
  • Contemplated hydrogel dressings include: amorphous hydrogel, which are a free-flowing gel that are typically distributed in tubes, foil packets and spray bottles; an impregnated hydrogel, which are typically saturated onto a gauze pad, nonwoven sponge ropes and/or strips; or a sheet hydrogel which are gel held together by a fiber mesh.
  • the disclosure provides a wound rinse comprising a quaternary phosphonium compound as disclosed herein optionally containing normal saline, sterile water, a detergent, a surfactant, a preservative, or iodine.
  • the disclosure provides a kit comprising a container comprising a quaternary phosphonium compound as disclosed herein optionally comprising a second container comprising a rinse solution or containing surgical device or tool, normal saline, sterile water, a detergent, a surfactant, a preservative, iodine, hydrogen peroxide, or sodium hypochlorite or other compound disclosed herein.
  • the disclosure provides a cosmetic formulation comprising a quaternary phosphonium compound as disclosed herein and cosmetically acceptable excipient or additive.
  • the disclosure relates to a solid or liquid soap or lotion comprising a quaternary phosphonium compound as disclosed herein and a fatty acid.
  • additives can be selected from the group consisting of oily bodies, surfactants, emulsifiers, fats, waxes, pearlescent waxes, bodying agents, thickeners, superfatting agents, stabilizers, polymers, silicone compounds, lecithins, phospholipids, biogenic active ingredients, deodorants, antimicrobial agents, antiperspirants, film formers, antidandruff agents, swelling agents, insect repellents, hydrotropes, solubilizers, preservatives, perfume oils and dyes.
  • additives are selected from the group consisting of surfactants, emulsifiers, fats, waxes, stabilizers, deodorants, antiperspirants, antidandruff agents, and perfume oils.
  • this disclosure provides a cell growth medium comprising a quaternary phosphonium compound disclosed herein.
  • Amphiphilic QAC and QPC compounds containing alkyl chain(s) of 10 to 12 carbon atoms display optimal efficacy as disinfectants. Therefore, a series of compounds with alkyl chain lengths between 8 and 16 carbons were prepared to bracket this hypothesized ideal nonpolar region.
  • the reaction was stirred for 5 hours at 75 °C, and then allowed to further stir at room temperature for 18 h.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting colorless, viscous oil was triturated with diethyl ether to form a white solid.
  • 1,2-Bis(dimethylphosphino)ethane (0.40 mL, 0.36 g, 2.4 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (0.8 mL) and 1 -bromononane (1.30 mL, 1.41 g, 6.8 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 7 hours at 72 °C, and then cooled to room temperature.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting waxy, white solid was triturated with 8 mL of hexanes and the slurry was placed into a -20 °C freezer overnight.
  • the product was isolated via vacuum filtration, then triturated again with 12 mL of diethyl ether and vacuum filtered to afford a white waxy solid.
  • 1,2-Bis(dimethylphosphino)ethane (0.40 mL, 0.36 g, 2.4 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (0.7 mL) and 1 -bromodecane (1.30 mL, 1.39 g, 6.3 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 75 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 5 hours at 75 °C, and then allowed to further stir at room temperature for 18 h.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting colorless, viscous oil was triturated with diethyl ether to form a white solid.
  • 1,2-Bis(dimethylphosphino)ethane (0.40 mL, 0.36 g, 2.4 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (0.6 mL) and 1 -bromoundecane (1.60 mL, 1.69 g, 7.2 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 5 hours at 72 °C, and then allowed to further stir at room temperature overnight.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days.
  • 1,2-Bis(dimethylphosphino)ethane (0.30 mL, 0.27 g, 1.8 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (0.6 mL) and 1 -bromododecane (1.30 mL, 1.35 g, 5.4 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 5 hours at 72 °C, and then allowed to further stir at room temperature overnight.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days.
  • the reaction was stirred for 7 hours at 72 °C, and then cooled to room temperature.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting waxy, white solid was triturated with 8 mL of hexanes and the slurry was placed into a -20 °C freezer overnight.
  • the product was isolated via vacuum filtration, then triturated again with 12 mL of diethyl ether and vacuum filtered to afford a fine white powder.
  • 1,2-Bis(dimethylphosphino)ethane (0.30 mL, 0.27 g, 1.8 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (0.7 mL) and 1 -bromohexadecane (1.65 mL, 1.65 g, 5.4 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • 1,2-Bis(diethylphosphino)ethane (0.25 mL, 0.22 g, 1.1 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (1 mL) and 1 -bromooctane (0.55 mL, 0.61 g, 3.2 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 5 h at 70 °C, then cooled to room temperature and stirred for 15 h. At the end of the stir time, the volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 1 day.
  • the reaction was stirred for 6 h at 75 °C, then cooled to room temperature.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days.
  • 1,2-Bis(diethylphosphino)ethane (0.28 mL, 0.25 g, 1.2 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (1 mL) and 1 -bromodecane (0.75 mL, 0.80 g, 3.6 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 6 h at 75 °C, then cooled to room temperature.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days.
  • 1,2-Bis(diethylphosphino)ethane (0.28 mL, 0.25 g, 1.2 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (1 mL) and 1 -bromododecane (0.87 mL, 0.90 g, 3.6 mmol) were added to the vial, also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 75 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 6 h at 75 °C, then cooled to room temperature.
  • the volatiles were removed from the vial using rotary evaporation.
  • the resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days.
  • 1,2-Bis(diethylphosphino)ethane (0.26 mL, 0.23 g, 1.1 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (1 mL) and 1 -bromotetradecane (1.0 mL, 0.93 g, 3.4 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 5 h at 70 °C, then cooled to room temperature and stirred for 15 h. At the end of the stir time, the volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 1 day.
  • 1,2-Bis(diethylphosphino)ethane (0.26 mL, 0.23 g, 1.1 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition.
  • Acetonitrile (1 mL) and 1 -bromohexadecane (1.0 mL, 1.0 g, 3.3 mmol) were added to the vial also by syringe.
  • the reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial.
  • the reaction was stirred for 5 h at 70 °C, then cooled to room temperature and stirred for 15 h. At the end of the stir time, the volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ⁇ 2 mL of dichloromethane, layered with ⁇ 30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 1 day.
  • 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromooctane (0.18 mL, 0.20 g, 1.0 mmol) were added to the reaction vial via needle and syringe.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation.
  • the resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer.
  • 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromononane (0.20 mL, 0.22 g, 1.0 mmol) were added to the reaction vial via needle and syringe.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by loosening the cap twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation.
  • the resulting white powder was purified via trituration with diethyl ether (approx. 30 mL) and isolated by vacuum filtration, resulting in the product as a white solid.
  • 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromodecane (0.22 mL, 0.23 g, 1.0 mmol) were added to the reaction vial via needle and syringe.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation.
  • the resulting white gel was dissolved in (approx. 2 mL) chloroform-d layered with (approx. 30 mL) hexanes and place in a -20 °C freezer.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C vented by puncturing the cap with a needle twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation.
  • the resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer.
  • 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromododecane (0.25 mL, 0.26 g, 1.0 mmol) were added to the reaction vial via needle and syringe.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation.
  • the resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer.
  • 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromotridecane (0.27 mL, 0.28 g, 1.0 mmol) were added to the reaction vial via needle and syringe.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time.
  • the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was washed with diethyl ether. Residual solvent was again removed by rotary evaporation, resulting in the product as a white, glassy solid.
  • 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromotetradecane (0.31 mL, 0.29 g, 1.0 mmol) were added to the reaction vial via needle and syringe.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation.
  • the resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer.
  • the reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup.
  • the reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation.
  • the resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer.
  • the proton and carbon NMR signals can be corrected with each other in one plot to identify the spectroscopically unique methyl groups in the molecule and all methylene signals.
  • the counterions for the compounds provided in the present disclosure is not limited to bromide only.
  • the counterions may include, but are not limited to, F", Cl", Br", I", tosylate, carbonate, sulfate, and any combination thereof.
  • an exemplary compound with counterions different from Br” may be synthesized using the schemes described with Br- counterions.
  • Br" counterions in an exemplary compound may be replaced by other counter ions after the synthesis of such a compound with Br- counterions.
  • MIC Minimum Inhibitory Concentration
  • RBC lysis assays were performed on mechanically defibrinated sheep blood (Hemostat Labs: DSB030). An aliquot of 1.5 mL blood was placed into a microcentrifuge tube and centrifuged at 3,800 rpm for ten minutes. The supernatant was removed, and the cells were resuspended with 1 mL of phosphate-buffered saline (PBS). The suspension was centrifuged as described above, the supernatant was removed, and cells were resuspended 4 additional times in 1 mL PBS. The final cell suspension was diluted twentyfold with PBS.
  • PBS phosphate-buffered saline
  • the absorbance of the supernatant was measured with a UV spectrometer at a 540 nm wavelength.
  • the concentration inducing 20% RBC lysis was then calculated for each compound based upon the absorbances of the TritonX and PBS controls.
  • Aqueous DMSO controls were conducted as appropriate for each compound.
  • the inventors therefore, were interested if the identity of the cationic species in these amphiphiles would change dynamic surface tension as a physical property that could be correlated to observed bioactivity data.
  • the dynamic surface tension experiments were completed at three difference concentrations (100, 300 and 1000 ppm) to elucidate any concentration dependance upon the data.
  • FIG. 1 shows characterization data for bisQAC and bisQPC compounds 12(2)12 and MeP2P-12,12, including dynamic surface tension (mN/m) data when plotted as a function of surface age (ms) at concentrations of 100, 300 and 1000 ppm. Bromide counter ions omitted for clarity.
  • These bacteria include the Gram-positive strains of methicillin-susceptible Staphylococcus aureus [MSSA; SHI 000], community-acquired methicillin-resistant Staphylococcus, aureus [CA-MRSA; USA 300-0114], hospital-acquired methicillin-resistant Staphylococcus aureus [HA-MRSA; ATCC 33591] and the Gram-negative strains of Enterococcus faecalis [OG1RF], Escherichia coli [MC4100], Acinetobacter baumannii [ATCC 17498] and Pseudomonas aeruginosa [PAO1].
  • MSSA methicillin-susceptible Staphylococcus aureus
  • CA-MRSA community-acquired methicillin-resistant Staphylococcus, aureus
  • H-MRSA hospital-acquired methicillin-resistant Staphylococcus aureus
  • PAO1 Pseudom
  • Table 1 summarizes biological activity and lysis20 data of alkyl bisQPC compounds.
  • Biofilm eradication experiments were performed using a pegged-lid microtiter plate assay to determine the MBEC values for compounds of interest, as previously described. See Yang, H; Abouelhassan, ⁇ .; Burch, G.M.; Kallifidas, D.; Huang, G; Yousaf, H; Jin,S.; Luesch, H.; Huigens, RW. A Highly Potent Class of Halogenated Phenazine Antibacterial and Biofilm-Eradicating Agents Accessed Through a Modular Wohl-Aue Synthesis. Sei. Rep. 2017, 7, 2003; andRaval, Y. S., Flurin, L., Mohamed, A., Greenwood-Quaintance, K.
  • the pegged lid was then removed, washed with PBS, and transferred to another 96-well plate containing 2-fold serial dilutions of the test compounds (the “challenge plate”).
  • the total volume in each well was 150 mL, comprising of 75 mL of compound diluted in water/DMSO, with a starting DMSO concentration of 2.5 %, and 75 mL of TSB. Plates were incubated statically at 37 °C for 24 hours. Next, the pegged lids were transferred to a fresh 96-well plate containing 180 mL of TSB and incubated overnight at 37 °C.
  • MBEC values were determined as the lowest test concentration that resulted eradicated biofilm (i.e., wells displaying no turbidity in the final plate).
  • the inventors sought to further investigate the antibacterial properties while comparing the effects of the heteroatom on activity. To do so, the inventors determined the minimum biofilm eradication concentration (MBEC) of bisQPC M T2P- 12, 12 and analogous bisQAC 12(2)12 using a pegged-lid microtiter plate assay. Both quaternary amphiphiles demonstrated an ability to eradicate established biofilm against three of the strains tested (Table 2).
  • MBEC biofilm eradication concentration
  • Table 2 shows minimum Biofilm Eradication Concentrations of bisQPC Me P2P- 12,12 and bisQAC 12(2)12, as well as QAC standard DDAC.
  • MBEC values for bisQAC 12(2)12 differed from a previous report, which highlights the effect that experimental method and growth conditions may have in antibacterial evaluations. While only minimal difference in potency was observed between the ammonium and phosphonium analogs, these results demonstrated that the novel cationic biocides provided in the present disclosure have superior biofilm eradication potency over alkyl commercially available QACs like didecyldimethylammonium chloride (DDAC).
  • DDAC didecyldimethylammonium chloride
  • a novel series of alkyl based bisQPC compounds has been synthesized and evaluated for the ability to serve as disinfectants against a panel of bacteria, including those with previously reported AMR.
  • the resulting compounds continue to demonstrate the effectiveness of bisQPCs as viable antibacterials, with improved bioactivity against multiple antibiotic-resistant bacterial strains over a commercial benchmark of BAC.
  • the substituting groups of methyl, ethyl, cyclohexyl and phenyl appended to the phosphonium cations in the bisQPCs imparted only subtle differences in bioactivity across this R P2P-n,n structural motif.
  • the experimental compounds shown above include two carbons in the linking group.
  • the linking group is not limited to two carbons only.
  • the compounds with longer linking groups for example, with 3-6 carbons or a cyclic group such as a cyclohexyl group are expected to have excellent antimicrobial performance. Examples of these compounds include, but are not limited to the following:
  • Each of these compounds comprises m X", for example, 2 Br", as counter ions.
  • the compounds C22, C23, and C24 are coded as M T4P-10,10- Br, M T6P-10,10-Br, M TCyP-10,10-Br.
  • the compounds C22, C23, and C24 are shown for illustration pinpose only.
  • the methyl group can be replaced with other alkyl groups such as ethyl and cyclohexyl.
  • the linking group L may have three or five carbons.
  • the value of n can be in a range of from 6 to 12, for example.
  • the counterions can be any other type as described herein.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Dentistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

A compound having formula (I) and an antimicrobial composition comprising a compound having the formula (I) are provided: (I). In the compound having the formula (I), n is an integer in a range of from 2 to 20, ad A is a linking group being C1-10 alkyl or a cycloalkyl. R1, R2, R3, or R4 each is a C1-12 alkyl or a cycloalkyl, unsubstituted or optional substituted with a functional group selected from the group consisting of -OH, -OR', -NH2, -NHR', -NR'2, -SH, -SR', -O-C(O)R', -C(O)OR', -C(O)R', -CF3, -OCF3, halogen, and any combination thereof. R' is H or a C1-4 alkyl. X is selected from the group consisting of a halogen, a tosylate, a hydrocarbonate, a carbonate, a sulfate, and an acetate, and m is 1 or 2. The methods of making and using the compound and the composition are also provided.

Description

PHENYL-FREE BISCATIONIC QUATERNARY PHOSPHONIUM COMPOUNDS AS ANTIMICROBIAL AGENTS
PRIORITY CLAIM AND CROSS-REFERENCE
[0001] This application claims the priority benefit of U. S. Provisional Application No. 63/425,604, filed November 15, 2022, and U.S. Provisional Application No. 63/431,801, filed December 12, 2022, which applications are expressly incorporated by reference herein in their entirety.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR
DEVELOPMENT
[0002] This invention was made with government support under R35 GM119426 awarded by the National Institute of General Medical Sciences, and under CHE-1827930 and CHE-2018399. The government has certain rights in the invention.
FIELD OF THE INVENTION
[0003] The disclosure relates to antimicrobial compositions and related methods generally. More particularly, the disclosed subject matter relates to a biscationic quaternary phosphonium compound, a composition comprising the biscationic quaternary phosphonium compound, the methods of making the same, and the methods of using the same as antimicrobial agents.
BACKGROUND
[0004] For approximately 100 years, quaternary ammonium compounds (QACs) have been employed as successful tools to mitigate the transfer of pathogenic bacteria. Over much of this time, society has heavily relied on just a handful of these disinfecting amphiphilic compounds, with the most visible commercial disinfectant of this class being benzalkonium chloride (BAC), a mixture of long-chained alkyl dimethylbenzylammonium chlorides. While mainstay disinfectants such as BAC have proven highly valuable for decades, they are now showing vulnerabilities as bacteria continue to evolve antimicrobial resistance mechanisms.
Notable pathogens such as A. baumarmii and P. aeruginosa have developed sufficient resistance mechanisms towards common disinfectants that they are renewing the problem of bacterial infections in hospital, domestic and commercial settings.
SUMMARY
[0005] This Summary is provided to briefly indicate the nature and substance of the invention. It is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims.
[0006] The present disclosure provides a biscationic quaternary phosphonium compound, an antimicrobial composition comprising such a compound, the method of making such a compound, the method of making such an antimicrobial composition, and the methods of using such a compound or composition for antimicrobial use. The biscationic quaternary phosphonium compound is phenyl-free in some embodiments. The compound or the composition provided in the disclosure has an ability to kill or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, to attenuate the severity of a microbial infection, or to prevent or inhibit formation of a biofilm or eradicate pre-established biofilms (i.e. antibiofilm use).
[0007] In accordance with some embodiments, the present disclosure provides an antimicrobial composition comprising a compound having the formula
Figure imgf000004_0001
wherein: n is an integer in a range of from 2 to 20,
L is a linking group being C1-10 alkyl or a cycloalkyl, R1, R2, R3, or R4 each is a C1-12 alkyl or a cycloalkyl, unsubstituted or optional substituted with a functional group selected from the group consisting of -OH, -OR’, -NH2, - NHR’, -NR’2, -SH, -SR’, -O-C(O)R’, -C(O)OR’, -C(O)R’, -CF3, -OCF3, halogen, and any combination thereof;
R’ is H or a C1-4 alkyl; X is selected from the group consisting of a halogen, a tosylate, a hydrocarbonate, a carbonate, a sulfate, and an acetate; and m is 1 or 2.
[0008] The present disclosure provides a composition or a product described herein. In some embodiments, the antimicrobial composition is a disinfectant configured to be applied to a surface of an object in need thereof. In some embodiments, the antimicrobial composition is a pharmaceutical composition configured to be administrated to a subject in need thereof. The pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an active ingredient. In some embodiments, the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an additive for stabilizing the pharmaceutical composition.
[0009] In another aspect, the present disclosure provides a compound as described herein, a method of making such a compound, and a method of using such a compound. [0010] In another aspect, the present disclosure provides a method of making the antimicrobial composition as described herein. In some embodiments, such a method comprises preparing the compound having the formula (I) as described. The method may further comprise mixing an effective amount of the compound having the formula (I) and a carrier.
[0011] In another aspect, the present disclosure also provides different products as described herein, which comprise the antimicrobial composition.
[0012] In another aspect, the present disclosure provides a method of killing, preventing, or inhibiting microbial growth. For example, in some embodiments, the method comprises applying the antimicrobial composition to a surface of an object in need thereof. In some embodiments, such a method comprises administrating the antimicrobial composition to a subject in need thereof. The antimicrobial composition or the compound is used to kill, prevent, or inhibit growth of at least one group of microorganisms such as bacteria, viruses, yeast, fungi, and protozoa, or to inhibit formation of a biofilm, or disperse or eradicate a pre-established biofilm. BRIEF DESCRIPTION OF THE DRAWINGS
[0013] The present disclosure is best understood from the following detailed description when read in conjunction with the accompanying drawings. It is emphasized that, according to common practice, the various features of the drawings are not necessarily to scale. On the contrary, the dimensions of the various features are arbitrarily expanded or reduced for clarity. Like reference numerals denote like features throughout specification and drawings.
[0014] FIG. 1 shows characterization data for structurally analogous bisQAC and bisQPC compounds 12(2)12 and MeP2P-12,12 with bromide counter ions. Shown are dynamic surface tension (mN/rn) data when plotted as a function of surface age (ms) at three concentrations of 100, 300 and 1000 ppm.
DETAILED DESCRIPTION
[0015] For purposes of the description hereinafter, it is to be understood that the embodiments described below may assume alternative variations and embodiments. It is also to be understood that the specific articles, compositions, and/or processes described herein are exemplary and should not be considered as limiting.
[0016] In the present disclosure, the singular forms “a,” “an,” and “the” include the plural reference, and reference to a particular numerical value includes at least that particular value, unless the context clearly indicates otherwise. Thus, for example, a reference to “a compound” or “a structure” is a reference to one or more of such compounds or structures and equivalents thereof known to those skilled in the art, and so forth.
[0017] When values are expressed as approximations, by use of the antecedent “about” or “approximately,” it will be understood that the particular value forms another embodiment. As used herein, “about X” (where X is a numerical value) preferably refers to ±10% of the recited value, inclusive. For example, the phrase “about 8” preferably refers to a value of 7.2 to 8.8, inclusive; as another example, the phrase “about 8%” preferably (but not always) refers to a value of 7.2% to 8.8%, inclusive.
[0018] Where present, all ranges are inclusive and combinable. For example, when a range of “1 to 5” is recited, the recited range should be construed as including ranges “1 to 4”, “1 to 3”, “1-2”, “1-2 & 4-5”, “1-3 & 5”, “2-5”, and the like. In addition, when a list of alternatives is positively provided, such listing can be interpreted to mean that any of the alternatives may be excluded, e.g., by a negative limitation in the claims. For example, when a range of “1 to 5” is recited, the recited range may be construed as including situations whereby any of 1, 2, 3, 4, or 5 are negatively excluded; thus, a recitation of “1 to 5" may be construed as “1 and 3-5, but not 2”, or simply “wherein 2 is not included.” It is intended that any component, element, attribute, or step that is positively recited herein may be explicitly excluded in the claims, whether such components, elements, attributes, or steps are listed as alternatives or whether they are recited in isolation.
[0019] The term “antimicrobial” refers to an ability to kill, prevent, or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, or to attenuate the severity of a microbial infection. The antimicrobial compounds or compositions of the present invention are compounds or compositions that may be used for cleaning or sterilization, or may be used in the treatment of disease and infection. The applications may include both in vitro and in vivo antimicrobial uses. “Applying” an antimicrobial composition may include administrating a composition into a human or animal subject.
[0020] The term “biofilm” as used herein refer to a film formed by a group of microorganisms adhered together. The term “antibiofilm” as used herein refer to an ability to kill, disperse and/or eradicate a pre-established biofilm.
[0021] The term “alkyl” as used herein refers to a straight chain, cyclic, branched or unbranched saturated or unsaturated hydrocarbon chain containing 1-25 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n- septyl, n-octyl, n-nonyl, cyclo-hexyl, and the like. The alkyl may be straight or branched, and can be optionally substituted. “A C1-12 alkyl” as used herein refers to an alkyl group having a number of carbon atoms selected from 1 to 12. Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (referred to as an “alkenyl” or “alkynyl,” respectively). Representative straight chain and branched alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl, 1 -pentenyl, 2-pentenyl, 3 -methyl- 1-butenyl, 2-methyl-2- butenyl, 2,3- dimethyl-2-butenyl, and the like; while representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1 -pentynyl, 2-pentynyl, 3- methyl- 1-butynyl, and the like.
[0022] The term “optionally substituted” means that group in question may be unsubstituted or it may be substituted one or several times, such as 1 to 3 times or 1 to 5 times. For example, an alkyl group that is “optionally substituted” with 1 to 5 chloro atoms, may be unsubstituted, or it may contain 1, 2, 3, 4, or 5 chlorine atoms. Substituted chemical moieties include one or more substituents that replace hydrogen.
[0023] The term “microorganism” or “microbe” as used herein refers to a small (often, but not always, microscopic) organism that is typically, but not exclusively, single cellular, and includes organisms from the kingdoms bacteria, archaea, protozoa, and fungi. The present disclosure is primarily directed to microorganisms that are pathogenic and capable of causing disease. Such microbes may be detected from a sample. In embodiments, microorganism includes bacteria and fungi capable of causing disease, particularly disease in humans and other mammals and animals in need of treatment.
[0024] The term “object” used herein refers to any article having a solid surface. An object may also include a body part of a subject. A composition to be applied to a surface of an object may be in a form of liquid or aerosol.
[0025] A “subject” refers any animal, preferably a human being such as a patient, livestock, rodent, monkey, or domestic pet. In some embodiments, the subject may be exhibiting symptoms of, at risk of, or diagnosed with a disease or condition by analysis of a sample. The term “sample” can refer to a tissue sample, cell sample, a fluid sample, and the like. A sample may be taken from a host subject The tissue sample can include hair, buccal swabs, blood, saliva, semen, muscle, or tissue from any internal organ. The fluid may be, but is not limited to, urine, blood, ascites, pleural fluid, spinal fluid, semen, wound exudates, sputum, fecal matter, saliva, and the like. The body tissue can include, but is not limited to, skin, muscle, endometrial, uterine, and cervical tissue. While a sample, in the context of the present disclosure, is primarily a biological sample (e.g., from a living host) the sample may also be an environmental sample suspected of contamination by microbes, such as a water sample, food sample, soil sample, and the like. Although a liquid sample and some solid samples may be used as a test sample without modification for testing directly, if a solid sample is to be made into liquid form for testing and/or a liquid sample is to be diluted, a test sample may be made by reconstituting, dissolving, or diluting the sample in a fluid such as water, buffered saline, and the like.
[0026] As used herein, the terms “administration" or “administrating” is understood to encompass any type of application to a subject including in vitro or in vivo administrations. For example, the types of administration include, are not limited to, topical, oral, parenteral, rectal, intramuscular injection, inhalation, intrathecal, sublingual, nasal, transdermal, and any combination thereof. The composition to be administrated may be in a form of a solid dose form such as a tablet or a capsule, a liquid form, or an aerosol.
[0027] As used herein, the terms “prevent" and “preventing” include the prevention of the recurrence, spread or onset. It is not intended that the present disclosure be limited to complete prevention. In some embodiments, the onset is delayed, or the severity is reduced. [0028] As used herein, the terms “treat” and “treating” are not limited to the case where the subject (e.g. patient) is cured and the disease is eradicated. Rather, embodiments of the present disclosure also contemplate treatment that merely reduces symptoms, and/or delays disease progression.
[0029] The term “effective amount” or “therapeutically effective amount" refers to that amount of a compound or pharmaceutical composition described herein that is sufficient to effect the intended application including, but not limited to, disease treatment, as illustrated below.
The therapeutically effective amount can vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. The term also applies to a dose that will induce a particular response in target cells, e.g., reduction of platelet adhesion and/or cell migration. The specific dose will vary depending on, for example, the particular compounds chosen, the dosing regimen to be followed, whether it is administered in combination with other agents, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.
[0030] A “linking group” refers to any variety of molecular arrangements that can be used to bridge molecular moieties together.
[0031] In some embodiments, this disclosure contemplates derivatives of compounds disclosed herein. As used herein, the term “derivative” refers to a structurally similar compound that retains sufficient functional attributes of the identified analogue. The derivative may be structurally similar because it is lacking one or more atoms, substituted, a salt, or alternative salt, an ester, in different hydration/oxidation states, or because one or more atoms within the molecule are switched, such as, but not limited to, replacing an oxygen atom with a sulfur atom, or replacing an amino group with a hydroxy group. [0032] The term “substituted” refers to a molecule wherein at least one hydrogen atom is replaced with a substituent When substituted, one or more of the groups are “substituents.” The molecule may be multiply substituted. In the case of an oxo substituent (“=O”), two hydrogen atoms are replaced. Example substituents may include halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, amine, and ester.
[0033] The terms, “cell culture” or “growth medium” or “media” refers to a composition that contains components that facilitate cell maintenance and growth through protein biosynthesis, such as vitamins, amino acids, inorganic salts, a buffer, and a fuel, e.g., acetate, succinate, a saccharide/disaccharide/polysaccharide, medium chain fatty acids, and/or optionally nucleotides. Typical components in a growth medium include amino acids (histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine and others); vitamins such as retinol, carotene, thiamine, riboflavin, niacin, biotin, folate, and ascorbic acid; carbohydrate such as glucose, galactose, fructose, or maltose; inorganic salts such as sodium, calcium, iron, potassium, magnesium, zinc; serum; and buffering agents. Additionally, a growth medium may contain phenol red as a pH indication.
[0034] The term “minimum inhibitory concentration (MIC)” means the lowest concentration of an antimicrobial agent that will inhibit the visible growth of a microorganism after overnight incubation. MIC values against bacteria, for example, the Gram-positive Staphylococcus aureus and Enterococcus faecalis and the Gram-negative Escherichia coli and Pseudomonas aeruginosa were determined by standard methods. See also P. A. Wayne, Methods for Dilution Antimicrobial Tests for Bacteria that Grow Aerobically; Approved Standard, Ninth Edition, 2012, CLSI Document M07-A9, Vol. 32 No. 2.
[0035] The term “the minimum biofilm eradication concentration (MBEC)” of a compound is defined as the lowest concentration of compound dosed against a previously established bacterial biofilm that leads to a clear well (optical density of less than 0.1) when the treated biofilm is regrown in fresh media, indicating >95% clearance of bacteria. A regrowth assay was used to establish the MBEC of a compound to evaluate the antibiofilm activity.
[0036] Examples of abbreviations used in the present disclosure include: QAC, quaternary ammonium compound; QPC, quaternary phosphonium compound; AMR, antimicrobial resistance; BAC, benzalkonium chloride; TSC, trivalent sulfonium compound; NMR, nuclear magnetic resonance; HSQC, heteronuclear single quantum coherence spectroscopy; MSSA, methicillin-susceptible Staphylococcus aureus, CA-MRSA, community- acquired methicillin-resistant Staphylococcus aureus; HA-MRSA, hospital-acquired methicillin- resistant Staphylococcus aureus; MIC, minimum inhibitory concentration; DMSO, dimethyl sulfoxide; and PBS, phosphate-buffered saline.
[0037] The present disclosure provides a biscationic quaternary phosphonium compound, an antimicrobial composition comprising such a compound, the method of making such a compound, the method of making such an antimicrobial composition, and the methods of using such a compound or composition for antimicrobial use. The biscationic quaternary phosphonium compound is phenyl-free in some embodiments. The compound or the composition provided in the disclosure has an ability to kill, prevent, or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, to attenuate the severity of a microbial infection, or to prevent or inhibit formation of a biofilm or eradicate pre-established biofilms (i.e. antibiofilm use).
[0038] In order to combat the growing problem of antimicrobial resistance (AMR), ongoing efforts toward the synthesis and antimicrobial evaluation of novel QACs are underway. Along these lines, the inventors have systematically evaluated a variety of QAC structural motifs to identify molecular features that can inform the rational design of new classes of disinfectants. The inventors have recently advanced to machine learning techniques to predict bioactivity of proposed compounds. Despite the development of over 700 different compounds by the inventors in the past decade, concerns remain about the cross-resistance in AMR mechanisms between established QACs such as BAC into any novel QAC.
[0039] Accordingly, exploration of molecules with markedly different cationic amphiphilic heads, such as quaternary phosphonium compounds (QPCs) and trivalent sulfonium compounds (TSCs), are being pursued.
[0040] Scheme 1 shows prior examples of bisQAC and newly synthesized bisQPCs as described in the present disclosure based on the inventors’ research. All compounds have suitable counter ions such as bromide (not shown).
[0041] Scheme 1.
Figure imgf000012_0001
[0042] The inventors’ investigation into alternative amphiphiles resulted in the discovery of highly effective novel biscationic QPC compounds, highlighted by a structure termed P6P- 10,10. For clarity within this present disclosure, it is labeled hereafter as PhP6P-10,10 (as shown in Scheme 1). This structure took advantage of the ready availability and stability of phenylsubstituted bisphosphines, which are often employed as ligands for catalysis; presumably, the phenyl substituents stabilize the phosphines and inhibit oxidation and other reactivity. The high degree of antimicrobial efficacy observed with PhP6P-10,10 towards both Gram-positive and Gram-negative pathogens, including highly resistant pathogenic bacteria, was unprecedented when compared with fourteen antibiotics and fifteen other disinfectant amphiphiles.
[0043] Given the promise of this early generation of bisQPC disinfectants, new alternatives are desired to expand this emerging class of amphiphiles. The inventors envisioned an opportunity to accomplish two parallel goals, aiming 1) to determine if the bulky phenyl groups could be replaced with smaller alkyl groups, rendering the disinfectant structures more atom economical, and 2) to prepare exact nitrogen- and phosphorus-bearing analogs in amphiphilic disinfectants, thus scrutinizing the role of the cationic element in the antimicrobial action. [0044] The inventors obtained the highly effective tetramethylethylenediamine-derived bisQACs prepared in their labs, specifically 12(2)12, and worked on comparing analogous monoQAC and monoQPC compounds. In the present disclosure, the phenyl rings on the PhP2P- n,n structure (Scheme 1, left) are replaced with alkyl substituents (Scheme 1, right), leading to series dubbed CyP2P-n,n, EtP2P-n,n, and MT2P-n,n for cyclohexyl, ethyl, and methyl analogs, respectively. Cyclohexyl-substituted QPCs would serve to inform the transition from aromatic substituents in the first generation of bisQPCs to alkyl substituents, presumably providing steric protection from air oxidation for the phosphorus atom. Preparation of methyl- and ethylsubstituted bisQPC analogs would serve to maximize atom economy within this structural motif and identify minimum structural requirements for strong bioactivity. Further, the methylsubstituted phosphonium compounds would allow for direct comparison of bisQPC to bisQAC activity (Scheme 1, bottom), since the only change in the molecules would be the pnictogen atom (nitrogen or phosphorus) of the cationic head group.
[0045] The present disclosure provides a phenyl-free biscationic quaternary phosphonium compound, an antimicrobial composition comprising such a compound, and the method of making such an antimicrobial compound or composition, and the method of using such a compound or composition for antimicrobial use. The compound or the composition provided in the disclosure has an ability to kill, prevent, or inhibit the growth of microorganisms, including but not limited to bacteria, viruses, yeast, fungi, and protozoa, or to attenuate the severity of a microbial infection. The compound or the composition provided in the disclosure has an ability to inhibit or eradicate pre-established biofilms (i.e. antibiofilm use) formed by the microorganisms.
[0046] In accordance with some embodiments, the present disclosure provides an antimicrobial composition comprising a compound having the formula
Figure imgf000013_0001
wherein: n is an integer in a range of from 2 to 20, L is a linking group being Ci-io alkyl or a cycloalkyl, R1, R2, R3, or R4 each is a C1-12 alkyl or a cycloalkyl, unsubstituted or optional substituted with a functional group selected from the group consisting of -OH, -OR’, -NH2, - NHR’, -NR’2, -SH, -SR’, -O-C(O)R’, -C(O)OR’, -C(O)R’, -CF3, -OCF3, halogen, and any combination thereof;
R’ is H or a C1-4 alkyl;
X is selected from the group consisting of a halogen, a tosylate, a hydrocarbonate, a carbonate, a sulfate, and an acetate; and m is 1 or 2.
[0047] In the present disclosure, X" is used for illustration only. It can be monovalent anions or monovalent negative ions such as fluoride ion (F“), chloride ion (Cl-), bromide ion (Br), and iodide (I-). X” may also represent multi-valent anions such as carbonate and sulfate [0048] The present disclosure provides a composition or a product described herein. In some embodiments, the antimicrobial composition is a disinfectant configured to be applied to a surface of an object in need thereof. In some embodiments, the antimicrobial composition is a pharmaceutical composition configured to be administrated to a subject in need thereof. The pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an active ingredient. In some embodiments, the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an additive for stabilizing the pharmaceutical composition.
[0049] The value of n can be in a range of from 6 to 16, from 2 to 12, from 4 to 12, from 6 to 12, from 8 to 12, from 10 to 12, or any other suitable ranges. In some embodiments, n is in a range of from 10 to 12.
[0050] In some embodiments, the linking group L is C1-6 alkyl such as -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, -CH2CH2CH2CH2CH2-, or- CH2CH2CH2CH2CH2CH2-.
[0051] In some embodiments, R1, R2, R3, or R4 each is a C1-6 alkyl or a cyclohexyl such as cyclohexyl.
[0052] X is any suitable group for anions. The value of m depends on the valence of the anions. For example, when X a halogen, a tosylate, an acetate, or a hydrocarbonate, m is 2. When X is a carbonate or a sulfate, m is 1. In some embodiments, X is a halogen selected from F, Cl, Br, and I, and m is 2. [0053] In another aspect, the present disclosure provides a compound as described herein, a method of making such a compound, and a method of using such a compound.
[0054] Examples of a suitable compound include, but are not limited to the compounds from Cl to C21 as shown below:
Figure imgf000015_0001
Figure imgf000016_0002
(C13),
Figure imgf000016_0001
Figure imgf000017_0001
[0055] In the compounds above, the counter ions may be mX" as described. For example, in some embodiments, each of these compounds comprises 2 Br", as counter ions.
[0056] In the present disclosure, the compounds are abbreviated in a format of RPLP-n,n- X. R represents R1, R2, R3, and R4, and “Me”, “Ef ’ and “Cy” represent methyl, ethyl and cyclohexyl, respectively. “L” between two P atoms represents the number of the carbon atoms in the linking group or the name of the linking group. X and n are the same as those in the formula (I). For example, Compound (Cl) having the following structure:
Figure imgf000018_0001
is coded as MT2P-8,8-Br.
[0057] In another aspect, the present disclosure provides a method of making the antimicrobial composition as described herein. In some embodiments, such a method comprises preparing the compound having the formula (I) as described. The compound having the formula [1] can be synthesized as illustrated in Schedule 2.
[0058] Scheme 2
Figure imgf000018_0002
(II) (I)
[0059] As shown in Scheme 2, the compound having the formula (I) can be prepared by the alkylation of a suitable bisphosphine starting compound having the formula (II). A suitable electrophile, bearing an alkyl or cycloalkyl backbone as well as a leaving group, can be alkylated under substitution conditions, which can optimize solvent conditions and temperature; two equivalents of the electrophile would be utilized.
[0060] The method may further comprise mixing an effective amount of the compound having the formula (I) and a carrier. Examples of a suitable carrier may include, but are not limited to, a solvent, a carrier, an additive, any other suitable ingredient, or combinations thereof. [0061] In some embodiments, the present disclosure also provides an antimicrobial composition comprising a compound having the formula (I) as described, and a carrier such as a solvent. The antimicrobial composition can also comprise other ingredients and additives. The content of the compound having the formula (I) can be in any suitable concentration. For example, in some embodiments, such a concentration can be in the range from 0.01 pM to 100 pM, for example, from 0.1 pM to 10 pM. In some embodiments, the content of the compound having the formula (I) may be at a concentration of from 0.1 wt% to 5 wt %, for example, in the range of from 0.2 wt.% to 2.5 wt. %. Examples of the carrier include but are not limited to a solvent Examples of other additives include but are not limited to surfactants, anti-foaming agents, anti-freezing agents, gelling agents, and combinations thereof.
[0062] The antimicrobial composition may also comprise a pharmaceutically acceptable carrier or excipient A pharmaceutically acceptable carrier or excipient suitable for a solid preparation such as tablets or capsules can be, for example, binders (e.g., acacia, gelatin, dextrin, hydroxypropylcellulose, methylcellulose, polyvinylpyrrolidone), solvents, dispersion media, diluents (e.g., lactose, sucrose, mannitol, com starch, potato starch, calcium phosphate, calcium citrate, crystalline cellulose), lubricants (e.g., magnesium stearate, calcium stearate, stearic acid, talc, anhydrous silicic acid), disintegrants (e.g., com starch, potato starch, carboxymethylcellulose, carboxymethylcellulose calcium, alginic acid), and wetting agents (e.g., sodium laurylsulfate).
[0063] A pharmaceutically acceptable carrier or excipient suitable for a liquid preparation, such as solutions or suspensions, can be, for example, aqueous vehicles (e.g., water), suspending agents (e.g., acacia, gelatin, methyl cellulose, carboxymethylcellulose sodium, hydroxymethyl-cellulose, aluminum stearate gel), surfactants (e.g., lecithin, sorbitan monooleate, glycerin monostearate), and non-aqueous vehicles (e.g., glycerin, propylene glycol, vegetable oil). Moreover, liquid preparations may contain preservatives (e.g., p-hydroxybenzoic acid methyl ester, p-hydroxybenzoic acid propyl ester), flavors, and/or coloring agents. The antimicrobial composition in this disclosure can be formulated to be in any suitable form, including but not limited to liquid, gel and paste.
[0064] The present disclosure also provides different products as described herein, which comprise the antimicrobial composition.
[0065] Method of Use
[0066] In another aspect, the present disclosure provides a method of killing, preventing, or inhibiting microbial growth. For example, in some embodiments, the method comprises applying the antimicrobial composition to a surface of an object in need thereof. Examples of a suitable method include but are not limited to pouring, spraying, any other suitable methods and any combinations thereof.
[0067] In some embodiments, such a method comprises administrating the antimicrobial composition to a subject in need thereof. The method of administration may include in vitro or in vivo administration. For example, the types of administration include, are not limited to, topical, oral, parenteral, rectal, intramuscular injection, inhalation, intrathecal, sublingual, nasal, transdermal, and any combination thereof.
[0068] The antimicrobial composition or the compound is used to kill, prevent, or inhibit growth of at least one group of microorganisms such as bacteria, viruses, yeast, fungi, and protozoa, or to inhibit formation of a biofilm, or disperse or eradicate a pre-established biofilm. [0069] In some embodiments, this disclosure relates to methods of treating or preventing diseases or conditions comprising administering an effective amount of a composition comprising the quaternary phosphonium compound disclosed herein to a subject in need thereof. [0070] In some embodiments, the disclosure contemplates that a quaternary phosphonium compound disclosed herein may be used in antimicrobial applications optionally in combination with other antimicrobial agents for prevention of disease onset and treatment. In some embodiments, the quaternary phosphonium compound disclosed herein may be used in medical device coatings (medical implants and tools, IV catheters), wound dressings (embedded in gauze bandages), wound rinses (i.e. surgical rinses), wound-vacuum systems, whole body baths (e.g., in combo with bleach baths for treatment of skin flares for atopic dermatitis/eczema), soaps, personal care products (body washes, lotions, soaps) for high-risk patients or for populations with high risk of exposure (e.g. athletes using common sports equipment in gym), and veterinary applications (e.g. anti-infectives for companion animals, race horses, etc.) [0071 ] In some embodiments, this disclosure relates to methods of treating or preventing a microbial infection comprising administering to a subject in need thereof an effective amount of a quaternary phosphonium compound as disclosed herein. In some embodiments, the microbial infection is a bacterial, fungal, pest, or viral infection.
[0072] In some embodiments, this disclosure relates to methods of treating or preventing bacterial infections comprising administering or contacting the skin of a subject with formula comprising a quaternary phosphonium compound as disclosed herein to a subject in need thereof. In some embodiments, the formula is administered in combination with another antibiotic agent. [0073] In some embodiments, this disclosure provides a method of using quaternary phosphonium compounds disclosed herein for treating or preventing an Acinetobacter baumannii infection, other bacterial infection, other multidrug resistant bacteria, or other microbial infection by administering an effective amount of quaternary phosphonium compounds disclosed herein to a subject in need thereof. In further embodiments, the quaternary phosphonium compound is co-administered with an antibiotic selected from the group comprising of sulfonamides, diaminopyrimidines, quinolones, beta-lactam antibiotics, cephalosporins, tetracyclines, nitrobenzene derivatives, aminoglycosides, macrolide antibiotics, polypeptide antibiotics, nitrofuran derivatives, nitroimidazoles, nicotinic acid derivatives, polyene antibiotics, imidazole derivatives or glycopeptide, cyclic lipopeptides, glycylcyclines and oxazolidinones. In further embodiments, these antibiotics include but are not limited to sulphadiazine, sulphones - [dapsone (DDS) and para-aminosalicylic (PAS)], sulfanilamide, sulfamethizole, sulfamethoxazole, sulphapyridine, trimethoprim, pyrimethamine, nalidixic acids, norfloxacin, ciprofloxacin, cinoxacin, enoxacin, gatifloxacin, gemifloxacin, grepafloxacin, levofloxacin, lomefloxacin, moxifloxacin, ofloxacin, pefloxacin, sparfloxacin, trovafloxacin, penicillins (amoxicillin, ampicillin, azlocillin, carbenicillin, cioxacillin, dicloxacillin, flucioxacillin, hetacillin, oxacillin, mezlocillin, penicillin G, penicillin V, piperacillin), cephalosporins (cefacetrile, cefadroxil, cefalexin, cephaloglycin, cefalonium, cefaloridine, cefalotin, cefapirin, cefatrizine, cefazaflur, cefazedone, cefazolin, cefradine, cefroxadine, ceftezole, cefaclor, cefonicid, ceforanide, cefprozil, cefuroxime, cefuzonam, cefmetazole, cefotetan, cefoxitin, cefcapene, cefdaloxime, cefdinir, cefditoren, cefetamet, cefixime, cefmenoxime, cefodizime, cefoperazone, cefotaxime, cefotiam, cefpimizole, cefpiramide, cefpodoxime, cefteram, ceftibuten, ceftiofur, ceftizoxime, ceftriaxone, cefoperazone, ceftazidime, cefepime), moxalactam, carbapenems (imipenem, ertapenem, meropenem) monobactams (aztreonam), oxytetracycline, chlortetracycline, clomocycline, demeclocycline, tetracycline, doxycycline, lymecycline, meclocycline, methacycline, minocycline, rolitetracycline, chloramphenicol, amikacin, gentamicin, framycetin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, telithromycin, colistin, bacitracin, tyrothricin, nitrofurantoin, furazolidone, metronidazole, tinidazole, isoniazid, pyrazinamide, ethionamide, nystatin, amphotericin-B, hamycin, miconazole, clotrimazole, ketoconazole, fluconazole, rifampicin, lincomycin, clindamycin, spectinomycin, fosfomycin, loracarbef, polymyxin B, polymyxin B Sulphate, procaine, ramoplanin, teicoplanin, vancomycin, and/or nitrofurantoin.
[0074] In some embodiments, the subject is diagnosed with a bacterial infection. In some embodiments, the subject is diagnosed with bacteremia, pneumonia, staphylococcal food poisoning, necrotizing pneumonia, necrotizing fasciitis, scalded skin syndrome, post-operation bacterial infection, medical device bacterial infection, bacterial infection of the skin, soft tissue bacterial infection, or toxic shock syndrome.
[0075] In some embodiments, this disclosure provides methods of treating or preventing a toxin-mediated bacterial infection comprising administering an effective amount of a quaternary phosphonium compound as disclosed herein to a subject in need thereof, including a subject at risk of, exhibiting symptoms of, or diagnosed with scalded skin syndrome (esp. in neonates), abscesses, necrotizing fasciitis, sepsis, or atopic dermatitis (eczema).
[0076] In some embodiments, this disclosure provides methods of treating or preventing bacterial infections or acne comprising administering to a subject in need thereof or contacting the skin of a subject in need thereof with a formula comprising of a quaternary phosphonium compound as disclosed herein. In some embodiments, the formula is administered in combination with another antibiotic.
[0077] In some embodiments, the subject is at risk of a bacterial infection due to being diagnosed with an abscess, furuncle, cellulitis, folliculitis, atopic dermatitis, psoriasis, impetigo, septic arthritis, brain abscess, bum wound, venous ulcer, diabetic foot ulcer, surgical wound, carbuncle, or meningitis.
[0078] A flow of wound rinse/irrigation solution can be applied across an open wound surface to achieve wound hydration, to remove deeper debris, and to assist with the visual examination. In some embodiments, the disclosure relates to methods of irrigating a wound using a solution comprising a quaternary phosphonium compound as disclosed herein.
[0079] In some embodiments, this disclosure provides methods of using quaternary phosphonium compounds disclosed herein for killing microbes, preventing or inhibiting microbe growth, preventing a biofilm formation, or preventing the spread of an Acinetobacter baumannii infection, other bacterial infection, other multidrug resistant bacteria, or other microbial infection by sanitizing a surface, e.g., by contacting the surface with a solid (e.g., powder), liquid, or spray composition, with a quaternary phosphonium compound disclosed herein in an effective amount. [0080] In some embodiments, this disclosure provides methods of preventing cellular infections comprising applying a quaternary phosphonium compound disclosed herein on top of or inside a cell growth medium. [0081] In some embodiments, this disclosure provides methods of preventing plant microbial infections comprising applying a quaternary phosphonium compound disclosed herein to the exterior, leaf, seed, or stem of a plant. In some embodiments, this disclosure provides methods of preventing plant microbial infections comprising applying a quaternary phosphonium compound disclosed herein on top of or into soil, dirt, sand, or other medium from which roots of the plant reside.
[0082] Compositions and Devices
[0083] In some embodiments, this disclosure provides compositions and devices comprising a quaternary phosphonium compound disclosed herein.
[0084] In some embodiments, this disclosure provides soaps and disinfectant products comprising a quaternary phosphonium compound disclosed herein. Contemplated topical formulations for skin flares (i.e., for atopic dermatitis or other infections related to a disrupted skin barrier) may be combined with another drug such as a topical steroid, anti-inflammatory agent, and promoter of skin barrier function or skin moisturizer. In some embodiments, this disclosure provides a container configured to create a liquid spray comprising a quaternary phosphonium compound disclosed herein.
[0085] In some embodiments, this disclosure provides a pharmaceutical formulation comprising a quaternary phosphonium compound disclosed herein and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical formulation is in the form of a lotion, liquid, or gel. In some embodiment, the pharmaceutical formulation is in the form of a particle, bead, tablet, capsule, pill, or injectable solution. The injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as water, dimethyl sulfoxide, mannitol, 1,3-butanediol, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.
[0086] The pharmaceutical formulation can also include any type of pharmaceutically acceptable excipients, additives, or vehicles. For example, diluents or fillers, such as dextrates, dicalcium phosphate, calcium sulphate, lactose, cellulose, kaolin, mannitol, sodium chloride, dry starch, sorbitol, sucrose, inositol, powdered sugar, bentonite, microcrystalline cellulose, or hydroxypropyl methylcellulose, may be added to the composition to increase the bulk of the composition.
[0087] In some embodiments, the formulation is a directly compressible composition comprising a quaternary phosphonium compound disclosed herein but no excipients, additives, or vehicles.
[0088] In some embodiments, the disclosure provides a pharmaceutical or cosmetic formulation comprising a quaternary phosphonium compound disclosed herein and a pharmaceutically acceptable excipient or cosmetically acceptable excipient. In some embodiments, the disclosure provides a liquid or gel formulation optionally further comprising an antibacterial agent, a topical steroid, an anti-inflammatory agent, a promoter of skin barrier function, a skin moisturizer, or combinations thereof. In some embodiments, the antibacterial agent is daptomycin, linezolid, vancomycin, nafcillin, cefazolin, dicloxacillin, clindamycin, rifampin, or sulfamethoxazole-trimethoprim (Bactrim).
[0089] In some embodiments, the disclosure provides a wound dressings or wound rinse comprising a quaternary phosphonium compound disclosed herein wherein the wound dressing comprises an absorbent pad and optionally an adhesive.
[0090] In some embodiments, the disclosure provides disinfectant sprays or wipes formulation for surfaces and fomites comprising a quaternary phosphonium compound disclosed herein.
[0091 ] In some embodiments, this disclosure provides a medical device coated with a quaternary phosphonium compound as disclosed herein. In some embodiments, the medical device is a screw, pin, plate, rod, disk, needle, catheter, tube, stent, pacemaker, defibrillators (ICDs), artificial hip or knee joint/implant, breast implant, intra-uterine device, ear tube, contact lens, or implantable pump.
[0092] In some embodiments, this disclosure provides a surgical tool coated with a quaternary phosphonium compound as disclosed herein. In some embodiments, the surgical tool is a forceps, tweezers, scalpel, knife, scissors, retractor, needle, gauze, sponge, suction, staple, stapler, clip, laparoscopic instrument, electrosurgical cauterizer, ultrasonic device, camera, camera lens, fiber optic cable, insufflator, needle, bronchoscope, cystoscope, saw, or robotic arm. [0093] In some embodiments, the disclosure provides a wound dressing comprising a quaternary phosphonium compound as disclosed herein wherein the wound dress comprises an absorbent pad and optionally an adhesive optionally in combination with another antibiotic agent. In some embodiments, the wound dressing is a foam or compression dressing or a cover dressing such as wraps, gauze and tape. In some embodiments, the wound dressing comprises alginate or collagen. In some embodiments, the wound dressing is a hydrocolloid dressing, e.g., carboxy-methylcellulose and gelatin optionally in a polyurethane foam or film, optionally comprising one or more agents selected from pectin, a polysaccharide, and an adhesive.
[0094] In some embodiments, the wound dressing is a hydrogel. Hydrogels are polymers that contain a high content of hydroxy and/or carboxyl containing monomers or salts thereof, e.g., vinyl alcohol, acrylic acid, 2-hydroxyethylmethacrylate, ethylene glycol dimethacrylate monomers, which are co-polymers to provide varying degrees of hydration. Due to the hydrophilic monomers, the hydrogels typically absorb water. Contemplated hydrogel dressings include: amorphous hydrogel, which are a free-flowing gel that are typically distributed in tubes, foil packets and spray bottles; an impregnated hydrogel, which are typically saturated onto a gauze pad, nonwoven sponge ropes and/or strips; or a sheet hydrogel which are gel held together by a fiber mesh.
[0095] In some embodiments, the disclosure provides a wound rinse comprising a quaternary phosphonium compound as disclosed herein optionally containing normal saline, sterile water, a detergent, a surfactant, a preservative, or iodine.
[0096] In some embodiments, the disclosure provides a kit comprising a container comprising a quaternary phosphonium compound as disclosed herein optionally comprising a second container comprising a rinse solution or containing surgical device or tool, normal saline, sterile water, a detergent, a surfactant, a preservative, iodine, hydrogen peroxide, or sodium hypochlorite or other compound disclosed herein.
[0097] In some embodiments, the disclosure provides a cosmetic formulation comprising a quaternary phosphonium compound as disclosed herein and cosmetically acceptable excipient or additive. In some embodiments, the disclosure relates to a solid or liquid soap or lotion comprising a quaternary phosphonium compound as disclosed herein and a fatty acid. In some embodiments, additives can be selected from the group consisting of oily bodies, surfactants, emulsifiers, fats, waxes, pearlescent waxes, bodying agents, thickeners, superfatting agents, stabilizers, polymers, silicone compounds, lecithins, phospholipids, biogenic active ingredients, deodorants, antimicrobial agents, antiperspirants, film formers, antidandruff agents, swelling agents, insect repellents, hydrotropes, solubilizers, preservatives, perfume oils and dyes. In some embodiments, additives are selected from the group consisting of surfactants, emulsifiers, fats, waxes, stabilizers, deodorants, antiperspirants, antidandruff agents, and perfume oils.
[0098] In some embodiments, this disclosure provides a cell growth medium comprising a quaternary phosphonium compound disclosed herein.
[0099] EXAMPLES
[0100] A series of biscationic quaternary phosphonium compounds, which are phenyl- free, have been prepared. Such compounds have powerful antimicrobial activities. The examples described below are for the purpose of illustration only.
[0101] 1. Exemplary Compounds and Their Syntheses
[0102] Reagents and solvents were used from Sigma- Aldrich, ThermoFisher Scientific,
Strem Chemicals, and TCI America without further purification. Reaction vials with pressure relieving septum caps were purchased from Chemglass Life Sciences. All reactions were carried out under an argon atmosphere. All yields refer to spectroscopically pure compounds. NMR spectra were measured with a 400MHz or 500MHz JEOL spectrophotometer as noted for each compound. Chemical shifts for the 3H and 13C NMR spectra were reported on a 5-scale (ppm) downfield from TMS and internally referenced to the residual chloroform-J (CDC13) signal of 7.26 ppm and 77.16 ppm respectively. 31P NMR spectra are reported against an external reference of 85% H3PO4. Coupling constants in 1H NMR spectra were calculated in Hertz. These phosphorous compounds exhibit coupling between 31P and 13C in the 13C NMR spectra consistent with a previously reported AA’X virtual coupling pattern which convolutes the experimentally observed coupling values for Jp,c with Jpp. Therefore, coupling values for these compounds have not been unambiguously determined. For all compounds, this diagnostic 31P- 13C AA’X pattern indicates the expected number of P-C bonds consistent with the indicated compound structure. Accurate mass spectrometry data was acquired on an AB Sciex 5600+ TripleTOF using electrospray ionization in positive mode.
[0103] Synthesis of alkyl bisQPCs:
[0104] General Procedure for Synthesis of bisQPCs: The appropriate 1,2- bis(dialkylphosphino)ethane was loaded to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar. Acetonitrile and the corresponding 1 -bromoalkane were added to the vial by syringe through the septum cap. The reaction vial was placed in an aluminum heating block preheated to 72-75 °C. The reaction was stirred at temperature for 5-24 h. The volatiles were removed from the vial using rotary evaporation. The products were purified either by trituration or recrystallization. All reactions were conducted under an atmosphere of argon. Detailed procedures for each compound along with all characterization data can be found in the Supporting Information.
[0105] Amphiphilic QAC and QPC compounds containing alkyl chain(s) of 10 to 12 carbon atoms display optimal efficacy as disinfectants. Therefore, a series of compounds with alkyl chain lengths between 8 and 16 carbons were prepared to bracket this hypothesized ideal nonpolar region.
[0106] Preparation of the alkyl substituted bisQPCs was readily accomplished through the treatment of the relevant bisphosphinoethane with excess equivalents of a linear chain alkyl bromide in acetonitrile at elevated temperatures (Schemes 2-3). To reduce the risk of oxidizing the alkyl phosphine, reactions were conducted under an atmosphere of argon in pressure-rated reaction vials with rupture disk septum caps. BisQPC products were generally isolated in 60 - 95% yields, with the primary source of variation in yields stemming from the process of handling and purifying waxy or glassy products. Detailed reaction procedures as well as characterization of the prepared compounds by ’H, 13C, 31P NMR and HRMS are described as follows.
[0107] Preparation of MT2P-8,8-Br
Figure imgf000027_0001
[0108] 1 ,2-Bis(dimethylphosphino)ethane (0.35 mL, 0.32 g, 2.1 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (0.7 mL) and 1 -bromooctane (1.00 mL, 1.12 g, 5.8 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 75 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 5 hours at 75 °C, and then allowed to further stir at room temperature for 18 h. The volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was triturated with diethyl ether to form a white solid. The product was isolated via vacuum filtration as a fine white powder (0.759 g, 67.5 %);1H NMR (CDC13, 500 MHz): 53.38 (br, 4H), 2.48 (br, 4H), 2.18 (t, 12H, JP,H = 5.0 Hz,), 1.53 (br, 4H), 1.42 (br quint, 4H), 1.28-1.18 (br m, 16H), 0.82 (t, 6H, JH,H = 5 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 531.74, 30.72 (t, J= 7.5 Hz), 29.09, 22.61, 22.06 (t), 21.70 (t, J= 2.5 Hz), 15.67 (quint), 14.12, 6.87 (quint); 31P{1H} NMR (CDC13, 162.0 MHz): 533.55; HRMS (ESI+): Found 188.1677, expected C22H50P2 [M-2Br-]2+ 188.1700. [0109] Preparation of MeT2P-9,9-Br
Figure imgf000028_0002
[0110] 1,2-Bis(dimethylphosphino)ethane (0.40 mL, 0.36 g, 2.4 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (0.8 mL) and 1 -bromononane (1.30 mL, 1.41 g, 6.8 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 7 hours at 72 °C, and then cooled to room temperature. The volatiles were removed from the vial using rotary evaporation. The resulting waxy, white solid was triturated with 8 mL of hexanes and the slurry was placed into a -20 °C freezer overnight. The product was isolated via vacuum filtration, then triturated again with 12 mL of diethyl ether and vacuum filtered to afford a white waxy solid. (1.26 g, 93.0 %); NMR (CDC13, 500 MHz): 53.24 (s, 4H), 2.48 (br, 4H), 2.17 (t, 12H, JP,H = 5.0 Hz,), 1.52 (br, 4H), 1.40 (br quint, 4H), 1.27-1.18 (br m, 20H), 0.80 (t, 6H, JH,H = 7.5 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 531.76, 30.68 (t, J= 8.2 Hz), 29.34, 29.18, 29.09, 22.60, 22.12 (t), 21.68 (t, J= 1.9 Hz), 15.65 (quint), 14.09, 6.92 (quint); 31P{1H} NMR (CDC13, 202.4 MHz): 533.31; HRMS (ESI+): Found 202.1834, expected C24H54P2 [M- 2Br-]2+ 202.1850.
[0111] Preparation of MeP2P-l 0,10-Br
Figure imgf000028_0001
[0112] 1,2-Bis(dimethylphosphino)ethane (0.40 mL, 0.36 g, 2.4 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (0.7 mL) and 1 -bromodecane (1.30 mL, 1.39 g, 6.3 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 75 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 5 hours at 75 °C, and then allowed to further stir at room temperature for 18 h. The volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was triturated with diethyl ether to form a white solid. The product was isolated via vacuum filtration as a fine white powder (1.41 g, 99.0%); ’H NMR (CDC13, 500 MHz): 53.32 (d, 4H, JP,H = 5.0 Hz), 2.49 (br, 4H), 2.18 (t, 12H, JP,H = 5.0 Hz,), 1.52 (br, 4H), 1.40 (br quint, 4H), 1.27-1.18 (br m, 24H), 0.80 (t, 6H, JH,H = 7.5 Hz); 13C {1H} NMR (CDC13, 125.8 MHz): 531.81, 30.69 (br t), 29.46, 29.37, 29.23, 29.08, 22.61, 22.19 (t), 21.68, 15.65 (t), 14.07, 6.95 (t); 31P{1H} NMR (CDC13, 202.4 MHz): 533.67; HRMS (ESI+): Found 216.1993, expected C26H58P2 [M]2+ 216.2000.
[0113] Preparation of MeP2P-l 1,11-Br
Figure imgf000029_0001
[0114] 1,2-Bis(dimethylphosphino)ethane (0.40 mL, 0.36 g, 2.4 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (0.6 mL) and 1 -bromoundecane (1.60 mL, 1.69 g, 7.2 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 5 hours at 72 °C, and then allowed to further stir at room temperature overnight. The volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days. The product was isolated via vacuum filtration as a fine white powder (1.335 g, 89.7%); 1H NMR (CDC13, 500 MHz): 53.33 (d, 4H, JP,H = 5.0 Hz), 2.49 (br, 4H), 2.19 (t, 12H, JP,H = 7.5 Hz,), 1.53 (br, 4H), 1.41 (br, 4H), 1.26-1.19 (br m, 28H), 0.81 (t, 6H, JH,H = 7.5 Hz); ); 13C{1H} NMR (CDC13, 125.8 MHz): 531.86, 30.77 (br t), 29.56, 29.54, 29.40, 29.30, 29.11, 22.65, 22.23 (t), 21.71, 15.70 (t), 14.10, 6.98 (t); 31P{1H} NMR (CDC13, 202.4 MHz): 533.65; HRMS (ESI+): Found 230.2151, expected C28H62P2 [M-2Br"]2+ 230.2150. [0115] Preparation of MeP2P-12,12-Br
Figure imgf000030_0001
[0116] 1,2-Bis(dimethylphosphino)ethane (0.30 mL, 0.27 g, 1.8 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (0.6 mL) and 1 -bromododecane (1.30 mL, 1.35 g, 5.4 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 5 hours at 72 °C, and then allowed to further stir at room temperature overnight. The volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days. The product was isolated via vacuum filtration as a fine white powder (0.986 g, 84.6%); 1H NMR (CDC13, 500 MHz): 53.24 (s, 4H), 2.47 (br, 4H), 2.16 (t, 12H, JP,H = 7.5 Hz,), 1.52 (br, 4H), 1.39 (br quint, 4H), 1.26-1.17 (br m, 32H), 0.80 (t, 6H, JH,H = 7.5 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 531.82, 30.68 (t, J= 8.2 Hz), 29.60, 29.58, 29.54, 29.40, 29.28, 29.10, 22.60, 22.11 (t), 21.65, 15.64 (quint), 14.05, 6.89 (quint); 31P{1H} NMR (CDC13, 202.4 MHz): 533.73; HRMS (ESI+): Found 244.2317, expected C30H66P2 [M-2Br-]2+ 244.2300.
[0117] Preparation of MT2P-14,14-Br
Figure imgf000030_0002
[0118] 1 ,2-Bis(dimethylphosphino)ethane (0.30 mL, 0.27 g, 1.8 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (0.7 mL) and 1 -bromotetradecane (1.60 mL, 1.49 g, 5.4 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 7 hours at 72 °C, and then cooled to room temperature. The volatiles were removed from the vial using rotary evaporation. The resulting waxy, white solid was triturated with 8 mL of hexanes and the slurry was placed into a -20 °C freezer overnight. The product was isolated via vacuum filtration, then triturated again with 12 mL of diethyl ether and vacuum filtered to afford a fine white powder. (1.02 g, 80.3 %); 1H NMR (CDC13, 500 MHz): 53.36 (d, 4H, JP,H = 5.0 Hz), 2.53 (br, 4H), 2.21 (t, 12H, JP,H = 5.0 Hz,), 1.57 (br, 4H), 1.47 (br quint, 4H), 1.33-1.24 (br m, 40H), 0.87 (t, 6H, JH,H = 7.5 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 532.03, 30.77 (t, J= 7.5 Hz), 29.81, 29.79, 29.77, 29.75, 29.67, 29.50, 29.48, 29.22, 22.80, 22.49 (t), 21.82, 16.03 (t), 14.24, 7.21 (quint); 3,P{,H} NMR (CDC13, 162.0 MHz): 533.74; HRMS (ESI+): Found 272.2633, expected C34H74P2 [M-2Br"]2+ 272.2650.
[0119] Preparation of MeP2P-16,16-Br
Figure imgf000031_0001
[0120] 1,2-Bis(dimethylphosphino)ethane (0.30 mL, 0.27 g, 1.8 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (0.7 mL) and 1 -bromohexadecane (1.65 mL, 1.65 g, 5.4 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 72 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 7 hours at 72 °C, and then cooled to room temperature. The volatiles were removed from the vial using rotary evaporation. The resulting waxy, white solid was triturated with 8 mL of hexanes and the slurry was placed into a -20 °C freezer overnight. The product was isolated via vacuum filtration, then triturated again with 12 mL of diethyl ether and vacuum filtered to afford a fine white powder. (0.909 g, 66.4 %); 1H NMR (CDC13, 500 MHz). 53.36 (d, 4H, JP,H = 5.0 Hz), 2.53 (br, 4H), 2.21 (t, 12H, JP,H= 5.0 Hz,), 1.57 (br, 4H), 1.47 (br quint, 4H), 1.33-1.24 (br m, 40H), 0.87 (t, 6H, JH,H = 7.5 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 531.97, 30.78 (t, J = 8.2 Hz), 29.78, 29.75, 29.72, 29.68, 29.52, 29.41, 29.22, 22.73, 22.23 (t), 21.78, 15.78 (quint), 14.18, 7.01 (quint); 31P{1H} NMR (CDC13, 202.4 MHz): 533.70; HRMS (ESI+): Found 300.2946, expected C38H82P2 [M-2Br]2+ 300.2950.
[0121] Preparation of EtP2P-8,8-Br
Figure imgf000032_0001
[0122] 1,2-Bis(diethylphosphino)ethane (0.25 mL, 0.22 g, 1.1 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (1 mL) and 1 -bromooctane (0.55 mL, 0.61 g, 3.2 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 5 h at 70 °C, then cooled to room temperature and stirred for 15 h. At the end of the stir time, the volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 1 day. The product was isolated via vacuum filtration as a fine white powder (0.404 g, 63.8%); NMR (CDC13, 500 MHz): 53.29 (d, 4H, JP,H = 5.0 Hz), 2.65 (br sextet, 8H), 2.62 (br q, 4H), 1.55 (br, 4H), 1.45 (br quint, 4H), 1.33-1.20 (br m, 28H), 0.81 (t, 6H, JH,H = 1.5 HZ); 13C{1H} NMR (CDC13, 125.8 MHz): 531.66, 30.85, 28.99, 22.57, 21.88 (br t), 18.53 (quint), 14.06, 12.69 (t), 6.37-6.25 (m, two signals); 31P{1H} NMR (CDC13, 202.4 MHz): 540.14; HRMS (ESI+): Found 216.1996, expected C26H58P2 [M- 2Br"]2+ 216.2000.
[0123] Preparation of EtP2P-10,10-Br
Figure imgf000032_0002
[0124] 1 ,2-Bis(diethylphosphino)ethane (0.28 mL, 0.25 g, 1.2 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (1 mL) and 1 -bromodecane (0.75 mL, 0.80 g, 3.6 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 6 h at 75 °C, then cooled to room temperature. The volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days. The product was isolated via vacuum filtration as a fine white powder (0.689 g, 87.6%); 1H NMR (CDC13, 500 MHz): 53.15 (d, 4H JP,H = 10.0 Hz), 2.56 (br sextet, 8H), 2.48 (br, 4H), 1.51 (br, 4H), 1.41 (br quint, 4H), 1.28-1.16 (br m, 36H), 0.78 (t, 6H, JH,H = 7.5 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 531.75, 30.77, 29.39, 29.30, 29.17, 28.97, 22.55, 21.76. 18.24 (t), 14.01, 12.40 (t), 6.29-6.14 (m); 31P{1H} NMR (CDC13, 202.4 MHz): 540.40; HRMS (ESI+): Found 244.2311, expected C30H66P2 [M-2Br‘]2+ 244.2300.
[0125] Preparation of EtP2P-l 1,11-Br
Figure imgf000033_0001
[0126] 1,2-Bis(diethylphosphino)ethane (0.28 mL, 0.25 g, 1.2 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (1 mL) and 1 -bromodecane (0.75 mL, 0.80 g, 3.6 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 6 h at 75 °C, then cooled to room temperature. The volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days. The product was isolated via vacuum filtration as a fine white powder (0.488 g, 59.4%); 1H NMR (CDC13, 500 MHz): 53.26 (d, 4H, JP,H = 10.0 Hz), 2.61 (br sextet, 8H), 2.51 (br q, 4H), 1.54 (br, 4H), 1.44 (br quint, 4H), 1.32-1.18 (br m, 40H), 0.81 (t, 6H, JH,H = 7.5 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 531.83, 30.84, 29.53, 29.48, 29.34, 29.27, 29.03, 22.64, 21.87 (t), 18.48 (t), 14.09, 12.63 (t), 6.34-6.26 (m, two signals); 31P{1H} NMR (CDC13, 202.4 MHz): 540.20; HRMS (ESI+): Found 258.2476, expected C32H70P2 [M-2Br]2+ 258.2500.
[0127] Preparation of EtP2P-12,12-Br
Figure imgf000034_0001
[0128] 1,2-Bis(diethylphosphino)ethane (0.28 mL, 0.25 g, 1.2 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (1 mL) and 1 -bromododecane (0.87 mL, 0.90 g, 3.6 mmol) were added to the vial, also by syringe. The reaction vial was placed in an aluminum heating block preheated to 75 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 6 h at 75 °C, then cooled to room temperature. The volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 2 days. The product was isolated via vacuum filtration as a fine white powder (0.539 g, 63.7%); NMR (CDC13, 500 MHz): 53.17 (br, 4H), 2.60 (br sextet, 8H), 2.49 (br, 4H), 1.54 (br, 4H), 1.44 (br, 4H), 1.31-1.18 (br m, 44H), 0.81 (t, 6H, JH,H = 7.5 Hz); “CfH} NMR(CDC13, 125.8 MHz): 531.87, 30.83, 29.61, 29.59, 29.52, 29.38, 29.31, 29.05, 22.65, 21.85, 18.30 (t), 14.10, 12.50 (t), 6.31-6.21 (m, two signals); 31P{1H} NMR (CDC13, 202.4 MHz): 540.37; HRMS (ESI+): Found 272.2624, expected C34H74P2 [M-2Br-]2+ 272.2650. [0129] Preparation of ElP2P-14,14-Br
Figure imgf000034_0002
[0130] 1,2-Bis(diethylphosphino)ethane (0.26 mL, 0.23 g, 1.1 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (1 mL) and 1 -bromotetradecane (1.0 mL, 0.93 g, 3.4 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 5 h at 70 °C, then cooled to room temperature and stirred for 15 h. At the end of the stir time, the volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 1 day. The product was isolated via vacuum filtration as a fine white powder (0.744 g, 87.7 %); 1H NMR (CDC13, 500 MHz): 53.22 (br, 4H), 2.60 (br, 8H), 2.50 (hr, 4H), 1.54 (br, 4H), 1.43 (br , 4H), 1.32-1.19 (br m, 52H), 0.82 (t, 6H, JH,H = 7.5 Hz); 13C{1H} NMR(CDC13, 125.8 MHz): 531.85, 30.82, 29.61, 29.58, 29.50, 29.35, 29.29, 29.03, 22.62, 21.82, 18.32 (t), 14.07, 12.49 (t), 6.27-6.19 (m, two signals); 31P{1H} NMR (CDC13, 202.4 MHz): 540.29; HRMS (ESI+): Found 300.2931, expected C38H74P2 [M-2Br"]2+ 300.2950.
[0131] Preparation of EtP2P-16,16-Br
©
© P/
2Br°
[0132] 1,2-Bis(diethylphosphino)ethane (0.26 mL, 0.23 g, 1.1 mmol) was loaded via syringe to a 40 mL reaction vial with pressure relieving septum cap which contained a magnetic stir bar and was flushed with argon for 15 minutes prior to the addition. Acetonitrile (1 mL) and 1 -bromohexadecane (1.0 mL, 1.0 g, 3.3 mmol) were added to the vial also by syringe. The reaction vial was placed in an aluminum heating block preheated to 70 °C, venting with a needle twice during the first 5 minutes to avoid over pressurization of the vial. The reaction was stirred for 5 h at 70 °C, then cooled to room temperature and stirred for 15 h. At the end of the stir time, the volatiles were removed from the vial using rotary evaporation. The resulting colorless, viscous oil was dissolved into ~2 mL of dichloromethane, layered with ~30 mL hexanes and placed into a -20 °C freezer. After 3 h in the freezer, the vial was swirled to help initiate the formation of a white solid, then was allowed to sit in the freezer for 1 day. The product was isolated via vacuum filtration as a fine white powder (0.870 g, 95.5 %); 1H NMR (CDC13, 500 MHz): 53.22 (br, 4H), 2.60 (br, 8H), 2.51 (br, 4H), 1.55 (br, 4H), 1.45 (br, 4H), 1.32-1.19 (br m, 60H), 0.82 (t, 6H, JH,H = 5.0 Hz); 13C{1H} NMR (CDC13, 125.8 MHz): 531.92, 30.87 (br t), 29.71, 29.68, 29.65, 29.56, 29.40, 29.36, 29.08, 22.69, 21.89, 18.43 (t), 14.13, 12.59 (t), 6.35- 6.25 (m, two signals); 31P{1H} NMR (CDC13, 202.4 MHz): 540.30; HRMS (ESI+): Found 328.3245, expected C42H90P2 [M-2Br]2+ 328.3250.
[0133] Preparation of CyP2P-8,8-Br
Figure imgf000036_0001
[0134] 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromooctane (0.18 mL, 0.20 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer. After two days, the mother liquor was decanted from the oil that formed on the bottom of the vial. Residual solvent was again removed by rotary evaporation, resulting in the product as a white, glassy solid. (0.375 g, 94.6 %); 1H NMR (CDC13, 500 MHz), 53.09 (d, J= 5.3 H, 4H) 3.06-2.93 (m, 4H) 2.58 (m, 4H) 2.07 - 1.66 (m, 20H) 1.56 - 1.38 (m, 24H) 1.30 - 1.13 (m, 20H) 0.79 (t, J - 6.8 Hz, 6H) ; 13C{1H} NMR (CDC13, 125.8 MHz) 5 31.72, 31.06 (t, J= 7.1 Hz) 30.68 (t, J= 49.06) 29.08, 29.03, 27.02, 26.01 (t, J= 6.2 Hz), 25.34, 22.72 (t, J = 2.2 Hz), 22.63, 16.51, 14.01, 11.83 31P{1H} NMR (CDC13, 202.5 MHz>, 536.88. HRMS (ESI+): Found 324.2931, expected m/z C42H82P2 [M-2Br]2+ 324.2941.
[0135] Preparation of CyP2P-9,9-Br
Figure imgf000036_0002
[0136] 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromononane (0.20 mL, 0.22 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by loosening the cap twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white powder was purified via trituration with diethyl ether (approx. 30 mL) and isolated by vacuum filtration, resulting in the product as a white solid. (0.387 g, 92.4 %); 1H NMR (CDC13, 500 MHz), 83.19 (d, J= 5.5 H, 4H) 3.04 - 2.97 (m, 4H) 2.62 - 2.55 (m, 4H) 2.07 - 1.70 (m, 20H) 1.60 - 1.34 (m, 24H) 1.35 - 1.10 (m, 25H) 0.80 (t, J = 6.9 Hz, 6H) ; 13C{1H} NMR (CDC13. 125.8 MHz) 5 31.85, 31.07 (t, J= 6.3 Hz) 30.68 (t, J= 20.1) 29.34, 29.21, 29.16, 27.12, 26.10 (t, J= 6.3 Hz), 25.44, 22.74 (t, J= 2.5 Hz), 22.67, 16.53, 14.13, 11.8731P{1H} NMR (CDC13, 202.5 MHz), δ 36.87. HRMS (ESI+): Found 338.3095, expected m/z C44H86P2 [M-2Br]2+ 338.3097. [0137] Preparation of CyP2P- 10,10-Br
Figure imgf000037_0001
[0138] 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromodecane (0.22 mL, 0.23 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was dissolved in (approx. 2 mL) chloroform-d layered with (approx. 30 mL) hexanes and place in a -20 °C freezer. The resulting white powder was isolated by vacuum filtration, resulting in the product as a white solid. (0.126 g, 29.2 %); 1H NMR (CDC13, 500 MHz), 53.09 (d, J= 5.0 Hz, 4H) 3.02 - 3.00 (m, 4H) 2.61 -2.57 (m, 4H) 2.04 - 1.74 (m, 18H) 1.56 - 1.45 (m, 22H) 1.28 - 1.20 (m, 26H) 0.81 (t, J - 5.0 Hz, 6H) ; 13C{1H} NMR (CDC13, 125.8 MHz) 531.82, 30.99 (t, J = 6.3 Hz) 30.60 (t, J= 21.4) 29.46, 29.33, 29.26, 29.10, 27.05, 26.03 (t, J= 6.3 Hz), 25.38, 22.65, 22.63, 16.38, 14.08, 11.68 ; 31P{1H} NMR (CDC13, 202.5 MHz;, 536.94. HRMS (ESI+): Found 352.3243, expected m/z C46H90P2 [M-2Br]2* 352.3254.
[0139] Preparation of ^P-l 1,11-Br
Figure imgf000038_0001
[0140] 1 ,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromoundecane (0.23 mL, 0.25 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C vented by puncturing the cap with a needle twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer. After five days, the mother liquor was decanted from the oil that formed on the bottom of the vial. Residual solvent was again removed by rotary evaporation, resulting in the product as a white, glassy solid. (0.324 g, 72.5 %); 1H NMR (CDC13, 400 MHz), 53.12 (d, J= 5.5 Hz, 4H) 3.06 - 2.96 (m, 4H) 2.62 - 2.54 (m, 4H) 2.04 - 1.73 (m, 20H) 1.59 - 1.40 (m, 24H) 1.30 - 1.13 (m, 32H) 0.80 (t, J - 6.8 Hz, 6H) ; 13C{1H} NMR (CDC13. 100.6 MHz) 531.91, 31.07 (t, J= 8.8 Hz) 30.67 (t, J= 25.1) 29.62, 29.56, 29.40, 29.33, 29.17, 27.12, 26.10 (t, J= 7.5 Hz), 25.43, 22.77, 22.71, 16.52, 14.16, 11.8631P{1H} NMR(CDC13, 162.0 MHz/ 536.87. HRMS (ESI+): Found 366.3384, expected m/z C48H94P2 [M-2Br]2+ 366.3410. [0141] Preparation of CyP2P- 12,12-Br
Figure imgf000039_0001
[0142] 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromododecane (0.25 mL, 0.26 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer. After two days, the mother liquor was decanted from the oil that formed on the bottom of the vial. Residual solvent was again removed by rotary evaporation, resulting in the product as a white, glassy solid. (0.334 g, 72.5 %); 1H NMR (CDCI3, 400 MHz), δ 3.11 (d, J= 4 Hz, 4H) 3.07 - 2.97 (m, 4H) 2.62 - 2.55 (m, 4H) 2.05 - 1.74 (m, 20H) 1.60 - 1.40 (m, 24H) 1.32 - 1.16 (m, 36H) 0.82 (t, J - 4 Hz, 6H) ; 13C{1H} NMR (CDC13, 100.6 MHz) δ 31.94, 31.08 (t, J= 8.8 Hz) 30.68 (t, J = 25.1) 29.68, 29.64, 29.58, 29.41, 29.37, 29.18, 27.12, 26.11 (t, J= 6.0 Hz), 25.44, 22.74, 22.72, 16.30, 14.17, 11.6431P{1H} NMR (CDC13, 162.0 MHz/ δ 36.88. HRMS (ESI+): Found 380.3553, expected m/z C50H98P2 [M- 2Br]2+ 380.3567.
[0143] Preparation of CyP2P-13,13-Br
Figure imgf000040_0001
[0144] 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromotridecane (0.27 mL, 0.28 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was washed with diethyl ether. Residual solvent was again removed by rotary evaporation, resulting in the product as a white, glassy solid. (0.401 g, 84.6 %); 1H NMR (CDC13, 500 MHz), 53.18 (d, J= 5.0 Hz, 4H) 3.11 - 3.02 (m, 4H) 2.67 - 2.62 (m, 4H) 2.10 - 1.74 (m, 26H) 1.62 - 1.48 (m, 24H) 1.34 - 1.24 (m, 42H) 0.87 (t, J - 5.0 Hz, 6H) ; 13C{1H} NMR (CDC13, 125.8 MHz) 532.04, 31.16 (t, .7= 7.5 Hz) 30.78 (t, .7= 20.2) 29.78, 29.76, 29.67, 29.50, 29.47, 29.27, 27.21, 26.20 (t, J = 6.3 Hz), 25.53, 22.86, 22.81, 16.63, 14.25, 11.82 ^PfH} NMR (CDC13, 202.5 MHz) δ 36.88. HRMS (ESI+): Found 380.3553, expected m/z C50H98P2 [M-2Br]2+ 380.3567.HRMS (ESI+): Found 394.3693, expected m/z C52H102P2 [M- 2Br]2- 394.3723.
[0145] Preparation of CyP2P-14,14-Br
Figure imgf000040_0002
[0146] 1,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromotetradecane (0.31 mL, 0.29 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer. After two days, the mother liquor was decanted from the oil that formed on the bottom of the vial. Residual solvent was again removed by rotary evaporation, resulting in the product as a white, glassy solid. (0.379 g, 77.6 %); 1H NMR (CDC13, 500 MHz), 53.12 (d, J= 4.0, 4H) 3.06 - 2.98 (m, 4H) 2.62 - 2.56 (m, 4H) 2.05 - 1.75 (m, 20H) 1.57 - 1.43 (m, 24H) 1.30 - 1.20 (m, 46H) 0.82 (t, J= 4.0, 6H) ; 13C{1H} NMR (CDC13, 125.8 MHz) 531.92, 31.03 (t, J= 8.8 Hz) 30.65 (t, J= 25.1) 29.68, 29.66, 29.65, 29.55, 29.38, 29.36, 29.14, 27.09, 26.07 (t, J= 6.0 Hz), 25.41, 22.72, 22.69, 16.47, 14.11, 11.80 31P{1H} NMR (CDC13, 202.5 MHz), 536.89. HRMS (ESI+): Found 408.3862, expected m/z C54H106P2 [M-2Br]2+ 408.3880.
[0147] Preparation of CyP2P- 16,16-Br
Figure imgf000041_0001
[0148] 1 ,2-Bis(dicyclohexylphosphino)ethane (0.211 g, 0.5 mmol) was massed into a 40 mL reaction vial with pressure relieving septa cap equipped with a stir bar which was preliminarily flushed with argon for 15 minutes. After the addition of 1,2- bis(dicyclohexylphosphino)ethane, the reaction vial was flushed with argon for an additional 10 minutes. Acetonitrile (1 mL) and 1 -bromohexadecane (0.32 mL, 0.32 g, 1.0 mmol) were added to the reaction vial via needle and syringe. The reaction vial was placed in an aluminum heating block which was preheated to 72 °C and vented by puncturing the cap with a needle twice to relieve pressure buildup. The reaction vial was stirred at 72 °C for 24 hours. Within 45 minutes, the reaction contents changed from a slurry to a clear solution and remained as a solution through the entire reaction time. At the end of 24 hours, the reaction vial was removed from heat and cooled to room temperature. All volatiles were removed by rotary evaporation. The resulting white gel was dissolved in (approx. 2 mL) dichloromethane layered with (approx. 30 mL) hexanes and place in a -20 °C freezer. After five days, the mother liquor was decanted from the oil that formed on the bottom of the vial. Residual solvent was again removed by rotary evaporation. Finally, the white, glassy solid was washed with diethyl ether and isolated by vacuum filtration. (0.377 g, 73.0 %); 1H NMR (CDC13, 400 MHz), δ 3.10 (d, J = 4.0 Hz, 4H) 3.03 - 2.96 (m, 4H) 2.60 - 2.54 (m, 4H) 2.03 - 1.72 (m, 20H) 1.55 - 1.43 (m, 22H) 1.26 - 1.17 (m, 52H) 0.79 (t, J= 4.0 Hz, 6H) ; 13C{1H} NMR (CDC13, 100.6 MHz) 531.85, 30.96 (t, J= 6.0 Hz) 30.57 (t, J= 16.1 Hz) 29.63, 29.61, 29.59, 29.57, 29.47, 29.30, 29.28, 29.06, 26.99, 25.99 (t, J= 5.0 Hz), 25.33, 22.64, 22.61, 16.40, 14.05, 11.75 31P{1H} NMR (CDC13, 162.0 MHz), δ 36.87. HRMS (ESI+): Found 436.418, expected m/z C58H114P2 [M-2Br]2+ 436.419.
[0149] For the results of the Examples, the proton and carbon NMR signals can be corrected with each other in one plot to identify the spectroscopically unique methyl groups in the molecule and all methylene signals.
[0150] The structures and the yields of the Examples are summarized in Scheme 3.
[0151] Scheme 3
Figure imgf000043_0001
[0152] The sterically hindered 1 ,2(bisdicyclohexylphosphonium)ethane was slow to oxidize under ambient atmospheric conditions allowing for quick handling of the solid on the bench. Subsequent exclusion of air from the reaction vial via a 15-minute argon flush following the addition of the phosphine created a suitable reaction environment. The more reactive l,2(bisdiethylphosphonium)ethane and l,2(bisdimethylphosphonium)ethane were significantly more air sensitive, and therefore were loaded to argon-flushed reaction vials via syringe through a septum cap using standard Schlenk line techniques. The quantity of acetonitrile used as a solvent for all reactions was minimized to reduce adventitious water. These measures effectively prevented the formation of phosphine oxide compounds, as evidenced the observation of little to no oxidized product being observed in 31P NMR spectra, even in crude product mixtures.
[0153] The reaction times required to reach conversion correlated to the expected influence of steric hindrance around the phosphine. The less sterically constrained methyl- and ethyl-substituted phosphines quickly increased viscosity upon heating and the reactions were complete within 5-6 hours, while the phosphines with the much larger cyclohexyl groups generally required heating for 24 hours to ensure complete alkylation. It is notable that, while reactions with alkyl bromides were uniformly successful, the use of alkyl chloride electrophiles led to incomplete reactions and, ultimately, significant phosphorus oxidation.
[0154] Despite this observation, the counterions for the compounds provided in the present disclosure is not limited to bromide only. The counterions may include, but are not limited to, F", Cl", Br", I", tosylate, carbonate, sulfate, and any combination thereof. In some embodiments, an exemplary compound with counterions different from Br" may be synthesized using the schemes described with Br- counterions. In some embodiments, Br" counterions in an exemplary compound may be replaced by other counter ions after the synthesis of such a compound with Br- counterions.
[0155] 2. Testing methods:
[0156] Dynamic Surface Tension Measurements
[0157] All samples were analyzed using a Kruss BPT Mobile bubble pressure tensiometer. Solutions of 12(2)12 and MT2P-12,12 were made in concentrations of 100, 300 and 1000 ppm using deionized water. Measurements were conducted at 22 ± 1 °C. Before each sample run, a new capillary tip was installed and the instrument was calibrated with deionized water. The dynamic surface tension (mN/tn) was measured as a function of surface bubble age (30 collection points over the range of 10 to 10,000 ms).
[0158] Biological Assays
[0159] Biological testing procedures were adapted from prior work by Melander and coworkers. For all biological assays, laboratory strains of methicillin-susceptible Staphylococcus aureus MSSA (SH1000), Staphylococcus aureus CA-MRSA (USA300-0114), Staphylococcus aureus HA-MRSA (ATCC 33591), Escherichia coli (MC4100), Enterococcus faecalis (OG1RF), Acinetobacter baumannii (ATCC 17978), were grown with shaking at 37 °C overnight from freezer stocks in 5 mL of the indicated media: SH1000, OG1RF, MC4100, USA300-0114, and ATCC 17978 were grown in BD™ Mueller-Hinton broth (MHB), whereas ATCC 33591 was grown in BD™ tryptic soy broth (TSB). The inoculum were then streaked onto Mueller-Hinton agar (MHA) plates and grown overnight at 37 °C. From the plates, single colonies were selected, diluted into 50 mL of fresh media and grown overnight at 37 °C with shaking for use in the MIC assays. Optical density (OD) measurements were obtained using a SpectraMax iD3 plate reader (Molecular Devices, United States).
[0160] Minimum Inhibitory Concentration (MIC) [0161] Compounds were serially diluted two-fold from stock solutions (1.0 mM) to yield twelve 100 pL test concentrations, wherein the starting concentration of dimethyl sulfoxide (DMSO) was 2.5%. Overnight 5. aureus, E.faecalis, E. coli, A. baumannii, USA300-0114 (CA- MRSA), and ATCC 33591 (HA-MRSA) cultures were diluted to ca. 106 CFU/mL in MHB or TSB and regrown to mid-exponential phase, as determined by optical density recorded at 600 nm (ODeoo). All cultures were then diluted again to ca. 106 CFU/mL and 100 pL were inoculated into each well of a U-bottom 96- well plate (Coming, 351177) containing 100 pL of compound solution. Plates were incubated statically at 37 °C for 72 hours upon which wells were evaluated visually for bacterial growth. The MIC was determined as the lowest concentration of compound resulting in no bacterial growth visible to the naked eye, based on the highest value in three independent experiments. Aqueous DMSO controls were conducted as appropriate for each compound.
[0162] Red Blood Cell (RBC) Lysis Assay (lysisio)
[0163] RBC lysis assays were performed on mechanically defibrinated sheep blood (Hemostat Labs: DSB030). An aliquot of 1.5 mL blood was placed into a microcentrifuge tube and centrifuged at 3,800 rpm for ten minutes. The supernatant was removed, and the cells were resuspended with 1 mL of phosphate-buffered saline (PBS). The suspension was centrifuged as described above, the supernatant was removed, and cells were resuspended 4 additional times in 1 mL PBS. The final cell suspension was diluted twentyfold with PBS. Compounds were serially diluted with PBS two-fold from stock solutions (1.0 mM) to yield 100 pL of twelve test concentrations on a flat-bottom 96- well plate (Coming, 351172), wherein the starting concentration of DMSO was 2.5%. To each of the wells, 100 pL of the twentyfold suspension dilution was then inoculated. TritonX (1% by volume) served as a positive control (100% lysis marker) and sterile PBS served as a negative control (0% lysis marker). Samples were then placed in an incubator at 37 °C and shaken at 200 rpm. After 1 hour, the samples were centrifuged at 3,800 rpm for ten minutes. The absorbance of the supernatant was measured with a UV spectrometer at a 540 nm wavelength. The concentration inducing 20% RBC lysis was then calculated for each compound based upon the absorbances of the TritonX and PBS controls. Aqueous DMSO controls were conducted as appropriate for each compound.
[0164] 3. Characterization data [0165] In comparing the spectroscopic data of MT2P-12,12 to its bisQAC analog 12(2)12, significant differentiation was noted. Both the 1H NMR and 13C NMR data demonstrate a marked upfield shift in the signals for atoms neighboring the phosphonium cation, as compared to the ammonium cation as shown in Scheme 4. Scheme 4 shows the characterization data for bisQAC and bisQPC compounds 12(2)12 and MeP2P-12,12. The NMR chemical shifts are shown.
[0166] Scheme 4.
Figure imgf000046_0001
[0167] Comparison of the methyl substituents was particularly revealing, as chemical shifts for the 1H NMR signals assigned to the methyl substituents on the phosphonium cation were 2.2 ppm compared to the corresponding methyl groups on the ammonium cation of 3.4 ppm, indicating a more electron rich environment on the methyl substituents adjacent to the phosphonium cation. It was also reported P-CH3 1H NMR signals near 2 ppm in monoQPCs. An 1H-13C HSQC NMR experiment was subsequently conducted on each of these compounds to unambiguously identify the 13C NMR signal associated with the methyl group within each compound. From this analysis, the methyl carbons again revealed a marked change, from a chemical shift of 51.3 ppm in 12(2)12 to a significantly more shielded 6.9 ppm for MeP2P-12, 12.. Similar spectra were reported in ionic liquid investigations; chemical shift differences were rationalized by electronegativity differences of nitrogen versus phosphorus when bonded to carbon atoms at the amphiphilic head group. This result highlights the markedly different electronic environments and cationic distribution in these pnictogen analogs.
[0168] In order to further investigate these highly analogous bis-cationic structures, a Kruss bubble pressure tensiometer was utilized to measure the dynamic surface tension of 12(2)12 and MT2P-12,12. [0169] Antimicrobial activity is expected to be affected by differences in solution behavior of amphiphilic molecules, such as the propensity to form micelles and solution dispersion rates. Structural factors such as nature of the charge and the alkyl chain length of the amphiphile have been previously reported to change the behavior of surfactants. These types of structural factors are consistent with the factors that influence the inventors’ proposed mechanism of action for bacteria membrane disruption by multicationic QACs. The inventors, therefore, were interested if the identity of the cationic species in these amphiphiles would change dynamic surface tension as a physical property that could be correlated to observed bioactivity data. The dynamic surface tension experiments were completed at three difference concentrations (100, 300 and 1000 ppm) to elucidate any concentration dependance upon the data.
[0170] FIG. 1 shows characterization data for bisQAC and bisQPC compounds 12(2)12 and MeP2P-12,12, including dynamic surface tension (mN/m) data when plotted as a function of surface age (ms) at concentrations of 100, 300 and 1000 ppm. Bromide counter ions omitted for clarity.
[0171] When comparing the dynamic surface tension of 12(2)12 and NfcP2P-12,12, it is evident that these compounds behave almost identically in solution, despite the change of cationic center (FIG. 1). This indicates the similarity in structure may be the more prevalent factor in antimicrobial behavior than the nature of the cationic head itself for these analogous compounds.
[0172] 4. Biological activity
[0173] The prepared library of biscationic amphiphilic compounds was assessed for performance as antibacterial disinfectants by determining the minimum inhibitory concentration (MIC) for each against a panel of bacteria. The bacteria selected for the study were chosen to evaluate the broad spectrum antimicrobial performance of the QPCs, and therefore, included both Gram-positive and Gram-negative bacterial strains, including those with known antimicrobial resistance mechanisms. These bacteria include the Gram-positive strains of methicillin-susceptible Staphylococcus aureus [MSSA; SHI 000], community-acquired methicillin-resistant Staphylococcus, aureus [CA-MRSA; USA 300-0114], hospital-acquired methicillin-resistant Staphylococcus aureus [HA-MRSA; ATCC 33591] and the Gram-negative strains of Enterococcus faecalis [OG1RF], Escherichia coli [MC4100], Acinetobacter baumannii [ATCC 17498] and Pseudomonas aeruginosa [PAO1].
[0174] The potential cytotoxicity of the compounds was also considered with the evaluation of hemolysis (lysis20) assays with 20% lysis of red blood cells serving as an indicator of potentially toxic compounds. Finally, the bioactivity of these compounds were benchmarked against a commercial benzalkonium chloride source (BAG; 70% benzyldimethyldodecylammonium chloride, and 30% benzyldimethyltetradecylammonium chloride) as well as some previously reported disinfectant compounds from the inventors’ groups; included were bisQPC PhP6P-10,10 as well as PhP2P-10,10 and 12(2)12. A detailed description of the experimental procedures used to evaluate bioactivity and hemolysis assays are described above. The resulting data are summarized in Table 1.
[0175] Table 1 summarizes biological activity and lysis20 data of alkyl bisQPC compounds.
[0176] Table 1.
MIC (|N)
Figure imgf000049_0001
[0177] While the physical characterization of the bisQPC structures in the present disclosure demonstrated some similarities and differences in comparison to the nitrogenous bisQ AC analogs, the inventors were most interested in biological activity assessments, particularly in comparison to phenyl-containing analogs. The most visible bioactivity trend indicated that the highest antimicrobial activity was observed for the structures with alkyl chain lengths of 10-12 carbons. Performance progressively diminished as chain lengths either increased to 16 carbons (compromising solubility) or decreased to 8 carbons (reducing amphiphilicity). The most effective disinfectants within this series were observed to be CyP2P- 11,11, EtP2P-12,12 and MeP2P-12,12; all display single-digit micromolar activity with just one exception in the EtP2P- 12, 12 panel. [0178] This correlation between chain length and bioactivity has been observed. Among these three lead compounds, CyP2P-11,11 has a slight overall advantage in broad-spectrum potency within this data set, with MIC values of 1-8 μM All three of these top compounds clearly out-perform the commercial BAC control, but do not reach the level of performance observed with a preferred QPC, PhP6P-l 0, 10, with MIC values of 1 -4 μM Notably, the topperforming compounds still maintained high efficacy against the Gram-negative pathogens, which bear known disinfectant resistance mechanisms, such as the expression of multidrug efflux pumps with cross-resistance to QACs. Additionally, relative to BAC, improved activity was observed against the Gram-negative pathogens HA-MRSA and E. faecalis, also known to harbor multidrug efflux pumps that decrease their susceptibility to disinfectants. Together, these potent bioactivities against prominent refractory Gram-positive and Gram-negative pathogens underscore the promise of QPCs as a nascent, structurally diverse disinfectant class that can evade rapidly spreading QAC resistance.
[0179] In comparison of this set to structurally analogous phenyl-substituted PhP2P- 10,10, the bioactivities of these top three compounds approaches parity. Interestingly, an additive effect seems to be present between the alkyl chain length (n) and the substituent (R) on the compound. When the substituent is Ph or Cy, the optimum alkyl chain length leans to the shorter decyl and undecyl chains (n = 10 and 11), while for Et and Me substitutions the dodecyl chains (n = 12) are more potent.
[0180] The similarities in performance between PhP2P-10,10 and CyP2P- 11,11 suggest that any electronic differences that may be introduced in the change from the aromatic and electron withdrawing nature of phenyl rings to the most electron donating of the alkyl substituents, cyclohexyl, do not lead to a marked change in bioactivity. While potential explanations such as improved directionality of the alkyl chain due to the steric constraint of the larger groups, changes in lipophilicity and micellular formation of compounds containing larger substituent groups on the phosphonium atoms, and/or the ability of larger substituents to evade efflux pump resistance mechanisms could be proposed as influencing factors towards this empirical observation, further studies would be needed to confirm such. In consideration of the overall subtle differences in bioactivity between PhP2P-l 0,10, CyP2P-ll,ll, EtP2P-12,12 and MeP2P-12,12, it can be concluded that any of these phosphorus substituents can lead to promising disinfectants. In situations when atom economy is paramount, methyl substituted phosphonium 6ations can serve well while enjoying a decrease in molecular weight of -200 g/mol compared to the phenyl- or -260 g/mol compared to the cyclohexyl-substituted counterparts.
[0181] Within this series, a direct comparison is made between a structurally analogous bisQAC and bisQPC, 12(2)12 and MeP2P-12,12, have the same architecture, only differing in the ammonium or phosphonium cationic center. Interestingly, despite this difference in how the phosphonium and ammonium cations distribute charges, these two compounds have almost identical bioactivity across the bacterial panel, which parallels the similarity observed in the dynamic surface tension measurements. This comparable bioactivity would imply the structural feature of alkyl chain length has a much greater influence on the MIC value than the nature of the polar head group in comparing these two compounds. Research is being done to identify and evaluate these similarities and differences in electronic and structural features of QPCs and QACs and the influence on their performance as disinfectants and propensity toward resistance mechanisms.
[0182] 5. Biofilm Eradication Procedure
[0183] Biofilm eradication experiments were performed using a pegged-lid microtiter plate assay to determine the MBEC values for compounds of interest, as previously described. See Yang, H; Abouelhassan, ¥.; Burch, G.M.; Kallifidas, D.; Huang, G; Yousaf, H; Jin,S.; Luesch, H.; Huigens, RW. A Highly Potent Class of Halogenated Phenazine Antibacterial and Biofilm-Eradicating Agents Accessed Through a Modular Wohl-Aue Synthesis. Sei. Rep. 2017, 7, 2003; andRaval, Y. S., Flurin, L., Mohamed, A., Greenwood-Quaintance, K. E., Beyenal, H, & Patel, R. In Vitro Activity of Hydrogen Peroxide and Hypochlorous Acid Generated by Electrochemical Scaffolds Against Planktonic and Biofilm Bacteria. Antimicrob Agents Chemother. 2021, 65, eOl 966-20. Briefly, 125 mL of mid-log phase culture diluted to ca. 10 6 CFU/mL in tryptic soy broth (TSB) was added to wells of plat-bottom 96- well plates (Thermo Scientific, 266120) and covered with 96-pegged lid (Thermo Scientific, 445497). Plates were incubated to establish bacterial biofilms after incubation at 37 °C for 24 hours. The pegged lid was then removed, washed with PBS, and transferred to another 96-well plate containing 2-fold serial dilutions of the test compounds (the “challenge plate”). The total volume in each well was 150 mL, comprising of 75 mL of compound diluted in water/DMSO, with a starting DMSO concentration of 2.5 %, and 75 mL of TSB. Plates were incubated statically at 37 °C for 24 hours. Next, the pegged lids were transferred to a fresh 96-well plate containing 180 mL of TSB and incubated overnight at 37 °C. MBEC values were determined as the lowest test concentration that resulted eradicated biofilm (i.e., wells displaying no turbidity in the final plate).
[0184] Pleased by the activity of the phosphonium compounds, the inventors sought to further investigate the antibacterial properties while comparing the effects of the heteroatom on activity. To do so, the inventors determined the minimum biofilm eradication concentration (MBEC) of bisQPC MT2P- 12, 12 and analogous bisQAC 12(2)12 using a pegged-lid microtiter plate assay. Both quaternary amphiphiles demonstrated an ability to eradicate established biofilm against three of the strains tested (Table 2).
[0185] Table 2 shows minimum Biofilm Eradication Concentrations of bisQPC MeP2P- 12,12 and bisQAC 12(2)12, as well as QAC standard DDAC.
[0186] Table 2
Figure imgf000052_0001
[0187] MBEC values for bisQAC 12(2)12 differed from a previous report, which highlights the effect that experimental method and growth conditions may have in antibacterial evaluations. While only minimal difference in potency was observed between the ammonium and phosphonium analogs, these results demonstrated that the novel cationic biocides provided in the present disclosure have superior biofilm eradication potency over alkyl commercially available QACs like didecyldimethylammonium chloride (DDAC).
[0188] Finally, the hemolysis (lysis20) assays for RP2P-n,n compounds correlated well with antimicrobial activity and alkyl chain length (n); no observed impact corresponded to the substituting group (R). The best balance between lysisio values and antibacterial activity was observed in the CyP2P-9,9, which displayed MIC values of 16 μM for most of the resistant bacterial strains and a modest lysis20 value of 125 pM. This represents an improvement in antimicrobial performance with diminished hemolysis over the commercial B AC control.
[0189] A novel series of alkyl based bisQPC compounds has been synthesized and evaluated for the ability to serve as disinfectants against a panel of bacteria, including those with previously reported AMR. The resulting compounds continue to demonstrate the effectiveness of bisQPCs as viable antibacterials, with improved bioactivity against multiple antibiotic-resistant bacterial strains over a commercial benchmark of BAC. The substituting groups of methyl, ethyl, cyclohexyl and phenyl appended to the phosphonium cations in the bisQPCs imparted only subtle differences in bioactivity across this RP2P-n,n structural motif. The largest determinant leading to low MIC values was the length of the amphiphilic tail, with 10-12 carbons continuing to serve as the optimum alkyl chain length. Comparison of the structural analogs of MT2P-12,12 and 12(2)12 showed highly analogous bioactivity and dynamic surface tension despite the change from a bisQPC to bisQAC in the respective compounds.
[0190] The experimental compounds shown above include two carbons in the linking group. However, the linking group is not limited to two carbons only. Based on the inventors’ research, the compounds with longer linking groups, for example, with 3-6 carbons or a cyclic group such as a cyclohexyl group are expected to have excellent antimicrobial performance. Examples of these compounds include, but are not limited to the following:
Figure imgf000053_0001
[0191] Each of these compounds comprises m X", for example, 2 Br", as counter ions.
When the counter ions are 2 Br, the compounds C22, C23, and C24 are coded as MT4P-10,10- Br, MT6P-10,10-Br,MTCyP-10,10-Br. The compounds C22, C23, and C24 are shown for illustration pinpose only. The methyl group can be replaced with other alkyl groups such as ethyl and cyclohexyl. The linking group L may have three or five carbons. The value of n can be in a range of from 6 to 12, for example. In addition to Br, the counterions can be any other type as described herein. [0192] Although the subject matter has been described in terms of exemplary embodiments, it is not limited thereto. Rather, the appended claims should be construed broadly, to include other variants and embodiments, which may be made by those skilled in the art.

Claims

What is claimed is:
1. An antimicrobial composition comprising a compound having the formula
Figure imgf000055_0001
wherein: n is an integer in a range of from 2 to 20,
L is a linking group being C1-10 alkyl or a cycloalkyl,
R1, R2, R3, or R4 each is a C1-12 alkyl or a cycloalkyl, unsubstituted or optional substituted with a functional group selected from the group consisting of -OH, -OR’, -NH2, -NHR’, -NR’2, - SH, -SR’, -O-C(O)R’, -C(O)OR’, -C(O)R’, -CF3, -OCF3, halogen, and any combination thereof;
R’ is H or a C1-4 alkyl;
X is selected from the group consisting of a halogen, a tosylate, a hydrocarbonate, a carbonate, a sulfate, and an acetate; and m is 1 or 2.
2. The antimicrobial composition of claim 1, wherein n is in a range of from 6 to 16.
3. The antimicrobial composition of claim 1, wherein n is in a range of from 10 to 12.
4. The antimicrobial composition of claim 1, wherein L is Ci-6 alkyl.
The antimicrobial composition of claim 1, wherein R1, R2, R3, or R4 each is a C1-6 alkyl or a cyclohexyl.
6. The antimicrobial composition of claim 1, wherein X is a halogen selected from F, Cl, Br, and I, and m is 2.
7. The antimicrobial composition of claim 1, wherein the compound is selected from the group consisting of:
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
wherein each compound comprises m X" as counter ions.
8. The antimicrobial composition of claim 1, wherein the antimicrobial composition is a disinfectant configured to be applied to a surface of an object in need thereof.
9. The antimicrobial composition of claim 1, wherein the antimicrobial composition is a pharmaceutical composition configured to be administrated to a subject in need thereof.
10. The antimicrobial composition of claim 9, wherein the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an active ingredient.
11. The antimicrobial composition of claim 9, wherein the pharmaceutical composition comprises an effective amount of the compound having the formula (I) as an additive for stabilizing the pharmaceutical composition.
12. A method of making the antimicrobial composition of claim 1 comprising preparing the compound having the formula (I).
13. The method of claim 12, further comprising mixing an effective amount of the compound having the formula (I) and a carrier.
14. A method of killing, preventing, or inhibiting microbial growth, comprising applying the antimicrobial composition of claim 1 to a surface of an object in need thereof.
15. A method of killing, preventing, or inhibiting microbial growth, comprising administrating the antimicrobial composition of claim 1 to a subject in need thereof.
16. A compound for antimicrobial use having the formula
Figure imgf000059_0001
wherein: n is an integer in a range of from 2 to 20,
L is a linking group being C1-10 alkyl or a cycloalkyl, R1, R2, R3, or R4 each is a C1-12 alkyl or a cycloalkyl, unsubstituted or optional substituted with a functional group selected from the group consisting of -OH, -OR’, -NH2, -NHR’, -NR’ 2, - SH, -SR’, -O-C(O)R’, -C(O)OR’, -C(O)R’, -CF3, -OCF3, halogen, and any combination thereof;
R’ is H or a C1-4 alkyl;
X is selected from the group consisting of a halogen, a tosylate, a hydrocarbonate, a carbonate, a sulfate, and an acetate; and m is 1 or 2.
17. The compound of claim 16, wherein n is in a range of from 6 to 16, and L is C1-6 alkyl.
18. The compound of claim 16, wherein R1, R2, R3, or R4 each is a C1-6 alkyl or a cyclohexyl.
19. The compound of claim 16, wherein X is a halogen selected from F, Cl, Br, and I, and m is 2.
20. The compound of claim 16, wherein the compound is selected from the group consisting of:
Figure imgf000060_0001
Figure imgf000061_0001
Figure imgf000062_0001
Figure imgf000063_0001
wherein each compound comprises m X" as counter ions.
PCT/US2023/079774 2022-11-15 2023-11-15 Phenyl-free biscationic quaternary phosphonium compounds as antimicrobial agents WO2024107810A1 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US202263425604P 2022-11-15 2022-11-15
US63/425,604 2022-11-15
US202263431801P 2022-12-12 2022-12-12
US63/431,801 2022-12-12

Publications (1)

Publication Number Publication Date
WO2024107810A1 true WO2024107810A1 (en) 2024-05-23

Family

ID=91085482

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2023/079774 WO2024107810A1 (en) 2022-11-15 2023-11-15 Phenyl-free biscationic quaternary phosphonium compounds as antimicrobial agents

Country Status (1)

Country Link
WO (1) WO2024107810A1 (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3364107A (en) * 1965-08-23 1968-01-16 American Cyanamid Co Controlling algae, bacteria, and fungi growth with alpha, omega-alkylenebis[triphenyl-phosphonium salt]
US20080275003A1 (en) * 2003-11-13 2008-11-06 The University Of Sydney Bis-Cationic Compounds and Use Thereof
CN102391660A (en) * 2011-09-26 2012-03-28 广东迪美生物技术有限公司 Green multifunctional composite aid for wood plastic composite material and preparation method thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3364107A (en) * 1965-08-23 1968-01-16 American Cyanamid Co Controlling algae, bacteria, and fungi growth with alpha, omega-alkylenebis[triphenyl-phosphonium salt]
US20080275003A1 (en) * 2003-11-13 2008-11-06 The University Of Sydney Bis-Cationic Compounds and Use Thereof
CN102391660A (en) * 2011-09-26 2012-03-28 广东迪美生物技术有限公司 Green multifunctional composite aid for wood plastic composite material and preparation method thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
SOMMERS KYLE J, MICHAUD MARINA E.; HOGUE CODY E.; SCHARNOW AMBER M.; AMOO LAUREN E.; PETERSEN ASHLEY A.; CARDEN ROBERT G.; MINBIOL: "Quaternary Phosphonium Compounds: An Examination of Non-Nitrogenous Cationic Amphiphiles That Evade Disinfectant Resistance", ACS INFECTIOUS DISEASES, AMERICAN CHEMICAL SOCIETY, US, vol. 8, no. 2, 11 February 2022 (2022-02-11), US , pages 387 - 397, XP093175104, ISSN: 2373-8227, DOI: 10.1021/acsinfecdis.1c00611 *
THIERER LAURA M, PETERSEN ASHLEY A.; MICHAUD MARINA E.; SANCHEZ CHRISTIAN A.; BRAYTON SAMANTHA R.; WUEST WILLIAM M.; MINBIOLE KEVI: "Atom Economical QPCs: Phenyl-Free Biscationic Quaternary Phosphonium Compounds as Potent Disinfectants", ACS INFECTIOUS DISEASES, AMERICAN CHEMICAL SOCIETY, US, vol. 9, no. 3, 10 March 2023 (2023-03-10), US , pages 609 - 616, XP093175106, ISSN: 2373-8227, DOI: 10.1021/acsinfecdis.2c00575 *

Similar Documents

Publication Publication Date Title
US20230365491A1 (en) Method for inhibiting or disrupting biofilm formation, or reducing biofilm
AU2017201670B2 (en) A composition comprising an antibiotic and a dispersant or an anti-adhesive agent
JP4994033B2 (en) N, N-dihalogenated amino acids and derivatives
US8748617B2 (en) Amide compound or salt thereof, and biofilm inhibitor, biofilm remover and disinfectant containing the same
CZ301637B6 (en) Thixotropic hydrogel disinfectant
KR20070098947A (en) N-halogenated amino acids and n,n-dihalogenated amino acids in combination with hypohalous acids
JP2017218457A (en) Derivatives of xanthone compounds
EP3638251A1 (en) Bisphosphocin gel formulations and uses thereof
US8552208B2 (en) Analogs of tetramic acid
US20160106689A1 (en) Use
WO2016046523A1 (en) Use of cystamine for treating bacterial and fungal infections
EP2077260B1 (en) Amide compound or salt thereof, and biofilm formation inhibitor, biofilm remover and bactericide each using the amide compound or salt thereof
WO2024107810A1 (en) Phenyl-free biscationic quaternary phosphonium compounds as antimicrobial agents
US20230312610A1 (en) Anti-Microbial Compositions
WO2024129836A1 (en) Biscationic quaternary phosphonium compounds as soft antimicrobial agents
EP4387628A1 (en) Quaternary phosphonium compounds and uses thereof
KR20080086502A (en) Diphenyl urea derivatives
CA3106105A1 (en) Methods of diagnosing and treating infected implants