Patent Application
20180221309
Bloodstream infections affect a huge patient population in the United States, with more than 250,000 cases reported each year. Patients with indwelling medical devices, such as central venous catheters (CVCs), are most at risk for these infections. Frequently, various microorganisms from the skin of the patient, or respective healthcare professional, can gain access through the catheter wound as a result of non-sterile conditions. Of these resulting bloodstream infections, Candida species account for 9% of all bloodstream infections and are associated with ˜40% mortality rate. The most commonly isolated fungal pathogen from bloodstream infections is Candida albicans, but the prevalence of other species, such as C. parapsdosis, C. glabrata, and C. tropicalis, is increasing.
Candida spp. pathogens possess an outer cell wall that is an important determinant of pathogenicity. The cell wall is primarily composed of carbohydrates and. structurally is separated into two layers. The outer layer is composed mostly of N-linked glycans and mannoproteins and the inner layer is composed of, β-glucan and chitin. The complexity of the cell wall contributes to various pathogenic factors including adherence of the fungus and establishment of cross-talk with the host known as “glycan code.” Cell wall components are also found in the extracellular matrix secreted by Candida spp. biofilms, which can contaminate the synthetic material surfaces of indwelling medical devices. Candida spp. biofilm production of polysaccharides, such as β-glucan, contributes to the decreased susceptibility of biofilms to antifungal drugs by sequestering antifungal drugs.
Various antimicrobial impregnation approaches have been devised to prevent catheter infections. Catheter materials coated with chlorhexidine-silver sulfadiazine and minocycline/rifampin have shown trends in reduced infection rates, but their clinical effectiveness remains questionable. Other treatments, including the use of silver-impregnated subcutaneous collagen cuffs, have also failed to be effective in recent trials. CVC contamination generally requires removal and replacement of the device in addition to a prolonged course of antifungal drug therapy, which raises concerns regarding drug toxicity and development of antifungal resistance. Antifungal chemotherapy is also problematic, with increasing prevalence of resistance to azole and echinocandin drugs as well as well-known nephrotoxicity of amphotericin B. Due to the high morbidity and mortality rate of catheter-related Candida spp. bloodstream infections, strategies for preventing medical device contamination by fungal pathogens remains a top priority for infection control.
In various embodiments, a method of treating a fungal infection includes contacting a fungus including a β-glucan that is at least partially masked from immune system detection with a therapeutically effective amount of a compound that at least partially unmasks the β-glucan to increase immunogenicity of the fungus.
In various embodiments, a method of treating a fungal infection includes contacting a fungus including a β-glucan that is at least partially masked from immune system surveillance with a therapeutically effective amount of a compound that at least partially unmasks the β-glucan to increase immunogenicity of the fungus, wherein the compound includes a unit having the structure:
The variable A1 is chosen from a bond,
The variable A2 is chosen from a bond,
The variable R1 is chosen from —H and C1, wherein the compound includes at least one C1, at each occurrence, C1 is independently chosen from -L-D and —O-L-D, at each occurrence, L is independently (C1-C10)hydrocarbylene, at each occurrence, D is independently chosen from —N((C1-C5)alkyl)3 and a (C1-C10)alkyl-substituted cationic nitrogen-containing (C1-C5)heterocycle, and n is about 1 to about 100,000.
In various embodiments, an antifungal compound includes a unit having the structure:
The variable A2 is chosen from a bond,
The variable A2 is chosen from a bond,
The variable R1 is chosen from —H and C1, wherein the compound includes at least one C1, at each occurrence, C1 is independently chosen from -L-D and —O-L-D, at each occurrence, L is independently (C1-C10)hydrocarbylene, at each occurrence, D is independently chosen from —N((C1-C5)alkyl)3 and a (C1-C10)alkyl-substituted cationic nitrogen-containing (C1-C5)heterocycle, and n is about 1 to about 100,000, and wherein contact between a therapeutically effective amount of the compound and a fungus including β-glucan that is at least partially masked from immune system detection is effective to at least partially unmask the β-glucan thereby increasing immunogenicity of the fungus.
The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments of the present invention.
Reference will now be made in detail to certain embodiments of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.
Throughout this document, values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.
In this document, the terms “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. The statement “at least one of A and B” has the same meaning as “A, B, or A and B.” In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting; information that is relevant to a section heading may occur within or outside of that particular section. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.
In the methods described herein, the acts can be carried out in any order without departing from the principles of the invention, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.
The term “about” as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or of a stated limit of a range, and includes the exact stated value or range.
The term “substantially” as used herein refers to a majority of, or mostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%.
The term “organic group” as used herein refers to any carbon-containing functional group. Examples can include an oxygen-containing group such as an alkoxy group, aryloxy group, aralkyloxy group, oxo(carbonyl) group; a carboxyl group including a carboxylic acid, carboxylate, and a carboxylate ester; a sulfur-containing group such as an alkyl and aryl sulfide group; and other heteroatom-containing groups. Non-limiting examples of organic groups include OR, OOR, OC(O)N(R)2, CN, CF3, OCF3, R, C(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)0-2N(R)C(O)R, (CH2)0-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(═NH)N(R)2, C(O)N(OR)R, C(═NOR)R, and substituted or unsubstituted (C1-C100)hydrocarbyl, wherein R can be hydrogen (in examples that include other carbon atoms) or a carbon-based moiety, and wherein the carbon-based moiety can be substituted or unsubstituted.
The term “substituted” as used herein in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term “functional group” or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CF3, OCF3, R, O, (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)0-2N(R)C(O)R, (CH2)0-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(═NH)N(R)2, C(O)N(OR)R, and C(═NOR)R, wherein R can be hydrogen or a carbon-based moiety; for example, R can be hydrogen, (C1-C100)hydrocarbyl, alkyl. acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl; or wherein two R groups bonded to a nitrogen atom or to adjacent nitrogen atoms can together with the nitrogen atom or atoms form a heterocyclyl.
The term “alkyl” as used herein refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. As used herein, the term “alkyl” encompasses n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.
The term “alkenyl” as used herein refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, —CH═CH(CH3), —CH═C(CH3)2, —C(CH3)═CH2, —C(CH3)═CH(CH3), —C(CH2CH3)═CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.
The term “aryl” as used herein refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring. Thus aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, inda.cenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a. phenyl group substituted at any one or more of 2-, 3-, 4--, 5-, or 6-positions of the phenyl ring, or a naphthyl group substituted at any one or more of 2- to 8-positions thereof.
The term “heterocyclyl” as used herein refers to aromatic and non-aromatic ring compounds containing three or more ring members, of which one or more is a heteroatom such as, but not limited to, N, O, and S.
The term “heteroaryl” as used herein refers to aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members. A heteroaryl group is a variety of a heterocyclyl group that possesses an aromatic electronic structure.
The term “amine” as used herein refers to primary, secondary, and tertiary amines having, e.g., the formula N(group)3 wherein each group can independently be H or non-H, such as alkyl, aryl, and the like. Amines include but are not limited to R—NH2, for example, alkylamines, arylamines, alkylarylamines; RAH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, heterocyclylamines and the like; and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like. The term “amine” also includes ammonium ions as used herein.
The term “amino group” as used herein refers to a substituent of the form —NH2, —NHR, —NR2, —NR3′, wherein each R is independently selected, and protonated forms of each, except for —NR3′, which cannot be protonated. Accordingly, any compound substituted with an amino group can be viewed as an amine. An “amino group” within the meaning herein can be a primary, secondary, tertiary, or quaternary amino group. An “alkylamino” group includes a monoalkylamino, dialkylamino, and trialkylamino group.
The terms “halo,” “halogen,” or “halide” group, as used herein, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.
The term “hydrocarbon” or “hydrocarbyl” as used herein refers to a molecule or functional group, respectively, that includes carbon and hydrogen atoms. The term can also refer to a molecule or functional group that normally includes both carbon and hydrogen atoms but wherein all the hydrogen atoms are substituted with other functional groups.
As used herein, the term “hydrocarbyl” refers to a functional group derived from a straight chain, branched, or cyclic hydrocarbon, and can be alkyl, alkenyl, alkynyl, aryl, cycloalkyl, acyl, or any combination thereof. Hydrocarbyl groups can be shown as (Ca-Cb)hydrocarbyl, wherein a and b are integers and mean having any of a to b nurriber of carbon atoms. For example, (C1-C4)hydrocarbyl means the hydrocarbyl group can be methyl (C1), ethyl (C2), propyl (C3), or butyl (C4), and (C0-Cb)hydrocarbyl means in certain embodiments there is no hydrocarbyl group.
The term “number-average molecular weight” (Mn) as used herein refers to the ordinary arithmetic mean of the molecular weight of individual molecules in a sample. It is defined as the total weight of all molecules in a sample divided by the total number of molecules in the sample. Experimentally, Mn is determined by analyzing a sample divided into molecular weight fractions of species i having ni molecules of molecular weight Mi through the formula Mn=ΣMini/Σni. The Mn can be measured by a variety of well-known methods including gel permeation chromatography, spectroscopic end group analysis, and osmometry. If unspecified, molecular weights of polymers given herein are number-average molecular weights.
The term “weight-average molecular weight” as used herein refers to Mw, which is equal to ΣMi2ni/ΣMini is the number of molecules of molecular weight Mi. In various examples, the weight-average molecular weight can be determined using light scattering, small angle neutron scattering, X-ray scattering, and sedimentation velocity.
The term “oligomer” as used herein refers to a molecule having an intermediate relative molecular mass, the structure of which essentially includes a small plurality of units derived, actually or conceptually, from molecules of lower relative molecular mass. A molecule having an intermediate relative mass can be a molecule that has properties that vary with the removal of one or a few of the units. The variation in the properties that results from the removal of the one of more units can be a significant variation.
The term “solvent” as used herein refers to a liquid that can dissolve a solid, liquid, or gas. Non-limiting examples of solvents are silicones, organic compounds, water, alcohols, ionic liquids, and supercritical fluids.
Herein, when it is designated that a variable in the structure can be “a bond,” the variable can represent a direct bond between the two groups shown as linked to that variable, such as a single bond.
As used herein, the term “polymer” refers to a molecule having at least one repeating unit and can include copolymers.
In various embodiments, salts having a positively charged counterion can include any suitable positively charged counterion. For example, the counterion can be ammonium(NH4+), or an alkali metal such as sodium (Na+), potassium (K+), or lithium (Li+). In some embodiments, the counterion can have a positive charge greater than+1, which can in some embodiments complex to multiple ionized groups, such as Zn2+, Al3+, or alkaline earth metals such as Ca2+ or Mg2+.
In various embodiments, salts having a negatively charged counterion can include any suitable negatively charged counterion. For example, the counterion can be a halide, such as fluoride, chloride, iodide, or bromide. In other examples, the counterion can be nitrate, hydrogen sulfate, dihydrogen phosphate, bicarbonate, nitrite, perchlorate, iodate, chlorate, bromate, chlorite, hypochlorite, hypobrornite, cyanide, amide, cyanate, hydroxide, permanganate. The counterion can be a conjugate base of any carboxylic acid, such as acetate or formate. In some embodiments, a counterion can have a negative charge greater than −1, which can in some embodiments complex to multiple ionized groups, such as oxide, sulfide, nitride, arsenate, phosphate, arsenite, hydrogen phosphate, sulfate, thiosulfate, sulfite, carbonate, chromate, dichromate, peroxide, or oxalate.
In various embodiments, the polymers described herein can terminate in any suitable way. The polymers can terminate with an end group that is independently chosen from a suitable polymerization initiator, —H, —OH, a substituted or unsubstituted (C1-C20)hydrocarbyl (e.g., (C1-C10)alkyl or (C6-C20)aryl) interrupted with 0, 1, 2, or 3 groups independently chosen from —O—, substituted or unsubstituted —NH—, and —S—, a poly(substituted or unsubstituted C20)hydrocarbyloxy), and a poly(substituted or unsubstituted (C1-C20)hydrocarbylamino).
Candida cell walls are built on a scaffold of β-(1,3;1,6)-glucan fibrils. β-glucan is highly immunogenic due to its recognition by Dectin-1 and beta2 integrins, leading to phagocytosis and inflammatory activation of innate immunocytes. C. albicans efficiently masks glucan from innate immune surveillance, which helps it to evade host defense. Conditions that can unmask glucan can increase Dectin-1 dependent responses (i.e., phagocytosis) to Candida yeasts by leukocytes. In various embodiments, the compounds described herein can therapeutically lead to glucan unmasking via chemical species that will increase immunogenicity of Candida species pathogens and provoke stronger innate immune response to fungal infection. In various embodiments, the methods described herein can induce glucan unmasking via ROS-mediated cell wall damage using phenylene ethynylene (PE) antimicrobials using small molecule drug compounds. Achieving therapeutic glucan unmasking can have clinical applications in care of all types of wounds and in preventing and/or treating microbial contamination of medical devices of many types.
In various embodiments, the methods described herein can treat Candidiasis and other fungal infections by changing the immunogenicity of the cell wall surface in Candida species. The cell wall can be an excellent target for therapeutic intervention in Candidiasis and other fungal infections because it has no human counterpart, so interventions impacting its synthesis, remodeling or repair are less likely to be confounded by off-target side effects. Also, relieving immune evasion via therapeutic glucan unmasking can bring existing host defense mechanisms maximally to bear on the eradication of the pathogen. In various embodiments, the methods described herein when used in combination with existing antimycotic drugs can increase the effectiveness and useful lifetime of first line azole and echinocandin drugs in the face of intrinsic and acquired resistance.
Therapeutic glucan unmasking can be important in the treatment of fungal infectious disease due to the potential breadth of its application to many common fungal pathogens. For example, emerging non-albicans Candida species pathogens, Aspergillus and Histoplasma species all engage in glucan masking. Moreover, β-glucan is ubiquitous in the cell walls of fungi, so it is a target of very broad significance to mycoses. In various embodiments, the methods described herein can be used to treat infections due to organisms that engage in glucan masking.
In various embodiments, the methods described herein provide therapeutic glucan unmasking in Candida species using light-activated antimicrobial agents. Phenylethynylenes (PEs) are antimicrobial agents with a high singlet oxygen yield upon light exposure. As shown in the Examples herein, in various embodiments, a polycationic PE (PPE-DABCO) can bind Candida cell walls, effecting light-activated glucan unmasking, and leading to increased phagocytosis. Other PE compounds can also be used to induce glucan exposure for the purpose of increasing the immunogenicity of the pathogen surface and elevating the host's immune response to the pathogen. In particular, PE compounds that include a thiophene moiety can induce glucan exposure. The method can increase the immunogenicity of any fungal pathogen that contains β-glucan in its cell wall, which is a common feature in fungal pathogens. Some major human fungal pathogen genera that can be susceptible to therapeutic glucan unmasking can include Candida, Histoplasma, and Aspergillus. PE-based glucan unmasking methods can have particular application to external medical settings such as wound care. In various embodiments, the methods described herein can be used to treat topical infections, for prevention or treatment of wound infections, for treatment applied to wound dressings, for treatments applied to treat or modify surfaces of medical devices that can become microbially or fungally contaminated , and to prevent or treat medical device infection.
In various embodiments, the methods described herein provide a general technique of using small molecule drug compounds to induce therapeutic glucan unmasking in C. albicans. Fungal enzymes synthesize and remodel cell wall structure, maintaining glucan masking. These enzymes are potential drug targets to induce therapeutic glucan unmasking. Drug design and/or drug screening approaches can be used to identify small molecule compounds that induce glucan unmasking in fungal pathogens such as C. albicans yeast.
In various embodiments, a method of treating a fungal infection is provided. The method includes contacting a fungus including a β-glucan that is at least partially masked from immune system detection with a therapeutically effective amount of a compound that at least partially unmasks the β-glucan to increase immunogenicity of the fungus. The method can include contacting the fungus with one or more compounds. The compound can be any suitable compound that can be used to carry out the method as described herein, and can be any compound described herein. In various embodiments, the compound is a polycationic conjugated aromatic system.
The contacting between the fungus and the compound can be any suitable contacting. The contacting can be contacting between the fungus and a solution including the compound. For example, the contacting can be contacting between a fungus in a catheter and a solution including the compound that has been injected into the catheter. The contacting can be contacting between the fungus and the surface of a substrate including the compound.
The therapeutically effective amount of the compound can be any suitable concentration, such as a concentration of about 0.001 mg/L to about 1000 g/L, or about 0.001 g/L to about 100 g/L, or about 0.001 mg/L or less, or less than, equal to, or greater than about 0.01 mg/L, 0.1 mg/L, 1 mg/L, 0.01 g/L, 0.1 g/L, 1 g/L, 10 g/L, 100 g/L, or about 1000 g/L, or more.
The β-glucan can be any suitable β-glucan, and can include β-(1,3)-glucan, β-(1,6)-glucan, β-(1,3;1,6)-glucan, or a combination thereof. The β-glucan can include β-(1,3;1,6)-glucan, such as β-(1,3;1,6)-fibrils, such as in fungal cell walls.
The fungus can be any one or more fungi. The fungus can be a Candida species fungus, an Aspergillus species fungus, a Histoplasma species fungus, a Blastomyces species fungus, a Coccidioides species fungus, a Cryptococcus species fungus, a Fusarium species fungus, a Sporothrix species fungus, a Rhizopus species fungus, a Mucor species fungus, a Rhizomucor species fungus, a Cunninghamella species fungus, a Absidia species fungus, a Saksenaea species fungus, a Apophysomyces species fungus, a Paracoccidioides species fungus, a Trichophyton species fungus, a Microsporum species fungus, a Epidermophyton species fungus, or a Malassezia species fungus. The fungus can be Candida albicans, Candida glabrata, Candida parapsilosis, or a combination thereof. The fungus can include a biofilm (e.g., the fungus can be part of a biofilm). The fungus can be in or on an indwelling medical device (e.g., an implanted or inserted medical device, such as a medical implant or a catheter). The fungus can be in or on a catheter. The fungus can be in or on any medical device material. The fungus can be in or on any human tissue as a commensal or pathogenic organism.
The compound can include a unit having the structure:
The variable A1 can be chosen from a bond,
The variable A2 can be chosen from a bond,
The variable R1 can be chosen from —H and C1. The compound can include at least one C1 (e.g., at least one cationic group, or at least two cationic groups). At each occurrence, C1 can be independently chosen from -L-D and —O-L-D. At each occurrence, L can be independently (C1-C10)hydrocarbylene. At each occurrence, D can be independently chosen from ‘N((C1-C5)alkyl)3 and a (C1-C10)alkyl-substituted cationic nitrogen-containing (C1-C5)heterocycle. The variable n can be about 1 to about 100,000, about 1 to about 20, about I to about 10, or about 1, or less than, equal to, or greater than about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or about 20 or more.
The compound can have the structure:
At each occurrence, the variable T can be independently chosen from —H, C(O)—O—(C1-C10)alkyl, -phenyl, and —R1.
The variable L can be independently chosen from methylene, ethylene, propylene, butylene, pentylene, and heptylene.
The variable A1 can be chosen from a bond,
At each occurrence, the variable D can e independently chosen from —N(CH3)3,
At each occurrence, the variable D can be independently chosen from —N(CH3)3,
The variable C1 can be chosen from:
At each occurrence, the variable T can be independently chosen from —H, C(O)—O—ethyl, -phenyl, and —R1.
The compound can be:
The compound can be a polymer (e.g., a polymer or copolymer) including the repeating unit:
The compound can be a polymer (e.g., a homopolymer or copolymer) including the repeating unit:
The compound can be a polymer (e.g., a homopolymer or copolymer) including the repeating group:
The compound can be a polymer (e.g., a homopolymer or copolymer) including the repeating group:
The compound can be a polymer (e.g., a homopolymer or copolymer) including the repeating group:
The compound can be:
The compound can be:
The compound can be:
The compound can be:
The compound can be:
The compound can be:
The compound can be:
The compound can be:
The compound can be:
The compound can be:
In various embodiments, the method described herein can prevent or reduce a fungal infection on a substrate. The method can include treating the substrate with a therapeutically effective amount of a compound so that contact between the treated surface and a fungus including β-glucan that is at least partially masked from immune system detection is effective to at least partially unmask the β-glucan thereby increasing immunogenicity of the fungus and preventing or reducing fungal infection on the substrate from the fungus. At the time of treatment, the substrate can be substantially free of the fungus, or the substrate can include the fungus. The compound can be any suitable compound, such as any compound described herein.
The method can include filling or coating a medical device, such as a catheter, with a solution including the compound.
In various embodiments, the method of preventing or reducing a fungal infection on or in a device. The device can be any device, such as a medical device, such as a catheter. The method can include treating the device with a therapeutically effective amount of a compound so that contact between the compound and a fungus including β-glucan that is at least partially masked from immune system detection is effective to at least partially unmask the β-glucan thereby increasing immunogenicity of the fungus and preventing or reducing fungal infection on or in the device from the fungus. The compound can be any suitable compound, such as any compound described herein.
The method can include filling or coating a medical device, such as a catheter, with a solution including the compound.
In various embodiments, any one of the compounds described herein is an antifungal compound. Contact between a fungus including a β-glucan that is at least partially masked from immune system detection and a therapeutically effective amount of the antifungal compound can at least partially unmasks the β-glucan to increase immunogenicity of the fungus. The compound can be any suitable compound that can be used to perform the methods described herein. The compound can be any suitable compound described herein.
Various embodiments provide a device (e.g., a medical device) or a substrate that includes one or more of the compounds, wherein the device experiences less or no fungal infections, as compared to a device or substrate free of the one or more compounds.
The antifungal compound can include a unit having the structure:
The variable A1 can be chosen from a bond,
The variable A2 can be chosen from a bond,
The variable R1 can be chosen from —H and C1. The compound can include at least one C1. At each occurrence, C1 can be independently chosen from -L-D and —O-L-D. At each occurrence, L can be independently (C1-C10)hydrocarbylene. At each occurrence, D can be independently chosen from —N((C1-C5)alkyl)3 and a (C1-C10)alkyl-substituted cationic nitrogen-containing C5)heterocycle. The variable n can be about 1 to about 100,000. Contact between the compound and a fungus including β-glucan that is at least partially masked from immune system detection is effective to at least partially unmask the β-glucan thereby increasing immunogenicity of the fungus.
The compound can have the structure:
The variable R2 can be chosen from a bond and
The variable R3 can be chosen from a bond and
At each occurrence, T can he independently chosen from —H, C(O)—O—(C1-C10)alkyl, -phenyl, and —R1.
The compound can he ori ude any of the compounds shown in Table 1.
Various embodiments of the present invention can be better understood by reference to the following Examples which are offered by way of illustration. The present invention is not limited to the Examples given herein. In the Examples herein, references to “glucan” refer to β-glucan.
The antimicrobial effectiveness of two phenylene ethynylene (PE) compounds against Candida species. A subset of conjugated polyelectrolytes, phenylene ethynylenes have shown promising biocidal activity against Gram-positive and -negative bacterial pathogens, as well as the environmental yeast, Saccharomyces cerevisiae. The chemical structure of these compounds renders them capable of inducing broad-spectrum cell damage. The phenylene ethynylenes studied include compounds with alternating phenyl and acetylenic groups with appended cationic groups (Scheme 1). The interaction of the cationic quaternary ammonium groups with net-anionic membranes and cell walls facilitates interactions with cells, leading to extensive leakage of cell contents. In addition, when PE compounds such as these are irradiated by the appropriate wavelength of light, the backbone produces reactive oxygen species (ROS) that induce rapid cell death.
The oligomeric and polymeric molecular size of PE compounds can play a role in their mechanisms of killing. Additionally, the antimicrobial activity of these compounds can dependent on various factors that include molecular conformation, size, functional groups, and, the membrane composition of the target pathogen. After treatment with the compounds, the viability of Candida spp. was monitored using flow cytometry.
Candida albicans (ATCC, #MYA-2876), C. parapsilosis (ATCC, #22019), and C. glabrata (ATCC, #2001) were grown from glycerol stocks, stored at −80° C. Said stock was transferred to 5 mL filtered yeast extract-peptone-dextrose (YPD) medium (Becton Dickinson), and grown for 16 h at 30° C., with a shaking speed of 300 RPM. These growth conditions yielded yeast at the mid exponential phase.
Following a 10 minute centrifugation at 4,400 RPM, the supernatant was replaced with sterile phosphate-buffered saline (PBS), and subsequently vortexed. This washing step is repeated a second time to mitigate cell debris. Cell concentration was then determined using a disposable hemocytometer (INCYTO C-Chip; Fisher Scientific).
Patient specimens (peripheral blood or catheter tips) were processed by Tricore Reference Laboratories (Albuquerque, N. Mex.) and identified as C. albicans using a Bruker Biotyper MALDI-TOF system (MS ID score>2.0). Clonal isolates so identified were subcultured on Sabouraud agar slants and provided to the investigators as unique strains. Isolate strains were provided in completely deidentified form according to procedures approved by the University of New Mexico School of Medicine Human Research Protections Office. For biocidal assays, clinical isolates were grown in YPD broth as described above.
Biocidal experiments were carried out in either translucent or opaque 1.5 mL microcentrifuge tubes, at cell concentrations of 5×106/mL. EO-OPE-DABCO and PPE-DABCO stocks were prepared in sterile deionized water (18.2 MΩ·cm at 25° C.), and contained 0.47% dimethyl sulfoxide (by volume) to improve solubility and minimize aggregate formation. Negative controls contained equal amounts of dimethyl sulfoxide.
Samples were exposed to controlled amounts of light using a 14-lamp photoreactor (LZC-4V; Luzchem Research; Ottawa, Ontario). A rotating carousel ensured that all samples receive equivalent levels of light exposure; ventilation kept the photoreactor below 30° C. EO-OPE-DABCO absorbs in the ultraviolet region, warranting the use of UVA lamps (350 nm emission peak; 4.46±2.41 mW/cm2) to optimize singlet oxygen yields. Conversely, 420 nm blue-light lamps (6.62±2.93 mW/cm2) were used in PPE-DABCO tests; unlike its oligomeric counterpart, polymeric PPE-DABCO absorbs in the near-visible range. Power density output was measured at the peak excitation wavelength for both lighting configurations. Data shown is an aggregation of two independent replicate experiments.
Samples were then stained with 5 μM membrane-permeable SYTO 9 and 1 μM membrane-impermeable TO-PRO-3, both of which are nucleic acid stains. After 30 min, samples were evaluated by flow cytometry (FACSCalibur; Becton Dickinson). At least 10,000 events were evaluated in each trial. A heat-killed sample (70° C. for 30 min) was used to identify the fluorescence characteristics of dead cells.
15 min dark-activity assays were carried out in a somewhat different manner. Samples were prepared and stained with SYTO 9 and TO-PRO-3, albeit in the absence of any biocide. After a 30 min staining duration, EO-OPE-DABCO was added (10 μg/mL final concentration); the sample was then vortexed and analyzed by flow cytometry. Every minute, viability data was collected (again, 10,000 events/sample), for a total of 15 minutes. EO-OPE-DABCO was added one sample at a time, so that, in each case, flow cytometry readings could begin within 1 min of the biocide's introduction.
Stocks of S. cerevisiae β-(1,3)-glucan (high, medium or low MW; gift of Biothera, Eagan, Minn.), PPE-DABCO, and EO-OPE-DABCO were mixed with 10 mM pH 7.4 phosphate buffer to a final concentration of 2 μg/mL in PPE-DABCO or EO-OPE-DABCO and 100 μg/mL in glucan. 200 μL solutions were transferred to a 160 μL nominal volume fused quartz fluorimetry cuvette, and read on a PTI QuantaMax 40 steady-state fluorescence spectrophotometer (HORIBA Scientific, Edison, N.J.) with PMT detection. Emission spectra were obtained using an excitation wavelength of 350 nm for EO-OPE-DABCO and 420 nm for PPE-DABCO, and excitation spectra were obtained with the corresponding maximum emission wavelength.
C. albicans yeast cells were treated in a similar manner to that of the previously described biocidal experiments. In effort to maintain a consistent degree of cell death across samples, OPE-DABCO exposure in UVA light was limited to just 10 min. A thermal positive control was also implemented, which entailed heating samples to 100° C. for 30 min. Following the appropriate treatment and removal from the photoreactor, samples were blocked with 1% (w/v) bovine serum albumin (BSA) for 30 min at room temperature. The samples were then treated with a primary antibody, anti β-glucan IgG, at a final concentration of 10 μg/mL, for an additional 30 min. Negative controls contained 10 μg/mL isotype-matched murine IgG in place of anti β-glucan IgG. A secondary antibody with Alexa Fluor 647 dye was then added (1 ug/ml in PBS1% BSA), along with 5 μM SYTO 9 were simultaneously added and allowed to stain cells for 30 minutes at 25° C. prior to analysis by flow cytometry. Data shown is an aggregation of two independent replicate experiments.
HEK-293 cells (ATCC, #CRL-1573) were cultured in DMEM supplemented with 10% CS, 1% Penicillin/Streptomycin, 2mM L-glutamine, and 1 mM sodium pyruvate at 37° C., 5% CO2. Cells were then plated in 6 well plate at 1×105 cell per well. mApple-human Dectin1A-C-10 (addgene, #54883) was transfected into cells by following standard protocols using Fugene 6 (Promega, #E2691). Cell cultures were used for further experimentation at 24 h post-transfection with growth in normal medium, as described above.
C. albicans yeast cells were subjected to the same treatment conditions as in the aforementioned β-glucan exposure study, before being spun down and washed in PBS. Following the last wash step the C. albicans were stained with 7.5 μM of SYTO 9 (Invitrogen, #S-34854) and 7.5 μM of CypHer5E NHS-ester (GE Healthcare, #PA15401) for one hour at 25° C. After staining the C. albicans were added to live, Dectin-1A-C-10 transfected HEK-293 cells for one hour. Next ice cold PBS was used to lift the HEK-293 cells off of the plate. Either controls with C. albicans or HEK-293 alone or the above samples with a mixture of C. albicans and HEK-293 cells were analyzed using an LSR Fortessa flow cytometer (Becton Dickinson) and FlowJo software (FlowJo, Ashland, Oreg.). At least 10,000 side scatter (SSC)-positive events are evaluated in each trial. CypHer 5, SYTO 9, and mApple fluorescence was observed at emission wavelengths of 660 nm (670/14), 525 nm (505 LP, 530/30), and 578 nm (582/15), respectively. Data shown is an aggregation of two independent replicate experiments.
A series of biocidal studies were carried out to gain insight to the light-activated effects of EO-OPE-DABCO and PPE-DABCO on Candida species pathogens. Phenylene ethynylenes are unique in that their mechanism of action differs, depending on the presence of light; in particular, light intensity, emission wavelength, and duration. In the studies described herein, duration of light exposure was the primary variable being studied. Light intensity was kept constant using a photoreactor with 14 interchangeable lamps. Lamps were chosen to have an emission wavelength overlapping the excitation spectrum of the phenylene ethynylene being used. 350 nm--centered INA lamps were implemented for EO-OPE-DABCO testing, while 420 nm-centered lamps were used in PPE-DABCO tests. With light intensity and spectrum being held constant for a given phenylene ethynylene, the effect of light exposure duration was investigated to discern C. albicans' susceptibility to phenylene ethynylenes in the light vs. dark. Even though all samples were exposed to one of the two compounds for a total of 60 min, the duration of light exposure was varied by 4 min intervals and the balance of 60 min exposure was in the dark.
Interactions between both PEs with soluble β-(1,3)-glucan extracted from Saccharomyces cerevisiae yeast cell walls were evaluated. The structure of S. cerevisiae and C. albicans β(1.3)-glucan is similar, and this polysaccharide is an important part of Candida drug resistance and pathogenicity, amounting to 40% of the cell wall. Size fractionated β-glucan (low MW=11 kDa, medium MW=150 kDa, high MW=450 kDa) were tested (
Although more profound in the case of PPE-DABCO, enhanced emission of both PEs upon the introduction of the high molecular weight β-glucan was observed, which is indicative of complexation. In addition, a small degree of red-shifting was observed, suggesting that rotation of the conjugated regions of the PEs are restricted due to complexation with soluble β-glucan. Lateral hydrogen bonding between β-glucan polymers can facilitate PE/β-glucan complexation, analogous to their role in stabilizing lateral interactions of individual β-glucan polymers in aqueous solution. These observed photophysical changes were more dramatic with PPE-DABCO than EO-OPE-DABCO, suggesting that increased complexation of PPE-DABCO can be due to its size, which is substantial relative to that of an oligomenc molecule. PPE-DABCO has numerous sites where weak interactions with glucan polymers may form; furthermore, extensive valency of laterally-aggregated β-glucan would make this interaction very strong. EO-OPE-DABCO is far smaller than PPE-DABCO, and therefore exhibits a lower-avidity interaction with β-glucan. Without being bound by theory, these results can help explain PPE's inability to kill C. albicans in the dark. Exhibiting a strong propensity to interact and associate with β-glucan, it is likely that PPE-DABCO is limited in its ability to fully penetrate the cell wall and much of the compound is sequestered on β-glucan in the cell wall. Given the limited radius of destruction of singlet oxygen and the density of organic material in the cell wall capable of quenching singlet oxygen, this association may be limiting the depth of cell wall permeation of PPE-DABCO and its capacity to directly perturb the yeast's plasma membrane, relative to EO-OPE-DABCO. The oligomer, on the other hand, appears far less likely to interact with β-glucan, which can allow it to permeate the fungal cell wall more readily and better access the yeast's plasma membrane.
These results shed light onto the mechanisms by which EO-OPE-DABCO effectively kills C. albicans yeast cells (
Using a modified biocidal assay, six C. albicans clinical isolates were surveyed for their susceptibility to 10 μg/mL EO-OPE-DABCO in the dark. In this instance, the cells were stained with SYTO 9 and TO-PRO-3 before the introduction of EO-OPE-DABCO. Taking a flow cytometry dual--fluorescent measurement of 10,000 events every minute allowed for real-time reporting of EO-OPE-DABCO-induced membrane perturbation. The susceptibility of clinical isolates was gauged relative to that of C. albicans SC5314, as shown in
Variability of susceptibility to EO-OPE-DABCO amongst clinical isolates of one species (C. albicans) suggested that non-albicans Candida species pathogens might also exhibit variable sensitivity to this biocide. The aforementioned 15 min flow cytometry assay was used to determine if EO-OPE-DABCO was more or less effective against C. parapsilosis and C. glabrata relative to C. albicans SC5314.
β-glucan is highly immunogenic upon recognition by the innate immunoreceptors Dectin-1 or Mac-1. Several prominent genera of fungal pathogens, including Candida, are known to employ an innate immune evasive strategy of masking glucan to restrict its exposure on the cell wall surface. Without being bound by theory, it is believed that PE antimicrobials bound to cell wall constituents and exposed to light can generate singlet oxygen, leading to local cell wall damage, unmasking β-glucan and increasing immunogenicity. Using an anti-β-(1,3)-glucan primary antibody in tandem with a secondary fluorescently-labeled antibody allowed for the comparison of glucan exposure following treatment conditions: PE in the dark, PE in the light, and a 60 min light negative control. C. albicans yeast treated with EO-OPE-DABCO in the dark, using conditions associated with high biocidal activity (
Given that PPE-DABCO can increase β-glucan exposure on C. albicans yeast, a test of whether the unmasking achieved by this treatment resulted in greater recognition of yeasts through the β-glucan receptor Dectin-1 was conducted. HEK-293 cells were transfected with mApple-tagged human Dectin-1a. Dectin-1 expression is sufficient to drive phagocytosis of C. albicans yeast cells by transfectants. The transfection conditions resulted in Dectin-1+ HEK-293, discriminated by positive mApple signal, and non-transfected cells, which were negative for mApple and served as an internal control to assess Dectin-1 dependence of binding and. phagocytosis. A flow cytometric assay of yeast cell binding to and internalization by HEK-293 transfectants was used. Yeasts were labeled with the pH-sensitive dye CypHer 5, which increases dramatically in emission intensity after internalization within acidic phagosomal compartments. The Cypher 5 signal was used to measure binding and internalization of yeast. Flow cytometry data were gated on HEK-293 cell-containing events for analysis, as defined by high side scatter signal, which was significantly larger than free yeast. Yeast bound to HEK-293 cells registered a low Cypher 5 signal. If yeasts were internalized, the CypHer 5 signal was much higher. The percent of HEK-293 cells with yeast bound (for mApple-Dectin-1+ and mApple-Dectin-1− cells) was determined by the percent of SSC-gated events having low or high CypHer 5 signal. The extent of phagocytosis was assessed by the median CypHer 5 fluorescence intensity within these populations.
For the results shown in
Despite their intrinsic resistance to cationic and oxidative stresses, C. albicans was highly susceptible to EO-OPE-DABCO, and to a lesser extent, PPE-DABCO. Biocidal activity of these compounds against C. albicans utilizes a dual mechanism combining light-independent cationic stress and light-dependent oxidative stress. Unlike other broad-spectrum antimicrobials, PEs exhibit low levels of in vitro toxicity against mammalian cells, making them intriguing candidates in numerous clinical applications.
Therefore, it is relevant to note that all clinical isolate strains exhibited significant amounts of killing during a 15 min exposure to EO-OPE-DABCO. Partial resistance of some clinical isolate strains may derive from adaptations of the pathogen to growth in the host, which may cause changes in cell wall structure and upregulation of mechanisms that permit growth under adverse conditions, such as leukocyte-derived ROS in the phagosomal environment.
While C. parapsilosis was found to be just as susceptible to EO-OPE-DABCO in the dark as C. albicans, C. glabrata displayed an inherent resistance. Candida spp. experience cationic stress as they interact with innate immune defenses. For example, cationic antimicrobial peptides, such as Histatnin-5, are deployed in host defense against Candida spp. and are thought to work by disrupting fungal plasma membrane integrity (REF). C. glabrata is noted for its resistance to killing by cationic antimicrobial peptides relative to C. albicans and other Candida spp. pathogens. Furthermore, C. albicans yeast cells display a modest stress response in the presence of heavy metal cations, activating 48 genes as a coping mechanism. Conversely, the cationic stress response in C. glabrata is more extensive, with over 100 genes being activated under similar circumstances. Cationic stresses imparted by heavy metals differs slightly from that of PEs; however, both are able to denature native protein conformation, and therefore may activate similar genes as part of a cationic stress response among Candida yeast cells. EO-OPE-DABCO's decreased ability to kill C. glabrata resembles the results of a previous study, in which a 10 μg/mL concentration of the compound failed to kill 99% of S. cerevisiae yeast, even after an hour in the light. Although S. cerevisiae is benign, it is closely related to C. glabrata. C. glabrata is also known to have robust antioxidative defenses that allow it to survive in the phagosome, and may impact its ability to resist oxidative killing by cationic phenylene ethynylenes.
It was found that PPE-DABCO strongly associates with soluble β-(1,3)-glucan (
Furthermore, PPE-DABCO displays immunostimulatory attributes, particularly in the light. This polymer was found to unmask the mannoprotein layer of C. albicans yeast cells in such a way that β-(1,3)-glucan could more easily be recognized and bound by pattern recognition receptor Dectin-1. PPE-DABCO binds to yeast cell walls (
The terms and expressions that have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the embodiments of the present invention. Thus, it should be understood that although the present invention has been specifically disclosed by specific embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those of ordinary skill in the art, and that such modifications and variations are considered to be within the scope of embodiments of the present invention.
The following exemplary embodiments are provided, the numbering of which is not to be construed as designating levels of importance:
Embodiment 1 provides a method of treating a fungal infection, the method comprising: contacting a fungus comprising a β-glucan that is at least partially masked from immune system detection with a therapeutically effective amount of a compound that at least partially unmasks the β-glucan to increase immunogenicity of the fungus.
Embodiment 2 provides the method of Embodiment 1, wherein the β-glucan comprises β-(1,3;1,6)-glucan.
Embodiment 3 provides the method of any one of Embodiments 1-2, wherein the fungus is a Candida species fungus, an Aspergillus species fungus, or a Histoplasma species fungus.
Embodiment 4 provides the method of any one of Embodiments 1-3, wherien the fungus is Candida albicans, Candida glabrata, Candida parapsilosis, or a combination thereof.
Embodiment 5 provides the method of any one of Embodiments 1-4, wherein the fungus comprises a biofilm.
Embodiment 6 provides the method of any one of Embodiments 1-5, wherein the fungus is in or on an indwelling medical device.
Embodiment 7 provides the method of any one of Embodiments 1-6, wherein the fungus is in or on a catheter.
Embodiment 8 provides the method of any one of Embodiments 1-7, wherein the compound is a polycationic conjugated aromatic system.
Embodiment 9 provides the method of any one of Embodiments 1-8, wherein the compound comprises a unit having the structure:
wherein
A1 is chosen from a bond.
A2 is chosen from a bond,
R1 is chosen from −H and. C1, wherein the compound comprises at least one C1,
at each occurrence, C1 is independently chosen from -L-D and —O-L-D,
at each occurrence, L is independently (C1-C10)hydrocarbylene,
at each occurrence, D is independently chosen from —N((C1-C5)alkyl)3 and a (C1-C10)alkyl-substituted cationic nitrogen-containing (C1-C5)heterocycle, and
n is about 1 to about 100,000.
Embodiment 10 provides the method of Embodiment 9, wherein the compound has the structure:
wherein
R2 is chosen from a bond and
R3 is chosen from a bond and
and
at each occurrence, T is independently chosen from —H, C(O)—O—(C1-C10)alkyl, -phenyl, and —R1.
Embodiment 11 provides the method of any one of Embodiments 9-10, wherein at each occurrence, L is independently chosen from methylene, ethylene, propylene, butylene, pentylene, and heptylene.
Embodiment 12 provides the method of any one of Embodiments 9-11, wherein the variable A1 is chosen from a bond,
Embodiment 13 provides the method of any one of Embodiments 9-12, wherein at each occurrence, D is independently chosen from —N(CH3)3,
Embodiment 14 provides the method of any one of Embodiments 9-13, wherein at each occurrence, D is independently chosen from —N(CH3)3,
Embodiment 15 provides the method of any one of Embodiments 9-14, wherein C1 is chosen from:
Embodiment 16 provides the method of any one of Embodiments 9-15, wherein at each occurrence, T is independently chosen from —H, C(O)—O-ethyl, -phenyl, and —R1.
Embodiment 17 provides the method of any one of Embodiments 1-16, wherein the compound is:
Embodiment 18 provides the method of any one of Embodiments 1-17, wherein the compound is a polymer comprising the repeating unit:
Embodiment 19 provides the method of Embodiment 18, wherein the polymer is a homopolymer.
Embodiment 20 provides the method of any one of Embodiments 1-19, wherein the compound is a polymer comprising the repeating unit:
Embodiment 21 provides the method of Embodiment 20, wherein the polymer is a homopolymer.
Embodiment 22 provides the method of any one of Embodiments 1-21, wherein the compound is a polymer comprising the repeating group:
Embodiment 23 provides the method of Embodiment 22, wherein the polymer is a homopolymer.
Embodiment 24 provides the method of any one of Embodiments 1-23, wherein compound is a polymer comprising the repeating group:
Embodiment 25 provides the method of Embodiment 24, wherein the polymer is a homopolymer.
Embodiment 26 provides the method of any one of Embodiments 1-25, wherein the compound is:
Embodiment 27 provides the method of any one of Embodiments 1-26, wherein the compound is:
Embodiment 28 provides the method of any one of Embodiments 1-27, wherein the compound is:
Embodiment 29 provides the method of any one of Embodiments 1-28, wherein the compound is:
Embodiment 30 provides the method of any one of Embodiments 1-29, wherein the compound is:
Embodiment 31 provides the method of any one of Embodiments 1-30, wherein the compound is:
Embodiment 32 provides the method of any one of Embodiments 1-31, wherein the compound is:
Embodiment 33 provides the method of any one of Embodiments 1-32, wherein the compound is:
Embodiment 34 provides e method of any one of Embodiments 1-33, wherein the compound is:
Embodiment 35 provides the method of any one of Embodiments 1-34, wherein the compound is:
Embodiment 36 provides a method of treating a fungal infection, the method comprising:
contacting a fungus comprising a β-glucan that is at least partially masked from immune system surveillance with a therapeutically effective amount of a compound that at least partially unmasks the β-glucan to increase immunogenicity of the fungus, wherein the compound comprises a unit having the structure:
wherein
A1 is chosen from a bond,
A2 is chosen from a bond,
R1 is chosen from —H and C1, wherein the compound comprises at least one C1,
at each occurrence, C1 is independently chosen from -L-D and —O-L-D,
at each occurrence, L is independently (C1-C10)hydrocarbylene,
at each occurrence, D is independently chosen from —N((C1-C5)alkyl)3 and a (C1-C10)alkyl-substituted cationic nitrogen-containing (C1-C5)heterocycle, and
n is about 1 to about 100,000.
Embodiment 37 provides a method of preventing or reducing a fungal infection on a substrate, the method comprising:
treating the substrate with a therapeutically effective amount of a compound so that contact between the treated surface and a fungus comprising β-glucan that is at least partially masked from immune system detection is effective to at least partially unmask the (β-glucan thereby increasing immunogenicity of the fungus and preventing or reducing fungal infection on the substrate from the fungus.
Embodiment 38 provides a method of preventing or reducing a fungal infection on or in a device, the method comprising:
treating the device with a therapeutically effective amount of a compound so that contact between the compound and a fungus comprising β-glucan that is at least partially masked from immune system detection is effective to at least partially unmask the β-glucan thereby increasing immunogenicity of the fungus and preventing or reducing fungal infection on or in the device from the fungus.
Embodiment 39 provides an antifungal compound comprising a unit having the structure:
wherein
A1 is chosen from a bond,
A2 is chosen from a bond.
R1 is chosen from —H and C1, wherein the compound comprises at least one C1,
at each occurrence, C1 is independently chosen from -L-D and —O-L-D,
at each occurrence, L is independently (C1-C10)hydrocarbylene,
at each occurrence, D is independently chosen from —N((C1-C5)alkyl)3 and a (C1-C10)alkyl-substituted cationic nitrogen-containing (C1-C5)heterocycle, and
n is about 1 to about 100,000,
wherein contact between a therapeutically effective amount of the compound and a fungus comprising β-glucan that is at least partially masked from immune system detection is effective to at least partially unmask the β-glucan thereby increasing immunogenicity of the fungus.
Embodiment 40 provides the method of Embodiment 39, wherein the compound has the structure:
wherein
R2 is chosen from a bond and
R3 is chosen from a bond and
at each occurrence, T is independently chosen from —H, C(O)—O—(C1-C10)alkyl, -phenyl, and —R1.
Embodiment 41 provides the method or compound of any one or any combination of Embodiments 1-40 optionally configured such that all elements or options recited are available to use or select from.
This application claims the benefit of priority to U.S. Provisional Patent Application Ser. No. 62/454,355 entitled “TREATMENT AND PREVENTION OF FUNGAL INFECTIONS,” filed Feb. 3, 2017, the disclosure of which is incorporated herein in its entirety by reference.
This invention was made with Government support under grant number 1R01AI116894-01A1 awarded by the National Institute of Allergy and Infectious Disease. The U.S. Government has certain rights in this invention.
| Number | Date | Country | |
|---|---|---|---|
| 62454355 | Feb 2017 | US |
